Black and Gold Howler Monkeys (Alouatta Caraya) As Sentinels of Ecosystem Health: Patterns of Zoonotic Protozoa Infection Relative to Degree of Human–Primate Contact

American Journal of Primatology 73:75–83 (2011)

RESEARCH ARTICLE Black and Gold Howler Monkeys (Alouatta caraya) as Sentinels of Ecosystem Health: Patterns of Zoonotic Protozoa Infection Relative to Degree of Human–Primate Contact

MARTIN M. KOWALEWSKI1, JOHANNA S. SALZER2, JOSEPH C. DEUTSCH3, MARIANA RAN˜ O1, 3 2,4Ã MARK S. KUHLENSCHMIDT , AND THOMAS R. GILLESPIE 1Estacioń Biolo´gica de Usos Mu´ltiples de Corrientes (EBCo), Museo Argentino de Ciencias Naturales-CONICET, Argentina 2Department of Environmental Studies and Program in Population Biology, Ecology, and Evolution, Emory University, Atlanta, Georgia 3Department of Pathobiology, College of Veterinary Medicine, University of Illinois, Urbana, Illinois 4Department of Environmental and Occupational Health, Rollins School of Public Health, Emory University, Atlanta, Georgia Exponential expansion of human populations and human activities within primate habitats has resulted in high potential for pathogen exchange creating challenges for biodiversity conservation and global health. Under such conditions, resilient habitat generalists such as black and gold howler monkeys (Alouatta caraya) may act as effective sentinels to overall ecosystem health and alert us to impending epidemics in the human population. To better understand this potential, we examined noninvasively collected fecal samples from black and gold howler monkeys from remote, rural, and village populations in Northern Argentina. We examined all samples (n 5 90) for the zoonotic protozoa Cryptosporidium sp. and Giardia sp. via immunofluorescent antibody (IFA) detection. All samples were negative for Cryptosporidium sp. The prevalence of Giardia sp. was significantly higher at the rural site (67%) compared with the remote forest (57%) and village (40%) sites. A lack of Cryptosporidium sp. in all samples examined suggests that this pathogen is not a natural component of the howler parasite communities at these sites and that current land-use patterns and livestock contact are not exposing Argentine howler monkeys to this pathogen. High prevalence of Giardia sp. at all sites suggests that howler monkeys may serve as a viable reservoir for Giardia. Significantly higher prevalence of Giardia sp. at the rural site, where primate–livestock contact is highest, suggests the presence of multiple Giardia clades or increased exposure to Giardia through repeated zoonotic transmission among nonhuman primates, livestock, and/or people. These results highlight the need for future research into the epidemiology, cross-species transmission ecology, and clinical consequences of Giardia and other infectious agents not only in humans and livestock, but also in the wild animals that share their environments. Am. J. Primatol. 73:75–83, 2011. r 2010 Wiley-Liss, Inc.

Key words: Cryptosporidium; disturbance ecology; Giardia;neglectedpathogens;primate;Zoonoses

INTRODUCTION primates to serve as sentinels, alerting us before pathogens reach semi-urban or urban areas. Exponential expansion of human populations The forests of Northern Argentina provide an and human activities within primate habitats has ideal locale for examining issues related to pathogen resulted in high potential for multi-directional exchange between people, nonhuman primates, and pathogen exchange creating challenges for biodiversity conservation and global health [Gillespie et al., 2008]. Associated landscape-level disturbance may Contract grant sponsor: NIH; Contract grant number: T35 2006; exacerbate pathogen transmission by affecting the Contract grant sponsors: The Wenner Gren Foundation; Post- distribution and behavior of primates [Gillespie & Ph.D. Research Grant; Contract grant number: 7622; Contract Chapman, 2006, 2008; Gillespie et al., 2005; Nunn & grant sponsor: Conservation Medicine Center of Chicago. Altizer, 2006], as well as vectors of infectious diseases ÃCorrespondence to: Thomas R. Gillespie, E510 Math and such as yellow fever, dengue, and malaria [Altizer Science Center, 400 Dowman Drive, Emory University, Atlanta, Georgia 30322. E-mail: [email protected] et al., 2006; Hay et al., 2002]. In this regard, studies of Received 28 September 2009; revised 26 December 2009; nonhuman primates with the ability to survive in revision accepted 26 December 2009 moderately disturbed habitats are of great impor- DOI 10.1002/ajp.20803 tance for understanding the dynamics of infectious Published online 18 January 2010 in Wiley Online Library (wiley disease transmission and to evaluate the capacity of onlinelibrary.com). r 2010 Wiley-Liss, Inc. 76 / Kowalewski et al.

