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.

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.

In vitro and in vivo adaptation of the Geneve/SGE-1 strain of Plasmodium falciparum to growth in a squirrel monkey (Saimiri sciureus) model.

Human erythrocytic culture-adapted parasites of the Geneve/SGE-1 strain of Plasmodium falciparum were successfully adapted to grow in an in vitro culture system containing squirrel monkey erythrocytes and serum. These monkey culture-adapted organisms were then used to produce a patent infection in a splenectomized squirrel monkey. Fresh infected blood from this animal was introduced into another splenectomized monkey and was subsequently serially passed through seven intact squirrel monkeys. High level parasitemias (greater than 10%) were obtained in the animals from the last two passes following inoculation of moderate numbers of parasites. It is anticipated that this squirrel monkey-adapted Geneve/SGE-1 strain of P. falciparum will continue to produce high level parasitemias in intact Bolivian Saimiri, and consequently will be suitable for challenge of these monkeys.

Adaptation of the Panama II strain of Plasmodium falciparum to Panamanian owl monkeys.

The Panama II strain of Plasmodium falciparum, acquired at the second passage level in splenectomized Colombian owl monkeys, was adapted to owl monkeys of Panamanian origin. Patent infections were induced in 22 of 27 unaltered and 20 of 21 splenectomized recipients during 19 serial passages. The infections were significantly more virulent in splenectomized than normal Panamanian owl monkeys, however recrudescences in seven normal monkeys achieved peak parasitemias 48 times greater than in the primary attack. These results describe the first reproducible infections of indigenous falciparum malaria in Panamanian owl monkeys.

Plasmodium inui and Babesia microti infections in the squirrel monkey, Saimiri sciureus.

The course of Plasmodium inui and Babesia microti infections was studied in seven splenectomized squirrel monkeys (Saimiri sciureus) of Guyanan or Bolivian origin. Three of the monkeys were infected with P. inui either by the inoculations of parasitized blood or by the bite of infected mosquitoes. The remaining four monkeys were infected by the inoculation of parasitized blood, containing P. inui and B. microti in three and with B. microti only in one. The infection in all seven animals was severe, terminating fatally.

Adaptation of a Nigerian strain of Plasmodium falciparum to Panamanian Aotus trivirgatus.

A patent infection (more than 190 days duration) with Plasmodium falciparum, obtained directly from a patient who had been exposed to malaria in Nigeria, was established in a splenectomized Panamanian owl monkey, Aotus trivirgatus griseimembra. Subinoculations from the original monkey recipient into one splenectomized and two normal A. t. griseimembra produced primary patient parasitemias of 18-54 days duration. These results represent the first successful adaptation of P. falciparum from man to a monkey of Panamanian origin.

Susceptibility of Peruvian Aotus monkeys to infection with different species of Plasmodium.

Splenectomized Aotus trivirgatus monkeys of Peruvian origin were tested for their susceptibility to infection with different species of Plasmodium. Strains of P. vivax from Vietnam and El Salvador produced infections in eight animals with maximum parasite densities ranging from 2,000-180,000 per mm3. A strain of P. falciparum from Cambodia produced maximum parasitemias in five animals ranging from 8-25% of the red blood cells infected. Although two monkeys inoculated with P. fragile had maximum parasitemias of 31% and 42%, treatment was not necessary for animal survival. Three monkeys infected with P. knowlesi via the bites of infected mosquitoes had maximum parasitemias of from 25-41%; these animals died of their infections. Nine animals were infected with P. cynomolgi; maximum parasitemias in seven of these animals with no prior history of malarial infection ranged from 49,500 per mm3 to 11%; two animals with prior experience with P. vivax and P. fragile had maximum P. cynomolgi parasitemias of 65 and 13,200 per mm3. No infections were obtained in animals inoculated with P. coatneyi or P. hylobati.

Pathologic changes associated with fatal Plasmodium falciparum infection in the Bolivian squirrel monkey (Saimiri sciureus boliviensis).

Fatal cases of experimental Plasmodium falciparum (Indochina I) in Bolivian squirrel monkeys (Saimiri sciureus boliviensis) were examined by histologic and ultrastructural methods. Gross lesions were characterized by hepatosplenomegaly and interstitial pulmonary changes. Histologically, there was marked diffuse reticuloendothelial hyperplasia, pulmonary alveolar septal thickening, mesangioproliferative glomerulonephropathy, sequestration of parasitized erythrocytes in deep vascular beds, degenerative parenchymal changes in the liver and myocardium, and in one case retinal and cerebral hemorrhage. These data indicate that the Bolivian squirrel monkey is a good model for studying pathologic changes associated with human falciparum malaria.

