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Malaria in prehistoric sardinia (italy) :
b an examination of skeletal remains from the middle bronze age
h [electronic resource] /
by Teddi Setzer.
[Tampa, Fla] :
University of South Florida,
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Dissertation (PHD)--University of South Florida, 2010.
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ABSTRACT: Sardinia was an island with a history of a malarious environment until eradication efforts were conducted from 1946 to 1950. While historic documents suggest the disease was introduced from North Africa around 500 BC, no study has been conducted to test for the presence of malaria in prehistoric native populations, such as the Nuragic people of the Bronze Age. However, it has been suggested that aspects of the Nuragic culture, for example the stone structures found throughout the island, are adaptations to a malarious environment. The purpose of this dissertation is to test the hypothesis that malaria was present in prehistoric Sardinia. In addition, the value of applying anthropology, pertaining specifically to prehistoric investigations, to understand and combat malaria is supported. To test for the presence of malaria, multiple lines of evidence were used to analyze human skeletal remains from a Middle Bronze Age tomb. Because malaria does not result in a specific pattern of bony responses that can be identified through a gross analysis of the remains, additional lines of evidence were used. These included an osteological analysis for the possible presence of conditions related to malaria (e.g., inherited hemolytic anemias) and the collection of bone samples to test for ancient malaria DNA, Plasmodium falciparum histidine-rich protein II, and the malarial pigment hemozoin. In addition, a review of the literature pertaining to the ecology and history of Sardinia were used with archaeological data to evaluate if it was possible the malaria parasite was affecting humans on the island during prehistory. While it was interpreted that conditions were favorable for malaria to infect individuals during this time, and possible cultural adaptations were noted, no conclusive evidence was found by analyzing skeletal remains. More work is needed to diagnose malaria better in human remains and understand the health of populations in Sardinia during the Bronze Age. Considering the coevolution of malaria parasites, humans, and mosquitoes is a necessary step in developing methods to combat malaria as the parasite and disease vector become more resistant to medicine and insecticides. In particular, applying anthropological methods and theories shows promise for fighting this disease.
Advisor: Robert H. Tykot, Ph.D.
t USF Electronic Theses and Dissertations.
Malaria in Prehistoric Sardinia (Italy): An Examination of Skeletal Remains from the Middle Bronze Age by Teddi J. Setzer A dissertation submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy Department of Anthropology College of Arts and Sciences University of South Florida Major Professor: Robert H. Tykot, Ph.D. Erin Kimmerle, Ph.D. David Himmelgreen, Ph.D. Nancy Marie White, Ph.D. Lisa Kahn, Ph.D. Steven Reader, Ph.D. Date of Approval: July 1, 2010 Keywords: bioarchaeology, hemozoin, Medite rranean, porotic hyperostosis, thalassemia Copyright 2010, Teddi J. Setzer
Dedication To the people of Sardinia
i Table of Contents List of Tables iv List of Figures v Abstract xi Preface xiii Chapter 1: Introduction 1 1.1 The Importance of Using Social Science to Understand Malaria: Applied Anthropology 3 1.2 Sardinia, Malaria, and Archaeology 6 Chapter 2: Malaria 10 2.1 Morbidity, Mortality, and Costs 10 2.2 Types and Vectors 12 2.3 Disease Process and Symptoms 13 2.4 Acute and Chronic Malaria 16 2.5 Malaria and Other Diseases 18 2.6 Biological Adaptations 19 2.7 Cultural Adaptations 22 2.8 History and Debates 25 2.9 Chapter Summary 32 Chapter 3: Paleopathology, Archaeology, and Malaria 34 3.1 Bone 34 3.2 Analyzing Skeletal Remains in Populations Affected by Malaria 40 3.2.1 Porotic Hyperostosis, Cribra Orbitalia, and Anemia 42 3.2.2 Skeletal Indicators for Thalassemia 51 3.2.3 Ancient DNA 54 3.2.4 Immunology 58 3.2.5 The Biomarker Hemozoin 60 3.3 Chapter Summary 61 Chapter 4: Sardinia 63 4.1 Geography, Climate, and Environment 63 4.2 Humans and Sardinia 71 4.2.1 Upper Paleolithic and Mesolithic 72 4.2.3 Early Neolithic 73
ii 4.2.4 Middle Neolithic 74 4.2.5 Late Neolithic 76 4.2.6 Chalcolithic 81 4.2.7 The Bronze Age 83 4.2.8 The Nuragic Periods during the Iron Age 94 4.4 Malaria and Inherited Hemo lytic Anemias in Sardinia 97 4.5 Chapter Summary 107 Chapter 5: The Population, Materials, and Methods 109 5.1 Tomb B from Serra e Sa Caudeba 110 5.2 State of the Previous Analyses on the Remains from Serra e Sa Caudeba and Limitations of the Data 115 5.3 Methods of Data Collection and Analysis Used in this Research 119 5.3.1 Skeletal Inventories and Paleopathological Analysis of the Individuals 120 5.3.2 Paleopathological Analysis of Crania 122 5.3.3 Paleopathological Anal ysis of Other Remains 123 5.4 Collection of Bone Samples for aDNA, Pf HRP II, and Hemozoin Analysis 125 5.5 Preparation of Bone Samples for aDNA, Pf HRP II, and Hemozoin Testing 126 Chapter 6: Skeletal Data and Interpretation 129 6.1 Mortuary Treatment of the Remains and Taphonomy 129 6.2 The Cist Burials 130 6.3 Results of the Analysis of Individual Skeletons 131 6.4 Results of the Analysis of Cranial Bones 133 6.5 Overall Osteology Results Includi ng Postcranial Bone s, Individuals, and Crania 138 6.6 Results of Malarial aDNA, Pf HRP II, and Hemozoin Analysis from Serra e Sa Caudeba Tomb B 139 6.7 Conclusions 139 Chapter 7: Discussion 141 7.1 The Biocultural Approach in Anthropological Research 141 7.2 Ecological and Archaeological Evidence of Malaria in Nuragic Populations 144 7.2.1 Is there evidence of cultura l adaptations to malaria in Bronze Age Sardinia? 144 7 .2.2 Did sufficient changes to the ecosystem occur to support the disease vector before or during the Bronze Age? 146 7.2.3 Did subsistence patterns of Nuragic populations result in land modification conducive to malaria? 147 7.2.4 Is there evidence of contact with other populations that may have been malarious before or during the Bronze Age? 147
iii 7 .2.5 Can ethnographic analogies be made to Nuragic populations? 149 7.3 Discussion of Osteological Research 150 7.4 Discussion of Skeletal Evidence from Serra e Sa Caudeba Tomb B 156 7.4.1 Mortuary Treatment of the Remains and Taphonomy 156 7.4.2 Discussion of the Cist Burial 158 7.4.3 Differential Diagnosis from Individual Skeletons 159 7.4.4 Results and Interpretation of th e Analysis of Cranial Bones 160 7.4.5 Overall Osteology Results Including Postcranial Bones, Individuals, and Crania 168 7.5 Discussion of Malarial aDNA, Pf HRP II, and Hemozoin Analysis from Serra e Sa Caudeba Tomb B 171 7.6 Chapter Summary 172 Chapter 8: Conclusions 176 8.1 Major Findings 176 8.2 Strengths and Limitations 178 8.3 Contributions and Implications 180 8.4 Directions for Future Research 181 8.5 Summary and Conclusions 184 References Cited 186 Appendices 230 Appendix A: Artifacts fro m Serra e Sa Caudeba 231 Appendix B: Data Collection Protocol 236 Appendix C: Raw Data 251 Appendix D: Photographs of Pathologies Observed on the Individual Skeletons 285 Appendix E: Photographs of Examples of Common Pathologies in this Population 298 Appendix F: Photographs of Examples of Unusual Pathologies in this Population 302 Appendix G: Photographs of Bones Used for Sampling for Ancient DNA, Pf HRP II, and Hemozoin Analysis 314 About the Author End Page
iv List of Tables Table 3.1. Factors affecti ng iron status (after Stuart-Macadam 1998:Table 4.5) 47 Table 4.1. The chronology of Sardinia (after Tykot 1994:129) 71 Table 5.1. MNI of individuals from Tomb B 116 Table 5.2. Table5.2: Inventory of el ements from Tomb B by weight 118 Table 5.3. Bone samples collected for ancient DNA, Pf HRP II, and hemozoin analysis 125 Table 5.4. Bone samples from tomb B prepared for ancient DNA, Pf HRP II, and hemozoin testing 127 Table 6.1. Pathological conditions obser ved on the individual skeletons 132 Table 6.2. The occurrence and distribution of porotic hyperostosis and cribra orbitalia observed in the sample of crania analyzed from this population 134 Table 7.1. Recommended Dietary Allowan ces and Adequate Intakes for iron and vitamins A, B6, B9, B12, and C per day (Food and Nutrition Board 2006) 162 Table 7.2. Nutrient values for foods found in the archaeological record at Neolithic and Bronze-Age sites in Sardinia. All values are per 100 g, with the exception of olive oil, which is per tablespoon. From http://www.nal.usda.gov/fnic/foodcomp/search/ 163 Table 7.3. The number of observations of each pathological condition observed, and the association of these conditions with thalassemia and tuberculosis (after Ortner and Putschar 2003) 169 Table C.1. Pathologies Observed on Individual Skeletons 265 Table C.2. Raw Data from Cranial Elements 267 Table C.3. Raw Data from Individual Elements 276
v List of Figures Figure 2.1. The malaria life cycle and in fection process (courtesy of CDC/ Alexander J. da Silva, Ph.D./Me lanie Moser from the Centers for Disease Control Public Health Image Library) 13 Figure 2.2. Map identifying th e location of the thalasse mia belt. Sardinia is identified with an arrow (www.outline-world-map.com). 21 Figure 3.1. Histological schematic of co rtical and trabecular bone (U. S. National Institutes of Health 2010) 35 Figure 3.2. An interpretation of bone grow th from a cartilage model (U. S. National Institutes of Health 2010) 38 Figure 3.3. An example of porotic hyperostosis on a parietal bone fragment from Serra e Sa Caudeba, cranium e2 (photo by author 2008) 43 Figure 3.4. An example of cribra orbitalia on a frontal bone fragment from Serra e Sa Caudeba, cranium 3 (photo by author 2008) 43 Figure 4.1. The location of Sardinia and Corsica in relation to Sicily and the mainland (image by the Sea WiFS Project, NASA/Goddard Space Flight Center, and ORBIMAGE 2006) 64 Figure 4.2. Map of Sardinia with locati ons discussed identified (image by Jeff Schmaltz, MODIS Rapid Response Team, NASA/GSFC 2003) 65 Figure 4.3. Exterior (above,) and interi or (below) of the hypogea at Anghelu Ruiu, 3rd millennium BC (images courtesy of Stephanie Del Rosario) 78 Figure 4.4. Monte dAccoddi (image courtesy of Robert Tykot) 80 Figure 4.5. The Su Nuraxi Barumini complex (image courtesy of Robert Tykot) 84 Figure 4.6. Nuraghe Losa is an example of a proto-nuraghe. It is located in central-western Sardinia (ima ge courtesy of Robert Tykot) 88
vi Figure 4.7. Well temple at Santa Cristina (photo by author 2005) 90 Figure 4.8. S Ena e Thomes, a giants tomb, located in Northern Sardinia (image courtesy of Robert Tykot) 92 Figure 4.9. Spread of malaria as inte rpreted by historic reconstruction (after Sallares et al. 2004; image by the SeaWiFS Project, NASA/Goddard Space Flight Ce nter, and ORBIMAGE 2006) 99 Figure 5.1. Serra e Sa Caudeba is locat ed on the Campidano Plain (image by Jeff Schmaltz, MODIS Rapid Response Team, NASA/GSFC 2003). Figure 5.2. The entrance of Tomb A is built into the knoll (photo by author 2008). 111 Figure 5.3. Tomb B was di scovered during the constr uction of a road. Only half of the tomb remains intact (photo by author 2008). 112 Figure 5.4. Tomb B during the excavation (i mage courtesy of Alessandro Usai) 113 Figure 5.5. The Civico Museo Archeologico in Villanovaforru (photo by author 2008) 114 Figure 5.6. Bar graph of the MNI from Tomb B (from the discrete categories provided) 117 Figure 5.7. Example of the condition of the cr anial remains from Serra e Sa Caudeba (photo by author 2008) 122 Figure 5.8. Remains from Serra e Sa Caudeba Tomb B with contextual information (photo by author 2008) 124 Figure 5.9. Processing of the samples with EDTA to maximize the chemical reaction before further te sting (photo by author 2009) 128 Figure 6.1. Taphonomic observations from Serra e Sa Caudeba (from upper left clockwise: insect ac tivity, post-excavation damage, and evidence of rodent gnawi ng, photos by author 2008) 130 Figure 6.2. Crania observed with and wit hout porotic hyperostos is and cribra orbitalia 137 Figure 6.3. Distribution of cas es of porotic hyperostosis and cribra orbitalia by bone (the number of frontal bone observations is the total for both conditions) 137
vii Figure 6.4. The frequency of the pathol ogical categories observed at Serra e Sa Caudeba Tomb B (after Buikstra and Ubelaker 1994:112) 138 Figure 7.1. Distribution of obsidian from Monte Arci during the Neolithic (reprinted with permission from Tykot 2004) 148 Figure A.1. Incised truncated terracotta cup, or tazza from Tomb A 232 Figure A.2. Clay beads from Tomb A 232 Figure A.3. Bronze dagger blade from Tomb A 233 Figure A.4. Hemispheric terracotta cup from Tomb A 233 Figure A.5. Terracotta globul ar urn from Tomb B 234 Figure A.6. Clay beads from Tomb B 234 Figure A.7. Terracotta globul ar urn from Tomb B 235 Figure A.8. Terracotta sherds and urn lid (or plate) from Tomb B 235 Figure D.1. Photograph 6: myositis ossificans of the left femur of individual b 286 Figure D.2. Photograph 7: spicule/myositi s ossificans of the left fibula of individual b 286 Figure D.3. Photograph 8: periosto sis of the rib of individual b 287 Figure D.4. Photograph 9: gradual cha nge in body height, bone loss, and osteophytes (barely discernable) of a thoracic vertebrae of individual b 287 Figure D.5. Photograph 10: Schmorl's node of a vertebral fragment of individual b 288 Figure D.6. Photograph 11: osteophytes of a vertebral fragment of individual b 288 Figure D.7. Photograph 42: syndesmophytes of a vertebral body of individual c 289 Figure D.8. Photograph 43: osteophytes of a vertebral body of individual c 289 Figure D.9. Photograph 44: gradual change in body height and osteophytes of a vertebral body of individual c 290
viii Figure D.10. Photograph 45: osteophytes of a vertebral body fragment of individual c 290 Figure D.11. Photograph 46: fracture of the left clav icle of individual c 291 Figure D.12. Photograph 47: fracture of the right fibula of individual c 291 Figure D.13. Photograph 48: arthritis of the right humerus of individual c 292 Figure D.14. Photograph 49: arthritis of the right humerus of individual c 292 Figure D.15. Photograph 51: multifocal ly tic lesions of the right ilium of individual c 293 Figure D.16 Photograph 32: dislocati on of the right glenoid fossa of individual eI 294 Figure D.17 Photograph 167: reactive woven bone, porosity, and eburnation of the proximal tibiae of individual 6 (bilateral) 294 Figure D.18 Photograph 168: myositis ossi ficans of a fibula of individual 6 295 Figure D.19 Photograph 169: Schmorl's node of a thoracic vertebrae of individual 6 295 Figure D.20 Photograph 170: Schmorl' s node, osteophytes, and spicule formation were observed on a thor acic vertebra of individual 6 296 Figure D.21 Photograph 171: oste oarthritis of the right hum erus of individual 6 296 Figure D.22 Photograph 172: osteoarthritis of the left femoral head of individual 6 297 Figure D.23 Photograph 173: osteoarthritis of the left femoral head of individual 6 297 Figure E.1. Endocranial lesion (cra nial fragment from W2-W3) 299 Figure E.2. Porotic hyperostosis (parietal from cranium e2) 299 Figure E.3. Cribra orbi talia (cranium c2) 300 Figure E.4. Angling of vertebral body (q4-180) 300 Figure E.5. Lytic lesion of vertebral body (q4-180) 301
ix Figure E.6. Arthritis of femur 301 Figure F.1. Hypoplasia of mastoid proce ss (presented as bilateral in mp1; right side pictured) 303 Figure F.2. Meningeal reaction with granular impressions (cranium 3) 303 Figure F.3. Depression fract ure of parietal bone ( cranium taglio nella terra ) 304 Figure F.4. Cloaca (tibia) 304 Figure F.5. Fracture (juvenile humerus) 305 Figure F.6. Lesion sternum (q1-180) 305 Figure F.7. Necrosis of the femur (q13-140) 306 Figure F.8. Ankylosis of cervical vertebrae ( 1 scatola ) 306 Figure F.9. Lesions of the sternum 307 Figure F.10. Arthritis (acetabulum) 307 Figure F.11. Arthritis ph alange (q12-q14 -140-160) 308 Figure F.12. Arthritis (ulna) 308 Figure F.13. Lytic lesion navicular (q9) 309 Figure F.14. Arthritis ulna (q15-130) 309 Figure F.15. Lesion calcaneus (q0-185) 310 Figure F.16. Lesion ulna (q13-140) 310. Figure F.17. Myositis ossificans of an ulna, possibly from a fracture (fragment q3-180) 311 Figure F.18. Associated bony reaction in the adjacent radius (fragment q3-180) 311 Figure F.19. Porosity of vertebra ( 1 scatola ) 312 Figure F.20. Porosity of vertebra ( 1 scatola ) 312 Figure F.21. Hyperostosis of long bone fragment (w4-x4) 313
x Figure G.1. Sample 1: right humerus fragment, infant ( bambini e adolescenti ) 315 Figure G.2. Sample 2: left humerus, infant ( bambini e adolescenti ) 315 Figure G.3. Sample 3: long bone fragment, infant ( bambini e adolescenti ) 316 Figure G.4. Sample 4. Long bone fragment, infant ( bambini e adolescenti ) 316 Figure G.5. Sample 5: left ulna fragment, adolescent ( sepoltura de fronte alla tomba B ) 317 Figure G.6. Sample 6: left humerus fragment, adolescent (Q1 ; quota 180/240 tomba B ) 317 Figure G.7. Sample 7: unsided hum erus fragment, adolescent (Q16 ; quota 130 tomba B ) 318 Figure G.8. Sample 8: unsided humerus fragment, adult (Q1; quota 180 tomba B ) 318 Figure G.9. Sample 9: right humerus, adult (W4; ex primo taglio tomba B ) 319 Figure G.10. Sample 10: right humerus, adult (Quota 177 tomba B) 319
xi Malaria in Prehistoric Sardinia (Italy): An Examination of Skeletal Remains from the Middle Bronze Age Teddi J. Setzer Abstract Sardinia was an island with a history of a malarious environment until eradication efforts were conducted from 1946 to 1950. While historic documents suggest the disease was introduced from North Africa around 500 BC, no study has been conducted to test for the presence of malaria in prehistoric na tive populations, such as the Nuragic people of the Bronze Age. However, it has been s uggested that aspects of the Nuragic culture, for example the stone struct ures found throughout the island, are adaptations to a malarious environment. The purpose of this dissertation is to te st the hypothesis that malaria was present in prehistoric Sard inia. In addition, th e value of applying anthropology, pertaining speci fically to prehistoric inve stigations, to understand and combat malaria is supported. To test for the presence of malaria, mu ltiple lines of evidence were used to analyze human skeletal remains from a Mi ddle Bronze Age tomb. Because malaria does not result in a specifi c pattern of bony responses that can be identified through a gross analysis of the remains, additional lines of evidence were used. These included an osteological analysis for the possible pres ence of conditions related to malaria (e.g., inherited hemolytic anemias) and the collect ion of bone samples to test for ancient
xii malaria DNA, Plasmodium falciparum histidinerich protein II, and the malarial pigment hemozoin. In addition, a review of the litera ture pertaining to the ecology and history of Sardinia were used with archaeological data to evaluate if it was possible the malaria parasite was affecting humans on the island during prehistory. While it was interpreted that conditions were favorable for malaria to infect individuals during this time, and possible cultural adaptations were noted, no conclusive evidence was found by analyzing skeletal rema ins. More work is needed to diagnose malaria better in human remains and understa nd the health of populat ions in Sardinia during the Bronze Age. Considering the coevol ution of malaria para sites, humans, and mosquitoes is a necessary step in developing methods to combat malaria as the parasite and disease vector become more resistant to medicine and insecticides. In particular, applying anthropological methods and theories shows promise for fighting this disease.
xiii Preface In 2002, I made my first trip to Sard inia with Robert H. Tykot to conduct excavations and begin obsidian use-wear e xperiments at the volcanic complex of Monte Arci. This trip not only sparked my intere st in Sardinian archaeology, but it was the beginning of my academic relationship with the island. My masters thesis considered the use of Monte Arci obsidian in the Neolithic, and through subsequent projects, I examined its changing role through the Br onze Age. This interest led to more work in Sardinia. I was given the opportunity to work as sta ff for the University of South Floridas archaeological field schools in Sardinia in 2005 and 2006 with continued work at Monte Arci and the nuraghe, Nuracale. During this wo rk, I was introduced to Dott. Carlo Lugli, Dott. Mauro Perra, Dott. Alessando Usai, and Dott.ssa Ornella Fonzo. I am grateful to them for their never-ending assistance thr oughout my dissertation pr ocess. In addition, this research would not have been possible without the permission of the Soprintendenza per i Beni Archeologici della Sardegna, and Do tt. Fulva Lo Schiavo. I would also like to acknowledge the contributions of Dr. Luca Lai. I found the many conversations about Sardinian archaeology, including suggesting st udying the remains from Serra e Sa Caudeba, helpful to designing this researc h. Through his friendship, I was able to understand Sardinia and its people better. Many individuals provided training and consult for the osteological and ancient DNA portion of the research. In particular, I would like to thank Dr. David Himmelgreen and Dr. Erin Kimmerle for giving me ma ny learning opportunities in biological
xiv anthropology. Dr. Jane E. Buikstra and Dr Alicia Kay Wilbur provided me with a foundation in human osteology and an intere st in ancient DNA analysis. Additional assistance regarding bone coll ection was given by Dr. Abig ail Bouwman and Dr. Terry Brown from the Manchester In terdisciplinary Biocentre. The services, facilities, and related expenses for th e ancient DNA, protein, and hemozoin analysis were provided by Dr. Da vid Sullivan, Jr., Dr. John Pisciotta, and Dr. Abhai K. Tripathi of the Johns Hopkins Mala ria Research Institute at the Johns Hopkins Bloomberg School of Public Health. These porti ons of this research could not have been completed without their exper tise, collaboration, time, and patience. I am thankful to them for providing assistance with the preparation of the material and performing these analyses, which were integral to the mu ltiple-lines-of-evidence approach in this dissertation. In addition, I w ould like to acknowledge and th ank Robert D. Bowers for assisting in preparing the University of South Floridas Laboratory for Archaeological Science for the processing of these samples, and the Cell Biology, Microbiology and Molecular Biology (CMMB) Research Facilities for the use of materials and equipment needed to prepare the samples. I am also thankful to Dr. Robert H. Tykot for introducing me to Sardinian archaeology and providing years of mentori ng and countless opportunities throughout my graduate school career. I woul d like to thank Dr. Nancy Marie White for her years of advice and feedback, Dr. Steven Reader for he lping me to consider the role of geography in health-related issues, and Dr. Lisa Ka hn for her feedback and knowledge throughout this process. All of this has influenced my work.
xv My friends and family have also contri buted to this process through their support and encouragement. Many friends, family member s, and coworkers, too numerous to list completely, have cheered me on throughout this process. However, in particular, I would like to thank Stephen Carmody and Nicole Shelnut for their seemingly endless consideration and support. My mother and fa ther, Margaret and Theodore Setzer, were always encouraging. They provided me with an appreciation for the past, an inquisitive nature, and a desire to explor e. In addition, my sister, Ca thy DeFelice, always supported my interest in science. To them I am gr ateful beyond words. Fi nally, I would like to thank my son, Braden, for motivating me in a way no other could.
1 Chapter 1: Introduction This research tests the hypothesis that malaria wa s present during the Middle Bronze Age in Sardinia. This dissertation is one of an applied nature. That is, anthropological methods, theories, and perspe ctives, specifically fr om the subfields of archaeology and physical anth ropology, are used to addres s the presence of malaria on the Mediterranean island of Sardinia during prehistory. This information is used in conjunction with ethnographic research (B rown 1986; Inhorn and Brown 1990:93) to identify and understand possible biocultural adap tations to malaria that contributed to the success of populations on this island. In turn, the identificatio n and understanding of these biocultural adaptations and other anthr opological data can be used when developing new approaches to combat the transmissi on of malaria in c ontemporary populations. First, the role of applying social sciences, in particular anthropology, is presented to demonstrate the need for understa nding human behavior and host/pathogen coevolution to combat the disease effectiv ely. Current knowledge about malaria and its adaptive responses (biol ogical and cultural), the reaction of skeletal tissue to pathological and stressful conditions, and Sardinia itself (from preh istory through ethnographic evidence from modern populations) is presen ted. For each of these topics, conditional statements of what may be empirically observed in the archaeological record are generated. The hypothesis is test ed through deductive reasoning.
2 In addition, this research includes an osteological examination. Working with human skeletal remains involves many ethi cal considerations (White 2000:330-332). For example, as with any archaeological remains and sites, bones contain information about questions that cannot be addr essed yet. Therefore, it is desirable to keep museum collections intact for future research and technologies. Since the excavation and handling of human remains has symbolic importance to many people, the remains used in this research were treated with care and respect. A minimal number of samples were taken to address the research questions, and multiple tests were conducted on the same sample. In addition, the remains examined in this study were housed in the Civico Museo Archeologico in Villanovaforru, Sardinia, which is located near the archaeological site and is the home of individuals with the most likely direct lineal affinity. The population of this town was informed of the resear ch through meetings with the mayor and publication of an article in a local newspaper. Permission to study the remains was obtained from the Soprintendenza per i Beni Ar cheologici di Caglia ri. In addition, a copy of this dissertation will be made availa ble to the museum and the office of the soprintendenza. In this research, bone tissue is analyz ed through the use of multiple lines of evidence (i.e., gross paleopat hological analysis, ancient malarial DNA testing, protein analysis, and hemozoin [malaria pigment] detection techniques) to study remains from Tomb B of the Middle Bronze Age (Nuragic) si te of Serra e Sa Caudeba. Overall, an examination of the archaeological and oste ological evidence is made to assess the likelihood that malaria was present on Sa rdinia during the Middle Bronze Age. The results of this research were inconclusive. Although the DNA test ing produced positive
3 results that could not be rep licated (false positives) and th e protein and hemozoin results were negative, the review of literature and the osteological data and archaeological evidence do not refute the hypothesis that malaria was present. However, these observations do not permit the null hypothesi s (malaria was not present during the Bronze Age) to be rejected, either. The obser vations and results of this research are interpreted and discussed within a biocu ltural framework. The significance of these findings and directions for future research are also presented. The challenges and additional considerations of the interpretation of these data, in particular those concerning bony responses, are discussed in Chapters 7 and 8. In addition, Nuragic people are often refe rred to as warrior pa storalists (Lazrus 1999; Rowland 2001). Although not related to th e testing of the hypothesis that is the subject of this dissertation, the limited am ount of trauma observed during this study is noteworthy. While it is possible that individuals who were in jured or killed during battles were interred elsewhere, no evidence of sepa rate burials for warriors has been discovered. The results of this analysis do not su pport the warrior-pasto ralist hypothesis. 1.1 The Importance of Using Social Scie nce to Understand Malaria: Applied Anthropology Each year, 3.3 billion peopl e are at risk for malaria, a disease affecting over 250 million individuals and causing almost one m illion deaths annually, most of which occur in children of Sub-Saharan Africa (W orld Health Organization 2009a, 2009b). The severity of this disease has received atte ntion from many parties, including the World Health Organization and the Centers for Di sease Control and Prevention, whose primary
4 foci consist of examining the parasite a nd its transmission, generating epidemiological data, and making recommendations regarding the prevention of ma laria. In addition, several non-profit groups, such as The Bill and Melinda Gates Foundation, Malarial No More, the Clinton Global Initiative, and the Ma laria Foundation International, to name a few, have implemented programs to fight this disease. Examples of their approaches include the distribution of mosquito nets, in secticides, and medicine, as well as funding research for malarial vaccines and no n-resistant drugs, and providing community education. While ethnomedical and ecological k nowledge is used to combat malaria (see World Health Organization 2009a, 2009b), thes e programs overwhelmingly operate from the perspective of a biomedical paradi gm, relying on treating individuals and communities with the administration of drugs a nd the application of pesticides. However, this has led to the emergence of drug and pe sticide resistant strain s of malaria (Baleta 2009), which in turn requires more research in drug development. Program planners are finding many control measures ineffective or too expensive, and they agree new methods of prevention must be developed (Oakes 1991). The Committee for the Study on Malaria Prevention and Control (Oakes 1991:258; Williams et al. 2002) recommends us ing applied social science to address problems associated with malaria. In partic ular, using anthropological approaches in malaria research is valuable because malari a acts upon behavioral ch aracteristics as well as having a biological impact. Fu rthermore, the distribution and transmission of malaria is affected by human actions, such as envir onmental modifications and trade (Stratton et al. 2008). Inhorn and Brown (1990:109) noted diseases, such as malari a, are the leading cause of death in societies studied traditiona lly by anthropologists. Anthropologists are
5 helpful in the process of disease management because they understand that cultures form social constructions of various aspects of di sease, such as causation and treatment, while the biomedical approaches are limited by their own concepts of the disease and treatment processes. Also, while malaria causes human su ffering, there is ultimately an aspect of this suffering involving political and socio economic inequality (i.e., political economy of health); although, some research has indi cated no correlation exists between socioeconomic status and malaria in Afri ca (Koram et al. 1995; Luckner et al. 1998; Najera 1994). Even if that is the case, the nature of the biomedicinal institution maintains social inequalities (Inhorn and Brown 1990:109) To deliver effective treatments and programs to improve the conditions caused by infectious disease, anthropologists must be equipped to work within the biomed ical paradigm (Inhorn and Brown 1990:109; Najera 1999). While the theories and methods of the social sciences have played little or no role in the development of various malaria control programs, they are considered to be useful when identifying local beliefs about mala ria and behaviors that can reduce its transmission. In particular, medical anth ropology can provide a unique perspective because of the holistic nature of the disc ipline. When examining health and human behavior, medical anthropologi sts understand the complexity of a culture must be considered in its larger context (e.g., historic economic, geographic, biologic, social, and political), rather than in isolation (Williams et al. 2002:251) For instance, epidemics of malaria can be managed by understanding how behaviors, such as deforestation or population movement, affect the viability and transmission of the parasite (Oakes et al. 1991:258).
6 By studying malaria in preh istoric and histor ic populations, th e antiquity and evolution of the parasite and the balanced polymorphisms th at provide protection from the disease can be established. Furthermore, knowing if malaria was present in a population provides an additional context for understanding a culture (Inhorn and Brown 1990:93). Researching the role of parasites in archaeological remains can result in a better interpretation of human behavior, including diet, trad e, subsistence, health and disease, human-parasite ecology, biocultura l adaptation, and issues involving contact (Reinhard 1992). 1.2 Sardinia, Malaria, and Archaeology Sardinia was known historically as one of the most malarious regions in the Mediterranean until an experiment to eradi cate malaria was carried out on the island from 1946 to 1950 by the Ente Regionale per la Lotta Anti-Anofe lica in Sardegna (ERLAAS), the United Nations Relief and Rehabilitati on Administration (UNRRA), the Economic Commission for Africa (ECA), and the Rock efeller Foundation (Bruce-Chwatt and de Zulueta 1980:102-105). High rates of biologi cally-adaptive conditions for malariaendemic environments are observed on this island. For example, thalassemia, an inherited hemolytic anemia offering protection from malaria, is found on Sardinia. Another adaptive condition, glucose-6-phosphate dehydro genase (G6PD) deficiency, is also present on Sardinia (Sanna et al. 1997:294-295). It is thought, through the anal ysis of historical Roman and Greek documents, that malaria was introduced on the island at about 500 BC from North Africa. However, this interpretation is formulated from unteste d hypotheses and does not fully incorporate the
7 information we now have from archaeological, ecological, genetic and entomological research. Therefore, it is not known if malari a (and if so, what sp ecies) was afflicting the people of Sardinia before 500 BC. Scholars ha ve contemplated this question during their research with historic populations adapta tions to malaria (Brown 1984) and ancient deoxyribonucleic acid (aDNA) studies (Sal lares and Gomzi 2001; Soren 2003). They have offered suggestions on how to addr ess identifying malari a in prehistoric populations, and they have formed hypotheses regarding possible cultu ral adaptations to malaria in prehistoric Sardin ia. For example, Brown (1984) suggests examining skeletal remains in a similar manner to that of phys ical anthropologist J. Lawrence Angel (1966, 1972), who examined skeletal, archaeological, demographic, and environmental data to conclude malaria was present in the Ae gean during the Neolithic. Many conditions, however, can result in skeletal lesions similar to those Angel observed. With advancements in the detection of mala ria aDNA (Sallares and Gomzi 2001), multiple lines of evidence can now be used when addressing this topic. Investigating the presence of malaria in the archaeol ogical record relies on a primarily processual approach, which has ju st started to gain acceptance in Sardinia during the last two decades. Because the island was subjected to outside forces throughout history, Sardinian ar chaeologists often make inte rpretations derived from invasion-resistance models, which result in a Sardinian/anti-colonialist bias. For example, the interior regions of Sardinia are often inte rpreted as being resist ant to change and the coastal regions are thought to have been subjected to outside influence (Dyson and Rowland 2007:13). Moreover, because of economic development on the island, thousands of sites on Sardinia have been ex cavated with little done beyond salvaging the
8 archaeological remains. While these salvaged archaeological remain s are numerous and provide many opportunities for research, the short timeline associated with recovery projects associated with cons truction can restrict the resear ch questions posed and impact the recovery and curation tec hniques. For example, in the case of the human remains studied here, the site was identified during the construction of a road. It was necessary for archaeologists to excavate quickly, thus a ffecting decisions regarding the recovery approaches, the recording of the site and, in turn, the ca taloging and curation of the human remains and artifacts. In addition, sin ce some of the materials were excavated many years ago, they were not collected w ith the influence of newer analytical techniques. Fore example, aDNA analysis wa s not considered at the time, and remains were not handled with a protocol that prevented contamination with modern DNA. The methods used in this research require the analysis of human skeletal remains. Prior osteological studies of populations from the Neolithic through Medieval periods in the Central Mediterranean can be classified into the following categories: pathological case studies (Brasili et al. 1997, 2002; Canci et al .1992; Capasso et al. 1996; Fornaciari et al. 1985-1986; German and Capitanio 1985-1986; Kritscher 1989; Mallegni et al. 2000; Passarello and Spoliti 1982-1983; Repetto and Canci 1987; Repe tto et al. 1988; Soren et al. 1995; Sperduti and Manzi 1990); descrip tive accounts of skeletal remains (Atzeni 1967; Bedini 1988; Di Salvo and Tusa 1990; Drusini et al. 2000; Formicola 1986-1987; German 1982; Giusberti et al. 1997; Mall egni, and Fornaciari 1979-1980; Marchi and Borgognini 2002; Minozzi et al. 1994; Novak and Knsel 1997; Passarello 1984-1985; Robb and Mallegni 1994; Schwidetzky and Ramaswamy 1980; Sonego and Scarsini 1994); nutrition/dietary analys es (Cook 2000; Lai 2008; Papath anasiou et al. 2000 ); and
9 status/activity related stud ies (Becker 2000; Kirkpatrick Smith 2000; Robb 1994; Robb et al. 2001). Examples of research addressing malaria specifically include Angel (1966; 1967; 1972; 1978), with his work in the Easter n Mediterranean, and Sallares and Gomzi (2001) and Soren (2003), who analyzed rema ins from a Roman cemetery in Teverina, Italy. While this research tests the hypothesis that malaria was present in Sardinia during the Bronze Age, another goal of this study is to ga in a better understanding of the lives and behaviors of Nuragic populations who lived during this time. If Nuragic populations were faced with malaria, the material remains and behaviors are better understood. For example, research with liv ing populations has demonstrated many cultural adaptations to malaria are non-deliberate; that is, thes e behaviors are not perceived to be related to ma intaining health, yet they pres erve or enhance health (Dunn 1976; Wood 1979). Is it possibl e architectural features on the island provided protection from exposure to mosquitoes carrying malaria (see Flumene in Brotzu 1934; Sallares 2002)? Is an unusual cist burial with juvenile remains related to a malarial epidemic (Soren 2003)? Was this disease present onl y when settlement patterns became more nucleated? How do the results relate to subsis tence practices and diet ary data (Lai 2008)? What is the relationship between the results of this study and sociopolitical development in the Nuragic culture? How does this informa tion relate to the evidence of social and economic interaction?
