Giardia lamblia in White nosed Coatis Sha 1 White nosed Coatis ( Nasua narica) as Hosts for the Endoparasite Giardia lamblia Megan Sha Department of Ecology and Evolutionary Biology University of California, Los Angeles EAP Tropical Biology and Conservation Program, Fall 2016 16 December 2016 ABSTRACT Giardiasis the disease caused by the protozoan endoparasite, Giardia lamblia has become an increasingly concerning threat to both animals and humans, especially for travelers in Parasitic infections are an omnipresent problem that are ofte n a result of zoonotic diseases, or diseases that are transmitted from animal to human. This study strove to determine if white nosed coatis ( Nasua narica) could potentially be hosts to Giardia lamblia in Monteverde similar to how they are capable of acti ng as hosts for other zoono ses. I collected 28 coati fecal samples at three locations within Monteverde, Costa R ica, and found five samples positive for Giardia lamblia Not only did this demonstrate that Nasua narica could act as hosts for the endoparasite, but the individual s surveyed were also hosts to 19 othe r parasitic species ranging from roundworms to tapeworms S uch parasitic incide nces depended on the locations in which t he samples were found, along with host s interaction s with possible sources of contamination in the particular environment; the latter could have resulted from contact with infes ted water and/or food, or through fecal oral routes. Close human proximity to coatis can pose a danger to human healt h and increase our risks for contracting the disease while jeopardizing the health of coatis and neighboring wildlife as well Pizotes ( Nasua narica ) como Hospederos del E ndoparsito Giardia lamblia RESUMEN Giardiasis la enfermedad causada por el protozoario parsito Giardia lamblia se ha convertido en una amenaza creciente tanto para animales como para humanos. Las infecciones por parsitos son un problema omnipresente que a menudo resultan en zoonosis, enfermedades transmitidas de animales a humanos. Este estudio evalu si los pizotes ( Nasua narica ) son huspedes potenciales de Giardia lamblia en Monteverde, as como de otras zoonosis. Recolect 28 muestras fecales de pizotes en tres sitios en Monteverde, Costa Rica y encontr cinco muestras positivas para Giardia lamblia No solo esto demuestra que Nasua narica acta como husped del endoparsito, sino que tambin de otras 19 especies que van desde gusanos redondos a planos, resultados de sus interacciones con otro s huspedes, fuentes de agua o de comida expuestas a contaminacin fecal. La cercana a los humanos y por ser animales silvestres hacen de los pizotes un factor de riesgo a la salud humana y la de otros animales tambin. Parasitism is a symbiotic relati onship between two organisms in which a parasite lives off a ho st and derives benefits from it, while harmin g and possibly killing the host in the process.
Giardia lamblia in White nosed Coatis Sha 2 Two main para sitic relationships exist : ectoparasitism and endoparasitism, in which the parasites re side either externally or internally of t heir hosts, respectively. In particular, e ndop arasites can perpetuate between hosts via zoonotic or vector borne transmission, or through the ingestion of contaminated water and/ or food sources (Bar yam, 2011). Zoonotic diseases are particularly concerning, since infected animals are capable of transferring their diseases to humans. From 2006 to 2010, Costa Rica reported ov e r 84,000 cases of dengue fever and high morbidity rates from leptospirosis which numbered up to 46 deaths; both of these diseases are zoonoses (Pan American Health Organization, 2012). Emerging zoonoses, such as Giardiasis, are especially crucial to control in developing countries, where health services may not be as advanced as those in devel oped countries White nosed coatis, or Nasua narica, were chosen as the target species for this study because of their abundance in Monteverde, Costa Rica, and the close interactions they h ave with the human residents and domestic animals living there The ir constant presence amongst humans may be a concerning issue if their potential for carrying a zoonosis such as Giardia lamblia is confirmed. Giardiasis is a disease caused by a protozoan endoparasite, Giardia lamblia, and affects 3 5% of individuals who travel to Latin or Central America (I A MAT, 2016) Its detriment to travelers has grown more profound with time, since it and other protozoal agents account for Costa Rica (ibid) The parasite thrives in warm, humid climates, and is most easily contracted by both humans and animals through the ingestion of contaminated water from lakes and streams, or from infected food; fecal oral routes are a common transmission pathway as well. The protozoa cause acute or chronic diarrhea