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Title:
Before the Inca prehistoric dietary transitions in the Argentine Cuyo
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English
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Shelnut, Nicole
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University of South Florida
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Subjects / Keywords:
Archaeology
Archaeometry
Bone chemistry
Diet
Environment
Human ecology
Mendoza
Mummification
Nutrition
San Juan
South America
Stable isotope analysis
Dissertations, Academic -- Applied Anthropology -- Masters -- USF
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bibliography   ( marcgt )
theses   ( marcgt )
non-fiction   ( marcgt )

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Abstract:
ABSTRACT: A dietary reconstruction was performed in order to understand changing prehistoric subsistence patterns in the Central Andean geographical area of the Argentine Cuyo that includes the provinces of San Juan and Mendoza. Archaeologically, the Cuyo is also known as a boundary between Andean agriculturalists and the foragers of Patagonia. One hypothesis being tested is whether this area was one of the last South American cultural groups to convert to maize cultivation, probably around 2000 BP. The process of stable isotope analysis is used to reconstruct the diets of individuals, as it reveals the relative proportions of C3 and C4 plants and the contribution of aquatic resources to otherwise terrestrial diets, as well as variations in trophic level of the foods consumed.In this study the bones, teeth, hair, and flesh from 45 individuals were tested to address specifically total and protein diets, as well as seasonal variation and changes between childhood and adulthood. This process, when used in combination with previous analyses, such as midden or faunal analysis, allows researchers to evaluate the results of those previous studies, and thus compose a more thorough reconstruction of the lifestyles of a prehistoric culture.Information garnered from this study indicates that the times of dietary transition were variable, with seasonal patterns becoming more stable over long periods. Furthermore, some members of the study population demonstrate the existence of nutritional stress indicators, such as dental caries, that can be viewed in relation to the dietary shifts that may have been a cultural adaptation to the environment of the Cuyo. Overall, this study shows the early adoption of maize agriculture in central western Argentina and recommends future studies that analyze the relationships between agriculture, diet, and nutrition in the New World.
Thesis:
Thesis (M.A.)--University of South Florida, 2006.
Bibliography:
Includes bibliographical references.
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by Nicole Shelnut.
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Title from PDF of title page.
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Document formatted into pages; contains 126 pages.

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Before the Inca: Prehistoric Dietary Transitions in the Argentine Cuyo by Nicole Shelnut A thesis submitted in partial fulfillment of the requirements for the degree of Master of Arts Department of Anthropology College of Arts and Sciences University of South Florida Major Professor: Robert H. Tykot, Ph.D. David Himmelgreen, Ph.D. E. Christian Wells, Ph.D. Date of Approval: April 14, 2006 Keywords: archaeology, archaeometry, bone chemistry, diet, environment, human ecology, Mendoza, mummification, nutrition, Sa n Juan, South America, stable isotope analysis Copyright 2006, Nicole Shelnut

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For Roy E. Shelnut My father, my inspiration, my friend

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Acknowledgments Many thanks are given to the committee members who have worked so diligently and selflessly to assist in the realization of this work, in part icular Dr. Robert H. Tykot, as well as Dr. E. Christian Wells and Dr. Davi d Himmelgreen. This work could not have been accomplished without the aid of Drs. Adolfo Gil and Gustavo Neme, who have proven themselves as both ultimate colleagues and friends. They began and organized this collaborative study; their invariable, magnanimous help and willingness to share information made this work possible. Of course, constant encouragement was given by family members and friends including Coll een Regan, Florence Tosh, Corrine LeClaire, Teddi Setzer, and Elena Vaquera. Access to materials was provided by the Museo de Historia Natural, San Rafael, Argentina. Th is research was supported by grants from the Sigma Xi Scientific Research Foundation, the Agencia Nacional de Promocion Cientifica y Tecnologica (Argentina) a nd the Fundacion Antorchas. The laboratory work was assisted by Jennifer Kelly and Jonathan Auth.

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i Table of Contents List of Tables iii List of Figures iv Abstract vi Chapter One: Introduction 1 Diet as a Key to Understanding Culture 2 The Thesis 3 Chapter Two: Background of the Cuyo 13 Geography of the Cuyo 15 South American Archaeology 18 Research Frame 19 Models for the Adoption of Agriculture 21 Cultural Chronology 23 Paleoindian and Archaic Sites 24 The Holocene Epoch 25 Silencio Arqueolgico 26 Dietary Resources 28 Origins of Domesticated Maize 31 Sample Description 34 Tissue Selection 34 Temporal Distribution 36 Spatial Distribution 37 Sex Distribution 37 Age Distribution 38 Chapter Three: Principles of Stable Isotope Analysis 39 Biological Materials 39 Bones 40 Teeth 42 Skin/Muscle Tissue 44 Hair 46 Carbon Isotope Analysis 46 Nitrogen Isotope Analysis 49 Strontium Isotope Analysis 51 Sources of Error 52

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ii Chapter Four: Stable Isotope Analysis Methods 54 Sample Preparation 55 Bone Apatite 55 Tooth Enamel 57 Bone Collagen 57 Resource Samples 60 Hair 60 Skin and Muscle 65 Instrumentation 67 Chapter Five: Results and Discussion 68 Resource Sample Results 69 Human Hard Tissue Results 72 Bone Collagen 72 Bone Apatite 75 Tooth Enamel 76 Human Soft Tissue Results 79 Hair Samples 80 Skin/Muscle Tissue 85 Spatial Comparison 88 Temporal Comparison 89 Discussion 91 Chapter Six: Conclusion 94 Considerations for Isotopic Studies 95 Scholarly and Educational Significance 96 Applied Perspectives 97 Future Directions 98 References 99 Appendix A 114 Appendix B 116 Appendix C 123 Appendix D 124 Appendix E 125 Appendix F 126

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iii List of Tables Table 1.1 Chronology and site location of human samples 9 Table 2.1 Chronology and associated cultures of San Juan, Argentina 28 Table 2.2 Analyzed samples of floral a nd faunal resources 30 Table 2.3 San Juan samples available for soft tissue analysis 36 Table 2.4 Sex distribution by province 37 Table 3.1 Types of stable isotope analysis for biological samples 43 Table 4.1 Number of hair samples pe r individual 63 Table 4.2 Skin and muscle sample types 66 Table 5.1 Results of analysis of floral a nd faunal resource samples 70 Table 5.2 15N and 13C results from human samples 73 Table 5.3 15N values for hair segments 81 Table 5.4 Students t -test for 15N hair segments 82 Table 5.5. 13C values for hair segments 83 Table 5.6. Students t -test for 13C hair segments 84 Table 5.7 Stable isotope analysis results of skin/muscle tissues 87 Table 5.8 Comparison by provinc e of human bone and tooth samples 89

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iv List of Figures Figure 1.1 Map of South America 4 Figure 1.2 The naturally mummified remains of an infant 8 Figure 2.1 Dry mountain slopes of Argentina 14 Figure 2.2 Mesquite brushlands of the Argentine pampas 14 Figure 2.3 Phytogeography of the Cuyo 16 Figure 2.4 Chinchorro mummy r ecovered from the Atacama Desert of Chile 17 Figure 2.5 Modern-day guanacos, a staple of th e prehistoric Argentine diet 29 Figure 2.6 South American sites with early plant remains 33 Figure 3.1 Three forms of archaeologically preserved bone 41 Figure 3.2 Lateral view of an adult tooth 44 Figure 3.3 Carbon isotope fractionation in te rrestrial plants 50 Figure 4.1 Drilling for tooth enamel sample 58 Figure 4.2 Collagen samples and the chemical solu tions used for processing 59 Figure 4.3 Archaeological samples for stable isotope analysis 61 Figure 4.4 A hair sample representing a twomonth growth period 64 Figure 4.5 Hair samples are placed in tin foil squares 64 Figure 5.1 13C values vs. 15N values for resource samples 71 Figure 5.2 13C values vs. 15N values for bone collagen samples 75 Figure 5.3 13C of bone apatite vs. 15N values of bone collagen samples 76 Figure 5.4 13C values of apatite and tooth enamel samples 78

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v Figure 5.5 13C values for hair vs. skin samples 87 Figure 5.6 15N values for hair vs. skin samples 88 Figure 5.7 13C results for both provinces vary by time period 90

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vi Before the Inca: Prehistoric Dietary Transitions in the Argentine Cuyo Nicole Shelnut ABSTRACT A dietary reconstruction was performe d in order to understand changing prehistoric subsistence patterns in the Centra l Andean geographical area of the Argentine Cuyo that includes the provinces of San Ju an and Mendoza. Archaeologically, the Cuyo is also known as a boundary between Andean agriculturalists and the foragers of Patagonia. One hypothesis being tested is whet her this area was one of the last South American cultural groups to convert to maize cultivation, probably around 2000 BP. The process of stable isotope analysis is used to reconstruct the diet s of individuals, as it reveals the relative proportions of C3 and C4 plants and the cont ribution of aquatic resources to otherwise terrestrial diets, as well as variations in trophic leve l of the foods consumed. In this study the bones, teet h, hair, and flesh from 45 i ndividuals were tested to address specifically total and protein diets, as well as seasonal variation and changes between childhood and adulthood. This process, when used in combination with previous analyses, such as midden or faunal an alysis, allows researchers to evaluate the results of those previous studies, and thus compose a more thorough reconstruction of the lifestyles of a prehistoric culture. Information garnered from this study indi cates that the times of dietary transition were variable, with seasonal patterns becoming more stable over long periods. Furthermore, some members of the study population demonstrate the existence of

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vii nutritional stress indicators, such as dental caries, that can be viewed in relation to the dietary shifts that may have been a cultura l adaptation to the envi ronment of the Cuyo. Overall, this study shows the early adoption of maize agriculture in central western Argentina and recommends future studies that analyze the relationships between agriculture, diet, and nutr ition in the New World.

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1 Chapter One: Introduction This thesis focuses on the interplay betw een human biological systems and their corresponding environments, both natural and cult ural. Stable isotope analysis has been undertaken to examine prehistori c dietary transitions in an ar ea of central west Argentina called the Cuyo. In a broad sense, this thesis examines the ways th at the choices humans make, as members of their dis tinct societies, affect the wa y they live and, in turn, their future descendents. By considering a few remarkable changes in cultural systems as defining moments in the human past, one can begin to explain and possibly predict the wide range of effects that might occur. Transitions such as those from nomadism to sedentism, foraging to farming, and even the relativel y recent occurrences of the Industrial Revolution and globalization can be viewed as a series of even ts that have assisted in shaping the human career. For instance, th e development of transcontinental trade has been associated with a wealth of epidemic s, including the black plague (Magerison and Knsel 2002). Also, the shift from nomadic to sedentary existen ce has been linked to various alterations in artistic expression, craft specializati on, social organization, and the spread of chronic disease (Larsen 2002; Schoeninger and Schurr 1994). Along similar lines, the move from hunter-gat herer to agricultural lifestyles in many populations can be seen as a way in which humans have attemp ted, and in some ways achieved, a control over the natural environments in which they ex ist. However, this transition has not been a one-way process. Along with the adop tion of agriculture, some populations have experienced increases in nutritional pathologies, such as anemia and osteoporosis (Gilbert

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2 and Mielke 1985; Larsen 2002). Pearsall (1994) emphasizes that the introduction of maize agriculture into a society has importa nt biological and cultu ral implications. Diet as a Key to Understanding Culture Although anthropologists have frequen tly used dietary st udies to understand social and economic patterns, researchers ar e now beginning to address the potential impacts upon the human species that result from the transition of hunter-gatherer to agricultural lifestyles. Poor health conditi ons are increasingly being associated with changes in human diet and associated sede ntism (Milton 2002; Sc hoeninger, DeNiro and Tauber 1983). The recently published volume, Human Diet: Its Origin and Evolution begins with similar thoughts: In essence diet is a key to understa nding our past, present, and future. Much of the evolutionary success of our species can be attributed to our ability to procure, process, and cons ume a wide range of foods. However, recent changes in our diet (e.g., increased intake of such things as saturated fat, refined carbohydrates, a nd sodium, and decreased intake of nonnutrient fiber) may lie at the root of many of the health problems swamping our health care syst ems [Ungar and Teaford 2002: 1]. In regard to chronic disease and diet, Ea ton et al. (2002: 12-16 ) outline five ways in which contemporary diets have affected modern human health disadvantageously. These consist of high blood pressure due to ma rked increases in sodium intake; severely reduced consumption of cancer preventing frui ts and vegetables, in exchange for cereal grains, which have demonstrated no evidence of such effects; lack of energy output in exchange for caloric intake; increased dietary fat associated with coronary heart disease; and a relationship between docosahexonic ac id (DHA) deficiencies and reduced brain sizes.

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3 Similarly, the transition to farming ofte n involved an overwhelming dependence on key domesticates such as rice, barley, a nd maize. While these staple crops have allegedly played historic roles in the broad cu ltural systems of societies, reliance on these cultigens can often supersede intake of basi c nutritional resources such as fresh meats, fruits, and vegetables, and result in a diet ary lack of iron and essential amino acids. Maize-based diets are of particul ar interest, as iron, protein content, and niacin absorption are all characteristically low. Lack of these essential nutrients has been linked to increases in anemia and osteoporosis, declin ing oral health, and reduced growth rates throughout various societies (Larsen 1995, 2002; Steele and Bramblett 1988; Ungar and Teaford 2002; White 2000). The Thesis This work provides much needed inform ation that will assist those trying to understand the effects of cultural transitions on human biological systems. It focuses on a range of prehistoric populat ions on the verge of majo r cultural, biological, and subsistence strategy shifts. Dietary reconstruction is used to understand changing subsistence patterns in the Central Andean geographical area of the Argentine Cuyo (Figure 1.1). While the idea of a transition from hunter-gathe rer to agricultural lifeways has been suggested by previous archaeological research, stable isotope analysis provides clearer data on th e area of study. In recent times, researchers interested in studies of prehistoric diets have been limited to mostly indirect methods of investigat ion; for example, the analysis of floral and

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4 Cerro AconcaguaIsthmus of PanamaValdes PenninsulaAmazon BasinA n d e s M o u n t a i n sA t a c a m a D e s e r tMato Grosso PlateauP a m p a sP a t a g o n i a Isla Grande de ChiloeLesser AntillesTierra Del FuegoChile BasinP e r u C h i l e T r e n c hP e r u C h i l e T r e n c hPeru BasinLake TiticacaA m a z o n R .A m a z o n R .A m a z o n R .R i o A r a g u a i aR i o A r a g u a i aR i o J u r u aR i o M a d e i r aR i o M a d e i r aR i o M a d r e d e D i o s Rio NegroR i o O r i n o c oR i o O r i n o c oR i o P a r a g u a yR i o P a r a n aRio ParanaR i o P a r a n aR i o P u r u sR i o S a o F r a n c i s c oR i o T a p a j o sR i o T e l e s P i r e sR i o T o c a n t i n sR i o T o c a n t i n sR i o U r u g u a yR i o X i n g uR i o X i n g uCaribbean SeaGulf of San Jorge Gulf of San Matias Strait of MagellanSOUTH AMERICAAtlantic Ocean Pacific Ocean60W 65W 70W 75W 80W 35W 40W 45W 50W 55W 5N 0 10N 5S 10S 15S 20S 25S 30S 35S 40S 45S 50S 55S 0-150 Ft. 150-300 Ft. 300-600 Ft. 600-1200 Ft. 1200-1800 Ft. 1800-3000 Ft. 3000-4500 Ft. 4500-6000 Ft. 6000-7500 Ft. 7500-9000 Ft. 12000+ Ft. -3000 0 Ft. 500 KM 500 Miles 0 0 Parallel scale at 20S 0E 9000-12000 Ft. Figure 1.1. Map of South America. The geographical region of the Argentine Cuyo is highlighted in red

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5 faunal remains left in middens, cultural iconography, and ceramic use (Keene 1985; Schoeninger, DeNiro and Tauber 1983; Schoe ninger and Schurr 1994). The few methods from which more lucid evidence can be obtai ned include residue or lipid studies of pottery (Charters 1993), coprolite analysis (Reinhard and Bryant 1992), or the very rare incidences in which mummies are recovere d and can be autopsied to discover the contents of their stomachs (Holden and Nez 1993). While many of the indirect methods have proven useful for group r econstructions (e.g., study of Mendoza by Lagiglia 1977), their results ar e in some ways incomplete, as they are only capable of identifying the food sources in a given envi ronment, which individuals may or may not have eaten extensively. Here lies the dis tinction between menu and diet. Among others, Armelagos (1994: 235) clarifies the difference between these two terms; menu refers to the variety of foods available to a population, whereas diet refers to what is actually eaten. The relative importance of these dietary components can be hypothesized by studies of ratios of floral and faunal materi als found in archaeologi cal context, but one can only posit that, in practice, the remains being recovered were actually consumed. In contrast, the diet of in dividuals can be reconstructe d through the use of stable isotope analysis, which reveals the relative proportions of C3 and C4 plants and the contribution of aquatic resources to otherwise terrestrial diet s, as well as variations in trophic level of the foods consumed. During th e last 30 years, stable isotope analyses have enabled researchers to document physical ly the transition from hunter-gatherer to agricultural subsistence patte rns by comparing the proportions of diet represented by resources with differing photosynthetic pr ocesses (Krueger and Sullivan 1984; Schoeninger, DeNiro and Tauber 1983; Schoeninger and Moore 1992; Schwarcz and

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6 Schoeninger 1991; Tykot 2004; van der Merw e and Vogel 1978; Vogel and van der Merwe 1977). This process, when used in combination with other analyses, allows researchers to evaluate the results of previ ous studies, and thus compose a more thorough reconstruction of the lifestyles of a prehistoric culture. In areas such as South America, where a written language did not exist prior to the contact period, stable isotope analysis of human skeletal material becomes an indispensable method of tracking diachronic tr ends in subsistence patterns (Isbell 1997; Pearsall 1992; Schwarcz and Schoeninger 1991) Isotopic analysis can be used to investigate transitions from a subsistence pattern based primarily on hunter-gatherer diets, to intermediary, and eventually maizebased diets. Isotopic analyses of teeth and bone have also been used to understand patterns of food distribution, and compare juvenile and adult paleodiets (e.g., Aufder heide et al. 1994; Cohen 1977; Dupras 2001). One critical limitation of stable isotope anal ysis is that the results of such studies represent ratios of certain food types, rather than actual diet; thus, one might postulate about the relative proportions of maize in an individuals diet as compared to other dietary resources, but they will never be able to give precise lists of what that individual actually ate during their lifetime. Also, researchers are typically confined to the examination of hard tissues, such as bone and teeth, which have far slower rates of decomposition than soft tissue due to the na ture of the materials (Mays 1998). The analysis of the bone samples portrays the averag e diet over the last several years of an individuals life, while that of tooth enamel reflects di et during the age of crown formation. This combination of analyses is useful for contrasting the juvenile and adult diets of individuals, but conclu sions are limited to the representation of average diets

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7 from considerable periods of time (Larsen 1997 ). However, soft tissues have a much more rapid turnover rate. They are particularly advantageous in that the relatively new procedures of isotopic analyses of hair and flesh samples can reveal seasonal, or possibly even monthly dietary variations (OC onnell and Hedges 1999a). The excellent preservation conditions of the materials in this study (Figure 1.2) allow a unique opportunity to study soft tissues, as many are th e result of natural mummification, caused by the arid conditions of the environment in wh ich they were interred at death (White and Schwarcz 1994). This analysis was selected because it had the capability to reflect varying dietary patterns of i ndividuals, and sheds light on prev iously abstract ideas about prehistoric South American lifestyles. Here a dietary reconstruction is used to test explicitly models of seasona l variation, identify any evid ence of ecological stress, and add to scientific knowledge. Further, this work contributes to th e establishment of a dietary baseline in the archaeolo gical record of Argentina. Human samples were provided, along with funding for analysis, by the Museo de Historia Natural, Argentina. The biological samples from 45 individuals range in age from approximately 4100 BP (before present) to 200 BP (Table 1.1). These samples are thought to be representative of the greater human population from multiple prehistoric sites located in the present day provinces of San Juan and Mendoza. These cultural, geographical, and temporal differences allow for the examination of long-term patterns in what is thought to be an area that enc ountered substantial dietary changes around 2000 BP. Gil (2003), among others, has hypothesized that around th is time, many cultures in the surrounding area shifted from hunter-gathe rer to agricultural lifestyles, with an increasing dependence upon maize as the prim ary crop. This thesis evaluates the

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8 hypothesis of dietary change around the time of 2000 BP with questions including: (1) Did a transition from forager to agricultural s ubsistence occur in the Argentine Cuyo? (2) If so, when? (3) Is there a difference be tween the two provinces of San Juan and Mendoza which are thought to have retained separate dietary pract ices in prehistoric times? And (4) what were the effects of this transition if it did indeed take place? As previously mentioned, the samples availa ble for analysis are particularly unique in that many of the individuals were naturally mu mmified. Bone and/or teeth were analyzed for all individuals, while scalp hair, skin or muscle tissue, and/or fingernails were also analyzed for the mummified persons (Appendix A). These materials were processed at Figure 1.2. The naturally mummified remains of an infant, recovered from the province of Mendoza, Argentina