10 Chapter 2: Malaria To examine a disease in prehistory, one must have an understanding of the condition in history. By know ing the impacts malaria has on living populations, it becomes evident that research providing a new pe rspective of the pathog en is valuable. In the case of malaria, being familiar with the di fferent species of malarial parasites, the disease vectors, and its interaction with other infections is integral to the process. This, with the knowledge of biological and cultura l adaptations, can give archaeologists the information needed to understand health and human behavior in history and prehistory better. Much of the research (e.g., Warre ll and Gilles 2002) concerning malaria has focused upon the clinical manifestations and practices needed to asse ss and treat living populations; however, with advances in gene tic research, biological scientists have started to delve into questions concerning the timeframe of th e origin and th e evolution of malarial parasites. This basic understandi ng of malaria and its impact upon a population is essential for identifying conditions that ca n be observed in the ar chaeological record. 2.1 Morbidity, Mortality, and Costs Estimating the number of deaths occurring annually from malaria infections is difficult. Many areas where malaria is endemic lack formal health care systems and the ability to study the disease. Often, in cases where the disease is monitored, facilities capable of diagnosing parasitic infections ma y not be available, or the cost of the
11 associated laboratory work to diagnose the disease may be prohibitive. Thus, researchers rely often upon making a diagnosis based sole ly from symptoms or accounts of previous infections, rather than the verification of parasitemia (parasites in the blood). Unfortunately, there are many diseases with symptoms similar to those of malaria in these areas. This causes misdiagnosis to be a co ncern when considering the impact of this disease (Breman 2001:1). Even though researchers are faced with th ese challenges, they are able to make estimates regarding the number of infections and deaths resulting from malaria. While many efforts have been made to control the disease, it is estimated 700,000 to 2.7 million people still die from malaria each year, w ith over 75% of these deaths occurring in African children (Snow et al. 1999). In addition to the deaths associated with malaria, African children, especially those who are unde r the age of five, are susceptible to many malaria infections per year. Reports have b een made of four to nine febrile episodes occurring typically per year in an infected individual, re sulting in a total of 400,000 to 900,000 malaria-related fevers in African ch ildren under the age of five each year (Breman 2001:1,7). While malaria is not only a cause of great concern from a health standpoint, it also has social and economic impacts (see Breman 2001:1; Gallup and Sachs 2001; Holding and Snow 2001; Sachs and Malaney 2002:680) These include effects upon population growth, fertility, premature mortality, medical costs, trade, tourism, and worker productivity. Sachs and Malaney (2002:681) not e the global distribution of malaria is strongly correlated to areas with high poverty rates, namely th e tropics. However, it is not known if this is a causa l relationship in these regions. Fu rthermore, while the disease has
12 been eradicated in some areas, such as the United States and Eur ope, behaviors such as international tourism, immigration, and intravenous drug use have resulted in a resurgence of malaria in some areas (Caste lli et al. 1993; Oakes 1991), creating further challenges and concerns. 2.2 Types and Vectors Malaria is caused by a protozoan parasite ( Plasmodium sp. ) transmitted by mosquitoes of the Anopheles genus. Infection is not limite d to humans, and many species of this parasite impact re ptiles, birds, rodents, and non-human primates. While some species that infect these animals may cau se malaria in humans, there are four Plasmodium species most common in humans. These are P. vivax, P. falciparum P. malariae and P. ovale (in order of most to least infectious) (Mendis et al. 2001). P. vivax is the most widely distributed, and th e most common form where it occurs. P. falciparum is a major threat in Brazil and ma ny parts of Africa. In South Africa, P. falciparum is the most common form of malaria, with it causi ng 99% of the infections, and the remaining 1% being caused by P. malariae Found primarily in subtropi cal and temperate areas, P. malariae was the most common form of malaria in Europe, and the number of infections from this species fluctuated significantly throughout history, although they are rare today (Markell, Voge, et al. 1992:102). Finally, P. ovale is limited to Africa and certain parts of southeastern Asia, with some reports of this species in South America and other parts of Asia (Markell et al. 1992:101). There are about 40 different types of mosquitoes in the Anopheles genus that transmit malaria, with only about 30 of them causing the most concern (Breman 2001:4;
13 Bruce-Chwatt 1985). Since the transmission of the disease depends upon factors including the host, the vector, and the environment, all of which vary in numerous ways by the species of the parasite, the protozoa have adapted to many environments. Drug resistance has occurred in some instances (Sachs and Malaney 2002:680). 2.3 Disease Process and Symptoms The process of a malaria infection is co mplex (Figure 2.1), consisting of several growth phases (the following description has been compiled from Aufderheide and Rodrguez-Martn 2005:229-230; Carter and Mendis 2002:566-569; Markell et al. 1992:96-100; Sinden and Gilles 2002). Explaining it from the aspect of human-to-human transmission, the process begins with a mo squito biting an infected human. The first phase of growth is called sporogony. The blood cells in the host that are of interest are the ones containing the male and female sexual cells (gametocytes) of the Plasmodium genus, which are ingested by the mosquito. In the stomach of the mosquito, the acids cause hemolysis (the rupturing of the red blood cells causing their contents to be released in the surrounding fluid) of the humans blood cells, releasing microgametocytes (male sex cells) that fertili ze the macrogametocyte (female se x cells). After fertilization, a cystic structure, called an okinete, cont aining a mass of sporozoites forms. This structure becomes more round (ocyst) and attaches to the wall of the mosquitos stomach. The cyst eventually ruptures rel easing sporozoites that migrate into the mosquitos saliva and are injected into the blood of the next human the mosquito bites. About 40 minutes after a human is bit by an in fected mosquito, the sporozoites migrate to the liver, or hepatic, cells where they multiply asexually. During this hepatic phase, the
14 Figure 2.1. The malaria life cycle and infection process (courtesy of CDC/Alexander J. da Silva, Ph.D./Melanie Moser from the Centers for Disease Control Public Health Image Library) human does not have many, if any, symptoms Instead, the sporozoites are multiplying, and these eventually become merozoites (cells produced during asexual reproduction of the protozoa). Depending upon the species of Plasmodium that has infected the human, the incubation time varies from two ( vivax ) to six weeks (malariae ). After the incubation period is complete, the merozo ites are released from engorged liver cells, and they enter the bloodstream. There is also variation in the disease process af ter this, depending upon the species with which the person is infected. For example, the P. ovale and vivax types will reenter the liver cells and repeat the hepatic stage, creating a resting stage of the parasite, known as a hypnozoite. After seve ral weeks or months, the hypnozoites can become reactivated, creating another infection. Therefore, it is possible for a relapse to
15 occur after treatment. This does not occur with P. malariae and falciparum which only infect the human in a single cycle; subsequent infections are caused by the bite of another infected mosquito. After the cells enter the bl oodstream, they attach to receptor sites on the blood cells, specifically erythr ocytes, or red blood cells. For example, it is known P. vivax uses the Duffy Fy6 RBC antigen as a receptor site; however, the receptor sites for the others are not known, but are probably unique for each species. Once in the red blood cell, the merozoite matures into a ring form, or tr ophozoite, which is the feeding and growing stage. There it divides, a process known as schizogony, and cytoplasm accumulates around each of the nuclear fragments, which creates new merozoites. This process uses 25 to 75% of the red blood cells hemoglobin (Wyler 1983a, b), and products resulting from this process are toxic to the red blood cells (Orjih et al. 1981). The red blood cells then rupture, releasing more merozoites that infect other red blood cells, repeating this intraerythrocytic cycle. Gametocytes also appear in the red blood cells and circulate throughout the bloodstream, infecting a mosquito when it bites. The length of the intraerythrocytic cycle is constant for each species; it is 24-48 hours for P. vivax, 48 hours for P. falciparum and P. ovale, and 72 hours for P. malariae It then takes about a week for the cycles to become synchronized, resulting in a massive and simultaneous destruction of red blood cel ls and merozoite release. When this happens, the symptoms of the clinical descrip tion of malaria occur. These signs include a sudden high fever, headache, muscle pains, and chills, all of which subside as the merozoites reenter the erythrocytes and re occur a few days later, depending upon the length of the species intraerythrocytic cy cle. Eventually, after several cycles,
16 gametocytes are formed rather than merozoite s. Mosquitoes ingest the gametocytes, and the cycle continues when the mo squito infects another person. 2.4 Acute and Chronic Malaria The severity of the clinical features of malaria vary by the degree of the infection (how many cells are being parasi tized) as well as other featur es of the type of species infecting the person, such as the age of the cells that can be infected and proteins created during the infection process. For example, P. ovale and P. vivax typically produce a milder form of malaria, while P. falciparum is usually fatal and cons idered to be the most dangerous form. P. falciparum has developed a resistance to drugs over the last 20 years (Breman 2001:3; Gupta et al. 1994; Trape et al. 1998), and in fact, our present knowledge of this species causes researchers to dia gnose malaria fatalities as resulting from P. falciparum although sometimes P. vivax is more fatal than assumed (Hume et al. 2003b:181). Diagnosis of malaria can be made from the symptoms, especially if a person has been in areas where malaria is present. The type of infection can be confirmed by identifying parasites in the blood (Breman 2001:1). At minimum, in acute malarial infec tions, a person infected with malaria will have no symptoms, which is known as asymptomatic parasitemia (Breman 2001:1; White and Breman 2001). Some of the resulting clinic al conditions from the infections include encephalopathy, cardiac arrhythmias, pulm onary insufficiency, renal failure (in P. malariae ), regularly spaced paroxysms, and spleno megaly (Hendrickse et al. 1972). If an enlarged spleen becomes injured and ruptures it can be a cause of death even in the milder cases of malaria. Cases of severe malaria may result in anemia, neurological
17 syndromes, or death (Breman 2001:1; White and Breman 2001). However, complications from malaria are usually limited to the P. falciparum species. These include cerebral malaria (resulting in a severe headache, fa tigue, confusion, and eventually coma), anemia, renal disease, Blackwater fever (a s yndrome which leads to severe anemia and may also be associated with vivax and quartan malarias), dysentery, algid malaria (hypotension and impaired vascular pe rfusion), pulmonary edema, Tropical Splenomegaly Syndrome (TSS unusually l ong-term swelling of the spleen), hyperparasitemia, and hypoglycemia (Markell et al. 1992:105-108). Children with malaria commonly experience severe anemia hypoglycemia (both of which are also common in pregnant women infected with malaria), respiratory distress, and coma. Malaria may also cause learning and behavioral disorders (Anderson 1927; Breman 2001:1). Malaria does not affect the gross appearance of skeletal remains directly; however, related conditions, such as acquire d and inherited anemias, can result in changes to the skeleton. These changes include porosity in the outer table and orbital roofs of the cranium, osteoporosis, and other morphological conditions, all of which are discussed in detail in Chapter 3. In cases where the individual is not treat ed, the fevers associated with malaria generally become less intense and irregular. W eeks after the initial infection, people enter the chronic stage of the disease, where they maintain a low level of infection. Although this stage is marked by moderate anemia and fatigue, one can generally return to performing normal activities. An individual with a chronic infection may have acute recurrences or may become infected with another strain of malaria if living in or visiting an area where malaria is present (Aufde rheide and Rodrguez-Martn 2005:231).
18 2.5 Malaria and Other Diseases Besides complications associated directly with malaria symptoms, there are also indirect problems from this disease (Sallares 2002:122-139). For example, lowbirthweight babies have been born to mothers with malaria, which causes the infants to have a lowered immune response to other infections, thus making the infants more susceptible to conditions such as gastro-int estinal diseases that may result in death (Desowitz 1992:118; Urban et al. 1999). It shou ld be noted that malaria itself can result in dangerous gastro-intestinal respon ses (De Korte 1899; Murty et al. 2000). Individuals infected with malaria may also be infected with other diseases. The interactions between these diseases can ha ve various affects. Sallares (2002:123-139) outlines effects upon the pathogens or the host as being synergistic, antagonistic, or neutral, and he notes that human adaptations to one disease may also cause a decreased resistance to another pathogen. An example of an antagonistic relationship is that between malaria and syphilis; syphilis cannot tolerate the heat produced by malarial fevers (Fraser 1998). The comorbidity of malaria and smallpox was observed by Lapi (1749) to have resulted in a neutral relati onship, with both diseases running their courses separately. The interaction of malaria with t uberculosis is more complex. First, repeated malarial infections (even if asymptomatic) can compromise the immune system of the host making the individual more susceptible to tuberculosis (Enwere et al. 1999). Interestingly, when individua ls suffering from chronic malaria become subsequently infected with pulmonary tuberculosis, the result is a slow course of t uberculosis that often includes the sclerosis of tissu e; on the other hand, individu als with tuberculosis who
19 subsequently develop malaria have aggravated tuberculosis symptoms that result in a rapid development of tube rculosis (Collari 1932:324). 2.6 Biological Adaptations Since malarial species use up to 75% of the hemoglobin in a red blood cell before hemolysis occurs and frees the newly-formed parasites (Wyler 1983b:935), a condition impairing the production or avai lability of hemoglobin to the parasite will have an impact upon the ability of the individua l to contract malaria. There are many adaptations to malaria that accomplish this (see Livingst one 1967, 1971; May et al. 2007). One example of such an adaptation is hemoglobin S, in which the chemical composition of the hemoglobin makes it more difficult for merozo ite growth to occur in the instance of P. falciparum infections (Allison 1954). Sickle cell disease in its homozygous state (denoted as HbSS), results in misshapen red blood cells (resembling a sickle blade). This state creates a dangerous condition because the mi sshapen cells are destroyed prematurely causing the clogging of capillaries and resulting in impaired blood flow to organs (Aufderheide and Rodrguez-Martn 2005:232; Gilles et al. 1967). HbSS has historically resulted in death during childhood if left untreated. Heterozygous HbSA (HbA = normal hemoglobin) results in both types of hemogl obin being produced, creating a resistance to malaria, mainly P. falciparum (Markell et al. 1992:113), with out contracting sickle cell disease. Other adaptive conditions are less well known. These include alterations that produce a similar effect, and are HbC (Modia no et al. 2001), HbE, HbF, thalassemias, and glucose-6-phosphate dehydrogenase (G6P D) (Allison and Clyde 1961; Flint et al. 1986; Ruwende and Hill 1998; Ruwende et al. 1995; Tishkoff et al. 2001). Another
20 example is the lack of the Duffy Fy6 RBC antigen. This is hypothesized to be an adaptation to vivax malaria. In western Africa, this species of mala ria does not commonly occur, probably because of the almost univers al lack of the Duffy antigens (Miller et al. 1976) one of which acts as a receptor for the disease. Allison and Clyde (1961) note the lack of Duffy Fy6 RBC antigens also offers protection from P. falciparum Aufderheide and Rodrguez-Martn (2005:232) state the recognition of sickle cell disease as a balanced polymorphism is well documented but has not yet won universal acceptance. They also think th ese other aforementione d genetic conditions have been researched even less, and their processes as balanced polymorphisms are still questionable. In particular, they note that although thalassemia and malaria are related geographically, it is not unders tood how thalassemia impacts a malarial infection. On the other hand, Allen et al. (1997) assert thalassemia has shown to provide protection from other infections as well as malaria. Thalassemia, also referred to as Cool eys anemia, Mediterranean hematologic disorder, and Mediterranean hemopathic syndr ome (Chini and Valeri 1949), is a globin deficiency, which results in the hyper-produc tion of red blood cells (see Thein 1993 for a detailed explanation of the biochemical processes related to the inheritance of thalassemia). There are different types of th alassemias, and they are named by the globin chain affected (alpha or beta beta being the most common), with major referring to the homozygous form of the condition, intermediate referring to a homozygous form that is less deleterious, resulting in symptoms but requiring no medical intervention, and minor the heterozygous, and less serious, state (Auf derheide and Rodr guez-Martn 2005:347). This inherited disease is found in approximately sixty countries. There is a high
21 prevalence in the Mediterranean, and it continue s into the northern and western regions of Africa, and from the Middle East through India and into Southeast Asia (Amini et al. 2007). About 150 million people worldwide have be ta-thalassemia genes. The prevalence in Sardinia and Sicily for beta-thalassemia ranges from 10 34% (Amini et al. 2007:36; Thein 1993:169). The area where thalassemia is most common is known as the thalassemia belt (Figure 2.2) (Amini et al. 2007:36; Thein 1993:169). Elevated frequencies of these balanced polymorphisms are found only in populatio ns with at least several hundred years of exposure to malaria; thalassemias and G6PD deficiency are usually the first genetic responses to emerge (Carter and Mendis 2002). While all these polymorphisms discusse d here might offe r protection from malaria, they are also producing other diseases such as thalassemias, sickle-cell disease, G6PD deficiencies, and ovalocytoses, causing further health impacts (Carter and Mendis 2002:574). These impacts are projected to incr ease in the future as childhood mortality Figure 2.2. Map identifying the lo cation of the thalassemia belt. Sardinia is identified with an arrow (www.outline-world-map.com).
22 decreases with improved medical conditions ; however, although me asures are being taken to provide information about these diseases they still are not a health care priority. In addition, information and educational progr ams concerning these diseases, which take considerable time to establish, will have to be provided to poorer countries where these diseases are most prevalent (Weatherall and Clegg 2001). 2.7 Cultural Adaptations Breman (2001:2) describes the effect s of malaria upon the individual and the community as having intrinsic and extrinsic de terminants. The intrinsic determinants that are the most important include host (hum an) immunity, paras ite species, anopheline longevity, and avidity for humans and the extr insic factors that are the most important are climate (mainly rainfall), economic conditions (poverty), political commitment, and effectiveness of control and pr evention efforts. Culture is a player in the disease process in two aspects. First, societies interact w ith their environment to change the risk of particular diseases (often times these acti ons are not deliberate), and then societies construct an understanding of diseases while treating them with medicine (Inhorn and Brown 1990:110). For instance, ear ly historic documents reco rd the relationship between illness and environments that have been modified by humans. This relationship still exists today. Deforestation for agricu ltural and development purposes often results in marshy areas conducive for the life of the mosquito vector (Hume et al. 2003b:182; Livingstone 1958; Mitchell 2003:174; OSullivan et al. 2008). Frequently, these deforested areas are close enough to locations where people are living in a high enough c oncentration to allow the disease to be transmitted. Construction also plays a role in the infective process of
23 malaria. An example is the construction of dams which results in groups of humans being moved to areas that sustain malaria parasite s. Hughes and Hunter (1970:479-480) refer to this as developo-genic malaria. While the rapid diagnosis of malaria (w hen facilities are available) and the improvement of hygiene and public health (e .g., filling of swamps, open ditches, and the removal of standing water) are essential for reducing the risk of malaria infections (Breman 2001:6), cultural adaptations, esp ecially belief systems, have roles in understanding, sustaining, treating, and eradicating malaria. It is suggested (Armelagos and Dewey 1970; Dunn 1976; Heinrich 1985; MacCormack 1984) that cultural adaptations be integrated with antimalari a campaigns and public health initiatives, preferably in an interdisciplinary manner. Some observed adaptations that counteract the disease include making modifications to build ings. For instance, homes constructed on stilts above the fly zone of mosquitoes, which carry the malaria parasite, reduce the number of infections (May 1958). In addition, Ewald (1994:42-43) notes that the construction of brick and mortar housing w ith appropriate roofi ng, screening, and air conditioning (as opposed to thatch housing) has greatly reduced the transmission of malaria. Another illustration of an adaptation to malaria is inverse transhumance (utilizing lowlands for herd grazing during th e winter, while maintaining residence in the uplands) when used in conjunction with a nuc leated settlement pattern (Brown 1981a). Some other examples of cultural adaptations to malaria include the use of numerous plants as antimalarial drugs (see Et kin 1997); in fact, it has been suggested that domesticated plants and mala rial parasites coevolved (Jac kson 1997:179). Besides plants that are used for drugs, fava beans ( Vicia faba) is an example of a food that increases the
24 effectiveness of the G6PD deficient genotypes; however, consumption of these beans can also result in favism (see Turrini et al. 1997). Favism is a life-thr eatening condition that causes hemolysis in certain circumstances. It mainly affects children (Greene 1997:209211). Greene (1997:227) also discusses dietar y patterns that reduce the number of antioxidants for the individuals to benefit from the antiparasitic oxidant effect of fava beans. Western responses to malaria included eradication efforts through the use of dichloro-diphenyl-trichloroethane (DDT) in the 1950s. These efforts were highly effective in places such as Europe (including Sardinia). However, the use of DDT is a concern from an environmental and health standpoint (Beard 2006), and since these efforts started, some mosquitoes have become DDT-resistant (Baleta 2009; Porter 1997:471-472). Today there are several anti-mal arial drugs, such as quinine. While these drugs are effective, they are also producing drug resistant forms of the disease. Some of these drugs also have other repercussions, such as hyperinsulinemia from quinine use (Breman 2001). It has been debated if world-wide attempts to eradicate malaria should be made, with the cost and effectiveness of these methods, as well as the emergence of insecticide resistance, population movements, and war weighing heavily into the decision not to attempt it (Porter 1997:471-472). Although research into vaccines and new drugs continue, the issue is now main ly one of control. For example, current control practices put in place by the World Health Organizati on include the distribution of mosquito netting, clothing, and bedding treated with inse cticides (Rowland et al. 1999), which have produced favorable results (Alonso et al. 1991 ; Guillet et al. 2001), However, the best preventative methods involve breaking the human-vector cycle (Markell et al. 1992:115-
25 122). This is accomplished through the management of the vector mosquitoes, which is established from the knowledge of all possibl e methods of control, including technical considerations for each of the control methods and an understanding of the human populations, mosquitoes, and environmental fact ors of the area. This control philosophy also relies on the responsibility, particip ation, cooperation, and the community of action (Dunn 1983). 2.8 History and Debates Most of the research concerning the dissemination of this disease is interpreted from the descriptions made in historic documents. Hume et al. (2003b:181) note that documentations of malaria-like symptoms have been found dating to 1500-800 BC in India and China, which include the notation of the use of an anti-malarial agent to treat the disease. Evidence of malaria symptoms in the Egyptian Papyrus Ebers (ca. 1500 BC) made the association between mosquitoes and malaria symptoms, and hieroglyphs at Dendera (Upper Egypt) associate fevers with the flooding of the Nile (Jackson 2000:280). This Egyptian documentation has be en corroborated by the analysis of mummified remains (Hume et al. 2003b:181; Nerl ich et al. 2008). References to malaria by Greeks and Romans were later, dating to about 500 BC, according to Aufderheide and Rodrguez-Martn (2005:234). Russel (1952:93-96) provides a good summary on the etymology and etiology of malaria in western history. The word malari a comes from the medieval Italian words mala and aria ( malaria ) meaning bad air. Other terms us ed to describe this disease included malaqua (bad water) and paludism (stemming from palustris the Latin word for
26 swamp). As early as the first century BC, people considered the etiology of malaria. Varro linked marshy areas as being conducive to the growth of tiny organisms (that could not be seen by the human eye), which ente red the nose or mouth and caused fevers (OSullivan et al. 2008). Subsequently, in the first century AD, Columella expressed concern about the presence of buildings and highways near marshes because he thought steam and insects from these areas caused dise ase in humans. In fact, malaria in Italy has been reflected upon so much that Russel (1952 :99) writes, so famous have been the Italian activities in malariology that for many years and up to the present time there has been a stream of students coming from overseas to learn from Italy One can hardly find an outstanding malariologist in the worl d today who has not visited Italy for the purpose of observation or formal tr aining in this special field. Historically, before the di agnosis of malaria was made, the disease was described by the length of the febrile cycle. For ex ample, in 350 BC, Plato taught that quotidian fever (24-hour cycle/vivax) was related to air, tertian (48-hour cycle/vivax for benign malaria, falciparum if malignant or aestivoa utumnal or subtertian) to water, and quartan (72-hour cycle/malariae malaria) fever to earth. Ovale malaria ha s been only recently described, and thus does not ha ve a descriptor from a fe brile cycle (Markell et al. 1992:96). Because the symptoms of malaria (f ever with periodic reoccurrences and splenomegaly) are similar to those of other diseases, and the symptoms vary (such as the lack of recurrent fevers with some speci es, or the variation in the times between recurrence), the disease was difficult to diagnose before medicine was able to link symptoms with specific etiological agents (Aufderheide and Rodrguez-Martn 2005:333).
27 It is not certain who made the first mosquito-malaria connection, as people have believed mosquitoes caused fevers in humans for centuries in areas such as Africa and Asia. However, in the late 1800s the conn ection between mosquitoes and malaria was solidified. Ross was the first to hypothesize that mosquitoes are a disease vector for the malaria parasite. Because his work was focu sed on the wrong species of mosquitoes, it took five years for him to discover parasites in the correct species. At the same time, in Italy, Grassi, Bastianelli, and Bignami had non-malarious volunteers participate in controlled experiments, in which they either allowed themselves to be bit by infected mosquitoes or were exposed to marsh airs in highly malarious environments. Ultimately, because of these researchers, the connec tion between mosquitoes and malaria was accepted (Russel 1952:96-97). Today, researchers are trying to establish a chronology of the interaction between malarial parasites and humans. Many debates exist regarding the evolution of the Plasmodium sp., in particular P. falciparum and its transmission between human populations throughout history (Hume et al. 2003b:182). There are four questions regarding the origin of P. falciparum The first is when the parasite diverged from P. reichenowi There are two thoughts on this. One is that it diverged at the same time as did the host lineages, about ten million years ago (Escalante et al. 1995; Hey 1999). The other is that because it is so virulent, that it must have diverged more recently, perhaps within a few thousand years, because it woul d have otherwise adapted to humans as a host, as dead hosts are not useful to parasites (Ewald 1994; Sallares and Gomzi 2001:197). Second, the presence of human adapta tions to this species suggests it may have been endemic in hunter-gatherer popul ations (Cavalli-Sforza et al. 1994), while
28 others feel, after reviewing historic document s, that it began affecting humans only about 2000 years ago (Bruce-Chwatt and de Zulueta 1980) Third, it is not clea r if the earliest civilizations experienced ende mic malaria. Sallares and Go mzi (2001:197) argue the first convincing documentation of malaria was made in the fifth century BC in the Hippocratic corpus. The final question invol ves the population structure of P. falciparum ; that is, is there enough genetic va riation to exclude th e possibility of the organism passing through a bottleneck within the last 20,000 years? Although the fossil evidence is not complete, the distribution of current strains of malarial organisms, and the high level of ad aptation needed to produce the intricacy of the relationship between the hosts and insect vectors, indicate malaria is one of the most ancient human parasite s, with the genus Plasmodium emerging hundreds of millions of years ago (Escalante and Ayala 1995). Resear ch by Rich et al. (2009) supports the hypothesis of P. falciparum evolving from the species P. reichenowi which infects chimpanzees; however, it not known when or where the four most common types of malaria began infecting humans. Because of the similarity of the strains infecting humans to those infecting apes (Rich et al. 2009), it has been speculated the origins of malaria are in Africa. Cockburn (1963:2) suggests malaria coevolved in Africa w ith humans. Rich et al. (1998), through the analysis of DNA, suggest the P. falciparum species of malaria began between 24,500 57,500 years ago, or more likely before then, coinciding with environmental changes, increase of the mosqu ito vectors, and the eventual advent and spread of agriculture. They also think non-African cases of P. falciparum malaria occurred only within the la st 6,000 years. This interpretation is also supported by Volkman et al. (2001). Hughes and Verra (1998) disagree and note there is evidence
29 supporting otherwise, formulated from a mo re comprehensive genetic analysis (using more loci). More recent gene tic studies demonstrate that P. falciparum has even older origins. Studies by Mu et al. (2002) indicat e the most common recent ancestor occurred around 100,000 180,000 years ago, and research by Hughes and Verra (2002) indicates the most common ancestor is even older 150,000-200,000 years ago. Still others (e.g., Hartl et al. 2002) continue to take a more conservative stance, and believe that with advances in genetic research more effective studies will be conducted so we will be able to determine how long ago P. falciparum and other malarial species originated. Some genetic evidence indicates there were multip le waves of expansion, both ancient and more recently with the Neolithic Revoluti on, spreading separately from Africa to Southeast Asia and South America (Conway et al. 2000; Joy et al. 2003) resulting in a population bottleneck of this species (Hume et al. 2003a). Coluzzi (1999) notes that because of the high mortality rate linked with P. falciparum it is not likely the parasite is that old, otherwise it would have become extin ct, and it is likely the strain present today is modern, and probably did not occur in It aly until the last two centuries BC. The virulence of P. falciparum has been argued to be the result of the parasite having insufficient time to adapt to its human host (assuming parasites evolve toward commensalisms); however, to estimate the para site has only been infecting humans for 10,000 years is a flawed argument because 10,000 years is a long time for a protozoan parasite to evolve (Ewald 1994:43). Correlati ng genetic information about malaria, as well as human adaptive responses and envi ronmental conditions is necessary for understanding the history of ma laria and developing effective methods for combating the disease (Hartl et al. 2002 ; Hume et al. 2003b:188).
30 De Zulueta (1973, 1994) notes that temperat ures equal to about those of today began about 10,000 years ago at the close of the Pleistocene, and it is likely that this is when malaria (with the exception of P. falciparum because of the insect vector not responding to the African malarial species) en tered Europe (Coluzzi 1999; Livingstone 1958). Malaria may have spread earlier in th e Levant and parts of Asia because the temperature did not change as much in th ese areas. Experiments conducted by Ramsdale and Coluzzi (1975) indicate it is unlikely P. falciparum infections in Europe were caused by infected humans from Africa, as the mos quitoes they have experimented with do not carry the disease, but rather it is likely Eur opean malaria cases are the result of a mutation of P. falciparum A review of historical documents and paleoclimatological data (Bruce-Chwatt and de Zulueta 1980) indicates malaria probably was not present in Europe before 500 BC, fourth century BC in Greece (Jones 1967), beca use population concentrations were too low and the temperature was too cold to support the species of mosquito required to transmit the disease (Aufderh eide and Rodrguez-Martn 2005:234). Furthermore, these authors suggest that only the P. vivax and P. malariae forms were present, and only in southern Europe. However, Angel (1966, 1967) suggested in his res earch that malaria was indeed present during the Neolithic. He came to this conclusion by examining skeletal remains from the eastern Mediterra nean, which he notes coincides with areas where P. falciparum and thalassemias were pres ent (although at the time of his publication, there were no autopsies or radiog raphs of heterozygotes with thalassemia or sickelemia).
31 Angel (1966:760) thought th e skeletons of young childre n with severe porotic hyperostosis (lesions found on th e outer table of cranial bones resulting in the thickening of the bone from the expansion of the diploic layer resulting in responses ranging from a few, small porous lesions to exposure of trab eculae; the etiology of porotic hyperostosis is discussed in Chapter 3) were those whic h were homozygous for thalassemia, as they would likely die early in life, while the adults with porotic hyperostosis were heterozygous and the degree of porotic hyperostosis reflected the degree of the anemia resulting from a falciparum malaria inf ection. Those homozygous against thalassemia would not have been likely to survive a falciparum infection long enough to experience bony responses. To support his arguments with prehistoric datasets, he used demographic, archaeological, and environmental data. He compared these bony responses to changing sea levels during the Neolithic under the premise that the changing sea levels would provide breeding areas for the vectors. His hypotheses were checked by studying historic Greek populations with documentation of malaria. Here, he noticed the rates of porotic hyperostosis and decreased stature fluctuated w ith the incidence of malaria. The examination of the climate by Bruce-Chwatt and de Zulueta (1980) does not agree with this entirely. Furthermore, the pr otection afforded by the heterozygous state of thalassemia is supported by the presence of falciparal malaria; they believe these high correlations are found only in Italy and Sard inia (Bruce-Chwatt and de Zulueta 1980). A more recent interpretation by paleopathologist s Capasso and Di Tota (1995) support the hypothesis that P. falciparum was established during th e deforestation accompanying agriculture in the Old World during the Neolithic.
32 2.9 Chapter Summary Although the timing and nature of the exact introduction of malaria to Europe is still debated, it is possi ble that prehistoric populations in Sardinia were exposed to one or more species of malaria. Verifying if this disease was present during prehistory can result in a better understanding of th e timeline of the spread of this disease, the nature of the relationship between the pathoge n and host, the lives of preh istoric (in this case Middle Bronze Age) Sardinians, and human behavior. From the information presented in this chapter, evidence of malaria observable in a population can be used to create conditional statements to test the hypothesis that malaria was pr esent in a population. These conditional statements include: 1. if a population shows evidence of thalasse mia, then it is likely they were subjected to malaria (for se veral hundred years, possibly); 2. if tuberculosis is present in an area with endemic malaria, then it is probable some individuals will have had a prolonged disease process resulting in the sc lerosis of tissue; 3. if a population is subjected to malari a, then there may be evidence of cultural adaptations, such as structures limiting contact with mosquitoes or the use of medicinal plants or foods (e.g., fava beans); 4. if an individual was infected with malaria, there may be gross skeletal evidence of related conditions, such as acquired and inherited anemias. Since this research is bioarchaeological in nature and includes th e analysis of human remains from Sardinia, knowledge about human bone, previous pale opathological studies concerning malaria, and the prehistory of Sardinia is required to develop a
33 comprehensive methodology to address the quest ion: was there malaria in prehistoric Sardinia?
34 Chapter 3: Paleopathology, Archaeology, and Malaria The use of skeletal collections to study a disease in prehisto ry provides us with two benefits. One, it establishes the antiquity and evolution of humans and the disease. The antiquity of a disease can be establishe d through its correlation with the results of archaeological dating techniques, and the evolution can be studied through the analysis pathogenic DNA. Two, it places the disease in a context that includes the physical and cultural aspects of humans. This allows us to understand the relationship of the disease and biocultural adaptatio ns more completely (Inhorn and Brown 1990:93). The coevolution of host, vector, and pathogen can be better understood by examining changes in the structure of their DNA over time. The knowledge of chemical and cellular components of bone, as well as factors such as growth and bony responses to disease, forms the basis of paleopathological analys is and theory. When this information is examined within the framework of a specifi c disease, hypotheses can be developed and tested. 3.1 Bone Many texts provide good desc riptions of the compositi on and attributes of bone (e.g., Marks Jr. and Odgren 2002; Ortner 2003; Steele and Bramblett 1988; White 2000). When assessing disease processes, paleopa thologists often focus on bone remodeling, which is the result of the de struction and growth of bone ce lls. Since disease in skeletal
35 tissue is comprised basically of these two processes, this lim its the responses that osseous tissue can demonstrate; that is, differe nt processes can produce similar results. On the chemical level, bones are compos ed of two types of materials, collagen and hydroxyapatite. Collagen is a protein, a nd it makes up 90% of th e organic portion of bone. In bone, collagen forms flexible fibers that are hardened by hydroxyapatite. Hydroxyapatite, a form of calcium phosphate is what makes bones hard and rigid. From these, there are two structural type s of bone tissue, cortical (compact) bone and trabecular (spongy or cancellous) bone (Figure 3.1). Cortical bone is the hard, dense portion covering the shafts a nd external surfaces of bone s. Trabecular bone is the lightweight, honeycomb-like structure under cort ical bone, and it occurs in long and short bones, flat bones, vertebral bodies, and other bony protubera nces (e.g., tendon attachment sites). Although both types of bone material are identical on the cellular and molecular level, they differ in function. Cortical bone provides protection and the strength for mechanical functions. Trabecular bone, the site of hemopoietic (blood-forming) tissue, is Figure 3.1. Histological schematic of cort ical and trabecular bone (U. S. National Institutes of Health 2010)
36 responsible for metabolic functions, such as the production of red blood cells, white blood cells, and platelets. The nourishment of trabecular and cortical bone occurs in two different ways. Trabecular bone is nourished through blood vesse ls on the surface. Cortical bone does not receive nourishment this way, and it compensate s for this by the formation of canals and canaliculi (small fluid filled canals) in what is known as Haversian systems. A cross section of cortical bone looks similar to tree rings. These rings are known as Haversian lamellae, with each one of the lamellae containing bundles of collagen fibers. These collagen fibers are arranged in different directions in adjacent lamellae adding strength to the bone. Within these cross sections are bundles which re semble trunks, which are the Haversian systems or secondary osteons Haversian systems are about 0.3 mm in diameter and are three to five mm long; they run parallel to the bone of which they are a part and form the basic structure of compact bone. In the center of these systems is a core, called the Haversian canal, which c ontains blood, lymph, and nerves. Off the Haversian canals are Volkmanns canals, wh ich are smaller and jut off at right and oblique angles. Volkmanns canals go th rough the bone tissue a nd link the Haversian canals together from the periosteal to endos teal surfaces. This network supplies the cells with blood and lymph. In the lamellae, there are small cavities, lacunae, which contain osteocytes. These cells are nour ished via canaliculi that link Haversian canals to lacunae in adjacent lamellae, or they link one lacuna to another. Osteoprogenitor cells are responsible for bone growth and maintenance and include osteoblasts, osteoclasts, and bone-lini ng cells. These cells perform five functions: produce protein in the bone, stimulate mineralization, maintain bone tissue, resorb bone,
37 and contribute to mineral physiology. Os teoblasts, which are responsible for the production of the bone matrix and its regul ation, are usually located beneath the periosteum, and they deposit pre-bone tissu e, a material called osteoid. Osteoid is collagen-rich and becomes ossified when hydr oxyapatite is deposited within the osteoid matrix. When osteoblasts are surrounded by this inorganic matrix, they mature and become osteocytes, which maintain bone tissu e. Bone tissue is removed by the third type of cell, osteoclasts, in a process called resorption. Bone-lining cells, which cover the surface of bones that are not being remodeled, ar e flat, inactive, and elongated; little is known about their function other than that they must be removed for remodeling to occur. Living bones are covered with a tissue called the periosteum, and the inner surface of the bone is lined with an amorphous membrane ca lled the endosteum. Both of these tissues have bone-forming cells that generate grow th as needed, for example, when the periosteum is traumatized. These membra nes are more active in younger individuals because bone cell formation slows as one ages. During the embryonic stage, osteogenesis and ossification occur in two ways. The first is intramembranous ossification, whic h is the formation of bones by apposition on tissue within a connective tissue membrane in the embryo. Most bones, however, form by endochondral ossification, which is bone form ation from cartilage precursors (Figure 3.2). This is referred to as the cartilage mode l. Cartilage is composed of three different types of cells, chrondrocytes, chrondroclasts and chrondroblasts, which are analogous in function to the osteocytes, osteoclasts, and os teoblasts in bone. There are three different types of cartilage: hyaline cartilage (ossifies into bone), elastic cartilage (e.g., flexible portion of ears), and fibrocartilage (stabilizes joints). Both the intramembranous and the
38 Figure 3.2. An interpretation of bone growth from a cartilage model (U.S. National Institutes of Health 2010) cartilage model processes result in the same bone tissue being formed. When the cartilage model is penetrated by blood vessels, starting at the nutrient foramen, ossification occurs. On long bone cartilage models, a membrane (perichondrium) surrounds it. This becomes the periosteum when osteoblasts begin to de posit bone along the cartilage shaft. During the prenatal period, the first type of bone tissue that deve lops is woven bone, also known as immature bone or coarsely bundled bone. Woven bone is considered a primitive form of bone; however, it also occurs later in life. Because it has proportionately more osteocytes (living bone cells) than mature, or lamellar, bone, it is found during fracture repair and in bone tumors. Microscopically, woven bone is coarse and fibrous, and it has collagen fibers distributed randomly in bundles. This bone, typically ephemeral in nature, is replaced with mature, or lamellar, bone tissue. In contrast to woven bone, lamellar bone is orderly in nature, and it is genera ted more slowly. Lamellar bone makes up both the cortical and trabecular portions of bones.
39 Osteoclasts on the endosteal surface re move bone, and in the periosteum, osteoblasts deposit bone. Eventually, as the individual grows the diameter of the bone changes because of the deposition by oste oblasts. This pattern of bone removal and deposition allows the shaft diameters to en large during growth, while maintaining the proportion of the medullary cavity (the space in the diaphysis where marrow is stored). The bone grows in length when the cartilage between the metaphyses (rough ends of the long bone) and the epiphyses (or the secondary centers of ossification), also known as the growth plate/epiphyseal plate, are replaced by bone. This pr ocess pushes the epiphyseal plate farther from where the bone began to ossify thus causing the bone to lengthen. When the cells of the epiphyseal plates stop dividing, the primary and secondary ossification centers become fu sed. Further remodeling occurs during this process, which results in flaring at the ends of the long bones. The human skeleton begins with approximately 800 ossification centers at bi rth and ends with 206 bones in the average adult skeleton. The skeletal system continues to change dramatically as the individual matures throughout adulthood. This process, where old bone tissue is removed by osteoclasts and replaced by osteoblasts, is called remodeling (or sometimes modeling). Remodeling allows for the maintenance or change of bone size and shape during growth periods and repair of injuries. In remodeling, when bone resorption is equal to the amount of bone formation, it is called coupling; in other stages, such as earl y growth or during elder years when these are not equal, it is referred to as uncoupling. In life further modification occurs when the position of the bone cells change to adapt to mechan ical stress; this is known as Wolffs law.
