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Cysts and trophozoites of vestibuliferid ciliates and larvae of Strongyloides were found in fecal samples from captive orangutans Pongo pygmaeus and P. abelii from Czech and Slovak zoological gardens. As comparative material, ciliates from semi-captive mandrills Mandrillus sphinx from Gabon were included in the study. Phylogenetic analysis of the detected vestibuliferid ciliates using ITS1-5.8s-rRNA-ITS2 and partial 18S rDNA revealed that the ciliates from orangutans are conspecific with Balantioides coli lineage A, while the ciliates from mandrills clustered with Buxtonella-like ciliates from other primates. Morphological examination of the cysts and trophozoites using light microscopy did not reveal differences robust enough to identify the genera of the ciliates. Phylogenetic analysis of detected L1 larvae of Strongyloides using partial cox1 revealed Strongyloides stercoralis clustering within the cox1 lineage A infecting dogs, humas and other primates. The sequences of 18S rDNA support these results. As both B. coli and S. stercoralis are zoonotic parasites and the conditions in captive and semi-captive settings may facilitate transmission to humans, prophylactic measures should reflect the findings.

Rapid increases in human populations and environmental changes of past decades have led to intensified contact with wildlife and significantly contributed to pathogen transmission in both directions, especially between humans and non-human primates, whose close phylogenetic relationship facilitates cross-infection. Using high-throughput sequencing, we studied strongylid communities in sympatric western lowland gorillas, central chimpanzees and humans co-occurring in an unprotected area in the northern periphery of the Dja Faunal Reserve, Cameroon. We identified 65 strongylid ITS-2 amplicon sequencing variants (ASVs) in humans and great apes. Great apes exhibited higher strongylid diversity than humans. Necator and Oesophagostomum were the most prevalent genera, and we commonly observed mixed infections of more than one strongylid species. Human strongylid nematodes were dominated by the human hookworm N. americanus, while great apes were mainly infected with N. gorillae, O. stephanostomum and trichostrongylids. We were also able to detect rare strongylid taxa (such as Ancylostoma and Ternidens). We detected eight ASVs shared between humans and great apes (four N. americanus variants, two N. gorillae variants, one O. stephanostomum type I and one Trichostrongylus sp. type II variant). Our results show that knowledge of strongylid communities in primates, including humans, is still limited. Sharing the same habitat, especially outside protected areas (where access to the forest is not restricted), can enable mutual exchange of parasites and can even override host phylogeny or conserved patterns.

Zoonotic pathogen transmission is considered a leading threat to the survival of non-human primates and public health in shared landscapes. Giardia spp., Cryptosporidium spp. and Microsporidia are unicellular parasites spread by the fecal-oral route by environmentally resistant stages and can infect humans, livestock, and wildlife including non-human primates. Using immunoassay diagnostic kits and amplification/sequencing of the region of the triosephosphate isomerase, small ribosomal subunit rRNA and the internal transcribed spacer genes, we investigated Giardia, Cryptosporidium, and microsporidia infections, respectively, among humans, domesticated animals (livestock, poultry, and dogs), and wild nonhuman primates (eastern chimpanzees and black and white colobus monkeys) in Bulindi, Uganda, an area of remarkably high human–animal contact and spatial overlap. We analyzed 137 fecal samples and revealed the presence of G. intestinalis assemblage B in two human isolates, G. intestinalis assemblage E in one cow isolate, and Encephalitozoon cuniculi genotype II in two humans and one goat isolate. None of the chimpanzee and colobus monkey samples were positive for any of the screened parasites. Regular distribution of antiparasitic treatment in both humans and domestic animals in Bulindi could have reduced the occurrence of the screened parasites and decreased potential circulation of these pathogens among host species.

The close phylogenetic relationship between humans and nonhuman primates (NHPs) can result in a high potential for pathogen exchange. In recent decades, NHP and human interactions have become more frequent due to increasing habitat encroachment and ecotourism. Strongylid communities, which include members of several genera, are typically found in NHPs. Using optimized high-throughput sequencing for strain-level identification of primate strongylids, we studied the structure of strongylid communities in NHPs and humans co-habiting a tropical forest ecosystem in the Central African Republic. General taxonomic assignment of 85 ITS-2 haplotypes indicated that the studied primates harbour at least nine genera of strongylid nematodes, with Oesophagostomum and Necator being the most prevalent. We detected both host-specific and shared strongylid haplotypes. Skin-penetrating Necator gorillaehaplotypes were shared between humans and gorillas but Necator americanus were much more restricted to humans. Strongylid communities of local hunter-gatherers employed as trackers were more similar to those of gorillas compared to their relatives, who spent more time in villages. This was due to lower abundance of human-origin N. americanus in both gorillas and trackers. Habituated gorillas or those under habituation did not show larger overlap of strongylids with humans compared to unhabituated. We concluded that the occurrence of the human-specific strongylids in gorillas does not increase with direct contact between gorillas and humans due to the habituation. Overall, our results indicate that the degree of habitat sharing between hosts, together with mode of parasite transmission, are important factors for parasite spillover among primates.

We investigated intestinal trichomonads in western lowland gorillas, central chimpanzees and humans cohabiting the forest ecosystem of Dzanga-Sangha Protected Area in Central African Republic, using the internal transcribed spacer (ITS) region and SSU rRNA gene sequences. Trichomonads belonging to the genus Tetratrichomonas were detected in 23% of the faecal samples and in all host species. Different hosts were infected with different genotypes of Tetratrichomonas. In chimpanzees, we detected tetratrichomonads from ‘novel lineage 2’, which was previously reported mostly in captive and wild chimpanzees. In gorillas, we found two different genotypes of Tetratrichomonas. The ITS region sequences of the more frequent genotype were identical to the sequence found in a faecal sample of a wild western lowland gorilla from Cameroon. Sequences of the second genotype from gorillas were almost identical to sequences previously obtained from an anorexic French woman. We provide the first report of the presence of intestinal tetratrichomonads in asymptomatic, apparently healthy humans. Human tetratrichomonads belonged to the lineage 7, which was previously reported in domestic and wild pigs and a domestic horse. Our findings suggest that the ecology and spatial overlap among hominids in the tropical forest ecosystem has not resulted in exchange of intestinal trichomonads among these hosts.