livestock. Here, commercial agriculture (i.e., soy recent genetic differentiation among groups and a plantations) and forest exploitation have profoundly lower genetic heterozygosity, potentially due to changed the composition of forests, eliminating the limitations in dispersal opportunities [Oklander, hardiest and most economically profitable forest tree 2007]. The impacts of habitat alteration on nonhu- species [Zunino & Kowalewski, 2008]. Clearing man primate populations occur at the individual, entire forests for agriculture and establishing exotic, group, and population level. Successful mediation of fast-growing pastures are also contributing to rapid these threats requires repeated evaluation of genetic landscape change in this region. The effects of these diversity, health, stress, behavior, and diet. A study changes on wildlife populations remain largely on red colobus (Piliocolobus tephrosceles) revealed unknown. In Argentina, black and gold howler the synergistic affects of food availability, cortisol monkeys, Alouatta caraya, account for the highest levels, and parasite infections on the abundance of proportion of biomass of arboreal mammals and this species [Chapman et al., 2006]. Case studies of these monkeys often cope well with moderate various species from sites around the world will deforestation [Zunino et al., 2007]. The high degree improve our understanding of the multifactorial of phenotypic plasticity present in this species make process that anthropogenic disturbances can initiate black and gold howler monkeys a valuable species to at different levels. test and evaluate current ideas of conservation In this study we examine patterns of infection related to the dynamics of emergent and re-emergent with the zoonotic protozoa, Giardia, and Cryptospor- diseases [Kowalewski & Gillespie, 2009], since more idium, in populations of black and gold howler plastic species are expected to occupy and success- monkeys living at variable levels of contact with fully exploit a wider range of ecological conditions, humans: remote, rural, and village. These protozoa leading to greater resilience to habitat seasonality infect vertebrates, including wildlife, livestock, and and habitat modification compared with other taxa humans [Hill, 2001; McGlade et al., 2003; Volotaõ [Miner et al., 2005]. Howlers are classified as et al., 2008]. The protozoa life cycle consists of colonizers [Crockett, 1998; Crockett & Eisenberg, fecal–oral transmission of Giardia cysts or Cryptos- 1987; Eisenberg et al., 1972] due to their ability to poridium oocysts (infective stage), typically via adapt and survive in modified environments [Bicca- contaminated water and food. Within the host Marques, 2003; Clarke et al., 2002a]. This ability animal, the life cycle of the protozoa continues as to survive in variable environments including habi- the cysts or oocysts develop into trophozoites. The tats that bring them into contact with humans trophozoites infect cells within an animal host’s [Cabral et al., 2005; Estrada et al., 2006; Munõz duodenum; often leading to severe diarrhea [Thompson, et al., 2006] or increase their terrestrial travel 2004]. Giardia is cosmopolitan and constitutes a [Kowalewski et al., 1995; Pozo-Montuy & Serio- recognized waterborne infection for humans and Silva, 2006; Young, 1981], make howlers an excellent animals, responsible for well over one billion cases of model to study the dynamics of infectious disease human diarrheal disease annually [Crompton, 2000; transmission among wild primates, humans, and Teodorovic et al., 2007; WHO/UNICEF, 2000]. domestic animals. Giardiasis is now considered by the World Health Although black and gold howlers, much like Organization to be a ‘‘neglected disease,’’ due to its other species of Alouatta, seem to be resilient to ubiquity in equatorial latitudes, its chronic detri- variable levels of habitat degradation in terms of mental health effects (especially in children), its population numbers [Estrada et al., 2006; Zunino propensity to infect economically disadvantaged et al., 2007], a finer-scale analysis reveals a different populations, and the paradoxical availability of scenario. As in colobine species [Gillespie & Chapman, relatively inexpensive and effective pharmaceuticals 2006, 2008], howler populations living in fragmented for its treatment [Savioli et al., 2006]. In Latin forests seem to be carrying a more diverse commu- America almost 15% of the rural population is nity of parasites [Kowalewski & Gillespie, unpub- infected with Giardia [Atıás, 1999]. In Argentina, lished data; Santa Cruz et al., 2000]. Oklander [2007] two human studies showed a prevalence of 10% in revealed another trend in howler populations living urban areas, 34% in small towns (same as villages in fragmented forests in Northern Argentina (the considered in our study) [Gamboa et al., 2003], and same study site of this article—EBCo-, see descrip- 4% in rural areas [Minvielle et al., 2004]. In animals tion in methods). Oklander [2007] used molecular from this region the reported prevalence of Giardia techniques [13 polymorphic microsatellites devel- is 7.5% in cattle, 18.6% in dogs, and 3.9% in rodents oped by Oklander et al., 2007] to compare levels of [Pezzani et al., 2003]. These parasites have been homozygosity in a population living in a continuous reported in other nonhuman primates including forest and a population living in a fragmented forest mountain gorillas (Gorilla gorilla berengei) [Graczyk (same rural population studied in this article) by et al., 2002], black howler monkeys (Alouatta pigra) estimating gene flow with Fst values. In contrast to [Vitazkova & Wade, 2006], brown howling monkey groups living in continuous forests (our IB site, see (Alouatta guariba) [Cabral et al., 2005; Volotaõ et al., methods), groups living in fragments presented a 2008], red colobus (Pilocolobus tephrosceles) [Salzer