40 Diseases can also affect the morphology of bone at the macroscopic and microscopic level. Ortner ( 1992:5-7) cautions that acute di seases result rarely in bony responses, and what is observed in the skeleton is usually the result of a chronic disease, caused generally by bacteria. Different diseas es may result in similar bony lesions, as bone can only respond in a limited number of ways. 3.2 Analyzing Skeletal Remains in Po pulations Affected by Malaria Although malaria itself does not result in any direct bony response in the skeletal system, it is related to two conditions that do. The first condition is anemia (StuartMacadam 1992a:164). Current medical and epid emiological research can provide some insight about malaria and anemia. Some (Gandapur et al. 1997) report that P. falciparum does not have a characteristic clinical or haematological condition, yet they acknowledge it is a major cause of severe anemia in the malaria endemic area of Kenya. Others (Wickramasinghe and Abdalla 2000) note th at there are changes (at least on the microscopic level) in the bl ood and bone marrow in cases of malaria. A study with 234 subjects conducted by Nyakeriga et al. ( 2004:439) examines the complex relationship between iron-deficiency anemia and malaria in sub-Saharan Africa and concludes that iron deficiency was associated with a protection from mild clinical malaria. In fact, iron supplementation has also been shown to be dele terious in cases of ma laria, leading these researchers to conclude that iron supplementation should be contraindicated in malarious regions. Research by Gandapur et al. (1997) also shows in some instances prolonged infections of P. vivax and P. falciparum malaria result in hype rcellular bone marrow pathology associated with iron-deficiency an emia. Bone marrow has also been described
41 as hypercellular in patients with malarial infections by other res earchers (e.g., Metha et al. 1996). The second condition is related to inherited hemolytic anemias. For example, thalassemia can be observed in the skelet on because it produces a specific pattern of skeletal responses. This conditi on is also very common in Sard inia, so it is important to be aware of thalassemic indicators when an alyzing prehistoric remains, as they can provide indirect clues about the possible presence of malaria. Ancient DNA analysis is another method used to examine the past. Besides providing descriptive information about human ge netic material, the analysis can be used to test hypotheses that are resolvable with aDNA from other organisms. For example, while conditions such as thalassemia may be detected using these methods, there is a chance Sardinian archaeologists and workers, who have handled the remains, may have thalassemia markers and th erefore their own DNA could contaminate the results. Therefore, aDNA analysis can be used to test for malarial DNA directly (Brown 2000:472). In addition, immunological techniques cr eated for diagnosing malaria in living individuals have produced some positive resu lts when applied to human remains from archaeological sites. Contaminants and false pos itives occurring in individuals with other immunological disorders have created questions about the reliability of this technique. Further research with these tests may result in more promising results. Malarial pigment, or hemozoin, is a bi omarker that has been used to identify malarial infections at the mi croscopic level in blood samples and gross-level autopsies. However, it has yet to be isolated from mummified remains or human bone tissue (only recently has it been isolated and detected from blood samples, see Aufderheide and
42 Rodrguez-Martn 2005, Nyunt et al. 2005, a nd Scholl et al. 2004; Sullivan 2002). The insoluble nature of this biocrystal and the unique signature it pr oduces when analyzed with laser desorption mass spectrometry (LDM S) provide a potential method to detect malaria in human remains from archaeological contexts. 3.2.1 Porotic Hyperostosis, Cribra Orbitalia, and Anemia Both porotic hyperostosis and cribra or bitalia were first described by Welcker (1888) as cribra orbitalia when he observed porosity in the orbit areas of frontal bones and on parietal and occipital bones. The term osteoporosis symmetrica was subsequently used by Hrdli ka (1914), Williams (1929), and Hooton (1930) to describe these conditions. Spongy hyperostosis (Hamperl and Weiss 1955) is another name for these bony responses. Porotic hyperostosis (Figure 3.3) is a condi tion that is found in cranial bones in which the outer table of the cranial vault thins or is completely destroyed (White 2000:528). The trabecular bone (inn er part), also known as the diplo, expands or appears in an area where it does not normally occur, and the outer layers of the cortical bone become porous. Bones that show this conditi on appear thick and s pongy with a coral-like appearance (Cohen 1989:107; White 2000:394). In some cases, a reaction to the periosteum results in a thickeni ng of the outer table of the s kull, without expansion of the diplo, and produces a similar condition (Ort ner 2003a:56). Microscopic examination can differentiate between expansion of the diplo or thickening of the out er table, and thus narrow the etiology of the condition. Cribra or bitalia (Figure 3.4) is the condition when it causes pitting in the orbital roofs (Cohen 1989:107; White 2000:394-524; Whittle and
43 Figure 3.3. An example of porotic hy perostosis on a parietal bone fragment from Serra e Sa Caudeba, cranium e2 (photo by author 2008) Figure 3.4. An example of cribra orbitalia on a frontal bone fragment from Serra e Sa Caudeba, cranium 3 (photo by author 2008)
44 Folkens 2005:320). These conditions occur sy mmetrically and are found primarily in younger individuals because it is the result of a childhood condition, typically anemia. The lesions are not as marked in adults as they are in children (Aufderheide and Rodrguez-Martn 2005:346-347; Holland and OBrien 1997:189). There is no evidence to suggest skull changes occur in adult indi viduals (in cases wher e these conditions are observed in adults, it is most likely it is a result of a ch ildhood episode after which the skull has not undergone complete remodeli ng) (Stuart-Macadam 1985:395). Adults, on the other hand, show marrow expansion in the medullary cavities of long bones and marrow producing masses (Aufderheide a nd Rodrguez-Martn 2005:350). Porotic hyperostosis and cribra orbita lia are the most common pat hologies found in prehistoric populations (Cohen 1989:107; St uart-Macadam 1989a, 1989b). While porotic hyperostosis and cribra orbi talia are familiar to paleopathologists, current pathology books rarely mention them (Henschen 1961:724). However, radiographs have produced images consistent with these conditions. A presentation of a hair-on-end appearance has been noted in th e X-rays of the crania of those living with congenital anemias, such as thalassemia, sickle-cell anemia, and spherocytosis. Polycythemia vera, congenital heart disease, hypoxia, and iron-deficiency anemia. Some cases of acute leukemia are also associat ed with the hair-on-end appearance in radiographs. Other examples of conditi ons producing these responses include: inflammatory processes, hemorrhagic processes (e.g., in scurvy, which presents in lesions on the greater wings of the sphenoid and adjacent bone, as well as th e orbital roofs), and anemia related to other causes, such as para sites (Ortner et al. 1999; Ortner and Erickson 1997; Schultz 2003:89). In rare instances, porotic hyperostosis and crib ra orbitalia have
45 been observed in cases of rickets and tumo rous processes (Schultz 2003:89). StuartMacadam (1989a:219) notes that scurvy, rick ets, and iron-deficiency anemia are interrelated; that is, iron-deficiency anemia is often associated in cases of rickets or scurvy, and in some cases, all three conditions occur simultaneously. However, historic documents suggest that scurvy does not fre quently occur before medieval times, and rickets is only common after the Industrial Revolution (Stuart-Macadam 1989a:219). To identify the cause of porotic hyperost osis and cribra or bitalia accurately, microscopic or radiographic examination is necessary (Schultz 2003:105). StuartMacadam (1987b) suggests comparing clinical Xrays with those taken on remains from archaeological contexts to aid in the id entification of the c ondition causing this bony response. Without corroborating informati on, Schultz (2003:105) recommends restricting their interpretations to non-specific indicators of stress, rather than attributing them to a specific disease. Rothschild (2000) notes today it is rare that porot ic hyperostosis results from iron-deficiency anemia, unless there is a concomitant cause of marrow hyperplasia, such as hemolytic anemias. Nonetheless, speculations on the cause of this condition in preh istoric populations abound, in part from a lack of medical docum entation in earlier times. Some of these suggested causes included sickle cell dis ease (Cooley and Lee 1925) and thalassemia (Angel 1978), with iron-deficiency anemia thought commonly to be the cause of this condition worldwide (Aufderheide and R odrguez-Martn 2005:349; Cohen 1989:107; Stuart-Macadam and Kent 1992:151-6; White 2000:394; White and Folkens 2005:320). To understand the process of iron-defici ency anemia, one must understand how the body stores and processes iron. Infants are born with adequate iron levels, which they
46 acquire from their mother in utero, even if the mother is iron deficient. However, breast milk is not iron-rich, and the growth demands of infants result in iron deficiency unless there is iron supplementation. In antiquity, th is may have been advantageous as the deficiency provides a resistance to infection; yet, today, with the pr esence of antibiotics, iron deficiency is not viewed as beneficial Lack of iron in the diet is the general explanation of iron deficiency in infants and children. In adul ts, it is usually the result of a gastrointestinal problem (e.g., parasites, ulcers, or cancer), pregnancy, abnormal menstrual bleeding, or customs. A combinati on of factors (disease and poor diet, or pregnancy and poor diet or dise ase) affect an individuals ir on status (Table 3.1) and may also create a drain on the bodys iron stores resulting in ir on-deficiency anemia (Cohen 1989:107; Holland and O'Brien 1997; Keita 200 3; Larsen 1997:30; Smith 1998; White 2000:394). It has been thought for some time that poro tic hyperostosis and cribra orbitalia are related to nutritional iron deficiencies (dietary model) (El-Majjar et al. 1975) rather than being adaptive responses to parasitic infec tion (parasite model) (Kent 1992:3; Wadsworth 1992) (see Hengen 1971 and Holland and O'Brien 1997 for a detailed discussion of these models). This is because interpretations were made primarily from the remains of humans who relied upon a maize-based diet in North America. In the Old World, the explanation of porotic hyperostosis resulting from a hemo lytic process, such as thalassemia, seems more plausible (Rothschild 2000). Stuart-M acadam (1986:284) notes the dietary model raises some questions when compared to data that show some individuals having anemia and others with the same diet not having anem ia, or anemia occurring in periods when an individual is vulnerable to disease (e.g., pregnancy and early childhood), and the fact that
47 Table 3.1. Factors affecting iron status (after Stuart-Macadam 1998:Table 4.5) Factors that affect iron absorption Children absorb more than adults Females absorb more than males Pregnant women absorb more than non-pregnant women (after the first trimester) Lactating women absorb more than non-lactating women Iron-deficient individuals absorb more than iron-replete individuals Cultural factors (smoking, alcohol consumption, the use of iron cookware, and the use of iron supplements) Diet Iron absorption is inhibited by: phylates (cereals, nuts, legumes), calcium (in the form of dairy products and supplements), polyphenols (plant metabolites found in tea, coffee, cocoa, red wine, vegetables and legumes) Iron absorption is enhanced by: ascorbic acid, consumption of meat products, and fermented foods Blood loss Parasites (e.g., hookworm, malaria, schistosomiasis) Gastrointestinal disorders (e.g., ulcers, hemorrhoids, colitis) Drug use (aspirin, antibiotics) Menstruation Iron withholding Hypoferremia as a defense to disease Anemia associated with infection Anemia associated with chronic disease Genetics Hemochromatosis Hemoglobinopathies porotic hyperostosis has also been demonstrated to occur in cases of genetic anemias (Stuart-Macadam 1987a). In addition, there is an increase in the cases of porotic hyperostosis and cribra orbitalia from the U pper Paleolithic through the Neolithic, with it
48 being more common around the equator and envi ronments with high levels of pathogens (such as sedentary, aggregated villages); most of the time these cases are related to a high pathogen load, rather than poor diet (K ent 1992:17; Stuart-Macadam 1992a). StuartMacadam (1992b) notes diet has been found to play actually a minor role in the development of iron deficiency anemia. Since research has indicated a strong positive to a strong negative correlation between porotic hyperostosis and infectious di sease (Kent 1992:3), fu rther interpretations have been made about these conditions being an adaptation to the environment, rather than evidence of an individuals i ll health (Stuart-Macadam 1991, 1992b). This relationship is possible because of the interac tion of iron levels with ones susceptibility to infectious disease (He ngen 1971; Klepinger 1992; Stuart Macadam 1992b; Weinberg 1992). Since iron is essential to the growth of many microorganisms and fungi, high iron levels can make an individual more prone to in fections. Therefore, it is also possible the presence of porotic hyperostosis indicates the organism is less susceptible to infection (Klepinger 1992:123-124), and its presence in a population may be an indication of an attempt to adapt to the environments pa thogen load (Stuart-Macadam 1992b). However, iron deficiency can be either beneficial or detrimental during an immune response (Dallman 1987:333). Strauss (1978) analyzed literature regarding iron deficiency, infections, and immune response and notes in some instances those with iron-deficiency anemia experience fewer infections, and there is a general depression of the inflammatory response. The ability of the host to wit hhold iron from invaders as a defensive mechanism, known as nutritional immunity, results in hyperfe rremia and can create more problems for individuals with infection because it makes them more susceptible to other
49 invading pathogens that rely on this elem ent (Weinberg 1974:955). Oppenheimer (2001) has studied the relationship between iron-defici ency anemia and infectious disease. The results of this study indicate iron therapy or supplementation has to be beneficial in some instances of infectious disease, such as respiratory illnesses (although multiple studies have produced conflicting results). Therefore, it is also possible th e etiology of porotic hyperostosis is a direct response to a diseas e or inflammation and not an adaptive process (Kent 1992:3). However, in cases of malari a, this type of malnutrition does not necessarily protect against the pathogen, but has been demonstrated to be a risk factor for it (Snow and Gilles 2002:101-102). Holland and OBrien (1997:190) stress the importance of including diet when examining cases of porotic hyperostosis and cribra orbitalia, and think porotic hyperostosis and cribra orbita lia result from a combinati on of factors more-or-less equally important. However, Kent (1986:628) ca utions attributing iron-deficiency anemia to dietary factors without consid ering that the other effects of sedentism, such as viruses, bacteria, and parasites, can lead to incorrect conclusions. In fact, it is not correct to assume dietary factors play even a small role in iron-deficiency anemia, especially when the results of research conducted under this uns ubstantiated premise (i .e., diet is the sole cause of iron deficiency anemia) are used to determine the funding of programs in developing countries. Therefore, when analyzin g human remains, it is important to be aware of both environmental and dietary condi tions. The evidence should be examined as completely as possible, considering a combin ation of factors, and incorporating multiple lines of evidence to make an educated interp retation of the causes of this condition (e.g., Holland and OBrien 1997:191; Martin et al 1985; Stuart-Macadam 1992a). Also, the
50 various causes of anemia could lead one to attribute the discrepancies between the health of individuals of different ages and sex as be ing related to access to dietary resources or status, when in reality, their dietary resources could be the same but their physiological needs could be different, causing some indi viduals to develop bony reactions, whereas others do not. Furthermore, histological exam inations of porotic hyperostosis and cribra orbitalia have identified different causes (e.g., anemia, inflammation, postmortem alteration) within the same population (Wapler et al. 2004). Research by Hengen (1971:71) shows the occurrence of porotic hyperostosis and cribra orbitalia increases closer to the equa tor, with high incidences in tropical and subtropical areas. It has been observed that severe anemia can occur in those infected with malaria parasites and c ontinue after the disease has resolved (Sri-Hidajati 2005). Although the disease processes that cause malarial anemia and related bone marrow suppression are poorly understood, it is po ssible these are an adaptive response to malaria, such as thalassemia major or minor or sickle cell anem ia (Klepinger 1992:122; Larsen 1997:30). Capasso and Di Tota (1995) attest that regarding malaria, the paleopathologic lesions are all related to porotic hyperostosis Their research indicates that a review of paleopa thologic data suggests P. falciparum became established in the Old World during the deforestation that acco mpanied the adoption of agriculture. Angel (1966, 1967, 1972, and 1978) examined the relationship between ecology, health, and disease in the eastern Mediterra nean, in particular linking poro tic hyperostosis, as well as paleodemographic results and stature changes, to thalassemia and malaria. Sallares and Bouwman (2004) believe using porotic hyperost osis from bones found in archaeological sites is a promising approach to resear ching malaria in prehistoric Greece.
51 Most recently, Walker et al. (2009) argue that iron-deficient anemia results in a condition of decreased red blood cell production, and thus cannot be responsible for porotic hyperostosis. Instead, the conditions ar e more likely related to megaloblastic and hemolytic anemias, the former involving the lack of vitamin B12 (which can be insufficient in the diet or resulting from malabsorption) a nd the latter involving the loss of blood cells through hemolysis (as observe d in malaria infections and inherited anemias). In the cases of hemolytic and me galoblastic anemias, red blood cell production is increased (Walker et al. 2009:112). In additi on, Walker et al. (2009) state that cribra orbitalia is not likely related to anemia directly, but is more likely related to other nutritional deficiencies that may or may not occur with anemia. 3.2.2 Skeletal Indicators for Thalassemia Angel (1967:379-381) noted porotic hyperost osis and cribra orbitalia are the results of the hyperactive nature of the bone marrow when it is faced with having to replace red blood cells with a shorter-tha n-normal life span. He hypothesized that hyperostosis in facial bones (e.g., maxillae, greater wings of the sphenoid, zygomatic bones, and orbits of the frontal bone), long bones, and ribs was the result of an anemia, most likely thalassemia or sickle-cell anem ia, by correlating the geographic distribution of malaria and these bony responses, along w ith changes in sea le vels and ecological conditions over time (Angel 1966, 1967, 1972, 1978). He also thought the cause of the anemia, or thalassemia, could be from a diffe rent source, not related to malaria. At the time of his research it was difficult to identify bone changes associated with this anemia because there were few comparative colle ctions known to belong to those with
52 thalassemia (including X-rays ). In addition, current popula tions benefit from medical care, which would prolong thei r life and possibly modify the natural bony responses one would be subjected to in antiquity. However, one early documentation of thalassemia included a notation of impacts to bone, such as hyperplasia and erythroblastic anaplasia of bone marrow and changes in the thickness and width of the zygomatic bones (Chini and Valeri 1949:995-1003). From radiographs of afflicted individuals, Chini and Valeri (1949:1004) describe diffuse osteoporotic lesi ons on the skulls of individuals suffering from this condition that can be in terpreted as porotic hyperostosis. Skeletal changes can occur in individual s with thalassemia during the first six months of life (Baker 1964; Soren et al. 1995). Thalassemia major produces the most severe skeletal changes, in cluding marrow hyperplasia. Long bones are noted to have an increase in Harris lines and flask-shaped deformities because of premature epiphyseal closure (especially the proximal humerus a nd distal femur) (Exarchou et al. 1984) and a widening of the medullary cavity. Severe chan ges in cortical bone, such as fissures, thinning, and mineralization def ects, also occur. Dwarfism, osteoporosis with fish-like vertebrae, pathological fractures, a honey-comb pattern of the bones in the hands and feet, with enlarged nutrient foramina of the phalanges (F ink et al. 1984), a hair-on-end appearance of cranial bones, an expansion of ribs with cortical destruction, and trabecular bone arranged diagonally with right-angle crossings have also been observed. Other problems with the bones of thalassemic indivi duals include deformities, scoliosis, and osteoporosis. Nerve compression and bone pain have also been reported by thalassemic patients. In general, skeletal growth is reta rded in thalassemic indi viduals (Amini et al. 2007:42; Aufderheide and Rodrguez-Martn 2005:347; Domrongkitchaiporn et al.
53 2003:1682; Dresner Pollak et al. 2000; Rioja et al. 1990:69; Singer and Vichinsky 1999). Unfortunately, quantitative studies of individu als with thalassemia major are rare and are usually limited to dental studies (see Amin i et al. 2007:37 and Bassimitci et al. 1996 for quantifiable studies), leaving qualitative de scriptions of the condition at best. These include the observance of bone marrow e xpansion in the skull and malocclusion. Thalassemias (usually beta-thalassemia) produ ce facial features th at are described as mongoloid or rodent facies (Amini et al 2007:37; Tayles 1996). The malocclusion is described as a protrusive premaxilla with flaring and spacing of the upper teeth, increased overjet and reduced overbite with other morphol ogical attributes including prominent zygomatic bones, a depression of the bridge of the nose and a partially obliterated maxillary sinus (Amini et al. 2007: 37). In the skull, there is also maxillary prognathism, pronounced vertical growth of the mandible, prominent mandibular incisors, and a narrow nasal cavity (Amini et al. 2007:42; Bassimitci et al. 1996:157). A study by Pusaksrikit et al. (1987) demonstrated that there is no indication of underbites in thalassemic patients. The minor form of thal assemia does not result in any osseous tissue changes according to Janssens (1981:104); howev er, others (as discussed in the previous section) think porotic hype rostosis and cribra orbitalia may be indicators. The oldest possible case of thalassemia, inte rpreted from gross skeletal analysis of remains from the submerged Neolithic site of Atlit Yam off the coast of Israel, dates to 6000 BC (Hershkovitz and Edelson 1991). Anothe r study of skeletal remains, from prehistoric central Thailand (c. 2000 BC), reports cases of possible thalassemia (in infants) that included skelet al responses such as cribra orbitalia, abnormally thick craniofacial bones, hypertrophy of the anterola teral sections of the frontal bones and the
54 zygomatic bones, and rounded edges of the orbital rim and inferior margins (Tayles 1996). Further recorded evidence of thalassemi a in older children included severe cribra orbitalia. Additional conditions observed by Ta yles included thin cortices and enlarged medullary cavities in long bones, hypertrophied metatarsal and phalange shafts with gross porosity of the cortical bone, and enlarged nutrient foramina on hand phalanges. The author notes while many other diseases can produce these bony res ponses, it is likely to her that these data, taken into considerati on with the age at death of the individuals studied, the environment, and other suggesti ons of prehistoric hemoglobinopathies, point to the possibility of one of the th alassemia syndromes (Tayles 1996:25). 3.2.3 Ancient DNA The examination of aDNA is a subdiscipline of molecular paleontology, and it involves the study of genetic materials obt ained exclusively from plant and animal remains (Wayne et al. 1999:458). Questions addressed by aDNA analysis include those concerning systematics, changes in genetic diversity, mutations in populations, migration, sex identification, kinship an alysis, and ecology or paleo ecology. The analysis of aDNA has provided some promising results useful for the reconstruction and understanding of various human issues, such as mummification processes, the spread of disease, and the effects of diets (Brown 2000:462-469; Rollo et al. 1999:111; Wayne et al. 1999:458). Often, this reconstruction is done by ex tracting aDNA from bone or mummified human tissue and amplifying it using the polym erase chain reaction (PCR) (Rollo et al. 1999:111). PCR is used to study aDNA b ecause it is a sensitive method, and identification can be accomplished with small amounts of mate rial, even single molecules
55 (Brown 2000:470; MacHugh et al. 2000; Pb o et al. 1989:9710); however, a minimum number of molecules per amplificati on should be used (100-1,000) to reduce contamination or PCR errors (Handt et al. 1996:375). PCR works by copying a defined sequence of DNA (template) many times, usi ng primers (two short pieces of DNA) to define the portion of the DNA to be copied. This method can only be used when the sequence of the DNA to be studied is known in advance. Brown (2000) provides a good summary of the nature of DNA and the PCR methods used for detecting ancient malarial DNA. The binding process of DNA to hydroxyapatite makes bone an ideal candidate for the extraction of aDNA (O'Rour ke et al. 2000:221-222), with microscopic preservation a good indicator of the ability to extract aDNA. Rib bones have been found to be the best candidates for aDNA extraction; not only doe s their spongy nature yield more DNA, but they are often of minimal pa leopathological importance and rarely missed from museum or archaeological collections (O'Rourke et al. 2000:222). Problems associated with the recovery of aDNA are preser vation, degradation, contamination, and organic inhibitors (e.g., tann ins, humic and fulvic acids) to the PCR process (O'Rourke et al. 2000:218-219). These issues make it a somewhat controversial method of analysis (Poinar and Stankiew icz 1999:8426-8430; Richards et al. 1995). For instance, the preservation of DNA is infl uenced by many environmental factors and is poorly understood. Favorable conditions for the preservation of aDNA are environments with low temperatures, high salt concentratio ns, and result in ra pid desiccation. Decay still occurs even in the most favorable e nvironments; however, degradation will occur at a faster rate by processes such as oxida tion, radiation, deamination, depurination, and
56 other hydrolytic processes (Hofreiter et al. 2001:353). In addition, the DNA of other organisms, such as bacteria and other mi crobials, may cause contamination (Machugh et al. 2000; Rollo et al. 1999:111). Further c ontamination may occur in the lab during extraction. Since modern DNA sequences are indistinguishable from ancient human DNA, the handling of remains by living humans introduces the possibility of contamination making interpretation even mo re difficult. Ancient DNA extracted from human remains is often rare, because fre quently little or no DNA survives and amplification by PCR is not possible (H ofreiter et al. 2001:354-355). One way to determine if there is enough DNA in a sample to proceed with analys is is through the use of amino acid analyses that isolate the type of amino acids found in DNA before they racemize (Poinar et al. 1996). Racemization is the process of the three-dimensional structure of an amino acid changing to a mi rror form over time (see Poinar et al. 1996 and Hofreiter et al. 2001:354 for more informati on about this process). Because of these challenges, rigorous protocols must be us ed to obtain uncontaminated samples and achieve accurate results. It is advisable to have at least two in dependent extractions (completed at different labs preferably), as well as cl oning of amplification products when any sequence heterogeneity occurs in direct sequence reactions. More experimentation with aDNA is recomme nded, with an emphasis being placed upon designing experiments that can be rep licated (Handt et al. 1994; 1996:375). Acute malaria may result in epidemics in which many individuals, especially children, die quickly. The results of this disease may not leave marks on bones; however, since P. falciparum affects the marrow of the bone, it may be possible to extract malaria DNA from human bones. An example of the use of aDNA to detect malaria in an
57 archaeological setting was the examination by Soren (2003) of a suspicious mid-fifth century burial of 47 infants at the site of Lugnano in Teverina, Umbria, Italy. This analysis ultimately revealed the presence of P. falciparum DNA. Soren (2003) used multiple lines of evidence to come to this conclusion, including analyzing the type of burial and the stratigra phy. He concluded it was likely th e result of an epidemic over a short period of time. Additional support from palynological evidence, the condition of the bones, types of tombs, ritual offerings, and a history of the re gion also supported the malaria hypothesis (Soren et al. 1995). Many of the skeletal remains from this study had evidence of porotic hyperostos is, cribra orbitalia, double-wa lled long bones, and enamel hypoplasias. The analysis of the aDNA from Lugna no (Soren 2003) was conducted by Sallares and Gomzi (2001:211), who note more suitable methods need to be developed for detecting ancient biomolecules from mala ria parasites. They acknowledge other methods, such as detecting genotypic evidence of beta-thalassemia, can be used to identify populations afflicted with malaria. In this instance, they found material from one of the individuals had a sequen ce that was 98% identical to P. falciparum which is strong enough to support the argument of its presence in this instance (Sallares and Gomzi 2001:203). Another study produced a less favorable re sult. Taylor et al. (1997) developed a method for detecting the DNA of the Plasmodium genus; however, when applying this method to two historic examples of indivi duals who are believed to have died from malaria, only one of them produced positive results (Rollo et al. 1999:111). Mummified remains dating to about 700 BC also produced negative results with DNA analysis, yet
58 the same mummy produced positive results with the use of immunological methods (Taylor et al. 1997); most recently an ex amination of DNA extracted from mummies from ancient Egypt (dating to 14001300 BC) suggest the presence of Plasmodium falciparum which may have played a role in th e death of King Tutankhamun (Hawass et al. 2010:646). Malaria may also be inferred by the presence of thalassemia. Since thalassemia is a genetic disease, aDNA analysis may be able to demonstrate its presence. An example of an attempt to identify thalassemia genetically was made by Yang (1997) on 1,900 yearold remains from the Italian archaeological si te of Isola Sacra (which is located on the coast near Rome, and the remains have also be en tested isotopically). While his research did not show genetic evidence of one of the mutations for this condition from the five individuals in his study, he c oncluded future advancements in aDNA analysis may be able to provide more insight into this and other conditions. 3.2.4 Immunology Since establishing that hemoglobin (or fr agments of hemoglobin) can be detected in human remains dating up to 4500 years ol d, it has been suggested immunological assays may provide information about the health of past populations (Ascenzi et al. 1985). Immunologic techniques, su ch as dipstick kits (e.g., Para Sight TM-F test Becton and Dickinson, USA; ICT Malaria P.f. IC T Diagnostics, Australia; OptiMAL, Flow Inc., USA) (Hnscheid 1999) are used to de tect antigens associated with malaria from circulating blood in living populations. Fo r example, the antigenic basis for the Para Sight TM-F test, which has been used on remains from archaeological contexts, is P. falciparum
59 histidine-rich protein 2 ( Pf HRP II). Sullivan (2002:146) notes this is a stable protein and uses the diagnosis of P. falciparum infection on mummified remains from Egypt (Miller et al. 1994) to support this argument. These kits have produced positive results when used on culturally mummified remains (Miller et al 1994; Schiff et al. 1993). An example of the use of immunological technique s to identify malaria in an archaeological context is the examination of mummified remains from Gebelen, Egypt (c. 3200 BC) using the Para Sight TMF test (Massa et al. 2000). Tissues (i.e., skin, muscle, bone, and teeth) from 50 individuals were examined, and 40% of the people tested positive for malaria. In the individuals who tested positive for malaria ( P. falciparum ), 92% also had porotic hyperostosis and cribra orbitalia. Sallares and Gomzi (2001:198-199) bri ng up several problems with the Para Sight TMF test. For example, there are high rates of false positive and false negative results. Also, they disagree with Sullivan (2002) regard ing the stability of this protein. They state it is not known how Pf HRP II degrades over time. So far, aDNA tests have failed to confirm the positive results produced by Para Sight TM-F tests; however, this could be expected when dealing with mummified remains as the chemicals used in the mummification process might degrade DNA. Othe r immunological disorders, such as the rheumatoid factor, may also produce positive results (Grobusch et al. 1999; Hnscheid 1999; Mishra et al. 1999). Nevertheless, because of the burden of malaria in the world, new, rapid, inexpensive means for diagnosing malaria are being produ ced. Little or no research with rapid tests is being conducted upon ancient rema ins; more work needs to be done before these types of test s can be reliable in diagnosin g malaria in past populations (Sallares and Gomzi 2001:199).
60 3.2.5 The Biomarker Hemozoin Hemozoin (HZ), also known as malarial pigment, is a waste product of the malaria parasite. Hemozoin is produced to minimize the accumulation of heme, which lyses cells, including the malaria parasite, released from hemoglobin as protein-free ferriprotoporphyrin IX (FP) (Orjih 2001:746-751; Sullivan Jr. 2002:137). Hemozoin is formed through biocrystallization, also referred to as biomineralization, which is defined as the deposition of low molecular weight inorganic materialswithin or outside the cells of living organisms (Hempelmann 2007: 673), and has also been synthesized by Haemoproteus sp. (avian), Schistosoma sp. and Reduviid bugs (in the Americas); however, biocrystals from Schistosoma sp. vary in size, shape, and structure (Sullivan Jr. 2002:136-140). Hemozoin has also been interpreted as playing a role in the inhibition of the production of red blood cells (erythr opoiesis) (Casals-Pasc ual et al. 2006). Heme accumulated as hemozoin is considered the biomarker for detecting malaria infections when using laser desorption mass spectrometry (LDMS), and it is described as insoluble, dissolving at pH 10-11 (Hem pelmann 2007:672; Nyunt et al. 2005:485; Sullivan 2002:138). Hemozoin has been detected in blood samples by LDMS in instances where the level of parasite s in the blood was too low to produce positive results by screening for parasitemia, that is, this procedure is more sensitive than diagnosis by microscopic examination (Nyunt et al. 2005; Sc holl et al. 2004). The examination of bone marrow for hemozoin has also been shown to be more effective for diagnosing malaria in instances of fevers of unknown origins when diagnosis by other means did not produce conclusive results (Mirdha, et al. 1999). Research has shown there is extensive hemozoin deposition in the spleen and bone marrow (e.g., Martiney et al. 2000:2265). In the
61 autopsies of patients with multiple malaria infections, the bone marrow appears black; children with severe malarial anemia also have this pigment present in their bone marrow. Malarial pigment also discolors the brain, lungs, liver, and spleen (Sullivan 2002:134). At present, there have been no studies reported where hemozoin isolation has been used upon human remains found in an ar chaeological context. However, because of its insoluble nature, it may prove to be more reliable than aDNA analysis when hemozoin isolation methods are developed for sa mples from archaeological contexts. 3.3 Chapter Summary The use of human skeletal remains in paleopathological studies has given us a better understanding of the lives of past popul ations. In prehistoric people, remains provide information about the unknown; in historic populations, remains can provide supplemental information, verification, or a be tter understanding of historic documents. In particular, there are five statements that can be made about de tecting malaria in bony tissue, which are applicable to this particular study: 1. if malaria results in hemolytic anemia, then it is likely porotic hyperostosis and cribra orbitalia will be observe d in populations with malaria (e.g., porotic hyperostosis and cribra orbita lia resulting from malarial anemia and found to be correlated to positiv e immunological test s by Massa et al. 2000 and observed in remains that ha d evidence of malarial aDNA by Soren 1995);
62 2. if a pattern of bony responses relate d to thalassemia is observed in a skeletal collection, then it is proba ble these individuals are from an environment with endemic malaria; 3. if the conditions are good for the pres ervation of the DNA, then it may be possible to extract malarial aDNA from the bony tissue of infected individuals; 4. if malaria was present, then antigen s associated with malaria may be detected in bone tissue us ing immunological assays; 5. if malaria results in the deposition of hemozoin in bone marrow, residual amounts may be detected using physi cal, chemical, or histological methods. Understanding how malaria affects a populati on and how its presence may be inferred from skeletal remains are the first two steps needed to test if malaria was present in a population. For this research, it is also n ecessary to understand the lifeways of the prehistoric and historic people of Sardinia, as well as the history a nd nature of malaria on the island.
63 Chapter 4: Sardinia Many factors must be addressed to understand the health of a population in prehistory. By examining the geography, clim ate, and environment, we can understand the physical surroundings in which a population lived, and in this case, if there are indications of suitable cond itions for the malaria disease vector and parasite. Furthermore, understanding the culture of a peopl e, from diet to trade, is necessary for identifying possible interactions that may have either aide d or hindered the transmission of malaria. Historic documents also provide inform ation about disease on Sardinia. From these, scholars have attempted to reconstruct the transmission of malaria to this island. This has resulted in the formation of unt ested hypotheses regard ing the presence of malaria on the island. By using methods similar to those of Angel (1964, 1966, 1967, 1972, 1978), Brown (1984) suggests a study can be undertaken to address the question: was malaria present in Sardinia during prehistory? 4.1 Geography, Climate, and Environment Sardinia (Figure 4.1), the second largest island in the Medite rranean, measures 24,000 km in area and is roughly rectangular. Ancient Greeks referred to Sardinia as Ichnussa or Sandaliotis, probably because they thought it was shaped like a sandal (Webster 1996:28). This island is located west of Italy and south of Corsica. Together
64 Figure 4.1. The location of Sar dinia and Corsica in relation to Sicily and the mainland (image by the SeaWiFS Project, NASA/Goddard Space Flight Center, and ORBIMAGE 2006) Corsica and Sardinia are known as the Sardo-Corsican Massif (sometimes referred to as the Corsico-Sardinian Massif) (Massoli-Novelli 1986:3). Even though these islands are close to each other, their geographic featur es vary. While Corsica is marked by sharp mountains, Sardinias landscape is more di verse, ranging from plains under 200 m in elevation to the highest point, Mon ti del Gennargentu, re aching 1834 m. Although Corsica had similar cultural developments as Sardinia, the island was too rugged and lacked the resources to sustain populations as large as those on Sardinia (Dyson and Rowland 2007:23). In general, there are three distinct zones in Sardinia. They are the lowland plains, the middle uplands, and mountainous regions. Th ese regions are defined by the locations identified in Figure 4.2.
65 Figure 4.2. Map of Sardinia with locations discussed identified (image by Jeff Schmaltz, MODIS Rapid Response Team, NASA/GSFC 2003) First, the lowland plains include thre e regions. One region, La Nurra and east Anglona, is in the north and stretches southeastward thr ough the valley of the river Mannu and southwest to the Coral Coast. In th e south, there are two lowland plain areas, the Campidano and the Sulcitano. The Camp idano is a broad, trough-like area extending for approximately 100 km from the Gulf of Oristano to the Gulf of Cagliari, and the Sulcitano is a smaller region between the Iglesiente uplands and the island of SantAntioco. These plains are low, arid, a nd have been used for agriculture throughout
66 history. For example, the Campidano Plain wa s used to produce grain for export to the mainland in Roman times (Guido 1963:23-29; Webster 1996:33-34). There are a few metalliferous ore deposits in these regions, specifically small copper deposits along the northwest coast at Argentiera, Alghero, and Montresta. In addition, the only known source of Sardinian obsidian, Monte Arci, is near the Gulf of Or istano and overlooks the Campidano Plain. Second, the middle uplands are thinly-woode d regions that run from northwest Anglona south through Logudoro, Marghine, and Arborea and continue through the southeast regions of Marmilla and Trexenta This landscape is extremely diverse and rugged; however, there is the benefit of more rainfall in the middle uplands, which made the region useful for producing grain and barley in Sardinia during the nineteenth century. In addition, copper and tin are f ound in this area (Guido 1963:23-29; Webster 1996:37-38J). The restrictive factors of this region are the rocky landscape and erosion that occurs along the slopi ng land (Webster 1996:37-38). Finally, the mountainous areas account for over half of the is lands landscape and include two chains that cove r two distinct zones. The la rger chain is contiguous and covers the eastern half of the island, while the smaller chain is in the southwest section of the island, with the Campidano Plain separating them. The Sulcitano Plain separates the smaller chain from the southwest coast. While the mountainous regions have more precipitation, both zones lack high-quality soil, and th e terrain makes communication difficult. These areas are notorious for persisti ng social conflicts that resulted in banditry, vendetta and livestock theft, as well as resolution through an informal organization of patrols, baracellato (Webster 1996:39), especially dur ing times of invasion by Roman
67 troops (Brown 1984:227). Nonetheless, today, th e valleys in these regions are used for cultivation, viniculture, and sheep, goat, a nd cattle husbandry. Beginning in antiquity, these mountainous zones have been a source of minerals, such as argentiferous galena, lead, and copper (Guido 1963:23-29; Webster 1996:38-40). The summers on Sardinia are typically dry and hot. The winters are cool and dry with strong winds, maestrale which can reach up to 120 km/hr and have contributed to the erosion of rocks, the formation of dunes, and bending of trees (Massoli-Novelli 1986:4). The rivers and streams are few in number (Coghinas, Tirso, Flumendosa, Mannu, and Cedrino), and there is only one fr eshwater lake, Lago di Baratz. Throughout the millennia, people have used other freshw ater sources on the island, including springs in the upland zones and rainwater (Webst er 1996: 31-32). Current populations have benefited from the construction of dams a nd artificial reservoirs (Vinelli 1926). Unfortunately, there are numerous gaps in the environmental and ecological data pertaining to Sardinian prehis tory. Webster (1996:42-43) note d most of the data relating to environmental conditions were taken from historical accounts and analogies from other areas, such as paleoenvironmental reconstructions from Corsi ca, and it is not certain if these data can be used to draw conclusions about prehis tory. Rowland (2001:2) also acknowledged little is known about the paleoclimate of Sardinia (see Madrau 2004). However, inferences have been made by an alyzing floral and faunal materials. For example, the environment on Sardinia can be inferred from research such as isotopic studies of shells and pollen an alysis. Research in these areas has demonstrated that the temperature and humidity reached their optimum extent during the Early Neolithic, 60005300 BC (Tykot 1994; Shackleton et al. 1984). This is interpreted as an indication of the
68 final trend in the full establishment of mixe d oak forests and evergreen species. The sea level continued to rise during this time. By about 5500 BC, it was between fifteen and ten meters below the present sea level, and by 4500 BC, it was six to seven meters below the present level. Other oxygen isotopic studies (see Whittle 1996), conducted in southern France and northern Italy, indicate a cooling trend around 3500 BC with a warming trend following at about 2500 BC. Pa lynological research by Reill e (1984), Renault-Miskovsky and Miskovsky (1969), and Vigne (1990:377) have shown there was significant deforestation at the end of the Neolithic. Vigne (1990, 1992) has outlined changes in ecology interpreted from the frequency of faunal remains found throughout Corsica and Sardinia. Not only do these changes explain ec ological variation, they also reflect the subsistence practices as humans se ttled on the Sardo-Corsican Massif. Zooarchaeological and fo ssil evidence support the hypothesis that humans began introducing species to Sardinia beginning in the Early Neolit hic, and this introduction resulted in the disappearance of endemic fauna (Vigne 1990:371). In fact, Vigne (1990, 1992) concludes that most, if not all, of the present animals on Sardinia are the result of human introduction to the isla nd. These species include fox ( Vulpes vulpes L.), cattle ( Bos taurus L.), dog ( Canis familiaris L.), and various other domestic animals ( Ovis aries L., Capra hircus L., Sus scrofa L.). Upon their introduc tion, these species caused ecological changes to the vege tation in the coastal and lowl and areas. Furthermore, an analysis of the amount of non-endemic animal s present on Sardinia, as well as research regarding sea currents and po ssible routes of travel, lead s Vigne (1990:377) to conclude there was secure, efficient sea transportation by the end of the sixth millennium BC;
69 however, it is likely that this was possibl e during the early Neolithic, as supported by archaeological evidence (e.g., number of early Ne olithic sites, types of domesticates, and obsidian distribution). Ecological changes interpreted from faunal analyses can be summarized in five stages (Vigne 1990, 1992). The first stage occurs in the Late Pleistocene when there is evidence of nine to ten taxa that existed in a stable state on Sardinia, with no changes in the Paleolithic. The second stage followed in the Mesolithic (eighth millennium BC) with hunters creating a disturbance in these numbers specifically a reducti on of the number of larger taxa. The third stage is the resulting st abilization of the smaller animals with a few large ones, and the more common presence of humans. At the start of the Neolithic, there is the introduction and increas e in the number of domestic species represented on the island with the growth slowing and leveling off in the Bronze Age. The final stage occurs during the Iron Age, in which there is a ne w rise in the number of species and the introduction of game faun a during the Middle Ages. Today, the flora and fauna on Sardinia ar e quite different than they were in prehistory. Since the middle of the 19th century AD, deforestation, overgrazing, and burning have resulted in smaller forests which exist at higher altitudes. These forests are composed of several species of oak ( Quercus robur, Quercus ilex, Quercus suber ), olive ( Alea europea ), tamarix ( Tamarix gallica ), wild fig ( Caprificus ), elm ( Ulmus procera ), white poplar ( Populus alba), laurel ( Laurus nobilis ), wild pear ( Pirus comuis ), elder ( Sambucus nigra), and smaller varieties such as scrub ( macchia ), hawthorne ( Prunus spinosa), and holly ( Ilex aquirolium ) (Muroni 1980). At the lo wer elevations, perennial bushes and annuals are common, such as wild rose, ivy, juniper, gorse, lentisk, rosemary,
70 privet, laurels, heather, blackberry, myrtle nettle, and fennel (Asole 1982). Cork oak ( Quercus suber ), which is used for its bark, is the most common lowland tree (Webster 1996). The mouflon sheep ( Ovis musimon), a domesticate gone feral (Schule 1993:408), is a faunal species of note. This protected animal is limited to the upland forests of Sardinia and Corsica. In the past, howeve r, they were far more numerous. The fallow deer ( Dama dama), was common into the 19th century AD when it was poached to extinction on the island, and then it was reintroduced in 1968. Popular foods during prehistory include a dwarfed sub-sp ecies of the European wild boar (Sus scrofa meridionalis ), the fox ( Vulpes vulpes ), wild pigeon ( Columba livia ), and rabbit ( Oryctalagus cuniculus ), and these are still hunted today. A large, wild hare ( Prolagus sardus ) was also used as food into the Iron Ag e, but it is now extinct (Webster 1996). Changes in ecosystems can be the result of natural or human factors, and these may affect the distribution of species, the lands cape, the environment, or the climate. For example, substantial anthropogenic modifi cations to the landscape by 4000 BC had resulted in geomorphological modifications, soil erosion, formation of deltas and alluvial deposits in valleys, and erosion of gul lies (Webster 1996:42). While changes in ecosystems may not actually determine human be havior, it is likely ch ange does act as a constraining factor (Trump 1983; Whittle 1996), which is important to acknowledge when analyzing human behavior. In fact, the uniqueness of island populations and the constraints upon the resources av ailable have led some scientis ts to consider islands as contained laboratories for investigating specifi c research questions (see Patton  for a detailed discussion of this topic).
71 4.2 Humans and Sardinia There has been relatively little radiocarbon dating done for the island of Sardinia. Kra (1998:5) noted there are just ove r 100 radiocarbon dates resulting from archaeological excavations as of 1998, and only a few from recent studies (see Lai 2008; Tykot 1994). Therefore, the unde rstanding of the chronology of Sardinia (Table 4.1) is not very clear, especially when examini ng problems of human development, cultural change, and environmental conditions in this island environment. In addition, it is evident there is variation in the in terpretation of the chronology and cultural categorization of Sardinia, especially concerning the Bronze Age, which is discussed later. Also, Table 4.1. The chronology of Sardinia (after Tykot 1994:129) Lower Middle Clactonian? > 150,000 BC Upper Paleolithic Upper 15,000 11,000 BC Mesolithic Grotta Corbeddu 11,000 6000 BC Su Carroppu 6000 5300 BC Early Filiestru Grotta Verde 5300 4700 BC Middle 4700 4000 BC Neolithic Late Bonu Ighinu ----(San Ciriaco)---Ozieri 4000 3200 BC Initial Sub-Ozieri Filigosa Abealzu 3200? 2700 BC Full Chalcolithic Final Monte Claro 2700? 2200? BC Early Bonnanaro A Beaker A Beaker B 2200 1900 BC Bonnanaro B 1900 1600 BC Middle Nuragic I 1600 1300 BC Late Nuragic II 1300 1150 BC Bronze Age Final Nuragic III 1150 850 BC Geometric 850 730 BC Orientalizing 730 580 BC Early Iron Age Archaic Phoenician Nuragic IV 580 510 BC Punic 510 238 BC Republican 238 1 BC Late Iron Age Roman Imperial Nuragic V 1 AD 476 AD
72 interpreting Sardinian archaeological sites is difficult because of the reuse of settlement sites, combined burials in tombs used fo r long periods, and conservative metalworking practices (Ridgeway 1979-1980:58). Separated from Corsica by only a 12 km ch annel, it would seem logical this island was populated by humans from the mainland via Elba and Corsica. However, there is little evidence this route was used initially, and still, today, th ere is less contact than what would be expected between Sardinia and Corsica, as demonstrated by the genetic variability of humans (Francalacci et al. 2003) Because of the natural harbors, bays, and other calm inlets around Oristano and Cagliari, it is likely the island was populated through the regions of these pres ent-day cities, rather than along the rugged east coast of the island (Webster 1996: 28). 4.2.1 Upper Paleolithic and Mesolithic Lower sea levels at about 20,000 years BP provided suitable conditions for humans to settle in Sardinia. The climate during this time was also conducive to vegetal growth and the creation of marshy areas beneficial for hunter-gatherers. At the end of the Pleistocene, temperatures increased and s ea levels rose creatin g more swamps and lagoons along the coast, which allowed more types of fauna and vegetation to be supported. An analysis of evidence from Co rbeddu Cave (Hofmeijer 1987), on the eastcentral side of Sardinia, indi cates humans were first presen t in Sardinia during the Upper Paleolithic. Remains include a human ja w fragment dating to 20,000 years BP, but cultural horizons produced date s only as old as 13,000 years BP. Archaeological evidence from the Mesolithic in Sardinia is also mini mal. Three sites, Grotta Su Coloru (near
73 Sassari), Corbeddu, and Filiestru, have yi elded dates to the seventh millennium BC (Hofmeijer 1987; Levine 1983; Wilkens 2004). 4.2.3 Early Neolithic During the Early Neolithic (ca. 60004700 BC), settlements in Sardinia and Corsica (or at least the ones id entified) were primarily restri cted to caves (e.g., Filiestru see Trump 1984) and rock shelters found near the coasts or in lessmountainous areas. In addition, there are some openair sites known, such as Ingur tosu-Arbus southeast of Montevecchio. Early Neolithic mortuary practices are not well documented (Rowland 2001:14). It is thought society duri ng the Early Neolithic was composed of small bands, perhaps extended families with an economy reliant upon herding, foraging, fishing, and some agriculture (grains and legumes) (Row land 2001:14). Evidence of subsistence in the Early Neolithic includes use-wear analysis conducted by Hurcombe (1992a:84-85) on obsidian artifacts, in which it was determined that fish and other animals were being processed at the site of Grotta Filiestru. In the Early Neolithic, tools were made from flint, quartz, rhyolite, and especially obsidian. Monte Arci obsidian is thought to have been discovered by indigenous populations and distributed inte rnally. Obsidian has been f ound (Tykot 1999) at all Early Neolithic sites in Sardinia, and assemblages include geometric microliths, scrapers, burins, and points. In addition, Sardinian obsidian has been found on mainland Europe indicating contact with outside populations; although it is not possible to reconstruct the exact trade route or exchange it was probably at a small s cale initially (Tykot 2002).