with loose and foul smelling mucous stools, and require a host ranging anywhere from pe ople to wild animals to survive (CFSPH, 2012). Within seven to fourteen days of parasite manifestatio n, the host can begin displaying symptoms and shed inactive cysts through its feces. While Giardia lamblia does not manifest within a host with the intention to kill, hosts can often fall victim to death from shock or dehydration if the fluids and electrolytes lost from diarrhea are not replaced soon enough (CSFPH, 2012). Because cysts can remain viable for several weeks or even months in cold conditions, disease transmission through the ingestion of cysts is much more common as opposed to ingestion mobile form, the trophozoites; the latter can only last briefly in feces once they are exposed to the environment. Upon ingestion, the cysts move to the intestines and are lysed open, or split, via excystation, producin g two trophozoites that then multiply through longitudinal binary certain number of divisions have elapsed and the trophozoites have passed through the small intestine, bile salts and other substances along the colon stimulate trophozoites encystation, where the protozoa begin to form cell walls. The newly formed cysts are passed through the anus and into the surrounding environment for further tran smission (CDC, 2015). White nosed coat is, or Nasua narica are potential animal reservoirs for Giardi asis and can be infected from an ingestion of just one cyst (Gompper et al., 1996). These diurnal mammals live in large social groups, or bands, and display reciprocal altruism through grooming While they are primarily found in upper elevations and mountain forests, they are also present in urban ar eas such as Monteverde where they adapt quickly to human presence and forage closely to food abundant areas, such as compost piles. White nosed coa tis are omnivorous ground foragers, feeding on small vertebrates, fruits, insects, and even carrion. Females and young coatis travel in bands which makes the likelihood for their exposure to cyst infected feces very high if
Giardia lamblia in White nosed Coatis Sha 3 there is even one incidence of disease. M ales are solitary and leave their band once they have matured, and are only accepted into others during mating seasons before they are excluded once again. In Monteverde, coatis are primed as favo rable reservoirs for Giardiasis, as they are constantly surrounded by mammalian, coati, and canine feces, all of which may potentially hold Giardia cysts A study focusing on canines at San I sidro de El General, Costa Rica surveyed fecal and serum samples concluding that the prevalence of zoonotic parasites was high in domestic animals, with 75% of the dogs surveyed being hosts to Giardia lamblia among other diseases (Scorza et al., 2011). Personal observations indicate that stray dogs share the same ranges as coatis, suggesting that coatis can co ntract Giardiasis from infected canines, as well as through with va rious different environmental vectors diseases, such as Trypanosoma cruzi (Mehrkens et al., 2013), led me to believe that their capacity for carrying Giardia lamblia is plausible. Over the course of my two wee k s tudy, my main question was: Can white nosed coatis be hosts for the endoparasite, Giardia lambl ia? However, I also investigated the fol lowing : What other parasites might white nosed coatis be hosts for ? MATERIALS AND METHODS From 22 November to 1 December, 2016, I surveyed three sites in Monteve rde, Costa Rica: Estaci n Biol gica, Crandell Memorial at Monteverde Institute, and Bajo del Tigre. During the first five days I alternated between the days that I surveyed a t the Station and the Institute Since I was only able to find one sample at Crandell Memorial that entire week, I halted surv eying around the Institute and changed to surveying solely around Estacion Biologica and Bajo del Tigre for the sake of collecting as many samples I could within the time constraints of this study I surveyed for about two to three hours each morning at th e respective locations and either tried following coatis until they defecated and ins pecting trails that I knew coatis frequented to search for feces that had been dep osited earlier. If I were unable to observe coatis defecate due to weather, time, or other limitations, I was 85 % confident that the feces belonged to my target species since diameter s ranged from 0.8 2.5cm and displayed evid ence of seeds evidence supporting their tendency to feed on fruit (Figure 1 ) (El broch, 2003) ; I also assumed that each fecal sample I collected belonged to a different coati individual. I afterwards into an excel sheet. Each sample was also grade d using the Bristol Stool Chart, which gave each sample a label ranging from Type 1, (Riegler and Esposito, 2001).