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9 Table 1.1. Chronology and site location of human samples Museum Sample # Province Site Chronology* AF-1083 Mendoza Arbolito 7 100 ENT-2 Mendoza Capiz Alto 400 12 Mendoza Caverna de las Brujas 3850 2038 Mendoza El Desecho AF-2036 Mendoza India embarzada AF-505 Mendoza La Matancilla AF-2000 Mendoza C Negro del Escorial 580 AF-2018 Mendoza Canada Seca 1700-1400 AF-2019 Mendoza Canada Seca 1700-1400 AF-2020 Mendoza Canada Seca 1700-1400 CS-10001 Mendoza Canada Seca AF-508 Mendoza Cerro Mesa AF-510 Mendoza Cerro Mesa 300-200 ENT-3 Mendoza El Chacay AF-673 Mendoza El Manzano AF-13894 Mendoza Gruta del Indio 2300 AF-2021 Mendoza Gruta del Indio 510 AF-828 Mendoza Gruta del Indio 580 AF-830 Mendoza Gruta del Indio 3860 GIRA-27 Mendoza Gruta del Indio GIRA-70 Mendoza Gruta del Indio GIRA-71 Mendoza Gruta del Indio GIRA-831 Mendoza Gruta del Indio JP/J4 Mendoza Jaime Prats 2100-1700 JP-1155 Mendoza Jaime Prats 2100-1700 JP-1352 Mendoza Jaime Prats 2100-1700 AF-8 Mendoza La Olla AF-2072 Mendoza Las Ramadas 970 AF-681 Mendoza Medano Puesto Diaz 2000 AF-500 Mendoza Rincon del Atuel 1760 AF-2025 Mendoza Tierras Blancas 200 AF-2022 Mendoza Ojo de Agua 1200 AF-503 Mendoza RA-1 1760 MGA-1 Mendoza RQ-1 SJ10-ENT1 San Juan Angualasto 600 SJ4-ENT2 San Juan Angualasto 640 SJ5-ENT2 San Juan C Calvario 880 SJ2 San Juan Calingasta 800 SJ6-ENT8 San Juan Gruta 1 MorrillosAnsilta 2000 SJ8-ENT5 San Juan Gruta 1 MorrillosAnsilta 2000 SJ7-ENT2 San Juan Gruta 1 MorrillosAnsilta 4070 SJ1-ENT7 San Juan Gruta Morrillos 7900-4200 SJ9-ENT1 San Juan Hilario 1400-1200 SJ3-ENT3 San Juan Punta del Barro 590 *Given in years BP **Samples submitted for radiocarbon dating, for wh ich the results have not yet been received

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10 the University of South Florida (USF) La boratory for Archaeological Science under the direction of Dr. Robert H. Tykot. Drs. Adolfo Gil and Gustavo Neme, specia lists in the field of South American prehistory, were also consu lted during the various stages of the study. Gil and Neme, archaeologists from the Museo de Historia Natu ral, were present for many of the previous excavations in Mendoza and are involved in ongoing work on the gath ered materials. The following chapters add to scientific know ledge in the fields of archaeology, stable isotope analysis, Argentine prehistory, a nd South American archaeology, while building upon the prior work of Gil, Neme, and their a ssociates. This research can also assist those interested in present-day populations that are struggling w ith nutrition related diseases, such as anemia and osteoporosis. Knowledge of ancient diet patterns may lead to a better understanding of the beginnings of human disease. Stable isotope analysis provides data regarding the transition to agricu lture that can be viewed in relation to diet, nutrition, and disease in both past and present populations. In addition, this research has a public component, in that it provides a greater understandi ng of New World prehistory for both academics and Argentineans. The inte rpretation of the resu lts of this study are incorporated in the exhibits of the Mu seo de Historia Natural, Argentina. The subsequent chapter, Background of the Cuyo begins with a description of the study area and explains the geographical bound aries of the Cuyo. The various cultures acknowledged as previous inha bitants of the environmental zones that make up the region are portrayed. This culture history also di scusses the plant and animal resources that have played important roles in past diet s, and briefly assesses the development of agriculture in the region. Models are confe rred for the origin of domesticated maize,

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11 aspects influencing South American archaeol ogy, and theoretical orientations. Further explanation is given of the rese arch goals with a detailed acc ount of the samples that have been selected for analysis. Next, Principles of Stable Isotope Analysis reviews the fundamentals of stable isotope analysis and its initi al studies. This process is not complete without a clarification of the biological processes a ffecting the development of bones, teeth, hair, muscle and skin tissue. Carbon, nitroge n, and strontium isotope analysis and fractionation are explained. Sources of error that may affect isotope ratios if not taken into account are also considered. This explanatory section enables a greater understanding of the subsequent chapters. The fourth chapter, Stable Isotope Analysis Methods details the various preparation procedures that were used to process the numerous sample materials at the USF Laboratory for Archaeological Science. Laboratory procedures discussed include those that were employed to prepare bone apatite, tooth enamel, bone collagen, scalp hair, skin/muscle, and resource samples. This description also consists of information regarding the instruments of analysis used at the USF St. Petersburg facilities. The Results and Discussion chapter reports the numeri cal figures obtained from stable isotope analysis and the results are examined for accuracy. SPSS 13.0 and Microsoft Office Excel 13.0 were used to pe rform quantitative analysis and create graphical elements. Intra-population relations hips are considered and seasonal changes in diet are examined. Quantitative anal ysis was used to study inter-populations relationships, including tem poral and spatial trends.

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12 Finally, the Conclusion summarizes the work that has been done. Future methodological considerations for isotopic st udies are reported, and a section has been included on the scholarly and educational impor tance this study. Admittedly, this study is but one small segment of ongoing research in the area of interest. The information presented in this thesis is by no means inte nded as an ultimate portrayal of dietary changes and effects in the Cuyo. It is, howev er, hoped that the work that has been done will significantly contribute to understanding of this region and its associated peoples. Chapter 6 specifies the directions of future studies in greater detail.

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13 Chapter Two: Background of the Cuyo The Andes are composed of high peaks, deep basins, low valleys, and rolling grasslands. A diversity of vegetation and climatic zones results, because the Andes are made up of three parallel mountain chains th at were formed by volcanic activity in the Late Cenozoic era (Lamb 2004). These mountain chains are referred to as the Cordillera Principal, Frontal Cordillera, and the Preco rdillera, and are divi ded by tectonic valleys and deep, mountain fed rivers (Compagnucci and Vargas 1998). Interestingly, the seismically active fields of the Andes may have affected both the ways that previous people used their landscapes, and what pres ent day archaeologists know about the sites that remain (Bruhns 1994). In the central Andes, high-altitude lakes, such as Lake Titicaca and Lake Junin, are common. To the east of the mountains, many rivers and valleys flow toward the Amazon, separated by vast grasslands ( punas) and high summits. Moving southward, altitude increases in a variet y of punas and valleys, and then decreases into the coastal deserts of present day Chile along the coast of the Pacific Ocean (Clapperton 1993; Isbell 1997; Bruhns 1994; Willey 1971). The Southern Andes include the majority of present day Chile, southern Bolivia, and western Argentina. The Argentine Ande s are generally dry, and primarily composed of barren slopes, with semi-deciduous scrub forests filling the region (Figure 2.1). Moving south of the La Plata Basin, a terrain of basins and plains, the vegetation changes to pampas of mesquite brush land (Figure 2.2). This area of western Argentina

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14 Figure 2.1. Dry mountain slopes of Argentina Figure 2.2. Mesquite brushlands of the Argentine pampas

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15 encompasses a greater number of mountain streams (Compagnucci and Vargas 1998). As a consequence of the increased access to water, this was the chief area of human occupation throughout prehistoric times, and is often referred to as the southern cone (Bruhns 1994; Pearsall 1992). Geography of the Cuyo The western Andean geographical area of the Argentine Cuyo lines the Central Andes and extends southward beginning with the aforesaid southern cone. The term Cuyo might be based on the indigenous Hu arpe word cuyum, meaning dry or sandy earth (Muoz and Lillo 2001). The area consis ts of three provinces : San Juan to the north; Mendoza to the south; and to a lesser extent, San Luis to the east (Figure 2.3). Although San Luis is politically defined as part of the Cuyo, it is far less dry and mountainous than either the San Juan or Me ndoza provinces, ecological ly dissimilar, and is not thought to have been occupied as thoroughly throughout prehistoric time. The relatively few known sites from San Luis will not be considered in this study. The provinces of San Juan and Mendoza are known for their complex ecosystems, diverse floral and faunal resources, and o ccasional earthquakes. The Cuyo borders the central Andes and sits southeast of the Atacama Desert. It is shadowed by Mount Aconcagua, the second highest mountain peak in the world (6,960 m asl) and experiences somewhat frequent hailstorms. Climate a nd the availability of resources change remarkably over short distances in this region of the world (Gil et al. 2005). The nearby Atacama Desert of Chile serves as an exampl e of one of the most extreme climates of South America, and is the occupation ar ea of the Chinchorro, a well-known population

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16 documented by Uhle (1922). This coasta l group is known for their unique mortuary practices (Figure 2.4), and might have been th e first culture to mummify their infants and children (Arriaza et al. 1998; Aufderheid e 1993; Vreeland 1998). Some (Gil, personal communication, 2004) have even suggested that the Chinchorro in troduced the practice of maize agriculture to the area of the Ar gentine Cuyo; however no substantial studies have been undertaken thus far to address this issue and no archaeo logical remains have been found which would support this argument. The Mendoza-Neuquen region is defined as an environmental and cultural subdivision of the Southern Andes, with an elevation betw een 1200 and 3000 m and extending from 32 to 37 degrees south and 70 to 67 degrees west. This diverse ecological region includes mountai ns, plains, volcanic fields, and deserts. The highlands of the Andes Cordillera are mountainous at 32 degrees south, with eastern piedmonts Figure 2.3. Phytogeography of the Cuyo. Political boundaries of modern day provinces are hi g hli g hted in g reen.

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17 extending toward large plains from 32 to 34 degrees. Melting glaciers and mountain snowfall feed the fluvial systems of the Ro Diamante, Ro Atuel, and Ro Grande throughout the volcanic fields of La Payunia in southern Mendoza at 34 to 37 degrees (Compagnucci and Vargas 1998; Gil et al. 2005 ; Willey 1971). In the past, the area that is now known as southern Mendoza represen ted a unique transition zone between the hunter-gatherers of Patagonia and the semi-sedentary agricultural populations of the Andes. Mendoza is thought to have been one of the last cultural areas to cultivate maize and other prehispanic crops, probably around 2000 BP. Further north, in the province of San Juan, inhabitants were thought to have adopted agriculture somewhat earlier, although still relatively late when compared to the continent of Sout h America as a whole (Gil 2003). The present study employs the method of stable isotope analysis to test those hypotheses; to examine what differences ex isted among contemporary populations of the Cuyo, and to document what dietary transiti ons took place over the broad time span of approximately 6000 BP to 200 BP. It is important to note that sections of the provinces of Mendoza and San Juan are desert environments, with some areas rece iving less than 100 mm of rain annually Figure 2.4. Chinchorro mummy recovered fro m the Atacama Desert of Chile

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18 (Compagnucci and Vargas 1998). This is an ideal environment for natural mummification. Vreeland (1998) defines three types of mummification that occur in the New World: natural mummification caused by environmental factors such as aridity, extreme hot or cold temperatures, and/or lack of air in burial situation; intentional natural mummification, resulting from human exploi tation of said natural processes; and artificial mummification, which is produced exclusively through human manipulation. While the second process, intentional na tural mummification, typifies the cultural practices of residents of the Atacama Desert of Chile, the arid climate of the Argentine Cuyo provides for frequent occurrences of natural mummification regardless of human intervention, thereby preserving soft tissues and hair (Gil et al. 2005). This preservation of soft tissues allows for greater understand ing of dietary complexity in prehistoric systems, including analysis of possible seasonal variation. South American Archaeology Previous archaeological resear ch in South America has a history of strong Marxist orientation (Funari 1997; Politis 1999, 2002). This theoretical perspect ive has resulted in many cultural neo-evolutionary models (Bruhns 1994) that either implic itly or explicitly suggest all cultures have passed through a si milar progression of advancement. South American archaeologists working with prev ious archaeologists records are commonly confronted with neo-evolutionary models a nd many are actively working to develop more flexible and more plausible temporal and chr onological schemes. The research this thesis undertakes is an attempt to develop a regiona lly specific model, while still interpreting

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19 humans as a species that interacts with its environment in a multitude of ways; both influencing and being influenced by its surrounding ecosystems. Research Frame This research fits into a human ecol ogy framework by viewing human subsistence activities as interactions with corresponding ecosystems. Just as environmental variations affect the availability of dietary resources and lead species to adapt to these changes, dietary transitions, such as the adopti on of agriculture in a re gion, can be seen as behavioral adaptations to a surrounding enviro nment. Butzer (1982) argues that human ecology theory applies well to many archaeological studies, as the analysis of human remains and associated artifacts demonstrat e activities that may be perceived as interactions with co rresponding ecosystems. Similarly, stable isotope analysis of hu man remains serves to reflect subsistence activities that may be perceived as reactio ns to, or interactions with, surrounding environments. Stable isotope analysis not only tracks long-term di achronic changes, but the naturally mummified samples that have been analyzed in this study allow for the examination of seasonal dietary variation. The following study documents an intrinsic relationship with the lands where prehistori c inhabitants of the Cuyo subsided, foraged, and later cultivated crops. Gilbert (1985: 340) define s three categories of stre ss that may influence a population or an individual to make change s in their survival techniques: directly environmental, indirectly environmental, a nd psychosocial. While direct environmental changes can include climatic changes, such as those that may have influenced the

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20 inhabitants of the Cuyo in the mid-Holocene (see below), indi rect stresses include disease or nutritional stress also likel y products of environmental ch ange. The third category of stress, psychosocial, includes pressure from ot her members of an indi viduals culture. Here, it is important to recognize the c oncept of human agency in a decision making process (Dobres and Robb 2000). Again, one does not want to fall prey to the sort of determinism that plagued stage theorist s such as Morgan; in reality, the possibility exists that no action at all may be taken in re sponse to any of the stress sources indicated above. Conversely, adaptations can be made either consciously or subconsciously. Certainly, adaptations to stress can be biological, but they can also be behavioral. The concept of human agency is epitomized in the Cuyo. Not only might some cultures have chosen to take on maize agri culture (e.g., the inhabitants of San Juan), others (e.g., the inhabitants of Mendoza) may have remained foragers well up until the arrival of Spanish conquistadors. Additionally, not all intentional adaptive de cisions necessarily re sult in biological or cultural success (Gilbert 1985) Intentional alterations in diet may be unintentionally maladaptive. Many anthropologists (Larse n 1995, 2002; Steckel et al. 2002b) have pointed to detrimental changes in health that correspond to th e adoption of maize agriculture, such as anemia and osteoporosis. Moreover, some (e.g., Steckel et al. 2002a) note agricultural societies were the first to fa ll victim to the health epidemics brought on by the arrival of Spanish conquistadors. This suggests that the success of the European invasion might have been due to weakened immune systems of natives brought on by inadequate diets, thereby making indigenous cultures such as the Inca easy prey for European disease vectors. Furthermor e, some members of the study population

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21 examined in this thesis demonstrate the existe nce of nutritional stre ss indicators, such as dental caries, that can be view ed in relation to the dietary shifts that may have been a cultural adaptation to the envi ronment of the Cuyo. For thes e reasons, it is important to question not only when dietary transitions occu rred, but also what the impetus might have been for these changes, and how they may have affected the popul ations that undertook them. Models for the Adoption of Agriculture Multiple competing models accounting for the adoption of agriculture have been proposed (see Steckel et al. 2002a for a thorough review). Many such models interpret the trans ition to food production in va rious regions of the world and can be grouped under the broad heading of evolutionary ecology. One such perspective (Price and Gebauer 1995) argues that populations made very conscious decisions to adopt agriculture as a sort of reserve method in otherwise satisfactory ecological systems. Thus they adopted primary food production because they could afford the risk. Assuming this gamble was successful, increased crop quantities supported larger population sizes, th ereby allowing for the development of increasingly comple x political systems such as the preHispanic Inca Empire. Another interpretive stan ce incorporates the view that humans were forced to begin practicing agriculture, because of envir onmental scarcity or nutritional deficiencies (Gilbert 1985). On a regiona l scale, climate changes during the Holocene epoch are thought to have lessened the availability of protein resources in the Cuyo when large

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22 game became extinct as grazing lands diminish ed. This may be one reason some cultures eventually decided to adopt ma ize agriculture, while others chose to hunt smaller animals and continue to gather wild pl ants. It has also been sugge sted (Cohen 1977; Steckel et al. 2002a) that population increases in the Neolithic took place prior to the adoption of agriculture, thereby providing a type of social stress, in th e form of population pressure that was combated with the behavioral adap tation of landscape modi fication in the form of agricultural systems. Thus, food produc tion might be viewed as an attempt to compensate for some sort of perturbation in a cultures nutritional system. Keene (1985) points to dietary shifts as an indicator of nutritional st ress, noting that many huntergatherer groups were completely capable of sustaining and even exceeding their energy needs in appropriate environments. Keene vi ews the transition to agriculture as a risk that would only be undertaken in otherwise unbalanced or insufficient ecosystems, that were, for some reason, suddenly incapable of supporting human populations. Price and Gebauer (1995) further illu minate these alternating viewpoints by grouping the potential motivations for adopt ion of agriculture into exogenous and endogenous factors. Endogenous factors include social change and allude to a conscious alternation of subsistence st rategies. Exogenous influences include climate changes and population pressure, thereby viewing the a doption of food production in response to environmental perturbations. They, too, sugge st that agriculture was first adopted in areas of abundant resources; however, they reason that significant population is a condition for, rather than a cause of, food production. Their global analysis indicates no presence of remarkable population growth im mediately prior to a dietary transition.

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23 Some of the cultural changes that are t hought to have taken place in the provinces of San Juan and Mendoza are examined below. A causal relationship between environmental/social stressors and cultural changes is not assumed; however, it is important to acknowledge these transitions so that any existing correlations may be taken into account and examined in further detail. Cultural Chronology In general terms, little is known about Sout h American prehistory when compared to the rest of the New World, or most certainly to the Old World. In part, this may be due to the lack of an indigenous writing system. Native Americans of the Andes did not have writing systems as are known today, and th eir knotted string records, called quipus may never be fully understood (Isbell 1997; Urton 2003). Quipus are thought to have been a Huari invention that served as mnemonic devices for the Incas class of scribes, called the quipucamayoc (Bruhns 1994). While archaeol ogists (Quilter a nd Urton 2002) now recognize the numbering system that these kno tted strings might represent, the meaning of any individual quipu is not known (Urton 2003: 161-164), and their translators are now extinct without having passed down their specialized knowledge, due in part to the invasion of Spanish conquistadors. Furthermore, the research that has been undertaken in South America has predominantly focused on information from Peru, or Inca populations (Pearsall 1994; Politis 1999, 2001). To this effect, relatively little is known about Argentina. While it might be helpful to examine changes in the Cuyo compared to broad continental movements, few horizon lines cultural chr onologies defined by distinct artifacts

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24 indicative of the rapid spread of a culture over a wide period of time exist for South America in general. Experts commonly define the four central Andean horizons as Inca, Tiahuanaco-Huari, Cuzco, and Chavin (B ruhns 1994; Vreeland 1998). While this information is spatially consistent with the area of the Cuyo, the full prehistory of Argentina has not been considered in detail nor is the temporal span particularly relevant. There exists a predominance of information regarding Inca sites, with few previous reports centering on the time period being consideration in this study. For this reason, continued research in Argentina is especially important and should include, but not be limited to, the scope of this thesis. Paleoindian and Archaic sites It is now recognized that areas of South America, such as the pre-Clovis site Monte Verde in present-day Chile, were occu pied at least 12,500 y ears ago, with later sites occurring much more frequently (Dill ehay 1999). Thus far, little is known about South American inhabitants east of the Andes mountain range. However, finds throughout northeast Argentina, Paraguay, and Uruguay suggest the region was occupied throughout the early Holocene (Gil et al. 2005 ; Bruhns 1994). The information that does exist includes the analysis and interpretation of Archaic sites including Ayampitn and Inithuasi, with the Ayampitn tool industry reco gnized as a distinct style of early points and flakes. Named after the site of Ayampit n, an open air camp in Cordoba (to the north of the Cuyo), this grouping includes distinct ive willow leaf points, flakes, and grinding tools. While tools of this type have been recovered from a number of sites, the most

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25 well-known assemblage is from Intihuasi Cave (Gonzlez 1959), because the clearly stratified deposits make re lative dating possible. Another early tool industry is the Ampajango, first id entified in the northwest Argentine province of Catamarca (Bruhns 1994). The Ampajango tool industry is primarily defined by a group of percussi on flakes; however, disagreement exists regarding the temporal scheme of these tools. While some have argued for an earlier date because tools of this industry are made w ith minimal modification and lack projectile points, Bruhns (1994) states th at this general lack of sophi stication is due to the poor quality of rock available in th e area, and not some sort of in competence by tool makers of the early Holocene. The Holocene Epoch The progression from the Late Pleistocen e to the early Holocene is known for worldwide perturbations in climatic and e nvironmental conditions. These global changes might have placed environmental stress on human populations, and thereby contributed to dietary transitions. While climates vari ed widely, many areas experienced increased temperatures resulting in a diminution of gr asses, decreased grazing lands (Sandweiss et al. 1999; Zrate 2002), and the eventual exti nction of some large mammals, notably the ground sloth (Long et al. 1998). These climat e changes were particularly pronounced throughout the Andes, resulting in a number of environmental changes that probably affected the early inhabitants of Argentina. Higher snowfall in the upper Andes led to neoglacial advances, resulting in decreased wa ter flow in the major rivers of the Cuyo that are ordinarily fed by melting snow and ice. Meanwhile, fewer summer rains at the

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26 piedmont level probably led to a concentra tion of human populations along large fluvial valleys. Further, these changes directly aff ected the availability of floral and faunal resources throughout the Cuyo. Intensity a nd location of human occupations changed, with lowland sites most likely abandoned. Gil et al. (2005) hypothesize that these environmentally stressful c onditions brought on by decreasi ng water resources in the already dry environment of the Cuyo, led to human responses of a movement upland, where the increased snowfall provided for comp aratively greater water availability. Silencio Arqueolgico The aforementioned occupational hiatus of lowland Argentina is suggested by a lack of sites from this period, and corres ponding low densities of materials (Gil et al. 2005). Alternate explanations such as disturbance by m odern populations and site formation processes, have been examined w ith negative results (G ambier 2000). It is believed that as environmental conditions became increasingly dry from the Late Glacial to the mid-Holocene, many South America populations declined in number and/or adapted culturally. The period of 9000-4500 BP is sometimes referred to as silencio arqueolgico, or archaeological silence, in the most affected areas (Gil et al. 2005; Nuez et al. 2001). This occ upational hiatus likely took pl ace in the most affected environments of southern Mendoza. Essentia lly, environmental perturbations influenced human populations to abandon previous occupati on sites and relocate during this period. Alternatively, Lagiglia (2001, 2002) has propos ed models of continuous occupation in southern Mendoza throughout this time.