74 The Early Neolithic is divide d into the cultural phases of Cardial I, Cardial II, and Epicardial (or Filiestru) (Ta nda 1998a, 1998b). The pottery in Early Neolithic Sardinia is comprised of impressed wares (which ar e common throughout southeastern Italy, southern France, North Africa, and the Iberia n Peninsula), and it is found in the form of simple bowl and jar forms, usually with rounde d bases. This pottery is decorated with impressions made by cockle shell, or Cardium and woven mats (Rowland 2001:11). No chemical or thin-section analysis appears to have been done on this pottery from Sardinia to determine if they are of local origin (Tykot 1999:71). Dyson and Rowland (2007:2831) hypothesize that this ceramic technology an d the related production skills could have been exchanged for obsidian, or raids could ha ve brought potters to Sardinia from other areas. Obsidian could also have been trad ed for seeds, cultivation and agricultural technology, and imported animals (e.g., sheep, goat, cattle, and pig). 4.2.4 Middle Neolithic The Bonu Ighinu culture is associated with the Middle Neolithic (ca. 4700 4000 BC), with its height occurring at ca. 4500 BC (Rowland 2001:15; Dyson and Rowland 2007:19). Sites from this time are located in rock shelters and caves, such as Corbeddu, Filestru, Sa Ucca de su Tinti rriolu-Mara, and Grotta Rifugi o-Oliena. There are also many village settlements, which could have s upported small bands and extended families (Dyson and Rowland 2007:34; Trump 1989; Tykot 1999:72). In particular, village sites are predominant on the Campidano Plain, alt hough caves and rock shelters were also used in this region (de Lanfra nchi 1992). Open-air sites are found in the Sinis area, rock shelter and cave sites are found in the Iglesi ente region, and both open-air and cave sites
75 are found near Cagliari. Middle Neolithic cave sites in the Ca gliari region were probably used for sheep herding, and they were not oc cupied continuously (Michels et al. 1984; Tykot 1999:72). Some Bonu Ighinu s ites are located well into th e interior of the island (e.g., Grotta Pitzu, Polu Meano Sardo). One example, the site of Cuccuru SArriu (Santoni 1989), shows evidence of reed and timber huts, as well as rock-cut tomb prototypes, known as tombe a forno (Rowland 2001:15). The burials at this site included offerings such as pottery, lithics, bones, stone bracelets, armlets, red ochre, and obese stone female statuettes, with one person in an individual burial clutching one of these figurines (Rowland 2001:15-16). This is the firs t evidence of an ideological system in Sardinia and of the possible emergence of lo w-level hierarchies, probably resulting from trade (Dyson and Rowland 2007:34-35). A few sites studies of Middle Neolithic sites have produced evidence of cereal processing and increasing cultivation, includ ing the presence of axes and grinding tools in the archaeological record, the clearing of fo rests, and the remains of barley, grain, and legumes. It has been suggested crop rotation was occurring during this time, and grinding stones were used to process olives. While sh eep and goats were still a primary source of meat, cattle and pigs become less important a nd shellfish and land snails became a part of the diet. Hunting and gathering were also methods still used to obtain food (Rowland 2001:15). The archaeological record from the Middle Neolithic in Sardinia shows a considerable increase in the use of obsidian from Monte Arci to manufacture tools (Hurcombe 1992a, 1992b; Hurcombe and Phil lips 1998; Tykot 1999). In addition, one type of pottery from this period, Bonu Ighinu, which is homogeneous throughout the
76 island, was made with more detailed craftwor k and decoration than that of the Impressed Wares (Tykot 1999:72). This pottery, with punc hed and incised decorative elements, is similar to that found on the mainland, the Balkans, and Sicily (Dyson and Rowland 2007:34). Another ceramic type from this peri od has been recently identified, San Ciriaco (Molinari 2002), which have a monochromatic slip (Rowland 2001:16).During this time, other items such as beads, greenstone axes and stone rings were also used (Tykot 1999:73). 4.2.5 Late Neolithic During the Middle and Late Neolithic, human occupation expanded throughout most of the coastal lowland areas on the west ern side of the island and to a lesser extent into the interior valleys (W ebster 1996:47). From the numbe r of archaeological sites found, it appears the Late Neolithic is the most active time before the Bronze Age (Webster 1996:47). However, this could be because earlier sites are less obtrusive or more fragile (Whittle 1988). The Late Neolithic (ca. 4000 3200 BC) is characterized by one cultural tradition called the Ozieri (a.k.a. San Michele), which was named after the cave of San Michele at Ozieri in north-central Sardinia. As of the start of the 1990s, there were three times as many sites identified from this period than from the Middle Neolithic (Atzeni 1981; Tykot 1999:73). Limited radiocarbon dates i ndicate this cultur e extends through the fourth millennium BC with a late or sub-Oz ieri tradition beginning in the early third millennium BC (Tykot 1999:73). Ozieri sites ar e found throughout all of Sardinia, in all ecological niches, and are mostly open-air si tes or located in na tural caves (Dyson and
77 Rowland 2007:36). The sites are unwalled, thus demonstrating a lack of defensive architecture. It is estimated many of the Ozieri sites were the result of population expansion, with only a few showing an earlier component (Dyson and Rowland 2007: 36). Catchment areas at these sites have a radius of about two km (Rowland 2001:17). The best-known settlement from this peri od is San Gemiliano-Sestu (Atzeni 1959-61). Located near the Gulf of Cagliari and a la goon, this site is about 100 m above sea level, faces a well-watered plain, and covers about 200-220 m2. San Gemiliano-Sestu was a village with at least 60 huts that were about three meters in diameter. The inhabitants subsistence methods included hunting and gath ering as well as agriculture (Dyson and Rowland 2007:36-37). Other Ozieri village si tes include Cuccuru SArriu and Su Coddu (Ugas et al. 1985), the largest known community. In addition to the settlements, island-wide arch itecture is also found in the forms of tombs and structures fo r communal activities (Lazrus 1999:126-128). The tombs of this time are domus de janas or rock-cut hypogea (Figure 4.3). Contu (1964) notes a similarity in the domus de janas and tombs found on the isla nds of Malta and Sicily. There are about 2500 of them known on Sardin ia, and well-known ones include Anghelu Ruju, Santu Pedru, Mannu-Porto Torres, S. Andrea Priu-Bonorva, and San BenedettoIglesias. These tombs have been reused through the Middle Ages; some of the burial chambers appear to reproduce living spaces in the home. Bas relief decorations of bull horns and geometric patterns were found in these tombs (Contu 1966; Rowland 2001:22). Dolmens, alle couvertes, and menhirs also are typical during this time (Rowland 2001:19-20).
78 Figure 4.3. Exterior (above) and interior (b elow) of the hypogea at Anghelu Ruiu 3rd millennium BC (images courtesy of Stephanie Del Rosario)
79 The most remarkable Ozieri site is Monte dAccoddi (T in 1992), which is located in Sassari. This site is a platform -type mound with a larg e altar and unexplained egg-shaped piece of sandstone near it (Rowland 2001:22). It is presumably a religious structure measuring 37.5 x 30.5 m with a heig ht of about 5.5 m. A 7 x 16 m rectangular structure sits on top. A 41.5 m long ramp allowed for access to this structure. It was originally a Bonu Ighinu villag e site, then an Ozieri village, which was replaced by an early Ozieri shrine, and finally with Mont e dAccoddi (Figure 4.4) (see Contu 2000). The size and complexity of this s ite demonstrates the existence of an elite recognized by many communities, while the absence of fortificati on probably indicates social control and lack of endemic warfare (Dyson and Rowland 2007:41). During the Late Neolithic, there is evidence of the accumulation of wealth in the form of storage pits; however, these storag e pits are only found at one site, Su Coddu (Ugas et al. 1985). Data that ha ve been recovered from this time indicate the use of crop rotation, supplemented with fishing, shellfish collecting, stoc k raising, and hunting. Although floral remains are rarely found at Late Neolithic sites, some recovered remains include durum wheat, einkorn, emmer, barley, lentils, fava beans, and peas (Lazrus 1999:126; Piga and Porcu 1990). In addition, m ills and grinders have been recovered. Faunal remains include bones of deer, boar, mufflon, cattle, swine, and sheep. Rowland (2001:18) noted that porotic hyperostosis as observed by German (1995a, 1995b, 1998), is found in human bones from this time, and was interpreted as result ing from a lack of folic acid, vitamin B-6, and iron. The Ozieri culture is also technolog ically homogeneous throughout Sardinia. Rowland (2001:17) notes this is a time of an increase in prosperity. Obsidian is still being
80 Figure 4.4. Monte dAccoddi (image courtesy of Robert Tykot) exported and is found at sites in the form of points, blades, scrapers, burins, and awls (Rowland 2001:17-18). Ozieri ceram ics are heavily decorated with stylized figurative motifs, such as zigzags, spirals, triangles, festoons, circles, and human figures that are impressed or incised on the pottery. These cer amics are colored red or white, and include new forms, such as bowls and cups with carinated rims, vases with tunnel handles, tripods, and amphorae (Lazrus 1999:127-128; Tykot 1999:74). The use of copper and silver, as demonstrated by objects found at Su Coddu-Selargius (Ugas et al. 1985), also starts during this time in Sardinia. Alt hough few in number, metal objects are found in the form of tools, weapons, and tripods (Lo Schiavo 1989b:231). The Ozieri culture may extend into the Chalcolithic. However, some scholars have identified this extension as a sub-Ozieri phase (Lazrus 1999; Santoni 1989; Tykot 1994).
81 4.2.6 Chalcolithic The Chalcolithic period in Sardinia, a millennium long transition (3200-2200 BC) between the Late Neolithic and the Bronze Age. It is not understood very well but includes the sub-Ozieri, Monte Clar o, and Beaker cultures (Knapp 1992; Tykot 1999:75; Webster 1996:52). Although some interpret thes e as individual cultures, others, such as Rowland (2001:29), believe they could be regi onal variations of a single post-Ozieri subculture. While the Neolithic in Sardinia is thought to have been represented by peaceful, egalitarian farmers with a predominantly agricultural econom y (Lilliu 1988), the Chalcolithic is viewed differently. The beginni ng of the widespread use of metals in the Chalcolithic and the presence of sites enclosed by megalithic walls could indicate this was a period of differential access to resour ces (perhaps from an increasing population). These walls were either used to protect econom ic interests or resulted from increasing social tensions between groups (Lewthwa ite 1986; Webster 1990, 1996). Also, with the first indications of defensive architecture ap pearing during this time, other suggestion of conflict is shown by daggers represented on menhirs and found in deposits (Rowland 2001:29). The movement into sites with evidence of enclosure for husbandry and cultivation purposes is thought (in part) to be a result of a cha nge in agricultural practices, from specialized to mixed farming (Blake 1999:42). The sites from this period occur mostly from the Gulf of Oristano southward and are often found on elevated posit ions overlooking valleys used for agriculture, access to waterways, and springs. This possibly indica tes an increase in competition (Bagella 1998). There is some reuse of Ozieri sites. Examples of Monte Clar o sites include Monte
82 Baranta-Olmedo and Biriai. A quarter of the Monte Claro sites are in caves, and they were typically used for burials and seasonal habitations, and are clos e to the subsequent Nuragic settlements. Domus de janas were still used, as were cist graves that appear to be a precursor to giants tombs, or tombe dei giganti (Dyson and Rowland 2007:47-51). Culturally, there was a high degree of continuity from the Late Neolithic to the Chalcolithic. Lithic industries showed little change, and th ere were few differences in ceramics which were classified as Abealzu and Filigosa (Lilliu 1988), although these were possibly just a variation of Ozieri (Dyson and Rowland 2007:44). The presence of Bell Beakers, biconical burial vessels, found in domus de janas during the end of the Chalcolithic has been interp reted by some as evidence of an intrusive group coexisting peacefully with indigenous Sardinians (B almuth 1992:676), but these also could be circulated prestige it ems (Rowland 2001:31-33). Copper has been found at Ozieri sites, and its use is contemporaneous with that on mainland Italy (Lo Schiavo 1989a; Lo Schi avo et al. 1985; Stech 1989). During the Chalcolithic, lead was also used for the re pair of pottery, weight s, and waterproofing (Atzeni et al. 1991). Metal pins, daggers, and swords were also used during the Chalcolithic, and loom weights, indicative of wool working, are found from this time (Rowland 2001:29-34). The Bonnanaro phase follows the Monte Cl aro culture, and it is defined by an undecorated ceramic type. Lilliu (1988:276-316) cl assifies this as Nuragic Phase I. This pottery is found mostly in funerary deposits from 2200-1600 BC (Rowland 2001:33-34; Tykot 1994).
83 4.2.7 The Bronze Age In Sardinia, the Bronze Age (specifically the Middle and Late Bronze Age) is known for the stone towers, nuraghi (Figure 4.5), which are found throughout the island, from coast to coast, in various topographic settings. It is from these towers that the Nuragic culture is named. There are appr oximately 7000 known nuraghi, whose function is still debated (Lazrus 1999: 129-130; Lilliu 2005). In genera l, nuraghi range in height from one to three stories and have internal sp iral staircases and corbelled vaults. Webster (1991) calculated the amount of time required to construct a nuraghe from ethnographic analogies, to be about 40 days. Trump ( 1992) summarizes the classification of the nuraghi into three cat egories: proto nuraghi (square and oval in plan with only narrow passages or corridors), classic nuraghi (single-tower, possibly used for defensive purposes), and complex nuraghi (probably used for defensive purposes). These categories are also used to divide the Nuragic pha ses (Webster 1996). Throughout the Iron Age, these structures were expanded upon with the addition of more towers. The distribution of the nuraghi across the island has also led to speculation about terri tories, ethnicity, and the relationship between boundaries and res ources, especially regarding competition, both external, and more so, internal (Bon zani 1992:210; Webster and Webster 1998:383). Because of the variation of types, the dist ribution of nuraghi am ong different ecological niches, and their large numbers, it is not likely they were used at the same time, which makes the analysis of land use much more difficult (until contemporaneous towers are identified) (Lazrus 1999:130). The Bronze Age in Sardinia is usually di vided into three periods (300 years each) from 1800-900 BC; in fact, it has been pointed out by Lilliu (1982) that it is better
84 Figure 4.5. The Su Nuraxi Barumini Compl ex (image courtesy of Robert Tykot) to consider this period in Nuragic divisions. An alternative, more recent chronology created from the results of absolute dating methods, considers the Early Bronze Age as ranging from 2200-1900 BC, the Middle Bronz e Age from 1900-1300 BC, and the Late Bronze Age as dating from 1300-850 BC (Tykot 1994). For the purposes of this dissertation, the more recent chronology will be used. While the Nuragic period is typically t hought to have begun with the appearance of the nuraghi themselves, other cultural materials, such as the aforementioned Bonnanaro ceramics found in mortuary rema ins, indicate that the transition to a homogenous culture may have started earlier. Th e presence of these ceramics in giants tombs indicates it is likely they predate the nuraghi and may be related to pre-Nuragic sites that are no longer present. This hypothesis is supported by the fact that these tombs were constructed in a manner using materials consistent with pre-Nuragic architecture
85 (Blake 1999:42-43). Bonnanaro ceramics have also been found in other burial types predating the giants tombs, such as natura l caves and rock-cut tombs, suggesting the mortuary practices were varied during the transitional time before the Nuragic phase. The Bronze Age is often presented as a tim e of warrior-pastorali sts who lived in a stratified society and were becoming in creasingly dependent upon specialized pastoralism. However, the ar chaeological evidence does not indicate an increase in the production of textiles, dairy products, or special ized structures for animals, but rather a mixed agro-pastoral economy that was a succ essful adaptation maintained through future periods. The location of settlements also s upports the argument for a mixed agro-pastoral economy and contradicts the picture of pure Nuragic pastoralism (Lazrus 1999; Rowland 2001:38). It appears to Lazrus (1999) that the utilization of diversified subsistence and economic systems including farming, sma ll-scale animal husbandry, hunting, fishing, gathering, trade, and mining during the Neolit hic and Bronze Ages, allowed the societies to remain extremely stable. In addition, the faunal evidence presented by Vigne (1990, 1992) does not support a change in subsiste nce methods. More re cent research by Lai (2008), involving stable isotope analysis to interpret diet, al so does not indicate a change to a pastoral way of life. In fact, Lazrus (1999) stated there does not seem to be any archaeological indications of change in social structure unti l the very Late Bronze Age or the Iron Age. During the Bronze Age, lithic tools were still being used. They include sickle blades, pestles, hammers, grinding stones, and weapons (such as the doughnut-shaped testa di mazza). Use-wear analysis on obsidian artifac ts from the Bronze Age site of Ortu Cmidu indicates obsidian lunates were used for the processing of plant material
86 (Hurcombe 1992b). Residues found on pottery from this period show the use of olive oil, wine, grains, barley, grapes, and almonds; also porcupines, birds, and hare also supplemented the diet (Rowland 2001:41). Cork was also widely used during this time. Although edible olives were being grown on Sardinia from the Neolithic on, there is evidence of olive oil being im ported from the Eastern Mediterranean during the Late Bronze Age, with mastic oil (from Pistacia lentiscus ) likely to be the primary type of oil used during the Nuragic Age (Bafico and Garibaldi 1998). Bronze becomes increasingly importa nt around 1200 BC with artifact assemblages including swords, daggers, spearheads, and statues (portraying boats, animals, carts, and people) (Lazrus 1999: 129; Lo Schiavo 1989b:245). Proof of this includes stone molds (monovalve and bivalve). It seems likely metallurg ical skills of this nature were the result of a Cypriot influen ce, which started with the importation and the subsequent imitation of the imported Cypr iotic goods (Lo Schiavo 1989a). Although its appearance in Sardinia is not certain, the lost-wax technique, creating a mold around a wax model that is subsequently melted allowi ng metal to be cast in the mold (which was known in the Aegean and Near East from th e Early Bronze Age), was also used (Gallin and Tykot 1993). Early Bronze Age technology consists of Bonnanaro pottery (2300-1800 BC), metal awls, pins, and knife blades. In the Midd le Bronze Age, the pottery styles included punctuated and combed wares, pre-geometric, and fine grey pottery. Metal tools are not found as frequently as they are at the Earl y Bronze Age sites, but dagger blades, flanged axes, and jewelry have been recovered. Th e technology in the Late Bronze Age includes the ox-drawn plow, and a great increase in me tallurgy with the inclusion of votive objects
87 and a few iron pieces (Webster 1996:66-108). There is also indication of trade from the east in the form of Aegean (M ycenaean) pottery and Cypriot copper ingots, with evidence of gift-exchange between Sardinian and the Aegean occurring by the end of the Middle Bronze Age (Knapp and Cherry 1994:146-151). Th e presence of copper oxhide ingots in Sardinia during the Late Bronze Age is often linked with Mycenaean trade. It has been noted (Lo Schiavo 1989a; Lo Schiavo et al. 1985; Stech 1989) that these ingots were not used for the manufacturing of items. Instead, isotopic analysis (Stos-Gale and Gale 1992) suggests local sources of copper were exploi ted for functional purposes, and the Cypriot ingots served some other function, such as prestige items or currency (Patton 1996:173). From the Late Bronze Age to the Iron Age, th ere is evidence of ritual activities in the form of sacred well temples and reunion hut s, which were communal gathering places; stone furnished cult sites; and nuraghi m odels made of stone or bronze after the construction of the nuraghi (Blake 1997:152-153). Overall, Webster (1996:111-125) describe s the settlement patterns during the Bronze Age progressing from occupation of the plains and middle uplan ds to habitation in all regions of the islan d. During the Early Bronze Age, the Campidano Plain and the middle uplands on the western side of the island were settled. However, it has also been noted the Campidano probably was not used prehistorically because land and water sources would not support the crops farmed then; also, the environment may have been unhealthy (Rowland 2001:37). Stru ctures at the beginning of the Middle Bronze Age include corridor/gallery nuraghi (Fig ure 4.6), also called pseudo-nuraghi or protonuraghi. These were built on raised platforms with stone-lined corridors. Staircases were used to access the upper level, where there were huts with reed or wooden roofs, from the
88 corridors. The construction of proto-nuraghi indicates the emergence of local and regional elites. Settlements at this time were usua lly open-air sites, cave sites, and protonuraghi, which are found most frequently in the intermediate level highlands. However, chieftains, dominant families, and large monumental complexes were present, which are indicative of an amassing of considerab le wealth (Dyson and Rowland 2007:62). True nuraghi begin to appear during the Middle Bronze Age, and there is a decrease in open-air and cave si tes. The dispersion of the nura ghi at this time may be the result of social fissioning, and these structures may repres ent fortified, nuclear family farmsteads. Perra (1997) interprets the nuraghi as being prestige goods, with the ability to construct one representing a su rplus of capital. During the Late Bronze Age, there is an enlargement of the nuraghi with three types of class settlements emerging. Class I settlements are hamlets and homesteads, wh ich are found in all regions of the island. Figure 4.6. Nuraghe Losa is an example of a proto-nuraghe It is located in centralwestern Sardinia (image cour tesy of Robert Tykot).
89 Class II settlements include complex mu lti-tower structures and are also found throughout the island. Class III settlements ar e fewer in number, although there are still many, and are comprised of heavy, multi-towere d antemurals (an outwork of high, strong walls used for defense) around bastions (a stronghold where people could go for shelter), with evidence of meeting rooms. The best exam ple of this is Su Nuraxi-Barumini (Lilliu 1967), which was being constructed from the mi d-second millennium BC to the eighth to seventh century BC and overthrown by the Carthaginians in the sixth century BC (Webster 1996:162-164). Other examples includ e Nuraghe Santu Antine-Torralba (Contu 1988) and Nuraghe Losa-Abbasanta (Taramelli 1 916). There is also an increase of openaired settlements without nuraghi during this time, especially in the central highlands of Barbagia, Nucrese, Goceano, Baronie, a nd the lowland Campidano Plain (Webster 1996:111-125). These open-aired villages had ac tivities such as w ool processing, cheese making, growing/collecting grains barley, grapes, almonds, olives, and stock raising and hunting (Dyson and Rowland 2007:70-71). The populat ion at the end of the Late Bronze Age was estimated to be 450,000 to 600,000 (Usai 1995:257). Additional Sardinian architectural stru ctures of the Bronze Age include well temples (Figure 4.7) ( tempio a pozzo) quarries, and various mort uary structures (i.e., gallery graves, cave tombs, and large free-st anding tombs). Giants tombs (slab-lined, rectangular funerary chambers, typically ha ving a semicircular forecourt) and sacred wells (representing a water cu lt) are found during this time specifically. The tombs are fewer in number than the nuraghi and likely served multiple villages or multi-farmstead communities (Dyson and Rowland 2007:82-83). We bster (1996:104) notes a spatial study might provide information about ritual geograp hy. The distribution of these features may
90 also be indicative of regiona l polities, with smaller nuraghi acting as channels between larger neighboring polities (Knapp 1992). Dolmenic giants tombs (Fi gure 4.8) were used before the construction of conical nuraghi during the Middle Bronze Age, and c ontinued to be used throughout this period Figure 4.7. Well temple at Santa Cristi na di Paulilatino (photo by author 2005)
91 for communal, secondary burials. Because of il licit activities and the reuse of tombs over time (Blake 2001:151), it is difficult to estimate the number of individuals interred in a tomb; however, it is likely that their use was restricted to elites or specific groups. During the Middle Bronze Age (1600-1300 BC), howev er, these tombs were constructed differently. Instead of slabs, these tombs were built with block-shaped stones and had walls replacing the stele. The entrances were stele or made of pierced stone walls in a similar fashion to those of the nuraghi. Most of these tombs had a single chamber, and sometimes they were continued constructi ons from older tombs (Dyson and Rowland 2007:79). While the chambers of the tombs were small, the semi-circular forecourts (sometimes with benches around the entrance) of the giants tombs were larger than the antechambers of the domus de janas and allowed for more complex rituals, which most likely involved families and clans (Rowland 2001:45). In addition, some tombs have menhirs and smaller standing stones known as betyls, or betili Intact skeletal remains and funerary deposits are found rarely in gi ants tombs, and when they are found the remains are often highly fragmented. The i ndividuals buried in these tombs are most likely elites. However, giants tombs were not found everywhere throughout Sardinia, and domus de janas caves, and rock-cut tombs conti nued to be used in the Nuragic period (Dyson and Rowland 2007:82-83). In the Late Bronze Age (1300/1200-900 BC), the giants tombs were made from well-cut as hlar masonry and share stylistic attributes of the sacred well temples of this period (Blake 1999:48). From about 900-500 BC (Early Iron Age), both the tombs and nuraghi becam e differentiated, perhaps representing the establishment of diversified identities, varying site function, and a reflection of intragroup competition, rather than the ne ed for unification (Blake 1999:48-50).
92 Figure 4.8. S Ena e Thomes, a giants tomb, loca ted in Northern Sardinia (image courtesy of Robert Tykot) The spatial orientation of tombs in rela tion to settlements suggests Nuragic tombs were shared, much as other aspects of peopl es lives, as indicated by living and working areas. Because the Nuragic period was less sta ndardized archaeologically, the location of the tombs served as a means to unify people who were dispersed over a greater distance. The design of the tombs, in conjunction with their location, probably acted as cultural markers for those communities (Blake 2001:159). The use of the various types of tombs may have overlapped from one Bronze Age period to another (Webster 1996:22). For example, Blake (1999:43) notes only a few giants tombs associated with the Nuragic culture have produced any material remains, and they are usually associated with the Bonnanaro B phase. Thus these tombs may ha ve been used earlier than the Nuragic phase, and some continued to be used thr ough the Roman Imperial period (e.g., Brunceu
93 Espis di Fontanazzu and Su Monte de s Ape-Loiri) (Rowland 1981:14,58; Pulacchini 1998:43). The sacred wells, which were enclosed underground, are anot her indicator of ritual activity. These wells had entryways si milar to those of giants tombs (Dyson and Rowland 2007:86). Votive offerings, at sites such as Abini-Teti ( early Bonnanaro) and Sos Malavidos-Orani (late Bonnanaro), indi cate the water cult date s as early as 2000 BC (Pais 1884; Taramelli 1931). Luxury goods in the votive offerings are considered to be evidence of elite patronage. The well at Santa Cristina-Paulilatino (Figure 4.7) had Phoenician and Punic imports and bronze and terracotta statues related to the Cult of Ceres and Demeter (Atenzi 1977; Spano 1857; Taramelli 1910; Webster 1996:184-188). In addition to sacred wells, Nuragi c populations had a variety of watermanagement practices. Some examples include the use of channel sy stems, fountains, and other wells. These also incl uded possibly using the nuraghi themselves to store water (Balmuth 1992:677). The site of Nurdle, wh ich was a Monte Claro site upon which a Nuragic site was subsequently built. At Nurdle, the nuraghe and the sacred well were combined in one structure, and the site incl udes an architectural feature of a fountain (Fadda and Madau 1991), which also had a rese rvoir to collect and store water from the fountain. A drainage system, comprised of ch annels and a bell-shaped structure that intersected a channel and was probably used as a well, at Nuraghe Arrubiu demonstrates another method for the collection and contai nment of water (Lo Schiavo 1990). Systems of channels have also been found at the site of SantAnastasia (Ugas 1990). Furthermore, the inclusion of wells in the central courts of Nuragic complexes has also been observed, such as those at Barumini and Madau.
94 Evidence of more ritual activity is found at Santa Vittoria-Serri (Puddu 1992; Taramelli 1931). This site has data supporting a variety of cult functions related to animal fertility and social and political cohesion. Fo r example, there are streets at this site connecting the well to other buildings, such as an altar, a possible meeting hut, the supposed chiefs dwelling, and a festival hall. Votives were found from Proto-Villanovan and Etruscan Italy, as well as Punic material indicating the interaction of the Nuragic people with the wider Medite rranean (Dyson and Rowland 2007:89). Furthermore, there is additional evidence of contact with Etru ria, through the importa tion of bronze fibulas from the Central Italian mainland, and the finds of bronzetti in Etruscan tombs (Ridgeway 1979-1980:61). There is also ev idence (an anthropometric sarcophagus) suggesting contact with or settlers from the Levant around 1100 BC (Bartolini 1997). 4.2.8 The Nuragic Periods during the Iron Age During the late eighth century BC, Phoe nicians were visiting Sardinia and establishing more permanent ports (Barreca 1989:210). In some instances, Phoenician settlements were constructed on existing or abandoned Nuragic sites, first along the coast and then inland. However, there was a lack of Phoenician settlements on the east coast of Sardinia, yet trade between Nuragic populati ons and the Etruscans was still important there during the sixth and fifth centuri es BC (Lo Schiavo 1996; Tronchetti 2000). The first Phoenician colony founded was Nora, on the southern coast of the island near Cagliari. The Nora Stone, which was found at this site, is a tablet with a Phoenician inscription. Interpretations of this stone vary and include the documentation of a Phoenician victory over the Nu ragic people when an attempt was made to take mining
95 and industrial resources (Ridge way 1979-1980:59), a Phoenician victory in the Battle of Tarshish (Cross 1986), or refuge of the Phoeni cians at Nora after they were driven from Tarshish by a battle or storm (Peckham 1972). The relationship between the indigenous populations and the Phoenicians is not well understood. The excavation of archaeologica l sites, such as Monte Sirai-Carbonia (Moscati 1993), have uncovered a mixture of Nuragic and Phoe nician artifacts, which has been interpreted as an example of peaceful interactions; however, it may also be indicative of trade or looting (Moravetti 1992). Dyson and Rowland (2007:111) liken the relationship between the Nuragic people and th e Phoenicians to that of Native Americans and the French settlers of North America. After the middle of the sixth century BC, Carthaginian expeditions to Sardinia occurred in response to the Greek colonization of the northwestern Mediterranean. Initial attempts to colonize Sardinia failed, but they later succeeded, although they were met with resistance from both indigenous popul ations and Phoenicians (Lilliu 1992). Carthaginians attacked the coastal center s first, to prevent others from accessing resources, such as grain, mi nerals, and humans. Movement in to the interior of the island resulted in the destruction of Nuragic settlements and the development of the latifundia (large estates used for agriculture, whic h were usually dependent upon slave labor) system (Brizzi 1989; Marasco 1988) and the cons truction of border fort ifications in the fifth century. The construction of these fortif ications has been thought to have kept the Nuragic populations under c ontrol (Barreca 1978, 1986); however, another interpretation (Lilliu 1988) suggests this was unlikely because the Carthaginians lacked the
96 organization and leadership to enforce this system, and the use of the fortifications represents an adoption of technology and acculturation to Punic lifeways. Although the Carthaginians were interest ed in developing a trade network and utilizing the island for its agricultural pot ential, the Nuragic populations still had maintained their own independent trade netw ork. For example, while Tharros, north of the Gulf of Oristano, was under Punic control, the settlement of Neapolis, south of the Gulf of Oristano, was controlled by an indigenous population (Zucca 1987). However, the Carthaginians appeared to have interest in allowing the indige nous populations access to their trade networks. For instance, Olbia, which was founded in the fourth century BC and served as a collection point for agricu ltural products, was a point of trade with Carthage, Attica, Latium, Etruria, Marseilles, and southern Italy for the Nuragic people as well as the Carthagi nians (Antona 1991). The relationship between the Sardinians a nd the Carthaginians (and groups within the indigenous Sardinian populations) was comp lex. As previously noted, Punic material has been found in shrines, burials, and domestic sites, which is indicative of the adoption of Punic religious iconography. An example of this adoption is found at Cuccuru SArriu Cabras (Moscati 1992). Sardinian mercenaries were also recruited to fight for (e.g., at Syracuse), and ultimately against, Carthage (see Pais 1881). The returning Punicized mercenaries were able to become elites becau se of the wealth associated with the work, and they often acted as intermediaries be tween the Carthaginians and other natives (Dyson and Rowland 2007:121). Regarding health, evidence of high infant mortality of both Phoenician and Carthaginian populations was di scovered at Tharros. Five thousand infants, most less
97 than six months old, were cremated and placed in urns in an opened-air tophet, some of which dated to the eighth to seventh century BC. Three hundred of these individuals were marked by stele (Moscati 1995). It was thought originally these were the remains of sacrificed children; however, cu rrent interpretations favor a high rate of stillbirths and infant mortality (Fedele 1983; Ribichini 1989). Because of the proximity of Sardinia to the mainland, it became a desirable location for the growth of agricultural pr oducts for Rome. However, a treaty between Rome and Carthage was in place to limit the islands accessibility to Rome. Carthage did not have the power to enforce this treaty, in part because of mercenary revolts in North Africa and Sardinia. Rome seized the island in 238-237 BC, and afterward the fields of the Campidano Plain were used to feed the Sardinian populations and the expanding Roman Empire (Dyson and Rowland 2007:128). The question of when the Nuragic period ends exactly is debatable. As Rowl and (1992:175) noted, cult ural features and structures continued to be used throu ghout the Roman domination, with some being occupied into Medieval times (Webster and Webster 1998:383). 4.4 Malaria and Inherited Hemo lytic Anemias in Sardinia Malaria in Sardinia was caused by two species of the Plasmodium protozoa, vivax and falciparum which are transmitted by the disease vector A. labranchiae This resulted in a modern pattern of disease with a peak of cases and deat hs in August, September, and October (aestivoautumnal pattern), and relapses occurring year round (Brown 1984:213). A. labranchiae lives in all ecological zones in Sardinia; however, this vector shows a preference for low elevations with standing fresh water, and its populations fluctuate
98 seasonally (Aitken 1953). It is thought the climatic conditions were not favorable for A. labranchiae until about 4,000 BC, and the species probably did not take root until activities such as deforestat ion occurred (de Zulueta 1973:9-12) This disease vector has no competition on the island as it does on mainland Italy, creating a greater intensity of the disease (Sallares 2002:90). In historic times, this has led malaria to be a problem primarily in coastal and alluvial plains ar eas, with higher incidences in the Campidano and Logudoro than in the Barbagia zone (F ermi 1934; Tognotti 1997:237). Data collected from living populations by Fermi (1934) have sh own the distribution of cases of malaria does not follow this geography; however, it is not known how usef ul this is when analyzing ancient populations. Brown (1981b:366) hypothesizes the modern distribution of the disease was the result of migrati ons and gene flow (G6PD deficiency and thalassemia) during the Carthaginian and Roman dominations. The question of when and how malaria wa s introduced to Sardinia has been a troublesome one for scholars to answer. Many of the studies concerning malaria in the Mediterranean are conducted using a hist orical-ecological strategy, and often the conclusions of different researchers do not agree (e.g., Angel 1972 and Borza 1979; Brown 1984:211). Also, most resear ch has produced results that are more qualitative than quantitative regarding the effects upon human populations (Sallares et al. 2004:311). Nonetheless, malaria had severe impacts upon past populations. The most recent research on malaria in antiquity shows that it caused significant ch anges to the demography and settlement patterns of the affected populations (Sallare s et al. 2004:312). Yet, the introduction of malaria to Sardinia and its chronology have been debated and remain unresolved (Sallares 2002:90).
