Frequency of enteroparasites in Cebidae and Callitrichidae primates at the Zoológico de Cali, Colombia: zoonotic implications / [Frecuencia de enteroparásitos en primates Cebidae y Callitrichidae del Zoológico de Cali, Colombia: implicaciones zoonóticas].

Introduction: Enteroparasites can cause problems in animals kept under human care in zoos and shelters. Wild animals have low parasitic loads but when sheltered in closed places they can be higher and lead to clinical manifestations, which increases the cost of medical treatments and care. On the other hand, some enteroparasites can represent a potential risk of zoonotic infection for their animal keepers, visitors, and other zoo animals. In addition, they could affect recovery programs for endangered species. Objectives: To establish the presence and prevalence of potentially zoonotic enteroparasites in primates of the Cebidae and Callitrichidae families at the Zoológico de Cali from September to November, 2017. Materials and methods: We conducted a prospective cross-sectional study. Serial samples from 50 individuals belonging to seven species and two primate families were analyzed by ova and parasite test, flotation, and Kinyoun stain between September and November, 2017. Results: In order of frequency, the parasite genera found in the seven primate species evaluated were Blastocystis spp., Trichomonas spp., Giardia spp., Entamoeba spp., Strongyloides spp., Cyclospora sp., and Trichuris sp. Conclusions: At least six of the parasite genera found have potential zoonotic implications. It is necessary to establish what are the infection sources at the Zoológico de Cali and implement management protocols to reduce the risk of transmission to both humans and other animals in the collection. Additionally, we offer relevant information on the zoonotic potential of each of the enteroparasites found.

Introducción. Los enteroparásitos pueden generar problemas en animales bajo cuidado humano en zoológicos y centros de acogida. Los animales silvestres presentan bajas cargas parasitarias, pero estas pueden ser mayores y llevar a manifestaciones clínicas cuando se trata de animales resguardados en recintos, lo que aumenta los gastos en tratamientos y cuidados médicos. Por otro lado, algunos enteroparásitos pueden causar infecciones zoonóticas en los cuidadores, los visitantes y otros animales del zoológico, así como afectar los programas de recuperación de especies amenazadas de extinción. Objetivos. Determinar la presencia y prevalencia de enteroparásitos con potencial de transmisión zoonótica en primates de las familias Cebidae y Callitrichidae del Zoológico de Cali, entre septiembre y noviembre de 2017. Materiales y métodos. Se hizo un estudio transversal prospectivo, para lo cual se recolectaron muestras seriadas de 50 individuos pertenecientes a siete especies de dos familias de primates y se analizaron mediante examen coprológico, flotación y coloración Kinyoun, entre septiembre y noviembre de 2017. Resultados. Según su prevalencia, los géneros de parásitos hallados en las siete especies de primates evaluadas, fueron Blastocystis spp., Trichomonas spp., Giardia spp., Entamoeba spp., Strongyloides spp., Cyclospora sp. y Trichuris sp. Conclusiones. Por lo menos, seis de los géneros de parásitos identificados tienen implicaciones zoonóticas, lo cual hace necesario establecer las posibles vías de infección de los primates del Zoológico de Cali e implementar protocolos de manejo que reduzcan el riesgo de transmisión a los humanos y a otros animales de la colección. Además, se presenta la información relevante sobre el potencial zoonótico de los enteroparásitos hallados.

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.

Aotus lemurinus griseimembra monkeys: a suitable model for Plasmodium vivax sporozoite infection.

This study describes a successful Plasmodium vivax sporozoite infection in Aotus lemurinus griseimembra. Twenty-eight naive or previously infected monkeys, either splenectomized or spleen intact, were inoculated intravenously or subcutaneously with Plasmodium vivax sporozoites of the Salvador I strain or with two wild isolates (VCC-4 and VCC-5; Vivax-Cali-Colombia). The monkeys were successfully infected regardless of the parasite strain, spleen presence, or inoculation route and showed prepatent periods that ranged from 16 to 89 days. Only one monkey inoculated intravenously failed to develop parasitemia. Since immune protection against malaria pre-erythrocytic forms is mediated by both helper and cytolytic T cells that may home in the spleen and P. vivax cultures are not yet available; the use of spleen-intact A. lemurinus griseimembra, susceptible to both adapted and non-adapted strains of P. vivax sporozoites, is a valuable model for evaluation of pre-erythrocytic vaccine candidates.