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27 Gil et al. (2005) tested this hypothesi s by analyzing 97 radiocarbon dates from the time interval of 14,000 to 200 years BP. Af ter examining these carbon-14 dates they found that the archaeological record does not suppor t Lagilias (2001, 2002) earlier assumption of a continuous occupation in sout hern Mendoza. Rather they agree with a significant hiatus during the period of 7000-6000 BP. Further, they cite the aforementioned geomorphological processes of the Holocene as the impetus for this cultural trend. While less is known about the early cultu ral chronology of San Juan (Table 2.1) than Mendoza, the archaeological record is currently being examined, and this thesis will contributes to the construction of a more thorough cultural chronology based, in part, on cultural transitions such as dietary shifts. Moving toward the late Holocene, rain fall increased again and major fluvial systems were once again hydrat ed. Humans began to move back into the Cuyo, while simultaneously modifying their subsistence stra tegies to adapt to the new environments they were living in. It is emphasized that th e extent of the silencio arqueolgico varies with the scale of spatial analysis. Again, the most climatically affected areas of San Juan and Mendoza are thought to have been abandone d for the longest amount of time, with less fragile environments being reoccupied earlier. The oldest recorded cultigens in Mendoza occur in a funerary cont ext as early as 2200 years B.P., at sites such as Gruta del Indio. Subsequent indicati ons of agriculture from various archaeological contexts do not come until 1,000 or more years later. This information has been used to argue that maize may have had a special cultural mean ing before becoming a dietary staple (Gil 2003). Disparities in the adoption of maize agriculture also occurred in different

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28 environmental regions, such as the Argentine highlands and areas located to the direct east of the mountains. This thesis tests th ese hypotheses, questioni ng when, if at all, maize was adopted and in what areas. While Mendoza and San Juan are sometimes considered separately with San Juan thought to have converted to agriculture earlier than Mendoza this th esis tests this hypothesis with qua ntitative data obtained from stable isotope analysis. Dietary Resources Although the Cuyo is a dry, desert-like e nvironment, it is also known for its diversity of floral and faunal resources (G il et al. 2005). Certainly, the abundance of available dietary resources was utilized by previous inhabitants of the Cuyo, and may have led to the sustained trad ition of hunter-gatherer subsisten ce patterns in the province of Mendoza. Dietary resources that are consid ered to have been particularly important prior to, and in addition to, maize include gua naco (Figure 2.5), rhea, squash, and various fruits. Table 2.1. Chronology and associated cultures of San Juan, Argentina Chronology (BP) Cultural period Cultural group 8500-8200 Early hunter-gatherer La Fortune Industry 7900-4200 Late hunter-gatherer Los Morrillos 3800-1950 Early farming Ansilta 1950-1400 Early agropastoralists P unta del Barro cultural phase 1400-1200 Early agropastoralists Calingasta 1200-900 Middle agropastoralists La Aguada influence 750-460 Late agropastoralists Angualasto/late Calingasta 460-420 Inca Inca, local group with Inca influence 420-388 Local indigenous Huarpes/capayanes and yacampis

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29 While this thesis primarily focuses on the analysis of human samples, it is important to acknowledge the available resour ces humans may have also been eating. A growing number of South Amer ican archaeology projects are incorporating botanical recovery methods (e.g. Barberena 2002; Hastorf 1999; Hastorf and Johannessen 1994; Pearsall 1992) into their research and this has resulted in an exponential increase in knowledge of past dietary options (Bruhns 1994) In the Cuyo, a registry of carbon and nitrogen stable isotope values for alleged di etary staples is being built. Table 2.2 lists floral and faunal species that were analyzed to this effect. The analyses of resource samples will define dietary values in an attempt to rule out the possibility of any abnormal carbon or nitrogen results that ma y affect human values irregularly. South American archaeologists are also looking towards advanced laboratory methods to document the actual inception of maize agriculture. The crop that may have had the greatest effect in Argentine prehistory is maize ( Zea mays ). As discussed in the previous chapter, the adoption of food production has been associated with a multitude of Figure 2.5. Modern-day guanacos, a staple of the prehistoric Argentine diet

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30 cultural, political, and biologi cal changes (Gilbert 1985; Steckel et al. 2002). Therefore, this thesis focuses predominantly on the history of this particular crop in the study area. Prior to the use of stable isotope analysis, archaeologist s struggled to evaluate the transition to maize agriculture throughout Sout h America; without any type of chemical analysis, researchers often had to settle for th e few artistic depictions of maize that exist on limited numbers of ceramics and sculptur es (Bruhns 1994; Pearsall 1994). This proved problematic for a number of reasons including that few cultures produced such stylistic renderings, and thos e that did (e.g., the Inca) were relatively late. Another indirect method of documentation that was us ed in the central and southern Andes was the mere presence (or absence) of manos and me tates or other grinding materials. Bruhns (1994) has argued that this method of interp retation is highly inaccurate, as many South American people either did not grind corn, or did not use manos and metates to do so. Table 2.2. Analyzed samples of flor al and faunal resources Latin Name Common Name Cassia arnottiana Cassia Chaetophractus villosus Armadillo Chenopodium sp. Chenopodium Cholephaga melanoptera Andean goose Cucurbita maxima Winter squash Geoffroea decorticans Chanal Lagenaria sp. Gourd Lagidium viscacia Chinchilla Lama guanicoe Guanaco Phaseolus vulgaris Common bean Prosopis sp. Prosopis tree Pterocnemia pennata Lesser rhea Rhea americana American rhea Schinus polygamus Pepper tree Zea mays Maize

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31 Beyond this, the subject of how and when maize species reached South America has previously served as a subject of much disagreement. Origins of Domesticated Maize In general, there exist three competi ng schemes regarding th e origins of maize domestication. Beadle (1980) and Galinat (1983) have argued that a wild grass, called teosinte, is the ancestor of a ll maize variations. This sugge sts a single origin of maize, that must have traveled to spread across a ll of the Americas. Alternatively, Mangelsdorf (1947) presented a model that contends for multiple and separate origins of maize, including teosinte, from wild tripsicum gra ss in multiple environments. This position also allowed for an independent center of or igin in the central A ndes. Finally, in 1983, Iltis presented a radical theory that asserts an almost inst antaneous evolution of maize with the sexual reversal of the male tassle of teosinte grass. This alteration could occur in a number of manners, including random mutati on, environmental disturbance, or human domestication. The final two models are made more plausible by the observation that South American corns possess rather dissimilar ch aracteristics from Mexican and Central American corns. Bruhns (1994) draws atten tion to this divergence in shape and size, arguing that all flour corns orig inated in South America. However, Pearsall (1994), an expert in the topic of maize domestication, favors a single origin. A common explanatory theme in the multiple origin models is the spread of maize seeds by migratory animals such as birds. Understandabl y, Pearsall sees these as arch aeologically unve rifiable and notes it is plausible that several varieties of maize may have origin ated and subsequently

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32 vanished in the past. Because th e temporal scheme of this thesis is significantly later than the time at which this would be of major concern, the more common interpretation of the origin of maize will be assumed. In a ddition, one could refer to Pearsall (1994) for further exploration of this topic. It is now generally agreed that maiz e was domesticated from teosinte in Mesoamerica between 10,000 and 5,000 B.C. Long and colleagues (1989) used accelerator mass spectrometry (AMS) to show th at the earliest maize remains are at least 4700 years old. Benz and Long (2000) later used morphological an alysis and genetic tests on Mesoamerican maize to argue that the domestication of maize might have occurred even sooner, perhaps earlier th an 5400 BP (Benz 2001). The species then moved through Central America into South Amer ica, eventually adapting to elevations of 3,000 m in the Andes (Bray 2000; Pearsall 1994). The first scientific discoveries of preceramic maize in the Andean region was made by Willey and Corbett in 1941 and 1942 (Willey 1953), however, it took researchers many years to grasp the importance of these discoveries. It was not until the 1950s when Lanning broke new ground with his research in the central and north-central coastal areas, that knowledge of pre-ceramic maize in the Central Andes truly became a focus of Ne w World archaeology (L anning and Patterson 1967). Today, there is a mass of information pertaining to maize from prehistoric Peru, identified from the southern Andes, in cluding information regarding Argentinean agriculture (Bonavia and Grobman 1989; Isbe ll 1997; Keene 1985). Furthermore, maize may have been utilized in diffe rent ways throughout the Andes, and not every culture that

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33 as well as other South American culture areas (Figure 2.6), but sti ll little has been Figure 2.6. South American sites with early plant remains. From Pearsall (1992:176)

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34 knew of maize necessarily adopted it (Pearsall 19 94). For these reasons, it is important to look at the adoption of maize agriculture on a regional scale, such as the study area of the Argentine Cuyo. Sample Description Adolfo Gil, Gustavo Neme, and Humberto Lagiglia, of the Museo de Historia Natural, San Rafael were primarily respons ible for the collecti on of archaeological materials throughout Mendoza. Samples were se lected in an attempt to define site locations and to construct a broad temporal and spatial inventory of the Argentine Cuyo, with an emphasis placed on sites with hi gh levels of preser vation and little postdepositional disturbance. This thesis continues recent exploratory work from the province of San Juan, with many samples being selected from current museum collections. The excellent pres ervation of the museum sample s aids in the experimental analysis of soft tissues materials. The majo rity of human samples that have already been processed are now in the Museo de Hist oria curation, with a minimal amount of processed material remaining at the Univ ersity of South Florida Laboratory for Archaeological Science. Tissue selection Bones and teeth are the most frequently pr eserved biological materials, other than shells and charcoal, in any given archaeological context, as they have much slower rates of decomposition than soft tissue. Bone is the most common of all tissues, and is composed of an organic matrix formed by co llagen, which is sometimes referred to as

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35 gelatin; and an inorganic portion termed hydroxyapatite which is commonly called apatite (Armelagos 1994; Boutton 1991; Sc hwarcz and Schoeninger 1991). Collagen represents a large fraction of bone (approximately 20 to 25 percent in fresh bone), is relatively insoluble, and contains both nitr ogen and carbon. Apatite represents 75 to 80 percent of fresh bone, but contains no n itrogen (DeNiro and Schoeninger 1983). The analyses of the bone matrices, collage n and apatite, portray the average diet over the last several years of an individuals li fe, while those of toot h enamel reflect diet during the age of crown formation. This combin ation of analyses is useful for contrasting the juvenile and adult diets of indivi duals, but conclusions are limited to the representation of averag e diets. The materials in this study provide a unique opportunity to study soft tissues, as the arid conditions of the environment where they were interred often result in natural mummification. Soft ti ssues have a particular advantage, in that the relatively new procedures of isotopic anal ysis of hair and flesh samples may reveal seasonal, or even monthly, dietary va riations (OConnell and Hedges 1999a, 1999b). While the arid conditions of the Cuyo do provide for some natural occurrences of mummification, there was unsurprisingly a pred ominance of hard tissue available for this study. Whenever possible, all available materials were analyzed for each individual, including both soft and hard tissues. Soft ti ssues that existed for analysis included scalp hair, fingernails, skin and muscle, all of which originat ed from individuals of the San Juan province (Table 2.3).

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36 Temporal distribution Radiocarbon dates have been obtained for sites throughout bo th provinces of study whenever possible (Gil et al. 2005). This method of dating helps establish chronology that is otherwise determined thr ough indirect analysis of archaeological materials such as pottery and tool typologies Human remains that have been analyzed range in antiquity from approximately 6000 to 200 BP. This wide temporal span encompasses a number of cultural transitions th at may also represent dietary shifts (see Table 2.1). This thesis seeks to evaluate the changes from h unter-gatherer to agricultural and pastoral societies that have been assumed for the province of San Juan, and to determine if these transitions are reflected in the dietary record of the Mendoza province. The stable isotope analyses reported here co mpare variation for these periods from both provinces. Table 2.3. San Juan samples available for so ft tissue analysis (Gil, pers. comm.) Sample Site Sex Age Chronology (years BP) Hair Skin/ MuscleNail SJ1-ENT7 Gruta Morrillos F 7900-4200 X SJ2 Calingasta 800 X X SJ3-ENT3 Punta Del Barro 590 X X X SJ4-ENT2 Angualasto 640 X SJ5-ENT2 Calvario 880 X X SJ6-ENT8 Gruta MorrillosAnsilta F Adult 2000 X X X SJ7-ENT2 Gruta MorrillosMorrillosM Adult 4070 X SJ8-ENT5 Gruta MorrillosAnsilta M 2000 X X X SJ9-ENT1 Hilario F Adult 1400-1200 X X X SJ10-ENT1 Angualasto F 600 X X X

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37 Spatial distribution Human and resource samples were sele cted from sites throughout both provinces, with the majority of samples originating fr om the Mendoza province. A total of 27 sites were sampled, 21 of which were located in Mendoza and six from San Juan. Mountainous, piedmont, and lowland sites were sampled to gather information from a wide variety of environments (Gil et al 2005; Neme 2002). Many of the sites are rockshelters or situated al ong banks of the major rivers that run throughout the dry environment of the Cuyo. It is emphasized that work has only just begun in San Juan, and the analysis that is presented here just be gins the effort that will be undertaken to document the prehistory of the San Juan province. Sex distribution Sex determinations were made on the ba sis of morphometric analysis performed by Barrientos, Perez, and Novellino (unpublished data). The selected samples appear in Table 2.4. Whenever possible, human sample s were assigned to male or female categories; unfortunately, a large number of skeletons were incomplete, juvenile or Table 2.4. Sex distribution by province Province San Juan Mendoza Female 47 Male 27 Unknown 421 Total 1035 Sex Ratio 2:1:21:1:3

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38 otherwise found indeterminable, and were si mply labeled as unknown. Whereas only individuals of known sex were used to test sex differen ces, individuals of unknown sex were also analyzed to evaluate whether thos e individuals had simila r dietary patterns to those of known sex. Age distribution Barrientos, Perez, and Novellino (unpublis hed data) were also responsible for determining the ages of individuals base d on morphometric and dental analyses. However, many individuals coul d not be assigned ages, and th ose that were assigned age groups presented a discontinuous sequence. Fu rthermore, while some individuals from Mendoza were determined to be children, very few juveniles were available from either province. Therefore, dietary differences re garding age distribution were analyzed only on the basis of individuals apatite to enam el comparison, which yields information on adult and juvenile diets, resp ectively. In order to furthe r understand these processes, the following chapter discusses the principles of stable isotope analys is, including isotope fractionation, fractionation between trophic leve ls, and sources of e rror that may affect isotope ratios.

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39 Chapter Three: Principles of Stable Isotope Analysis The science of isotopic analysis repres ents a fusion of two diverse fields, biomedical research and archaeology (Sullivan and Krueger 1984). Procedures originally developed for radiocarbon dating have give n researchers the ability to examine the chemical makeup of biological material a nd examine the human past in a direct, quantitative manner. This practice serves as an excellent example of the holism of the field of anthropology and is founded on the pr inciple that nearly all biochemically significant elements exist as a mixture of two or more isotopes with differing numbers of neutrons, and identical numbers of electr ons and protons (Schwarcz and Schoeninger 1991). Methods of stable isotope analysis have been defined fo r elements including carbon (C), nitrogen (N), and strontium (Sr). Researchers use mass spectrometers to measure the isotopic ratios of a sample. These results are then compared with the isotopic ratios of a standard. The results yi elded from this type of analysis may speak volumes about diet, nutrition, and migration pa tterns of otherwise prehistoric cultures (Eaton et al. 1988; Ericson 1985; Fogel a nd Tuross 2003; Larsen 2002). Carbon and nitrogen isotope analyses are often used to reconstruct past foodways and the information garnered can then be extrap olated to supplement dietary studies, whereas strontium analysis is often used to examine the hom e range of an individual and extract data regarding relocation or migration affairs. Biological Materials The adult human skeleton is composed of approximately 206 bones and accounts for approximately 14 percent of a living pe rsons body weight (Steele and Bramblett

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40 1988). As briefly discussed in the introductory chapter, the i nherent properties of hard tissues (bones and teeth) allow for greater pr eservation in the archaeological record than their counterparts, the soft tissues. As a re sult, the majority of stable isotope studies performed thus far have used bone and/ or tooth materials for analysis. The exception to this generalization is an extreme environment. Climates with anaerobic conditions, hyper-aridity, or freezing temperatures often allow for higher levels of preservation (OConnell and Hedges 1999a). The present study is un ique in that many of the human samples originated from a dese rt environment that runs parallel to the Argentine Andes. Thus, soft tissue materi als (e.g., hair, skin, muscle tissue, and fingernails) were available for a proporti on of the study population. This range of materials allows for greater understanding of prehistoric diet in the Argentine Cuyo, as well as advances in isotopic studies as a whole. Prior to e xplaining the chemical processes involved in such analysis, a brief discussion of the composition of human biological materials aids comprehe nsion of the study as a whole. Bones The two complexes of bone, collagen a nd apatite, compromise for bone stress from different activities, su ch as tension, compression, and bending. Collagen, the more flexible portion formed primarily from pr otein, allows bone the ability to bend and compress; a collagen deficiency, such as th at caused by nutritional maladies, may cause brittleness in bones that make them more ap t to fracturing. Alternatively, apatite is composed of all dietary components and prot ects bone from compre ssion. Dietary or

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41 nutritional deficiencies, partic ularly a lack of calcium, ma y cause a demineralization of bone, decreasing the ability to compensate fo r body weight. Bones may bend and flex in excessive ways without the support of appropr iate amounts of apatite. An example of this condition is rickets, a condition commonly experi enced by children who lack sufficient amounts of vitamin D (Larsen 1997; Steele and Bramblett 1988). Early stable isotope studies, such as those by van der Merwe and Vogel in 1977 and 1978, focused on collagen, the organic portion of bone. However the inorganic portion, apatite, is less suscepti ble to deterioration than co llagen. Methods were soon developed to extract apatite for analysis (Sul livan and Krueger 1981). Apatite analysis is Figure 3.1. Three forms of archaeologically preserved bone. From left to right: complete bone, bone collagen, bone apatite.

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42 somewhat disadvantageous in that it does not allow for nitrogen studies (Table 3.1), but it may be resorted to when preservation is less than ideal, and is certain ly useful for a more reliable depiction of the preh istoric diet as a whole (Schoe ninger et al. 1983). Both complexes are susceptible to the constant decomposition and regeneration of bone that is the work of osteoblast, osteocyte, and oste oclast cells (Ortner a nd Putschar 1981); this results in an artifici al homogenization of stable isot ope values throughout the period it takes for each complex to recycle itself. Thus stable isotope values obtained from bone samples represent the average di et of the last few years, approximately seven to 10, of an individuals life (White 1993). Teeth Samples available for this study included teet h from individuals of varying ages and it is important to note the differences between juvenile and adult den tition. The juvenile dental set is comprised of 20 deciduous teeth. These milk teeth are subsequently replaced by the permanent teet h throughout childhood (White 2000). The adult dental set is typically compri sed of 32 teeth, subse quent to variation based on factors such as numb er of third molars. Each tooth is bound to the jaw via a periodontal ligament surrounded by cementum (F igure 3.2). Cementum is a somewhat softer tissue than the other materials co mposing teeth and is often lost in the archaeological record. Extending from the root s of the teeth upwards thru the bulk of the crown is a hard tissue called dentin that is a pproximately 30 percent organic. A layer of

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43 enamel then superimposes the entire structur e (Steele and Bramblett 1988). It is this portion of the tooth that is most co mmonly sampled for isotopic analysis. The composition of tooth enamel is similar to that of bone apatite, but teeth do not recycle themselves once they are formed. Thus, stable isotope values obtained from tooth samples represent diet at the time of formation. Te eth form at different times throughout an individuals life, with third molars typically developing sometime between 10 and 12 years of age, and other tooth type s forming between zero and two years (Mays 1985; White 2000). Juvenile values might even reflect the diet of their mother, from either the period in the womb or from th e nursing period (Dupras 2001). Alternatively, adult dentition reflects the values of corres ponding juvenile diets th at will vary in age depending on the tooth type. Table 3.1. Types of stable isotope an alysis for biological samples Element Form of results Material Complex Turnover rate Carbon 13C Bone Collagen 7-10 years Apatite 7-10 years Tooth Enamel Variable depending on time of formation Skin/muscle Collagen Variable depending on depth Hair Collagen > 12 days Nitrogen 15N Bone Collagen 7-10 years Skin/muscle Collagen Variable depending on depth Hair Collagen > 12 days Strontium* Sr87/Sr86 Tooth Enamel Variable depending on depth *Not analyzed in this study

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44 Skin/Muscle tissue White and Schwarcz (1994) demonstrated the utility of soft tissues to document shifting seasonal dietary patterns in Nubian mummies. Subs equent studies, such as those by OConnell and Hedges (1999a, 1999b) and by Fenner (2002), have built upon their work and supported the value of such studies but soft tissue analysis is still fairly uncommon. Whereas hard tissue studies can be useful for determining average diets over the last few years of an individuals life, so ft tissues experience continuous regeneration (Uzuka and Sakamoto 1967). This is partic ularly advantageous for the study of shortterm, or even seasonal, diet. Variation in growth is linked to nutritional stress and physiological dynamics, but is not easily infl uenced by aspects such as environmental Figure 3.2. Lateral view of an adult tooth. From Steele and Bramblett (1988:72).