Wildlife ecotourism can offer a source of revenue which benefits local development and conservation simultaneously. However, habituation of wildlife for ecotourism can cause long-term elevation of glucocorticoid hormones, which may suppress immune function and increase an animal’s vulnerability to disease. We have previously shown that western lowland gorillas (Gorilla gorilla gorilla) undergoing habituation in Dzanga-Sangha Protected Areas, Central African Republic, have higher fecal glucocorticoid metabolite (FGCM) levels than both habituated and unhabituated gorillas. Here, we tested the relationship between FGCM levels and strongylid infections in the same gorillas. If high FGCM levels suppress the immune system, we predicted that FGCM levels will be positively associated with strongylid egg counts and that gorillas undergoing habituation will have the highest strongylid egg counts, relative to both habituated and unhabituated gorillas. We collected fecal samples over 12 months in two habituated gorilla groups, one group undergoing habituation and completely unhabituated gorillas. We established FGCM levels and fecal egg counts of Necator/Oesophagostomum spp. and Mammomonogamus sp. Controlling for seasonal variation and age-sex category in strongylid infections we found no significant relationship between FGCMs and Nectator/Oesophagostomum spp. or Mammomonogamus sp. egg counts in a within group comparison in either a habituated group or a group undergoing habituation. However, across groups, egg counts of Nectator/Oesophagostomum spp. were lowest in unhabituated animals and highest in the group undergoing habituation, matching the differences in FGCM levels among these gorilla groups. Our findings partially support the hypothesis that elevated glucocorticoids reduce a host’s ability to control the extent of parasitic infections, and show the importance of non-invasive monitoring of endocrine function and parasite infection in individuals exposed to human pressure including habituation process and ecotourism.

Conservation efforts have led to the recovery of the endangered mountain gorilla populations. Due to their limited potential for spatial expansion, population densities increased, which may alter the epidemiology of infectious diseases. Recently, clinical gastrointestinal illnesses linked to helminth infections have been recorded in both gorilla populations. To understand drivers and patterns of helminth infections we quantified strongylid and tapeworm infections across both Virunga Massif and Bwindi populations using fecal egg counts. We assessed the impact of age, sex, group size, season and spatial differences used as a proxy, which reflects observed variation in the occurrence of gastrointestinal problems, vegetation types, gorilla subpopulation growth and associated social structure on helminth infections. We revealed striking geographic differences in strongylid infections with higher egg counts mostly in areas with high occurrences of gastrointestinal disease. Increased helminth egg counts were also associated with decreasing group size in some areas. Observed spatial differences may reflect mutual effects of variations in subpopulation growth rates, gorilla social structure, and vegetation associated with altitude across mountain gorilla habitat. Helminth infection intensities in Virunga gorillas were lowest in the youngest and the oldest animals. Elucidating parasite infection patterns of endangered species with low genetic diversity is crucial for their conservation management.

Blastocystis is the most commonly found intestinal protist in the world. Accurate detection and differentiation of Blastocystis including its subtypes (arguably species) are essential to understand its epidemiology and role in human health. We compared (i) the sensitivity of conventional PCR (cPCR) and qPCR in a set of 288 DNA samples obtained from stool samples of gut-healthy individuals, and (ii) subtype diversity as detected by next-generation sequencing (NGS) versus Sanger sequencing. Real-time PCR resulted in more positive samples than cPCR, revealing high fecal load of Blastocystis based on the quantification curve in most samples. In subtype detection, NGS was largely in agreement with Sanger sequencing but showed higher sensitivity for mixed subtype colonization within one host. This fact together with use of the combination of qPCR and NGS and obtaining information on the fecal protist load will be beneficial for epidemiological and surveillance studies.Comparaison des approches de diagnostic moléculaire pour la détection et la différenciation du protiste intestinal Blastocystis sp. chez l’homme.Blastocystis est le protiste intestinal le plus répandu dans le monde. La détection et la différenciation précises de Blastocystis, y compris ses sous-types (sans doute des espèces), sont essentielles pour comprendre son épidémiologie et son rôle dans la santé humaine. Nous avons comparé (i) la sensibilité de la PCR conventionnelle (cPCR) et de la qPCR dans un ensemble de 288 échantillons d’ADN obtenus à partir d’échantillons de selles d’individus en bonne santé intestinale et (ii) la diversité des sous-types détectée par le séquençage de nouvelle génération (NGS) par rapport au séquençage Sanger. La PCR en temps réel a donné plus d’échantillons positifs que la cPCR, révélant une charge fécale élevée de Blastocystis sur la base de la courbe de quantification dans la plupart des échantillons. Dans la détection des sous-types, le NGS était largement en accord avec le séquençage de Sanger mais a montré une sensibilité plus élevée pour la colonisation de sous-types mixtes au sein d’un hôte. Ce fait, associé à l’utilisation de la combinaison de qPCR et de NGS et à l’obtention d’informations sur la charge fécale de protistes, sera bénéfique pour les études épidémiologiques et de surveillance.

International audience; Gastrointestinal nematodes are known to be one of the most economically important parasites in livestock production. In order to test whether fractal analysis of behaviour can be used as a diagnostic tool for detection of infected animals, we investigated fractal patterns in the behavioural activity of domestic sheep (Ovis aries) in relation to strongylid infection. Temporal dynamics in activity patterns of 20 sheep were recorded at high resolutions using tri-axial accelerometer loggers attached to the neck of naturally infected subjects. We measured fractal dynamics in the resultant acceleration time series, divided into periods of activity and inactivity, using several fractal methods and tested the prediction that temporal complexity in the activity patterns of infected control sheep and experimentally dewormed sheep should differ. Detrended fluctuation analysis (DFA) revealed that sheep behaviour sequences were characterized by long-range correlations, meaning that acceleration fluctuations are not random but depend on long-term activity events. Generalized linear mixed models built to test for the effect of deworming on fractal estimates showed that the temporal organization of sheep activity varies with the status of strongylid infection. Our results indicate that sheep treated with anthelmintics exhibited a higher complexity in their activity sequences than parasitized sheep, suggesting that organizational patterns of their behaviour change with gastrointestinal parasite infection. Thus, we provide evidence for the potential utility of fractal methods in behavioural welfare monitoring.

The tick subgenus Afrixodes Morel 1966 (Ixodidae) is distributed in sub-Saharan Africa. It consists of about 60 species which are described based on a few specimens and/or only some developmental stages, mostly adults. Because of this, the diagnosis of the African species of Ixodes is in many aspects complicated. Ixodes rasus Neumann 1899 is a common species, widely distributed in tropical and subtropical moist broadleaved forests in central Africa, whose morphology remains poorly described. We redescribe the adults of the species and provide molecular data for 16S rDNA based on three questing females and one male collected in the Central African Republic. Both sexes have an anal groove rounded and closed. The female has syncoxae on coxae I to III, a dental formula of 2/2, and long, curved auriculae. The male has moderate syncoxae on coxae I–III, coxae I–II carry one single internal spur each: coxae III and IV unarmed. The cornua are absent in the male, and the auriculae are small lobes. The male pregenital plate is longer than broad, with an anterior margin widely rounded; the median plate is large and clearly divergent posteriorly, with adanal plates fused posteriorly to the anus, and a dental formula of 4/4 (one row of 5/5). The molecular features of 16S rDNA place I. rasus near I. aulacodi (the only other Afrixodes species for which 16S rDNA is available) and near I. turdus and I. frontalis; two species of ticks of the Palearctic region, which commonly infest birds. The paucity of data regarding the subgenus Afrixodes makes it necessary to provide reliable and comparable re-descriptions of several species, as well as description of the immature stages for many of the currently known taxa.