Susceptibility of different populations of Colombian Aotus monkeys to the FCB-1 strain of Plasmodium falciparum.

p6udies of susceptibility to the FCB-1 strain of Plasmodium falciparum from Colombia were conducted in three different groups of Aotus monkeys. The results indicated that Aotus trivirgatus griseimembra from the north of the Magdalena Valley were the most susceptible host. A. t. griseimembra from the central Magdalena Valley appeared to be less susceptible to infection, while Aotus monkeys from the eastern Andes were even more resistant to infection. Serum electrophoretic studies were not useful as indicators of resistance or susceptibility to infection. However, the presence of 58 chromosomes in monkeys from the east Andes, a karyotype not previously reported, may be an indicator for the resistance of these animals to the FCB-1 strain. Based on these results, it is possible to assume that Aotus monkeys from the eastern Andes region of Colombia may represent a new species of Aotus; further studies are needed to clarify its taxonomic status.

Susceptibility of Aotus trivirgatus to Leishmania braziliensis and L. mexicana.

Two Aotus trivirgatus (owl monkeys) were infected experimentally with Leishmania braziliensis and two with L. mexicana strains of Panamanian origin in a pilot study to determine the susceptibility and the course of infection of cutaneous leishmaniasis in this primate species. Montenegro skin tests performed on all animals prior to parasite inoculation were negative. A standardized inoculum of promastigotes was injected intradermally on the nose of each monkey. All of the animals developed infections which lasted from 3.5 to 8.5 months. Depigmentation developed at the site of the inoculation in all of the subjects. The severity of the resulting lesions was greater in the animals infected with L. braziliensis. Positive skin tests developed in three A. trivirgatus at days 62, 76, and 139 postinoculation, respectively. An explanation for the negative skin test in the fourth animal is discussed.

Comparison of Plasmodium falciparum infections in Panamanian and Colombian owl monkeys.

Parameters of blood-induced infections of the Vietnam Oak Knoll, Vietnam Smith, and Uganda Palo Alto strains of Plasmodium falciparum studied in 395 Panamanian owl monkeys in this laboratory between 1976-1984 were compared with those reported from another laboratory for 665 Colombian owl monkeys, studied between 1968-1975, and, at the time, designated Aotus trivirgatus griseimembra. The virulence of these strains was less in Panamanian than in Colombian owl monkeys, as indicated by lower mortality rates of the Panamanian monkeys during the first 30 days of patency. Maximum parasitemias of the Vietnam Smith and Uganda Palo Alto strain, in Panamanian owl monkeys dying during the first 15 days of patent infection, were significantly higher than in Colombian owl monkeys. Panamanian owl monkeys that survived the primary attack had significantly higher maximum parasitemias than the surviving Colombian owl monkeys. Peak parasitemias were attained significantly earlier after patency in Panamanian than in Colombian owl monkeys, irrespective of the strain of P. falciparum. More Panamanian than Colombian owl monkeys evidenced self-limited infection after the primary attack of either the Vietnam Smith or Uganda Palo Alto strain. The duration of the primary attacks and recrudescences were significantly shorter in Panamanian than in Colombian owl monkeys. Mean peak parasitemias during recrudescence were usually higher in Panamanian owl monkeys than in Colombian monkeys. Differences of infection parameters were probably attributable, in part, to geographical origin of the two monkey hosts and parasite strains.

Adaptation of the Indochina I/CDC strain of Plasmodium falciparum to the squirrel monkey (Saimiri sciureus).

The Indochina I/CDC strain of Plasmodium falciparum was linearly passaged in squirrel monkeys (Saimiri sciureus) of 3 phenotypes. Splenectomized monkeys of Guyanan and Peruvian type developed high density parasitemias, but considerably lower than the mean peak parasitemia (greater than 10(6)/mm3) in Bolivian phenotype squirrel monkeys. Spleen-intact Bolivian and Peruvian squirrel monkeys all developed potentially lethal infections after linear passage of parasites from Saimiri and Aotus. For the evaluation of induced immunity to P. falciparum, the Indochina I/CDC strain in Saimiri will be a valuable model system.

Eosinophilic meningoencephalitis due to Angiostrongylus cantonensis as the cause of death in captive non-human primates.

Fatal eosinophilic meningoencephalitis due to Angiostrongylus cantonensis is reported in captive non-human primates. A howler monkey (Alouatta caraya) at the Audubon Park and Zoological Gardens, New Orleans, LA, died 21 days after initial clinical symptoms. A white-handed gibbon (Hylobates lar) died at the Ardastra Gardens and Zoo, Nassau, Bahamas, 17 days after onset of symptoms. Both had access to free-ranging gastropods within the zoos. These are the first reported cases of natural infection by A. cantonensis in non-human primates in the western hemisphere.

Invasive amebiasis in naturally infected New World and Old World monkeys with and without clinical disease.