Giardia lamblia in White nosed Coatis Sha 4 Figure 1. Example of a fecal sample from the Station with a diameter of 2.0cm and seeds present. After each collection period, I to ok my samples to the lab to perform two direct wet mount fecal staining s I used two types of fecal direct smears that targeted for motile and no n motile parasitic stages in semi formed or loose to fluid feces (Nolan, 2016). I prepped a Saline stained slide for each sample; the former stained for trophozoites while the latter stained for cysts (ibid) To observe for the motile f orm, I prepared a 0.85% NaCl solution by dissolving 0.85g of NaCl in 100mL of water and used a plastic pipette to drop one to three drops of solution onto a smear of feces that I had prepped on a slide beforehand (ibid) I used a sterile wooden applicator to mix the feces and solution together until the mixture was uniform, and placed a coverslip over it afterwards. To prep for cyst staining, I conducted the exact same iginal 20% solution, instead ( University of Tennessee 2009). I looked at all my samples under the microscope at 4 0 x, 1 0 0x, and 40 0 x magnifications, and noted down all cyst and/or parasitic structures in my slides by making drawings or taking pictures of the images through the microscope eyepiece. I then used these images to make a key of all the parasitic species and morphospecies th at might be present. I identified for Giardia lamblia cysts and trophozoites based on the following descriptions of the two: the cysts are egg shaped and measure 8 14m in length by 7 10m in width, and have the following structures after developing into m ature cysts: four nuclei, four median bodies, two axostyles or axonemes that bisect the cyst in length, and eight pairs of flagella that surround or bind to the axostyles (Liu and Nevins, 2009). If the cysts are not fully matured, they only display half th e number of all the above structures. The trophozoites appear as bilaterally symmetrical, pear shaped protozoans with broad anterior and narrow posterior sides that measure 10 12m in length by 5 7m in width. They have two nuclei, a large disk at the ant erior, two axonemes, and four pairs of flagella that extend out to assist in movement. I also identified other parasites that might be present in Giardia by taking pictures of them under the microscope at a 400x magnification and comparing their structures to known parasitic parasite morphologies (Table 1). Lastly, I performed two Chi Squared Tests of Independence on Microsof t Excel to deter mine if: 1) the significance of Giardia lamblia incidences depended on the locations the fecal samples containing the parasite were found at, and 2) if there might be differences in the frequencies of Giardia between the sites the cysts were found at To clean my workspace, I wiped the microscope slid es and coverslip cleanly of fecal matter using paper towels, disinfected them with 70% ethanol, rinsed them under the sink with
Giardia lamblia in White nosed Coatis Sha 5 wate r, and left them out to air dry The saline and iodine solutions were discarded in the sink, and the leftover fecal samples in the plastic bags were either returned to the forest or flushed down the toilet while the plastic bags were discarded in the trash bin in the lab. RESULTS I found p arasites in all 28 fecal samples and five positive samples for Giardia lamblia cysts Scats were only found on a fraction of the days that I surveyed for samples, so the numbers of samples found at each location were unequal. I collected 25 samples from Estacion Biologica, one from Crandell Memorial, and two from Bajo del Tigre. Three of th e samples containing Giardia lamblia were found at the Station, two were found at Bajo del Tigre. While the abundance s of the other parasites were not accounted for, a total of 20 different parasitic morphospecies was also recorded, with Entamoeba spp. bei ng the most prevalent and appearing in 21 of the 28 total individuals (Table 1). Trichuris spp. and Blastocystis hominis were both the second most prevalent parasites and were found in ten samples. Others, such as Diphyllobothrium latum and Toxocara cati, were each only found in one sample. Although I found no Giardia lambli a trophozoites in any of my samples, I identified four of the five samples as fully m atured cysts and the last sample as a juvenile cyst based on the criteria listed in my methods and co mparing my images to known images of Giardia (Kolarova et al., 2000). I took all the pictures at a magnification of 400x (Table 1), but only Samples 2, 6 and 20 were clear enough to take pictures of. Samples 2 6 10 and 11 were identified through evidence of two axonemes and four median bodi es within the cysts (CDC, 2013). Meanwhile, Sample 20 was identified by observing a single axoneme at the posterior end of the cyst and also what appeared to be nuclei at the anterior part of the cyst. T hree s amples with Giardia lamblia (Samples 2, 10 an d 11) were