Objectively measuring the effects of parasitism on animal health is challenging, especially in the wild. Analyses of behavioural organization are increasingly used for this purpose, to identify animals in pathological or otherwise challenged states. Here, we investigated the possible impact of gastrointestinal helminth infection on the behaviour of wild chimpanzees, Pan troglodytes schweinfurthii, by applying fractal analysis to their feeding patterns. We predicted that higher intensity of strongylid infection should be associated with altered organizational complexity in temporal sequences of behaviour. We observed 15 habituated male chimpanzees in Kalinzu Reserve Forest, Uganda, and collected behavioural time series via focal animal sampling. We quantified the number of strongylid eggs per gram of faecal sediment using a modified simple sedimentation method to estimate the intensity of infection with strongylid nematodes. We used detrended fluctuation analysis (DFA) to explore long-range dependence in binary sequences of feeding behaviour as an index of organizational complexity along a stochastic-deterministic gradient. We then built several generalized linear mixed models to examine the relationship between behavioural organization and strongylid infection. Our results indicate that chimpanzee feeding sequences are long-range dependent and antipersistent, i.e. short bouts tended to be followed by long bouts and vice versa. Furthermore, the complexity of chimpanzee feeding sequences and the intensity of infection with strongylid nematodes were positively related: individuals with more intense infections exhibited more stochastic feeding sequences. In contrast, more conventional analyses did not reveal any relationship between parasitism and chimpanzee behaviour, nor did a survival analysis find variation in the probability of switching between behaviour states across chimpanzees with varying infection phenotypes. This work suggests that strongylid nematodes do pose a challenge for wild chimpanzees, manifest as altered organizational complexity in behaviour sequences, and provides further evidence that fractal analyses can have a valuable role in animal health monitoring.

Human malaria parasites have rarely been reported from free-ranging great apes. Our study confirms the presence of the human malaria parasite Plasmodium ovale wallikeri in western lowland gorillas and humans in Dzanga Sangha Protected Areas, Central African Republic, and discusses implications for malaria epidemiology.

In our most recent study, we found that in Tanzania infection with Treponema pallidum (TP) subsp. pertenue (TPE) is present in four different monkey species. In order to gain information on the diversity and epidemiological spread of the infection in Tanzanian nonhuman primates (NHP), we identified two suitable candidate genes for multi-locus sequence typing (MLST). We demonstrate the functionality of the MLST system in invasively and non-invasively collected samples. While we were not able to demonstrate frequent interspecies transmission of TPE in Tanzanian monkeys, our results show a clustering of TPE strains according to geography and not host species, which is suggestive for rare transmission events between different NHP species. In addition to the geographic stability, we describe the relative temporal stability of the strains infecting NHPs and identified multi-strain infection. Differences between TPE strains of NHP and human origin are highlighted. Our results show that antibiotic resistance does not occur in Tanzanian TPE strains of NHP origin.

Colonization by the benign tapeworm, Hymenolepis diminuta, has been associated with a reduction in intestinal inflammation and changes in bacterial microbiota. However, the role of microbiota in the tapeworm anti-inflammatory effect is not yet clear, and the aim of this study was to determine whether disruption of the microflora during worm colonization can affect the course of intestinal inflammation. We added a phase for disrupting the intestinal microbiota using antibiotics to the experimental design for which we previously demonstrated the protective effect of H. diminuta. We monitored the immunological markers, clinical parameters, bacterial microbiota, and histological changes in the colon of rats. After a combination of colonization, antibiotics, and colitis induction, we had four differently affected experimental groups. We observed a different course of the immune response in each group, but no protective effect was found. Rats treated with colonization and antibiotics showed a strong induction of the Th2 response as well as a significant change in microbial diversity. The microbial results also revealed differences in the richness and abundance of some bacterial taxa, influenced by various factors. Our data suggest that interactions between the tapeworm and bacteria may have a major impact on its protective effect.

The mammalian gastrointestinal (GI) microbiome, which plays indispensable roles in host nutrition and health, is affected by numerous intrinsic and extrinsic factors. Among them, antibiotic (ATB) treatment is reported to have a significant effect on GI microbiome composition in humans and other animals. However, the impact of ATBs on the GI microbiome of free-ranging or even captive great apes remains poorly characterized. Here, we investigated the effect of cephalosporin treatment (delivered by intramuscular dart injection during a serious respiratory outbreak) on the GI microbiome of a wild habituated group of western lowland gorillas (Gorilla gorilla gorilla) in the Dzanga Sangha Protected Areas, Central African Republic. We examined 36 fecal samples from eight individuals, including samples before and after ATB treatment, and characterized the GI microbiome composition using Illumina-MiSeq sequencing of the bacterial 16S rRNA gene. The GI microbial profiles of samples from the same individuals before and after ATB administration indicate that the ATB treatment impacts GI microbiome stability and the relative abundance of particular bacterial taxa within the colonic ecosystem of wild gorillas. We observed a statistically significant increase in Firmicutes and a decrease in Bacteroidetes levels after ATB treatment. We found disruption of the fibrolytic community linked with a decrease of Ruminoccocus levels as a result of ATB treatment. Nevertheless, the nature of the changes observed after ATB treatment differs among gorillas and thus is dependent on the individual host. This study has important implications for ecology, management, and conservation of wild primates.

SUMMARYTo address the molecular diversity and occurrence of pathogenic species of the genusEntamoebaspp. in wild non-human primates (NHP) we conducted molecular-phylogenetic analyses onEntamoebafrom wild chimpanzees living in the Issa Valley, Tanzania. We compared the sensitivity of molecular [using a genus-specific polymerase chain reaction (PCR)] and coproscopic detection (merthiolate-iodine-formaldehyde concentration) ofEntamoebaspp. We identifiedEntamoebaspp. in 72 chimpanzee fecal samples (79%) subjected to species-specific PCRs for sixEntamoebaspecies/groups (Entamoeba histolytica, Entamoeba nuttalli, Entamoeba dispar, Entamoeba moshkovskii, Entamoeba coliandEntamoeba poleckiST2). We recorded threeEntamoebaspecies:E. coli(47%),E. dispar(16%),Entamoeba hartmanni(51%). Coproscopically, we could only distinguish the cysts of complexE. histolytica/dispar/moshkovskii/nuttalliandE. coli. Molecular prevalence of entamoebas was higher than the prevalence based on the coproscopic examination. Our molecular phylogenies showed that sequences ofE. disparandE. colifrom Issa chimpanzees are closely related to sequences from humans and other NHP from GenBank. The results showed that wild chimpanzees harbourEntamoebaspecies similar to those occurring in humans; however, no pathogenic species were detected. Molecular-phylogenetic methods are critical to improve diagnostics of entamoebas in wild NHP and for determining an accurate prevalence ofEntamoebaspecies.