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et al., 2007] and red-tailed guenons (Cercopithecus approximately 30 ha) covering a total area of 3,000 ha ascanius) [Salzer et al., 2007]. As evidenced from the [see Zunino et al., 2007 for details]. This site is under examples above, giardiasis is often endemic in continuous deforestation due to selective logging and human populations; however, the frequency and clearing of fragments for cattle ranches and howlers pathogenicity of this agent remains largely unknown have the potential of close contact with cattle, other in wildlife [Appelbee et al., 2005; Levy et al., 1998]. domestic animals, and humans when accessing The main symptoms of giardiasis in humans are shared water sources and during terrestrial travel. diarrhea, abdominal pain, nausea, and vomiting Terrestrial travel is typical of other howler popula- [Acha & Szyfres, 2003]. The disease is generally tions living in similar conditions [Clarke et al., described as an acute diarrheal illness that may last 2002b; Glander, 1992; Horwich & Lyon, 1998; up to six weeks although chronic patients are also Pozo-Montuy & Serio-Silva, 2006]. Human houses found, especially in cases of impaired immunocom- are distributed uniformly across this rural site. We petence [Chester et al., 1985]. also sampled howlers from two village sites, Cerrito town (C) near IB (there are two groups of howlers METHODS living in this town of 1,500 human inhabitants) and SC next to the EBCO with almost 4,000 human Study Sites inhabitants (there are 7–8 groups living in close This study was conducted in four sites that proximity with family houses whose tree crowns are differ in their degree of human use: Isla Brasilera contiguous with trees in the forest fragments). In (IB) (271200S, 581400W), Estacioń Biolo´gica Cor- summary, the four areas were classified into three rientes-Biological Field Station Corrientes- (EBCO) habitat types: remote (IB), rural (EBCO), and village (271300S, 581410W), Cerrito (C) (271170S, 581370W), (C and SC). The climate of all sites is subtropical and San Cayetano (SC) (271340S, 581420W) (Fig. 1). with an average annual temperature of 21.61C and IB is an island characterized by a continuous flooded an annual average of rainfall of 1,200 mm [Rumiz forest where howler groups exhibit extensive home et al., 1986]. Rains increase slightly towards the range overlap, but little to no human contact. The spring–summer (September to December). island has an area of 290 ha and we have identified between 32–35 groups. IB is 20 km north of EBCO (Fig. 1), a rural site characterized by semi-deciduous Study Population forest surrounded by a matrix of grassland. At We collected 90 fecal samples from different EBCO, in general there is one group of howlers per individuals during March 2008 (fall season): 30 forest fragment. We have identified 34 groups of samples from the village sites (C and SC), 30 samples howlers in 24 forest fragments (area from 3 ha to from the rural site (EBCO) and 30 samples from the

Fig. 1. Location of study areas of black and gold howler monkeys (Alouatta caraya) in northern Argentina: San Cayetano (SC) and Cerrito (C) were considered village sites, Isla Brasilera (IB) a remote site, and EBCO a rural site.

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remote site (IB). Each sample set consisted of 30 samples and included 10 adult males, 10 adult females, 4 adult females with lactating infants, and approximately 3 juvenile males and 3 juvenile females (the number of juveniles varied across groups). These samples were collected from different groups of howlers in each site.