found at the Station and two (Samples 6 and 20) were found at Bajo del Tigre. The first Chi Squared Test I conducted revealed that Giardia lamblia incidences depended on where the respective fecal samples were found at (X 2 =10.00, df =2, p value=0.0067). A second Chi Squared Test showed that there were no differences in the frequencies of Giardia incidence between the sites the parasites were found at (X 2 =0.23, df =2, p value= 0.89). Asides from Gia rdia, I also found 19 other parasites (Figures 2 and 3). Morphospecies B appears to be Blastocystis hominis one of the most common intestinal protozoan parasites around the world (Figure 2) (Lehman, 2011). Morphospecies C resembles a Diphyllobothrium latum a tapeworm, while D appears to be a Toxocara cati roundworm cyst (A r cari et al., 2000). Morph E was circular with multiple nuclei that resembled the roundworm, Entamoeba spp, and was observed in the greatest number of samples (Figure 2) (ibid) Morp ho F resembles Ascaris lumbricoides, and supports previous literature stating that Nasua narica cou ld be host to these parasites (Gompper et al., 1996) Morpho G was unclear, but could possibly be yea st and H was an unidentifiable roundworm egg (A r cari et al., 2000). Morpho I and L could both be Coccidia parasites, and L could either be Isospora spp., a Coccidia, or Hymenolepis spp. tapeworm. Based on picture comparisons, Morpho J could either be Ancylostoma caninum or a part of the duodenale species an d Morphs K, M through O, R, and T were unidentifiable based on the sources of literature that were available. Morph P resembled the roundworm, Strongyloides spp. (Lehman, 2011); in my s lides I saw Morpho P in its matured and juvenile forms, the latter of which displayed shorter and thicker bodies. Morphos Q could possibly be Trichuris vulpis or trichiura a whipworm with tapered ended cysts. This and Morpho B were the second most
Giardia lamblia in White nosed Coatis Sha 6 abundant parasites. Lastly, Morpho S resembled Enterobius vermicularis or pi nworms that appear very closely in morphology to Morpho P but have tapered posterior ends that measure a sixth of its entire body length. Table 1 depicts all the discovered parasites along with their respective pictures. Figure 2. P revalence of different, non worm parasitic morphospecies amongst all fecal samples. See Table 1 for the compiled list of morphospecies. Figure 3. P revalence of different worm parasite morphospecies amongst all fecal samples. See Table 1 for a compiled list of morpho species.
Giardia lamblia in White nosed Coatis Sha 7 Fecal sample numbers 2, 5 and 20 contained the greatest number of parasites, with each containing six diff erent parasite species (Figures 4 and 5). I found f ecal Samples 2 and 5 from the Statio n and Sample 20 from Bajo del Tigre. F ecal Samples 7 and 8 reported zero parasites since the particles in the samples were too difficult to distinguish apart from fecal matter. Figure 4: Number of different parasitic species found in each fecal sample. Displays abundances in Fecal Samples 1 to 14 Figure 5: A continuation from Figure 4. Number of parasitic species found in each sample. Displays abundances in Fecal Samples 15 to 28. 0 1 2 3 4 5 6 7 15 16 17 18 19 20 21 22 23 24 25 26 27 28 Number of Different Parasite Species in Each Fecal Sample Fecal Samples
Giardia lamblia in White nosed Coatis Sha 8 DISCUSSION Giardia sis is a commonly transmitted through environmental pathways and animal vectors as a zoonotic pathogen. This study has demonstrated that Nasua narica are capable hosts for Giardia lamblia, supporting the idea that these mammals are a species that can impinge on animal and h uman health. While the protozoa were not the most abundant parasites, they were still found in approximately 18% of the total number of samples, 12% of the samples found at Estacion Biologica and 100% of the samples found at Bajo del Tigre I ncidences of Giardia do depend on the locations the sampl es are found at Firstly, the greatest frequencies of Giardia could have been found at the Station, simply because my chances of finding the parasite were much greater with a large sample size of 25. Greater numb ers of infected samples found at the Station may also be du e to the fact that coatis prefer living at higher elevations and tend to congregate where food is abundant (Gompper et al., 1996) People residing at the Station produce large amounts of organic fo od waste, and personal observations have revealed frequent visitations to s uc h compost piles by the coatis. M any are unafraid of humans and have even learn ed to associate people with food. This learned behavior could hold true for all coatis in Monteverde, b ut more common ly so a t the Station since compost piles are placed in the open, unlike at Crandell Memorial, for example, w here organic waste is place d in a shed away from wildlife Three solitary mal es and a troop of around twenty fi ve female and young coatis