Although wild chimpanzees and other African great apes live in regions endemic for African sleeping sickness, very little is known about their trypanosome infections, mainly due to major difficulties in obtaining their blood samples. In present work, we established a diagnostic ITS1-based PCR assay that allows detection of the DNA of all four Trypanosoma brucei subspecies (Trypanosoma bruceibrucei, Trypanosoma bruceirhodesiense, Trypanosoma bruceigambiense, and Trypanosoma bruceievansi) in feces of experimentally infected mice. Next, using this assay we revealed the presence of trypanosomes in the fecal samples of wild chimpanzees and this finding was further supported by results obtained using a set of primate tissue samples. Phylogenetic analysis of the ITS1 region showed that the majority of obtained sequences fell into the robust T. brucei group, providing strong evidence that these infections were caused by T. b. rhodesiense and/or T. b. gambiense. The optimized technique of trypanosome detection in feces will improve our knowledge about the epidemiology of trypanosomes in primates and possibly also other endangered mammals, from which blood and tissue samples cannot be obtained. Finally, we demonstrated that the mandrill serum was able to efficiently lyse T. b. brucei and T. b. rhodesiense, and to some extent T. b. gambiense, while the chimpanzee serum failed to lyse any of these subspecies., Graphical abstract, Highlights • ITS1-based PCR allows the detection of the Trypanosoma brucei in feces of non-human primates. • Wild chimpanzees are frequently infected with the T. brucei subspecies. • Mandrill serum efficiently lyses also Trypanosoma bruceigambiense.

Charismatic great apes have been used widely and effectively as flagship species in conservation campaigns for decades. These iconic representatives of their ecosystems could also play a role as reservoirs of several zoonotic diseases. Recently it was demonstrated that African great apes can host Leishmania parasites (Kinetoplastea: Trypanosomatidae). Given that this finding raised a strong negative reaction from leishmania experts and the subsequent discussion did not lead to a clear resolution, we decided to analyze wild gorilla (Gorilla gorilla gorilla) and chimpanzee (Pan troglodytes troglodytes) fecal samples collected from the same area in Cameroon as in the original study. Fecal samples, used to circumvent the difficulties and ethics involved in obtaining blood samples from endangered wild apes, were screened by three different PCR assays for detection of Leishmania DNA. We did not detect any leishmania parasites in analyzed feces; however, sequencing of SSU rRNA revealed an unexpected diversity of free-living bodonids (Kinetoplastea: Bodonidae) and parasitic trypanosomatids (Kinetoplastea: Trypanosomatidae) other than Leishmania. A single detected Phytomonas species, found in chimpanzee feces, most likely originated from animal plant food. On the other hand, the presence of four free-living bodonid species and four parasitic insect monoxenous trypanosomatid, including two possible new species of the genus Herpetomonas, could be explained as ex post contamination of feces either from the environment or from flies (Diptera: Brachycera). Keywords: Leishmania, Herpetomonas, Trypanosomatids, Detection, Gorilla, Chimpanzee, Feces, PCR

Objectives Environmental and ecological factors, such as geographic range, anthropogenic pressure, group identity, and feeding behavior are known to influence the gastrointestinal microbiomes of great apes. However, the influence of individual host traits such as age and sex, given specific dietary and social constraints, has been less studied. The objective of this investigation was to determine the associations between an individual's age and sex on the diversity and composition of the gut microbiome in wild western lowland gorillas. Materials and methods Publicly available 16S rRNA data generated from fecal samples of different groups of Gorilla gorilla gorilla in the Central African Republic were downloaded and bioinformatically processed. The groups analyzed included habituated, partially habituated and unhabituated gorillas, sampled during low fruit (dry, n = 28) and high fruit (wet, n = 82) seasons. Microbial community analyses (alpha and beta diversity and analyses of discriminant taxa), in tandem with network-wide approaches, were used to (a) mine for specific age and sex based differences in gut bacterial community composition and to (b) asses for gut community modularity and bacterial taxa with potential functional roles, in the context of seasonal food variation, and social group affiliation. Results Both age and sex significantly influenced gut microbiome diversity and composition in wild western lowland gorillas. However, the largest differences were observed between infants and adults in habituated groups and between adults and immature gorillas within all groups, and across dry and wet seasons. Specifically, although adults always showed greater bacterial richness than infants and immature gorillas, network-wide analyses showed higher microbial community complexity and modularity in the infant gorilla gut. Sex-based microbiome differences were not evident among adults, being only detected among immature gorillas. Conclusions The results presented point to a dynamic gut microbiome in Gorilla spp., associated with ontogeny and individual development. Of note, the gut microbiomes of breastfeeding infants seemed to reflect early exposure to complex, herbaceous vegetation. Whether increased compositional complexity of the infant gorilla gut microbiome is an adaptive response to an energy-limited diet and an underdeveloped gut needs to be further tested. Overall, age and sex based gut microbiome differences, as shown here, maybe mainly attributed to access to specific feeding sources, and social interactions between individuals within groups.

Strongylid nematodes in large terrestrial herbivores such as great apes, equids, elephants, and humans tend to occur in complex communities. However, identification of all species within strongylid communities using traditional methods based on coproscopy or single nematode amplification and sequencing is virtually impossible. High-throughput sequencing (HTS) technologies provide opportunities to generate large amounts of sequence data and enable analyses of samples containing a mixture of DNA from multiple species/genotypes. We designed and tested an HTS approach for strain-level identification of gastrointestinal strongylids using ITS-2 metabarcoding at the MiSeq Illumina platform in samples from two free-ranging non-human primate species inhabiting the same environment, but differing significantly in their host traits and ecology. Although we observed overlapping of particular haplotypes, overall the studied primate species differed in their strongylid nematode community composition. Using HTS, we revealed hidden diversity in the strongylid nematode communities in non-human primates, more than one haplotype was found in more than 90% of samples and coinfections of more than one putative species occurred in 80% of samples. In conclusion, the HTS approach on strongylid nematodes, preferably using fecal samples, represents a time and cost-efficient way of studying strongylid communities and provides a resolution superior to traditional approaches.

