Occurrence of Free-Living Amoebae in Non-Human Primate Gut.

The gut microbiome reflects health and predicts possible disease in hosts. A holistic view of this community is needed, focusing on identifying species and dissecting how species interact with their host and each other, regardless of whether their presence is beneficial, inconsequential, or detrimental. The distribution of gut-associated eukaryotes within and across non-human primates is likely driven by host behavior and ecology. To ascertain the existence of free-living amoebae (FLA) in the gut of wild and captive non-human primates, 101 stool samples were collected and submitted to culture-dependent microscopy examination and DNA sequencing. Free-living amoebae were detected in 45.4% (46/101) of fecal samples analyzed, and their morphological characteristics matched those of Acanthamoeba spp., Vermamoeba spp., heterolobosean amoeboflagellates and fan-shaped amoebae of the family Vannellidae. Sequence analysis of the PCR products revealed that the suspected amoebae are highly homologous (99% identity and 100% query coverage) with Acanthamoeba T4 genotype and Vermamoeba vermiformis amoebae. The results showed a great diversity of amoebae in the non-human primate's microbiome, which may pose a potential risk to the health of NHPs. To our knowledge, this is the first report of free-living amoebae in non-human primates that are naturally infected. However, it is unknown whether gut-borne amoebae exploit a viable ecological niche or are simply transient residents in the gut.

Lessons from a Multilaboratorial Task Force for Diagnosis of a Fatal Toxoplasmosis Outbreak in Captive Primates in Brazil.

Toxoplasmosis is an important zoonotic disease caused by the parasite Toxoplasma gondii and is especially fatal for neotropical primates. In Brazil, the Ministry of Health is responsible for national epizootic surveillance, but some diseases are still neglected. Here, we present an integrated investigation of an outbreak that occurred during the first year of the COVID-19 pandemic among eleven neotropical primates housed at a primatology center in Brazil. After presenting non-specific clinical signs, all animals died within four days. A wide range of pathogens were evaluated, and we successfully identified T. gondii as the causative agent within four days after necropsies. The liver was the most affected organ, presenting hemorrhage and hepatocellular necrosis. Tachyzoites and bradyzoite cysts were observed in histological examinations and immunohistochemistry in different organs; in addition, parasitic DNA was detected through PCR in blood samples from all specimens evaluated. A high prevalence of Escherichia coli was also observed, indicating sepsis. This case highlights some of the obstacles faced by the current Brazilian surveillance system. A diagnosis was obtained through the integrated action of researchers since investigation for toxoplasmosis is currently absent in national guidelines. An interdisciplinary investigation could be a possible model for future epizootic investigations in animals.

Isolation and genetic characterization of Toxoplasma gondii from a captive black-and-gold howler monkey (Alouatta caraya Humboldt, 1812) in Brazil.

Toxoplasma gondii was isolated in mice from different tissues of a captive black-and-gold howler monkey (Alouatta caraya) kept in a colony at the Primatology Center of Rio de Janeiro State, Brazil, and it was genotypically characterized based on using PCR-RFLP and Microsatellite Analysis (MS), later on. T. gondii was successfully isolated from inocula deriving from heart, liver and tissue pool (heart, liver, lungs, axillary lymph nodes and cerebellum) samples. The isolate was named TgBgHmBrRJ1. The high virulence of the aforementioned strain was observed in infected mice. Non-archetypal genotype (ToxoDB PCR-RFLP #206) was obtained through PCR-RFLP. This genotype had been previously described in 12 isolates from different hosts, also in Southeastern Brazil, a fact that indicates likely high circulation of this genotype in this region. The isolate was also classified as non-archetypal, based on MS genotyping, as well as presented genotypic identity close to that of strains isolated from free-range non-symptomatic chickens (TgCkBr244,245,278,279) in Espírito Santo State. It is worth emphasizing that despite the large number of reports about clinical toxoplasmosis in neotropical primates in Brazil, this is just the second isolate of this parasite ever reported in this group of animals.