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45 season or equatorial zone (White 1993). So ft tissues include ha ir, skin and muscle tissues, and finger and toe nails. Skin is composed primarily of collage n, but also contains lipids and the protein keratin. Factors including irre gular, and therefore unpredicta ble, depletion of lipids and amino acids may bias isotopic values when soft tissue begins to decay (OConnell and Hedges 1996; White and Schwarcz 1994). Thus, a lipid extraction was performed on soft tissue samples prior to mass spectrometric an alysis in an effort to ensure greater reliability of results. Alternatively, muscle is composed almost completely of protein and is often left unprocessed prior to analysis, if any analysis does indeed take place. Not only were muscle samples analyzed in this st udy, they were also processed to remove any possible contaminants prior to analysis (Wh ite and Schwarcz 1994). This lipid extraction will be discussed in greater detail in the subsequent chapter. The depth of skin is sometimes difficult to determine in mummified individuals and it is quite possible to conf use desiccated muscle and skin when obtaining samples. It should be noted that while there are major di fferences between the composition of muscle and skin tissue, the researchers who took th e samples from the Argentine population are not entirely confident in their differentiati on between the two types. While this is unfortunate, it would be very precarious to forge ahead assuming all samples were properly assigned to their categories when re searchers have expressed some doubt. Thus, skin and muscle samples are discussed togeth er throughout the remainde r of this thesis. While results of analyses are discussed, the main benefit of this particular portion of the study is intended to be the contribution of additional knowledge of laboratory methods for skin and muscle processing where the scientific record is lacking.

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46 Hair Hair, much like muscle, is composed of almost pure keratin protein. It is exceptional for tracking short-term dietary change s due to its constant generative state. Samples taken along the shaft of the hairs length correspond to diet prior to an individuals mortem, with the area nearest to the scalp yielding stable isotope values representative of the pe riod closest to death. Scalp hair typically grows from .8 to 1.4 cm per month, with an accepted rate of 1.1 cm per month, or .35 mm per day, consid ered standard. Lacking a naturally monotonous diet, the isotopic values of hair samples reflect variation within 12 days, with isotopic equilibrium attained in an extr a two to five months. Jones et al. (1981) caution researchers that the use of whole strands for analysis can confuse results, as older values will mingle with newer ones. For thes e reasons, a 20 mm section is used regularly to analyze carbon and isotope values for two-month periods (Fenner 2002; White 1993). Carbon Isotope Analysis Little is known about the initial acceptance of maize into the human diet although it has long been recognized as a staple crop in South America. The importance of maize in relevance to other foods can be measured by analyzing a ratio of the content of two stable isotopes of carbon, 12C and 13C, in human bones (Fogel and Tuross 2003; Pearsall 1994; Schoeninger and DeNiro 1982; Schoe ninger and Schurr 1994; Smith 1995; Sullivan and Krueger 1981); this ratio reflects the isotopic ratios of the foods eaten by an animal, and statements can be made concer ning the implementation of increasing levels of maize into prehistoric diets, and subs equently, the history of agriculture.

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47 Carbon occurs in th ree isotopic forms: 14C, 13C and 12C. Carbon 14 is the carbon isotope used for radiocarbon dating; it decay s (at a known rate) and is therefore known as radioactive. 13C to 12C are carbons stable isotopes (van der Merwe 1982). The ratios of 12C to 13C vary among plant communities, atmo spheres, and trophic levels. Mass spectrometric techniques can measure these ra tios of carbon isotopes in a given plant, e.g., maize (Boutton 1991; Schwarcz and Schoe ninger, 1991). It is this variation among carbon isotope contents in plants that allows researchers to asse ss the importance of specific food products in an individuals diet. Although there are three photos ynthetic types, most plants consumed by humans can be grouped into one of two terrestrial types: C3 plants use the process known as Calvin-Benson, and convert carbon dioxide into a phosphoglycerate with three atoms during photosynthesis; and C4 plants enable a photosynt hetic pathway called HatchSlack, and convert carbon dioxide into di carboxylic acid (Larse n 2000; Schoeninger and DeNiro 1982; Schoeninger and Schurr, 1994; van der Merwe 1982). An additional type, Crassulacean Acid Metabolism (CAM), occurs predominantly in non-terrestrial plants and is rarely consumed by humans, the ex ception of such being seaweed and cactus (Boutton 1991b). Carbon isotopes are strong ly fractionated when plants metabolize carbon dioxide during these type s of photosynthesis and plants consequently demonstrate either high proportions of 13C or 12C (DeNiro and Schoeninger 1983; Dupras 2001; Hoefs 1980). C3 plants are typically found in modera te climates and include Old World domesticates such as barley, wheat, and rye, as well as fruits, tubers, and various shrubs and trees (Figure 3.3). C4 and CAM plants such as maize, sorghum, millet, and other cereal grasses are often found in tropical areas. C3 plants use the enzyme RuBP

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48 carboxylase, which discriminates against 13CO2, during photosynthesis and are thus known to have lower carbon st able isotope ratios than C4 and CAM plants, that use the enzyme PEP carboxylase (Boutton 1991b). C4 and CAM plants are more efficient at conserving water as their photosynthesis is performed at night, and thus thrive in increased temperatures and/or desertic environments (Sm ith and Epstein 1971; OLeary 1988). The warm, arid climates encounter ed throughout South America have yielded exceptionally suitable environments for maize agriculture. Stable carbon isotope ra tios are reported as 13C values in (read, parts per mil), relative to an accepted reference standard, and are derived from the following formula: 13C = [{(13C/12C)sample / (13C/12C)PDB} 1] x 1000 The Pee Dee Belemnite (PDB) carbonate is the commonly agreed reference standard for 13C measurements; it is derived from a piece of Cretaceous marine fossil from the Pee Dee formation in South Carolina (Larsen 2000; Schoeninger 1985; van der Merwe 1982). The National Bureau of Standards now relates current standards as the original limestone has been completely expended. C3 plants have lower 13C values (approximately ) than C4 plants (approximately ). CAM pl ants range from to but most commonly yield values of to Schoe ninger and Schurr (1994 ) postulate that a hunter-gatherer (pre-maize) diet would be represented by a human bone collagen value of approximately a 50 percent mai ze diet would be represented by a 13C value of -14 and a 13C value of -7 would indicate a di et consisting only of maize (see Figure 3.3), although nutritionall y unfeasible. It is also important to note that atmospheric CO2 has decreased from deforestation and combustion of fossil fuels by

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49 about 1.3 to 1.5 since the Industrial Revoluti on of the 1800s (Boutton 1999b; Friedl et al. 1986). Thus, individuals that lived prior to the industrial era will demonstrate slightly enriched values if evaluated by co mparable modern day samples. Nitrogen Isotope Analysis The analysis of stable isotopes of nitrogen (14N and 15N) by mass spectrometry has proven particularly useful for comparing terrestrial versus marine-based diets, as nitrogen is another essential element for plan ts. While most terrestrial plants absorb nitrogen that is deposited in the soil by decaying vegetation, marine plants absorb their nitrogen directly from the air; this produces di fferent ratios of stable nitrogen isotopes in each type of plant (Larsen 2000; Mulvaney 1993). Nitrogen isotope ratios are reported as 15N values relative to the standard of atmospheric nitrogen (N2) known as ambient inhalable rese rvoir (AIR), and are derived from the following formula: 15N = {[(15N/14N)sample (15N/14N)AIR] / (15N/14N)AIR} x 1000 Marine plants are known to have higher stab le nitrogen isotope ratios than terrestrial plants, by approximately 4 (Schwarcz and Scho eninger 1991). These values are passed through the trophic levels, with marine an imals also expected to have higher 15N values than terrestrial animals due to the type of plants they consum e (Schoeninger et al. 1983). Schoeninger et al. (1983) note th at humans on marine-based diets display bone collagen values of approximately 17 to 20, while 15N values of 6 to 12 indicate terrestrial diets.

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50 In prehistoric coastal settings, humans ar e expected to have high stable nitrogen isotope ratios due to a diet based on marine-resources (Mul vaney 1993; Schoeninger 1985). Low nitrogen ratios were expected in this study due to location of the sites in question. Although Argentina has historically been linked to Chile and Peru through exchange, the time period is too early to e xpect a high degree of marine resources in human diet. Figure 3.3. Carbon isotope fractionation in terrestrial plants. From Tykot (2004: 435.)

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51 Strontium Isotope Analysis The three types of isotopic studies most commonly unde rtaken in archaeological analysis employ carbon, nitrog en, and strontium elements. Strontium isotope ratios (87Sr/86Sr) portray the local geology of an i ndividual, by representing the strontium isotope levels of the soils of ones home range or catchment, that have been absorbed by the plants in the local ecology (S chwarcz and Schoeninger 1991). Proportions of 87Sr and 86Sr can be valuable when studying migration patterns, as different strontium isotope levels in human tooth enamel may indicate a resettlement during the individuals li fetime, particularly if the indivi dual were to move to and from heterogeneous geological areas (Ericson 1989; Schoeninger 1985). Analyzing tooth enamel, which is formed during various periods as a juvenile, from multiple permanent teeth of the same individual has the potential to identify reset tlement to a detailed annual age (Ericson 1985). Problems with strontium isotope analysis are outlined by Ericson (1989). They include: geological variability, home range de finition, and home range characterization. The documentation of migration patterns requires that an individual move to and from noticeably heterogeneous areas and that they are not extremely mobile, so as not to render chronologies extremely perplexing. Due to the difficulties associated with this type of destructive analysis, samples were not used to analyze strontium isotope ratio variation; thus strontium studies that are typi cally used to characterize migration patterns will not be further discussed in this dietary analysis. Further studies may explore this topic if a high degree of relocation be comes suspected of the subject population.

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52 Sources of Error Isotopic ratios may be mistak enly interpreted if sources of error are not taken into account. It is thus important to note factors that should be taken into consideration, as well as research that demonstrates isotope ra tios do not naturally vary by either sex or age. DeNiro and Schoeninger (1983) outline three key factors that are important to consider when reconstructing the diet of a population: (1) the isot opic composition of an individual does not significan tly differ from the mean value of the remainder of the population; (2) isotopic ratios from different bones of the same individual are virtually analogous with variation of only about 1 be tween multiple bones; and (3) isotopic ratios of males and females fed the same diet are identical. Additionally, Tieszen and Fagre (1993) and Schoen inger and Schurr (1994) advise of potential inconsistencies of 13C values when using multiple samples from a broad temporal range; these discrepancies can be caused by anthropogenic addition of CO2 into the modern day atmosphere, environm ental fluctuations, and the absorption of water. They recommend that floral remains f ound in association at a site are analyzed as well as the human samples themselves, whenev er possible. All practical matters have been taken into consideration when comparing the 13C values of associated floral remains to those of biological materials. This study also analyzes floral remains, found in association with the above-mentioned individual s, to establish the ex tent of variation in isotopic levels. Issues of interpretation can be further complicated if a lack of osteological materials limits sample size. Thus, when choosing individuals to be statistically representative of their resp ective populations, samples ha ve been chosen that are

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53 expected to be homogenous to their total group. Isotopic ratios have been tested from as many materials as possible to ensure equal va lue, and both males and females have been tested, when available, to evaluate whether they are feeding on the same diet with equal isotopic values, from the largest number of i ndividuals possible. Se asonal variation in the southern hemisphere has been taken into account, with 13C values expected to be approximately six months out of phase with the northern hemisphere (Boutton 1991b). Perhaps the most probable contaminants of stable isotope results are biological inclusions, such as another bone matrix, fa tty oils commonly calle d lipids, and humics residual components of decaying plant ma tter (White and Schwarcz 1994). Proper procedures for the removal of such matter are discussed in the followi ng chapter, as well as the various methods that were used to process the samples.

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54 Chapter Four: Stable Isotope Analysis Methods The materials supplied by the Museo de Hi storia Natural, Argentina provide an excellent opportunity for stable isotope anal ysis as they were obtained from a desert environment and therefore exhibit high le vels of preservation. The mummified soft tissue remains of the individuals in this study allow analysis of seasonal variation, migration patterns, and differing metabolic processes, with minimally destructive techniques (Fenner 2002; OConnell and Hedges 1999a, b; OConnell et al. 2001). Samples available include human bone, teeth, mu scle and skin tissue, and hair. There has been little documented analysis of soft ti ssue materials and, while this research is not entirely experimental, it will add to scie ntific knowledge by allowing examination of areas that are currently underrepresented in the literature. Objectives for this study included defining a catalogue of prehistoric stable isotope values for the Argentine provinces of San Juan and Mendoza, documenti ng any dietary transitions including those from foraging to agriculture, and examining the possible existence of separate dietary practices in the two provinces. Procedures were developed (Tykot 2002) to prepare samples for mass spectrometry and separate the complexes of mate rials described in Chapter Three. Proper execution of these techniques is necessary to ensure accurate results. Without such methods, isotopic samples could be affect ed by contaminants like residues of decomposed plants, called humates or humic acids, bacterial residues, or other similar biological complexes. For example, an intended sample of bone apatite would demonstrate inaccurate values if poorly preserve d or if another biological material that portrays different dietary rati os is included in the fina l product for analysis. The

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55 procedures outlined in this chapter are believed to be the best possible methods to remove such impurities and obtain accurate results. Sample Preparation The following pages detail the sample prep aration that was performed at the USF Laboratory for Archaeological Science in the Fall 2003 and Spring 2004 semesters, where there exist appropriate f acilities for stable isotope analysis and an experienced faculty member, Dr. Robert H. Tykot, knowle dgeable of dietary studies in the study region. Separate methods were required for each of the biological complexes discussed in the previous chapter before samples were transported to the USF Marine Science Laboratory in St. Petersburg for mass spectrometry. Bone apatite Bone apatite is typically the first complex obtained, because the collagen preparation process is minimally destructive and removes the entire mineralized portion of bone. To begin, bones that had been previ ously treated for pres ervation required an extra step of processing. Those samples were soaked in acetone and rinsed with distilled water to eliminate any conservation chemical s. Once those materials were presumably removed, segments of bone weighing approximately 1 g were cut from all samples using a circular saw. These portions were then cleaned manually and ultrasonically to remove any loose particles of soil. Distilled water was always used in the cleaning process to prevent possible contamination of any a dditional organics. Whenever necessary,

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56 ultrasonic cleaning was repeated until water ran clear. The clean bone was then dried a minimum of 6 hours (h) or overnight in a dr ying oven set at 60C. Dried samples were digitally photographed and any a bnormalities were recorded. Next, approximately 10 mg of powder were drilled from the clean, dry bone samples. Bone powder was weighed on a Mettler Toledo AT261 DeltaRange scale and the initial weight was recorded on the USF Bone Apatite Sample Processing form (Appendix C) and in the Laboratory for Archaeol ogical Science database that also assigns laboratory numbers to samples. Bone powder was transferre d into a 1.5 ml centrifuge tube labeled with the USF number and any additional material was stored in case further analysis becomes necessary. Extra portions of w hole bone that were not drilled were stored in containers that were labeled with corresponding collage n analysis numbers, with the intent of minimal destruction. Then, 1 ml of 2 percen t bleach solution was added to all apatite samples to remove collagen, bact erial proteins, and humates. The bleach solution was removed with a pi pette and replaced by distilled water after a 72 h soaking period. This rinsing pr ocedure was repeated four times before removing the water and placing the bone powde r in the drying oven for another six hour to overnight period. Once dry, samples were measured again and weights were recorded on apatite processing sheets. Apatite samples were next treated with 1 ml of buffered acetic acid solution for 24 h. This process removes non-biogenic carbonate s. Again, samples were centrifuged to pour off acetic acid and replaced with distille d water four times before placing in the drying oven. Finally, dry samples were removed from the oven, weighed to 1 mg

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57 samples and transferred into containers for transport to the mass spectrometer lab. Any excess bone powder was saved, placed in a steril e container, labeled as processed apatite, and is stored in the USF lab. Tooth enamel The outer portion of a human tooth, ca lled enamel, is composed of the same essential minerals as bone ap atite and thus requires sim ilar processing methods. Both complexes can be reduced to the general formula (Ambrose 1990): Ca5[PO4] 3OH. Enamel is much harder than apatite because it contains less organic material; in fact, it is the hardest substance in the human body. The only difference in laboratory procedures of tooth enamel (Appendix D) and bone apatite is that enamel powder does not need to remain in the bleach solution for as long a pe riod due to the reduced portion of organics to be removed and any contaminants. After cautiously drilling (Figure 4.1) for enamel and avoiding any dentin, the powder is soaked for only 24 h to remove bacterial proteins and humates, rather than the 72 h required for apatite powder. Bone collagen Although collagen typically represents an approximate 20 25 percent of fresh bone weight and is relatively insoluble, pr eservation conditions can greatly affect the amount of collagen that remains in preserve d bone. Thus, laboratory procedures most often emphasize both maximum potential yiel ds of samples and the removal of all possible contaminants (White 1991). The mo st common impurities that might affect

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58 collagen samples are other bone complexes (e.g ., apatite), lipids, and humic inclusions. Collagen samples were prepared after fi rst removing apatite samples through the procedure described above. Once apatite sa mpling was complete, the remaining 1 g of bone was processed to remove as many of th ese other materials as possible while still maintaining a relatively large portion of the collagen itself. Laboratory procedures (Appendix E) bega n by soaking each sample in 50 ml of a solution of 0.1 M sodium hydroxide (NaOH) fo r a period of 24 h. This process removes humic acids before all samples are rinsing thoroughly in distilled water (Figure 4.2). Then, 50 ml of 2 percent 2 M hydrochloric aci d (HCl) was added to remove the mineral portion of bone. This solution must be repl aced every 24 h for at least three days to remove all remaining hydroxyapatite. When the bone was completely dimineralized, the HCl was removed and the bone was thoroughly rinsed. Another cycle of NaOH for 24 h was used to ensure the elimination of any residual humic inclusions. Figure 4.1. Drilling for tooth enamel sample

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59 The fat content or lipids of the bone were removed with a defatting solution consisting of a mixture of 2 parts methanol, 1 part chloroform, and .8 parts of distilled water. This solution was allowed to soak fo r 24 h before being disposed of in special waste containers. Finally, the remaining coll agen was rinsed extra thoroughly, cut into small portions, and transferred into two-dram vials labeled with indelible marker. Samples were dried overnight before collagen yields were calculat ed and 1 mg portions were weighed and transferred into alumin um containers for transport to the mass spectrometer lab. As with the apatite sample s, any excess collagen was saved, placed in sterile containers, labeled as processed collagen, and is stor ed in the USF Laboratory for Archaeological Science. Figure 4.2. Collagen samples and the chemi cal solutions used for processing

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60 Resource samples Thirty-eight prehistoric samples of both floral and faunal resource species were analyzed as well as control samples (Figure 4.3) in an attempt to rule out the possibility of any abnormal carbon or nitrogen results that might affect human values irregularly. Table 2.2 of Chapter Two lists the types of sample s that were analyzed to this effect. All of the species named have been recovered through archaeological investigation and are thought to have played st rong roles in the diets of native Argentineans. The bones of animals such as prehistoric armadillos, Andean geese, and guanacos were prepared in the same manner as human samples; both collagen and apatite samples were taken from these bones whenever possible. Thus, the procedures for non-human animal bones are the same as those listed in Appendices D and E that were originally developed for human bones. Botanical samp les consisting of plants such as maize, winter squash, and the common bean were crushed into a powdered form using a mortar and pestle. All samples were then weighed in to appropriate sample sizes and sent to the St. Petersburg laboratory. Hair Analysis of scalp hair and skin sample s is relatively uncommon due to intrinsic preservation issues, with a few notable st udies having been performed by researchers such as Schwarcz and White (2004) and Mack o et al. (1999a). The preliminary study included in this thesis is meant to addre ss issues of preparati on and interpretation. Samples consist of eight individuals from a wide temporal span of 4070 to 590 BP, representing a number of cu ltural periods of the Arge ntine Cuyo (see Table 2.3).

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61 Admittedly, this short study requires much more work and many more samples from different time periods before any conc lusions can be made. Although this portion of the study concentrates primarily on met hods, motivating insights were provided by the work done thus far and will be analyzed in the following chapter. Procedures to prepare hair samples (A ppendix F) for mass spectrometry followed those of Fenner (2002), after OConnell and Hedges (1999a). It is particularly important to wear either a hair net or a ball cap when processing hair samples to prevent possible contamination by the modern-day researchers own hair segments. Accordingly, proper laboratory gear was worn during all pro cedures. To begin, a hair sample of approximately 15 strands was selected from one individual. Locks of hair were carefully lined up and cut along the shaft into 2 cm samples representing two month growth Figure 4.3. Archaeological samples for stable is otope analysis. Clockwise from upper left: muscle tissue; winter squash; and human mandible with teeth in situ for tooth enamel, bone apatite and collagen analysis.