Entamoeba histolytica infections in captive primates.

A group based survey on the presence of Entamoeba histolytica and Entamoeba dispar using real-time PCR among 20 species of captive non-human primates was performed after diagnosis of E. histolytica dysentery in a spider monkey ( Ateles belzebuth hybridus). E. histolytica DNA was detected in three species of New World primates and in three species of Old World primates. In five of six E. histolytica isolates, it was possible to amplify the SREHP gene. They all revealed the same pattern after AluI digestion, indicating a common source of infection. E. dispar DNA was detected in two species of New World monkeys and three species of Old World monkeys. The results demonstrate that E. histolytica is capable of causing symptomatic and non-symptomatic infections in Old World and New World non-human primates. To our knowledge, this is the first report of E. histolytica sensu stricto in non-human primates after the redescription separating it from E. dispar in 1993.

Transmission dynamics of Cryptosporidium in primates and herbivores at the Barcelona zoo: a long-term study.

Factors influencing the transmission of Cryptosporidium in primates and herbivores housed at the Barcelona zoo have been analyzed. The relationship between continuous and discontinuous oocyst shedding, both animal housing conditions and abiotic factors (seasonality, humidity, temperature) was examined to explain the epizootiology of the protozoan. Thirty six fecal samples from each of 11 primates (Pongidae, Cebidae, Cercopithecidae and Lemuridae) and 22 herbivores (Elephantidae, Camelidae, Cervidae, Giraffidae and Bovidae) were examined over the period of 1 year. The parasite transmission was based on the chronic infection status of some animals serving as a source of successive reinfection for other animals. The environmental temperature and humidity (seasonality), the physical features of the facilities, the vicinity of the animals and the physiological status induced by captivity contributed to transmission. The long-term character of this study was essential for obtaining these results and interpreting the complex relationships.

Adaptation of a strain of Plasmodium vivax from India to New World monkeys, chimpanzees, and anopheline mosquitoes.

A strain of Plasmodium vivax from India was adapted to develop in splenectomized Saimiri boliviensis, Aotus lemurinus griseimembra, A vociferans, A. nancymai, A. azarae boliviensis, hybrid Aotus monkeys, and splenectomized chimpanzees. Infections were induced via the inoculation of sporozoites dissected from the salivary glands of Anopheles stephensi and An. dirus mosquitoes to 12 Aotus and 8 Saimiri monkeys; transmission via the bites of infected An. stephensi was made to 1 Aotus monkey and 1 chimpanzee. The intravenous passage of infected erythrocytes was made to 9 Aotus monkeys and 4 chimpanzees. Gametocytes in 13 Aotus monkeys and 4 chimpanzees were infectious to mosquitoes. Infection rates were markedly higher in mosquitoes fed on chimpanzees. PCR studies on 10 monkeys injected with sporozoites revealed the presence of parasites before their detection by microscopic examination. The India VII strain of P. vivax develops in Aotus and Saimiri monkeys and chimpanzees following the injection of parasitized erythrocytes, or sporozoites, or both. The transmission rate via sporozoites to New World monkeys of approximately 50% may be too low for the testing of sporozoite vaccines or drugs directed against the exoerythrocytic stages. However, the strain is highly infectious to commonly available laboratory-maintained anopheline mosquitoes. Mosquito infection is especially high when feedings are made with gametocytes from splenectomized chimpanzees.

Trypanosoma cruzi infection in Leontopithecus rosalia at the Reserva Biológica de Poco das Antas, Rio de Janeiro, Brazil.