99 In the Mediterranean, the first mention of malaria is found in the Hippocratic texts. However, this only establishes a terminus ante quem of the fifth century for its appearance. Angel (1964:370) speculated malaria was endemic in the eastern Mediterranean long before 2000 BC, and occurred when the Neolithic package was adopted from the east. Others (Fermi 1934; Tognotti 1997:237-239) have suggested malaria spread northward from Africa to Si cily and Sardinia around 700 BC, to mainland southern Italy around 600 BC, and central Italy around 450 BC (F igure 4.9) (Sallares et al. 2004:316-317). Figure 4.9. Spread of malaria as interpreted by historic reconstruc tion (after Sallares et al. 2004; image by the SeaWiFS Project, NASA/Goddard Space Flight Center, and ORBIMAGE 2006)
100 The malarious nature of Sardinia was acknowledged by the Romans after their occupation of the island in 238 BC. Just four years later, the first report of a Roman army being destroyed by disease was made. Because of this unhealthy climate, Sardinia was used by the Roman Senate and rulers such as Nero for the banishment of enemies, prisoners, slaves, and undesirables (Salla res 2002:92-93). However, according to Brown (1984:227-229), it is possible th ese accounts were exaggera ted for political-economic reasons. For example, stating the island was unh ealthy could have serv ed as justification for the banishment of these individuals, in turn compensating for labor shortages. It should be noted lifeways of the indige nous Sardinian populations were culturally continuous during the transition from Carthaginian to Roman rule, seeing one dominator as a substitute for the other (Rowland 1977, in Brown 1984:226). Malaria in Corsica, however, appears to be more recent, occurri ng after the fall of the Roman Empire, with few references indicating its presence before the fifteenth century, and many more noting it in the sixteenth and seventeenth cent uries (Bruce-Chwatt and de Zulueta 1980:78). From a mainly historical approach, there is general agreement that malaria and thalassemia were introduced to Sardinia by people from North Africa (Proto-Sardinians), and malaria spread subsequently when the Carthaginians were trying to produce wheat on a large-scale basis on the isla nd, a process which included th e burning of forests and the creation of marshy areas favorable for the disease vector (Fermi 1934; Tognotti 1997:237-239, 2009). This resulted in cultural and subsistence modifications, such as inverse transhumance and sparse, nucleated settlement patterns in the lowland areas (Brown 1984:222-224, 1986:322-324; Webster 1996:43). Later, the introduction of slaves from Libya also resulted in more infected i ndividuals living in Sardinia and increased the
101 population density of the malarial regions furt her. Malaria became even more intensified on the island during Roman times, when more colonization (including 4,000 freedmen in 19 AD to Oristano, political ex iles, and slaves) occurred in the Campidano, Nurra, and Logudoro regions, and more land was cleare d for farming (Tognotti 1997:239). It is thought, however, that pastor alists in the mountainous ar eas were likely to remain relatively unexposed to malaria during Ro man times because the infections were occurring along the coasts and in the plains, which were used for growing grain that was exported to Rome (possibly w ith the malarial parasites), and more prevalent during the summer (Tognotti 1997:239; Sallares 2002:92-93). From historical descriptions of the disease, it is likely P. falciparum was not yet present. It has been suggested that the nuraghi (Flumene in Brotzu 1934; Sallares 2002:9192) may have been a cultural adaptation to preventing mosquito bites, not unlike the use of towers in some parts of Egypt. For exam ple, by sleeping on the upper, truncated part of the structure, people would avoid mosquito bites because the insects could not fly that high in the windy conditions of the island (Flumene in Brot zu 1934:14). Brotzu (1934:15) disagrees and argues the struct ures are better suited as de fensive towers. Brown also (1984:218) disagrees, stating the shape of th e nuraghi does not support this argument. He believes malaria (from an analysis of th e G6PD deficiency in Sardinia) was not introduced to Sardinia until about the fifth century BC, when agricultural changes, increased deforestation, and slaves were introduced (Brown 1984). Brown (1984:218) also argues the subsistence methods practiced by the Nuragic populations were primarily foraging and pastoralism with little horticulture, resulting in minimal alteration of the
102 environment that would result in a mosquito problem. As previously discussed, however, the archaeological record does not support this. For at least 2000 years, Sardinia was th e most malarious region of the western Mediterranean (Brown 1987:162).The prev alence of malaria, specifically P. falciparum and thalassemia were higher in Sardinia than in any other part of Italy, and the link between thalassemia and malaria was made because of their correlations in Sardinia (de Zulueta 1994:9). In the late 1800s, malaria caused 12% of the d eaths in Sardinia (Tognotti 1998:237). After the identification of the disease vector and before the eradication efforts star ted after World War II, the rate of malaria was reduced to 63 cases per 1,000 individuals (de Zulueta 1990:232). From 1946 to 1950, an experiment to eradicate malaria was carried out by the Ente Regionale per la Lotta Anti-Anofelica in Sardegna (ERLAAS) with help from the United Nations Relief and Rehabilitation Administration (UNRRA), the Economic Commission for Africa (ECA), and the Rock efeller Foundation (Aitken 1953; BruceChwatt and de Zulueta 1980:102-105; Tognotti 2009). This involved using DDT in an attempt to eradicate malaria-carrying mos quitoes. The species was not eradicated, however, because of a lack of knowledge about A. labranchiae. Unbeknownst to the researchers, the mosquito occupied elevations up to 1000 meters and persisted well into the interior of the island, breeding in fresh and brackish waters. Observations made during the Sardinian Project (A itken 1953:320-325) pertinent to this dissertation include that females of this species were found resting in nuraghi, that there is little evidence of springs harboring larvae (probably because of the lack of expo sure to the sun and cooler temperatures), and that mosquitoes are more intensely concentrated in rural structures
103 than in villages. Antilarval methods, such as drainage and clearing vegetation, had to be used in conjunction with DDT to rid the is land of malaria. Alt hough the disease vector was not eliminated (Tognotti 2009), this expe riment ultimately worked, ridding Sardinia of this disease, and allowing the island to be nefit in numerous ways from its malaria-free environment (Russel 1952:102; de Zuluet a 1990:231-232). In addition, the successful results of the eradication effo rts such as this throughout Ital y resulted in the widespread use of DDT in other areas of the world (de Zulueta 1994:12). It is also noteworthy that better socioeconomic conditions, such as ec onomic growth and improved standards of living, coincide with the elimination of malari a in Sardinia and other regions of Europe, even when the disease vector itself wa s not eradicated (de Zulueta 1990:235). In Sardinia, modern populations believe malaria was the result of poverty and continued economic exploitation. During et hnographic research, older individuals reported to Brown (1987:168) that they coul d not afford having malaria and would work in the field during acute episodes and drink black wine to combat the attack. Brown (1981a) describes malaria in Sardinia through folk etiological theory. According to this, the cause of the disease was bad ai r (Brown 1981a:311), and this resulted in the folk medical beliefs of Intemperie and Colpo dAria In this case, Intemperie is the idea that sudden shifts from one being hot to cold or dry to wet can cause illness. Intemperie was used to describe both the disease and its etiology, and malaria was called Intemperie Sarda (Brown 1981a:330). Colpo dAria means blast of air, and Brown (1981a) found Sardinians believe many illnesses are caused by drafts, and a person who has a fever, such as in the case of malaria, was respondi ng to a blast of cold air (i.e., the body is trying to warm itself up and restore temperature equilibrium after being chilled). Brown
104 (1981a) argued cultural adaptations to mala ria included extremely nucleated settlement patterns on higher grounds, inve rse transhumance (settlements located in the mountains with flocks traveling to the lowlands in th e winter), and the rest ricted mobility of pregnant women. The arguments for the origin of malaria in Sardinia fall into three categories, that it was introduced during prehistory (occurring with the adoption of the Neolithic package), that it was introduced through cont act with the Aegean during the Late Bronze Age and Early Iron Age, and that it was in troduced during the Cart haginian domination (535-238 BC) (Cao et al. 1989:309-312; Sallares et al. 2004:327; Sanna et al. 1997:295; Sondaar 1998). For example, Sallares et al. (2004:327) propose falc iparum malaria was introduced to mainland Italy by the Greeks around the eighth century BC, and it was probably also brought to Sardinia through Phoe nician contact, which extended into the interior of the island by the se venth century BC (Sondaar 1998). These accounts are generally taken from historic documents, and Brown (1984) advises examining the incidences of porotic hyperostosis in skelet al remains from the prehistoric to Roman periods as a more relia ble method of addressing the question of the origin of malaria in Sardin ia (Sanna et al. 1997:296). Th e earliest cases of porotic hyperostosis in Sardinia da te to the Ozieri Culture (4000-3200 BC), with the first observations of porotic hyperost osis and cribra orbitalia made in the remains of children and young men from the Bonnanaro cultu re (2200-1600 BC) (German 1995a, 1995b, 1998; Sallares 2002:92; Sa nna et al. 1997:296-297). In addition, new genetic studi es are providing more information about malaria in prehistory. For example, in Sardinia there are many adaptive responses to this disease,
105 such as G6PD (glucose-6-phosphate dehydroge nase) deficiency and thalassemia (Sanna et al. 1997:294-295). G6PD deficiency, which provides a higher resistance to P. falciparum can result in favism. Individuals with favism can de velop conditions such as anemia, jaundice, and hemoglobinuria 24 48 hours after ingesting fava beans ( Vicia faba) or certain drugs, or being exposed to certai n bacteria or viruse s. Children (usually between two and six years of age) are more susceptible than adults, and males are more susceptible than females. G6PD deficiency is a hereditary X-linked metabolic defect and occurs in a small area of the world, from the Mediterranean Basin to the Far East, with the most cases in Sardinia (Meloni an d Meloni 1997:57-62), and it is found in populations where malaria is endemic (Cappel lini and Fiorelli 2008). Several of the over 300 G6PD variants are found on the island of Sardinia: Seattle-like, Mediterranean, Sassari, Cagliari 1, Cagliari 2, Corinth, At hens-like, Menorca, Gallura, Ferrara 2, and Villasalto (Sanna et al. 1997:294-295). Because different regions have distinctive mutations, the mutations for thalassemia a nd G6PD deficiency are thought to have occurred only once and then spread by huma n migration. The first most common cause of beta thalassemia, the B+ IVS nt 110 mutation (G A), is thought to have originated in the eastern Mediterranean and preceded the or igin of malaria in Italy (Sallares et al. 2004:326). It seems most probable the CD39 mutation, the second most common cause of beta thalassemia, was likely spread from North Africa to Sardinia, Sicily, and Spain by Phoenicians in the first millennium BC, with the disease vector Anopheles labranchiae also coming from there after the Ice Age (S allares et al. 2004:326). Others believe the disease vector predates human occupation of the island by several million years (Brown 1984; Trapido 1953). This raises the questi on of whether the genetic data provide
106 information about malaria, or do they just represent migrations of groups of individuals with these balanced polymorphisms? Brown (1984:211-212) reviews th e historical reconstruc tion and summarizes the introduction of malaria to Sardinia into four testable hypotheses: 1. there was little or no malaria du ring the Bronze Age in Sardinia; 2. during the fifth century BC, malaria became a problem because of the importation of infected slaves, vast deforestation, and new agricultural methods resulting in environmental change; 3. the expansion of the latifundia system during Roman domination created changes in the ecosystem allowing for malaria to be sustained, if not increasing its presence; and 4. malaria was more prevalent in agri cultural populations in the lowlands than in the pastoral groups in the central highlands. Brown (1984:230-231) believes the best appr oach to addressing the problem of malaria in Sardinia is to exam ine mortality trends in prehistoric and historic Sardinia with paleopathological techniques, si milar to the methods of Ange l (1966, 1972). In particular, he notes Angel (1966:760) discovered pariet al bones measuring 12 mm thick on average (4 to 5 mm thicker than normal), and by us ing thickness as a measurement, one can arrive at a relative measure of the severi ty of malaria (Brown 1984:230). Comparing the prevalence of porotic hyperostos is to historical data can re sult in a greater understanding of the relationship between environment, cu lture, and economy in prehistoric Sardinia. By using the percentage of porotic hyperostos is in osteological collections over time, an indirect, relative measure of malarial intensity could be made (Brown 1984:230-231).
107 However, since Browns (1984) publication, advancements have been made in the technology used in paleopathol ogical investigations, su ch as aDNA analysis and immunological assays, allowing multiple lines of evidence to be examined when addressing this research question. 4.5 Chapter Summary Reviewing Sardinian prehistory and hist oric accounts of malaria on this island provide fundamental information for creating a re search design to test for the presence of malaria in prehistoric Sardinia. Although scholars have formulated hypotheses from historic data, Brown (1984) acknowledges it is necessary to test the hypothesis that malaria was present during the Bronze Age in Sardinia. Conditional statements addressing this problem include: 1. if there were high rates of malari a in the coastal and plain regions (especially the Campidano and Logudor o regions) during history, then there may have been a similar pa ttern of disease in prehistory; 2. if ample changes to the ecosystem (e .g., significant deforestation) occurred before large-scale wheat producti on undertaken by the Carthaginians around 500 BC, then it is possible conditions were favorable for malaria; 3. if significant contact with malarious populations occurred in prehistory, then it is possible the disease was transmitted during that time; 4. if Nuragic populations were more agri cultural rather than pastoral, then it is possible they caused the necessary modifications to the landscape, behavior, and settlement patterns for malaria to occur in a population;
108 5. if ethnographic analogies may be made from historic to prehistoric populations, then similar behaviors re garding settlement patterns and cultural adaptations may be identifi ed in the archaeological record. By examining these issues and those presented in the previous chapters with the results of a bioarchaeological analysis of skeletal data we can address the que stion of the presence of malaria in Bronze Age Sardinia.
109 Chapter 5: The Population, Materials, and Methods To test for the presence of malaria in Sardinia, a Middle Bronze Age skeletal collection from Tomb B of Serra e Sa Caudeba was examined. This group was selected because it is from the Campidano Plain (Fi gure 5.1), known historically as one of the most malarious regions on the island, in part because of its use for agriculture. The modification of this land, through actions su ch as deforestation and the planting and harvesting of crops, resulted in standing wate r, which created an ideal condition for the malaria parasites disease vector to thrive (Fermi 1934; Tognotti 1997:237). These skeletal remains were also selected because they are from a time in which it is not known if malaria was present on Sardinia, and ther e was a questionable bur ial feature suggestive of an epidemic of some sort (Soren 2003). Since malaria does not result in a specific bony response, indirect indicators of malaria resulting in bone modifi cation were used as a proxy. Th e first is the pattern of bony responses related to thalassemia (see Chap ter 3), and the second are those related to malarial or inherited hemolytic anemias (i.e ., porotic hyperostosis a nd cribra orbitalia). Regarding hemolytic anemia, it should be reite rated that porotic hyperostosis and cribra orbitalia can be representative of other condi tions (see Chapters 3 and 6); however, for the intents and purposes of th is study, the presence of he molytic anemia in cases of chronic malaria, of which porotic hyperostosis and cribra orbitalia are prime indicators cannot go without note.
110 Figure 5.1. Serra e Sa Caudeba is located on th e Campidano Plain (image by Jeff Schmaltz, MODIS Rapid Response Team, NASA/GSFC 2003). Buikstra and Cook (1980) stress the im portance of using multiple lines of evidence when analyzing diseases and stressor s in skeletal remains. In part, this is because the resulting bony response usually involves multiple nutrients or pathogens. Identifying a number of indicators to anal yze provides a better understanding of the nutritional status and stressors (including pathologies) of an individual or population (Goodman and Armelagos 1989:228). Therefore, bone samples were taken for aDNA analysis to test for Plasmodium sp. genetic material, as well as PfHRP II and hemozoin. 5.1 Tomb B from Serra e Sa Caudeba Serra e Sa Caudeba was uncovered in the early 1980s when a road was being constructed near the town of Collinas in Sa rdinia. Excavations were conducted under the
111 direction of Enrico Atzeni and Remo Forre su from 1982 to 1984. This site includes two tombs, Tomb A (Figure 5.2) and Tomb B (F igure 5.3). First, Tomb B was discovered on a knoll when a cut was made for the road, a nd then Tomb A was discovered nearby (Casu et al. 2008; personal communica tion with Mauro Perra and Alessandro Usai 2008). Both of the tombs are unusual; they have galleries like the giants tombs, but they are partially underground with the entrances bu ilt in the side of the knoll. They also lack an outer cover and an exedra. Both of these tomb s predate the large, nearby nuraghe, Genna Maria. The remains in Tomb A were discovered immediately when excavating, and they were disturbed from illicit digging. Although th e architectural style of Tomb A (Bronze Age III) is more recent than that of Tomb B (Bronze Age II), the artifacts (Appendix A) Figure 5.2. The entrance of Tomb A is built into the knoll (photo by author 2008).
112 Figure 5.3. Tomb B was discovered dur ing the construction of a road. Only half of the tomb remains intact (photo by author 2008). found in Tomb A are older than those associated with Tomb B. In both tombs the ceramics and other artifacts were few in number. Dates derived from the stylistic and compositional elements of the associated ar tifacts place the tombs in the period of 1600 BC to 1300 BC. One piece, dating to the Middle Bronze Age, c. 1400 BC, was found in a nearby home (personal communication with Maur o Perra ). In Tomb A, pottery vessels and sherds with a vi treous slip were found, indicating they might be imported from Mycenaean Greece. On the other hand, the human remains in Tomb B (Figure 5.4)
113 Figure 5.4. Tomb B during the excavation (image courtesy of Alessandro Usai) were mostly intact with little disturbance be fore the construction of the road. The artifacts from this tomb are also associated with the Middle Bronze Age. A large number of individuals were interr ed at Serra e Sa Caudeba, and although the condition of the remains make estimating sex difficult or impossible in some cases, both adults and children were present. Deceas ed individuals were interred through the small opening of the tombs and placed in an extended position close to the entrances. As more individuals were interred, and space be came scarcer, the limbs of the individuals
114 were removed and placed in the centers of th e galleries. This practice caused the remains to be highly concentrated and greatly commingled throughout the tomb. Of particular interest in Tomb B was a cist outside and to the left of the entrance of the tomb. It was made of four vertical basalt slabs arranged in a square. This feature contained the remains of subadults, including fetuses; however, th ese were not complete and were composed only of skulls and long bones. In the most adva nced level, the skulls were arranged in a circle. There were few ceramic sherds found in this tomb. The artifacts from these tombs are on display at the Civico Museo Archeologi co in Villanovaforru, which is located near the tombs (Figure 5.5), and the osteological material is stored in the laboratories of this museum. For this analysis, because of the avai lability of remains at the laboratory, only bones attributed to Tomb B were studied. Figure 5.5. The Civico Museo Archeologico in Villanovaforru (photo by author 2008)
115 5.2 State of the Previous Analyses on th e Remains from Serra e Sa Caudeba and Limitations of the Data As Larsen (2000:5) notes, one person alone does not possess the technical skills needed to complete a comprehensive study of large skeletal co llections addressing complex issues. My research with this population is only one component of the analysis of this collection. Previous and ongoing re search regarding the excavation and the skeletal collection remains unpublished, with th e exception of information (i.e., MNI, age of individuals, stature estimation, and general dental pathologies) made available online at http://www.anthroponet.it (Casu et al. 2008) The anthropologist responsible for these data was Ornella Fonzo. The basic inventory and cursory analysis were completed by two Sardinian biologists in 1992. Stature was calculated using methods developed by Manouvrier (1892). The average estimated height of males was calculated as 1.6 m, and the average height of females was estimat ed to be 1.5 m. The analysis of teeth contributed to the estimation of MNI and ag e (Atzeni et al. 2009) Although the condition of the remains is poor, prelimin ary stable isotope analysis ( 13C) of apatite is suggestive of a diet highly dependent upon me at, in particular ovicaprids; 18O ratios suggest the climate was warm and dry (Atzeni et al. 2009). Additional stable isotope research for dietary analysis is underway by Luca Lai and Robert Tykot. The formal excavation report is currently being wri tten by Alessandro Usai. Upon arrival to the museum, I met one of the individuals who had worked with the collection, Ornella Fonzo. We discussed th e previous research conducted with this collection. Unfortunately, since these data are unpublished, only certain information was provided (Tables 5.1, 5.2, and Figure 5.6). For ex ample, the teeth from Tomb B represent
116 239 individuals (81 sub adults and 158 adults ). Seventy-three of whom are under12 years of age, and 166 are older than 12. Of the indi viduals over 12 years of age, eight are under 16 (from Casu et al. 2008). Data regarding the skeletal inventories by each excavation unit, a plan map of the tombs, dental analysis, and the exact contex t of the individuals and crania analyzed are forthcoming in future publications. In this case, Fonzo advised I not duplicate work, but rather examine the skeletal elements that would answer my research questions. Therefore, this re search focuses upon the documentation of paleopathologies found in the collection, with an emphasis placed on the presence of porotic hyperostosis, cribra orbi talia, and the skeletal indica tors of thalassemia. Bone samples were also collected for aDNA, Pf HRP II, and hemozoin analysis. After I reviewed documents from the prev ious osteological res earch, I visited the curation facilities and assessed the condition of the remains and storage methods. In the curation facilities, the remains were stored in the following way. Although there were 239 individuals interred at Tomb B, only seve n relatively complete individuals (crania stored separately) were excavated. The rest of the remains were commingled. Forty-five crania, often times highly fragmented, were excavated and stored separately. No information was provided with the individuals or the crania regarding their context. Table 5.1. MNI of individuals from Tomb B Age Percentage of PopulationTotal (n) Prenatal 2.1 5 Infant 28.5 68 Adolescent 3.3 8 Adults 49.0 117 Elderly 0 0 Indeterminate17.1 41 Total sample 100.0 239
117 MNI at Serra 'e Sa Caudeba Tomb B (N = 239) 5 68 8 117 0 41 0 20 40 60 80 100 120 140Prenatal Infants Adolescents Adults Elderly IndeterminateAge CategoryFrequenc y Figure 5.6. Bar graph of the MNI from Tomb B (from the discrete ca tegories provided) Some remains were stored by excavated unit and depth. Seventy-four bags of remains had context and were stored by unit. The rest of the remains were stored by element and lacked context (e.g., box labeled vertebrae Tomb B), and included a box of bones from children and adolescents. Finally, small frag ments unidentified by the previous analysts were stored separately, without context. In some instances, however, small amounts of unidentifiable material were include d with the remains with context. In this collection, the limitations of the da ta were the result of both intrinsic and extrinsic factors. For example, the intrinsic f actors included the natu re of this secondary burial, which resulted in the commingling of skeletal elements and subsequent fragmentation of bones. The natural degrad ation of osseous ti ssue over time also impacted the quality of this collection. Previous examination of bones revealed a
118 Table5.2: Inventory of elements from Tomb B by weight Skeletal Element Weight of Fragmented Bones/Elements (kg) Crania 5.0 Vertebrae 3.0 Os Coxae, Scapulae, Clavicles 2.0 Hand Bones 1.0 Foot Bones 2.5 Long Bones 9.0 Sub Total 22.5 Undetermined 23.0 Total 45.5 substantial amount, over half of the bones in weight (Table 5.2), were not able to be identified because they were too fragmented. Extrinsic factors also placed limitations on the data collection protocol. For example, during the excavation, not all bones may have been recovered. The curation methods used with this collection also present challenges; in the case of the individuals identified during the excavation, the crania were separated from the post-cranial skeletons. Many elemen ts, as previously described, were stored without contextual information. Since contex t was not provided in all cases, the handling of the elements during documentation, prev ious analyses, or rearrangement of the curation facilities also introdu ces the possibility of error through the misplacement of bones. These previous analyses also shaped th is research in two ways. First, since some research (as previously described) was completed, efforts di d not have to be duplicated. Second, the excavation reports and findings from the previous research have not been provided as they are unpublished, and in so me cases still ongoing. Furthermore, the nature of dissertation resear ch includes its own time and monetary constraints, which result in limitations regarding data collec tion techniques and developing a comprehensive protocol to address the hypothe ses. Methods and interpretati ons were developed with the understanding of these limitations.
119 5.3 Methods of Data Collection and Analysis Used in this Research As Ubelaker (2008:1) stat es, there is no cookbook approach to commingling issues, such as those found in this population; it is necessary to form case-specific protocols addressing the partic ular issues with the populatio n and research questions. The standardization of data colle ction, however, is necessary for researchers to compare different populations. Since some research has been conducted w ith this collection, modifications were made to the standa rdized data-collection protocol in Standards for Data Collection from Human Skeletal Remains by Buikstra and Ub elaker (1994). It should also be noted that these standards were developed, in part, to address the issue of the adequate collection of data for remains subject to repatriation in the United States. Since the collection in this study is not undergoing any fo rm of repatriation, additional data can be collected at a later time to answ er other specific research questions; this, in turn, reduces the chan ce of handling the remains unnecessarily. The main purpose of this research was to test for the presence of malaria in this population; therefore, my investigation focuse d on elements that could provide the most information regarding the research question. Ot her elements were also examined for the presence of any pathological conditions, and these were al so noted. The protocol used followed Buikstra and Ubelaker (1994) a nd Kimmerle (2009) (see Appendix B). The collection process also included the creati on of a spreadsheet using Microsoft Office Excel 2003 to inventory individual skeletons and record inciden ces of pathologies observed in the individual sk eletons, crania, and remains with context. In addition, taphonomic changes were also recorded, sp ecifically focusing on unique instances and recurring examples (see Appendix B and C) Postmortem changes were identified by
120 jagged, irregular, sharp edges, while an temortem changes were distinguished by osteoblastic activity and smooth or rounde d edges (Ortner 2003:45-46). Photographs were taken using a Samsung S730 digital camera using a combination of the cameras built-in flash and natural light. A centimeter sc ale and an identification label were used in each photograph. Photographs were taken of unusual and recurring pathologies/traumas and elements useful for estimating the age or sex of an individual (Buikstra and Ubelaker 1994:10-12). The raw data and photographs of th e pathologies observed are presented in Appendices D, E, and F. The interpretations of these data are made in the following chapter. 5.3.1 Skeletal Inventories and Paleopatho logical Analysis of Individuals The analysis of the identified individual s in this population had two components. One was to take an inventory of the bones, and the other was to identify pathological conditions. During the inventory proce ss, bones with pathological conditions, modifications, unusual conditions, and postm ortem change were noted, photographed, and recorded using the standard coding (see Appendix B). All bones from an individual were identified and recorded as individual el ements, with the bones from the hands, feet, and thorax being noted as counts only. In the instance of highly fragmented remains that could not be reconstructed, such as with th e rib bones, only the num ber of fragments was recorded. Pathologies were re corded according to Buikstra and Ubelaker (1994:107-123). Only three of the seven individuals had elemen ts present that could aid in the estimation of sex or age. The element that could be us ed in the estimation of sex was the greater sciatic notch; however, one observation does not provide enough information to estimate
121 the sex of an individual, as a female could have some robusticity associated typically with males in elements used to estimate sex, or males could have gracile characteristics associated with females. Therefore, only the score was recorded and is noted with the raw data in Appendix C. The ages of the indi viduals were estimated from bone fusion, and these results are also presented with the raw data. Unfortunately, none of the skeletons was complete, and this places limitations on making differential diagnoses. Therefore, fo r these individuals, differential diagnoses were made from the elements observed. While the primary focus was to identify the skeletal pattern of bony responses associat ed with thalassemia and iron-deficiency anemia, other diagnoses are also cons idered and discussed in Chapter 6. An assessment of disease was made by examining paleopathol ogical responses on the bones. To get a general idea of the heal th of this population, the conditions were recorded using standardized coding and grouped into the categories described by Buikstra and Ubelaker (1994:112). This method of da ta collection allows for comparison with other collections and the testing of other hypotheses. The process for making differential diagnoses in this study involved the three steps described by Buik stra (1976:324). The first step was to identify the bony responses of diseases in contemporary populations. From this, a priori assumptions can be made. For this an alysis, the skeletal lesions related to thalassemia and tuberculosis (because of suggestive lesions) were included when making a differential diagnosis. The second st ep involved was identifying the diseases least likely to have caused the pathological responses and eliminating them from consideration. Finally, the results were taken into considerat ion with what is known about
122 the culture of the population to see if the charact eristics of the disease fit (i.e., does the disease reflect what we know about population concentration and diet?). 5.3.2 Paleopathological Analysis of Crania Since cranial bones and fragments were not curated with the i ndividual skeletons, they were examined separately. Only a porti on of the crania had contextual information curated with them. In all instances, the cran ial remains were highly fragmented (Figure 5.7). In some cases, portions of the cranial vault were reconstructe d previously. Because of the highly fragmented nature of the cranial bones, and the number of individuals and time constraints, only the presence of pathol ogical conditions was noted. In cases where the elements were completely present, such as in the instance of th e eye orbits, and there was no evidence of a pathologica l condition, such as cribra or bitalia, the absence of the condition was noted. However, if this portion of the bone was not present to make this observation, it was noted as indeterminate. These pathologies were also recorded Figure 5.7. Example of the condition of the cranial remains from Serra e Sa Caudeba (photo by author 2008)
123 according to Buikstra and Ubelaker (1994:107-12 3). When available, elements used to estimate sex were scored. These elements were the nuchal crest, mastoid process, supraorbital margin, glabella, and mental emin ence. Although the remains were not usually reconstructed, notes were made regarding crania l suture closure, which can be useful for estimating the age of individuals. In this study, these were limited to the sagittal, coronal, and lambdoidal sutures. The goal of this portion of the research was to identify the frequency of porotic hyperostosis and cribra orbitalia so it may be illustrated and examined using descriptive statistics. This not only provides basic information about this population, but these data are useful when comparing similar data fr om other populations (e.g., answering questions about health and subsistence practices, population fluctuatio ns, environmental changes, or contact with other populations). Since porot ic hyperostosis and cribra orbitalia are also skeletal indicators for thalassemia, the observa tions made in this portion of the research were used with the data collected from the rest of the bones in th e process of making a differential diagnosis for the disease. 5.3.3 Paleopathological Anal ysis of Other Remains In some instances, bone fragments were st ored by the excavation units and depths from which they were collected. These excav ated remains were stored in bags, with different elements being separated in the same manner as the remains without context (Figure 5.8). Each bag was reviewed, and cases of pathologies were noted.
124 Figure 5.8. Remains from Serra e Sa Caudeba Tomb B with contextual information (photo by author 2008) As discussed previously, many remains we re separated by skeletal element and had no context. For example, ribs were stored in one box, vertebrae in another, long bone diaphyses in another, and so forth. In th ese cases, since the MNI had already been calculated twice before, with agreement between the researchers, and the remains lacked exact context, the number of individuals was not recalculated. These remains were examined for pathologies, and notes on a ny observed conditions were made. The bones of the juveniles in the group were also stored in one box without context, which I believe includes the cist bu rial of interest. In rare cases, age estimations could be made by observations regarding bone fusion. These notes are included in the raw data (Appendix C). No elements used for estimating sex were observed in these remain s. The pathologies noted in these remains
125 were used with the observati ons in the individuals and th e crania to represent the pathologies present in this population. Although associations c ould not be made with the elements to reconstruct indivi dual skeletons, the combined ob servations can be useful in assessing the possible pr esence of thalassemia w ithin the population. 5.4 Collection of Bone Samples for aDNA, Pf HRP II, and Hemozoin Analysis Bone samples of suitable size and preserva tion were collected for ancient malarial DNA testing, immunology, and hemozoin analysis (Table 5.3). To have a representative sample, bone fragments were taken from each of the three natural stratigraphic levels (which were recorded during excavation as -130 centimeters, -180 centimeters, and -180 to -240 centimeters), the cist burial, rema ins without context, and an unusual feature outside of the tomb. These fragments were selected according to the recommendation Table 5.3. Bone samples collected for ancie nt DNA ,PfHRP II, and hemozoin analysis Context Bone Quantity Weights (grams) Prenatal, infants, adolescents (cist) Long bone diaphyses 4 5.7, 6.6, 6.2, 6.5 No context Long bone diaphyses 2 13.1, 13.5 Q16 (-130 cm) Long bone diaphyses 2 21.9, 11.1 Q5 (-180 cm) Long bone diaphyses 2 18.3, 6.8 Q1 (-180 cm) Long bone diaphyses 2 18.0, 12.3 Q1 (-180/-240 cm) Long bone fragments 3 3.5, 7.0, 19.4 Interments in front of the tomb Long bone diaphyses 2 14.0, 16.5 W4-X4 (first cut) Long bone diaphyses 1 16.3
126 of the Manchester Interdisciplinary Biocentre. Since comp act bone appears to be less likely to absorb contaminants, it was sugge sted diaphyses be used. Samples from the aforementioned areas with long bone diaphyses of the appropriate si ze were identified, and bone fragments were chosen randomly fr om these subsets for analysis. The total number of samples was chosen from the availability of fragments of the appropriate condition and size and the costs of analyses. The remains were placed in polyethylene bags, which was the protocol recommende d by the Manchester Interdisciplinary Biocentre. 5.5 Preparation of Bone Samples for aDNA, Pf HRP II, and Hemozoin Testing The detection of malarial aDNA, an tigens, and hemozoin was done in collaboration with and subsidization by Davi d Sullivan, Jr. at the Johns Hopkins Malaria Research Institute. To reduce the possibility of contamination, the initial preparation of the bone samples for analysis was completed by myself at the Un iversity of South Floridas Laboratory for Archaeological Science and Cell Biology, Microbiology and Molecular Biology (CMMB) Research Faciliti es, neither of which have a history of malaria research. The preparation of the samples took place in tw o phases over the span of a week. The first phase of the preparation in cluded the powdering of bone samples and incubation in 4M guanidine hydrochloride at four degrees Celsius. Ten diaphyses were chosen for analysis (Table 5.4) and photogr aphed (Appendix G). Befo re the preparation of each sample, all equipment was cleaned with a six percent sodium hypochlorite (bleach) solution and exposed to short and long wave ultraviolet radiation for two
127 Table 5.4. Bone samples from tomb B prepared for ancient DNA, PfHRP II, and hemozoin testing Sample Number Bone Age Category Context Weight of Sample (grams) 1 Humerus fragment, right Infant Bambini e adolescenti 1.57 2 Humerus fragment, left Infant Bambini e adolescenti 1.62 3 Long bone fragment, unsided Infant Bambini e adolescenti 1.31 4 Long bone fragment, unsided Infant Bambini e adolescenti 0.97 5 Ulna fragment, left Adolescent Sepoltura de fronte alla tomba B 1.24 6 Humerus fragment, left Adolescent Q1; quota 180/240 1.27 7 Humerus fragment, unsided Adolescent Q16; quota 130 0.86 8 Humerus fragment, unsided Adult Q1; quota 180 1.69 9 Humerus fragment, right Adult W4; ex primo taglio 1.12 10 Humerus fragment, right Adult Quota 177 1.50 minutes per side. Work surfaces were also cleaned with the bleach solution. Latex gloves were also worn when working with the samp les, and replaced with the handling of new specimens to reduce the chance of cross cont amination. Loose soil was brushed from the samples and sections of bone (approximately 0.5 1.5 g) were removed from each of the ten diaphyses. These samples were then e xposed to short and long wave ultraviolet radiation for two minutes per side. Afterwards, they were ground to a powder using an agate mortar and pestle. The ground bone was tr ansferred to 15 ml Fisherbrand sterile polypropylene centrifuge tubes. Approximately 10 ml of 4M guanidine hydrochloride was added to each tube. These were incubate d for 5.5 days at four degrees Celsius. After incubation, for each of the samples, half of the solution and bone powder were poured into a separate centrifuge tube. Fo r each bone sample, 2 ml of 400 mM of
128 ethylenediaminetetraacetic acid (EDTA) disodium salt was added to one of the centrifuge tubes. The samples containing EDTA were placed on a Vari-Mix (Figure 5.9) at half speed for one hour at room temperature. Af terwards, all samples were spun at 1500 xg for seven minutes. The supernatants (clear in the case of guanidine only and yellow with the addition of EDTA) were transferred to 15 ml Beckman polyallomer centrifuge tubes with Fisherbrand 5 disposable Pasteur pipettes. Distilled water was added to the supernatants to prevent the centrifuge tube s from collapsing during the subsequent 15,000 xg spin for 15 minutes in a Beckman Optima LE80 ultracentrifuge at 21 degrees Celsius at the CMMB. After centrifuging, the tubes were sealed in refrigerated packaging and sent to the Johns Hopkins Malaria Research Institute, where the supernatant and bone pellets were tested for ancient malaria DNA, Pf HRP II, and hemozoin. Figure 5.9. Processing of the samples with ED TA to maximize the chemical reaction before further testing (photo by author 2009).
129 Chapter 6: Skeletal Data and Interpretation After data collection, the results were ex amined and presented in four ways. The first component of this examination include d the overall treatment of the mortuary treatment of the population and modifications to the bones as a result of taphonomic changes. Next, the data collected from the st udy of the identified individuals at Serra e Sa Caudeba were used to make differential diagnoses. This was done in an attempt to identify indicators of inherited hemolytic anem ias, which were used as indirect evidence of malaria infections in this research. Third, the crania were examined, which included the incomplete skulls of indi viduals and elements found in association with other remains (yet not identified as being associated with a specific crania), for evidence of porotic hyperostosis and cribra orbitali a and other pathologies. Finall y, all of the data collected were examined together to identify pattern s and possible pathological conditions in the collection. 6.1 Mortuary Treatment of the Remains and Taphonomy The remains in this tomb consisted of at least three distinct phases (from stratigraphic analysis) of secondary burials. The bones in this collection were highly fragmented, which is common with secondary burials. In addition, there was discoloration (Gley 2 4/5 PB) observed thr oughout the collection (Munsell 2000). There was little evidence of carnivore, rodent, or insect activity. Aside from the highly
130 Figure 6.1. Taphonomic observations from Serra e Sa Caudeba (from upper left clockwise: insect activity, post-excavation damage, and evidence of rodent gnawing, photos by author 2008) fragmented nature of these remains, eviden ce of taphonomic change was limited to insect activity on the anterior portion of a verteb ral body, post excavation damage, and rodent activity (Figure 6.1). 6.2 The Cist Burials The remains from the cist burials were included with all of the juvenile remains interred at Serra e Sa Caudeba. In the ci st burial, a minimum of 46 juveniles were interred (Atzeni et al. 2009). Th e remains from this structure were comprised of teeth, cranial fragments, and long bones. However, dur ing the analysis it is possible that other juveniles, who may have been buried within the tomb, were included because it was not indicated that these remains were exclusively from the cist burial. Therefore, during the
131 analysis, it was not possible to determine if the juvenile re mains were interred within the tomb or in the cist burial outside of the tomb. Because the remains were very fragmented and commingled, the minimum number of individuals on whom a pathologi cal condition was observe d is presented. At least one individual displaye d evidence of porotic hyperost osis. Observation of this pathology was observed on two cranial fragment s. The condition was active at the time of death. The other pathological observation made was a fracture. This fracture was of the distal end of a humerus. Because of callus fo rmation, the fracture was not considered to be recent at the time of death. 6.3 Results of the Analysis of Individual Skeletons The identified individuals in this collecti on were not complete. Crania were stored separately in the curation facility, and it was not possible to identify the matching cranium for any of the individuals. Ther efore, the focus was placed upon identifying responses on elements present in the indi viduals in this study with the observed pathologies presented in Table 6.1. The skel etal inventory for th ese individuals is provided in Appendix C. Seven individual skeletons were identifie d during the excavation and labeled a, b, c, eI, eII, eIII, and 6. No pathological conditions were observe d on individuals a (no elements available for age or sex estimati on), eII (<18-19 years old with no elements available for sex estimation), and eIII (<14-15 years old), whose age estimations were made from radiological observations of ossification by MacKay (1961). Unfortunately, no cranial elements were associated with thes e skeletons. The incomplete condition of the
132 Table 6.1. Pathological conditions ob served on the individual skeletons Individual Bone or Element Conditions Observed per Bone or Element Age Estimation/ Scores1 Sex Estimation/ Scores2 Vertebral fragment Schmorl's node Adult Vertebral fragment Compression fracture, bone loss, osteophytes Adult Vertebral fragment Osteophytes Adult Vertebral fragment Schmorl's node Adult Fibula Myositis ossificans (unilateral) Adult Rib Periostosis 65 + 3 b Femur Myositis ossificans (unilateral) Adult Fibula Treponematosis Adult Humerus Arthritis (unilateral) Adult Vertebral fragment Osteophytes Adult Vertebral fragment Compression fracture; Osteophytes Adult Vertebral fragment Osteophytes Adult Vertebral fragment Syndesmophytes Adult Vertebral fragment Osteophytes Adult Ilium Lytic lesions Age 60 + 4 Probable male 5 c Clavicle Treponematosis Adult eI Glenoid fossa Dislocation Adult Tibiae Arthritis (bilateral) Adult Fibula Myositis ossificans (unilateral) Adult Vertebral fragment Schmorl's node; osteophytes; spicule formation Adult Vertebral fragment Schmorl's node Adult Humerus Arthritis Adult 6 Femur, head Arthritis (bilateral) Adult Probable male 5 1 In all cases where a numerical age range was not provided, estimates were made from bone ossification 2 n/a = no observable elements available for collecting sex estimation scores 3 after can and Loth (1986) 4 after Brooks and Suchey (1990) 5 Buikstra and Ubelaker (1994)
133 individuals resulted in limitations to the an alysis, including the inability to conduct a complete review of all elements used for th e estimation of age and sex, as well as making a complete examination for pathologies, which is truly needed in the differential diagnosis process to make accurate assessm ents (Kerley and Bass 1967:639; McKinley 2000:405-413). The primary purpose of analyzing these indi viduals was to determine if there was any indication of thalassemia in this popul ation; however, bony responses were also recorded in an attempt to identify other possible diseases and condi tions afflicting these individuals, and possibly the population. When possible, features that could be used to estimate age or sex were scored and recorded. However, because two of the most reliable elements used to estimate sex (the cranium a nd the os coxa) were not available (with the exception of one ilium), only the interpretations of the scores are provided to convey the possible sex of the individual. 6.4 Results of the Analysis of Cranial Bones The analysis of the 99 crania observed (Table 6.2) from this population showed evidence of porotic hyperostosis and cribra orbitalia. Additional pathological conditions of the cranial remains include bilateral hypoplas ia of the mastoid processes (n = 1, with porotic hyperostosis also observed in this in dividual), a meningeal reaction with porosity of an occipital bone (n = 1, with porotic hyperostosis and crib ra orbitalia), and endocranial lesions (n = 2, with porotic hyperostosis observed also in one of the individuals). In this study, th e earliest evidence of porotic hyperostosis was observed in the strata recorded as -180 and cribra orbi talia at -160, although not all of the remains
134 Table 6.2. The occurrence and distribution of po rotic hyperostosis and cribra orbitalia observed in the sample of crani a analyzed from this population Cranium PHCODistribution Unknown U U n/a Cranium U + Indeterminate Cranium 1 U U n/a Cranium 1* + + Indeterminate Cranium 2 + U Indeterminate Cranium 2 (l2-e2)+ + Bilateral Cranium 3 + + Indeterminate Cranium 3* + + Indeterminate Cranium 3** + U Indeterminate Cranium 4* U U n/a Cranium 5 U U n/a Cranium 6 U U n/a Cranium 6* U U n/a Cranium 7 U U n/a Cranium 7* U U n/a Cranium 8 + + Indeterminate Cranium 9 + U Indeterminate Cranium 10 U U n/a Cranium 10* U U n/a Cranium 11 U U n/a Cranium 11* + U Indeterminate Cranium 12 + U Indeterminate Cranium 12* U U n/a Cranium 13 + U Indeterminate Cranium 13* U U n/a Cranium 14 U U n/a Cranium 14* + U Indeterminate Cranium 15 + U Indeterminate Cranium 15* U + Indeterminate Cranium 16 U n/a Cranium 16* U U n/a Cranium 17 U U n/a Cranium 17* U U n/a Cranium 19 + U Indeterminate Cranium 20 + U Indeterminate Cranium 21 U U n/a Cranium 22 U U n/a Cranium 23 + U Indeterminate Cranium 24 U U n/a Cranium 25 U U n/a Cranium 26 U U n/a Cranium 27 U U n/a
135 Table 6.2. The occurrence and distribution of po rotic hyperostosis and cribra orbitalia observed in the sample of crania ana lyzed from this population (continued) Cranium PH CO Distribution Cranium 28 U U n/a Cranium 29 U U n/a Cranium 30 U U n/a Cranium 31 U U n/a Cranium 32 U U n/a Cranium 33 U U n/a Cranium 34 U U n/a Cranium 35 U U n/a Cranium 36 + U Indeterminate Cranium 37 U U n/a Cranium 38 U U n/a Cranium 40 U U n/a Cranium 41 U U n/a Cranium 42 U U n/a Cranium 43 + U Indeterminate Cranium 44 U U n/a Cranium 45 U U n/a Cranium 46 U U n/a Cranium a1 U U n/a Cranium c1 U U n/a Cranium C2 + + Indeterminate Cranium c-e U U n/a Cranium d + U Indeterminate Cranium d1 + U Indeterminate Cranium g U U n/a Cranium g1 U U n/a Cranium ie1 + + Bilateral Cranium l1 U U n/a Cranium m1 U U n/a Cranium M1 + U Indeterminate Cranium O1 U U n/a Cranium p1 U U n/a Cranium q + U Indeterminate Cranium r1 U U n/a Cranium s U U n/a Cranium t U U n/a Cranium t* U n/a Cranium t1 U U n/a Cranium t2 + U Indeterminate Cranium v U U n/a Cranium z + U Indeterminate Unnumbered A + U Indeterminate
136 Table 6.2. The occurrence and distribution of po rotic hyperostosis and cribra orbitalia observed in the sample of crania ana lyzed from this population (continued) Cranium PH CO Distribution Unnumbered B U U n/a Unnumbered C U U n/a Unnumbered D + + Indeterminate Unnumbered E + + Bilateral Unnumbered F U + Bilateral Unnumbered G U + Indeterminate Unnumbered H U U n/a Unnumbered I + + Bilateral Unnumbered J U U n/a Unnumbered K + U Indeterminate Unnumbered L U U n/a Q11 (-160) Cranium 13 + + Bilateral Q11 (-160) Cranium 15 + U Indeterminate Q6 (-180) Cranium 3 + U Indeterminate Q6 (-180) Cranium 5 U U n/a CO refers to cribra orbitalia PH refers to porotic hyperostosis U indicates the element was not present for observation + indicates the condition was present indicates the condition was absent duplicate numbering for cranium ** triplicate numbering for cranium have contextual information, so it could be possible these conditions could be found in remains from the deepest levels. In the fragme nted crania studied, 38 of the 99 skulls had evidence of porotic hyperostosis (n = 23), cr ibra orbitalia (n = 4), or both (n = 11). Therefore, at minimum, 23.2% of this sample had porotic hyperostos is, 4.0% had cribra orbitalia, and 11.1% demonstrat ed both conditions, with a tota l of 38.4% of this sample showing evidence of these conditions (Figures 6.2 and 6.3). Since the crania were incomplete, it is likely these percentages are higher. In all cases of cribra orbitalia when it
137 Porotic Hyperostosis and Cribra Orbitalia Observed in Crania from Serra 'e Sa Caudeba Tomb B (n = 99) 4 23 11 61 0 10 20 30 40 50 60 70 80Cribra Orbitalia Porotic Hyperostosis Both Conditions No Indication of CO/PHConditionObservation s Figure 6.2. Crania observed with and without po rotic hyperostosis and cribra orbitalia Frequenc y of Porotic H y perostosis and Cribra Orbitalia Observations by Bone (n = 80) 42 21 7 9 1 0 10 20 30 40 50 Cranial Fragment (Unidentified) Frontal BoneOccipital BoneParietal BoneTemporal Bone BoneFrequenc y Figure 6.3. Distribution of case s of porotic hyperostosis and cribra orbitalia by bone (the number of frontal bone observati ons is the total for both conditions)
138 was possible to observe both orbits of the frontal bone, the condition presented itself bilaterally. 6.5 Overall Osteology Results Including Post cranial Bones, Individuals, and Crania Since there were few individual skel etons available on which to perform differential diagnoses, and those remains we re not complete, the entire collection was observed for conditions that mi ght provide clues about the he alth of this population. Bony responses were documented and used to addr ess the hypothesis that malaria was present during the Bronze Age in Sardinia. The observa tions of pathologies are categorized and presented in Figure 6.4. A more detailed description of these responses is included in the discussion in the following chapter within the context of tes ting the hypothesis. Summary of Pathological Conditions at Serra 'e Sa Caudeba Tomb B (n = 180) 13 40 4 22 6 80 15 0 10 20 30 40 50 60 70 80 90Abnormal Bone Formation Abnormal Bone Loss Abnormality of Shape Arthritis Fractures and Dislocations Porotic Hyperostosis/Cribra Orbitalia Vertebral PathologyCategoryFrequenc y Figure 6.4. The frequency of the pathologica l categories observed at Serra e Sa Caudeba Tomb B (after Buikstra and Ubelaker 1994:112)
139 6.6 Results of Malarial aDNA, Pf HRP II, and Hemozoin Analysis from Serra e Sa Caudeba Tomb B Polymerase chain reaction (PCR) is used to study ancient DNA because it is a sensitive method requiring very small amount s of DNA; however, the sequence of the DNA to be studied must be known. In the case of the aDNA analysis with the remains from Serra e Sa Caudeba Tomb B, the initial results produced positive results (i.e., samples four, seven, and nine tested positive for falciparum malaria, and samples nine and ten tested positive for vivax malaria). Additional attempts to replicate these results failed. Malaria infections also result in the presence of Pf HRP II, as previously discussed in Chapter 3. Rapid tests are sometimes used to identify this protein in remains from archaeological contexts. In the case of this study, the samples were tested using western blots. Protein was observed, but the tests were negative for Pf HRP II (personal communication with David Sullivan, Jr. 2009). Tests for hemozoin were conducted using LDMS in the Cole Laboratory at Johns Hopkins University. There were no results s howing clear evidence of hemozoin. Sample five (with EDTA) produced non-conclusive peaks, and the rest demonstrated lowamplitude background noise (personal communication with John Pisciotta and David Sullivan, Jr. 2009). 6.7 Conclusions The juvenile remains in this collection, including those from the cist burial, showed evidence of three cases of bony responses. Two cran ial fragments with evidence
140 of porotic hyperostosis were observed, and one humerus showed evidence of a healing fracture. The identified individuals in this population, which were incomplete skeletons, showed evidence of arthritis, with repeated observations being made on vertebral elements. Evidence of trauma was obser ved on a scapula (a possible shoulder dislocation).The gross examination of the crania showed high rates of evidence of porotic hyperostosis alone (23.2%), wh ile 4.0% of the individuals had evidence of cribra orbitalia. Both conditions were observed on 11.1% of the crania. A total of 38.4% of the crania were observed to have evidence of poro tic hyperostosis, cribra orbitalia, or both conditions. Overall, the most common categ ories of pathologies observed included porotic hyperostosis and cribra orbitalia, abnormal bone loss, and arthritis. Evidence of six fractures and dislocations were observed. The aDNA analysis produced four cases of positive results that could not be replicated. Protein analysis did not demonstrate any evidence of Pf HRP II. In addition, hemozoin was not detected.