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62 periods, or approximately 57 days, based on a growth rate of 0-35 mm per day (White 1993). It is noted that the time periods repr esented by hair samples are not months in a strict calendar sense (e.g., Ja nuary through February); rather the term month is used to denote an approximate growth period of 28 days. While some individuals (Table 4.1) had as few as three samples available (repres enting six months), others had as many as eight (or 16 months). Appropriate cleaning proce dures were performed on the Argentine samples to minimize the presence of any possible contam ination factors, including body fats, soil residues, or even scalp lice. Hair is almost entirely compos ed of the protein keratin, and thus requires very little pro cessing to prepare. Some researchers have even chosen to simply clean samples ultrasonically and then move on to mass spectrometry. However, amino acids residing in endogenous lipids have the potential to affect isotopic values unpredictably (OConnell and Hedges 1999a). Thus, it was decided prudent to employ a brief lipid extraction. This short procedure takes less than one day, plus drying time, and it may allow for greater reliability in the results although others, notably White (1991), have shown no differences in isotopic values Nevertheless, hair is very valuable archaeologically and, because the process wa s largely unlikely to cause any unintended harm to the samples, the lipid extraction wa s carried out to remove any variables that could not be accounted for otherwise. Each 2 cm was placed in a test tube with a sample identifier prior to execution of the lipid extraction. The test tubes were then filled with distilled water and samples were cleaned ultrasonically. Next, the water was ex changed for a 2:1 solution of methanol and chloroform, similar to the defatting solution used for collagen samples. Hair samples

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63 Table 4.1. Number of hair samples per individual Museo Sample # USF Sample Series #s Total # of Samples Total # of "months" represented SJ2 7105.1 7105.4 4 8 SJ3 ENT3 7109.1 7109.5 5 10 SJ5 ENT2 7145.1 7145.5 5 10 SJ6 ENT8 7146.1 7146.5 5 10 SJ7 ENT2 7148.1 7148.3 3 6 SJ8 ENT5 7153.1 7153.5 5 8* SJ9 ENT1 7156.1 7156.8 8 16 SJ10 ENT1 7157.1 7157.8 8 16 *Results not available for one sample due to a mechanical error in mass spec analysis were sonicated in this solution for 15 min, be fore the solution was refreshed and allowed to sonicate for another 15 min. The samples we re then rinsed with distilled water before being placed in a drying oven set to 60C to dry for 48 h. Afterward, hairs were examined microscopi cally to confirm cleanliness. Samples were ultimately weighed (Figure 4.4), folded into tin foil squares (Figure 4.5), and sent for mass spectrometric analysis. Unlike th e bone collagen tests, where two samples are taken from one individual and both run to determin e reliability, hair is unlikely to have as much trophic level variation w ithin one area as any skeletal element, with only a 1-2 difference expected between 13C values of hair samples a nd dietary proteins (OConnell and Hedges 1999a; White 1993). Also, there is often a limited amount of material, so it was deemed necessary to run only one sample for each period of two months. Results discussed in the subsequent chapter confirm that correspond ing values were dependable.

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64 Figure 4.5. Hair samples are placed in tin foil squares Figure 4.4. A hair sample representing a two-month growth period

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65 Skin and muscle Stable isotope analysis of skin and musc le tissue analysis is relatively uncommon, not only due to preservation issues, but also because of difficulties including lack of established processing procedure; ambiguity in separating complexes as was discussed in Chapter Three; and complexity in interpre ting results, which can be escalated again by uncertainty in skin or muscle sample type. This study used techniques based on the work of Schwarcz and Schoeninger (1991) to pr ocess soft tissue samples before mass spectrometric analysis. Skin and muscle samples were first clean ed ultrasonically with distilled water, and examined macro and microscopically to de termine their context. Please note that while the researchers who originally sample d the mummifed tissues believe they have appropriately differentiated between skin and muscle tissue samples, the depth of skin can be quite challenging to determine in mu mmified individuals a nd it is possible to confuse the two types when obtaining samples. Thus, all samples were photographed and assigned to categories of skin or muscle (Table 4.2) but, because it was not entirely clear what lipids and amino acids would be included in the tissues based on type, a short lipid extraction was performed on all samples. Tissues were soaked in a 2:1 methanol/chloroform defatting solution for 1 h, before being thoroughly rinsed, and placed in the drying oven overnight. A homogenized sample was considered most desirable given the conditions of the samples. Different levels of skin and mu scle depth represent various dietary periods because soft tissues constantly regenerate. T hus, it is very problematic to analyze diet if the exact depth of the tissues is not known. Add itionally, there are many factors that may

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66 further affect width, including di fferences in tissue growth, loss of external skin layers in burial, and natural decompositi on. Therefore, tissue samples were ground into a fine powder before being analyzed. The dried soft tissues proved to be very tough and fibrous, particularly after the lipid extraction was performed. Archaeologica lly valuable sample could have easily been lost if ground into the necessa ry powdered form with the typical mortar and pestle. To combat this, samples were frozen with liquid nitrogen. This converted the samples into very brittle, yet durable fro zen states, allowing them to be ground quickly before being weighed and placed into tin cups for analysis. Finally, all samples were transported to USF Tampa Marine Sciences in St. Petersburg for mass spectrometric analysis. Table 4.2. Skin and muscle sample types Museum Sample # USF Skin #USF Muscle # SJ1-ENT7 7103 SJ2 7107 SJ3-ENT3 7383 SJ4-ENT2 7141 SJ5-ENT2 7144 SJ6-ENT8 7147 SJ7-ENT2 7149 SJ8-ENT5 7154 SJ9-ENT1 7155 SJ10-ENT1 7158

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67 Instrumentation Collagen and other organic samples were processed using a CHN analyzer in combination with a Finnigan MAT stable is otope ratio mass spectrometer. The CHN analyzer switches between gases of carbon diox ide and nitrogen while samples are on the way to the mass spectrometer. This allows for samples to be mechanically transferred into the machine in ordered rows and efficiently processed. The mass spectrometer then combusts the samples and measures them in comparison to the gas standards discussed in Chapter Three. Proper operation of the mach inery is necessary to ensure precision, and values of carbon to nitrogen are analyzed after processing to confirm inte grity of results. Apatite samples were processed using an automated Kiel III individual acid bath device connected to anothe r Finnigan MAT mass spectrometer. The Kiel system sequentially drops 90C phosphoric acid into eac h sample vial. The calcium carbonate in bone apatite reacts and becomes carbon dioxide which is then measured in the mass spectrometer against the PDB standard. Re sults around 0.1 precision are typically given by both mass spectrometers (Tykot 2004).

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68 Chapter Five: Results This chapter explores the results of the stable isotope analyses. Very well preserved human and non-human animal rema ins ranging in age from approximately 4100 to 200 BP were provided by the Museo de Hi storia Natural. A total of 198 samples from 45 human individuals recovered from th e provinces of San Juan and Mendoza were analyzed to examine dietary patterns in th e prehistoric Argentine Cuyo. Materials consisted of bone collagen and apatite, toot h enamel, hair, and skin/muscle tissue. Human samples were analyzed, as well as 15 different species of potential food resources, such as llamas, rheas, maize, and squash, with numerous materials tested for all. Archaeologically, the Cuyo is thought to represent a boundary between the hunter-gatherers of Patagonia and the sedentary populations of the Andes. While San Juan is known as an area of moderate agricultural pr oduction, Mendoza is thought to have been one of the last South American culture areas to adopt maize agriculture, probably around 2000 BP (Gil 2003). This chap ter addresses the questions previously stated in Chapter Two by attempting to document whether an agricultural transition occurred, when it may have happened, and whether there were dietary differences between the two provinces in prehistoric times. Quantitative analysis was performed us ing SPSS 13.0 software for Windows at the USF Laboratory for Archaeological Science. Graphical elements, such as tables and charts, were composed using both SPSS 13. 0 and Microsoft Office Excel 2003. All mathematical operations were run at least twice to protect against any computational error. Complete results of the stable is otope analyses are provided in Appendix B.

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69 Resource Sample Results It is important to consider the results obtained from the dietary resource samples that were analyzed before examining the values provided by the analysis of human samples. While the main focus of this thesis is human values, a database of resource values is also being built. Stable isotope analysis operate s on the basic principal that varying isotopic values of hu man samples reflect different dietary patterns. However, this is complicated by the effects of trophic level: for example, a human who eats animals that have high 13C and/or 15N values, will have higher isotope values than a human who only consumes animals that have low isotope values. Thus, isotopic values could be misinterpreted if high 13C and 15N values of dietary resources are not taken into account before further analysis. Prehistoric samples of both floral and fa unal resource species were analyzed in an attempt to rule out the possi bility of any abnormal carbon or nitrogen ratios that might affect human values. The results of thes e analyses are reporte d in Table 5.1 and illustrated in Figure 5.1. When this informati on is compared to the expected values given in Figure 3.3, it is s hown that the very low 13C values of floral samples tested is consistent with standard values for C3 plants. These results suggest that all wild herbivorous animal diet s were based only on C3 plants. Thus, it is assumed that the dietary resources obtained from the Argentin e Cuyo compose a reasonably valid dataset, without abnormal resource values affecting human values irregularly. Moreover, where two or more samples from the same species ha ve been analyzed, they have resulted in similar values, particularly for 13C. The majority of species variation is seen in 15N values. This variation could

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70 Table 5.1. Results of analysis of floral and faunal resource samples Museum Sample # USF Lab # 13C 15NSpecies Material R-A-1 5905-10.7 Lama guanicoe Apatite R-A-1 6170-19.04.3 Lama guanicoe Collagen R-A-1 7368-14.29.0 Lama guanicoe Collagen R-A-2 5906-6.8 Lama guanicoe Apatite R-A-2 6171-14.75.0 Lama guanicoe Collagen R-A-2 7369-18.36.7 Rhea americana Collagen R-A-3 5907-11.1 Lama guanicoe Apatite R-A-3 6172-19.44.6 Lama guanicoe Collagen R-A-3 7370-18.15.6 Lama guanicoe Collagen R-A-4 6173-9.1 Lama guanicoe Enamel R-A-5 5908-11.5 Cholephaga melanoptera Apatite R-A-5 6174-22.04.1 Cholephaga melanoptera Collagen R-A-6 5909-11.8 Rhea americana Apatite R-A-6 6175-20.05.7 Rhea americana Collagen R-A-7 5910-12.1 Pterocnemia pennata Apatite R-A-7 6176-20.64.6 Pterocnemia pennata Collagen R-A-8 5911-9.1 Lagidium viscacia Apatite R-A-8 6177-19.33.7 Lagidium viscacia Collagen R-A-9 5912-11.1 Chaetophractus villosus Apatite R-A-9 6178-17.75.6 Chaetophractus villosus Collagen R-A-10 5913-8.9 Lama guanicoe Apatite R-A-10 6179-18.84.3 Lama guanicoe Collagen R-A-11 5914-11.5 Pterocnemia pennata Apatite R-A-11 6180-21.04.9 Pterocnemia pennata Collagen R-V-1 6181-9.73.4 Zea mays Plant R-V-2 6182-9.63.9 Zea mays Plant R-V-3 6183-23.213.1 Cucurbita maxima Plant R-V-4 6184-25.4 Lagenaria sp. Plant R-V-5 6185-27.66.9 Chenopodium sp. Plant R-V-6 7376-24.37.0 Cucurbita maxima Plant R-V-6 6186-23.9 Prosopis sp. Plant R-V-7 6187-25.41.6 Cassia arnottiana Plant R-V-8 6188-24.05.5 Phaseolus vulgaris Plant R-V-9 7379-24.29.8 Cucurbita maxima Plant R-V-9 6189-20.214.0 Geoffroea decorticans Plant R-V-10 6190-20.8 Geoffroea decorticans Plant R-V-11 6191-24.911.6 Prosopis sp. Plant R-V-13 6193-24.41.6 Schinus polygamus Plant

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71 reflect the high elevation of th e area, as nitrogen varies naturally by altitude, rather than just diet (DeNiro and Schoeni nger 1983). Although this is not an issue for many stable isotope studies, the mountainous terrain of the Andes may have affect ed these values. The domesticated llama, however, must ha ve consumed a significant amount of C4 plants, presumably cultivated maize, given their relatively high 13C values. This is in keeping with hypotheses by Gil (2003) and P earsall (1992) that humans used maize to feed their livestock, potentially before consuming the maize themselves, and should be kept in mind when investigating the isot opic values that are provided by the human

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72 samples of the Argentine Cuyo. Thus, high 13C values in human samples may be an indicator of either a diet based directly on maize, sec ondary consumption of maize by eating animals who consumed maize, or a combination thereof. Human Hard Tissue Sample Results Next, the collagen, apatite, and tooth en amel results of all human samples from both provinces are examined. St able isotope results from bone represent the last seven to 10 years of an individuals life, whereas tooth samples represent diet at the time of tooth formation, which varies depending upon th e dentition type (Mays 1995; White 1993, 2000). Most (n=36) bone samples yielded bot h collagen and apatite results, with two producing only apatite values (Table 5.2). Th is is most likely due to the more resilient nature of bone apatite, as discussed in Chap ter Three. Tooth enamel testing was highly successful, with all processed samples (n=23) yielding isotope carbon results. Bone collagen Bone collagen is formed mostly from the protein that is consumed as part of an individuals diet (Amb rose and Norr 1993). Therefore, co llagen values largely represent changes in protein diet, of animals and/or plants. Collagen samples do, however, produce nitrogen values, whereas apatite and tooth en amel samples do not. These values can be useful in contrasting terrestria l versus marine and freshwater fish based diets, as well as trophic level. First, all samples were plotted by 13C versus 15N values (Figure 5.2) with 13C values for the entire dataset ranging from .8 to .3. It is evident that San Juan

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73 Table 5.2. 15N and 13C results from human samples 15N 13C District Individual Collagen Collagen Apatite Enamel Mendoza 12 11.4 -15.9 -11.3 Mendoza 2038 6.4 -18.8 -11.2 Mendoza AF-1082 12.9 -16.5 -11.3 Mendoza AF-1083 9.5 -16.1 -10.6 -9.8 Mendoza AF-13894 9.8 -15.0 -10.1 Mendoza AF-2000 8.9 -14.8 -7.4 -5.5 Mendoza AF-2018 11.5 -14.3 -9.8 -9.0 Mendoza AF-2019 10.4 -14.5 -10.1 -8.8 Mendoza AF-2020 11.3 -14.3 -9.5 -8.7 Mendoza AF-2021 -6.1 Mendoza AF-2022 10.5 -18.5 Mendoza AF-2025 9.5 -15.5 -8.2 -10.8 Mendoza AF-2036 9.7 -17.5 -10.1 -10.8 Mendoza AF-2072 -13.7 -7.6 -6.2 Mendoza AF-500 10.3 -13.5 -8.0 Mendoza AF-503 9.4 -13.8 -7.8 -9.9 Mendoza AF-505 11.9 -16.0 -10.1 Mendoza AF-508 10.8 -17.9 -12.1 Mendoza AF-510 10.9 -17.9 -13.0 -12.7 Mendoza AF-673 10.2 -17.2 -12.5 -12.8 Mendoza AF-681 8.7 -15.6 -10.2 -10.7 Mendoza AF-8 11.7 -17.6 Mendoza AF-828 9.8 -7.6 Mendoza AF-830 -12.0 Mendoza CS-10001 11.6 -15.7 -9.0 Mendoza ENT-2 11.7 -14.9 -10.6 -9.6 Mendoza ENT-3 7.9 -16.2 -9.2 -10.2 Mendoza GIRA-27 -11.9 Mendoza GIRA-70 -9.8 Mendoza GIRA-71 10.8 -14.0 Mendoza GIRA-831 -10.5 Mendoza JP/J4 9.8 -17.4 -13.5 -13.5 Mendoza JP-1155 10.6 -16.8 -10.2 -8.6 Mendoza JP-1352 9.9 -16.3 -10.6 -11.2 Mendoza MGA-1 10.9 -14.2 -8.9 -8.7 San Juan SJ10-ENT1 9.9 -12.3 -8.2 San Juan SJ1-ENT7 9.7 -17.3 -13.1 San Juan SJ2 9.5 -13.8 -10.1 -6.3 San Juan SJ3-ENT3 9.5 -13.3 -9.8 -9.0 San Juan SJ4-ENT2 10.1 -13.8 -10.3 San Juan SJ5-ENT2 -8.3 San Juan SJ6-ENT8 8.1 -17.3 -14.0 San Juan SJ7-ENT2 10.8 -15.3 -12.2 -9.2

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74 values tend to cluster towards the more positiv e side of the figure th an those of the entire Mendoza group which are more widely disperse d. Recall that values of .5 would indicate a diet that is entirely based on C3 plants. This suggests that San Juan diets more commonly included some maize, or animals that consumed maize, than those of Mendoza, which show a great er range of values. 15N values for the entire dataset range from 6.4 to 12.9. These values are within the terrestrial diet ra nge suggested by Schoeninger et al. (1983), and the resource samples previously discussed. Th ey also correspond with the low 15N values that were expected in this study, since the study area is blocked from access to the nearest ocean by the South American Andes. However, a w eakness of the present study is the lack of analysis of any freshwater fish samples. Given the proximity of many sites to the mountain fed rivers that r un throughout western Argentin a (Compagnucci and Vargas 1998), it is not unreasonable to think that some freshwater fish may have been consumed throughout the year. Future analyses of a quatic resource samples may lead to greater understanding of the 15N values obtained from this project. Although human 15N values are not high, they do tend to vary; previously, aquatic resources were not expected to ha ve been of great importance throughout the desert environment of the Cuyo. These findi ngs suggest it may be useful to reevaluate the relative importance of aquatic resources in sites throughout the study area. Perhaps aquatic resources were of greater importance to some prehistoric residents of Mendoza and San Juan than previously thought. This study did not test any samples of freshwater fish, although it seems that th is type of analysis would be useful in the future.

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75 -12.0 -13.0 -14.0 -15.0 -16.0 -17.0 -18.0 -19.0 13C 13.0 12.0 11.0 10.0 9.0 8.0 7.0 6.0 15N San Juan Unknown San Juan Male San Juan Female Mendoza Unknown Mendoza Male Mendoza FemaleFi g ure 5.2. 13C vs. 15N values for human bone colla g en sam p les Bone apatite Bone apatite is much less susceptible to deterioration than bone collagen, and is also useful for evaluating human diets as a w hole, rather than just the protein components that are addressed by collagen samples. A disadvantage involved when sampling this type of material is that it does not allow for 15N studies. Thus, 15N values from collagen samples have been plotted against 13C values obtained from apatite in Figure 5.3 to allow for graphical representation and compare protein versus whole diet. 13C results from apatite samples ranged from .0 to .4. On first glance, the more positive apatite 13C values may indicate subsistenc e patterns increasingly based on maize. Because apatite samples reflect the to tal diet, it should be noted that the values given should equally represent the consumption of both animals that ate maize, as well as

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76 -7.00 -8.00 -9.00 -10.00 -11.00 -12.00 -13.00 -14.00 13C 13.0 12.0 11.0 10.0 9.0 8.0 7.0 6.0 15N Figure 5.3. 13C of bone apatite vs. 15N values of bone collagen samples San Juan Unknown San Juan Male San Juan Female Mendoza Unknown Mendoza Male Mendoza Female VAR00001 direct intake of maize by humans themselves. However, ranges of 13C values for both apatite and collagen samples were similar (6.6 and 6.5, respectively). The similarity of the ranges for both apatite and collagen suggests plant foods may have dominated dietary intake. Tooth enamel Tooth enamel is very similar to apatite in that it is often well preserved in the archaeological record and composed of similar material. Stable isotope analysis of both tooth enamel and bone apatite yields only 13C values, but teeth refl ect the diet of an

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77 individual at the time of tooth formation, ra ther than the last seven to 10 years of an individuals life. Because the time period represented varies by tooth type, researchers often choose to sample only one type of tooth from thei r entire population, typi cally the third molar (M3). However, issues with availability of ma terials, such as the case of juveniles whose M3s have not yet erupted, sometimes make this unachievable. Such was the case for this study, as the sample population was selected from a museum collection that possessed mixed materials from various individuals; nevertheless, whenever possible, an M3 was used. A benefit of using tooth enamel to evaluate diet is the ability to contrast juvenile, or even pre-natal, diets with those of an adult (Dupras 2001). This could not be precisely executed to the extent of an individuals lifetime with the existing materials, but comparing the two material types allows fo r a generalized picture of childhood versus adult dietary patterns. The boxplot in Figure 5.4 shows the 13C values of apatite samples next to those of tooth enamel samples. A boxpl ot is a graphical summary of the values of a group of numbers. The upper and lower portions of the box represent the upper and lower quartiles of a variable, while the hor izontal line dividing the box represents the median of the sample. The vertical lines at the top and bottom extend to the minimum and maximum data points of the sa mple (Landau and Everett 2004). Tooth enamel samples yielded more pos itive results than respective bone apatite samples for both provinces. In particul ar, the individual, SJ2 (see Table 5.2), demonstrated 13C enamel values that were 3.8 mo re positive than their corresponding bone apatite values. These results may indicate th at diet in the earlier pa rt of a prehistoric

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78 San Juan Enamel San Juan Apatite Mendoza Enamel Mendoza Apatite Figure 5.4. 13C values of apatite and tooth enamel samples -5.00 -7.50 -10.00 -12.50 13C 32 n = 19 n = 4 n = 7 n = 31 Argentineans life included more maize than that in adulthood. Furthermore, the means of the San Juan apatite and tooth enamel datasets are different, and the difference is statistically significant at the .05 level (t = -2.503; df = 9; tcritical = 2.262; p = .034). The means of the Mendoza apatite and tooth enamel data also vary, but the difference is not statistically significant at the .05 level (t = -.748; df = 47; tcritical = 2.012; p = .458). This suggests that, while San Juan individuals consumed different diets during their juvenile and adult lives, Mendoza individuals did not.

