Dipetalonema graciliformis (Freitas, 1964) from the red-handed tamarins (Saguinus midas, Linnaeus, 1758) in French Guiana.

Six Dipetalonema species have been reported from Neotropical monkeys, Dipetalonema gracile, Dipetalonema graciliformis and Dipetalonema caudispina being the dominant species found in French Guiana primates. Adult filarioids isolated from the abdominal cavity of tamarins (Saguinus midas) in French Guiana were morphologically and molecularly identified as D. graciliformis. Phylogenetic analysis based on DNA and amino acid sequences of the cox1 gene as well as the concatenated sequences of the cox1 and the 18S genes indicated that D. graciliformis belongs to the clade 4 (ONC4) of Onchocercidae. Blast analysis of the 18S rDNA revealed that D. graciliformis in the studied tamarins is conspecific with the filarioid circulating in howler monkeys (Alouatta macconnelli) in French Guiana, previously referred to as unidentified Onchocercidae species.

Chronic Plasmodium brasilianum infections in wild Peruvian tamarins.

There is an increased interest in potential zoonotic malarias. To date, Plasmodium malariae that infects humans remains indistinguishable from Plasmodium brasilianum, which is widespread among New World primates. Distributed throughout tropical Central and South America, the Callitrichidae are small arboreal primates in which detection of natural Plasmodium infection has been extremely rare. Most prior screening efforts have been limited to small samples, the use of low-probability detection methods, or both. Rarely have screening efforts implemented a longitudinal sampling design. Through an annual mark-recapture program of two sympatric callitrichids, the emperor (Saguinus imperator) and saddleback (Saguinus fuscicollis) tamarins, whole blood samples were screened for Plasmodium by microscopy and nested PCR of the cytochrome b gene across four consecutive years (2012-2015). Following the first field season, approximately 50% of the samples collected each subsequent year were from recaptured individuals. In particular, out of 245 samples from 129 individuals, 11 samples from 6 individuals were positive for Plasmodium, and all but one of these infections was found in S. imperator. Importantly, the cytochrome b sequences were 100% identical to former isolates of P. malariae from humans and P. brasilianum from Saimiri sp. Chronic infections were detected as evidenced by repeated infections (7) from two individuals across the 4-year study period. Furthermore, 4 of the 5 infected emperor tamarins were part of a single group spanning the entire study period. Overall, the low prevalence reported here is consistent with previous findings. This study identifies two new natural hosts for P. brasilianum and provides evidence in support of chronic infections in wildlife populations. Given that callitrichids are often found in mixed-species associations with other primates and can be resilient to human-disturbed environments, they could contribute to the maintenance of P. malariae populations if future work provides entomological and epidemiological evidence indicating human zoonotic infections.

The influence of human settlements on the parasite community in two species of Peruvian tamarin.

Although there is a growing recognition that the transfer of diseases between humans and non-human primates can be of great significance for conservation biology, there have been only a few studies focusing on parasites. In this study, saddleback (Saguinus fuscicollis) and moustached tamarin (Saguinus mystax) from the rainforest of the Peruvian lowlands were used as models to determine helminth parasite associations between canopy-dwelling primate species and a nearby human settlement. The human population showed high prevalences of infestation with a number of nematodes, including Ascaris lumbricoides (88.9%), Trichuris trichiura (37%) and hookworms (55.6%). However, the ova of these geohelminths were not detectable in tamarin faeces. Thus, no direct parasite transfer from humans to non-human primates could be documented. However, tamarin groups with more frequent contact to humans and their facilities had significantly higher prevalences and egg output of Prosthenorchis elegans, an important primate pathogen, than a forest group. In contrast, a cestode was significantly more common with more egg output in sylvatic than in human-associated groups. Human alteration of the habitat is likely to play a major role in determining the occurrence, prevalence and intensity of helminth infestation of wild non-human primates.

Trypanosoma cruzi in Brazilian Amazonia: Lineages TCI and TCIIa in wild primates, Rhodnius spp. and in humans with Chagas disease associated with oral transmission.