Made available in DSpace on 2018-12-11T16:52:00Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-04-01 Four species of Mammomonogamus are known from large African herbivores. A recent study demonstrated that a single Mammomonogamus species was shared by both western lowland gorillas (Gorilla gorilla gorilla) and African forest elephants (Loxodonta cyclotis) in Central African Republic, suggesting lower species diversity than previously described in literature. We examined more than 500 fecal samples collected from sympatric African forest elephants, western lowland gorillas, and African forest buffaloes (Syncerus caffer nanus) at four study sites across Central Africa and examined them by coproscopic methods to detect Mammomonogamus eggs, which were found at three of the study sites. Subsequently, sequences of 18S rDNA, 28S rDNA, and cox1 amplified from individual eggs were analyzed. Phylogenetic analyses of both nuclear and mitochondrial DNA revealed two clades: one formed by sequences originating from Gabonese buffaloes and the other comprising gorillas and elephants. The gorilla–elephant clade was further differentiated depending on the locality. We show the existence of at least two distinct species of Mammomonogamus, M. loxodontis in elephants and gorillas and M. nasicola in buffaloes. The available information on Mammomonogamus in African herbivores is reviewed. Department of Pathology and Parasitology Faculty of Veterinary Medicine University of Veterinary and Pharmaceutical Sciences Brno, Palackého tř. 1946/1 Central European Institute for Technology (CEITEC) University of Veterinary and Pharmaceutical Sciences, Palackého tř. 1946/1 Department of Virology Veterinary Research Institute, Hudcova 296/70 Institute of Vertebrate Biology Czech Academy of Sciences, Květná 8 Evolutionary Ecology and Conservation Genomics University of Ulm, Albert Einstein Allee 11 Institute of Parasitology Biology Centre of the Czech Academy of Sciences, Branišovská 31 Liberec Zoo, Lidové sady 425/1 WWF Dzanga-Sangha Protected Areas Projet Grands Singes Centre for Research and Conservation Royal Zoological Society of Antwerp, Koningin Astridplein 20–26 WWF Kudu-Zumbo Programme Universidade Estadual Paulista—UNESP Faculdade de Ciências Agrárias e Veterinárias Câmpus de Jaboticabal, Via de Acesso Prof. Paulo Donato Castelane, S/N Veterinary Medicine and Animal Science School UNESP—São Paulo State University, Rua Prof. Doutor Walter Mauricio Correa, S/N Universidade Estadual Paulista—UNESP Faculdade de Ciências Agrárias e Veterinárias Câmpus de Jaboticabal, Via de Acesso Prof. Paulo Donato Castelane, S/N Veterinary Medicine and Animal Science School UNESP—São Paulo State University, Rua Prof. Doutor Walter Mauricio Correa, S/N

Affiliation of Klara J. Petrželkova was incorrectly assigned as 2, 9, 10 in the original version of this article when in fact it should have been 3, 9, 10. Correct affiliations are presented here.

Relationships between gastrointestinal parasites (GIPs) and the gastrointestinal microbiome (GIM) are widely discussed topics across mammalian species due to their possible impact on the host's health. GIPs may change the environment determining alterations in GIM composition. We evaluated the associations between GIP infections and fecal microbiome composition in two habituated and two unhabituated groups of wild western lowland gorillas (Gorilla g. gorilla) from Dzanga Sangha Protected Areas, Central African Republic. We examined 43 fecal samples for GIPs and quantified strongylid nematodes. We characterized fecal microbiome composition through 454 pyrosequencing of the V1-V3 region of the bacterial 16S rRNA gene. Entamoeba spp. infections were associated with significant differences in abundances of bacterial taxa that likely play important roles in nutrition and metabolism for the host, besides being characteristic members of the gorilla gut microbiome. We did not observe any relationships between relative abundances of several bacterial taxa and strongylid egg counts. Based on our findings, we suggest that there is a significant relationship between fecal microbiome and Entamoeba infection in wild gorillas. This study contributes to the overall knowledge about factors involved in modulating GIM communities in great apes.

Exposure to stressors can negatively impact the mammalian gastrointestinal microbiome (GIM). Here, we used 454 pyrosequencing of 16S rRNA bacterial gene amplicons to evaluate the impact of physiological stress, as evidenced by faecal glucocorticoid metabolites (FGCM; ng/g), on the GIM composition of free-ranging western lowland gorillas (Gorilla gorilla gorilla). Although we found no relationship between GIM alpha diversity (H) and FGCM levels, we observed a significant relationship between the relative abundances of particular bacterial taxa and FGCM levels. Specifically, members of the family Anaerolineaceae (ρ=0.4, FDR q=0.01), genus Clostridium cluster XIVb (ρ=0.35, FDR q=0.02) and genus Oscillibacter (ρ=0.35, FDR q=0.02) were positively correlated with FGCM levels. Thus, while exposure to stressors appears to be associated with minor changes in the gorilla GIM, the consequences of these changes are unknown. Our results may have implications for conservation biology as well as for our overall understanding of factors influencing the non-human primate GIM.

Tsetse and tabanid flies transmit several Trypanosoma species, some of which are human and livestock pathogens of major medical and socioeconomic impact in Africa. Recent advances in molecular techniques and phylogenetic analyses have revealed a growing diversity of previously unidentified tsetse-transmitted trypanosomes potentially pathogenic to livestock and/or other domestic animals as well as wildlife, including African great apes. To map the distribution, prevalence and co-occurrence of known and novel trypanosome species, we analyzed tsetse and tabanid flies collected in the primary forested part of the Dzanga-Sangha Protected Areas, Central African Republic, which hosts a broad spectrum of wildlife including primates and is virtually devoid of domestic animals. Altogether, 564 tsetse flies and 81 tabanid flies were individually screened for the presence of trypanosomes using 18S rRNA-specific nested PCR. Herein, we demonstrate that wildlife animals are parasitized by a surprisingly wide range of trypanosome species that in some cases may circulate via these insect vectors. While one-third of the examined tsetse flies harbored trypanosomes either from the Trypanosoma theileri, Trypanosoma congolense or Trypanosoma simiae complex, or one of the three new members of the genus Trypanosoma (strains 'Bai', 'Ngbanda' and 'Didon'), more than half of the tabanid flies exclusively carried T. theileri. To establish the putative vertebrate hosts of the novel trypanosome species, we further analyzed the provenance of blood meals of tsetse flies. DNA individually isolated from 1033 specimens of Glossina spp. and subjected to high-throughput library-based screening proved that most of the examined tsetse flies engorged on wild ruminants (buffalo, sitatunga, bongo), humans and suids. Moreover, they also fed (albeit more rarely) on other vertebrates, thus providing indirect but convincing evidence that trypanosomes can be transmitted via these vectors among a wide range of warm- and cold-blooded hosts.