Wild golden lion tamarins (Leontopithecus rosalia) - endangered primates that are native to the Brazilian Atlantic coastal forest - were surveyed for the presence of Trypanosoma cruzi with the use of Giemsa-stained blood smears, hemocultures and an indirect immunofluorescence assay (IFAT). Positive IFAT with titers ranging from 1:20 to 1:1280 were observed in 52% of the 118 wild tamarins examined and the parasite was isolated from 38 tamarins. No patent parasitemia was observed among the tamarins from which T. cruzi was isolated. Serum conversion and positive hemoculture was observed for three animals that had yielded negative results some months earlier, which indicates that T. cruzi is actively transmitted among tamarins. In contrast to observations with other sylvatic isolates, those from the tamarins were significantly more virulent and most of them produced mortality in experimentally infected Swiss mice. Some variation in the kDNA restriction profiles among the isolates was observed. Electrophoresis with GPI, G6PDH, IDH, MDH and ME enzymes showed a Z2 profile.

Trypanosoma cruzi infection in Leontopithecus rosalia at the Reserva Biológica de Poço das Antas, Rio de Janeiro, Brazil

Wild golden lion tamarins (Leontopithecus rosalia) -- endangered primates that are native to the Brazilian Atlantic coastal forest -- were surveyed for the presence of Trypanosoma cruzi with the use of Giemsa-stained blood smears, hemocultures and an indirect immunofluorescence assay (IFAT). Positive IFAT with titers ranging from 1:20 to 1:1280 were observed in 52 per cent of the 118 wild tamarins examined and the parasite was isolated from 38 tamarins. No patent parasitemia was observed among the tamarins from which T. cruzi was isolated. Serum conversion and positive hemoculture was observed for three animals that had yielded negative results some months earlier, which indicates that T. cruzi is actively transmitted among tamarins. In contrast to observations with other sylvatic isolates, those from the tamarins were significantly more virulent and most of them produced mortality in experimentally infected Swiss mice. Some variation in the kDNA restriction profiles among the isolates was observed. Electrophoresis with GPI, G6PDH, IDH, MDH and ME enzymes showed a Z2 profile.

Monkeys of the rainforest in French Guiana are natural reservoirs for P. brasilianum/P. malariae malaria.

Monkey blood samples were collected from 214 monkeys relocated as part of the wildlife rescue organized in French Guiana during the filling of the Petit Saut Dam on the Sinnamary River. These samples were tested for malaria parasites by microscopy of thick blood filsm and by nested PCR for small subunit rRNA genes (SSUrRNA). Parasitic blood forms similar to Plasmodium brasilianum were detected in 4 monkey species: Alouatta seniculus macconnelli, Saguinus midas midas, Pithecia pithecia and Ateles paniscus paniscus, with the highest prevalence in Alouatta monkeys. PCR was more sensitive than the conventional method for detecting low-grade parasitaemia in positive monkeys. The examination of blood films indicated that 5.6% of the animals carried parasites whereas the nested PCR for ribosomal DNA indicated a prevalence of 11.3%. The P. brasilianum SSUrRNA gene sequence was analysed and aligned with those from P. malariae, P. falciparum and P. vivax. This suggested that P. brasilianum and P. malariae are very closely related. Similar results were obtained from analysis of the sequences in P. malariae and P. brasilianum isolates of a polymorphic gene fragment analogous to the merozoite surface protein-1 (MSP-1) gene of P. falciparum. The P. brasilianum/P. malariae sequences were more similar to those of P. vivax than to those of P. falciparum, at least in the gene region examined. The high degree of DNA homology in the sequences of the SSUrRNA and msp1-like genes is consistent with other characterizations demonstrating a taxonomic relationship between P. brasilianum and P. malariae species. Our results provide further evidence that P. brasilianum and P. malariae are virtually identical and should probably be considered to be a single malaria species. This raises the question as to whether monkeys living in the rainforest are natural reservoirs for both simian and human malaria. These results have implications for the interpretation of the current epidemiological situation in French Guiana.

Presumptive dirofilariasis in a pale-headed saki monkey (Pithecia pithecia).

A 6-yr-old male pale-headed saki monkey (Pithecia pithecia), born at the Dallas Zoo, reentered the collection in 1994 after it was housed for 4 yr in Rhode Island and 2 yr in Florida. The monkey tested negative for both Dirofilaria immitis microfilariae and D. immitis adult antigens (via commercially available tests) upon return. However, it tested positive for adult antigens 1 yr later, and additional testing, including ultrasonography, suggested a diagnosis of aberrant dirofilariasis. Relevant evidence of previous microfilaremia in pale-headed saki monkeys at the Dallas Zoo is reviewed. Dirofilaria immitis infection should be included in the differential diagnosis list for any nonhuman primate with cardiopulmonary disease wherever the parasite is enzootic.