In this study, we provide phylogenetic and biogeographic evidence that the Trypanosoma cruzi lineages T. cruzi I (TCI) and T. cruzi IIa (TCIIa) circulate amongst non-human primates in Brazilian Amazonia, and are transmitted by Rhodnius species in overlapping arboreal transmission cycles, sporadically infecting humans. TCI presented higher prevalence rates, and no lineages other than TCI and TCIIa were found in this study in wild monkeys and Rhodnius from the Amazonian region. We characterised TCI and TCIIa from wild primates (16 TCI and five TCIIa), Rhodnius spp. (13 TCI and nine TCIIa), and humans with Chagas disease associated with oral transmission (14 TCI and five TCIIa) in Brazilian Amazonia. To our knowledge, TCIIa had not been associated with wild monkeys until now. Polymorphisms of ssrDNA, cytochrome b gene sequences and randomly amplified polymorphic DNA (RAPD) patterns clearly separated TCIIa from TCIIb-e and TCI lineages, and disclosed small intra-lineage polymorphisms amongst isolates from Amazonia. These data are important in understanding the complexity of the transmission cycles, genetic structure, and evolutionary history of T. cruzi populations circulating in Amazonia, and they contribute to both the unravelling of human infection routes and the pathological peculiarities of Chagas disease in this region.

Infection rates and genotypes of Trypanosoma rangeli and T. cruzi infecting free-ranging Saguinus bicolor (Callitrichidae), a critically endangered primate of the Amazon Rainforest.

Parasites of wild primates are important for conservation biology and human health due to their high potential to infect humans. In the Amazon region, non-human primates are commonly infected by Trypanosoma cruzi and T. rangeli, which are also infective to man and several mammals. This is the first survey of trypanosomiasis in a critically endangered species of tamarin, Saguinus bicolor (Callitrichidae), from the Brazilian Amazon Rainforest. Of the 96 free-ranging specimens of S. bicolor examined 45 (46.8%) yielded blood smears positive for trypanosomes. T. rangeli was detected in blood smears of 38 monkeys (39.6%) whereas T. cruzi was never detected. Seven animals (7.3%) presented trypanosomes of the subgenus Megatrypanum. Hemocultures detected 84 positive tamarins (87.5%). Seventy-two of 84 (85.7%) were morphologically diagnosed as T. rangeli and 3 (3.1%) as T. cruzi. Nine tamarins (9.4%) yielded mixed cultures of these two species, which after successive passages generated six cultures exclusively of T. cruzi and two of T. rangeli, with only one culture remaining mixed. Of the 72 cultures positive for T. rangeli, 62 remained as established cultures and were genotyped: 8 were assigned to phylogenetic lineage A (12.9%) and 54 to lineage B (87.1%). Ten established cultures of T. cruzi were genotyped as TCI lineage (100%). Transmission of both trypanosome species, their potential risk to this endangered species and the role of wild primates as reservoirs for trypanosomes infective to humans are discussed.

Prevalence and molecular phylogenetic characterization of Trypanosoma (Megatrypanum) minasense in the peripheral blood of small neotropical primates after a quarantine period.

Neotropical primates of the Cebidae and Callitrichidae, in their natural habitats, are frequently infected with a variety of trypanosomes including Trypanosoma cruzi, which causes a serious zoonosis, Chagas' disease. The state of trypanosome infection after a 30-day quarantine period was assessed in 85 squirrel monkeys (Saimiri sciureus) and 15 red-handed tamarins (Saguinus midas), that were wild-caught and exported to Japan as companion animals or laboratory animals, for biomedical research, respectively. In addition to many microfilariae of Mansonella (Tetrapetalonema) mariae at a prevalence of 25.9%, and Dipetalonema caudispina at a prevalence of 3.5%, a few trypomastigotes of Trypanosoma (Megatrypanum) minasense were detected in Giemsa-stained thin films of blood from 20 squirrel monkeys at a prevalence of 23.5%. Although few T. minasense trypomastigotes were found in Giemsa-stained blood films from tamarins, a buffy-coat examination detected trypanosomes in 12 red-handed tamarins (80.0%), and PCR amplification of a highly variable region of the small subunit ribosomal RNA genes (SSU rDNA) for Trypanosoma spp. detected the infection in 14 of the 15 tamarins (93.3%). Nucleotide sequences of the amplicons were identical for trypanosomes from tamarins and squirrel monkeys, indicating a high prevalence but low parasitemia of T. minasense in imported Neotropical nonhuman primates. Based on the SSU rDNA and 5.8S rDNA, the molecular phylogenetic characterization of T. minasense indicated that T. minasense is closely related to trypanosomes with Trypanosoma theileri-like morphology and is distinct from Trypanosoma (Tejeraia) rangeli, as well as from T. cruzi. Using some blood samples from these monkeys, amplification and subsequent sequencing of the glycosomal glyceraldehyde-3-phosphate dehydrogenase (gGAPDH) gene fragments detected 4 trypanosome genotypes, including 2 types of T. cruzi clade, 1 type of T. rangeli clade, and 1 T. rangeli-related type, but failed to indicate its phylogenetic position based on the gGAPDH gene. Furthermore, species ordinarily classified in the Megatrypanum by morphological criteria do not form a clade in any molecular phylogenetic trees based on rDNA or gGAPDH genes.