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79 Individuals AF-2025, AF-503, and ENT3 (see Table 5.2) expressed unusual 13C apatite versus 13C enamel values. Typical tooth enamel values are slightly more positive than those of bone apatite, as trophic level has a significant effect on those teeth formed prior to weaning. However these thr ee individuals possessed more negative 13C enamel values. The lower values s uggest far less intake of C4 resources at the time of enamel formation. These individuals might be aff ecting the means of the Mendoza dataset. The disparity in values of these individuals might have been caused by differences in juvenile versus adult diets, changing di etary patterns as adults, or perhaps even migration to an agricultural area af ter the age of tooth formation (Tykot 2004). Unfortunately, the sample size, particularly for San Juan tooth en amel (n=4), was small so further analysis is certainly warranted. Human Soft Tissue Sample Results One major benefit offered by this study was the opportunity to learn more about the use of soft tissue materials for isotopic analysis. Unlike the hard tissue materials previously discussed, soft tissues have a more rapid turnover rate. Their almost constant regeneration allows a more thorough reconstr uction of diet in th e Argentine Cuyo. This thesis seeks to track dietary trans itions over a long time span, as well as short, seasonal changes using human scalp hair samples. A project performed in the fall of 2004 focused on the use of sequential segmen ts of scalp hair samples from naturally mummified individuals from prehistoric San Ju an to examine the possibility of detecting short-term dietary changes. This analysis allows one to examine the ways that humans

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80 have modified their environments, by utilizi ng a foreign crop, and th e ways these changes in the natural ecology might have, in turn, affected those populations. Table 2.1 (see page 28) displays some of the cultural changes that are thought to have taken place in the province of San Juan. This chronology was constructed using both ceramic and faunal analyses (Gil, pe rsonal communication, 2004). Over time, populations are thought to have transitioned fr om hunting and gathering, to farming, and eventually to agropastoralism before being in corporated into the In ca Empire. The hair samples come from eight individuals represen ting a wide temporal span from 4070 to 590 BP, and therefore a number of cultural pe riods of the Cuyo. Although the project concentrated primarily on laboratory me thods, the analysis also provided some interesting results. Hair samples Table 5.3 displays the 15N values obtained from the stable isotope analysis of hair samples. The first column shows the number of sequential samples, with each segment, e.g., Hair 1, representing a two-m onth period. Accounting for precision of mass spectrometric analysis, nearly all individuals de monstrate short-term dietary variation. It should be noted that less than one years wo rth of hair samples exists for many of the individuals (e.g., SJ9); thus all seasonal variation could not be included in this analysis. Cultural factors, such as a spring growing season or summer harvest, might not be represented. Individuals are arranged in an increasing chrono logical order from left to right. Nitrogen values, which may indicate qua ntity of aquatic resources or mobility in altitude (Mulvaney 1993; Schoeninger et al. 1985), remain fairly consistent throughout

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81 all time periods analyzed. This is unsurpri sing due to the distance of the nearest ocean (across the Andes). This information mimi cs that obtained from the bone collagen samples, with an accepted standard offset of approximately 1 between collagen samples of hair and bone (OConnell et al. 2001), due to the re cycling of other body tissues into hair cells. Students t-tests (Table 5.4) comparing the standard deviations of 15N in the hair samples reveal that Individuals SJ8, SJ5, and SJ3 have higher variation of 15N than the analytically determined range for the dataset. Cultural factors may be viewed as a reason some members of the populati ons demonstrated higher variab ility than others. While many of these inland occupants are from sites ve ry near to river syst ems, it is not thought that the prehistoric people of the area depended heavily on fish or shellfish for their diets, with the majority of associated faunal remain s at sites consisting of terrestrial resources (Gil 1997; Gil et al. 2005). Table 5.3. 15N values for hair segments SJ7-ENT2 SJ6-ENT8 SJ8-ENT5SJ9ENT1SJ5-ENT2SJ2 SJ10-ENT1 SJ3-ENT3 4070 BP 2000 BP 2000 BP 1300 BP 880 BP 800 BP 600 BP 590 BP Hair 1 10.2 9.0 9.0 10.0 10.3 10.6 9.6 9.2 Hair 2 9.7 9.5 9.0 10.3 10.2 9.0 9.7 Hair 3 9.4 9.6 7.5 9.2 8.7 10.4 8.7 9.7 Hair 4 9.3 7.8 9.6 9.1 9.6 8.6 9.4 Hair 5 8.8 8.6 9.6 9.3 8.0 9.6 Hair 6 9.5 8.7 Hair 7 9.1 9.4 Hair 8 9.5 9.6 Mean 9.7 9.2 8.2 9.5 9.6 10.2 8.9 9.5 Range 0.8 0.8 1.6 1.0 1.6 0.9 1.6 0.5 SD 0.4 0.3 0.7 0.3 0.7 0.4 0.6 0.2 *Results not available due to a mechani cal error in mass spectrometric analysis

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82 Table 5.4. Student's t -test results for SD of 15N hair segments Individual Timea SDb t p CIlower c CIupper SJ7-ENT2 4070.40.73 .49 -.11 .21 SJ6-ENT8 2000.30 2.20 .06 -.01 .31 SJ8-ENT5 2000.70-3.67 .01 -.41 -.09 SJ9-ENT1 1300.30 2.20 .06 -.01 .31 SJ5-ENT2 880.70-3.67 .01 -.41 -.09 SJ2 800.40.73 .49 -.11 .21 SJ10-ENT1 600.60-2.20.06 -.31 .01 SJ3-ENT3 590 .20 3.67 .01 .09 .41 ayears BP bStandard Deviation (SD) cConfidence intervals (CI) represent 95 percent Note : values in bold typeface are significant at the < .05 Next, Table 5.5 shows 13C results of the analyses. The mean values are given for all hair samples tested for each individual, but do not show an overall trend towards increasing amounts of maize in th e human diet. Thus, the mean 13C values obtained from hair analysis are not in accordance with the hypothe sis that groups increasingly adopted maize agricultural practices throughout the prehistoric period. However, an examination of the standard deviations of these values, shown in the bottom row, may clarify this discrepancy. Observe the marked ly high standard deviat ions in individuals SJ8 and SJ9. Individual SJ8 is from ca. 2000 BP and associated with the period when the transition to agriculture is thought to have ta ken place. Individual SJ9 is from a period of another dietary transition, to agropastoralism. The high valu es, such as .2 for SJ8 and .5 for SJ9, could be outliers, but they may indicate actual dietary change. One-way independent sample Students t-tests (Table 5.6) of the standard deviations from the San Juan i ndividuals hair samples sugge st that Individuals SJ7 and SJ6 do not demonstrate dietary va riation as compared to the other individuals tested.

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83 Table 5.5. 13C values for hair segments SJ7-ENT2 SJ6-ENT8 SJ8-ENT5SJ9ENT1SJ5-ENT2SJ2 SJ10-ENT1 SJ3-ENT3 4070 BP 2000 BP 2000 BP 1300 BP 880 BP 800 BP600 BP 590 BP Hair 1 -16.8 -19.4 -14.2 -10.7 -15.0 -14.9 -13.4 -15.3 Hair 2 -16.7 -20.1 -12.9 -15.2 -13.4 -12.7 -16.2 Hair 3 -17.3 -20.6 -19.4 -13.4 -15.5 -13.5 -13.6 -16.9 Hair 4 -20.8 -20.0 -16.5 -15.2 -14.4 -13.2 -16.6 Hair 5 -20.2 -19.8 -10.6 -17.1 -13.9 -15.6 Hair 6 -11.0 -15.1 Hair 7 -10.3 -16.6 Hair 8 -10.0 -15.6 Mean -16.9 -20.2 -18.4 -11.9 -15.6 -14.1 -14.3 -16.1 Range 0.6 1.4 5.8 6.5 2.1 1.5 3.9 1.6 SD 0.3 0.6 2.8 2.2 0.9 0.7 1.4 0.7 *Results not available due to a mechani cal error in mass spectrometric analysis However, Individuals SJ8 and SJ9 demonstrat e greater standard deviations than the established range of variation for this dataset. Thus, it co uld be argued that heightened dietary variability began around 2000 BP and decreased after 1300 BP. Furthermore, it seems there was variab ility within diets of the San Juan population during the time period of 2000 BP. Individuals SJ6 and SJ8 are both from the same time period and, while the results of the Students-t indicate high variability for Individual SJ8, Individual SJ 6 does not show as much dietar y variation. This difference might be attributed to cultural factors; again, 2000 BP has been suggested as the time at which agriculture began in San Juan (Gil 2003) Perhaps some individuals were still eating highly varied diets, while others bega n to increasingly cons ume maize products at the expense of other foods. This information may indicate that the times before, during, or after a dietary a transition were particularly inconsistent, with seasonal patterns eventually steadying out over long periods.

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84 Table 5.6. Student's t -test results for SD of 13C hair segments Individual Timea SDb t p CIlower c CIupper SJ7-ENT2 4070.302.91 .02 .17 1.63 SJ6-ENT8 2000.60 1.94 .09 -.13 1.33 SJ8-ENT5 20002.80-5.17 .00 -2.33 -.87 SJ9-ENT1 13002.20-3.23 .01 -1.73 -.27 SJ5-ENT2 880.90.97 .37 -.43 1.03 SJ2 800.701.62 .15 -.23 1.23 SJ10-ENT1 6001.40-.65 .54 -.93 .53 SJ3-ENT3 590 .70 1.62 .15 -.23 1.23 ayears BP bStandard Deviation (SD) cConfidence intervals (CI) represent 95 percent Note : values in bold typeface are significant at the < .05 The information given here suggests that diets during times of transition were more irregular, based upon the Students t-test of the standard de viations of sequential hair samples. These results can be explaine d in a variety of ways. For example, some researchers (e.g., Price and Gebauer 1995) argue that popula tions consciously began to produce food in satisfactory ecological systems as a preemptive way of dissolving potential risks of dietary stre ss in future times of need. Alternatively, authors such as Gilber t (1985) and Keene ( 1985) suggest that environmental or nutritional deficiencies necessitated the adoption of alternative means of food acquisition. Some anthropologist s (e.g., Larsen 1995, 2000; Smith et al. 1984) point to detrimental changes in health that correspond to th e adoption of maize agriculture, such as anemia and osteoporosis. It has even been not ed (Steckel et al. 2002a) that agricultural so cieties were the first to fall vi ctim to the health epidemics brought on by the invasion of the Spanish. This might have been due to weakened immune systems brought on by inadequate diets, thereby making groups such as the Inca

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85 easy prey for European disease vectors. Fu rther, some members of the Argentine study population demonstrated existence of nutritional stress indicators, such as dental caries, that can be viewed in relation to the dietary shifts that may have been a cultural adaptation to the envi ronment of the Cuyo. Admittedly, the small number of samples analyzed in this portion of the study requires much more work before any firm declaration can be made about dietary transitions and seasonal varia tion. It would be particul arly worthwhile to analyze individuals from the periods surroun ding 2000 and 1300 BP to document whether heightened variation actually occurred, and when, if at all, it began and eventually decreased. This method of analysis c ould address many of th e current hypotheses regarding the motivation for the transition to agriculture. Osteological studies would be useful to examine the effects of dietar y transitions and to document any physical evidence of malnutrition. Additional testing of soft tissue samples has the potential to shed light on these topics. Skin/Muscle tissue Sampling for skin and muscle tissues pr esented unique difficulties including uncertainty of tissue type, depth of tissue sample, and variable preservation levels. Nevertheless, procedures were developed to prepare samples as best as possible under the given circumstances; the primary benefit of this portion of the study was intended to be the assessment of laboratory procedures. Li pids were removed us ing a defatting solution and samples were frozen using liquid nitr ogen to facilitate powdering and thus homogenization of various layers of soft tissue.

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86 13C values ranged from .3 to .6 (T able 5.7). Figure 5.5 displays a boxplot comparing 13C values for hair samples (left) to 13C values for skin samples (right); outliers represent cases > 1.5 times the interquartile ra nge. This plot shows that, with the exception of Individua ls 1 and 4 for which there are no data for hair samples, the values for the hair and skin samples are generally congruent. Th e contrary cases are Individuals 5, 6, and 7, where the values fo r the skin samples are outside the range of those for the hair samples. A greater range (11.0) was given by the 15N values, with a minimum value of 11.8 and a maximum value of 22.8 (Table 5.7). Figure 5.6 compares 15N values for hair samples (left) to 15N values for skin samples (right); outliers represent cases > 1.5 times the interquartile range. This plot shows that, with the exception of Individuals 1 and 4 for which there are no data for hair sample s, the values for the skin samples are far outside the range of those for the ha ir samples displayed in Table 5.3. Short-term dietary variations could greatly affect flesh values given their rapid regeneration, but the significance of this ir regularity is not yet understood. The larger ranges of variation may indicate extremely fr equent changes in diet, or they may be regarded as erroneous due to the issues disc ussed above and in further detail in Chapter Three. Thus, it is suggested that a much la rger population size and further evaluation of laboratory procedures, to be ab le to clearly determine tissue types (e.g. skin or muscle), are necessary before any dietary inferences can be made on the basis of flesh analysis.

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87 Table 5.7. Stable isotope analysis results of skin/muscle tissues Museum Sample # USF Lab # Years BP 13C 15N Site SJ1-ENT7 7103 7900-4200 -16.4 11.8 Gruta Morrillos SJ2 7107 800 -14.4 13.5 Calingasta SJ3-ENT3 7383 590 -15.8 13.3 Punta del Barro SJ4-ENT2 7141 640 -15.3 13.1 Angualasto SJ5-ENT2 7144 880 -17.2 12.7 C Calvario SJ6-ENT8 7147 2000 -18.0 22.8 Gruta 1 Morrillos SJ7-ENT2 7149 4070 -20.3 15.0 Gruta 1 Morrillos-Morrillos SJ8-ENT5 7154 2000 -18.2 13.2 Gruta 1 MorrillosAnsilta SJ9-ENT1 7155 1400-1200 -11.6 12.3 Hilario SJ10-ENT1 7158 600 -14.1 12.0 Angualasto Mean -16.1 14.0 Range 8.7 11.0 SD 2.5 3.2 Figure 5.5. 13C values for hair (left) vs. skin (right) samples

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88 Spatial Comparison It has been previously suggested that the samples from San Juan and Mendoza represent two distinct populations that practiced differe nt food procurement strategies throughout the majority of prehistory (Gil 2003) To test this hypothesis, inter-population relationships were examined in relation to space and time dimensions to determine whether the two groups separated dietary pr actices and how these customs may have changed over time. The results were divi ded by province (Table 5.8). Means of 13C values obtained from bone collagen (-14. 7) and tooth enamel samples (-8.2) from San Juan were higher than those of Mendo za (-15.9 and -9.8, respectively). The average of bone apatite samples from Mendoza (-10.2) was slightly higher than that of San Juan (-11.1); however, the difference between the means is not statistically Figure 5.6. 15N values for hair (left) vs. skin (right) samples

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89 significant at the .05 level (t = -1.1337; df = 36; tcritical = 2.021; p = .190). The reasons for the similarities between the two groups are difficult to postulate given the relatively small sample size of San Juan samples. Additiona lly, this information includes samples from all analyzed periods, so temporal differen ces may be affecting values as well. Temporal Comparison Figure 5.7 displays 13C values of the three hard tissu e types as they vary by time. San Juan and Mendoza individuals have been grouped together to allow for a larger sample size. Bone collagen and bone apatite samples, indicative of protein portions and whole diets respectively, seem to increase throughout the long period of 6050 to 50 BP; however, ANOVA shows that dietary change wa s not significant at the .05 level for either bone collagen (f = .656; df = 20,1; fcritical = 248.013; p = .767) or bone apatite (f = 3.282; df = 21,1; fcritical = 248.310; p = .128). Additionally, values of tooth enamel samples, which represent diet during time of tooth formation, seem to have remained fairly stable during all peri ods considered. A Students t-test was used to Table 5.8. Comparison by province of human bone and tooth samples Mendoza San Juan N Mean SD N Mean SD Tissue* Bone Collagen 13C 29 -15.9 1.5 7 -14.7 2.0 15N 29 10.3 1.3 7 9.7 0.8 Bone Apatite 31 -10.2 1.6 7 -11.1 2.0 Tooth Enamel 20 -9.8 2.2 4 -8.2 1.3 *All results given are for 13C values unless otherwise stated

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90 2050-50 4050-2050 6050-4050 Years BP -5.0 -7.5 -10.0 -12.5 -15.0 -17.5 -20.0 13C Tooth Enamel Bone Apatite Bone CollagenMaterial evaluate change between the two later time periods of 4050-2050 BP and 2050-50 BP, because the dataset for the time period of 6050-4050 consisted of only one tooth enamel result. The difference between the means of the two tooth enamel datasets is not significant at the .05 level (t = -1.644; df = 17; tcritical = 1.740; p = .119). At this time, dietary change does not seem to have been great enough to significantly affect 13C values. A criticism of this anal ysis is the uneven distribution of sample sizes throughout various periods. Unfo rtunately, the sample was heavily skewed towards later individuals, with a total number of 54 bone coll agen, bone apatite, and tooth enamel 13C results from the period of 2050-50 BP. Twelve samples have been given for n = 3 Figure 5.7. 13C results for both provinces vary by period n = 2 n = 2 n = 2 n = 1 n = 7 n = 21 n = 21 n = 12

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91 the period of 4050-2050 BP, and only five from 6050 to 4050 BP. Thus, a much larger number of samples from the earlier periods should be analyzed to examine more accurately issues of temporal distribution. Discussion The analysis of floral an d faunal resource samples produced expected results, with the low 13C values of all tested C3 plants and wild herbivorous animals considered standard. Likewise, the maize samp les that were analyzed produced 13C values within the expected norm. The samples of domes ticated llama suggest that these animals consumed considerable amounts of C4 plants, most likely maize, during their lives. These results can be viewed in relation to the human samples, as the isotopic ratios of dietary resources affect the body ti ssues, including bone co llagen, bone apatite, and tooth enamel, of their consumers. 13C values obtained from both hard and soft tissue samples did not support th e hypothesis that individuals from prehistoric San Juan more commonly consumed maize, or animals that ate maize, than the residents of Mendoza. While these two geographic areas ma y differ within the archaeological record, their dietary patterns might have been more similar than previously thought. However, the small number of San Juan individuals th at were tested leaves further analysis desirable. Individuals tested from severa l Mendoza sites have average 13C values reflecting a 25-30 percent dependence on C4 products in their whole diet, as indicated by bone apatite analysis, while the 5.8 difference be tween collagen and apatite values supports the interpretation that plant f ood was of greater dietary signif icance than animal products.

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92 Considering the diverse times and contexts re presented by the indivi duals tested, the 5 range in carbon isotope values among the indivi duals tested is not unexpected and may be partly correlated with increasi ng maize dependence over time. The 15N values do not imply that large amounts of aquatic resources were consumed in either province, but the range of variation merits further investigation. 15N can vary by both altitude and dietary intake. The dramatic altitudes found in the central Andes might have affected the 15N values of resource samples and the people who consumed them. This could also be a si gnal of aquatic resource consumption or a combination of both. The analysis of freshw ater fish and additional human bone collagen samples might shed light on this issue. Most importantly, it appears that maize c ould have been a significant part of human diets in 2000 BP as hypothesized by Gil ( 2003). It is possible that the people of the prehistoric Argentine Cuyo adopted ma ize agriculture much earlier than was previously thought, and possibly as early as 6050 BP. This evidence in more in line with that of other areas of South America, such as coastal Ecuador, where Tykot and Staller (2002) used stable isotope analysis to argue that domesticated maize was an important part of human diet as early as 4000 BP. Additionally, Pearsall and colleagues (2004) have demonstrated the antiquity of maize throughout Ecuador by anal yzing residues left on stone tools from the Real Alto site. The previous hypothesis of maize diets first occurring at 2000 BP is later than the dome stication date suggested by Benz (2001) at 5400 BP. For San Juan, a much smaller number of individuals have been tested, such that, while there appears to be di fferent carbon isotope averages when compared with the

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93 Mendoza sites, the differences are not statistic ally significant. Th e hypothesis (Gil 2003) of a chronological shift toward s greater maize dependence over time is not supported by the current study, as demonstrated by the anal ysis that did not show these individuals to be statistically different from those of later time periods.