141 Chapter 7: Discussion The review of the literature pertaining to malaria, paleopathology, and Sardinian prehistory provides a foundation for identifyi ng conditions to formulate an approach to test the hypothesis that malaria was presen t in Sardinia during the Bronze Age. These conditions can be classified into three categ ories of evidence: archaeological/ecological, genetic, and skeletal. To begin, a discussi on of the theoretical underpinnings of the biocultural approach is presented. Next, th e ecological and archaeological data are presented and compared to the previous hypotheses and statements made by other scholars regarding malaria in Sardinia, such as the suitability of the environment for the disease vector, the lifew ays of the prehistoric people, c ontact with other populations, and possible prehistoric cultural adaptations to a malarious environment. Ethnographic evidence, which was presented in Chapter 4, is compared with archaeological evidence to identify analogous behaviors that could be adaptations to a malarious environment. Finally, the inherent nature of osteological data and the methods used in this study are examined. 7.1 The Biocultural Approach in Anthropological Research When studying disease vectors, it is nece ssary to understand changes in the host or pathogen, as well as the environment. In human populations, this environment includes culture (e.g., social systems, ideology, architecture, technology) (Armelagos and Dewey
142 1970:271). Inhorn and Brown (1990:89) observe diseases have been a prime mover in cultural transformation. In some cases, cultural aspects may promote infection, while others prevent it. To understand these relationships, biocultural approaches are often used. Biocultural approaches integrate biologi cal, cultural, and ecological information to understand humans (McElroy 1990:244). In the 1960s, these approaches were used when research topics started to include tes ting hypotheses or addres sing problems, rather than focusing on descriptive methods. This new perspective focused frequently on human adaptability in various environments. Methods were developed through studies addressing response mechanisms to environm ental stressors, which were sponsored by the Human Adaptability Projec t of the International Biol ogical Program of the 1960s (McElroy 1990:246). Some examples of biocultural approaches relating to malaria include Ladermans (1975) use of historical documents to recons truct the spread of malaria through the Near East and the Medite rranean, and Browns (1981) use of various lines of data to understand biocultural adap tations to malaria observed in historic populations (as discussed in Chapter 4). Most infectious disease research has been ecological, that is, it focuses on the host and agent within the ecosystem. Medi cal geographer May (1958) wrote The Ecology of Human Disease in which he presented a model of th e interactive role of environments, both physical and sociocultural, when studyi ng infectious diseases (Inhorn and Brown 1990:95-96). Many scholars followed this approach. For instance, Audy (1971, 1974) expanded upon this with the intr oduction of insults (i.e., infecti ous, chemical, or social) to the disease process, and Dunn (1976a, 1976b, 1983, 1984) added to this by combining
143 complex interactions, such as those between host, biology, behavior, and environment, into what he called causal assemblages. From these, sociocultural approaches emerged. Turshen (1984) presented the political ecology of disease as a way of understanding poor health, which uses Marxist theory, and includes examining political, economic, and social issues when studying disease. These anthropological studies of infectious di sease usually include both a macrosociological perspective and a microsociological perspective (culturally prescribed behaviors of an individual that can result in risk or preventio n of an infectious disease) (Inhorn and Brown 1990:98-99). An other pioneer in this field, Alland (1970), used evolutionary theory to study how cultural behaviors aff ect human hygiene and health (Inhorn and Brown 1990:99). Later, D unn (1976, 1979, 1983) generated a model identifying health behaviors in two ways, those promoting health, and those demoting health (as not all human behavi or is adaptive in the evoluti onary sense). These behaviors can be classified further as deliberate or non-deliberate and health-promoting (or maintaining) or health-demoting (Inhorn and Brown 1990:99). Therefore, these healthrelated behaviors fall into four categories: (a) deliberate health-re lated behavior that promotes or maintains health; (b) deliberate behavior that contributes to ill health or mortality; (c) behavior not pe rceived to be health related that enhances or maintains health; and (d) behavior not perc eived to be health related that contributes to ill health or mortality. Dunn (1976, 1979, 1983) divides thes e categories further by including the perspectives of insiders (the population at risk) as opposed to outsiders (members of the health-care community). For example, Wood (1979) observes many studies of the
144 interaction of cultural practi ces and malaria in endemic areas show oftentimes cultural practices limiting the transmission of this disease are not deliberate. More recently, in the volume edited by Goodman and Leatherman (2001), the need for a joining of sociocu ltural and biocultural theories is addressed. In particular, they focus on the political economy of healt h. However, Dressler (2005) believes the development of research methods and an el aboration of theory are needed. While a definition of political economy is presented, th ere is a need to identify what is cultural and what is not in this process. It then follows that it is important to define terms such as adaptation and culture. For the intents and purposes of this discussion, adaptation will refer to the selection of genetic (such as balanced polymorphisms) or cultural traits (Alland 1966) increasing a populations chances of survival through successive generations in a given environment (McElroy 1990:249; McElroy and Townsend 1989:73, 76). In particular, the term adaptive st rategies is used to describe cultural characteristics that enhance survival (McE lroy 1990:249). Culture will be defined as the human, extra-somatic means of ad aptation; in this case, that which is represented in the archaeological record (Binford and Binford 1966; Binford 1973). 7.2 Ecological and Archaeological Evidence of Malaria in Nuragic Populations 7.2.1 Is there evidence of cultura l adaptations to malaria in Bronze Age Sardinia? As discussed in Chapter 4, it was suggested by Flumene (in Brotzu 1934:14) and Sallares (2002) that the nuraghi provided protection from a malarious environment. However, Brown (1984:218) disagreed by arguing that the sh ape of the nuraghi does not support the argument that these structures were truncated to provide elevated sleeping
145 levels to avoid mosquitoes. Interesting evidence was produced during the Sardinian Project (Aitken 1953:320-325). This research, which included an analysis of the disease vector, found female mosquitoes resting in th e nuraghi. This observation is also contrary to the idea the nuraghi provided protection from mosquitoes. However, it is also possible there may have been additional cultural modifications made to these structures while they were occupied that could have prevente d mosquitoes from entering the towers. Archaeological interpretations (Guilaine 1992; de Lanfranchi 1992; Lilliu 1967) point to the towers being adopted because of cultural influences from the north (mainland Europe and Corsica), where it is not likely there was malaria. At most, they would have provided some unintentional benefit. More interestingly, yet not discussed previously by scholars, is the possibility that Nuragic water management practices, such as sacred wells and other enclosed sources that restricted exposure to sunlight, were a biocultural adapta tion to a malarious environment. The research from the Sardinia n Project found little ev idence of mosquito larvae in springs and even less in wells (Aitken 1953:320-325). The lack of larvae in wells and springs was attributed to an absence or minimal amount of sunlight and thus cooler conditions. Other water-management tec hniques, such as the use of systems of channels, fountains, and the storage of coll ected water, within the Nuragic complexes may also have provided health benefits in a malarious environment. If malaria was indeed present during the Nuragic period, additional research focusing on the water-management practices of this time could provide information about the possible biocultural benefits of these behaviors.
146 7.2.2 Did sufficient changes to the ecosystem o ccur to support the disease vector before or during the Bronze Age? For malaria to have been present in Sa rdinia, the environmental conditions must have been favorable for the disease vector, A. labranchiae to have survived. Earlier interpretations argued that the disease vect or was not supportable on this island until about 4000 BC (de Zulueta 1973:9-12). However, with new knowledge about the disease vector and subsequent research, more recent data point to the vector predating human occupation of the island by several million y ears (Brown 1994) or, at least, coinciding with the earliest human occupations (Salla res et al. 2004:326). Furthermore, evidence has shown A. labranchiae inhabited elevations up to 1000 f eet, which would include most of the island. The amount of ecological data available for Sardinia is scant. There is an indication of substantial defore station at the end of the Ne olithic with a warming trend around 2500 BC (Vigne 1990:377; Whittle 1996) During the Middle Bronze Age (16001300 BC), people increasingly used metal increa singly. Lai (2008) posits it is likely this technology contributed to defore station, because trees would need to be removed to gain access to the ore, and wood would be used for fire in the smelting process. It is also likely trees were used to aid in the movement of stone during the construction of nuraghi and giants tombs, as fuel when making pottery and cooking, as well as the manufacturing of seafaring vessels. However, more research is ne eded to verify if, and the degree to which, this occurred.
147 7.2.3 Did subsistence patterns of Nuragic popu lations result in land modification conducive to malaria? It is commonly thought Nuragic populati ons primarily practiced pastoralism. However, as more archaeological evidence is discovered and isotopic studies (Lai 2008) are conducted, it is likely that this is a misconception. While a subsistence pattern relying primarily on pastoral activities would lim it deforestation and reduce the standing water associated with agricultural activities, it is not likely that this was occurring at this time. As previously noted, Lazrus (1999) points out that there is no archaeological evidence indicating an increase in pastoral activiti es (e.g., textile produc tion, dairy products, or structures for animals), and furthermore, the settlement patterns are more indicative of a culture utilizing a diversif ied, mixed subsistence strategy, one relying both upon agriculture and pastoralism. Therefore, subsistence methods used during the Bronze Age also had an impact on the modification of the landscape. Evidence supporting the continuing use of agricultural methods woul d mean there was deforestation, soil erosion, and areas of standing water conducive to mos quito breeding. In addition, isotopic studies by Lai (2008:283-284) do not supp ort the hypothesis for a foragi ng/pastoralism model in reference to the Early Bronze Age. Future isot opic research for samples from the Middle and Late Bronze Ages can provide more detail about subsistence practices. 7.2.4 Is there evidence of contact with other populations that may have been malarious before or during the Bronze Age? Similar material culture is often th e result of contact between populations (Sherratt 1972:525; Struever and Houart 1972; Hodder 1978, 1979). The quality and
148 degree of this contact is thought to infl uence artifact similarity (Cohen 1977:82, Hodder 1979:446). For example, artifacts with unique st yles may be used for self-identification purposes when tensions exis t between groups (Hodder 1979:450). The archaeological evidence from Sardinia shows the island was part of an exchange network with other is lands and mainland Europe star ting in the Early Neolithic. For example, obsidian attributed to Monte Arci has been found in Corsica, mainland Italy, southern France, and as far as Spai n (Figure 7.1). In addition, impressed wares common in the Iberian Peninsula, France, It aly, North Africa, and Sardinia may have been obtained through the exchange of goods or technology, skills, or people, as noted by Dyson and Rowland (2007:28-31). Trade continue d in the Middle Neolithic. The pottery Figure 7.1. Distribution of obsidian from Monte Arci during the Neolithic (reprinted with permission from Tykot 2004)
149 from this time also shared stylistic attri butes with pieces found on mainland Italy, Sicily, and the Balkans (Dyson and Rowland 2007:34). Furthermore, the Late Neolithic tombs on Sardinia share similarities to thos e found on Malta and Sicily (Contu 1964). The presence of Bell Beakers in Chalcolithic tomb s have been interpreted as evidence of an intrusive group or circulation of exotic prestige items (Balmuth 1992:676). Goods and technologies such as olive oil, Mycenaean pottery, Cypriot copper, and casting techniques show evidence of Bronze Age contact with the Aegean and Near East, both of which may have been malarious during this time (Angel 1964; Knapp and Cherry 1994: 146-151; Lo Schiavo 1989a). 7.2.5 Can ethnographic analogies be made to Nuragic populations? Brown (1981a) describes several cultural adaptations to malaria in populations living in the 20th century. These included wine consumption to combat attacks; the reduction of drafts, shifts in temperatur e, and wet and dry conditions; extremely nucleated settlement patterns; inverse trans humance; and the restricted mobility of pregnant women. However, of these, th e only one that can be supported with archaeological evidence currently is the occu rrence of nucleated settlement patterns, which is in the form of Nuragic villages. Although wine was consumed during prehistory, it cannot be inferred it was used to combat malarial fevers. Currently, there is no archaeological evidence for the restricted m obility of pregnant wo men (e.g., in artwork), or of inverse transhumance. The nuraghi could have provided protection from the elements (e.g., drafts, rain, and temperature changes); however, this is common of most shelters. Therefore, it is not likely analogies can be made at this time, beyond noting the
150 nucleated settlement patterns. However, th is does not mean future archaeological research cannot be developed to create and address hypotheses related to these observations. 7.3 Discussion of Osteological Research When conducting paleopathological analyses one question that is crucial to the interpretation is how similar is the disease pr ocess of today to that in the past? To begin, diseases present today may not have been in the past, and vice versa (Manchester 1987:163-170). For archaeologists to assess disease in past populations, they must look for evidence of it in literature, art, and huma n remains. When the presence of a disease is identified, we can then begin to address the factors that encouraged or prevented the development of a disease, as well as social aspects of the disease and its relationship to the environment. As discussed previously, osteons can onl y respond in a limited number of ways: they can be formed, reabsorbed, or some combination of both (Buikstra and Cook 1980:439; Cohen 1989:106; Goodman and Armela gos 1989). Therefore, the response of bone to stress or disease is also restricted and predictable; paleopathological conditions involve formative lesions, resorptive or lytic lesions, or a combination of these types of lesions, and these responses are typically f ound in specific locales on the skeleton or a pattern that develops within a population (Cohen 1989:106; Ortner and Putschar 2003). This aspect of bone allows us to understand a nd identify various stressors such as disease, trauma, and improper nutrition. However, many diseases look alike, and others leave no skeletal indicators. Skeletal samples provide only a lim ited representation of the
151 individuals health. The absence of associ ated soft tissues, and the principle of equifinality in this case, many conditions can create similar skeletal lesions present challenges (Mitchell 2003:175). In some instance s, diseases that w ould cause lesions on the skeleton may cause death in certain indi viduals before the bones can respond, and this can create a bias for chronic diseases, whic h have a longer time to affect the bones and generate a bony response (M itchell 2003:175). This can also result in an under representation of disease or trauma (Cohen 1989:106). Since analogies are made from observations of bone in living populations, the result may be a broad or general diagnosis that is similar to a modern disease, which is not always the best interpretation (Cohen 1989:107). Unfortunately, most diseases, including epidemics, do not result in skeletal responses that may be iden tified specifically (Cohen 1989:109). For more detailed information, Ortner and Putschar (2003) a nd Buikstra and Ubelaker (1994) provide good references for paleopathologies, trauma, cu ltural bone modification, taphonomic changes, and data collection standards and technique s. Ortner (2003:109-118) presents five theoretical issues in paleopathology: 1. when a disease is observable in skelet al tissue, it most likely represents a chronic, long-term condition th at afflicted an individual; 2. in general, paleopathological knowledge is developed from historical data, archaeological remains, and not experiments; 3. as biomedical research and knowledge about the human past advances, there is a constant modification of how skeletal diseases are classified; 4. inferences about the health status of a population are interpreted through observations made when studying skeletal remains, with the objectives of
152 paleopathological research being to understand adaptive responses to new or evolving pathogens and the impact of disease; 5. there are limitations to paleopathologi cal research. Skeletal manifestation of a disease is one of the ranges of possibilities of a disease state, falling somewhere between a disease resulting in a rapid death and one in which the individual recovers with healed lesions leaving no evidence of the disease. Also, in only a few cases will an individual with an infectious disease show any skeletal response. Furthermore, gender differences play a role in the skeletons reaction to disease (e.g., maternal nutrition and mortality, gender differences in nutrition, morbidity, and immune responses). In general, the problems associated with using skeletal remains to interpret the past are similar to those of all archaeological evidence. Th ese challenges include sparse records, poor preservation, and incomple te recovery of samples (Cohen 1989:105). Waldron (1994:11-17) and Lewis (2007: 24) note that there are factors influencing the recovering of remains, and in fact, they ar e almost never random. These factors, both extrinsic and intrinsic, creat e biases in the interpretati ons of paleopathological and paleoepidemiological studies when trying to apply the results to the whole population. Extrinsic factors include the proportion of the dead buried at the si te, the proportion lost by disturbance or factors infl uencing preservation (e.g. gr oundwater chemistry, clothing, soil type temperature, oxygen levels, depth of burial, and flora and fauna), the proportion discovered, and the proportion excavated. For example, skeletal remains may only be a selected sample of the population they repres ent. Formal burials, or for that matter the
153 ones recovered in a specific area, may represen t a selected group or status of individuals. A population may not include indi viduals of a certain age, status, or lineage in their formal burials (Cohen 1989:106). Saunders (1992:2) observes that infants (under one year of age) are not usually bur ied in cemeteries, but rather in other contexts. Overall, the funerary treatment of non-adults varies. In some cases, there is evidence of ritual and consideration of their placement, and in othe r cases remains are found in middens (Lewis 2007:37). The intrinsic factors ar e the effects bone size, shape, density, porosity, and the age of the bone have upon preservation; furt hermore, the population examined is a dead population, which may not exhibit conditions that are representative of their cohorts. Another challenge one faces when working w ith skeletal collections is related to when and where the remains were excavate d. Archaeological interest and focus has changed throughout time and varies by geographi c region. This also presents challenges when studying skeletal remains. For example, earlier excavators may not have been as interested in skeletal material, and they may have placed less emphasis on recording the context of human remains, their analysis and curation (Cohen 1989:106). Understanding the lifeways of the culture being studied is in tegral for the interpreting the disease process within a population, and a knowledge of the mortuary practices is important so biases in the data may be recognized by the paleopathol ogist, as burials may ha ve been segregated by the health or age of the indivi duals (Buikstra and Cook 1980:443-444). Because of perceived problems concerning preservation and recovery, the use of subadults is often overlooked in research; however, they are quite valuable. It is commonly thought the bones of children do not preserve as well as the bones of adults; however, given the right circumstances, juvenile remains can display excellent
154 preservation (Lewis 2007:20). When soils are highly acidic or alka line, juvenile bones may not preserve as well; soil pH has been found to be significantly correlated to bone preservation, especially in subadult bone s (Gordon and Buikstra 1981:589). This is because subadult bones are typica lly less dense, more fragile, and composed of a higher organic content and lower mineral content than adult bones, which makes them less likely to preserve as well as bones from adults (Currey and Butler 1975; Specker et al. 1987). Sundick (1978) on the other hand, argues the i ssue is not necessarily the preservation of subadult bones, but rather a refl ection of the skills of the excavators during data recovery (Saunders 1992:2). In addition juvenile remains become disa rticulated more easily than adult remains, and cranial bones are more fragile, which could result in more postmortem damage (Haglund 1997). While it is more difficult, if not impossible to determine the sex of these individuals (w ith the exception of identification through DNA analysis in some cases, and possibly through so me promising advances with the analysis of deciduous teeth dimensions and mandibul ar features [Lewis 2000:52, 2007:13,58]), it is much easier to determine their age than it is with adults. This is especially useful when comparing growth rates, nutritional factors, variation in stressors, and paleodemography (Baker et al. 2005:3-5; Lewis 2000:39). Their inclusion is necessary in establishing a representative sample for addressing paleode mographic issues such as mortality and survivorship, life expectancy at birth, and popul ation size and structure. In particular, for this research, the analysis of subadults is valuable when studying diseases such as thalassemia (which presents in a specific pattern of bony res ponses in the skeleton), when many homozygous individuals may not su rvive past childho od without medical
155 intervention (Baker et al. 2005:4) It should be recognized that children in this context, however, represent the non-su rvivors (Lewis 2000:40). While it is desirable to have juveniles represented in a skeletal population when assessing health, especially in the case of diseases resulting in death early in life, there are challenges associated with analyzing juvenile remains. For example, when examining pathologies, it is important to understand th e distribution of the manifestation of the disease throughout the skeleton is different in children than it is in adults (Lewis 2007:162). This is because of a greater di stribution of hemopoietic marrow throughout the skeleton, so infections such as tubercul osis or osteomyelitis may be found in areas of the skeleton where it would not be occur in adults. In juvenile bones, the periosteum is more loosely attached, so infections may a ppear more wide-spread. Also, since the bones are more porous, abscesses may be harder to identify. Buikstra and Cook (1980) stress the im portance of using multiple lines of evidence when analyzing diseases (stressors) in skeletal remains. In part, this is because skeletal responses usually involve multiple nut rients or pathogens, and it is difficult to diagnose specific diseases from dry bone or radiographs alone (Miller et al. 1996). Therefore, creating a number of indicators that can be analyzed provides a better understanding of the nutritional status and stressors of an individual or population (Goodman and Armelagos 1989:228). For exampl e, stable isotope analysis (personal communication with Luca Lai 2009) and mala rial aDNA used in conjunction with the gross osteological examination can provide a more complete understanding of the health and lifeways of these people.
156 7.4 Discussion of Skeletal Eviden ce from Serra e Sa Caudeba Tomb B 7.4.1 Mortuary Treatment of the Remains and Taphonomy A dominant processual approach used toda y to interpret mortuary remains is the Saxe-Binford approach (Rakita and Buikstra 2005b:5), which finds a relationship between mortuary treatment and social status (the more energy expended on the individual, the higher the soci al status) and states mortua ry treatment and rituals are correlated to the subsistence strategy (which was used as a proxy measure of social complexity) (Binford 1971). Saxes Hypothe sis Eight links areas of formal burial or interment to territoriality (Saxe 1970). The purpos es of tombs have al so been interpreted as being attention-focusing devices and a place to perform ceremonial rituals while reinforcing leadership patterns (Flemi ng 1973). Tombs, burial mounds, and other funerary monuments might also be symbolic or represent territorial ity between competing groups or lineages within th e same location (Renfrew 1976). One aspect of this collection that made it desirable for this study was the cist burial outside of the tomb containing only juve nile remains. It is an unusual feature in Sardinia, and the treatment of the remains dur ing burial could be a fo rm of veneration or violation related to an epidemic. Violation a nd veneration are cultural constructs that may change within a culture and over time. Some cases of violation may result in the creation of symbols from the deceaseds remains, and these may be found in later burials as venerated objects (Bloch 1992; Duncan 2005:21 0). Veneration is an act that helps the soul rest or honors the memory of an individual. This action is usually reserved for those joining ancestors or high-status individuals. Violation, on the other ha nd, is an action that denies a resting place for the deceased or dest roys their soul; this usually occurs when the
157 behavior of the individual was socially un acceptable or cause of death was undesirable, such as in the case of disease or suicide. Cross culturally, veneration and violation can look similar in the archaeological record, because they both use destruction in the treatment of the rema ins (Duncan 2005:207). Weiss-Krejci (2005) and Ra kita and Buikstra (2005a :94-95) discuss secondary burials and possible explanations to this pr ocess, such as political, economic, or biological, and concludes sometimes mortuary treatments will not allow us to make a conclusion about the status. In particular, she (Weiss-Krejci 2005) addresses various treatments of corpses when death occurs aw ay from the ultimate burial location resulting in a delay of interment, or when numerous people die in a short period of time. Regarding the color changes observed on the bone, a similar color was observed by Piga et al. (2008), in the Early Bronze Age tomb of Sa Figu Tomb IX, and the remains were interpreted as being possibly cremated or otherwise burnt by a subsequent fire. The condition and color of cremated bone can ra nge from slightly burnt (brown/black) to oxidized (buff/white). Since cremation and secondary burial often result in fragmented remains with elements missing, it is often difficult to estimate the sex or age of individuals. In addition, this also places limits on what can be assessed pathologically, since complete skeletons are often necessary for reliable diagnoses (McKinley 2000:405413). Cremations in Sardinia in the Bronze Ag e are very rare, with one possible example occurring in the Early Bronze Age tomb of Sa Figu Tomb IX, although researchers are uncertain if the condition of the bones was the result of intentional cremation or natural fires after the tomb collapsed (Piga et al. 2008). Other taphonomic changes were limited
158 to insect and rodent activity, which could have occurred before excavation or after curation. 7.4.2 Discussion of the Cist Burial The disarticulation and arra ngement of bones and skeletal elements in the cist burial is indicative of a form of violation or veneration. Ho wever, it is important to consider that in some cultures infanticide, which may be observable in the archaeological record in terms of veneration or violation, may occur under the premise that infants were not yet given the same status or value as older individuals (Kam p 2001; Scott 1992). The mortuary treatment of these remains, as noted before, is consistent with the occurrence of an epidemic. However, the question remains, if there was an epidemic, could it have been malaria? The remains most indicative of a malarial infection in this sample would be the cranial elements. In the case of thalassemia or persistent malarial anemia, which could occur with P. vivax infections, the most common condition observable in these fragmented remains would be porotic hyperostosis. The data collected during this research suggested that one individual, at mini mum, displayed evidence of this condition. Without microscopic or radiographic analys is, it is not possible to determine if these lesions are related to infection or to suggest what disease was afflicting the individual. However, pertaining to malaria and inherited hemoly tic anemias, this could be interpreted in two ways. First, it could suggest that there is no indication of recurring P. vivax malaria episodes; however, it does not rule out death during the early stages of a P. vivax infection or P. falciparum malaria, which could result in fatality before
159 macroscopic changes to the bone tissue coul d occur. Second, it could suggest that an inherited hemolytic anemia was not a condition that was affecting this population; however, it is also possible that the juven iles in this population, who represent the nonsurvivors, died before skeletal evidence of this condition could have developed. If this was the case, it is likely that only those surviving past early childhood would show evidence of porotic hyperostosis and cribra orbitalia. Regarding these specific bony responses in older individuals, the results of this research are consistent with that argument. The other condition observed was a fracture. This fracture was of the distal end of a humerus (supracondylar fracture), which is not an uncommon injury in children, and it usually results from falling on an outstretched hand (Marrow et al. 2007). Therefore, it is likely that this fracture is accidental and not indicative of violence. 7.4.3 Differential Diagnosis from Individual Skeletons As presented previously, thalassemia affect s the skeleton in a distinctive way that can be identified through a process of diffe rential diagnosis. Since the remains in this collection were highly fragmented and commingl ed, not all of these indicators could be observed. For example, because there were no completely intact crania, observations could not be made regarding characteristics such as malocclusion or prognathism, which are both found in thalassemic patients. The data collected and presented (Appendi ces C and D) demonstrate there is no observable evidence of thalassemia in these i ndividuals. However, it is also proper to note many individuals who have thalassemia majo r would not be likely to live as long as
160 these seven individuals did, and in the even t they had thalassemia intermediate, the missing crania would more than likely be needed for measurements and observations. Nonetheless, the observations made provide clues to Nuragic life. The pathologies observed in these individuals mainly fall in to the categories of arthritis, vertebral pathologies, and bone loss. There are two excep tions to this, perios titis observed on a rib fragment from individual b and multifocal ly tic lesions on the ilium from individual c. The pathology on the rib could be indicative of a lung infection, and the lesions on the ilium could be the result of numerous conditions, such as cancer, tuberculosis, or brucellosis (Aufderheide a nd Rodrguez-Martn 2005; Bu ikstra and Ubelaker 1994; Ortner 2003) which was found in later Roman sites and attributed to the consumption of untreated sheep and goat milk (Capasso 1999). However, without other evidence in the skeleton, it is impossible to make a diagnosis in these cases. For example, arthritis is noteworthy; however, the nature of these ar thritic conditions (e.g., rheumatoid arthritis, osteoarthritis, tuberculosis arthritis) cannot be interpreted beyond a general classification because not all elements are present. 7.4.4 Results and Interpretation of the Analysis of Cranial Bones Most porotic hyperostosis and cribra orbi talia occurs in children. In cases with unhealed lesions, the individual is usually under five years old. In adults, most of lesions are healed and are interpreted as an indication of childhood episodes of anemia (Lallo et al. 1977; Larsen et al. 1992; Mensforth et al. 1978; Miln er and Smith 1990; Miritoiu 1992; Mittler and Van Gerven 1994; Ribot and Roberts 1996 ; Stodder and Martin 1992; Stuart-Macadam 1985; Walker 1986; Webb 19 95). Long bones, however, are not affected
161 by iron-deficiency anemia (Moseley 1966; So ren et al. 1995). Scur vy, rickets, and bone inflammation may also result in conditions mimicking porotic hyperostosis (Larsen 2000:81). However, with the exception of bone inflammation related to infection, it is unlikely these diseases were affecting th is population since th ey are not commonly observed until much later in history. In addition, there is a possible relationship between porotic hyperostosis and smallpox that should be considered (Peckmann 2003). Dietary deficiencies of vitamins A, B6, B9, B12, C, and the mineral iron (Fe), are also associated with porotic hyperostosis and cribra orbi talia (Walker 2009). The Food and Nutrition Boards (2002) recommendations for the dietary needs for groups per day are presented in Table 7.1. In particular, when addressing the issues of dietary causes for porotic hyperostosis and cribra orbitalia in this population pa rticular attention was given to the dietary needs of children becaus e these conditions de velop early in life. To address the availability of these nutrients, data from archaeological research (Hurcombe 1992b, Lazrus 1999: 126; Piga a nd Porcu 1990; Rowland 2001:41) were used to identify dietary breadth for the nutrients related to por otic hyperostosis and cribra orbitalia. The archaeological data (Table 7.2) show the dietary requirements for A, B6, B9, C, and Fe for individuals under eight y ears of age could be met through a few servings of grains, fruit, or milk products, while B12 needs could be met with two servings of cows milk or one serving (100 g) of meat. However, more analyses of floral and faunal remains are needed to quantif y these data, identify other possible food sources, and understand food processing techniques, which may impact the bodys ability to derive nutrients from these foods. For ex ample, techniques used to process food can impact the bioavailability of micronutrien ts (Holtz and Gibson 2007). Cooking, grinding,
162 Table 7.1. Recommended Dietary Allowances an d Adequate Intakes for iron and vitamins A, B6, B9, B12, and C per day (Food and Nutrition Board 2006) Age Group A (g ) as RAE1 B6 (mg) B9 2 (g) B12 (g) C (mg) Fe (mg) Infants 0-6 m 4003 0.13 653 0.43 403 0.273 7-12 m 5003 0.33 803 0.53 503 11 Children 1-3 y 300 0.5 150 0.9 15 7 4-8 y 400 0.6 200 1.2 25 10 Males 9-13 y 600 1.0 300 1.8 45 8 14-18 y 900 1.3 400 2.4 75 11 19-30 y 900 1.3 400 2.4 90 8 31-50 y 900 1.3 400 2.4 90 8 51-70 y 900 1.7 400 2.45 90 8 >70 y 900 1.7 400 2.45 90 8 Females 9-13 y 600 1.0 300 1.8 45 8 14-18 y 700 1.2 4004 2.4 65 15 19-30 y 700 1.3 4004 2.4 75 18 31-50 y 700 1.3 4004 2.4 75 18 51-70 y 700 1.5 400 2.45 75 8 >70 y 700 1.5 400 2.45 75 8 Pregnancy 14-18 y 750 1.9 6004 2.6 80 27 19-30 y 770 1.9 6004 2.6 85 27 31-50 y 770 1.9 6004 2.6 85 27 Lactation 14-18 y 1200 2.0 500 2.8 115 10 19-30 y 1300 2.0 500 2.8 120 9 31-50 y 1300 2.0 500 2.8 120 9 1 RAE are retinol activity equi valents, which measures the content and activity of vitamin A in foods 2 B9 as naturally-occurring folate 3 Adequate intakes (in the case of healthy, br eastfed infants, this is calculated from mean intake) 4 An additional 400 g per day through supplements is recommended for women of childbearing age and those who are pregnant to prevent neural tube defects during fetal development 5 Because of malabsorption in 10-30% of individuals over age 50, additional consumption of foods or supplements containing B12 is recommended
163 Table 7.2. Nutrient values for foods found in the archaeological reco rd at Neolithic and Bronze-Age sites in Sardinia. All values ar e per 100 g, with the exception of olive oil, which is per tablespoon. From ww w.nal.usda.gov/fnic/foodcomp/search/ Food A (g_RAE) B6 (mg) B9* (g) B12 (g) C (mg) Fe (mg) Almonds 00.143500 03.72 Barley flour/meal 00.39680 02.68 Beef 00.651131.27 01.85 Boar 00.41860.70 0.01.12 Catfish 150.106102.90 0.80.35 Cheese, goat 4860.08040.12 01.88 Cow milk 280.03650.44 00.03 Deer 0ndndnd 04.47 Durum wheat 00.419430 03.52 Fava beans 10.0721040.00 0.31.50 Goat milk 570.04610.07 1.30.05 Grapes 30.08620 10.80.36 Hare 00.34086.51 04.85 Lentils 00.1781810 1.53.33 Mullet 420.490100.25 1.21.41 Olive oil 0000 00.08 Olives 200.00900 0.93.30 Peas, green 380.169650 40.01.47 Pork 290.20611.24 00.64 Sheep 3910.174300 370.70 Tuna 180.981102.19 1.01.60 Wine 00.08410 00.68 soaking, and fermentation can reduce or i nhibit phytates and polyphenols, in turn increasing the ability of nutri ents, such as iron, calcium, and zinc, to be absorbed; however, these results are often dependent upon the species of plan t being processed. Germination and malting, on the other hand, will increase phytate ac tivity. Furthermore, the materials used to process food may also provide a means for introducing nutrients into the diet. For example, Walker (1985) not es that grinding stones, often rich in iron and other minerals, used by Native Ameri cans of the Southwest degrade during food preparation, and small grains are deposited wi thin the processed plan ts and subsequently consumed.