[Parasitological evidence on the phylogeny of hominids and cebids]. / Evidencia parasitológica sobre la filogenia de los homínidos y los cébidos.

A systematic revision of the ectoparasites (lice) of the hominids and ceboids supports the Trogloditian hypothesis, according to which the genus Homo is the sister of Pan, and the genus Gorilla the sister group of both. The phylogenetic analysis of this matrix derived from the study of primate lice shows an C.I. of 0.71 for the Trogloditian hypothesis including the ceboids in the analysis.

Evidencia parasitológica sobre la filogenia de los homínidos y los cébidos / Parasitological evidence about the phylogeny of the hominoids and the ceboids

A systematic revision of the ectoparasites (lice) of the hominids and ceboids supports the Trogloditian hypothesis, according to which the genus Homo is the sister of Pan, and the genus Gorilla the sister group of both. The phylogenetic analysis of this matrix derived from the study of primate lice shows an C.I. of 0.71 for the Trogloditian hypothesis including the ceboids in the analysis

Simian malaria at two sites in the Brazilian Amazon. I--The infection rates of Plasmodium brasilianum in non-human primates.

The parasite that causes simian malaria in the Brazilian Amazon, Plasmodium brasilianum, is infective to man. In this region, where humans live within and in close proximity to the forest, it was suspected that this parasite could be the cause of a zoonosis. A study was performed in the areas surrounding two hydroelectric plants in the Amazon, Balbina and Samuel, aiming at determining the zoonotic potential of this parasite. P. brasilianum was detected in, respectively, 15.8% and 9.9% of 126 and 252 primates belonging to seven and eight species examined from Balbina and Samuel. The highest malaria infection rates were found among the red-howler monkey Alouatta seniculus straminea (32.3%), the bearded-saki Chiropotes satanas chiropotes (50%) and the spider-monkey Ateles paniscus paniscus (2[1+]) from Balbina and in the squirrel-monkey Saimiri ustus (21%) and the black-faced-spider-monkey Ateles paniscus chamek (28.6%) from Samuel.

Intestinal capillariasis in New World monkeys.

Two cases of intestinal capillariasis have been identified at necropsy in a squirrel monkey and a capuchin monkey born and raised in captivity. The parasites are described as far as possible from the histopathological slides or intestinal contents, and their relationship to other intestinal capillarids, especially those of primates, is discussed.

Simian malaria in Brazil.

In Brazil simian malaria is widely spread, being frequent in the Amazon region (10% of primates infected) and even more in the forested coastal mountains of the Southeastern and Southern regions (35% and 18% infected, respectively), but absent in the semi-arid Northeast. Only two species of plasmodia have been found: the quartan-like Plasmodium brasilianum and the tertian-like P. simium, but the possible presence of other species is not excluded. P. brasilianum is found in all enzootic foci, but P. simium was detected only on the coast of the Southeastern and Southern regions, between paralles 20 degrees S and 30 degrees S. Nearly all hosts are monkeys (family Cebidae, 28 species harbouring plasmodia out of 46 examined), and very rarely marmosets or tamarins (family Callitrichidae, 1 especies out of 16). P. brasilianum was present in all infected species, P. simium in only two. The natural vector in the Southeastern and Southern regions was found to be Anopheles cruzi, but has not been conclusively identified in the Amazon. One natural, accidental human infection due to P. simium was observed. There is no evidence of the relation of simian to human malaria in the Southeastern and Southern regions, where human malaria was eradicated in spite of the high rates of monkeys infected, but in the Amazon recent serological studies by other workers, revealing high positivity for P. brasilianum/P. malariae antibodies in local indians, would suggest that among them malaria might possibly be regarded as a zoonosis.