were spotted each day that I surveyed at the Station. Previous studies have shown that white nosed coatis prey on differing sources of food depending on what is most abundant during which seasons, and shi ft from consuming insects and arthropods or fruits when they are available, during the wet season s to fruits during the dry seasons (Alves Costa et al., 2004) Since Monteverde is currently transitioning to the dry season and many fruiting guava trees surround the Station, the fruits have naturally attracted many organisms, including coatis. Nasua narica tend to compensate for decreased insect prey abundances by concentrating foraging activity in a few select places with hi gh food availabilities (ibid). H ence, a significant portion of the samples originated fr om the Station, as opposed to Crandell Memorial or Bajo del Tigre, where food availability may not be as accessible This congregation phenomenon is not uncommon, seeing as Alves Costa et al. (2004) also reported cases where large groups of coatis number ing up to 70 individuals made an appearance upon discovering large compost piles Even though Giardia was found at the Station, only three of the 25 samples actually contained the parasite. One reason could be because the w ater that passes through this area via the Mquina Stream is cleaner than that used below this elevation. The Station is situated at an elevation of around 1541 meters, where exposure to contaminants might be much lower since this site is further away from urbanization. Meanwhile, fewer fecal samples we re found at Bajo del Tigre, possibly because this location did not contain as many acces sible food sources like at the S tation making it much more difficult to c ollect samples given the time constraints. Only two samples were found, but both exhibited Giardia Since Bajo del Tigre is at a lower elevation and is l ocated in a m ore urbanized location, the coatis there might be warier of people, cars, and dogs thus explaining my s mall sample size from this locality Giardia incidences for this particular area were still extremely high, however Bajo del Tigre is at a much lower elevation of 1387 meters and would be expected to contain a higher prevalence of Giardia because the bodies of water here would not only collect water flowing downwards from the Mquina Stream but also collect rain run off
Giardia lamblia in White nosed Coatis Sha 9 that could pick up Giardia cysts along the way down from the Station and deposit them into the streams below. Bajo del Tigre is locat ed much closer to such possible sources of water contamination, such as the Sucia Stream, Catarata Wate rfall, and Cuecha Stream, all of which pave way for higher risks of infection. Past studies in Canada noted that Giardia sis was most commonly contracted by drinking contaminated water, especially since Giardia cysts have be en found to survive for weeks or months in freshwater (Clayton, 2002). Crandell Memorial indicated no presence of Giardia but this might be because only one sample was foun d I did not see any coatis here, making it difficult to pre dict where the y might have defecate d at. If there had been more time to locate the feces in such a large area, more samples could have been analyzed to search for the presence of Giardia Since Crandell Me morial (1405 meters) is located directly next to the Cuecha Stream, I would have expected similar Giardia incidences to Bajo del Tigre if I had been able to obtain a larger sample size. Giardia lamblia is pr esent in Monteverde, but is ha rdly the most abundant parasite carried by Nasua narica since they can actually act as hosts for 19 other pa rasites. Entamoeba spp. cysts were transmission pathway resembles Giardia lamblia since it is generally transmitted through contami nated water or food sources, in addition to fecal oral routes. However, manifestation of this parasite usually results in symptomatic illnesses 80 98% of the time (Ericsson 2001) while Giardia has two main asse mblages, or infectious strains, A or B Hosts may or may not display symptoms depending on which assemblage is contracted, since assemblage A is associated with diarrhea while B is more asymptomatic and is often times overlooked when ind ividuals might actually have the disease (Haque, 2007) Two thirds of infected individuals with Giardia are unaware that they have the disease until about one to three weeks later, when acute symptoms mig ht finally begin These factors lead to fewer reported cases of Giardia than may actually exist. Thus, I may have just observed a lower prevalence of Giardia because the individuals were asymptomatic and had not yet begun excreting cysts in their feces during the given da ys I surveyed them at Trichuris vulpis and Blastocystis hominis were the second most abundant. The former develops in the intestines and primarily utilize s canines as hosts (Iowa State University, 2005) Many dogs roam free in Monteverde and even exchange direct encounters