164 At this point, it is also necessary to consider the impact of fava beans ( Vicia faba ) in the diet. The evolution of humans, fava beans, and the malaria para site is considered a textbook example of biocultural adaptation. Fa va beans contain the oxidant isouramil, which has been found to inhibit the growth of malaria parasites in G6PD deficient cells, but not in normal cells (Katz and Schall 1979). It is thought that favism, which affects individuals with G6PD deficiencies when fava beans are eaten or possibly when the pollen of the plant is inhaled, would be select ed against in these environments unless the population was deriving a benefit, such as protection from malaria, by consuming fava beans (Katz and Schall 1986). Furthermore, fava beans ar e generally consumed during periods of malaria outbreaks, and the way th ey are processed impacts their potency and hemolytic effects. Although the results of this study were inc onclusive, an examination of the harvesting and processing of fava bean s by Nuragic populations can provide more information about the relationship of humans, Fabia vica and the malaria parasite. Since the archaeological data show adequate amounts of nutrients in the available food, and conditions such as scurvy appear t ypically much later in history, it seems unlikely the porotic hyperostosis and cribra orbitalia in this populat ion were solely the result of a nutrient deficiency from diet or the result of bone inflammation from infection. Since many factors can influence the formati on of these lesions, in a population it is possible that the presentation of porotic hyperostosis and crib ra orbitalia have different causes in different individuals or a single indi vidual may be subjected to multiple factors that could cause these lesions. However, when considering the dietary evidence, a more plausible interpretation of the porotic hyperost osis and cribra orbitalia in this population would be the result, at least in part, of a hemolytic anemia (Rothschild 2000) or a
165 megaloblastic anemia caused by pathogen lo ad (Kent 1992:17; Stuart-Macadam 1992a; Walker 2009). With an understanding of the an emias afflicting future populations on the island and the malarious nature of Sardinia, it cannot be ru led out that the bony responses observed in this population were the result of inherited hemolytic anemias or chronic hemolytic anemia resulting from malaria. Because data regarding thalassemia in Sardinia were not collected until the mid 1980s (Astolfi et al. 1998:280), it is difficult to estimate the di stribution of this inherited anemia in prehistoric populations. Before couples underwent genetic screening, one out of 250 people born in Sardinia had thalassemia major (WHO 2009c). However, this ratio represents the prevalence of thalassemia after changes in allelic frequencies from gene flow after the Middle Bronze Age, such as the hypothesized spread of the CD39 mutation by the Phoenicians in the first millennium BC. Therefore, the prevalence of thalassemia experienced in the Bronze Age populations shou ld be lower than in modern populations. Using the data provided by the Worl d Health Organization (2009c), the distribution of the alleles, and in turn the predicted frequenc y of heterozygous individuals, can be calculated from the known rate of homozygous recessive individuals by using the Hardy-Weinberg (Hardy 1908) equilibrium equation: p + q =1 if: q = 1/250, or 0.063 p + 0.063 = 1 p = 0.937 p2 = 0.878
166 q2 = 0.004 and: p2 + 2pq +q2 = 1 then: 0.878 + 2(0.937x0.063) + 0.004 = 1 or, in other words: 0.878 + 0.118 + 0.004 = 1 Therefore, at the time of this study, 11.8% of the population would be heterozygous, and 0.4% of the population would be homozygous for thalassemia. If heterozygous individuals have some expression of thal assemia (e.g., anemia), then 12.2% of the population should have some evidence of thalassemic conditions (although, as previously discussed, it is debatable if heterozygous individuals have co nditions and if these conditions would be observable in osseous tissue). However, Cal (2008:42) also observes the distribution of th alassemia and G6PD deficiency are not constant throughout the island; but are significantly inversely related to elevation, with the gene for thalassemia having an average value of 20% and an upper va lue of 36% in populations on the island; however, the data used to ma ke these estimations were not presented. At Serra e Sa Caudeba, 38.4% of th e population had evidence of porotic hyperostosis and cribra orbitalia. Since th is is a childhood condition, and as one ages bony remodeling can obliterate evid ence of these conditions, it is likely this number is an underrepresentation of these responses. If this sample is representative of the population, the rates of porotic hyperostosis and cribra orbitalia would no t likely be attributed solely to thalassemia if using the known rates (WHO 2009c) as a reference (unless the tomb was
167 used for the interment of i ndividuals with thes e conditions, which would result in an overrepresentation), as the difference in the proportions of 0.38 a nd 0.12 is statistically significant, 2 (1, N=1099) = 166.13, p < 0.001. However, when using the higher rates of thalassemia presented by Cal (2008:42), there is not a statistically significant difference, 2 (1, N=199) = 0.15, p = 0.70. Therefore, while it is possible these cond itions are related to an inherited hemolytic anemia, there was no other convincing evid ence of thalassemia observed in this population (Table 7.3). It is also possible a comb ination of inherited hemolytic anemias (such as G6PD deficiency in addition to thalassemi a) could be present in the population for porotic hyperostosis and cr ibra orbitalia to be expressed at these rates. Another possibility is the cause of porotic hyperostosis and cribra orbitalia in these individuals was a para sitic infection. Tognotti (1998:237) notes, before the introduction of quinine, one in eight deaths in Sardinia was the result of malaria. Other research (Le Prince et al. 1916:219) records malarial infection rates to be as high as 50% in the agricultural populations of the United States and 75% of rural populations in the tropics in the early 1900s. From this information, if the para sitic infection affecting this population was malaria, at a given time up to 75% of the population could have been infected with malaria. If these people were living in a malarious environment, unless they were deriving some benefit from an inherite d anemia offering protection from malaria, it is likely that P. falciparum was not contributing to the por otic hyperostosis and cribra orbitalia, as it would have most likely result ed in death before chronic anemia developed. The most likely species affec ting this population would be P. vivax, which, from one mosquito bite, can infect an individual in mu ltiple cycles over several years (Markell et
168 al. 1992:104), thus causing chronic hemolytic an emia. However, it is also necessary to note malaria is not the only infection that causes anemia. 7.4.5 Overall Osteology Results Including Postcranial Bones, Individuals, and Crania The analysis of the remains in this co llection, including the individuals, crania, and other postcranial elements provide clues about the stressors, including diseases, affecting a population. Overall, almost ha lf (46.0%) the paleopathological conditions observed were in the category of porotic hyperost osis and cribra orbitalia. Also of note is the lack of evidence for trauma (fractures a nd dislocations). Arguments have been made that the Nuragic people were warrior pastoralists (Lazrus 1999; Rowland 2001); however, the study of this collection does not provide any evidence supporting this hypothesis. The primary goal of this portion of the re search was to identify conditions related to thalassemia. In addition to using thalassemi a as an indirect indicator of malaria, it was previously noted the comorbidity of malaria an d tuberculosis results in a complex disease process (Table 7.3). Malaria makes an individual more susceptible to tuberculosis, even if they do not have symptoms of a malarial in fection (Enwere et al. 1999). In individuals affected with chronic malaria, the disease process is lengthened and can result in presentation in the skeleton. Therefore, in a population infected with malaria and tuberculosis, it is likely there will be evidence of tuberculosis in the skeletal remains of a population. On the other hand, syphilis and malari a have an antagonistic relationship; in an area with endemic malaria, there would be little to no evidence of syphilis (Fraser 1998). However, it is debatable that this disease was present during the Bronze Age (Ortner and Putschar 2003:297299). In the skeleton, tuberculosis presents primarily
169 Table 7.3. The number of observations of each pathological condition observed, and the association of thes e conditions with thalassemia and tuberculosis (after Ortner and Putschar 2003) Pathology Number of Observations Thalassemia Tuberculosis Porotic hyperostosis 63 + Lytic lesion, vertebra 22 + Cribra orbitalia 17 + Arthritis, femur 10 + Angling of vertebral body (possible compression fracture) 7 + + Osteophytes, vertebral. Fragments 6 + Schmorls node 6 Abnormal bone loss and formation (possible treponematosis) 4 Arthritis, humerus 4 + Hyperostosis, long bone fragment 4 Myositis ossificans 4 Endocranial lesions 3 + Arthritis, ulna 2 + Arthritis, vert. 2 + Lesion, sternum 2 + Lytic lesion, humerus 2 + Porosity, cervical vertebra. 2 + Syndesmophytes, vertebral fragment 2 Ankylosis and kyphosis, cervical vertebrae 1 + Arthritis, acetabulum 1 + Arthritis, humerus 1 + Arthritis, phalanx 1 Arthritis, tibiae 1 + Cloaca, fibula 1 Cloaca, ulna 1 Dislocation, glenoid fossa 1 Fracture, occipital 1 Hyperostosis, ulna 1 Hypoplasia, mastoid processes (bilateral) 1 ? ? Lesion, navicular (foot) 1 + Lesion, ulna 1 + Lytic lesion, calcaneus 1 + Meningeal reaction with granular impressions 1 ++ Multifocal lesions, ilium 1 + Necrosis, femur 1 +
170 Table 7.3. The number of observations of each pathological condition observed, and the association of thes e conditions with thalassemia and tuberculosis (after Ortner and Putschar 2003) (continued) Pathology Number of Observations Thalassemia Tuberculosis Periostitis, femur 1 Periostitis, rib 1 + Cortical destruction and trab ecular bone with right angle crossings, ribs 0 + Destructive remodeling, ribs 0 + Dwarfism 0 + Enlarged nutrient foramina, phalanges 0 + Flask-shaped deformities, long bones 0 + Harris lines 0 + Honey-comb pattern of subperiosteal bone 0 + Kyphosis of thoracic or lumbar vertebrae 0 + Lesions in tubular bones of hands and feet 0 + Malocclusion 0 (u) + Maxillary prognathism 0 (u) + Narrow nasal cavity 0 (u) + Obliterated maxillary sinus 0 (u) + Prominent mandibular incisors 0 (u) + Prominent zygomatic bones 0 (u) + Pronounced vertical growth of maxilla 0 (u) + Protrusive maxillae 0 (u) + Severe changes in cortical bone (fissures, thinning, mineralization defects) 0 + Underbite 0 (u) Vertebrae with increased width, decreased height, and cupping 0 + Widening of medullary cavity 0 + u = condition of remains did not allow for the observance of this condition in this collection + = associated with the condition ++ = reliable indicator of the condition = not associated with the condition ? = is not known if condition is related to the disease
171 in lesions on the thoracic and lumbar verteb rae, the sternum, and on the pleural surfaces of the ribs (in the form of resorption and proliferation of bone cells) (Buikstra 1981a; Larsen 1997; Steinbock 1976). Lesions also a ppear on the long bone metaphyses in sub adults (Hopewell 1994, Ortner and Putschar 2003, Thijn and Steensma 1990). However, any bone or joint can be invol ved (Berney et al. 1972). The di sease process results in the destruction of cancellous bone, lo ss of bone mass, with the possi bility of vertebral bodies collapsing resulting in severe kyphosis. Sin ce rib lesions could be the result of other pulmonary disease, their presence alone should not be interpreted as tuberculosis (Pfeiffer 1991; Roberts et al. 1994). Tube rculosis has been traced to 3000 BC in the Old World (Daniel et al. 1994; Steinbock 1976). 7.5 Discussion of Malarial aDNA, Pf HRP II, and Hemozoin Analysis from Serra e Sa Caudeba Tomb B Ancient DNA analysis has value in kinship analysis, sex identification, and palaeodisease (Brown 2000:462-469). The examin ation of disease th rough the use of aDNA studies provides an understanding of the way a pathogen evolves over time (Hummel 2003:5). However, the analysis of human remains for aDNA is not without its problems. To begin with, without replication (ideally at anothe r laboratory), the results of aDNA are provisional; however, the costs associ ated with replicati on are prohibitive, so this is not usually done (O Rourke et al. 2000:226). In this study, it is likely that the initial results were false positive because of the sensitivity of the test (personal comm unication with David Sullivan, Jr. 2009) or contamination with contemporary DNA, such as airborne particles in the laboratory
172 (Hummel 2003:66). These negative results do no t necessarily mean that this population was not afflicted with malaria; however, withou t positive test results, it is not possible to provide evidence of the presen ce of malaria. For example, some factors which may contribute to the negative resu lts of the aDNA, protein, and hemozoin tests include the length of the storage of the bone at room temperature, the extr action techniques and protocols, the choice of markers and prim er design, inhibiting substances, or PCR parameters (Hummel 2003:7,72). In addition, the number of bones selected, if they were from a population with malaria (using an estima te of a 40% infection rate), would have statistically included four samp les from infected individuals, yet the overall detection rate for the tests performed was not taken into account when sampling. 7.6 Chapter Summary This research demonstrates the value of the use of multiple lines of evidence. This included the consideration of data from various sources (e.g., ecological, historic, anthropological, and medical), and interdis ciplinary collaboration. By approaching a research question in this manner, rather than focusing on one line of evidence, there is a greater amount of information considered, and additional methods can be used when testing a hypothesis. For exam ple, different conclusions would have been made by only considering the results of the osteological study without other arch aeological data (e.g., those resulting in dietary information) or the data collected during the eradication of malaria on Sardinia. While the most desirable collection with which to work is one composed of complete skeletons, it does not mean colle ctions with highly-fragmented, commingled
173 remains lack research value. An analysis of fragmented remains can result in the identification of patterns of skeletal responses that merit additional research, for example considering aDNA analysis for the suspect ed presence of a trait or pathogen, or identifying remains for histological or radi ographic analysis, such as in the case of porotic hyperostosis. Porotic hyperostosis and cribra orbitalia are regarded as an indicator of childhood anemia, and the bones of adults may have obl iterated evidence of this condition, it is likely the prevalence of the condition causi ng these responses was higher than observed, which would be consistent with the preval ence of malarial inf ections or inherited hemolytic anemias in some regions of Sardinia before the disease process was completely understood. Since there are many thoughts regard ing the cause of porotic hyperostosis and cribra orbitalia, from iron deficiency anem ia to pernicious and hemolytic anemias, it is not possible to attribute th is condition to malaria or an inherited hemolytic anemia, such as thalassemia. Further examination of the etiology of porotic hyperostosis and cribra orbitalia at Serra e Sa Caudeba, th rough the use of radiologi cal or histological techniques on a representative sample from this population, is necessary to understand the cause or causes of these lesions in this population. Although no pattern of bony responses consistent with th alassemia major was observed, it is possible that other inherited hemolytic anemias, such as G6PD deficiency or even thalassemia minor, could cause porotic hyperostosis and cribra orbitali a without other skeletal responses. Since individuals with thalassemia major need medical care to survive, it is possible only those with the milder expression of the disease (e.g., thalassemia minor) survived long enough for their bones to respond to the condition. In addition, the examination of the bones also
174 revealed a pattern of pathologies consistent with tuberculosis which would be expected if the population was exposed to both pathogens. Because the osteological examination resu lted in the identification of conditions consistent with those expected in a population infected with malaria, yet could not result in a definitive diagnosis, further testi ng was necessary. This was done through the examination of aDNA, immunoassay, and hemo zoin detection. The overall results of these tests were negative. However, because of the condition of the remains and the length of time since they were excavate d, the likelihood of extracting malaria DNA or protein is low. In addition, while hemozoin is a good diagnostic tool for malaria, it has not successfully been identified in human remains from an archaeological context. Hemozoin isolation techniques may need to be modified for applications in bioarchaeology in order to produce positive resu lts. The detection of hemozoin and other biocrystals would be a valuable tool for understanding disease and human behavior in prehistory. When testing the research hypothesis (m alaria was present during the Bronze Age), the condition of the remain s contributes to the possibility of type I (rejecting a null hypothesis when it is true, or a false positiv e) and type II (failing to reject a null hypothesis when it is not true, or a false negative) errors. For example, the instability of aDNA could create negative test results when, in fact, the individuals were infected. This could lead to failing to reject the null hypothesis, resulting in a type II error. On the other hand, the nature of this research, if produc ing negative results, does not conclusively allow for the null hypothesis to fail to be rej ected (that is absen ce of evidence is not evidence of absence).
175 However, when considering the data in its entirety, it is not unreasonable to suggest that malaria was present during the Bronze Age in Sardinia. The gross osteological examination and interpreta tion of dietary evidence (isotopic and archaeological) suggest that the porotic hypero stosis and cribra orbitalia observed were most likely the result of a parasitic infecti on or an inherited anem ia, of which the ones most commonly observed in Sardinia are hemo lytic in nature. Since the recent genetic testing from contemporaneous Egyptian mummi es (Hawass et al. 2010) provides further evidence of falciparum malaria during the late r portion of the time pe riod in question (at minimum), it is quite possible that trade ne tworks in the Eastern Mediterranean including Greece, Cyprus, and Egypt provided a means for the Plasmodium sp. to be transmitted to Sardinia during the Bronze Age.
176 Chapter 8: Conclusions This research is beneficial because it provides a better understanding of life during the Bronze Age in the central Mediterranean, with a specific focus on the Nuragic population of Sardinia. These findings range from the specific, such as the health of the population studied, to the broad, such as the identification of possible malarious conditions and biocultural adaptations to a malarious environment. The examination of archaeological data for evidence of contact with other populations gives a perspective necessary to interpret the hea lth and adaptations of prehistoric Sardinians. In addition, the use of methods from other disciplines to study human remains from archaeological contexts provided additional lines of evid ence for this research. For instance, the application of a new diagnostic technique for hemozoin provided a novel way to approach the challenges of detecting malari a in human remains, while the use of an immunological technique allowed for the testing of the presence of PfHRP II. Further controlled studies with these tests could result in new methods for analyzing human remains. 8.1 Major Findings While there was no conclusive evidence of malaria in this population in the form of aDNA, protein, or hemozoin, the data (i.e., environmental reconstruction and the technological and agricultural demands placed upon the environment as discussed in
177 Chapter 4) suggest conditions were favorable for malaria parasites to be transmitted to humans. Although speculations about the nura ghi providing protecti on from a malarious environment have been made (Flumene in Brotzu 1934; Sallares 2002:91-92), research (Aitken 1953:320-325) found female mosquitoes that would transmit malaria resting in these structures. Without further cultural modi fications to these stru ctures, it is unlikely the nuraghi offered any prot ection from malaria. It is more likely Nuragic water management practices, such as the construc tion of well temples or placement of wells within other Nuragic structures, provided c over from the sun and lower temperatures in the springs, thus creating an inhospitable environment for mosquitoes and parasites (see Aitken 1953:320-325). Likewise, the analysis of the osteologi cal collection supports the possibility that the Nuragic people were infected with mala ria. High rates of porotic hyperostosis and cribra orbitalia were observed. It is suggested that these bony responses are not related to dietary insufficiencies; however, radiologica l or histological anal yses are required to understand the cause of these conditions in th ese individuals. Alt hough the specific cause of the lesions observed in the cr ania is indeterminate, it could be related to pathogen load (consistent with but not limited to malari a) or a genetic hemolytic anemia (e.g., thalassemia or G6PD deficiency); however, th e specific pattern of os teological conditions indicative of thalassemia was not observed, and other types of anemia may be inherited. Additional lesions observed throughout the co llection are suggestive of tuberculosis infections with sclerosis, wh ich would be expected in populations with malaria (Collari 1932:324).
178 Finally, although not related to the testing of the hypothesis of this study, the minimal amount of trauma observed on the remains is noteworthy. Nuragic people are often referred to as warrior pastoralists (L azrus 1999; Rowland 2001), yet as previously stated, this research does not support that hypo thesis. While it is possible individuals who were injured or killed during battles were interred elsewher e, no evidence of separate burials for warriors has been discovered. 8.2 Strengths and Limitations The strengths of this research include the use of multiple lines of evidence, interdisciplinary collaboration, and a synthe tic review of literature. For example, by examining findings from the Sardinian Projec t (Aitken 1953), other historical documents, archaeological evidence, biocultural adapta tions to malaria, and the osteological evidence, I could not only test the hypothesis, but determine if conditions were conducive for malaria during the Bronze Age. The exam ination of the skelet al remains included using techniques associated with physical anthropology, as well as those used by the medical community (e.g., Western blots and he mozoin testing with laser desorption mass spectrometry). This collaborative approach is useful for providing the most advantageous amount of evidence when testing the hypothesi s, especially when addressing a disease such as malaria that does not result in a specifi c bony response or pa ttern of responses. The inclusion of archaeological evidence with the biological data is necessary to understand and identify possible biocultural adap tations to malaria. While the presence of malaria in Sardinia during the Bronze Age ha s been considered (Brown 1984; Cao et al. 1989:309-312; Sallares et al. 2004:327; Sa nna et al. 1997:295; Sondaar 1998), this
179 research examines the evidence comprehensiv ely and applies anthropological techniques to test a hypothesis concerning malaria infect ions on this island during the Middle Bronze Age. This research was not without its limita tions. For example, the small sample size used for aDNA, hemozoin, and protein analysis as well as the fragmented nature of the collection and the focus of the study contribute to the lim itations of this research. Although the remains were fragmented, this collection was considered valuable for testing this hypothesis because of the location of the tomb on the Campidano Plain, the large number of individuals interred at th is tomb, and the presence of a questionable burial feature containing juve nile remains, which may have been indicative of an epidemic. While the condition of the remains limited some of the research that could be conducted (e.g., the reconstruction of skulls to obtain craniometric measurements useful in identifying possible thala ssemic individuals or the abil ity to perform differential diagnoses on complete skeletons ), the larg e quantity of cranial fragments provided the necessary data for estimating rates of porot ic hyperostosis and cribra orbitalia, both required for addressing the issue of acquire d and inherited hemolytic anemias. As previously discussed, examination of these lesions using radiological and histological techniques would provide inform ation that is necessary to understand the cau se of these bony responses. In addition, it was possible to infer other conditions and diseases from the large number of fragmented remains of the entire population (with the caveat that complete individuals were not available). Th e problems inherent with aDNA analysis and the preservation of DNA over millennia provide challenges in this type of research. This analysis demonstrated that that future resear ch would benefit from testing a larger sample
180 for aDNA, hemozoin, and protein (e.g., usi ng a sample of at least 27, assuming a 40% infection rate and a 10% dete ction rate, would be necessary to detect one infection). Finally, although hemozoin detection by laser desorption mass spectrometry is used to diagnose malaria infections in human blood, it has yet to be detected in human remains, in particular bone, from an arch aeological context. Th e literature review for this research revealed no (published) indicati on of archaeologists attempting to detect biocrystals, such as hemozoin, in archaeological remains. Recent advances in the detection of biocrystals, which are likely to be more durable th an DNA or proteins, through mass spectrometry give us the potential to unde rstand the life and he alth of prehistoric populations better. Although these challenges result in limitati ons specific to this research, additional investigation into thes e issues can provide a valuable foundation for future research. 8.3 Contributions and Implications This research provides information resu lting in a more complete understanding of Nuragic life. Because this study was only one component of the analysis, which is concurrently being written, of the populati on at Tomb B of Serra e Sa Caudeba there is much to be learned from this population in conjunction with what has been presented here. For example, estimates on stature and the results of stable isotope analysis, especially when compared with other populatio ns, can give insight into the evolution of humans in island or malarial environments a nd dietary information, both of which can be of use when considering the possibility of ende mic malaria. In the absence of evidence of malaria, the gross osteological examination gi ves data pertaining to the health of these people. For instance, the indications of ch ildhood anemia or infections resulting in
181 porotic hyperostosis and cribra orbitalia, possible tuberculosis infections, and the lack of trauma from violence cannot be ignored. The results of this examination provide further evidence of agricultural activity during the Middle Bronze Age in Sardinia. The dissertation also discusses the impor tance of using the theories and methods of the social sciences (Oakes 1991; Williams et al. 2002), in part icular anthropology, to understand the evolution of and adaptations to malaria. By doing so, more information is available to fight this disease. For ex ample, by identifying and studying malaria aDNA present in human bones from various times, the evolution of the parasite can be better understood, and its DNA sequenced, both of wh ich are important steps to developing malaria vaccines and new, non-resistant drugs and pesticides (Baleta 2009). In addition, effective biocultural adaptations can be iden tified, which would be of use for civil and environmental engineers working in countries where malaria is endemic. For instance, features from wells, channeling systems, a nd water storage methods that prevent the growth mosquitoes can be incorporated into the design of new water management techniques in malarious regions. 8.4 Directions for Future Research Researching malaria through the use of archaeological data, DNA, and human remains can include many approaches. These approaches can be categorized into the development of methods to detect malaria in human remains, sequencing malaria DNA from these remains when possible, the examination of human remains from other populations, and the synthesis of research fr om various disciplines to gain a better understanding of the coevolution of Plasmodium sp. and humans. It is also important to
182 iterate that the examination of the remains in this study was made with the intent of testing a specific hypothesis; a complete osteological examination and recording of all elements was not made. There are many benefits to examining the remains further. In particular, radiographic and histological studies can provide more insi ght into the nature of the porotic hyperostosis and cribra orbitalia observed. These rema ins may also be studied to test other hypotheses, such as those pertaini ng to the demography, heal th, and diet of this population. In addition, it would be useful to obtain radiocarbon da tes so the use and deposition of the tomb could be better understood, while expanding the chronological data available for this island. Detecting malaria in human remains i nvolves more than an osteological examination of skeletal material. Because there is no specific pa ttern of bony responses indicative of malaria, other techniques must be used. At the present, the most convincing diagnostic methods include aDNA analysis and immunological assays. Both of these procedures require well-preserved remains. New techniques for the detection of hemozoin, which is considered insoluble, coul d be useful in the id entification of malaria infections in prehistoric populations. However, because hemozoin has never been successfully isolated from remains from an archaeological context, controlled experiments must be conducted to establish protocols for detecting this biocrystal in osseous or mummified tissue. The detecti on of hemozoin in human remains would provide an important tool for anthropologists researching the health and cultures of past populations, especially when other detection methods fail. In addition, because hemozoin is insoluble, the presence of this biocrystal could se rve as a screening method for
183 identifying candidates for aDNA analysis. Th e sequencing of ancient malarial DNA is necessary to understand the evol ution of this parasite, whic h is useful for developing nonresistant drugs and vaccines (Hartl et al. 2002). Since no conclusive genetic evidence of malaria was detected in this population, future research producing micr obiological data consistent with malaria infections is needed before evidence of its presence can be well supported. Likewise, the future excavation and proper collection of huma n remains for human aDNA analysis is necessary to test for genetic evidence of conditions such as thalassemia or G6PD deficiency. Furthermore, remains from other populations could be examined using similar methods. This could include studying better-pre served prehistoric remains from Sardinia in a similar manner, or it could consist of examining remains from a historic period when malaria was described on Sardinia (e.g., the Roman period). Analysis of the infant cremations at Tharros may also provide valuable information. Examining osteological data (gross, radiologic, and histologic) and analyzing the bones for aDNA, Pf HRP II, and hemozoin from individuals known to have malaria is a necessary step for making a differential diagnosis of this disease (B uikstra 1976). Finally, to understand the relationship between malaria, humans, and mosquitoes, a synthesis and dissemination of interdiscipl inary datasets is needed. By synthesizing decades of research from various disciplines and creating and publishing digital datasets, researchers studying and combating malaria th roughout the world would be able to access data they might not be able to obtain otherw ise. The compilation of data from the various interpretations of popula tion genetic studies of Plasmodium sp., Anopheles sp. and inherited hemolytic anemias can also result in the creation of geodata bases so spatial and
184 temporal investigations can be made. In addition, a comprehens ive, cross-cultural analysis of individuals living in malarious environments would be beneficial to identifying adaptations to this disease a nd should be included with these data. 8.5 Summary and Conclusions The primary purpose of this research was to test the hypothesis that malaria was present on Sardinia during the Middle Bronze Age. Although malaria was not detected in the aDNA, immunology, and hemozoin anal yses, the review of literature and interpretation of collected oste ological data indicate ecological and social conditions were favorable for the transmission of this parasi te. The possibility malaria influenced the Nuragic people, both genetically and culturally, cannot be ruled out and is still very reasonable. The secondary purpose of this research was, if evidence supported the argument for a malarious environment during the Midd le Bronze Age, to identify biocultural adaptations to the disease. In particular, attention was paid to cultural features long speculated to provide protection from malaria, specifically the nuraghi. A review of the literature did not support the hypo thesis that these towers (w ithout further modification) were an adaptation to malaria, but more li kely defensive or aggrandizing structures representing social capital and possibly signi fying land ownership. However, the review did show that Nuragic wells were not conduciv e to sustaining malaria, and the building of these structures (and possibly the use of ch annels) and rituals and behaviors associated with water may have helped maintain the health of the associ ated village population. According to the osteological examinati on, approximately 40% of the population had
185 evidence of porotic hyperostosis and cribra or bitalia, which was most likely related to the pathogen load or an inherited, most probably hemolytic, anemia. Therefore, it is also possible genetic adaptations to mala ria were present at this time. Although half of the world is at risk for ma laria, this disease is mainly present in regions with high rates of poverty and conflict, and therefore receives a disproportionately low level of attention, mainly from the medical disciplines (World Health Organization 2009d). However, social sc ientists have the ability to research the problem of malaria transmission by including the role of human behavior and adaptation (Williams et al. 2002:251). The results of anthropological research, such as this archaeological study, can be applied and incor porated in the development of strategies used to combat disease in contemporary populations.
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231 Appendix A: Artifacts fr om Serra e Sa Caudeba During the excavations at Serra e Sa Caudeba, few artifacts were recovered from Tomb A because the site was disturbed. Alt hough Tomb B remained intact, there were still few artifacts associated with the remains. These artifacts in cluded ceramic vessels, beads, and a bronze dagger. The stylistic attribut es of these artifacts suggest that they are from the Middle Bronze Age or Nuragic I period (ca. 1600-1300 BC). The glaze on the ceramics is indicative of possible Mycen aean contact and influence (personal communication with Mauro Perra 2008). All photographs in th is section were taken by the author.
232 Appendix A: Artifacts fr om Serra e Sa Caudeba (Continued) Figure A.1. Incised truncated terracotta cup, or tazza from Tomb A Figure A.2. Clay beads from Tomb A
233 Appendix A: Artifacts fr om Serra e Sa Caudeba (Continued) Figure A.3. Bronze dagger blade from Tomb A Figure A.4. Hemispheric terracotta cup from Tomb A
234 Appendix A: Artifacts fr om Serra e Sa Caudeba (Continued) Figure A.5. Terracotta globular urn from Tomb B Figure A.6. Clay beads from Tomb B
235 Appendix A: Artifacts fr om Serra e Sa Caudeba (Continued) Figure A.7. Terracotta globular urn from Tomb B Figure A.8. Terracotta sherds and ur n lid (or plate) from Tomb B
236 Appendix B: Data Collection Protocol The data collection protocol for this disse rtation was created to focus on skeletal elements useful for observing conditions relate d to anemia and thalassemia. However, the importance of standardizing the data was consid ered. The standardization of these data is necessary to compare individuals or collections in the future. Therefore, the skeletal remains were observed, described, and recorded in accordance with Buikstra and Ubelaker (1994) and Kimmerle (2 009). Inventories of individual skeletons, sex and age estimations, paleopathologies, and taphonomi c responses were completed using the following guidelines.
237 Appendix B: Data Collection Protocol (Continued) Protocol for the Inventor y of Individual Skeletons (after Buikstra and Ubelaker 1994:7, Attachment 1: Chapter 2) Individual: Context: Date: Bone Types Method Code Cranial and Postcranial Bones other than Cervical Vertebrae 3-6; Thoracic Vertebrae 1-9; Ribs 3-10; Carpals, Metacarpals; Tarsals (except Talus and Calcaneus); Hand and Foot Long bone diaphyses divided into proximal, middle, and distal thirds; each section and epiphyses/articular surfaces are recorded separately Blank = missing 1 = >75% present complete 2 = 25% 75% present = partial 3 = <25% present = poor Cervical Vertebrae 3-6; Thoracic Vertebrae 1-9; Ribs 3-10 Bodies recorded separate from neural arches; total number of observable units recorded (C3-C6; T1-T9; left and right ribs 3-10 separately); total count of complete units by code Bodies/Centra Complete = >75% present Neural Arches Complete = at least two observable articular facets Ribs Complete = head and neck present Carpals, Metacarpals, Tarsals (other than Talus and Calcaneus); Metatarsals, Hand and Foot Phalanges Left and right elements separated (other than phalanges) and counted; Number of bones present, with a count of complete (>75% present) specimens Fragments not assigned to one of the above categories Sorted into the categories of skull, limb long bones, vertebrae, hand/foot, clavicle/scapula, os coxae, miscellaneous postcranial, and unidentifiable Report by count or weight
238 Appendix B: Data Collection Protocol (Continued) Protocol for the Inventory of Individual Skeletons (Continued) (after Buikstra and Ubelaker 1994:7, Attachment 1: Chapter 2) Individual: Context: Date: Cranial Elements Left Right Left Right Frontal Sphenoid Parietal Zygomatic Occipital Maxilla Temporal Palatine TMJ Mandible Postcranial Elements Left Right Left Right Clavicle Os Coxae Scapula body Auricular Surface Glenoid f. Ilium Patella Ischium Sacrum Pubis Acetabulum Vertebrae (Individual) Vertebrae (Grouped) # Present/ # Complete Centrum Neural Arch Centra Neural Arches C1 C3-6 C2 T1-T9 C7 Manubrium Body T10 Sternum T11 Ribs (Grouped) # Present/# Complete T12 L R U L1 3-10 L2 Ribs (Individual) L3 1st L4 2nd L5 11th & 12th
239 Appendix B: Data Collection Protocol (Continued) Protocol for the Inventory of Individual Skeletons (Continued) (after Buikstra and Ubelaker 1994:7, Attachment 1: Chapter 2) Individual: Context: Date: Long Bones Proximal Epiphysis Proximal Third Middle Third Distal Third Distal Epiphysis Left Humerus Right Humerus Left Radius Right Radius Left Ulna Right Ulna Left Femur Right Femur Left Tibia Right Tibia Left Fibula Right Fibula Left Talus Right Talus Left Calcaneus Right Calcaneus Hand (# Present/# Complete) Foot (# Present/# Complete) L R U L R U #Carpals #Tarsals #Metacarpals #Metatarsals #Phalanges #Phalanges
240 Appendix B: Data Collection Protocol (Continued) Protocol for Adult Sex Estimation (after Buikstra and Ubelak er 1994:16-20; Kimmerle 2009) Individual: Context: Date: Os Coxae Morphology: Attribute Left Right Ventral Arch* Subpubic Concavity* Ischiopubic Ramus Ridge* Greater Sciatic Notch** Preauricular Sulcus*** *blank = unobservable, 1 = female, 2 = ambiguous, 3 = male ** 0 = unobservable, 1-5 based upon width, most feminine (wide) to most masculine (narrow) *** 0 = absent, 1 = wide and deep, 2 = wi de and shallow, 3 = well defined but narrow, 4 = narrow, shallow, and smooth-walled depression Feature F M Superior Inlet Oval-shaped Heart-Shaped Pubic Bone Rectangular Triangular Ventral Arc Present Absent Subpubic Concavity Present Absent Ischio-pubic Rami (medial surface) Ridge Present Broad, Flat Auricular Surface Raised Flat Scars of Parturition Present Absent Length of Pubis x 100 Length of Ischium Estimated Sex from Os Coxae (1-5): _______ (1 = most feminine, 5 = most masculine)
241 Appendix B: Data Collection Protocol (Continued) Protocol for Adult Sex Estimation (Continued) (after Buikstra and Ubelak er 1994:16-20; Kimmerle 2009) Cranial Morphology: Feature L M R Glabella n/a n/a Supraorbital Margin n/a External Nuchal Protuberance n/a n/a Mastoid Process n/a Mental Eminence n/a n/a (features are scored 1-5; 1 most feminine, 5 most masculine) Feature F M Forehead Vertical Retreating Temporal Crest Absent/SlightDeveloped Nuchal Lines Absent/SlightDeveloped Suprameatal CrestAbsent/SlightDeveloped Gonial Angle > 125 Close to 90 Estimated Sex from Cranium (1-5): _____ (1 = most feminine, 5 = most masculine)
242 Appendix B: Data Collection Protocol (Continued) Protocol for Adult Age Estimation (after Buikstra and Ubelak er 1994:16-20; Kimmerle 2009) Individual: Context: Date: Cranial Suture Closure: External Cranial Vault Score Palate Score Midlambdoidal Incisive Lambda Anterior Medial Palatine Obelion Posterior Medial Palatine Anterior Sagittal Transverse Palatine Bregma Midcoronal Interior Cranial Vault Pterion Sagittal Sphenofrontal Left Lambdoidal Interior Sphenofrontal Left Coronal Superior Sphenofrontal (closure scored: blank = unobservable, 0 = open, 1 = minimal, 2 = significant, 3 = complete) Postcranial Observations: Feature Left RightMean Age and Age Interval Pubic Symphysis Todd (1-10) Suchey-Brooks (1-6) Auricular Surface Lovejoy (1-8) Sternal Rib End Iscan et al. (0-8)
243 Appendix B: Data Collection Protocol (Continued) Protocol for Adult Age Estimation (Continued) (after Buikstra and Ubelak er 1994:16-20; Kimmerle 2009) Estimated Age: Young Adult (20-35 years) ___________ Middle Adult (35-50 years) ___________ Old Adult (50+ years) ___________ Comments:
244 Appendix B: Data Collection Protocol (Continued) Protocol for Juvenile Age Estimation (after Buikstra and Ubelak er 1994:16-20; Kimmerle 2009) Individual: Context: Date: Epiphyseal Union (Krogman and Iscan 1986): Bone Epiphysis Stage of Union Cervical Vertebrae Superior Inferior Thoracic VertebraeSuperior Inferior Lumbar Vertebrae Superior Inferior L R Scapula Coracoid Acromion Clavicle Sternal Humerus Head Distal Medial epicondyle Radius Proximal Distal Ulna Proximal Distal Os Coxae Iliac crest Ischial tuberosity Femur Head Greater trochanter Lesser trochanter Distal Tibia Proximal Distal Fibula Proximal Distal (union scored as: blank = unobservable, 1 = partial union, 2 = complete union)
245 Appendix B: Data Collection Protocol (Continued) Protocol for Juvenile Ag e Estimation (Continued) (after Buikstra and Ubelak er 1994:16-20; Kimmerle 2009) Primary Ossification Centers: Bone Area of Union Extent Os Coxae Ilium-pubis Ischium-pubis Ischium-ilium Sacral Segments 1-2 2-3 3-4 4-5 Cervical Vertebrae Neural arches to each other Neural arches to centrum Thoracic VertebraeNeural arches to each other Neural arches to centrum Lumbar Vertebrae Neural arches to each other Neural arches to centrum Cranium Spheno-occipital synchondrosis Occipital Lateral part to squama Basilar part to lateral part (union scored as: blank = unobservable, 1 = partial union, 2 = complete union) Estimate of Chronological Age from Postcranial Skeleton: Fetal b-5 5-10 10-15 15-20 20+
246 Appendix B: Data Collection Protocol (Continued) Protocol for Juvenile Ag e Estimation (Continued) (after Buikstra and Ubelak er 1994:16-20; Kimmerle 2009) Dental Development (Moores et al. 1963): Deciduous: Max Inc. 1 Inc. 2 Canine Molar 1 Molar 2 Man Inc. 1 Inc. 2 Canine Molar 1 Molar 2 Permanent: Max I1 I2 C PM1 PM2 M 1 M2 M3 Man I1 I2 C PM1 PM2 M 1 M2 M3 Age Estimation from Dental Eruption (Ubelaker 1989): _______ Comments:
247 Appendix B: Data Collection Protocol (Continued) Protocol for Recording Pale opathologies (Continued) (after Buikstra and Ubelak er 1994:16-20; Kimmerle 2009) Individual: Context: Date: Bone: Side: Section: Aspect: Pathology Code*: Obs1: Obs2: Obs3: Obs4: Obs5: Obs6: Obs7: Obs8: Obs9: Obs10: Comments and additional observations: For pathology codes, see Buikst ra and Ubelaker (1994:114-115).
248 Appendix B: Data Collection Protocol (Continued) Protocol for Taphonomy Recording (after Buikstra and Ubelak er 1994:16-20; Kimmerle 2009) Individual: Context: Date: Alteration Type LocationPhotograph #Additional Information
249 Appendix B: Data Collection Protocol (Continued) Protocol for Taphonomy Recording (Continued) (after Buikstra and Ubelak er 1994:16-20; Kimmerle 2009) Types of alterations recorded: 1. Weathering (include bone identifica tion, photograph numbers, and degree of weathering) 2. Discoloration (include bone identi fication, location, photograph numbers, Munsell color system code for normal and discolored bone) 3. Polish (include bone identification and photograph number) 4. Cutmarks (include bone identifi cation, location, photograph number, number of cuts, average cut length, range of cut lengths) 5. Animal Gnawing (include bone id entification, location, photograph number, number of paired grooves or incisions, specify rodent vs. carnivore gnawing) 6. Other Cultural Modification, including creation of artifacts (include bone identification and photograph number) 7. Burned Bone* (include colors of bones, percentage /weights of bone affected by color, surface texture, bone deformity, and surfaces shielded) For details on burned bone recording, refer to Buik stra and Ubelaker (1994: Attachment 23).
250 Appendix C: Raw Data The following raw data were taken from the observations made using the protocol in Appendix B. Inventories of the individua l skeletons were made. Pathologies observed on the individuals, cranial elements, and i ndividual elements were recorded. Taphonomy is discussed in Chapter Six.
251 Appendix C: Raw Data (Continued) Inventory of Individual Skeletons Individual a, Serra e Sa Caudeba Tomb B: Cranial Elements Left Right Left Right Frontal Sphenoid Parietal Zygomatic Occipital Maxilla Temporal Palatine TMJ Mandible Postcranial Elements Left Right Left Right Clavicle 2 2 Os Coxae Scapula body Auricular Surface Glenoid f. Ilium 3* Patella 1 1 Ischium Sacrum 3 3 Pubis Acetabulum Vertebrae (Individual) Vertebrae (Grouped) # Present/ # Complete Centrum Neural Arch Centra Neural Arches C1 C3-6 6/0** 28*** C2 T1-T9 5/0** C7 Manubrium Body T10 Sternum 3 T11 Ribs (Grouped) # Present/# Complete T12 L R U L1 3-10 27** L2 Ribs (Individual) L3 1st L4 2nd L5 11th & 12th
252 Appendix C: Raw Data (Continued) Inventory of Individual Skeletons (Continued) Individual a, Serra e Sa Caudeba Tomb B (Continued): Long Bones Proximal Epiphysis Proximal Third Middle Third Distal Third Distal Epiphysis Left Humerus 1 1 1 1 Right Humerus 1 1 1 1 Left Radius 1 1 1 1 1 Right Radius 1 1 1 1 1 Left Ulna 1 1 1 1 Right Ulna 1 1 1 1 Left Femur 1 1 1 1 1 Right Femur 1 1 1 1 1 Left Tibia 3 Right Tibia 1 1 1 1 Left Fibula 1 1 1 1 1 Right Fibula Left Talus Right Talus Left Calcaneus 1 Right Calcaneus Hand (# Present/# Complete) Foot (# Present/# Complete) L R U L R U #Carpals #Tarsals #Metacarpals #Metatarsals #Phalanges 6/0 #Phalanges 1/0 = unsided ** = highly fragmented, could not identify to specific bone *** = highly fragmented, counted and grouped
253 Appendix C: Raw Data (Continued) Inventory of Individual Skeletons (Continued) Individual b, Se rra e Sa Caudeba Tomb B: Cranial Elements Left Right Left Right Frontal Sphenoid Parietal Zygomatic Occipital Maxilla Temporal Palatine TMJ Mandible Postcranial Elements Left Right Left Right Clavicle 3* Os Coxae Scapula body Auricular Surface Glenoid f. Ilium 3* Patella Ischium Sacrum Pubis Acetabulum Vertebrae (Individual) Vertebrae (Grouped) # Present/ # Complete Centrum Neural Arch Centra Neural Arches C1 C3-6 2*** C2 T1-T9 4/0** C7 Manubrium Body T10 Sternum T11 Ribs (Grouped) # Present/# Complete T12 L R U L1 3-10 5** L2 Ribs (Individual) L3 1st L4 2nd L5 11th & 12th
254 Appendix C: Raw Data (Continued) Inventory of Individual Skeletons (Continued) Individual b, Se rra e Sa Caudeba Tomb B (Continued): Long Bones Proximal Epiphysis Proximal Third Middle Third Distal Third Distal Epiphysis Left Humerus 1 1 1 1 Right Humerus 1 1 1 1 Left Radius 1 1 1 1 Right Radius 1 1 1 1 Left Ulna 1 1 1 1 Right Ulna 1 1 1 1 Left Femur 1 1 1 1 1 Right Femur 1 1 1 1 1 Left Tibia 1 1 1 1 Right Tibia 1 1 1 1 Left Fibula 1 1 1 1 Right Fibula 1 1 1 1 Left Talus Right Talus 1 Left Calcaneus 1 Right Calcaneus Hand (# Present/# Complete) Foot (# Present/# Complete) L R U L R U #Carpals #Tarsals #Metacarpals #Metatarsals #Phalanges 11/0 #Phalanges = unsided ** = highly fragmented, could not identify to specific bone *** = highly fragmented, counted and grouped
255 Appendix C: Raw Data (Continued) Inventory of Individual Skeletons (Continued) Individual c, Serra e Sa Caudeba Tomb B: Cranial Elements Left Right Left Right Frontal Sphenoid Parietal Zygomatic Occipital Maxilla Temporal Palatine TMJ Mandible Postcranial Elements Left Right Left Right Clavicle 1 1 Os Coxae Scapula body 3 Auricular Surface 1 Glenoid f. Ilium 2 Patella 1 1 Ischium Sacrum 2* Pubis Acetabulum Vertebrae (Individual) Vertebrae (Grouped) # Present/ # Complete Centrum Neural Arch Centra Neural Arches C1 C3-6 2** C2 T1-T9 9** C7 Manubrium Body T10 Sternum T11 Ribs (Grouped) # Present/# Complete T12 L R U L1 3-10 52** L2 Ribs (Individual) L3 1st 1 1 L4 2nd L5 11th & 12th
256 Appendix C: Raw Data (Continued) Inventory of Individual Skeletons (Continued) Individual c, Serra e Sa Caudeba Tomb B (Continued): Long Bones Proximal Epiphysis Proximal Third Middle Third Distal Third Distal Epiphysis Left Humerus 1 1 1 1 1 Right Humerus 1 1 1 1 1 Left Radius 1 1 1 1 1 Right Radius 1 1 1 1 1 Left Ulna 1 1 1 1 1 Right Ulna 1 1 1 1 1 Left Femur 3 1 1 1 1 Right Femur 3 1 1 1 1 Left Tibia 1 1 1 1 Right Tibia 1 1 2 2 Left Fibula 1 1 1 1 Right Fibula 1 1 1 1 Left Talus Right Talus Left Calcaneus Right Calcaneus Hand (# Present/# Complete) Foot (# Present/# Complete) L R U L R U #Carpals #Tarsals #Metacarpals #Metatarsals #Phalanges 4/0 #Phalanges 5/0 = unsided ** = highly fragmented, could not identify to specific bone Comments: A bag with approximately 40 g of scarti was included with this individual. Also included were 35 small vertebral body fr agments that could not be identified to a more specific category.