Simian malaria in Brazil

In Brazil simian malaria is widely spread, being frequent in the Amazon region (10% of primates infected) and even more in the forested coastal mountains of the Southeastern and Southern regions (35% and 18% infected, respectively), but absent in the semi-arid Northeast. Only two species of plasmoidia have been found: the quartan-like Plasmodium brasilianum and the tertian-like P. simium, but the possible presence of other species is not excluded. P. brasilianum is found in all enzootic foci, but P. simium was detected only on the coast of the Southeastern and Southern regions, between parallels 20-S and 30-S. Nearly all hosts are monkeys (family Cebidae, 28 species harbouring plasmodia out of 46 examined) and very rarely marmosets or tamarins (family Callitrichidae, I especies out of 16). P. brasilianum was present in all infected species, P. simium in only two. The natural vector in the Southeastern and Southern regions was found to be Anopheles cruzi, but has not been conclusively identified in the Amazon. One natural, accidental human infection due to P. simium was observed. There is no evidence of the relation of the simian to human malaria in the Southeastern and Southern regions, where human malaria was eradicated in spite of the high rates of monkeys infected, but in the Amazon recent serological studies by other workers, revealing high positivity for P. brasilianum/P. malariae antibodies in local indians, would suggest that among them malaria might be regarded as a zoonosis

Protection of aouts monkeys after immunization with recombinant antigens of Plasmodium falciparum

The genus Aotus spp. (owl monkey) is one of the WHO recommended experimental models for Plasmodium falciparum blood stage infection, especially relevant for vaccination studies with asexual blood stage antigens of this parasite. For several immunization trials with purified recombinant merozoite/schizont antigens, the susceptible Aouts kenotypes II, III, IV and VI were immunized with Escherichia coli derived fusion proteins containg partial sequences of the proteins MSAI (merozoite surface antigen I), SERP (serine-strech protein) and HRPII (histidine alanine rich protein II) as well as with a group of recombinant antigens obtained by an antiserum raised against a protective 41 kD protein band. The subcutaneous application (3x) of the antigen preparations was carried out in intact animals followed by splenectomy prior to challange, in order to increase the susceptibility of the experimental hosts to the parasite. A partial sequence of HRPII, the combination of three different fusion proteins of the 41 kD group and mixture of two sequences of SERP in the presence of the modified Al(OH)3 adjuvant conferred significant protection against a challange infection with P. falciparum blood stages (2-5 x 10 (elevado a sexta potência) i. RBC). Monkey immunized with the MS2-fusion protein carrying the N-terminal part of the 195 kD precursor of the major merozoite surface antigens induced only marginal protection showing some correlation between antibody titer and degree of parasitaemia. Based on the protective capacity of these recombinant antigens we have expressed two hybrid proteins (MS2/SERP/HRPII and SERP/MSAI/HRPII) in E. coli containing selected partial sequences of SERP, HRPII and MSAI. Antibodies raised against both hybrid proteins in rabbits and Aotus monkeys recognize the corresponding schizont polypeptides. In two independent immunization trials using 13 animals (age 7 months to 3 years) we could show that immunization of Aotus monkeys with either of the two hybrid proteins administrated in an oil-based well tolerated formulation protected the animals from severe experimental P. falciparum (strain Palo Alto) infection

Efficience of human Plasmodium falciparum malaria vaccine candidates in Aotus lemurinus monkeys

The protective efficacy of several recombinat and a synthetic Plasmodium falciparum protein was assessed in Aoutus monkeys. The rp41 aldolase, the 190L fragment of the MSA-1 protein and fusion 190L-CS. T3 protein containg the CS. T3 helper "universal epitope were emulsified in Freund's adjuvants and injected 3 times in groups of 4-5 monkeys each one. The synthetic polymer Spf (66)30 also emulsified in Freund's adjuvants was injected 6 times. Control groups for both experiments were immunized with saline solution in the same adjuvant following the same schedules. Serology for malaria specific antibodies showed seroconversion in monkeys immunized with the recombinant proteins but not in those immunized with the polymer nor in the controls. Challenge was performed with the 10 (elevado a quinta potência) parasites from the P. falciparum FVO isolate. Neither rp41 nor SPf (66)30 induced protection, whereas 190L induced significant delay of parasitemia. The fusion of the CS. T3 epitope to 190L significantly increased is protective capacity