with coatis, meaning that dogs can pass Trichuris cysts easily in their feces and infect coatis via the feca l oral route. Blastocystis hominis is one of the most common intestinal parasites and thought to be transmitted through fecal ora route is still unknown (CDC, 2013) E xplanations for why select parasites mig ht be more or less abundant than others can be explained by preferences for specific temperatures or seasonal conditions. Fo r example, the prevalence of Entamoeba depend s completely on the regions and climates fecal samples are surveyed in in which c yst abundance s can range anywhere from 13.7% during dry seasons to 52% along the coasts (Bray et al., 1977) S harp rises in prevalence also correspond with bouts of rain, sugg e sting that cyst numbers are related with the wet season. Similarly, detection of Giardia cyst s nearly doubled during the rainy seasons compared to the dry seasons (Chuah et al., 2016) This could explain why Giardia was more abundant than 14 other parasites amongst my fecal samples, especially since rain from Hurricane Otto and from the days after could have easily provided more water routes of infection. Giardia lamblia or Entamoeba spp. have alternate ci rcuits of infection and are able to survive despite dry conditions while other parasites might display a significantly lower prevalence as a resul t of transmission constraints. S ome cysts
Giardia lamblia in White nosed Coatis Sha 10 may only be able to proliferate and divide in warm conditions, which the conditions in Monteverde were unable to meet during my study, resulting in a low er abundance for parasites such as Morphospecies G and O In addition, parasites may require access to specific hosts for certain stages of their life cycle a nd if these hosts were not present or easily accessible during transition season, this would have been as a limiting factor as well This study suggests that white nosed coatis are hosts to Giardia lamblia and a myriad of other endoparasites Because coatis live in close proximity to humans and domestic animals in Monteverde and utilize the same water sources precautions must be taken to reduce or eliminate the incidence of Giardia transmis sion from coati feces to human drinking water. People should avoid direc t contact with them and make sure their pets are checked for endoparasites and being bathed regularly Individuals should also strive to drink filter ed water, wash their hands before eating or drinking, and wash raw produce with filtered water If so much a s even o ne Giardia cyst is ingested, the person can be come infected. Recognition of the symptoms of Giardia in individuals is essential for not only the health of the infected individual, but for the community as a whole to maximize the avoidance of contaminated feces. F UTURE STUDIES If the given study period were longer, I could attempt to facilitate more acc urate results by attaining equal numbers of fecal samples from each location and ascertaining that each actually came from a unique individual by capturing my study subjects, marking them, collecting a fecal sample, and releasing them after This would be helpful for seeing if similar, high Giardia incidences might still be obtained at Bajo del Tigre, but with a greater sample size. If the results still hold true, they would contribute to the fact that disease incidences truly do depend on the locations at which fecal samples are found at. Further possibilities for analyzing incidences of Giardia lamblia could be to 1) survey for samples during the wet seasons and compare those results with those reported during the dry seasons to predict how transmission ra tes might change with response to varying levels of water sources, or 2) test at two different regions one near water and one located terrestrially to see if the idea that a higher protozoa prevalence near bodies of water is reinforced. ACKNOWLEDGEMENTS I would like to give a special thanks to my primary advisor, Federico Chinchilla for taking time out of his busy schedule to help me obtain my Bajo del Tigre permit so that I could survey in that area, helping obtain my materials needed for this study and for assisting me in the translation of my Abstract and providing boundless advice, patience, and reassurance in both the field and laboratory. A great thanks to my secondary advisor, Frank Joyce, as well, for encouraging me to pursue th is project idea from the very start, regardless of the doubts I may have had going forward. I would also like to thank Luisa and my friends in EAP for pointing out where all the coatis were at the Station whenever I was surveying the site and coul d not see m to sample I was able to find at Crandell Memorial for me. Lastly, a huge thanks to Raphael and others at Bajo del Tigre for permitting me to work there and, of course, Maureen Jimenez and the rest of my homestay family for welcoming me into their home as one of their own with such warmth and hospitality.