Rural areas of Romania, particularly the localities covering Danube Delta, are still not sufficiently explored in terms of epidemiological aspects, despite the large density of domestic animals living in close contact with people and natural environment of the Danube Delta Biosphere Reserve. Between 2010 and 2012, a survey on equine piroplasmids species was performed in this area, using a total of 178 horses, 15 donkeys and 177 dogs from 18 localities inside and outside the Danube Delta. None of the sampled hosts showed any clinical symptoms typical for equine piroplasmoses. A 25.4% overall prevalence for both equine species of piroplasmids was detected by PCR. Detection by "catch-all" primers followed by multiplex PCR revealed 20.3% prevalence of T. equi, beside 2.2% of B. caballi and 3.0% of mixed infections for all examined animals. Based on sequencing of 67 PCR amplicons of 18S rRNA gene of T. equi, four genotypes (A, C, D and E) were detected, whereas four B. caballi sequences clustered within a single genotype (A). A single dog infected with T. equi was found and confirmed by sequencing, clustering within genotype D. Our study further proved limited host specificity of piroplasmid. Higher prevalence was revealed inside the Danube delta, presumably caused by different environmental conditions, different tick densities and lower availability of veterinary services.

Infectious diseases including those caused by parasites can be a major threat to the conservation of endangered species. There is thus a great need for studies describing parasite infections of these species in the wild. Here we present data on parasite diversity in an agile mangabey (Cercocebus agilis) group in Bai Hokou, Dzanga-Sangha Protected Areas (DSPA), Central African Republic. We coproscopically analyzed 140 mangabey fecal samples by concentration techniques (flotation and sedimentation). Agile mangabeys hosted a broad diversity of protistan parasites/commensals, namely amoebas (Entamoeba spp., Iodamoeba buetschlli), a Buxtonella-like ciliate and several parasitic helminths: strongylid and spirurid nematodes, Primasubulura sp., Enterobius sp., and Trichuris sp. Importantly, some of the detected parasite taxa might be of potential zoonotic importance, such as Entamoeba spp. and the helminths Enterobius sp., Trichuris sp., and strongylid nematodes. Detailed morphological examination of ciliate cysts found in mangabeys and comparison with cysts of Balantioides coli from domestic pigs showed no distinguishing structures, although significant differences in cyst size were recorded. Scanning or transmission electron microscopy combined with molecular taxonomy methods are needed to properly identify these ciliates. Further studies using molecular epidemiology are warranted to better understand cross-species transmission and the zoonotic potential of parasites in sympatric non-human primates and humans cohabiting DSPA.

Increased anthropogenic activity can result in parasite exchanges and/or general changes in parasite communities, imposing a health risk to great apes. We studied protist and helminth parasites of wild western lowland gorilla groups in different levels of habituation, alongside humans inhabiting Dzanga-Sangha Protected Areas in the Central African Republic. Faeces were collected yearly during November and December from 2007 to 2010 and monthly from November 2010 to October 2011. Protist and helminth infections were compared among gorilla groups habituated, under habituation and unhabituated, and the effect of host traits and seasonality was evaluated. Zoonotic potential of parasites found in humans was assessed. No significant differences in clinically important parasites among the groups in different stages of habituation were found, except for Entamoeba spp. However, humans were infected with four taxa which may overlap with taxa found in gorillas. Females were less infected with spirurids, and adults had higher intensities of infection of Mammomonogamus sp. We found seasonal differences in the prevalence of several parasite taxa, but most importantly, the intensity of infection of unidentified strongylids was higher in the dry season. This study highlights that habituation may not necessarily pose a greater risk of protist and helminth infections in gorilla groups.

Background Although a high genetic diversity of Plasmodium spp. circulating in great apes has been revealed recently due to non-invasive methods enabling detection in faecal samples, little is known about the actual mechanisms underlying the presence of Plasmodium DNA in faeces. Great apes are commonly infected by strongylid nematodes, including hookworms, which cause intestinal bleeding. The impact of strongylid infections on the detection of Plasmodium DNA in faeces was assessed in wild, western, lowland gorillas from Dzanga Sangha Protected Areas, Central African Republic and eastern chimpanzees from Kalinzu Forest Reserve, Uganda. Methods Fifty-one faecal samples from 22 habituated gorillas and 74 samples from 15 habituated chimpanzees were analysed using Cytochrome-b PCR assay and coprological methods. Results Overall, 26.4% of the analysed samples were positive for both Plasmodium spp. and strongylids. However, the results showed no significant impact of intensity of infections of strongylids on detection of Plasmodium DNA in gorilla and chimpanzee faeces. Conclusion Bleeding caused by strongylid nematode Necator spp. cannot explain the presence of Plasmodium DNA in ape faeces.

SUMMARYSyngamid strongylids of the genusMammomonogamusundoubtedly belong among the least known nematodes with apparent zoonotic potential and the real diversity of the genus remains hard to evaluate without extensive molecular data. Eggs ofMammomonogamussp. are frequently found in feces of African forest elephants (Loxodonta cyclotis) and western lowland gorillas (Gorilla gorilla gorilla) in Dzanga-Sangha Protected Areas. Using sedimentation-based coproscopic techniques, we found the eggs ofMammomonogamusin 19·7% elephant and 54·1% gorilla fecal samples with 8–55 and 1–24 eggs per gram of fecal sediment for elephants and gorillas, respectively. We used a combination of light microscopy, scanning electron microscopy and analysis of cytochromecoxidase subunit I (cox1) and a partial sequence of 18S rDNA isolated from single eggs to test the hypothesis of possibleMammomonogamusconspecificity in gorillas and elephants. Whereas 18S rDNA sequences were identical in both gorillas and elephants, we distinguished seven different haplotypes within thecox1. Two haplotypes were found in both gorillas and elephants suggesting sharing ofMammomonogamus.Assignment of the parasite toM. loxodontisis proposed. Provided sequences represent the first genomic data onMammomonogamusspp.