Histopathological preparations of cecum and colon from monkeys naturally infected with invasive Entamoeba histolytica were examined to determine the distribution of amebae in the tissues and the types of lesions, if any, associated with them. Infections were studied in 3 New World species (10 Callicebus moloch, 1 C. torquatus, and 2 Aotus trivirgatus) and 3 Old World species (8 Macaca mulatta, 6 Erythrocebus patas, and 1 Cercopithecus aethiops). Amebiasis was recorded as the principal or a contributing cause of death of all of the 13 New World monkeys and in 6 of the 15 Old World monkeys; amebiasis was detected in the rest of the monkeys only after tissues were re-examined specifically for amebae. Amebae causing no apparent damage were found in the lamina propriae, mainly at the muscularis mucosae. Most frequent were colonies or aggregates of amebae in the crypts between the epithelium and basement membrane, causing either no evident necrosis or changes ranging from necrosis and disarrangement of adjacent cells to complete destruction of the epithelium and reduction of the cells to pyknotic bodies. A lesion interpreted as possibly characteristic of carrier-state invasive amebiasis was destruction of the epithelium in patches of mucosal crypts, not leading to ulceration. Uncommon but present in both New and Old World monkeys were typical areas of surface erosion and classical flask-shaped ulcers. The observations show that in some species of Old World monkeys amebiasis can be invasive without causing clinical disease.

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.

Attempts to establish Onchocerca volvulus infection in primates and small laboratory animals.

Attempts were made to transmit O. volvulus infection to small laboratory animals and several species of primates to identify a practical laboratory host for human onchocerciasis. Infective larvae of O. volvulus of Guatemalan origin were inoculated into the following animals: rhesus monkeys, bonnet monkeys, golden spider monkeys, black spider monkeys, galagos, opossums, jirds, newborn and adult Swiss mice, kinkajou, cebus monkey, normal and splenectomized multimammate rats, and a calf. The animals were examined for signs of developing infection for up to two years after inoculation. None of the animals tested developed a patent infection, and neither larvae nor O. volvulus adults were found during the necropsy of all the animals which died or were killed during or at the end of the examination period. It is concluded that none of the animals tested is susceptible to infection with O. volvulus.

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.

Infection of Aotus azarae boliviensis monkeys with different strains of Plasmodium vivax.

Sixty-seven splenectomized Aotus azarae boliviensis were infected with strains of Plasmodium vivax from Southeast Asia (2), New Guinea (2), North Korea (1), and Central America (3). Maximum parasitemias varied among the different strains, with the mean maximum parasitemia for the primary infection period being 16,200 per mm3. Animals previously infected with Plasmodium falciparum and Plasmodium malariae produced maximum parasitemias of 30,200 and 11,900 per mm3, respectively. Gametocytes infective to Anopheles freeborni mosquitoes were produced with 7 of the 8 strains examined.

Infection of Peruvian Aotus nancymai monkeys with different strains of Plasmodium falciparum, P. vivax, and P. malariae.

Aotus nancymai (karyotype I) monkeys from Peru were studied for their susceptibility to infection with Plasmodium falciparum, P. vivax, and P. malariae. Three strains of P. falciparum (Santa Lucia from El Salvador, Indochina I/CDC from Thailand, and Uganda Palo Alto) were inoculated into 38 monkeys. The results indicated that this species of Aotus monkey is highly susceptible to infection. The Uganda Palo Alto and the Santa Lucia strain parasites appear to be the most useful for immunologic and chemotherapeutic studies. Five strains of P. vivax (Chesson, ONG, Vietnam Palo Alto, Salvador I, and Honduran I/CDC) were inoculated into 28 monkeys. The Vietnam Palo Alto strain produced the highest level parasitemias ranging from 23,800 to 157,000/mm3. Mosquito infections were obtained with the ONG, Chesson, and Salvador I strains. Two out of 6 attempts to transmit P. vivax via sporozoite inoculation to splenectomized monkeys were successful with prepatent periods of 39 and 57 days. Five monkeys were infected with the Uganda I/CDC strain of P. malariae. Maximum parasitemias ranged from 10 to 5,390/mm3.

Transmission of the OS strain of Plasmodium inui to Saimiri sciureus boliviensis and Aotus azarae boliviensis monkeys by Anopheles dirus mosquitoes.

Eight Saimiri and 7 Aotus monkeys were exposed to infection with the OS strain of Plasmodium inui via the bites of from 2 to 7 Anopheles dirus mosquitoes. All Saimiri monkeys developed high-level infections of from 152,000 to 500,000/mm3 after prepatent periods of from 14 to 17 days. Only 1 Aotus monkey developed a patent infection after a period of 28 days. Feeding on these animals failed to result in infection of An. dirus mosquitoes.