Giardia lamblia in White nosed Coatis Sha 11 LITERATURE CITED Alves Costa, C. P., Da Fon seca, G. A., & Christfaro, C. 2004 Variation in the d iet of the brown nosed coati ( Nasua nasua ) in southeastern Brazil. Journal of Mammalogy 85 (3), 478 482. Arcari, M. Boxendine, A., & Benette, E. 2009 Diagn osing medical parasites through coprological techniques Bar yam, Shlomiya. Evolution NECSI, 28 Feb. 2011. Bra y, R. S., & Harris, W. G. 1977 The epidemiology of infec tion with Entamoeba histolytica in the Gambia, West Africa. Transactions of the Royal Society of Tropical Medicine an Hygiene 71 (5), 401 407. C enters for Disea se Control and Prevention. 2013 Blast ocystis h ominis Retrieved from https://www.cdc.gov/dpdx/blastocystis/ Centers for Disease Control and Prevention. 2015 Paras ites Giardia Retrieved from https://www.cdc.gov/parasites/giardia/ Centers for Disea se Control and Prevention. 2013 Giardiasis: Laboratory Identification of Parasitic Diseases of Public Health Concern Retrieved from https://www.cdc.gov/dpdx/giardiasis/dx.html Chuah, C. J., Mukhaidin N., Choy, S. H., Smith, G. J., Mendenhall I. H., Lim, Y. A., & Ziegler, A. D. 20 16 Prevalence of Cryptosporidium and Giardi a in the water resources of the Kuang River catchment, Northern Thailand. Science of The Total Environment 562 701 713. Clayton, R. 2002 Giardia in Water Supplies: A Review of Current Knowledge. Foundation for Water Research. 1 24. Diagnostic Clinical Parasitology Service Laboratory. 2009 Antibody and Antigen Detec tion in Parasitic Infections: Detection of Parasitic Infections by Fecal Examination (Fifth ed.): 592 615. University of Tennessee College of Veterinary Medicine Elbroch, M. 2003 Mammal tracks & sign: a guide to North American species Stackpole books. Erics son, C. D., Steffen, R., & Okhuysen, P. C. 2001 Travel er's diarrhea due to intestinal protozoa. Clinical infectious diseases 33 (1), 110 114. Gompper, M. E., Gi ttleman, J. L., & Wayne, R. K. 1997 Gene tic relatedness, coalitions and social behaviour of white nosed coatis, Nasua narica Animal Behaviour 53 (4), 781 797. Haque, R. 2007 Human intestinal parasites. Journal of Health, Population and Nutrition 387 391. IAMAT ( International Association for Medical Assistance to Travelers ) 2016 Country Health Advice Costa Rica Costa Rica Overview. Iowa State University. 2005 Trichuriasis. The Center for Food Security and Public Health. 1 4. Kolarova, L V Nemeckova, D Zitova, and M Bednar. 2000 Images of Parasites. Charles University, Department of Medica l Microbiology. Parasitology
Giardia lamblia in White nosed Coatis Sha 12 Lehman, D 2011 Diagnostic Parasitology Images. University of Delaware, Department of M edical Laboratory Sciences. Liu, J and S Nevins. 2009 Giardia Morphology. Stanford University. Mehrkens, L. R., Shender, L. A., Yabsley, M. J., Shock, B. C., C hinchilla, F. A., Suarez, J., & Gilardi, K. V. 2013 White nosed coatis ( Nasua naric a ) are a potential reservoir of Trypanosoma cruzi and other potentially zoonotic pathogens in Monteverde, Costa Rica. Journal of wildlife diseases 49 (4), 1014 1018. Nolan, T 2006 Di rect Fecal Smear. Univ ersity of Pe nnsylvania School of Veterinary Medicine. Veterinary Parasitology Pan American Health Organization. 2012 Costa Rica. Health in the Americas Country Volume: 223 236. Riegler, G., & Esposito, I. 2001 Bristol scale stool form. A still valid help in medical practice and clinical research. Techniques in coloproctology 5 (3), 163 164. Scorza A. V., Duncan, C., Miles, L., & Lappin, M. R. 2011 Prevalence of selected zoonotic and vector borne agents in dogs and cats in Costa Rica. Veterinary parasitology 183 (1), 178 183. The Center for Food Security & Public Health (CFSPH) and Institute for International Cooperati on in Animal Biologies. 2012 Giardiasis 1 13.