Sample Analysis Fecal samples were collected immediately after defecation in (March 2008) and preserved in 10% neutral buffered formalin [Gillespie, 2006]. When collected, fecal consistency was characterized as liquid, soft, medium, or solid. We used a Merifluor Cryptosporidium/Giardia Direct Immunofluorescent Detection Kit (Meridian Bioscience Inc, Cincinnati, Fig. 2. Giardia sp. cysts/gram in black and gold howler monkey Ohio) to detect both parasites of interest—Crysto- (Alouatta caraya) feces from remote, rural, and urban sites in poridium oocysts and Giardia cysts based on mono- northern Argentina. clonal antibodies [Johnston et al., 2003]. The analysis was run in July 2008. Fecal samples were differences were found between rural and village scored both for the presence or absence of the areas (w2 test, df 5 1, Po0.03). No fecal samples were pathogens as well as quantification of Cryptospor- scored as liquid. From a total of 49 positive samples, idium oocysts and Giardia cysts in feces. Fecal 15 (31%) were characterized as soft, 27 (55%) as samples were concentrated and then resuspended medium and 7 (14%) as solid. These results suggest in 1 g/1 mL suspensions. Counts were calculated by that the consistency of fecal material is independent analyzing 10 mL of the 1 g/mL solution of concen- of the infected state of the individuals (w2 test, df 5 2, trated feces from each sample. Oocysts or cysts were Po0.05). then quantified by counting total numbers in 150 We also compared cysts per gram of fecal microscope fields (400 magnification) and extra- material, providing an index of intensity of shedding polating results to the entire sample. This method and environmental contamination with Giardia. was validated by spiking negative fecal samples with Although the mean value of cysts per gram was known numbers of Cryptosporidium sp. oocysts. highest in remote areas (250.9, range 5 4–1,064), followed by village areas (194.7, range 5 1–790) and Statistical Analyses rural areas (113.5, range 5 1–845); there were no Differences in the frequency and prevalence of significant differences in the relative number of cysts infections across sites and differences in frequencies per sample across areas (Kruskall Wallis ANOVA, of fecal consistency were compared using w2 tests. We H¼2ðN¼49Þ ¼3, P40.05) (Fig. 2). used Kruskall Wallis ANOVA to explore differences of intensity of infection across sites. We considered a DISCUSSION criterion for significance for all statistical tests at Po0.05. This research complied with the American Our results demonstrate that Giardia sp. is Journal of Primatology and University of Illinois present and prevalent in wild black and gold howlers guidelines for the ethical treatment of primates at all points along a gradient of anthropogenic (IACUC protocol 06207) and the laws of Argentina. disturbance. In these same populations, there is no evidence of infection with Cryptosporidium sp. A lack of Cryptosporidium sp. in all samples examined RESULTS suggests this pathogen is not a natural component All samples were negative for Cryptosporidium of the howler parasite communities at these sites and sp. Seventeen of 30 samples (57%) in the remote site, that current land-use patterns are not exposing 20 of 30 samples (67%) in the rural area, and 12 of 30 Argentine howler monkeys to this pathogen. Cryp- samples (40%) in the village areas were positive for tosporidium oocysts were found in people in urban Giardia sp. When the remote, rural, and village sites and semi-urban populations close to our study sites were compared collectively, the prevalence of Giardia [Ledesma et al., 2006], but it was never reported for sp. infection in black and gold howler monkeys was the rural area considered in this article [Borda et al., not significantly related to the location of sampling 1996]. (w2 test, df 5 2, P>0.05). When sampling locations High prevalence of Giardia sp. at all sites were compared individually (remote vs. village, regardless of human–primate contact rates suggest rural vs. village, and remote vs. rural), significant that howler monkeys may serve as a viable reservoir