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94 Chapter Six: Conclusion The introduction of maize to prehistori c diets is of fundamental importance to understanding the evolution and development of agriculture in the New World (Tieszen and Fagre 1993). As previously stated, this introduction of a new food resource has been the subject of investigation for some time, ye t there remains a lack of information about the history of plant domestication in the s outhern Andes, where there existed Andean cultures with somewhat less emphasis on agriculture (Gil 200 3; Isbell 1997). Information on ceramic typology, lithic analys is, and settlement pa tterns dominates New World archaeology; what is known about the history of maize in Argentina has been discovered mainly through traditional archaeol ogical techniques, in the few instances when archaeologists have been fortunate e nough to come across well preserved floral and faunal remains. A lack of proper excavati on techniques, such as flotation and other systematic botanical recovery methods, may further distort some of the existing data regarding prehistoric diets (K eene 1985; Nez Regueiro 1978) The few reports that do exist have been published as a sort of gray literature, and are of ten difficult to obtain for archaeologists who are not directly invol ved in the specific project or who do not speak English (Pearsall 1992). Because South America is so geographically and culturally diverse, maize cultivation models cannot often be applied to various regions; thus, there exists a need to establish a clear chronology for th e central west portion of Ar gentina, a unique area that once existed at the edge of th e Inca Empire (Pearsall 1994). Th is disparity of information can be addressed by isotopic analysis of cons umer tissues, which provides an invaluable record of dietary change when isotopically different foods are introduced into a diet

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95 (Fogel and Tuross 2003; Schoeninger a nd Schurr 1994; Schwarcz and Schoeninger 1991). It is important to consider the cultural se tting of maize agriculture, as well as the crop itself. Differences in settlement pa tterns, material cultures, and subsistence adaptations exist among all maize-based agricultural societies (Pearsall 1994; Schoeninger and Schurr 1994). Gr emillion (1996) and Gil (2003) assert that the issue of crop adaptation can produce insi ghts into issues such as those mentioned above. This project, with the analysis of multiple tissu es, may assist other researchers working in South America who contribute to a more thorough explanation of the history of agriculture and human di etary adaptations thr oughout the New World. Considerations for Isotopic Studies Results of this study indicate that the faunal and floral re source species that were analyzed provide a suitable dataset for exam ining the use of dietary elements in the prehistoric Argentine Cuyo. While 15N values show relatively high variation when compared to corresponding 13C values, this may be linked to the dramatic altitudes of the Andes mountain range, ra ther than diet alone. The analysis of human tissues does no t support hypotheses regarding temporal and spatial variation in ce ntral western Argentina. 13C values from both provinces indicate the presence of C4 plants in human diets and while a transition from foraging to farming is indicated in the archaeological r ecord, the timing of this transition is still unclear. Further analysis ma y yield greater understanding of the dietary transition that

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96 was previously thought to have taken place around 2000 BP (Gil 2003), but now seems much earlier. While the effects of diet ary transitions are still not completely understood, the analysis of human scalp hair samples shows si gnificant seasonal variation in diet. Still, the analysis of a much larger sample of individuals is necessary before addressing hypotheses regarding the transition to agri culture. Radiographi c studies on the mummified individuals may allow for a more thorough examination of the effects of dietary transition. Scholarly and Educational Significance The combination of analyses of multiple tissues has allowed for the reconstruction of a dietary life hist ory of well-preserved individuals from the Argentine Cuyo, while advancing knowledge and expert ise in archaeological science. The results obtained affect South American archaeologists and researcher s practicing stable isotope analysis, while addressing research topics and areas that have been previously overlooked. The information given here adds knowledge to the fields of prehistoric diet reconstruction, New World archaeology, bone chemistry, and archaeological science. This knowledge can be used to enhance theories on cultura l adaptation, seasonal dietary variation, foraging versus agricultural diets and more. This project can be used to construct a preliminary chronology for the adoption of maize agriculture in the areas currently known as San Juan and Mendoza, Argentina. The results obtained add to the scientific data base of knowledge on the ability of stable isotope analysis to model prehis toric diets. The experimental soft tissue analysis adds to

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97 the understanding of laboratory procedures a nd provides essential in formation regarding seasonal variation in diet. The transition fr om foraging to farming has been linked with alterations in social organization, economic systems, politics, religion, and even disease (Larsen 2002; Schoeninger and Schurr 1994). By analysis of isotopic ratios in a quantitative manner, researchers can begin to thoroughly address these issues, and gain valuable insight into the human past. Applied Perspectives Gilbert (1985:339) notes that “the relations hip of diet to the continued survival and perpetuation of a species is fundamental to understanding that species’ interaction with its environment.” This se ntiment hints at the question of why do this type of research. The University of South Florida’ s Department of Anthropology engages in an applied approach, with an emphasis on assis ting modern day populations. The more that is learned about the beginnings of human di sease, the better the human condition can itself be understood. Diseases like anemia a nd osteoporosis may be vitally linked to the transition to agriculture. Information about the origins of these ailments used in conjunction with data obtained from stable isotope analysis can help researchers better understand, and possibly even treat or prevent, disease in populations that are struggling with these ailments today. In particular, information garnered from the soft tissue analysis portion in this study indicates dietary in stability during times of transition. Further research has been planned that may link dietary variability to weakened immune systems and disease suscepta bility prior to Spanish contact in the New World.

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98 Additionally, scientists faced with problem s of destruction and deforestation of the American rainforests are beginning to examine past food production systems (Bruhns 1994). Bray (2000) argues that archaeologi sts are in the position to make unique contributions to the topic of sustainabl e methods of tropical land use by exploring previous land management schemes and edu cating the public about both the benefits and shortcomings of such practices. Future Directions This study is but a small portion of a much larger project attemp ting to reconstruct prehistoric lifeways in the Argentine Cuyo. Researchers at the Museo de Historia, interested in dietary adaptations and socio economic systems prior to European contact, are performing ongoing archaeological examinat ions, including faunal, macrobotanical and paleoclimatic analyses. Morphometric examination of human remains and dental pathologies is in process, and lith ic and ceramic analyses are ongoing. The Laboratory for Archaeological Scien ce at the University of South Florida continues isotopic studies of osteologi cal materials, while researching funding opportunities for more detailed and extens ive analyses of human materials from Argentina.

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99 References Allison, M.J. 1984. Paleopathology in Pe ruvian and Chilean populations. In M.N. Cohen & G.J. Armelagos (eds.), Paleopathology at the Origins of Agriculture, pp. 515-529. Orlando: Academic Press. Ambrose, S.H. 1990. Preparation and ch aracterizaton of bone and tooth collagen for isotopic analysis. Journal of Archaeological Science 17: 431-451. Ambrose, S.H. & L. Norr. 1993. Experimental evidence for the relationship of the carbon isotope ratios of whole diet and dietary protein to those of bone collagen and carbonate. In J.B. Lambert & G. Grupe (eds.), Prehistoric Human Bone: Archaeology at the Molecular Level, pp. 1-37. New York: Springer-Verlag. Angel, J.L. 1984. Health as a crucial f actor in the changes fro m hunting to developed farming in the eastern Mediterranean. In M.N. Cohen & G.J. Armelagos (eds.), Paleopathology at the Origins of Agriculture, pp. 51-73. New York: Academic Press. Armelagos, G.J. 1994. "You are what you eat". In Sobolik, K.D. (ed.), Paleonutrition: the Diet and Health of Prehistoric Americans, pp. 235-244. Center for Archaeological Investigations Occasional Pape r No. 22. Board of Trustees, Southern Illinois University at Carbondale. Aufderheide, A.C., M.A. Kelley, M. Rivera, L. Gray, L.L. Tieszen, E. Iversen, H.R. Krouse, A. Carevic. 1994. Contributions of chemical dietary reconstruction to the assessment of adaptions by ancient highland immigrants (Alto Ramirez) to coastal conditions at Pisagua, North Chile. Journal of Archaeological Science 21(4): 515524. Aufderheide, A.C., I. Munoz & B. Arriaza. 1993. Seven Chinchorro mummies and the prehistory of northern Chile. American Journal of Ph ysical Anthropology 91(2): 189201. Aufderheide, A.C. & C. Rodriguez-Martin. 1998. The Cambridge Encyclopedia of Human Paleopathology. Cambridge: Cambridge University Press. Barberena, R. 2002. Los limites del mar: Isotopos estables en Patagonia meridional. Buenos Aires: Sociedad Argentina de Antropologia. Beadle, G.W. 1980. The ancestry of corn. Scientific American 242: 112-119. Benz, B.F. 2001. Archaeological eviden ce of teosinte domestication from Guil Naquitz, Oaxaca. Proceedings of the Nati onal Academy of Science 98(4): 21042106.

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100 Benz, B.F. & A. Long. 2000. Prehistoric maize evolution in the Tehuacan Valley. Current Anthropology 41(3): 459-465. Bonavia, D. & A. Grobman. 1989. Andean maize: its origins and domestication. In Harris, D.R. & G.C. Hillman (eds.), Foraging and Farming: the Evolution of Plant Exploitation, pp. 456-470. London: Unwin Hyman Ltd. Boutton, T.W. 1991a. Stable carbon isotope ratios of natural materials: I. Sample preparation and mass spectrometric anal ysis. In Coleman, D.C. (ed.), Carbon Isotope Techniques, pp. 155-171. San Diego, CA: Academic Press, Inc. Boutton, T.W. 1991b. Stable carbon isotope ra tios of natural materials: II. Atmospheric, terrestrial, marine, and freshwater envi ronments. In Coleman, D.C. (ed.), Carbon Isotope Techniques, pp. 173-185. San Diego, CA: Academic Press, Inc. Bray, G.A. & B.M. Popkin. 1998. Dietary fat does affect obesity. American Journal of Clinical Nutrition 68: 1157-1173. Bray, W. 2000. Ancient food for thought. Nature 408(9): 145-146. Brier, B. 1998. The Encyclopedia of Mummies. New York: Facts on File, Inc. Bruhns, K.O. 1994. Ancient South America. Cambridge: Cambridge University Press. Butzer, K.W. 1982. Archaeology as Human Ecology: Method and Theory for a Contextual Approach. New York: Cambridge University Press. Charters, S. 1993. Quantification and distri bution of lipids in ar chaeological ceramics: implications for sampling potsherds for orga nic residue analysis and the classification of vessel use. Archaeometry 35(2): 211-223. Clapperton, C.M. 1993. The Quaternary Geology and Geomorphology of South America. Amsterdam: Elsevier. Clark, G. 1970. Aspects of Prehistory. Berkeley: UC Press. Cockburn, A. E. & T.A. Reyman. 1998. Mummies, Disease and Ancient Cultures. Cambridge: Cambrige University Press. Cohen, M.N. 1977. The Food Crisis in Prehistory: Overpopulation and the Origins of Agriculture. New Haven: Yale University Press.

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101 Compagnucci, R.H. & W.M. Vargas. 1998. In ter-annual variability of the Cuyo rivers streamflow in the Argentine And ean mountains and ENSO events. International Journal of Climatology 18: 1593-1609. DAntoni, H.L. 1980. Los ltimos 30,000 aos en el sur de Mendoza Argentina. Coleccin Cientifica 86: 83-108. DAntoni, H.L. 1983. Pollen analysis of Gruta del Indio. Quaternary of South America and Antarctic Peninsula 1: 83-104. DeNiro, M.J. & S. Epstein. 1978. Influence of diet on the distribu tion of carbon isotopes in animals. Geochima et Cosmochima Acta 42: 495-506. DeNiro, M.J. & M.J. Schoeninger. 1983. Stable carbon and nitrogen isotopic rations of bone collagen: variations within individuals between sexes, and within populations raised on monotonous diets. Journal of Archaeological Science 10: 199-203. Dieguez, S. & G. Neme. 2003. Geochronology of Arroyo Malo 3 arch aeological site and the first human occupations in the Nordpata gonia early Holocene. In Bonnichsen, R., L. Miotti, M. Salemme & N. Flegenheimer (eds.), Ancient Evidence for Paleo South Americans: from Where the South Wind Blow s Center for the Study of the First Americans, pp. 87-92. College Station, TX : Texas A&M University Press. Dillehay, T. (ed). 1995. Tombs for the Living: Andean Mortuary Practices. Washington, D.C.: Dumbar ton Oaks Research Library and Collection. Dillehay, T. 1999. The late Pleistocene cultures of South America. Evolutionary Anthropology 7(6): 206-216. Dobres, M.-A. & J. Robb (eds.). 2000. Agency in Archaeology. London: Routledge. Dupras, T.L. 2001. Infant wean ing practices in Roman Egypt. American Journal of Physical Anthropology 115(3): 201-212. Eaton, S.B., S.B. Eaton III & L. Cordain. 2002. Evolution, diet, and health. In Ungar, P.S. & M.F. Teaford (eds.), Human Diet: Its Origin and Evolution, pp. 7-17. Westport, Connecticut: Bergin & Garvey. Eaton, S.B., M. Konner, & M. Shostak. 1988. Stone agers in the fast lane: chronic degenerative diseases in e volutionary perspective. American Journal of Medicine 84: 739-749. Elerick, D.V. 1997. Human Paleopathology and Related Subjects: An International Bibliography. San Diego: San Diego Museum of Man.

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114 Appendix A. Correspondence of Museo de Historia to USF sample numbers per individual Sample Type Museum Sample # Collagen Apatite Enamel Hair* Skin Muscle 12 7623 8198 2038 6217 6218 AF-1082 6212 6213 AF-1083 7624 8199 8196 AF-13894 7352 7353 AF-2000 7619 8191 8192 AF-2018 7354 7355 7356 AF-2019 7349 7350 7351 AF-2020 7358 7359 7357 AF-2021 7618 8189 8190 AF-2022 6194 6196 AF-2025 7333 7334 7332 AF-2036 6206 6207 6208 AF-2072 7622 8197 8194 AF-500 6222 6223 AF-503 6203 6204 6205 AF-505 6197 6198 AF-508 6209 6210 AF-510 7331 7330 7329 AF-673 7335 7336 7337 AF-681 7360 7361 7362 AF-8 7625 AF-828 7621 8195 AF-830 7620 8193 CS-10001 6199 6200 ENT-2 6226 6227 6228 ENT-3 7342 7343 7341 GIRA-27 6225 GIRA-70 6202 GIRA-71 6201 GIRA-831 7364 JP/J4 7347 7348 7346 JP-1155 6219 6220 6221 JP-1352 7340 7339 7338 MGA-1 6214 6215 6216

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115 R-V-1 (SJ) 7371 R-V-2 (SJ) 7372 R-V-3 (SJ) 7373 R-V-4 (SJ) 7374 R-V-5 (SJ) 7375 R-V-6 (SJ) 7376 R-V-7 (SJ) 7377 R-V-8 (SJ) 7378 R-V-9 (SJ) 7379 SJ1-ENT7 7381 7104 7103 SJ2 7382 7106 7108 7105.1-.4 7107 SJ3-ENT3 7111 7110 7112 7109.1-.5 7383 SJ4-ENT2 7384 7142 7141 SJ5-ENT2 7143 7145.1-.5 7144 SJ6-ENT8 7385 7380 7146.1-.5 7147 SJ7-ENT2 7152 7151 7150 7148.1-.3 7149 SJ8-ENT5 7153.1-.5 7154 SJ9-ENT1 7156.1-.8 7155 SJ10-ENT1 7386 7159 7157.1-.8 7158 *Series of numbers indicating sequential 2 centimeter samples, with .1 being closest to scalp

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Appendix B. Results of stable isotope analysis for all samples Museo Sample # USF # Material 13C 15N Species Years BP Sex Age Site 12 8198 Apatite -11.3 Homo sapiens 3850 Adult MZ; Caverna de las Brujas 2038 6217 Collagen -18.8 6.4 Homo sapiens F 35-49 yrs MZ; El Desecho 2038 6218 Apatite -11.2 Homo sapiens F 35-49 yrs MZ; El Desecho AF-1082 6212 Collagen -16.5 12.9 Homo sapiens F 35-49 yrs MZ; Agua del toro AF-1082 6213 Apatite -11.3 Homo sapiens F 35-49 yrs MZ; Agua del toro AF-1083 8196 Enamel -9.8 Homo sapiens 100 MZ; Arbolito 8 AF-1083 8199 Apatite -10.6 Homo sapiens 100 MZ; Arbolito 6 AF-13894 7352 Collagen -15.0 9.8 Homo sapiens 2300 Perinatal? MZ; Gruta del Indio AF-13894 7353 Apatite -10.1 Homo sapiens 2300 Perinatal? MZ; Gruta del Indio AF-2000 8191 Apatite -7.4 Homo sapiens 580 MZ; C Negro del Escorial AF-2000 8192 Enamel -5.5 Homo sapiens 580 MZ; C Negro del Escorial AF-2018 7354 Collagen -14. 3 11.5 Homo sapiens 1700 -1400 MZ; Canada Seca AF-2018 7355 Apatite -9.8 Homo sa piens 1700-1400 MZ; Canada Seca AF-2018 7356 Enamel -9.0 Homo sa piens 1700-1400 MZ; Canada Seca AF-2019 7349 Collagen -14. 5 10.4 Homo sapiens 1700 -1400 MZ; Canada Seca AF-2019 7350 Apatite -10.1 Homo sa piens 1700-1400 MZ; Canada Seca AF-2019 7351 Enamel -8.8 Homo sa piens 1700-1400 MZ; Canada Seca AF-2020 7357 Enamel -8.7 Homo sa piens 1700-1400 MZ; Canada Seca AF-2020 7358 Collagen -14. 3 11.3 Homo sapiens 1700 -1400 MZ; Canada Seca AF-2020 7359 Apatite -9.5 Homo sa piens 1700-1400 MZ; Canada Seca AF-2021 8190 Enamel -6.1 Homo sapiens 510 Infant MZ; Gruta del Indio AF-2022 6194 Collagen -18.5 10.5 Homo sapiens 1200 M 30-45 yrs MZ; Ojo de Agua AF-2022 6196 Enamel -11.9 Homo sapiens 1200 M 15-18 yrs MZ; Ojo de Agua AF-2025 7332 Enamel -10.8 Homo sapiens 200 MZ; Tierras Blancas AF-2025 7333 Collagen -15.5 9.5 Homo sapiens 200 MZ; Tierras Blancas 116

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AF-2025 7334 Apatite -8.2 Homo sapiens 200 MZ; Tierras Blancas AF-2036 6206 Collagen -17.5 9.7 Homo sapiens F 16-20 yrs MZ; India embarzada AF-2036 6207 Apatite -10.1 Homo sapiens F 16-20 yrs MZ; India embarzada AF-2036 6208 Enamel -10.8 Homo sapiens F 16-20 yrs MZ; India embarzada AF-2072 8194 Enamel -6.2 Homo sapiens 970 F 15-18 yrs MZ; Las Ramadas AF-2072 8197 Apatite -7.6 Homo sapiens 970 F 15-18 yrs MZ; Las Ramadas AF-500 6222 Collagen -13.5 10.3 Homo sapiens 1760 M >50 yrs MZ; Rincon del Atuel AF-500 6223 Apatite -8.1 Homo sapiens 1760 M >50 yrs MZ; Rincon del Atuel AF-503 6203 Collagen -13.8 9.4 Homo sapiens 1760 M 34-45 yrs MZ; RA-1 AF-503 6204 Apatite -7.9 Homo sapiens 1760 M 34-45 yrs MZ; RA-1 AF-503 6205 Enamel -9.9 Homo sapiens 1760 M 34-45 yrs MZ; RA-1 AF-505 6197 Collagen -16.0 11.9 Homo sapiens M 45-50 yrs MZ; La Matancilla AF-505 6198 Apatite -10.1 Homo sapiens M 45-50 yrs MZ; La Matancilla AF-508 6209 Collagen -17.9 10.8 Homo sapiens M 38-49 yrs MZ; Cerro Mesa AF-508 6210 Apatite -12.2 Homo sapiens M 38-49 yrs MZ; Cerro Mesa AF-510 7329 Enamel -12.7 Homo sapiens 300-200 MZ; Cerro Mesa AF-510 7330 Apatite -13.0 Homo sapiens 300-200 MZ; Cerro Mesa AF-510 7331 Collagen -17.9 10.9 Homo sapiens 300-200 MZ; Cerro Mesa AF-673 7335 Collagen -17.2 10.2 Homo sapiens MZ; El Manzano AF-673 7336 Apatite -12.5 Homo sapiens MZ; El Manzano AF-673 7337 Enamel -12.8 Homo sapiens Adult MZ; El Manzano AF-681 7360 Collagen -15.6 8.7 Homo sapiens 2000 MZ; Medano Puesto Diaz AF-681 7361 Apatite -10.2 Homo sapiens 2000 MZ; Medano Puesto Diaz AF-681 7362 Enamel -10.7 Homo sapiens 2000 MZ; Medano Puesto Diaz AF-828 8195 Apatite -7.6 Homo sapiens 580 F 30-49 yrs MZ; Gruta del Indio AF-830 8193 Apatite -12.0 Homo sapiens 3860 MZ; Gruta del Indio CS-10001 6199 Collagen -15.7 11.6 Homo sapiens M 30-45 yrs MZ; Canada Seca CS-10001 6200 Apatite -9 .0 Homo sapiens M 30-45 yrs MZ; Canada Seca 117

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ENT-2 6226 Collagen -14.9 11.7 Homo sapiens 400 F MZ; Capiz Alto ENT-2 6227 Apatite -10.6 Homo sapiens 400 F MZ; Capiz Alto ENT-2 6228 Enamel -9.6 Homo sapiens 400 F MZ; Capiz Alto ENT-3 7341 Enamel -10.2 Homo sapiens MZ; El Chacay ENT-3 7342 Collagen -16.2 7.9 Homo sapiens MZ; El Chacay ENT-3 7343 Apatite -9.2 Homo sapiens MZ; El Chacay GIRA-27 6225 Apatite -11.9 Homo sapiens Adult MZ; Gruta del Indio GIRA-70 6202 Apatite -9.8 Homo sapiens Adult MZ; Gruta del Indio GIRA-71 6201 Collagen -14.0 10.8 Homo sapiens Adult MZ; Gruta del Indio GIRA-831 7364 Apatite -10.5 Homo sapiens MZ; Gruta del Indio JP/J4 7346 Enamel -13.5 Homo sapiens 2100-1700 MZ; Jaime Prats JP/J4 7347 Collagen -17.4 9.8 Homo sapiens 2100-1700 MZ; Jaime Prats JP/J4 7348 Apatite -13.5 Homo sapiens 2100-1700 MZ; Jaime Prats JP-1155 6219 Collagen -16.8 10.6 Homo sapiens 2100-1700 F 20-26 yrs MZ; Jaime Prats JP-1155 6220 Apatite -10.2 Homo sapiens 2100-1700 F 20-26 yrs MZ; Jaime Prats JP-1155 6221 Enamel -8.6 Homo sapiens 2100-1700 F 20-26 yrs MZ; Jaime Prats JP-1352 7338 Enamel -11.2 Homo sapiens 2100-1700 MZ; Jaime Prats JP-1352 7339 Apatite -10.6 Homo sapiens 2100-1700 MZ; Jaime Prats JP-1352 7340 Collagen -16.3 9.9 Homo sapiens 2100-1700 F? MZ; Jaime Prats MGA-1 6214 Collagen -14.2 10.9 Homo sapiens MZ; RQ-1 MGA-1 6215 Apatite -8.9 Homo sapiens MZ; RQ-1 MGA-1 6216 Enamel -8.7 Ho mo sapiens MZ; RQ-1 R-A-1 5905 Apatite -10.7 Lama guanicoe R-A-1 6170 Collagen -19.0 4.3 Lama guanicoe R-A-1 7368 Faunal -14.2 9.0 Lama guanicoe SJ; Angualasto R-A-10 5913 Apatite -8.9 Lama guanicoe R-A-10 6179 Collagen -18.8 4.3 Lama guanicoe R-A-11 5914 Apatite -11.5 Pterocnemia pennata 118