Giardia lamblia in White nosed Coatis Sha 13 APPENDICES Table 1 : Identity of the d ifferent parasitic species categorized into parasite Morphospecies and its microscope image; pictures all taken at 400x. Also displays the Fecal Samples the parasites were found in; see Fecal Collection Table (Table 2) for the number of parasite Morphospecies found in each fecal sa mple. Number Identity Picture 1 Morphospecies A Giardia lamblia (protozoan) Sample 2 four axostyles, nuclei at anterior Sample 6 four flagella, nuclei at anterior Sample 20 flagella at the top of the cyst, nuclei at the bottom Sample 2 Sample 6 Sample 20 2, 6, 10, 11, 20 Total: 5 2 Morphospecies B Blastocystis hominis (protozoan cysts) 1, 2, 4, 10, 15, 16, 18, 19, 20, 22, 23 Total: 1 1 3 Morphospecies C Diphyllobothrium latum eggs (tapeworm) 2 Total: 1 4 Morphospecies D Toxocara cati (roundworm) 2 Total: 1
Giardia lamblia in White nosed Coatis Sha 14 5 Morphospecies E Entamoeba spp (roundworm cysts ) 2, 3, 5, 9, 10, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 28 Total: 21 6 Morphospecies F Ascaris lumbricoides (tapeworm egg nematode) Top iodine stain Bottom saline stain 2, 6 Total: 2 7 Morphospecies G Y east (or unidentifiable) 14 Total: 1 8 Morphospecies H U nidentifiable roundworm egg 3 Total: 1 9 Morphospecies I Coccidia 5, 21 Total: 2
Giardia lamblia in White nosed Coatis Sha 15 10 Morphospecies J Ancylostoma spp ( caninum or duodenale species ) hookworm Top egg Bottom mature worm 5 Total: 1 11 Morphospecies K U nidentifiable tapeworm egg 5 Total: 1 12 Morphospecies L 1. Isospora spp. Coccidia parasite cyst or 2. Tapeworm egg Hymenolepis spp. 4, 11 Total: 2 13 Morphospecies M both cysts 11, 20 Total: 2 14 Morphospecies N segmented cyst 13, 20 Total: 2 15 Morphospecies O mature worm 15 Total: 1
Giardia lamblia in White nosed Coatis Sha 16 16 Morphospecies P Rhabiditidae family Strongyloides spp (roundworm) Top growing worm after a few generations from emerging from egg Bottom mature worm 11, 16, 17, 18, 23, 25, 28 Total: 7 17 Morphospecies Q Trichuris vulpis or trichiura (whipworm cysts ) 4, 5, 6, 9, 11, 12, 13, 18, 21, 25 Total: 10 18 Morphospecies R cyst 5, 20 Total: 2 19 Morphospecies S Enterobius vermicularis (pinworm mature form) 22, 25, 26, 27, 28 Total: 5 20 Morphospecies T unknown cyst 28 Total: 1
Giardia lamblia in White nosed Coatis Sha 17 Table 2 : Fecal Collection Excel Sheet ( Raw Data). Includes the Fecal Sample number, date and location in which the sample was taken, Feces Diameter, Bristol Stool Chart grade, and observations regarding the visual appearance of the feces. Observations of the paras ites were also found in each fecal sample and the respective identities of each Morphospecies.
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