We carried out a calibration of FLOTAC for ciliates Troglodytella abrassarti and Neobalantidium coli based on the selection of a most appropriate flotation solutions, and we also tested its accuracy (i.e., number of detected stages out of known added number of stages to fecal samples) and sensitivity for trophozoites of both ciliates in chimpanzee feces and N. coli cysts in pig feces, compared the detection threshold of FLOTAC with MIF-based sedimentation, and, subsequently, tested the losses of ciliate stages during sample preparation. Nine flotation solutions were evaluated, and ZnSO4 solution (specific gravity [s.g.] 1.2) showed to be the most suitable for trophozoite detection, while Sheather's solution (s.g. 1.33) was selected as most suitable for cysts. The FLOTAC sensitivity in detection of both stages varied: for trophozoites, we found all samples were positive when the intensity of infection 10 trophozoites per gram and higher, whereas for cysts the sensitivity was lower. The accuracy of FLOTAC negatively correlated with infection intensity, and the merthiolate-iodine-formaldehyde sedimentation-based quantification had a lower detection threshold. We demonstrated additional losses of stages of T. abrassarti and N. coli due to their retention in the sediment, which is probably a major reason for discrepancies in the numbers of countable ciliates between both methods. In conclusion, the FLOTAC should not be considered as a gold standard for quantification of intestinal ciliates in primates; instead, we recommend the modified MIF method.

Understanding variability in patterns of parasite infections requires studies of multiple populations inhabiting a variety of habitats. Gastrointestinal parasites of chimpanzees (Pan troglodytes) have been studied extensively at several forested sites, but the parasite fauna of chimpanzees living in dry, open habitats is less well known. We studied the parasites of savanna chimpanzees (Pan troglodytes schweinfurthii) living in the Issa Valley, Ugalla (Tanzania). We examined 119 fresh fecal samples using standard coproscopical methods. We detected protozoans including Blastocystis sp., Entamoeba coli, E. histolytica/dispar, Iodamoeba buetschlii, Troglodytella abrassarti, and Troglocorys cava, but only two types of spirurid nematodes among the helminths. The parasites of the Ugalla chimpanzees differ from those of forest chimpanzees in the absence of Strongyloides sp. and strongylid nematodes and a high prevalence of spirurids. Strongylids and Strongyloides sp. have thin-shelled eggs and larvae, which develop in the external environment; thus they may not be able to survive for prolonged periods in the extreme environment of Ugalla. The Ugalla chimpanzees also live at a lower population density and exhibit a larger home range than forest chimpanzees, factors that may lead to lower exposure to infective nematode larvae. Spirurid eggs, however, have thick shells and a life cycle dependent on intermediary hosts, making their survival and transmission in such extreme conditions more feasible. These differences between parasite fauna of closed and open forest chimpanzees contribute to our understanding of the ecology of infectious disease, and have the potential to contribute to conservation policies and practices.

One of the major factors threatening chimpanzees (Pan troglodytes verus) in Guinea-Bissau is habitat fragmentation. Such fragmentation may cause changes in symbiont dynamics resulting in increased susceptibility to infection, changes in host specificity and virulence. We monitored gastrointestinal symbiotic fauna of three chimpanzee subpopulations living within Cantanhez National Park (CNP) in Guinea Bissau in the areas with different levels of anthropogenic fragmentation. Using standard coproscopical methods (merthiolate-iodine formalin concentration and Sheather's flotation) we examined 102 fecal samples and identified at least 13 different symbiotic genera (Troglodytella abrassarti, Troglocorys cava, Blastocystis spp., Entamoeba spp., Iodamoeba butschlii, Giardia intestinalis, Chilomastix mesnili, Bertiella sp., Probstmayria gombensis, unidentified strongylids, Strongyloides stercoralis, Strongyloides fuelleborni, and Trichuris sp.). The symbiotic fauna of the CNP chimpanzees is comparable to that reported for other wild chimpanzee populations, although CNP chimpanzees have a higher prevalence of Trichuris sp. Symbiont richness was higher in chimpanzee subpopulations living in fragmented forests compared to the community inhabiting continuous forest area. We reported significantly higher prevalence of G. intestinalis in chimpanzees from fragmented areas, which could be attributed to increased contact with humans and livestock.

SUMMARYMicroscopic diagnosis of equine piroplasmoses, caused by Theileria equi and Babesia caballi, is hindered by low parasitaemia during the latent phase of the infections. However, this constraint can be overcome by the application of PCR followed by sequencing. Out of 288 animals examined, the piroplasmid DNA was detected in 78 (27·1%). Multiplex PCR indicated that T. equi (18·8%) was more prevalent than B. caballi (7·3%), while mixed infections were conspicuously absent. Sequences of 69 PCR amplicons obtained by the ‘catch-all’ PCR were in concordance with those amplified by the multiplex strategy. Computed minimal adequate model analyses for both equine piroplasmid species separately showed a significant effect of host species and age in the case of T. equi, while in the B. caballi infections only the correlation with host sex was significant. Phylogenetic analyses inferred the occurrence of three genotypes of T. equi and B. caballi. Moreover, a novel genotype C of B. caballi was identified. The dendrogram based on obtained sequences of T. equi revealed possible speciation events. The infections with T. equi and B. caballi are enzootic in all ecozones of Jordan and different genotypes circulate wherever dense horse population exists.

Background Habitat types can affect vector and pathogen distribution and transmission dynamics. The prevalence and genetic diversity of Plasmodium spp. in two eastern chimpanzee populations—Kalinzu Forest Reserve, Uganda and Issa Valley, Tanzania—inhabiting different habitat types was investigated. As a follow up study the effect of host sex and age on infections patterns in Kalinzu Forest Reserve chimpanzees was determined. Methods Molecular methods were employed to detect Plasmodium DNA from faecal samples collected from savanna-woodland (Issa Valley) and forest (Kalinzu Forest Reserve) chimpanzee populations. Results Based on a Cytochrome-b PCR assay, 32 out of 160 Kalinzu chimpanzee faecal samples were positive for Plasmodium DNA, whilst no positive sample was detected in 171 Issa Valley chimpanzee faecal samples. Sequence analysis revealed that previously known Laverania species (Plasmodium reichenowi, Plasmodium billbrayi and Plasmodium billcollinsi) are circulating in the Kalinzu chimpanzees. A significantly higher proportion of young individuals were tested positive for infections, and switching of Plasmodium spp. was reported in one individual. Amongst the positive individuals sampled more than once, the success of amplification of Plasmodium DNA from faeces varied over sampling time. Conclusion The study showed marked differences in the prevalence of malaria parasites among free ranging chimpanzee populations living in different habitats. In addition, a clear pattern of Plasmodium infections with respect to host age was found. The results presented in this study contribute to understanding the ecological aspects underlying the malaria infections in the wild. Nevertheless, integrative long-term studies on vector abundance, Plasmodium diversity during different seasons between sites would provide more insight on the occurrence, distribution and ecology of these pathogens. Electronic supplementary material The online version of this article (doi:10.1186/s12936-016-1476-2) contains supplementary material, which is available to authorized users.