Disseminated lethal Encephalitozoon cuniculi (genotype III) infections in cotton-top tamarins (Oedipomidas oedipus)--a case report.

For the first time, Encephalitozoon (E.) cuniculi genotype III ('dog strain') was verified in two cotton-top tamarins (Oedipomidas oedipus) by light microscopy, immunohistochemistry, electron microscopy, PCR and sequencing. The animals had a disseminated lethal infection with this protist. In earlier reports, genotype III had been found only in domestic dogs, man, emperor tamarins (Saguinus imperator) and golden lion tamarins (Leontopithecus rosalia). This investigation establishes now that the 'dog strain' can occur in cotton-top tamarins too. This is further evidence for the zoonotic potential of E. cuniculi. Furthermore, free E. cuniculi spores were identified also in blood vessels of several tissues. These findings indicate that during a disseminated infection E. cuniculi spores can occur in peripheral blood, too. We propose that blood should also be included in the investigations for the detection of microsporidia, so that a possible disseminated course of an infection can be detected.

Overt fatal and chronic subclinical Encephalitozoon cuniculi microsporidiosis in a colony of captive emperor tamarins (Saguinus imperator).

The course of an infection with the microsporidian Encephalitozoon cuniculi in a colony of captive emperor tamarins (Saguinus imperator) is described. In two litters, the infection was associated with overt disease and death of all infants. Immunohistochemistry for E. cuniculi showed generalized infections, and histopathologic evaluation revealed systemic vasculitis and disseminated mixed inflammatory cell infiltration with and without necrosis in several organs. Serologically, some of the juvenile animals presented with high titres for Encephalitozoon, while the adults had low titres. The E. cuniculi "dog strain" was identified by molecular means for the first time in Europe. The origin of the infection appeared to be a pair of breeding adults that originated from the US. Our data suggest that the organism persisted over years in the colony, and that subclinically infected animals most likely were involved in perpetuating the infection. Efforts should be made to ascertain if this microorganism is present in other captive populations of this endangered monkey species and to prevent its further spreading.

Pterygodermatites nycticebi (Nematoda: Rictulariidae): accidental detection of encapsulated third-stage larvae in the tissue of a white-fronted marmoset.

Twin, white-fronted marmosets (Callithrix geoffroyi) born and raised in a zoo in Japan died at 7 mo of age. Several encapsulated nematode larvae were detected in the intestinal wall, as well as a few in the mesenteric lymph nodes of 1 of the twins. In the other marmoset, no encapsulated nematode larva was detected in the organs, but many adult Pterygodermatites nycticebi were found in the intestinal lumen. In the past 5 yr, 5 primates kept in the same zoo, i.e., 1 squirrel monkey (Saimiri sciureus), 2 Pygmy marmosets (Cebuella pygmaea), 1 Senegal galago (Galago senegalensis), and 1 cotton-top tamarin (Saguinus oedipus), died from heavy infestation with the same nematode. A few migrating larvae of the rictulariid were also identified histologically in the intestinal wall and liver of the cotton-top tamarin. Although no other primate currently held in the same zoo was infected with the rictulariid, German cockroaches (Blattella germanica) collected with traps near marmoset cages had encapsulated P. nycticebi larvae, indicating latent perpetuation of the life cycle of this rictulariid species in the zoo premises. Our results indicated that encapsulation or migration of third-stage larvae of P. nycticebi might occur accidentally in the organs of callithrichid primates.

Toxoplasmosis in golden-headed lion tamarins (Leontopithecus chrysomelas) and emperor marmosets (Saguinus imperator) in captivity.