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for Giardia sp. Considered among the most common There are several reasons to expect higher rates human intestinal protozoa, Giardia ranges in clinical of contact between howlers and cattle, a common presentation from asymptomatic to highly patho- source of protozoal zoonotics, in this rural site. First, genic, causing chronic malabsorptive diarrhea in cattle routinely enter forest fragments inhabited by some patients [Thompson, 2004]. Both host factors howlers looking for shade and young pastures, (e.g., nutrition, immunity, co-infection with other opening trails and defecating along them. Second, agents) and pathogen factors (e.g., strain, infectious howlers drink water from creeks where cattle drink dose) are thought to contribute to the severity of and defecate [Kowalewski & Ranõ, personal observa- clinical disease [Thompson, 2000]. In our study, we tion]. Finally, increased levels of deforestation did not characterize any howler monkey fecal decrease the size of these fragments and obligate samples as liquid, however; we did find that infected howlers to descend to cross from fragment to animals had more fecal samples scored as soft and/or fragment looking for food resources [Zunino et al., medium than noninfected animals. The lack of a 2007] increasing the likelihood of contact with strong measurable association between infection and parasites on the ground and in small water bodies. an acute gastrointestinal symptom (i.e., diarrhea) in As in the case of the rural site, we also found our study population was surprising, but it is high prevalence of Giardia in howler populations in concordant with recent studies of humans. Although the two other sites (remote and village). Giardia had Giardia is associated with clinical disease in devel- a prevalence of 57% in the remote site. Although this oped economies, evidence is nevertheless mounting site lacks permanent human settlements, there are that it may be commensal in rural settings where occasional visitors from neighboring towns that people may normally be exposed to diverse parasite bring a limited number (10–20) of cattle onto the communities from an early age. For example, Cordoń island, which drink from the same lagoons that the et al. [2008] found Giardia at 28.1% prevalence in monkeys use as a water source [Bravo & Sallenave, diarrheic Peruvian children, as well as in 19.5% of 2003; Kowalewski, 2007]. Giardia duodenalis was nondiarrheic children, emphasizing the importance reported in brown howling monkeys (Alouatta of asymptomatic patients in Giardia transmission guariba), as the result of contact of these monkeys where hygiene and sanitation are poor [Cordoń et al., with other wildlife, such as opossums, which live in a 2008]. Other studies in India [Traub et al., 2003], nearby forest fragment and circulate near human Ethiopia [Ayalew et al., 2008], Bangladesh [Dib et al., dwellings [Muller et al., 2000; Volotaõ et al., 2007] or 2008], and Peru [Hollm-Delgado et al., 2008] have contact with humans such as fishermen, poachers, or found similarly weak or nonexistent associations researchers [Volotaõ et al., 2007; see also Vitazkova between G. duodenalis infection status and gastro- & Wade, 2007]. Similarly, intensified gorilla–human intestinal symptoms, suggesting that G. duodenalis interaction has been suggested to enhance transmis- may coexist benignly with human hosts under sion of anthropogenic pathogens including Giardia conditions where acquired immunity to the pathogen [see Graczyk et al., 2001a,b, 2002; Nizeyi et al., can develop. Lack of early exposure and acquired 1999]. immunity to G. duodenalis may, in fact, account for Although the village site in our study revealed the role of the pathogen as a cause of sporadic the lowest prevalence of Giardia (40%), this value is diarrheal outbreaks in developed countries [Istre high relative to Giardia prevalence in other wild et al., 1984], as well as a major cause of traveler’s primate populations examined. A study in 1983 of diarrhea for people from developed countries who visit children from the public school at this site (SC) endemic areas [Reinthaler et al., 1998; Shlim et al., showed a 29% (n 5 60) prevalence of Giardia lambia 1999]. The interplay between clinical symptoms and (5 Giardia duodenalis) infection [Borda et al., 1996]. Giardia infections in wildlife are far less known. We The interactive affects of poverty, low levels of hope to explore the association between symptom and education, unsanitary conditions, and increased infection in black and gold howler monkeys and other levels of deforestation experienced at this site may wildlife species in more detail in future work. result in increased contact between wildlife and Giardia is also notable for zoonotic transmission humans, producing unexpected results in the trans- [Thompson, 2004]. The protozoan’s propensity to mission patterns of infectious diseases. cross species barriers results from the host non- Vitazkova and Wade [2007] suggested a positive specificity of the trophozoite as well as from the high relationship between primate density and prevalence infectivity and environmental stability of the Giar- of Giardia sp. in a population of Alouatta pigra dia cyst [Smith & Nichols, 2006]. The fact that during the dry season in Belize and Mexico. In our Giardia sp. prevalence was significantly higher at study, we find the opposite trend with highest the rural site in our study, where monkeys have the prevalence at the site with the lowest primate highest contact rates with cattle and other domestic density (1.04 howlers/ha for the rural site) and animals, suggests that repeated zoonotic transmis- lowest prevalence at the site with the highest sion may amplify baseline rates of infection [Zunino primate density (3.25 howlers/ha for the remote site) & Kowalewski, 2008]. [Kowalewski & Zunino, 2004; Zunino et al., 2007].