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R-A-11 6180 Collagen -21.0 4.9 Pterocnemia pennata R-A-2 5906 Apatite -6.8 Lama guanicoe R-A-2 6171 Collagen -14.7 5.0 Lama guanicoe R-A-2 7369 Faunal -18.3 6.7 Rhea americana SJ; Angualasto R-A-3 5907 Apatite -11.1 Lama guanicoe R-A-3 6172 Collagen -19.4 4.6 Lama guanicoe R-A-3 7370 Collagen -18.1 5.6 Lama guanicoe SJ; Morrillos R-A-4 6173 Enamel -9.1 Lama guanicoe R-A-5 5908 Apatite -11.5 Cholephaga melanoptera R-A-5 6174 Collagen -22.0 4.1 Cholephaga melanoptera R-A-6 5909 Apatite -11.8 Rhea americana R-A-6 6175 Collagen -20.0 5.7 Rhea americana R-A-7 5910 Apatite -12.1 Pterocnemia pennata R-A-7 6176 Collagen -20.6 4.6 Pterocnemia pennata R-A-8 5911 Apatite -9.1 Lagidium viscacia R-A-8 6177 Collagen -19.3 3. 7 Lagidium viscacia R-A-9 5912 Apatite -11.1 Chaetophractus villosus R-A-9 6178 Collagen -17.7 5.6 Chaetophractus villosus R-V-1 6181 Botanical -9.7 3.4 Zea mays R-V-10 6190 Botanical -20.8 Geoffroea decorticans R-V-11 6191 Botanical -24.9 11.6 Prosopis sp. R-V-13 6193 Botanical -24.4 1.6 Schinus polygamus R-V-2 6182 Botanical -9.6 3.9 Zea mays R-V-3 6183 Botanical -23.2 13.1 Cucurbita maxima R-V-4 6184 Botanical -25.4 Lagenaria sp. R-V-5 6185 Botanical -27.6 6.9 Chenopodium sp. 2200 MZ; GrN 5398 R-V-6 6186 Botanical -23.9 Prosopis sp. R-V-6 7376 Botanical -24.3 7.0 Cucurbita maxima SJ; Calingasta 119

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R-V-7 6187 Botanical -25.4 1.6 Cassia arnottiana R-V-8 6188 Botanical -24.0 5.5 Phaseolus vulgaris R-V-9 6189 Botanical -20.2 14.0 Geoffroea decorticans R-V-9 7379 Botanical -24.2 9.8 Cucurbita maxima SJ; Iglesia SJ10-ENT1 7157.1 Hair -13.4 9.6 Homo sapiens 600 F SJ; Angualasto SJ10-ENT1 7157.2 Hair -12.7 9.0 Homo sapiens 600 F SJ; Angualasto SJ10-ENT1 7157.3 Hair -13.6 8.6 Homo sapiens 600 F SJ; Angualasto SJ10-ENT1 7157.4 Hair -13.2 8.6 Homo sapiens 600 F SJ; Angualasto SJ10-ENT1 7157.5 Hair -13.9 8.0 Homo sapiens 600 F SJ; Angualasto SJ10-ENT1 7157.6 Hair -15.1 8.7 Homo sapiens 600 F SJ; Angualasto SJ10-ENT1 7157.7 Hair -16.6 9.4 Homo sapiens 600 F SJ; Angualasto SJ10-ENT1 7157.8 Hair -15.6 9.6 Homo sapiens 600 F SJ; Angualasto SJ10-ENT1 7158 Skin -14.1 12.0 Homo sapiens 600 F SJ; Angualasto SJ10-ENT1 7159 Apatite -8.2 Homo sapiens 600 F SJ; Angualasto SJ10-ENT1 7386 Collagen -12.3 9.9 Homo sapiens 600 F SJ; Angualasto SJ1-ENT7 7103 Skin -16.4 11.8 Homo sapiens 7900-4200 F SJ; Gruta Morrillos SJ1-ENT7 7104 Apatite -13.1 Homo sapiens 7900-4200 F SJ; Gruta Morrillos SJ1-ENT7 7381 Collagen -17.3 9.7 Homo sapiens 7900-4200 F SJ; Morrillos Gruta 1(F) SJ2 7105.1 Hair -14.9 10.6 Homo sapiens 800 SJ; Calingasta SJ2 7105.2 Hair -13.4 10.2 Homo sapiens 800 SJ; Calingasta SJ2 7105.3 Hair -13.5 10.4 Homo sapiens 800 SJ; Calingasta SJ2 7105.4 Hair -14.4 9.6 Homo sapiens 800 SJ; Calingasta SJ2 7106 Apatite -10.1 Homo sapiens 800 SJ; Calingasta SJ2 7107 Skin -14.4 13.5 Homo sapiens 800 SJ; Calingasta SJ2 7108 Enamel -6.3 Homo sapiens 800 SJ; Calingasta SJ2 7382 Collagen -13.8 9.5 Homo sapiens 800 SJ; Calingasta SJ3-ENT3 7109.1 Hair -15.3 9.2 Homo sapiens 590 SJ; Punta del Barro SJ3-ENT3 7109.2 Hair -16.2 9.7 Homo sapiens 590 SJ; Punta del Barro 120

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SJ3-ENT3 7109.3 Hair -16.9 9.7 Homo sapiens 590 SJ; Punta del Barro SJ3-ENT3 7109.4 Hair -16.6 9.4 Homo sapiens 590 SJ; Punta del Barro SJ3-ENT3 7109.5 Hair -15.6 9.6 Homo sapiens 590 SJ; Punta del Barro SJ3-ENT3 7110 Apatite -9.8 Homo sapiens 590 SJ; Punta del Barro SJ3-ENT3 7111 Collagen -13.3 9.5 Homo sapiens 590 SJ; Punta del Barro SJ3-ENT3 7112 Enamel -9.0 Homo sapiens 590 SJ; Punta del Barro SJ3-ENT3 7383 Skin -16.3 11.6 Homo sapiens 590 SJ; Punta del Barro SJ4-ENT2 7141 Skin -15.3 13.1 Homo sapiens 640 SJ; Angualasto SJ4-ENT2 7142 Apatite -10.3 Homo sapiens 640 SJ; Angualasto SJ4-ENT2 7384 Collagen -13.8 10.1 Homo sapiens 640 SJ; Angualasto SJ5-ENT2 7143 Enamel -8.3 Homo sapiens 880 SJ; C Calvario SJ5-ENT2 7144 Skin -18.0 14.0 Homo sapiens 880 SJ; C Calvario SJ5-ENT2 7145.1 Hair -15.0 10.3 Homo sapiens 880 SJ; C Calvario SJ5-ENT2 7145.2 Hair -15.2 10.3 Homo sapiens 880 SJ; C Calvario SJ5-ENT2 7145.3 Hair -15.5 8.7 Homo sapiens 880 SJ; C Calvario SJ5-ENT2 7145.4 Hair -15.2 9.1 Homo sapiens 880 SJ; C Calvario SJ5-ENT2 7145.5 Hair -17.1 9.3 Homo sapiens 880 SJ; C Calvario SJ6-ENT8 7146.1 Hair -19.4 9.0 Homo sapiens 2000 F Adult SJ; Gruta 1 MorrillosAnsilta SJ6-ENT8 7146.2 Hair -20.1 9.5 Homo sapiens 2000 F Adult SJ; Gruta 1 MorrillosAnsilta SJ6-ENT8 7146.3 Hair -20.6 9.6 Homo sapiens 2000 F Adult SJ; Gruta 1 MorrillosAnsilta SJ6-ENT8 7146.4 Hair -20.8 9.3 Homo sapiens 2000 F Adult SJ; Gruta 1 MorrillosAnsilta SJ6-ENT8 7146.5 Hair -20.2 8.8 Homo sapiens 2000 F Adult SJ; Gruta 1 MorrillosAnsilta SJ6-ENT8 7147 Skin -19.8 17.4 Homo sapiens 2000 F Adult SJ; Gruta 1 Morrillos SJ6-ENT8 7380 Apatite -14.0 Homo sapiens 2000 SJ; Gruta 1 MorrillosAnsilta SJ6-ENT8 7385 Collagen -17.3 8.1 Homo sapiens 2000 SJ; Gruta 1 MorrillosAnsilta SJ7-ENT2 7148.1 Hair -16.8 10.2 Homo sapiens 4070 M Adult SJ; Gruta 1 Morrillos-Morrillos SJ7-ENT2 7148.2 Hair -16.7 9.7 Homo sapiens 4070 M Adult SJ; Gruta 1 Morrillos-Morrillos SJ7-ENT2 7148.3 Hair -17.3 9.4 Homo sapiens 4070 M Adult SJ; Gruta 1 Morrillos-Morrillos 121

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SJ7-ENT2 7149 Skin -20.9 16.7 Homo sapiens 4070 M Adult SJ; Gruta 1 Morrillos-Morrillos SJ7-ENT2 7150 Enamel -9.2 Homo sapiens 4070 M Adult SJ; Gruta 1 Morrillos-Morrillos SJ7-ENT2 7151 Apatite -12.2 Homo sapien s 4070 M Adult SJ; Gruta 1 Morrillos-Morrillos SJ7-ENT2 7152 Collagen -15.3 10.8 Homo sapi ens 4070 M Adult SJ; Gruta 1 Morrillos-Morrillos SJ8-ENT5 7153.1 Hair -14.2 9.0 Homo sapiens 2000 M SJ; Gruta 1 MorrillosAnsilta SJ8-ENT5 7153.2 Hair Homo sapiens 2000 M SJ; Gruta 1 MorrillosAnsilta SJ8-ENT5 7153.3 Hair -19.4 7.4 Homo sapiens 2000 M SJ; Gruta 1 MorrillosAnsilta SJ8-ENT5 7153.4 Hair -20.0 7.8 Homo sapiens 2000 M SJ; Gruta 1 MorrillosAnsilta SJ8-ENT5 7153.5 Hair -19.8 8.6 Homo sapiens 2000 M SJ; Gruta 1 MorrillosAnsilta SJ8-ENT5 7154 Skin -17.6 12.2 Homo sapiens 2000 M SJ; Gruta 1 MorrillosAnsilta SJ9-ENT1 7155 Skin -11.6 12.3 Homo sapiens 1400-1200 F Adult SJ; Hilario SJ9-ENT1 7156.1 Hair -10.7 10.0 Homo sapiens 1400-1200 F Adult SJ; Hilario SJ9-ENT1 7156.2 Hair -12.9 9.0 Homo sapiens 1400-1200 F Adult SJ; Hilario SJ9-ENT1 7156.3 Hair -13.4 9.2 Homo sapiens 1400-1200 F Adult SJ; Hilario SJ9-ENT1 7156.4 Hair -16.5 9.6 Homo sapiens 1400-1200 F Adult SJ; Hilario SJ9-ENT1 7156.5 Hair -10.6 9.6 Homo sapiens 1400-1200 F Adult SJ; Hilario SJ9-ENT1 7156.6 Hair -11.0 9.5 Homo sapiens 1400-1200 F Adult SJ; Hilario SJ9-ENT1 7156.7 Hair -10.3 9.1 Homo sapiens 1400-1200 F Adult SJ; Hilario SJ9-ENT1 7156.8 Hair -10.0 9.5 Homo sapiens 1400-1200 F Adult SJ; Hilario 122

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123 Appendix C. Bone Apatite Sample Processing Form Name of Sample Series _____________________ Lab Worker_____________________ Fill in the date for each item below. Use a sep arate sheet for each batch of samples processed. _____ 1. Select approximately 1 gram of whole bone. _____ 2. Clean bone physically and then u ltrasonically to remove dirt and adherent materials. Use brushes and dental tools as necessary. _____ 3. Dry bone in drying oven. _____ 4. Pulverize bone using ball mill or use drill to produce bone powder. _____ 5. Pass bone powder through mesh scr een stack to separate size fractions (if necessary). _____ 6. Weigh out approximately 10 mg of bone powder into a 1.5 ml centrifuge tube. Label with USF number. _____ 7. Add 1 ml of 2% bleach solution to remove collagen, bacterial proteins, humates. Let stand for 72 hours. _____ 8. Centrifuge the samp le and pour off bleach solution using pipette if necessary to remove solution without losing sample. Replace with distilled water. Repeat 4 times. _____ 9. Dry bone powder in drying oven. Weigh sample. _____ 10. Pre-treat bone with 1 ml of 1 M acetic acid/sodium acetate buffer solution for 24 hours. _____ 11. Centrifuge the sample, pour off ace tic acid/sodium acetate solution using pipette if necessary to remove solution without losing sample. Replace with distilled water. Repeat 4 times. _____ 12. Dry bone powder in drying oven. Weigh sample. USF # Lab/Museum # Initial Weight Weight after bleach Weight after acetic acid Run Weight

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124 Appendix D. Tooth Enamel Sample Processing Form Name of Sample Series _____________________ Lab Worker_____________________ Fill in the date for each item below. Use a sep arate sheet for each batch of samples processed. _____ 1. Select tooth sample. _____ 2. Clean tooth physically and then u ltrasonically to remove dirt and adherent materials. Use brushes and dental tools as necessary. _____ 3. Dry tooth in drying oven. _____ 4. Drill enamel powder from tooth surface being careful not to reach dentin _____ 5. Weigh out approximately 10 mg of enamel powder into a 1.5 ml centrifuge tube. Label with USF number. _____ 6. Add 1 ml of 2% bleach solution to remove collagen, bacterial proteins, humates. Let stand for 24 hours. _____ 7. Centrifuge the samp le and pour off bleach solution using pipette if necessary to remove solution without losing sample. Replace with distilled water. Repeat 4 times. _____ 8. Dry enamel powder in drying oven. Weigh sample. _____ 9. Pre-treat enamel with 1 ml of 1 M acetic acid/sodium acetate buffer solution for 24 hours. _____ 10. Centrifuge the sample, pour off ace tic acid/sodium acetate solution using pipette if necessary to remove solution without losing sample. Replace with distilled water. Repeat 4 times. _____ 11. Dry bone powder in drying oven. Weigh sample. USF # Lab/Museum # Initial Weight Weight after bleach Weight after acetic acid Run Weight

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125 Appendix E. Bone Collagen Sample Processing Form Name of Sample Series _____________________ Lab Worker_____________________ Fill in the date for each item below. Use a sep arate sheet for each batch of samples processed. _____ 1. Select approximately 1 gram of w hole bone and put in labeled jar with lid. _____ 2. Clean bone physically and then u ltrasonically to remove dirt and adherent materials. Use brushes and dental tools as necessary. _____ 3. If bone was treated with preservative, rinse or soak in acetone. _____ 4. Dry bone in drying oven. Weigh sample. _____ 5. Soak in 50 mls 0.1 M NaOH (4g per liter) for 24 hours to remove humic acids. _____ 6. Pour off NaOH and rinse thoroughly with distilled water. Add 50 mls 2% HCl (200 mls concentrated HCL, 3.6 liters di stilled water) to re move bone mineral. _____ 7. Cut or pulverize bone into smaller pieces as necessary. _____ 8. Replace HCl solution with fresh acid after 24 hours. _____ 9. Replace HCl again after another 24 hours. Repeat as necessary until bone is totally demineralized. _____ 10. Pour off HCl acid solution and rin se thoroughly with distilled water. Add 50 mls 0.1 M NaOH (4 g per liter) and soak 24 hours to remove humic acids. _____ 11. Pour off NaOH solution and rinse thoroughly with distilled water. Add 50 mls defatting solution (2:1:0.8 mixture of me thanol, chloroform and distilled water), and soak 24 hours to remove fat content. _____ 12. Pour off defatting solution into waste jars; rinse extra thoroughly. Transfer samples to 2-dram vials, and label with indelible marker. _____ 13. Oven-dry samples. _____ 14. Weigh samples and calculate collagen yield. USF # Lab/Museum# Initial Weight Final Weight % Yield Run Weight

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126 Appendix F. Hair Sample Processing Form Name of sample series ____________________ Lab worker responsible____________________ Fill in the date for each item below. Use a sep arate sheet for each batch of samples processed. ____ 1. Put on latex gloves and a hair net or ball cap ____ 2. Clean several large test tubes using distilled water ____ 3. Clean a fairly large beaker by rinsi ng it with acetone and then with distilled water ____ 4. Select a hair sample ____ 5. Line up approximately 15 strand s of hair (record number of strands) ____ 6. Cut into 2 cm sections along the shaft; distilled water may be used if necessary to dampen the hair for easier handling ____ 7. Label the test tube with sample identifier (.1,.2,.3, etc. of their lab number) ____ 8. Place the hair sample in the test tube ____ 9. Fill the test tube with distilled water ____ 10. Place the test tube upright in the large beaker, place it in the sonication unit ____ 11. Repeat steps 4 through 10 until all the samples are prepared ____ 12. Fill the sonication unit to a level belo w the top of the test tubes with distilled water ____ 13. Sonicate the samples for 15 minutes at room temperature ____ 14. Remove the samples from the sonication unit ____ 15. Remove a test tube and carefully pour out the liquid while keeping the hair inside the tube ____ 16. Put on a mask and fill the test tube with a 2:1 v/v mixture of methanol and chloroform ____ 17. Place the filled tube upright in a large beaker. When the beaker is full, place it in a sonication unit ____ 18. Repeat steps 13 through 17 for the rest of the samples ____ 19. Ensure the sonication unit is filled to a level below the top of the test tubes with distilled water ____ 20. Sonicate the samples for 15 minutes at room temperature ____ 21. Replicate the methanol/chloroform cleansing by repeating steps 15 through 20 (two total methanol:chloroform rinses) ____ 22. Remove a test tube and carefully po ur out the methanol/chloroform mixture while keeping the hair inside the tube ____ 23. Fill the tube with distilled water. ____ 24. Place the tube upright in a large beak er. When the beaker is full, place it in the sonication unit ____ 25. Repeat steps 22 through 24 until all samples are prepared ____ 26. Fill the sonication unit to a level belo w the top of the test tubes with distilled water ____ 27. Sonicate the samples for 15 minutes at room temperature ____ 28. Remove the samples from the sonication unit ____ 29. Remove each test tube and carefully po ur out the distilled water while keeping the hair inside the tube. Place the tubes upright in a large beaker ____ 30. Put the samples in a safe area with circulating air, such as a chemical hood ____ 31. Allow the samples to air dry for at least 48 hours ____ 32. Examine microscopi cally to ensure cleanliness


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Before the Inca :
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2006.
3 520
ABSTRACT: A dietary reconstruction was performed in order to understand changing prehistoric subsistence patterns in the Central Andean geographical area of the Argentine Cuyo that includes the provinces of San Juan and Mendoza. Archaeologically, the Cuyo is also known as a boundary between Andean agriculturalists and the foragers of Patagonia. One hypothesis being tested is whether this area was one of the last South American cultural groups to convert to maize cultivation, probably around 2000 BP. The process of stable isotope analysis is used to reconstruct the diets of individuals, as it reveals the relative proportions of C3 and C4 plants and the contribution of aquatic resources to otherwise terrestrial diets, as well as variations in trophic level of the foods consumed.In this study the bones, teeth, hair, and flesh from 45 individuals were tested to address specifically total and protein diets, as well as seasonal variation and changes between childhood and adulthood. This process, when used in combination with previous analyses, such as midden or faunal analysis, allows researchers to evaluate the results of those previous studies, and thus compose a more thorough reconstruction of the lifestyles of a prehistoric culture.Information garnered from this study indicates that the times of dietary transition were variable, with seasonal patterns becoming more stable over long periods. Furthermore, some members of the study population demonstrate the existence of nutritional stress indicators, such as dental caries, that can be viewed in relation to the dietary shifts that may have been a cultural adaptation to the environment of the Cuyo. Overall, this study shows the early adoption of maize agriculture in central western Argentina and recommends future studies that analyze the relationships between agriculture, diet, and nutrition in the New World.
502
Thesis (M.A.)--University of South Florida, 2006.
504
Includes bibliographical references.
516
Text (Electronic thesis) in PDF format.
538
System requirements: World Wide Web browser and PDF reader.
Mode of access: World Wide Web.
500
Title from PDF of title page.
Document formatted into pages; contains 126 pages.
590
Adviser: Robert H. Tykot, Ph.D.
653
Archaeology.
Archaeometry.
Bone chemistry.
Diet.
Environment.
Human ecology.
Mendoza.
Mummification.
Nutrition.
San Juan.
South America.
Stable isotope analysis.
690
Dissertations, Academic
z USF
x Applied Anthropology
Masters.
773
t USF Electronic Theses and Dissertations.
4 856
u http://digital.lib.usf.edu/?e14.1588