From 1991 to 1995, eight New World nonhuman primates of the family Callitrichidae belonging to the collection of Fundacão Parque Zoologico de São Paulo died of toxoplasmosis. Of the eight affected nonhuman primates, four were Leontopithecus chrysomelas (one male, three females) and four were Saguinus imperator (two males, two females). The most commonly affected organs were the lungs, liver, and lymph nodes, with hemorrhagic and necrotic lesions. Histopathologic examination revealed protozoa that were morphologically consistent with Toxoplasma gondii. Immunohistochemical assays were strongly positive for T. gondii.

Demodicosis in red-handed tamarins (Saguinus midas).

Two nonrelated but paired red-handed tamarins (Saguinus midas) presented with diffuse, multifocal, raised, nonpruritic, hyperkeratotic lesions on the appendages and face. Skin biopsies identified acarids and skin scrapings confirmed demodex-like mites. The animals were treated with ivermectin, at the endoparasite dose, which initially resulted in resolution of clinical signs; however, signs recurred after numerous treatments. After four treatments with amitraz dips, demodicosis lesions resolved.

The complexity of the sylvatic cycle of Trypanosoma cruzi in Rio de Janeiro state (Brazil) revealed by the non-transcribed spacer of the mini-exon gene.

American trypanosamiasis occurs in nature as a sylvatic cycle, where Trypanosoma cruzi interacts with wild triatomines and mammalian reservoirs, such as marsupials, rodents, armadillos and other animals. Due to difficulties in trying to isolate T. cruzi stocks from the sylvatic cycle, very few studies have been performed in order to understand the parasite infection in natural environments. Traditionally T. cruzi has been considered to be composed of a highly heterogeneous population of parasites. In contrast, the mini-exon and the 24S alpha rRNA gene loci have shown that T. cruzi stocks can be clustered in 2 major phylogenetic groups: lineage 1 and lineage 2. In this report, 68 recently isolated T. cruzi samples from the sylvatic cycle belonging to different geographical areas in Rio de Janeiro, Brazil, have been typed based on a variable spot in the non-transcribed spacer of the mini-exon gene. Eight isolates were from triatomines, 26 stocks were from golden-lion tamarins, 31 from opossums, 2 from rodents and 1 from a three-toed sloth. Thirty (44%-30/68) isolates were typed as lineage 1, while 36 (53%-36/68) isolates were typed as lineage 2. Two opossums presented mixed infection. Therefore, 3% (2/68) of the isolates were typed as lineage 1 + lineage 2. Using these geographical regions as models of sylvatic environments, it was observed that 96% of the Didelphis marsupialis were infected by lineage 2 isolates, while all 26 golden-lion tamarins were infected by lineage 1. The results show preferential association of the 2 lineages of T. cruzi with different hosts, composing the complexity of the sylvatic cycle.

Cerebrospinal angiostrongyliasis in five captive tamarins (Sanguinus spp).

Four cotton-top tamarins (Sanguinus oedipus oedipus) and one emperor tamarin (S imperator subgrisescens) housed in a zoo became depressed, anorexic, paraparetic and eventually paralysed. The animals died within 5 days to 18 months of the appearance of clinical signs. Histological examination showed nonsuppurative and eosinophilic meningoencephalitis, and metastrongyle nematode larvae were found within subarachnoid spaces of all animals and within the spinal cord of one. Intact larvae with features consistent with Angiostrongylus cantonensis were recovered from the brain of one animal. This parasite is the classical cause of eosinophilic meningoencephalitis in many parts of the world and the diagnosis can be strongly suspected on clinical grounds. In endemic areas like south-east Queensland, protection of captive animals against infection with A cantonensis is a difficult balance between providing a stimulating, natural setting and eliminating potentially infectious definitive, intermediate and paratenic hosts. This is the first report of cerebrospinal angiostrongyliasis in tamarins and nonhuman primates in Australia.

Experimental Brugia pahangi and B. malayi infections of callitrichid primates.

The callitrichid primates, Callithrix jacchus jacchus (the marmoset) and Saguinus labiatus (the tamarin) were inoculated with infective larvae of Brugia malayi and B. pahangi. Microfilaraemia at low levels developed in 3 out of 4 C.j. jacchus infected with B. malayi and living or dead adult worms found in all 4. Only one of 4 C.j. jacchus became microfilaraemic (mf + ve) when given B. pahangi and adults were found in two. Of 4 S. labiatus given B. pahangi one became very lightly mf + ve and adults were found in 3. It is concluded that these animals are not suitable hosts for chemotherapeutic experiments.