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This pattern suggests that other factors associated in poverty-stricken communities where health sys- with anthropogenic disturbance such as physical tems are inadequate. Our study is an illustration of characteristics of sites (humidity, water availability), how these exploratory evaluations may benefit these history of both parasites and hosts in sites, variation efforts. Establishment of long-term study sites allow in the use of space, and stress may trump host researchers to catalog rare events and build knowl- density in driving patterns of infection with directly edge and collaborations over time to better under- transmitted pathogens. stand the dynamics of infectious diseases and to Black and gold howlers living in fragmented potentially prevent future outbreaks. For example, areas have shown a higher diversity of gastrointest- data presented in this article are part of a long-term inal parasites [Kowalewski & Gillespie, unpublished and ongoing study of ecosystem health in northern data] potentially due to an increase in contact with Argentina, which includes an evaluation of several cattle, domestic animals, and humans. In addition, a pathogens in humans, domesticated animals and study carried out at the same rural site has shown nonhuman primates among other wild mammals. In that howlers have higher levels of homozygosity than addition to incorporating government institutions in areas remote from human occupations with and public universities, our data collection and continuous forests [Oklander, 2007; Oklander et al., sampling design allows the comparison of our results 2007]. Collectively, this suggests that howlers are with similar projects carried out using standardized viable sentinels for overall ecosystem health. For methods around the globe [Gillespie, 2006; Gillespie example, in 2007, an outbreak of yellow fever et al., 2008]. Impoverished people with limited access decimated the population of A. caraya and A. guariba to basic hygienic services or adequate health systems in the Province of Misiones (Argentina) located are often most susceptible to emerging diseases. north of the field sites of the current study [Agostini These same people and wildlife that live in surround- et al., 2008; Holzmann, personal communication]. ing habitats are often the most affected by the This outbreak also affected humans in the area. In negative consequences of large-scale habitat modifi- 2008, one year after these reports were made, we cations, despite being rarely responsible for such found dead howlers (A. caraya) in the same province habitat modification [Alarcoń-Chaíres, 2006]. Local (Corrientes) that likely succumbed to yellow fever and national governments must invest resources in near our study site. Unfortunately, supplies were not development for such communities in terms of available to allow for diagnostic confirmation. The education, health, and conservation programs and yellow fever wave is progressing to the South of hold large local landowners and international cor- Argentina. Howler deaths constitute an important porations that produce the highest impact on wildlife signal of an impending public health threat that can through the establishment of pastures and large- alert officials of the need to intervene to block scale plantations of soy, oil palm, rice, and other conversion of a sylvatic (i.e., forest cycle) into an global agricultural commodities accountable for their urban cycle of this deadly human disease. Black and impacts on human, animal, and ecosystem health gold howlers are effective sentinels of yellow fever, [De la Cruz, 2004; Martıńez Alier, 2005]. and ongoing study of their populations may help us Researchers can play an important role in to prevent and better understand the yellow fever conservation and development that extends far transmission dynamics. Beyond the case of yellow beyond the dissemination of results through inter- fever, resilient habitat generalists such as the black national scientific journals. They can expand their and gold howlers can act as effective sentinels of capacity by assembling multidisciplinary teams for overall ecosystem health and alert us to impending conservation and health applications, establishing epidemics in human populations. Consequently, we education programs that incorporate local people and view the results of this study as indicating a strong researchers in areas of study, and conveying infor- need for future research into the epidemiology, cross- mation to local communities through radio, news- species transmission ecology, and clinical conse- paper, and community meetings. Most importantly, quences of Giardia and other infectious agents not researchers can help colleagues and students in only in humans and livestock, but also in the wild developing countries to obtain training and funding animals that share their environments. for ongoing and proposed research. Our current knowledge regarding wild primate pathogens remains minimal and skewed toward Recommendations regions of long-term primate research. Fortunately, Successful outcomes involving emerging and integration of standardized empirical data collection, remerging infectious diseases require a proactive state-of-the-art diagnostics, and the comparative approach, ideally avoiding introduction of infectious approach offers the opportunity to create a baseline agents into human, livestock, and threatened for patterns of infection in wild primate populations; wildlife populations altogether. Consequently, sur- to better understand the role of disease in primate veillance efforts need to be strongest in areas of ecology, behavior, and evolution; and to examine how increasing contact between humans and wildlife and anthropogenic effects alter the zoonotic potential of

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