257 Appendix C: Raw Data (Continued) Inventory of Individual Skeletons (Continued) Individual eI, Se rra e Sa Caudeba Tomb B: Cranial Elements Left Right Left Right Frontal Sphenoid Parietal Zygomatic Occipital Maxilla Temporal Palatine TMJ Mandible Postcranial Elements Left Right Left Right Clavicle Os Coxae Scapula body 2 Auricular Surface Glenoid f. 1 Ilium Patella Ischium Sacrum Pubis Acetabulum Vertebrae (Individual) Vertebrae (Grouped) # Present/ # Complete Centrum Neural Arch Centra Neural Arches C1 C3-6 C2 T1-T9 C7 Manubrium Body T10 Sternum T11 Ribs (Grouped) # Present/# Complete T12 L R U L1 3-10 L2 Ribs (Individual) L3 1st L4 2nd L5 11th & 12th
258 Appendix C: Raw Data (Continued) Inventory of Individual Skeletons (Continued) Individual eI, Se rra e Sa Caudeba Tomb B (Continued): Long Bones Proximal Epiphysis Proximal Third Middle Third Distal Third Distal Epiphysis Left Humerus Right Humerus Left Radius 1 1 1 Right Radius Left Ulna 1 Right Ulna 1 Left Femur 1 Right Femur 1 Left Tibia Right Tibia Left Fibula Right Fibula Left Talus Right Talus Left Calcaneus Right Calcaneus Hand (# Present/# Complete) Foot (# Present/# Complete) L R U L R U #Carpals #Tarsals #Metacarpals #Metatarsals #Phalanges #Phalanges
259 Appendix C: Raw Data (Continued) Inventory of Individual Skeletons (Continued) Individual eII, Se rra e Sa Caudeba Tomb B: Cranial Elements Left Right Left Right Frontal Sphenoid Parietal Zygomatic Occipital Maxilla Temporal Palatine TMJ Mandible Postcranial Elements Left Right Left Right Clavicle Os Coxae Scapula body 3* Auricular Surface Glenoid f. Ilium Patella Ischium Sacrum Pubis Acetabulum Vertebrae (Individual) Vertebrae (Grouped) # Present/ # Complete Centrum Neural Arch Centra Neural Arches C1 C3-6 C2 T1-T9 C7 Manubrium Body T10 Sternum T11 Ribs (Grouped) # Present/# Complete T12 L R U L1 3-10 L2 Ribs (Individual) L3 1st L4 2nd L5 11th & 12th
260 Appendix C: Raw Data (Continued) Inventory of Individual Skeletons (Continued) Individual eII, Se rra e Sa Caudeba Tomb B (Continued): Long Bones Proximal Epiphysis Proximal Third Middle Third Distal Third Distal Epiphysis Left Humerus Right Humerus Left Radius Right Radius Left Ulna 2* 2* Right Ulna Left Femur 1 1 1 2* Right Femur Left Tibia Right Tibia Left Fibula Right Fibula Left Talus Right Talus Left Calcaneus Right Calcaneus Hand (# Present/# Complete) Foot (# Present/# Complete) L R U L R U #Carpals #Tarsals #Metacarpals #Metatarsals #Phalanges #Phalanges = unsided Comments: An unidentifiable long-bone fragment was also included with this individual, but the size indicates that it does not belong to skeleton.
261 Appendix C: Raw Data (Continued) Inventory of Individual Skeletons (Continued) Individual eIII, Serra e Sa Caudeba Tomb B: Cranial Elements Left Right Left Right Frontal Sphenoid Parietal Zygomatic Occipital Maxilla Temporal Palatine TMJ Mandible Postcranial Elements Left Right Left Right Clavicle Os Coxae Scapula body Auricular Surface Glenoid f. Ilium Patella Ischium Sacrum Pubis Acetabulum Vertebrae (Individual) Vertebrae (Grouped) # Present/ # Complete Centrum Neural Arch Centra Neural Arches C1 C3-6 C2 T1-T9 C7 Manubrium Body T10 Sternum T11 Ribs (Grouped) # Present/# Complete T12 L R U L1 3-10 1* L2 Ribs (Individual) L3 1st L4 2nd L5 11th & 12th
262 Appendix C: Raw Data (Continued) Inventory of Individual Skeletons (Continued) Individual eIII, Serra e Sa Caudeba Tomb B (Continued): Long Bones Proximal Epiphysis Proximal Third Middle Third Distal Third Distal Epiphysis Left Humerus 2 Right Humerus 2 1 1 1 1 Left Radius 2* Right Radius Left Ulna Right Ulna Left Femur Right Femur Left Tibia 2* Right Tibia Left Fibula Right Fibula Left Talus 2* Right Talus Left Calcaneus Right Calcaneus Hand (# Present/# Complete) Foot (# Present/# Complete) L R U L R U #Carpals #Tarsals #Metacarpals #Metatarsals #Phalanges 3/1 #Phalanges = unsided
263 Appendix C: Raw Data (Continued) Inventory of Individual Skeletons (Continued) Individual , Se rra e Sa Caudeba Tomb B: Cranial Elements Left Right Left Right Frontal Sphenoid Parietal Zygomatic Occipital Maxilla Temporal Palatine TMJ Mandible Postcranial Elements Left Right Left Right Clavicle Os Coxae Scapula body 3 Auricular Surface Glenoid f. Ilium Patella 1 1 Ischium Sacrum 1 1 Pubis Acetabulum Vertebrae (Individual) Vertebrae (Grouped) # Present/ # Complete Centrum Neural Arch Centra Neural Arches C1 C3-6 C2 T1-T9 2/2 13* C7 Manubrium Body T10 Sternum T11 Ribs (Grouped) # Present/# Complete T12 L R U L1 3-10 20* L2 Ribs (Individual) L3 1st L4 2nd L5 11th & 12th
264 Appendix C: Raw Data (Continued) Inventory of Individual Skeletons (Continued) Individual , Se rra e Sa Caudeba Tomb B (Continued): Long Bones Proximal Epiphysis Proximal Third Middle Third Distal Third Distal Epiphysis Left Humerus 1 1 1 1 1 Right Humerus Left Radius 1 1 3 Right Radius 1 1 1 Left Ulna 2 3 Right Ulna 3 2 2 2 3 Left Femur 1 1 1 1 1 Right Femur 1 1 1 1 Left Tibia 1 1 1 1 1 Right Tibia 1 1 1 1 1 Left Fibula 1 1 1 1 Right Fibula 1 1 1 1 1 Left Talus 1 Right Talus Left Calcaneus 1 Right Calcaneus Hand (# Present/# Complete) Foot (# Present/# Complete) L R U L R U #Carpals #Tarsals #Metacarpals #Metatarsals #Phalanges 12/0 #Phalanges 4/0 = highly fragmented, could not identify to specific bone Comments: An additional 53 vertebral body fragments too small to identify and one lumbar body fragment that could not be clas sified further were included with this individual.
265 Appendix C: Raw Data (Continued) Table C.1. Pathologies Obser ved on Individual Skeletons Individual Bone or Element Conditions Observed Side Codes Photo # b Vertebral fragment Schmorl's node na 7.1.3 10 b Vertebral fragment Gradual change in body height, bone loss, osteophytes (barely discernable) Thoracic 1.8.2; 3.1.4; 3.2.1; 7.2.1 9 b Vertebral fragment Osteophytes na 7.2.2 11 b Vertebral fragment Schmorl's node na 7.1.2 N/A b Fibula Spicule/myositis ossificans Left 4.2.1; 4.6.1 7 b Rib Periostosis 4.2.1; 4.6.1 8 b Femur Myositis ossificans Left 4.2.1; 4.6.1 6 c Fibula Fracture Right 4.2.1; 4.6.1 47 c Humerus Arthritis Right 8.1.1; 8.2.2; 8.3.3 48, 49 c Vertebral fragment Osteophytes na 7.2.1 45 c Vertebral fragment Gradual change in body height; Osteophytes na 1.8.2; 7.2.1 44 c Vertebral fragment Osteophytes na 7.2.1 43 c Vertebral fragment Syndesmophytes na 7.3.3 42 c Vertebral fragment Osteophytes na 7.2.1 41 c Ilium Lytic lesions, multifocal Right 51, 52, 53 c Clavicle Fracture Left 46 eI Glenoid fossa Dislocation Right 8.5.3; 8.6.2 31, 32
266 Appendix C: Raw Data (Continued) Table C.1. Pathologies Observed on Individual Skeletons (Continued) Individual Bone or Element Conditions Observed Side Codes Photo # 6 Tibiae Reactive woven bone, porosity, eburnation bilateral, proximal 4.1.1; 8.4.2; 8.5.2; 8.6.1 167 6 Fibula Myositis ossificans U, proximal 5.4.9 168 6 Vertebral fragment Schmorl's node; osteophytes; spicule formation Thoracic 7.1.3; 8.1.2; 8.2.3 170 6 Vertebral fragment Schmorl's node Thoracic7.1.3 169 6 Humerus Osteoarthritis Right 8.5.3 171 6 Femur, head Osteoarthritis Left 8.5.3; 8.6.1 172 6 Femur, head Osteoarthritis Right 8.5.3; 8.6.1 173 Codes are after Buikstra and Ubelaker (1994: 114-115) and provide further description of the pathologies observed.
267 Appendix C: Raw Data (Continued) Table C.2. Raw Data from Cranial Elements Observation Number Context Bone Pathology Noted Side 1 Adolescenti and bambini Cranial fragment Porotic hyperostosis N/A 2 Adolescenti and bambini Occipital fragment Porotic hyperostosis 3 Cranium *unknown, possible child N/A No pathology observed N/A 4 Cranium? Frontal fragment Cribra orbitalia Left 5 Cranium 1 N/A No pathology observed N/A 6 Cranium 1? Temporal fragment Probable infection; mastoid process Left 7 Cranium 1? Temporal fragment Probable infection; mastoid process Right 8 Cranium 1? Cranial fragment Porotic hyperostosis 9 Cranium 1? Frontal fragment Porotic hyperostosis Right 10 Cranium 1? Frontal fragment Cribra orbitalia Right 11 Cranium 10 N/A No pathology observed N/A 12 Cranium 10? N/A No pathology observed N/A 13 Cranium 11 N/A No pathology observed N/A 14 Cranium 11? Cranial fragment Porotic hyperostosis 15 Cranium 12 Parietal fragment Porotic hyperostosis 16 Cranium 12? N/A No pathology observed N/A 17 Cranium 13 Cranial fragments (3) Porotic hyperostosis N/A 18 Cranium 13? N/A No pathology observed N/A 19 Cranium 14 N/A No pathology observed N/A
268 Appendix C: Raw Data (Continued) Table C.2. Raw Data from Cranial Elements (Continued) Observation Number Context Bone Pathology Noted Side 20 Cranium 14 Cranial fragment Porotic hyperostosis U 21 Cranium 15 Cranial fragment Porotic hyperostosis U 22 Cranium 15? Frontal fragment Cribra orbitalia Right? 23 Cranium 16 N/A No pathology observed N/A 24 Cranium 16? N/A No pathology observed N/A 25 Cranium 17 N/A No pathology observed N/A 26 Cranium 17? N/A No pathology observed N/A 27 Cranium 19 Occipital fragment Porotic hyperostosis U 28 Cranium 2 Cranial fragment Porotic hyperostosis U 29 Cranium 2 (l2-e2?) Frontal fragment Cribra orbitalia 30 Cranium 2 (l2-e2?) Occipital fragment Porotic hyperostosis 31 Cranium 2 (l2-e2?) Parietal fragments (10) Porotic hyperostosis 32 Cranium 20 Cranial fragment Lesion, endocranial N/A 33 Cranium 20 Cranial fragment Porotic hyperostosis N/A 34 Cranium 21 N/A No pathology observed N/A 35 Cranium 22 N/A No pathology observed N/A 36 Cranium 23 Cranial fragment Porotic hyperostosis U 37 Cranium 24 N/A No pathology observed N/A 38 Cranium 25 N/A No pathology observed N/A
269 Appendix C: Raw Data (Continued) Table C.2. Raw Data from Cranial Elements (Continued) Observation Number Context Bone Pathology Noted Side 39 Cranium 26 N/A No pathology observed N/A 40 Cranium 27 N/A No pathology observed N/A 41 Cranium 28 N/A No pathology observed N/A 42 Cranium 29 N/A No pathology observed N/A 43 Cranium 3 Cranial fragment Porotic hyperostosis U 44 Cranium 3 Frontal fragmentCribra orbitalia Right 45 Cranium 3?? Cranial fragment Porotic hyperostosis 46 Cranium 3? Frontal fragmentCribra orbitalia Left 47 Cranium 3? Frontal fragmentPorotic hyperostosis 48 Cranium 3? Parietal fragment Porotic hyperostosis 49 Cranium 30 N/A No pathology observed N/A 50 Cranium 31 N/A No pathology observed N/A 51 Cranium 32 N/A No pathology observed N/A 52 Cranium 33 N/A No pathology observed N/A 53 Cranium 34 N/A No pathology observed N/A 54 Cranium 35 N/A No pathology observed N/A 55 Cranium 36 Cranial fragment Porotic hyperostosis U 56 Cranium 37 N/A No pathology observed N/A 57 Cranium 38 N/A No pathology observed N/A 58 Cranium 4? N/A No pathology observed N/A 59 Cranium 40 N/A No pathology observed N/A
270 Appendix C: Raw Data (Continued) Table C.2. Raw Data from Cranial Elements (Continued) Observation Number Context Bone Pathology Noted Side 60 Cranium 41 N/A No pathology observed N/A 61 Cranium 42 N/A No pathology observed N/A 62 Cranium 43? Cranial fragment Porotic hyperostosis 63 Cranium 44 N/A No pathology observed N/A 64 Cranium 45 N/A No pathology observed N/A 65 Cranium 45 N/A No pathology observed N/A 66 Cranium 46 N/A No pathology observed N/A 67 Cranium 5? N/A No pathology observed N/A 68 Cranium 6 N/A No pathology observed N/A 69 Cranium 6? N/A No pathology observed N/A 70 Cranium 7 N/A No pathology observed N/A 71 Cranium 7? N/A No pathology observed N/A 72 Cranium 8 Cranial fragment Porotic hyperostosis 73 Cranium 8 Frontal fragmentCribra orbitalia Left 74 Cranium 9 Cranial fragment Porotic hyperostosis 75 Cranium a1 N/A No pathology observed N/A 76 Cranium c1 N/A No pathology observed N/A 77 Cranium C2? Frontal fragme ntCribra orbitalia Right 78 Cranium C2? Parietal fragment Porotic hyperostosis Right 79 Cranium c-e N/A No pathology observed N/A
271 Appendix C: Raw Data (Continued) Table C.2. Raw Data from Cranial Elements (Continued) Observation Number Context Bone Pathology Noted Side 80 Cranium d Cranial fragment Porotic hyperostosis N/A 81 Cranium d1 Cranial fragment Porotic hyperostosis 82 Cranium g N/A No pathology observed N/A 83 Cranium g1 N/A No pathology observed N/A 84 Cranium ie1 Frontal fragmentPorotic hyperostosis 85 Cranium ie1 Frontal fragmentCribra orbitalia 86 Cranium l1 N/A No pathology observed N/A 87 Cranium m1 N/A No pathology observed N/A 88 Cranium M1 Cranial fragment Porotic hyperostosis 89 Cranium O1 N/A No pathology observed N/A 90 Cranium p1 N/A No pathology observed N/A 91 Cranium q Occipital fragment Porotic hyperostosis 92 Cranium r1 N/A No pathology observed N/A 93 Cranium s N/A No pathology observed N/A 94 Cranium t N/A No pathology observed N/A 95 Cranium t? Cranial fragment Porotic hyperostosis 96 Cranium t1 N/A No pathology observed N/A 97 Cranium t2 Cranial fragment Porotic hyperostosis 98 Cranium v N/A No pathology observed N/A 99 Cranium Z Cranial fragment Porotic hyperostosis
272 Appendix C: Raw Data (Continued) Table C.2. Raw Data from Cranial Elements (Continued) Observation Number Context Bone Pathology Noted Side 100 No Number A Cranial fragment Porotic hyperostosis 101 No Number B N/A No pathology observed N/A 102 No Number C N/A No pathology observed N/A 103 No Number D Cranial fragment Porotic hyperostosis 104 No Number E Cranial fragment Porotic hyperostosis 105 No Number E Frontal fragment Cribra orbitalia 106 No Number F Frontal fragment Cribra orbitalia 107 No Number G Cranial fragment Porotic hyperostosis 108 No Number H N/A No pathology observed N/A 109 No Number I Frontal fragment Cribra orbitalia 110 No Number I Temporal fragment Porotic hyperostosis 111 No Number I Occipital fragment Porotic hyperostosis 112 No Number J N/A No pathology observed N/A 113 No Number K Parietal fragment Porotic hyperostosis 114 No Number L N/A No pathology observed N/A 115 Paretes (?) del taplio stradale Cranial fragment Porotic hyperostosis 116 Paretes (?) del taplio stradale Frontal fragment Cribra orbitalia Left 117 Q11 (-120-130) Cranial fragment Porotic hyperostosis 118 Q11 (-160) Cranial fragment Porotic hyperostosis
273 Appendix C: Raw Data (Continued) Table C.2. Raw Data from Cranial Elements (Continued) Observation Number Context Bone Pathology Noted Side 119 Q11 (-160) Mandibular fragment Carnivore activity Right 120 Q11 (-160) Cranium 13 Cranial fragment Porotic hyperostosis 121 Q11 (-160) Cranium 13 Cranial fragment Cribra orbitalia 119 Q11 (-160) Mandibular fragment Carnivore activity Right 120 Q11 (-160) Cranium 13 Cranial fragment Porotic hyperostosis 121 Q11 (-160) Cranium 13 Cranial fragment Cribra orbitalia 122 Q11 (-160) Cranium 15 Occipital fragment Porotic hyperostosis 123 Q11 (-160) Cranium 15 Parietal fragment Porotic hyperostosis 124 Q12 Q14 Cranial fragment Porotic hyperostosis 125 Q12 Q14 Frontal fragment Cribra orbitalia 126 Q12 (-160) Occipital No pathology observed 127 Q12 (-160) Cranial fragment Porotic hyperostosis 128 Q12 (-160) Parietal fragments (4) Porotic hyperostosis 129 Q12 (-160) Occipital Porotic hyperostosis 130 Q13 (-140) Cranial fragments (4) Porotic hyperostosis 131 Q13 (-140) Parietal Porotic hyperostosis Right 132 Q13 (-150) Cranial fragment Porotic hyperostosis 133 Q15 Frontal fragment Porotic hyperostosis 134 Q15 (-130) Cranial fragment Porotic hyperostosis 135 Q16 (-130) Parietal fragment Porotic hyperostosis
274 Appendix C: Raw Data (Continued) Table C.2. Raw Data from Cranial Elements (Continued) Observation Number Context Bone Pathology Noted Side 136 Q17 (-130) Cranial fragment Porotic hyperostosis 137 Q18 (-130) Cranial fragment Porotic hyperostosis 138 Q2 Q4 (-155 to 170) Cranial fragment Porotic hyperostosis N/A 139 Q5 zona est (-155 165) Cranial fragment Porotic hyperostosis 136 Q17 (-130) Cranial fragment Porotic hyperostosis 137 Q18 (-130) Cranial fragment Porotic hyperostosis 138 Q2 Q4 (-155 to 170) Cranial fragment Porotic hyperostosis N/A 139 Q5 zona est (-155 165) Cranial fragment Porotic hyperostosis 140 Q6 (-180) Mandibular fragment No pathology observed Right 141 Q6 (-180) Temporal fragment No pathology observed Right 142 Q6 (-180) Cranium 3 Cranial fragment Porotic hyperostosis 143 Q6 (-180) Cranium 5 N/A No pathology observed N/A 144 Q9 (-180) Frontal fragment Cribra orbitalia Left 145 Sepolture di fronte alle tombe B (cranio) Cranial fragments No pathology observed N/A 146 Taglio de -130 to 140 sul lato dentro della gallerie Cranial fragment Lesion, endocranial 147 Taglio de -130 to 140 sul lato dentro della gallerie Frontal fragment Cribra orbitalia Right 148 taglio longitudinale al liello del parsmento? Lato scarpato? Cranial fragment Porotic hyperostosis
275 Appendix C: Raw Data (Continued) Table C.2. Raw Data from Cranial Elements (Continued) Observation Number Context Bone Pathology Noted Side 149 Taglio nella terra piella sup. Cranial fragment/occi pital bone Healed depressed fracture 150 W2 W3 (a ebuteru o col povronemio lotu seorpere?) Cranial fragment Lesion, endocranial 151 W2 W3 (a ebuteru o col povronemio lotu seorpere?) Cranial fragment Porotic hyperostosis 152 X1 X2 X3 Cranial fragment Porotic hyperostosis
276 Appendix C: Raw Data (Continued) Table C.3. Raw Data from Individual Elements Observation Number Context Bone Category 1 W4 X4 (primo toglio) Long bone fragment Abnormal Bone Formation 2 Z2 (primo toglio) Long bone fragment Abnormal Bone Formation 3 Q16 (-130) Long bone fragment Abnormal Bone Formation 4 Femora Femur Abnormal Bone Formation 5 Q3 (-180) Fibula Abnormal Bone Formation 6 No context Ulna Abnormal Bone Formation 7 Q3 (-180) Long bone fragment Abnormal Bone Formation 8 Q10 (-180) Sternum Abnormal Bone Loss 9 Q13 (-130) Ulna Abnormal Bone Loss 10 Q0 (-185) Calcaneus Abnormal Bone Loss 11 Q1 (-180 240) Cervical vertebra Abnormal Bone Loss 12 Q1 (-180) Sternum (manubrium) Abnormal Bone Loss 13 Q1 (-180 240) Vertebral fragment Abnormal Bone Loss 14 Q3 (-180) Vertebral fragment Abnormal Bone Loss 15 Q3 (-180) Vertebral fragment (thoracic) Abnormal Bone Loss 16 Q4 (-180) Vertebral fragment Abnormal Bone Loss 17 Q4 (-180) Vertebral fragment Abnormal Bone Loss 18 Q5 (-155-165) Vertebral fragment Abnormal Bone Loss 19 Vertebra Vertebral fragment Abnormal Bone Loss
277 Appendix C: Raw Data (Continued) Table C.3. Raw Data from Individual Elements (Continued) Observation Number Context Bone Category 20 W2 W3 (a ebuteru o col povronemio lotu seorpere?) Vertebral fragment Abnormal Bone Loss 21 W2 W3 (a ebuteru o col povronemio lotu seorpere?) Vertebral fragment Abnormal Bone Loss 22 W2 W3 (a ebuteru o col povronemio lotu seorpere?) Vertebral fragment Abnormal Bone Loss 23 W2 W3 (a ebuteru o col povronemio lotu seorpere?) Vertebral fragment Abnormal Bone Loss 24 Q1 (-180) Vertebral fragment (thoracic) Abnormal Bone Loss 25 Q5 (-175) Vertebral fragment (pedicle) Abnormal Bone Loss 26 Q13 (-130) Vertebral fragment (centra) Abnormal Bone Loss 27 Q12 (-160) Cervical vertebrae Abnormal Bone Loss 28 Q12 (-160) Cervical vertebrae Abnormal Bone Loss 29 Q9 (-170) Navicular Abnormal Bone Loss 30 Q6 (-180) Vertebral fragment (thoracic) Abnormal Bone Loss 31 Q3 (-180) Long bone fragment Abnormal Bone Loss 32 Q13 (-140) Femur Abnormal Bone Loss 33 Q12 (-140) Humerus Abnormal Bone Loss (possible) 34 Q12 (-180) Humerus Abnormal Bone Loss (possible) 35 1st Scatola Vertebral fragment Abnormality of Shape 36 1st Scatola Vertebrae Abnormality of Shape
278 Appendix C: Raw Data (Continued) Table C.3. Raw Data from Individual Elements (Continued) Observation Number Context Bone Category 37 Femora Femur Arthritis 38 Illium Acetabulum Arthritis 39 Q13 (-140) Humerus Arthritis 40 Q12 Q14 Phalanx Arthritis 41 Ulnae Ulna Arthritis 42 Q5 (-180) Vertebral fragment (thoracic) Arthritis 43 Q17 (-130) Femur (head) Arthritis 44 Femora Femur Arthritis 45 Femora Femur Arthritis 46 Femora Femur Arthritis 47 Femora Femur Arthritis 48 Q15 (-130) Ulna Arthritis 49 taglio longitudinale al liello del parsmento? Lato scarpato? Femur, head Arthritis 50 taglio longitudinale al liello del parsmento? Lato scarpato? Femur, head (?) Arthritis 51 Adolescenti and bambini Humerus? Fractures and Dislocations 52 Q13 (-140) Femoral head Multiple 53 Q13 (-140) Humerus Multiple 54 Q11 (-120-130) Long bone fragmentMultiple 55 1st Scatola Vertebral fragment Multiple 56 Q5 (-175) Vertebral fragment Multiple 57 1st Scatola Vertebrae Multiple 58 Q4 (-180) Vertebral fragment, Multiple 59 Q4 (-180) Vertebral fragment Taphonomy 60 Q17 (-130) Long bone fragmentTaphonomy 61 Tibiae Long bone fragmentTaphonomy 62 W2 W3 (a ebuteru o col povronemio lotu seorpere?) Long bone fragmentTaphonomy 63 Radius Radial head Taphonomy 64 Q0 (Ingresso 195) Vertebral fragment Vertebral Pathology
279 Appendix C: Raw Data (Continued) Table C.3. Raw Data from Individual Elements (Continued) Observation Number Context Bone Category 65 Q12 Q14 Vertebral fragmentVertebral Pathology 66 Q16 (-130) Vertebral fragmentVertebral Pathology 67 1st Scatola Vertebral fragmentVertebral Pathology
280 Appendix C: Raw Data (Continued) Table C.3. Raw Data from Individual Elements (Continued) Observation Number Categories (multiple) Pathologies Noted Side Codes (from Buikstra and Ubelaker 1994:114115) 1 Hyperostosis 4.5.2; 4.6.3 2 Hyperostosis 4.5.2; 4.6.3 3 Hyperostosis 4.2.2; 4.5.2; 4.6.1 4 Periostitis Left 8.7.2 5 Cloaca, myositis ossificans? 3.1.4; 3.2.1; 3.3.1; 3.4.1; 3.5.2 6 Cloaca Left 3.1.4; 3.2.1; 3.3.1; 3.4.1; 3.5.1 7 Treponematosis? 5.1.4; 5.4.2 8 Lesions 3.1.4; 3.2.1; 3.3.2; 3.4.4; 3.5.4 9 Lytic lesion/hyperostosis? Left 3.1.4; 3.2.1; 3.3.1; 3.4.2; 3.5.6 10 Lytic lesion Left 3.1.4; 3.2.2; 3.3.3; 3.4.7; 3.5.6 11 Lytic lesion 3.1.2; 3.2.1; 3.3.1; 3.4.1 12 Lytic lesion 3.1.4; 3.2.1; 3.3.1; 3.4.2; 3.5.2 13 Lytic lesion top of body (rephrase) 3.1.2; 3.2.1; 3.3.1; 3.4.1 14 Lytic lesion 3.1.2; 3.2.1; 3.3.2; 3.4.2; 3.5.3 15 Lytic lesion 3.1.2; 3.2.1; 3.3.2; 3.4.1; 3.5.2
281 Appendix C: Raw Data (Continued) Table C.3. Raw Data from Individual Elements (Continued) Observation Number Categories (multiple) Pathologies Noted Side Codes (from Buikstra and Ubelaker 1994:114115) 16 Lytic lesion 3.1.2; 3.2.1; 3.3.1; 3.4.1; 3.5.2 17 Lytic lesion 3.1.4; 3.2.1; 3.3.3; 3.4.1; 3.5.2 18 Lytic lesion 3.1.2; 3.2.1; 3.3.1; 3.4.1; 3.5.3 19 Lytic Lesion N/A 3.1.4; 3.2.2; 3.3.2; 3.4.2; 3.5.6 20 Lytic lesion 21 Lytic lesion 3.1.4; 3.2.1; 3.3.1; 3.4.1; 3.5.2 22 Lytic lesion 3.1.4; 3.2.1; 3.3.1; 3.4.0; 3.5.3 23 Lytic lesion 3.1.4; 3.2.1; 3.3.1; 3.4.1; 3.5.3 24 Lytic lesion 3.1.2; 3.2.1; 3.3.1; 3.4.1; 3.5.3 25 Lytic lesion 3.1.4; 3.2.1; 3.3.1; 3.4.1; 3.5.3 26 Lytic lesion 3.1.4; 3.2.1; 3.3.1; 3.4.2; 3.5.3 27 Porosity 3.6.2 28 Porosity 3.6.2
282 Appendix C: Raw Data (Continued) Table C.3. Raw Data from Individual Elements (Continued) Observation Number Categories (multiple) Pathologies Noted Side Codes (from Buikstra and Ubelaker 1994:114115) 29 Possible lesions Left 3.1.4; 3.2.1; 3.3.1; 3.4.2; 3.5.3 30 Lytic lesion 3.1.2; 3.2.1; 3.3.2; 3.4.1; 3.5.3 31 Treponematosis? 5.1.4; 5.4.2 32 Necrosis? Right 3.1.2; 3.2.1; 3.3.1; 3.4.1; 3.5.2; 4.2.1; 4.6.1 33 Lytic lesion Right 3.1.4; 3.2.2; 3.3.1; 3.4.2; 3.5.3 34 Lytic lesion 3.1.2; 3.2.1; 3.3.1; 3.4.1; 3.5.3 35 Slight angling of body 1.8.2 36 Ankylosis Axis and 3rd CV 1.7.1; 1.8.1; 1.9.1 37 Arthritis Left 8.5.3; 8.6.3 38 Lipping Right 8.1.0; 8.4.1 39 Lipping Left 8.1.2; 8.4.1 40 Lipping 8.1.3; 8.2.2; 8.3.3 41 Lipping 8.1.3; 8.2.1 42 Lipping 8.1.2; 8.2.2 43 Lipping and bony response 8.1.2; 8.2.3; 8.4.1 44 Lipping at distal end U 8.1.2; 8.2.2 45 Lipping at distal end U 8.1.2; 8.2.2
283 Appendix C: Raw Data (Continued) Table C.3. Raw Data from Individual Elements (Continued) Observation Number Categories (multiple) Pathologies Noted Side Codes (from Buikstra and Ubelaker 1994:114115) 46 Lipping at distal end U 8.1.2; 8.2.2 47 Lipping at distal end U 8.1.2; 8.3.3 48 Arthritis 5.1.2; 5.4.2; 5.4.7 49 Arthritis U 3.1.4; 3.2.2; 3.3.1; 3.4.2; 3.5.7; 8.3.2; 8.5.3 50 Arthritis U 3.1.4; 3.2.2; 3.3.1; 3.4.2; 3.5.7; 8.3.2; 8.5.3 51 Fracture (distal) 5.1.2; 5.4.2 52 Abnormal Bone Loss/Arthritis Lesion, arthritis? U 3.1.4; 3.2.1; 3.3.3; 3.5.5 53 Abnormal Bone Loss/Arthritis Lesion, arthritis Left 3.1.4; 3.2.1; 3.3.1; 3.4.2; 3.5.3 54 Abnormal Bone Shape, Taphonomy hyperostosis, excavation damage? 1.3.2; 1.4.2 55 Abnormal Bone Loss, Fractures and Dislocations Lytic lesions with wedge-shaped angling of vertebra, compression fx? ? 1.8.2; 3.1.2; 3.2.2; 3.3.1; 3.4.2; 3.5.3 56 Abnormal Bone Loss, Vertebral Pathology Lytic lesion Lumbar 3.1.4; 3.2.1; 3.3.1; 3.4.2; 3.5.2; 7.1.3;7.2.3
284 Appendix C: Raw Data (Continued) Table C.3. Raw Data from Individual Elements (Continued) Observation Number Categories (multiple) Pathologies Noted Side Codes (from Buikstra and Ubelaker 1994:114115) 57 Abnormal Bone Loss, Fractures and Dislocations Lytic lesions with wedge-shaped angling of vertebra, compression fx? Thoracic/Lumbar ? 1.7.1; 1.8.1; 1.9.1; 3.1.4; 3.2.1; 3.3.1; 3.4.1; 3.5.1 58 Abnormal Bone Loss, Fractures and Dislocations Lytic lesion with wedge-shaped angling of vertebra, compression fx? Thoracic 3.1.2; 3.2.1; 3.3.1; 3.4.2; 3.5.3;1.8.1; 1.9.1 59 Insect activity 3.1.4; 3.2.1; 3.3.1; 3.4.1; 3.5.2 60 Rodent gnawing 61 Rodent gnawing 62 Rodent gnawing 63 Carnivore activity 64 Lipping 7.2.2 65 Schmorl's node 7.1.3 66 Schmorl's node 7.1.2 67 Syndesmophytes ? 7.3.1
285 Appendix D: Photographs of Pathologies Observed on the Individual Skeletons Pathologies observed on elements fr om the individual skeletons were photographed and are included in this appe ndix. The photograph numbers are from the data presented in Appendix C, Pathologies Observed on Individual Skeletons. In the event of multiple photographs of an element, representative images were selected. All photographs in this section were taken by the author.
286 Appendix D: Photographs of Pathologies Observed on the Individual Skeletons (Continued) Figure D.1. Photograph 6: myositis ossific ans of the left femur of individual b Figure D.2. Photograph 7: spicule/myositis os sificans of the left fi bula of individual b
287 Appendix D: Photographs of Pathologies Observed on the Individual Skeletons (Continued) Figure D.3. Photograph 8: periostosi s of the rib of individual b Figure D.4. Photograph 9: gradual change in bo dy height, bone loss, and osteophytes (barely discernable) of a thorac ic vertebrae of individual b
288 Appendix D: Photographs of Pathologies Observed on the Individual Skeletons (Continued) Figure D.5. Photograph 10: Schmorl's node of a vertebral fragment of individual b Figure D.6. Photograph 11: osteophytes of a vertebral fragment of individual b
289 Appendix D: Photographs of Pathologies Observed on the Individual Skeletons (Continued) Figure D.7. Photograph 42: syndesmophytes of a vertebral body of individual c Figure D.8. Photograph 43: osteophytes of a vertebral body of individual c
290 Appendix D: Photographs of Pathologies Observed on the Individual Skeletons (Continued) Figure D.9. Photograph 44: gradual change in body height and osteophytes of a vertebral body of individual c Figure D.10. Photograph 45: osteophytes of a vertebral body fragment of individual c
291 Appendix D: Photographs of Pathologies Observed on the Individual Skeletons (Continued) Figure D.11. Photograph 46:. fracture of the left clavicle of individual c Figure D.12. Photograph 47: fracture of the right fibula of individual c
292 Appendix D: Photographs of Pathologies Observed on the Individual Skeletons (Continued) Figure D.13. Photograph 48: arthritis of the right humerus of individual c Figure D.14. Photograph 49: arthritis of the right humerus of individual c
293 Appendix D: Photographs of Pathologies Observed on the Individual Skeletons (Continued) Figure D.15. Photograph 51: multifocal lytic lesi ons of the right il ium of individual c
294 Appendix D: Photographs of Pathologies Observed on the Individual Skeletons (Continued) Figure D.16. Photograph 32: dislocation of th e right glenoid fossa of individual eI Figure D.17. Photograph 167: reactive woven bone, porosity, and eburnation of the proximal tibiae of i ndividual 6 (bilateral)
295 Appendix D: Photographs of Pathologies Observed on the Individual Skeletons (Continued) Figure D.18. Photograph 168: myositis ossificans of a fibula of individual 6 Figure D.19. Photograph 169: Schmorl's node of a thoracic vertebrae of individual 6
296 Appendix D: Photographs of Pathologies Observed on the Individual Skeletons (Continued) Figure D.20. Photograph 170: Schmorl's node, os teophytes, and spicule formation were observed on a thoracic vertebra of individual 6 Figure D.21. Photograph 171:. osteoarthritis of the right humerus of individual 6
297 Appendix D: Photographs of Pathologies Observed on the Individual Skeletons (Continued) Figure D.22. Photograph 172: osteoarthritis of the left femoral head of individual 6 Figure D.23. Photograph 173: osteoarthritis of the left femoral head of individual 6
298 Appendix E: Photographs of Examples of Common Pathologies in this Population In addition to examining the indivi dual skeletons, cranial fragments and unassociated skeletal elements were examined. The following photographs are representative of the most common pathologies observed. All av ailable contextual information is provided (e.g., cranium number or depth of find); however, as discussed in Chapter Five, some bones lacked contextual information. No catalog numbers have been assigned to these remains. All photographs in this section were taken by the author.
299 Appendix E: Photographs of Examples of Common Pathologies in this Population (Continued) Figure E.1. Endocranial lesion (c ranial fragment from W2-W3) Figure E.2. Porotic hyperostosis (parietal from cranium e2)
300 Appendix E: Photographs of Examples of Common Pathologies in this Population (Continued) Figure E.3. Cribra orbitalia (cranium c2) Figure E.4. Angling of vertebral body (q4-180)
301 Appendix E: Photographs of Examples of Common Pathologies in this Population (Continued) Figure E.5. Lytic lesion of vertebral body (q4-180) Figure E.6. Arthritis of femur
302 Appendix F: Photographs of Examples of Unusual Pathologies in this Population Unique pathologies and those with fe wer occurrences were categorized as unusual. The following are photog raphs of unusual pathologies. All available contextual information is provided (e.g., cranium number or depth of find); however, as discussed in Chapter Five, some bones lacked contextual information. No catalog numbers have been assigned to these remains. All photographs in this section were taken by the author.
303 Appendix F: Photographs of Examples of Unusual Pathologies in this Population (Continued) Figure F.1. Hypoplasia of mastoi d process (presented as bila teral in mp1; right side pictured) Figure F.2. Meningeal reaction with granular impressions (cranium 3)
304 Appendix F: Photographs of Examples of Unusual Pathologies in this Population (Continued) Figure F.3. Depression fractu re of parietal bone (cranium taglio nella terra ) Figure F.4. Cloaca (tibia)
305 Appendix F: Photographs of Examples of Unusual Pathologies in this Population (Continued) Figure F.5. Fracture (juvenile humerus) Figure F.6. Lesion sternum (q1-180)
306 Appendix F: Photographs of Examples of Unusual Pathologies in this Population (Continued) Figure F.7. Necrosis of the femur (q13-140) A Figure F.8. Ankylosis of cervical vertebrae ( 1 scatola )
307 Appendix F: Photographs of Examples of Unusual Pathologies in this Population (Continued) Figure F.9. Lesions of the sternum Figure F.10. Arthritis (acetabulum)
308 Appendix F: Photographs of Examples of Unusual Pathologies in this Population (Continued) Figure F.11. Arthritis phalange (q12-q14 -140-160) Figure F.12. Arthritis (ulna)
309 Appendix F: Photographs of Examples of Unusual Pathologies in this Population (Continued) Figure F.13. Lytic lesi on navicular (q9) Figure F.14. Arthritis ulna (q15-130)
310 Appendix F: Photographs of Examples of Unusual Pathologies in this Population (Continued) Figure F.15. Lesion calcaneus (q0-185) Figure F.16. Lesion ulna (q13-140)
311 Appendix F: Photographs of Examples of Unusual Pathologies in this Population (Continued) Figure F.17. Myositis ossificans of an ul na, possibly from a fracture (fragment q3-180) Figure F.18. Associated bony reaction in the adjacent radius (fragment q3-180)
312 Appendix F: Photographs of Examples of Unusual Pathologies in this Population (Continued) Figure F.19. Porosity of vertebra ( 1 scatola) Figure F.20. Porosity of vertebra ( 1 scatola)
313 Appendix F: Photographs of Examples of Unusual Pathologies in this Population (Continued) Figure F.21. Hyperostosis of long bone fragment (w4-x4)
314 Appendix G: Photographs of Bones Used for Sampling for Ancient DNA, Pf HRP II, and Hemozoin Analysis Ten bone fragments were selected for aDNA, protein, and hemozoin testing. Detailed information about the selection and preparation of these samples is provided in Chapter Five. All available contextual info rmation is provided (e.g., cranium number or depth of find); however, as discussed pr eviously, some bones lacked contextual information. No catalog numbers have been as signed to these remains. All photographs in this section were taken by the author.
315 Appendix G: Photographs of Bones Used for Sampling for Ancient DNA, Pf HRP II, and Hemozoin Analysis (Continued) Figure G.1. Sample 1: right humerus fragment, infant ( bambini e adolescenti ) Figure G.2. Sample 2: left humerus, infant ( bambini e adolescenti )
316 Appendix G: Photographs of Bones Used for Sampling for Ancient DNA, Pf HRP II, and Hemozoin Analysis (Continued) Figure G.3. Sample 3: long bone fragment, infant ( bambini e adolescenti ) Figure G.4. Sample 4. Long bone fragment, infant ( bambini e adolescenti )
317 Appendix G: Photographs of Bones Used for Sampling for Ancient DNA, Pf HRP II, and Hemozoin Analysis (Continued) Figure G.5. Sample 5: left ulna fragment, adolescent ( sepoltura de fronte alla tomba B ) Figure G.6. Sample 6: left humer us fragment, adolescent (Q1; quota 180/240 tomba B )
318 Appendix G: Photographs of Bones Used for Sampling for Ancient DNA, Pf HRP II, and Hemozoin Analysis (Continued) Figure G.7. Sample 7: unsided humerus fragment, adolescent (Q16; quota 130 tomba B ) Figure G.8. Sample 8: unsided humerus fragment, adult (Q1; quota 180 tomba B )
319 Appendix G: Photographs of Bones Used for Sampling for Ancient DNA, Pf HRP II, and Hemozoin Analysis (Continued) Figure G.9. Sample 9: right humerus, adult (W4 ; ex primo taglio tomba B ) Figure G.10. Sample 10: right humerus, adult ( Quota 177 tomba B )
About the Author Teddi Setzer graduated with a bachel ors degree from the Department of Psychology at the University of South Flor ida in 1996. In 2001, she en tered the masters program in the Department of Anthropology at the University of South Florida, where she began studying the prehistory of the cen tral Mediterranean. Her thesis research involved studying use wear on Neolithic obs idian tools from Sardinia. After she graduated with her masters degree in 2004, sh e continued her doctoral studies in applied anthropology at the University of South Florida. Continued resear ch in archaeology and biological anthropology, as well as participa ting in fieldwork in Sardinia, piqued her interest in the health and lifeways of prehistoric Sardinian pop ulations. Her work involves examining the coevolution of the host, vector, and parasite in malaria infections and applying anthropological perspectives when identifying and examining biocultural adaptations to this disease.