Background: In general, studies on the diversity of strongylid nematodes in endangered host species are complicated as material obtained by non-invasive sampling methods has limited value for generic and species identification. While egg morphology barely allows assignment to family, the morphology of cultivated infective third stage larvae provides a better resolution at the generic level but cannot be used for exact species identification. Morphology-based taxonomic approaches greatly depend on the examination of adult worms that are usually not available. Methods: Hookworm parasites in two European researchers, who participated in gorilla research in the Central African Republic, were expelled after anthelmintic treatment to the faeces, collected and morphologically examined. A male worm discharged naturally from a wild bonobo (Pan paniscus) in Congo was also examined for comparison. Results: Two species of Necator were identified in researchers' faecal material: Necator americanus (Stiles, 1902) and N. gorillae Noda & Yamada, 1964; the latter species differed in having a smaller body, smaller buccal cavity and shorter spicules with spade-shaped membranes situated distally. Males of N. gorillae also possessed unusual cuticular thickenings on the dorsal side of the prebursal region of the body. These characters, shared with the male worm from the bonobo, correspond well to the description of N. gorillae described from gorillas in Congo. Conclusions: Based on the morphology of the hookworms recovered in this study and previous molecular analyses of larvae developed from both humans and western lowland gorillas (Gorilla gorilla gorilla) from this locality, we conclude that the researchers became infected with gorilla hookworms during their stay in the field. This is the first report of infection with a Necator species other than N. americanus in humans.

Freezing is considered to be the best method for long-term storage of bacterial DNA from feces; however this method cannot be usually applied for samples of wild primates collected in the challenging conditions of the tropical forest. In order to find an alternative conservation method of fecal samples from wild great apes, we compared freezing with other fixation methods. Fecal samples from 11 captive gorillas (Gorilla gorilla gorilla) from three Czech Zoos were stored using freezing, RNA Stabilization Reagent (RNAlater), and 96% ethanol. Subsequently, the samples were examined using culture-independent methods (PCR-DGGE, and Real-time PCR) to qualitatively and quantitatively assess fecal microbiota composition and to compare differences among the storage methods. Noticeably, freezing samples resulted in the highest recoveries of DNA. No significant differences in DNA recovery were found between freezing and using RNAlater; however, significantly lower DNA concentrations were recovered from samples stored in 96% ethanol. Using PCR-DGGE we found that either 96% ethanol, RNAlater or freezing were suitable for preserving bacterial DNA; however fingerprints obtained from RNAlater storage were more similar to those obtained from the frozen method; in comparison to the patterns resulting from storing samples in ethanol. Using qPCR, frozen samples yielded the highest values of bacterial counts, with the exception of Enterobacteriaceae, which showed the highest numbers using samples stored in ethanol. Sequences of amplicons obtained from PCR-DGGE belonged to the families Clostridiaceae, Lactobacillaceae, Staphylococcaceae, and Lachnospiraceae, phylum Firmicutes; however most amplicons showed sequence similarity to previously uncultured microorganisms. Bacteria belonging to the phylum Firmicutes were the most frequently identified species in the fecal bacterial communities of captive western gorillas. The study showed that RNAlater is an optimal storage method when freezing is not possible.

The entodiniomorphid ciliate Troglodytella abrassarti is a colonic mutualist of great apes. Its host specificity makes it a suitable model for studies of primate evolution. We explored molecular diversity of T. abrassarti with regard to large geographical distribution and taxonomic diversity of its most common host, the chimpanzee. We found a very low diversification of T. abrassarti in chimpanzees across Africa. Distribution of two types of T. abrassarti supports evolutionary separation of the Western chimpanzee, P. t. verus, from populations in Central and East Africa. Type I T. abrassarti is probably a derived form, which corresponds with the Central African origin of chimpanzees and a founder event leading to P. t. verus. Exclusivity of the respective types of T. abrassarti to Western and Central/Eastern chimpanzees corroborates the difference found between an introduced population of presumed Western chimpanzees on Rubondo Island and an autochthonous population in mainland Tanzania. The identity of T. abrassarti from Nigerian P. t. ellioti and Central African chimpanzees suggests their close evolutionary relationship. Although this contrasts with published mtDNA data, it corroborates current opinion on the exclusive position of P. t. verus within the chimpanzee phylogeny. The type of T. abrassarti occurring in Central and East African common chimpanzee was confirmed also in bonobos. This may point to the presence of an ancestral Type II found throughout the Lower Guinean rainforest dating back to the common Pan ancestor. Alternatively, the molecular uniformity of T. abrassarti may imply a historical overlap of the species' distribution ranges.

Our results suggest, however, that infection intensities of T. abrassarti in captive chimpanzees are not influenced primarily by the amount of fiber in the diet, but rather by the dietary starch concentration (HFD: 1%; LFD: 8%)

SUMMARYDespite the fact that the non-human primates are our closest relatives and represent a species-rich mammalian group, little is known about their intestinal protistan parasites/commensals. Particularly, the intestinal trichomonads represent a neglected part of the fauna of the primate digestive system. We have established 30 trichomonad strains isolated from feces of 11 primate species kept in 3 Czech zoos and performed an analysis of their SSU rDNA and ITS1-5·8S rDNA-ITS2. Our results showed that intestinal trichomonads are rather common among non-human primates. Molecular phylogenetic analysis showed that the strains are unexpectedly diversified, belonging to 8 or 9 distinct species. Interestingly, the vast majority of the strains from non-human primates belonged to the genus Tetratrichomonas while no member of this genus has been found in the human intestine so far. In addition, hominoid and non-hominoid primates differed in their intestinal trichomonads. Our results suggest that captive primates possibly may be infected by intestinal trichomonads of other vertebrates such as pigs, cattle, birds, tortoises and lizards.

The isolated ecosystem of Rubondo Island National Park, Tanzania is an interesting model site, inhabited by an assembly of primate species with various histories: two introduced primate species, Pan troglodytes (chimpanzee) and Colobus guereza (colobus), and a single indigenous species Chlorocebus aethiops pygerythrus (vervet monkey). Apart from important lessons for future introduction/re-introduction projects, Rubondo National Park offers a unique place to study the patterns of transmission of primate parasites and their host specificity. Blastocystis was detected using standard microscopy, together with PCR-based determination and the prevalence and subtype identification of Blastocystis was determined in each primate species. Subtype (ST) 1 was detected in all three Rubondo primate populations; ST2, ST3 and ST5 were found in colobus and vervet monkeys. All chimpanzee isolates of Blastocystis belonged exclusively to ST1, which formed a discrete group, suggesting that Rubondo chimpanzees are colonized by a single, host-specific Blastocystis strain that circulates among the members of the group. Phylogenetic analyzes indicated that transmission of Blastocystis did not occur between Rubondo primate populations. Observed host specificity of Blastocystis provides a new understanding of the transmission and distribution of Blastocystis among sympatric hosts under natural conditions.