Experimental Schistosoma mansoni infection in a small New World monkey, the saddle-back tamarin (Saguinus fuscicollis).

Twelve young adult captivity-born tamarin monkeys (Saguinus fuscicollis) were each exposed to 150 cercariae of Schistosoma mansoni (KEB strain): 6 by the percutaneous (pc) and 6 by the subcutaneous (sc) route. The prepatent period, as determined by eggs in the feces, was 34-39 days for both groups. Weekly quantitative fecal examinations revealed that although both groups actively passed eggs for as long as the duration of the experiment (18 months), the sc group excreted a significantly higher number of eggs/gram/day in the feces than did the pc group. Eggs recovered from the feces of both groups were viable: they hatched and the miracidia invaded snails which subsequently yielded infective cercariae. A significantly greater number of worms was recovered from tamarins infected by the sc route as compared to those with pc infections, and large numbers of eggs were found in affected organs of the sc group. Chronically infected tamarins with high tissue egg loads developed focal granulomatous nodules along the serosal walls of the small intestines, which in some cases, apparently had budded off to lie free in the abdominal cavity. Hepatic granulomas from animals with acute infections were significantly larger than those from chronically infected monkeys. Our results strongly suggest the presence of a skin barrier to infection in the tamarin monkey, and that if this barrier is bypassed, the tamarin can serve as a permissive host for S. mansoni. The small body size of this monkey (less than 400 g) is an obvious reason for the establishment of the tamarin monkey as a laboratory nonhuman primate model for human schistosomiasis mansoni.

Trypanosoma cruzi and Trypanosoma rangeli in Saimiri sciureus from Bolivia and Saguinus mistax from Brazil.

Examination of blood films for trypanosomes in primates housed at the Tulane University Delta Regional Primate Research Center showed that 47% (32/68) of the Bolivian Saimiri sciureus and 51% (68/135) of Brazilian Saguinus mistax harbored one or more types: Trypanosoma (Schizotrypanum) cruzi in 6% to 7% and Trypanosoma (Herpetosoma) spp. or Trypanosoma (Megatrypanum) spp. in 39% and 45%, respectively. Trypanosomes were isolated from a sample of the infected monkeys and morphobiological studies were carried out. Both T. cruzi and Trypanosoma rangeli were demonstrated. The transmission of T. rangeli by Rhodnius prolixus, Rhodnius neglectus, and Triatoma infestans was assessed: only R. prolixus transmitted the infection by bite. T. rangeli has not been found previously in Bolivia, nor has any trypanosome been reported previously in Saguinus mistax.

Anion-exchange separation for neotropical trypanosomes: a preliminary trial and a description of Trypanosoma devei from the tamarin Saguinus midas niger.

Anion-exchange separation trials using DEAE-cellulose columns were performed with blood from two single species of marsupial and edentate, three species of rodent and single species of carnivore, primate, cayman and lizard. Trypanosoma cruzi was isolated from Didelphis marsupialis, Dasypus novemcinctus and Coendou sp. T. (Megatrypanum) devei was isolated from the tamarin Saguinus midas niger and the mensural characters of the organism were redescribed. Anion-exchange separation was considered to be a valuable procedure for the taxonomist searching for new or little-known trypanosomes.

Spontaneous occurrence of Angiostrongylus costaricensis in marmosets (Saguinus mystax).

Two marmosets imported from Iquitos, Peru, were found to be infected with Angiostrongylus costaricensis. Both animals had large solitary granulomas involving the wall and adjacent mesentery of the small intestine. Histopathologic examination showed the adult nematodes in the lumina of the mesenteric arteries that coursed through these granulomas. The inflammatory reaction was associated with numerous degenerating eggs and larvae. This is the first report of this parasite in nonhuman primates and extends its geographic range to Peru. In addition, in one animal, Dipetalonema sp were seen free in the abdominal cavity, and pleroceroid larvae (spargana) were in the loose connective tissue of the left axilla. This animal also had microgranulomas associated with eggs and larvae of Angiostrongylus in the kidney, liver, lung, and heart.