Pulmonary Granuloma Is Not Always the Tuberculosis Hallmark: Pathology of Tuberculosis Stages in New World and Old World Monkeys Naturally Infected with the Mycobacterium tuberculosis Complex.

We present the pathology of monkeys naturally infected with Mycobacterium tuberculosis complex from five different colonies in Rio de Janeiro, Brazil. On the basis of gross and histopathological findings, the lesions were classified into chronic-active, extrapulmonary, early-activation or latent-reactivation stages. Typical granulomatous pneumonia was seen in 46.6% of cases (six rhesus monkeys [Macaca mulatta] and one Uta Hick's bearded saki [Chiropotes utahickae]). The absence of pulmonary granulomas did not preclude a diagnosis of tuberculosis (TB): classical granulomatous pneumonia was observed in the chronic-active and latent-reactivation stages but not in the extrapulmonary and early-activation stages. The early-activation stage was characterized by interstitial pneumonia with a predominance of foamy macrophages and molecular and immunohistochemical evidence of M. tuberculosis complex infection. TB should be considered as a cause of interstitial pneumonia in New World Monkeys. We recommend the use of immunohistochemistry and molecular analysis for diagnosis of TB, even when typical macroscopic or histological changes are not observed.

Subacute multisystemic toxoplasmosis in a captive black-and-gold howler monkey (Alouatta caraya) indicates therapy challenging.

A 10-year-old black howler monkey presented with a 36-day subacute clinicopathological picture of fever, prostration, inappetence, intestinal hypomotility, and emaciation. Therapy was trimethoprim-sulfamethoxazole with streptomycin. The liver, lungs, lymph nodes, and spleen presented lesions. Toxoplasma gondii isolation and PCR determined the diagnosis, and indirect fluorescent antibody tests confirmed an increase in antibody titers.

Ultrasound monitoring of the uterus and ovaries of dominant and subordinate females of yellow-breasted capuchin (Sapajus xanthosternos) and robust tufted capuchin (Sapajus robustus) in captive colonies during the ovarian cycle and anestrus periods / Acompanhamento ultrassonográfico do útero e ovários de fêmeas dominantes e subordinadas de macacos-prego-do-peito-amarelo (Sapajus xanthosternos) e macacos-prego-de-crista (Sapajus robustus) em colônias cativas, durante o ciclo ovariano e períodos de anestro

The yellow-breasted capuchin (Sapajus xanthosternos) and robust tufted capuchin (Sapajus robustus) are endangered species due to destruction of their natural habitat and predatory chase. However, it is still necessary to elucidate some details of their reproductive physiology in order to obtain better indices in the assisted reproduction of these species. This study aimed to evaluate the ovarian cycle of 13 dominant and subordinate females of S. xanthosternos (n=8) and S. robustus (n=5) using sagittal and transversally scanned ultrasound of their uterus and ovaries. Sonograms were performed every seven days for two months. The ovarian cycle phase and anestrous condition were confirmed by colpocytology. Our results showed different uterine parameters (craniocaudal diameter, dorso-ventral diameter, and transverse diameter) (P<0.05) between anestrous subordinate females and other ovarian cycle phases and social classes. The mean of uterine volume was higher in dominant females than subordinate females in all cycle phases (P<0.05), except in follicular phase. During anestrus, endometrial width was smaller in subordinate females than in dominant females (P<0.05). Subordinate females showed differences in endometrial measures (P<0.05) between anestrous period and follicular and luteal periods. Ovarian measures in dominant females were higher than in subordinate females only during anestrus (P<0.05). In the subordinate females, ovarian parameters were different (P<0.05) between anestrus and follicular and luteal phases. Dominant females showed higher volume of right ovary compared to volume of the left ovary during anestrus and follicular phase (P<0.05). Follicles and corpus luteum were distinguished by ultrasonography in most exams (86.11%). During anestrus, measurable ovarian structures were not observed in both ovaries in dominant and subordinate females. In conclusion, the methodology used in this study allowed to evaluate the ovarian cycle in S. xanthosternos e S. robustus females and that cycle phase/anestrus and social class of the female influenced the size of the uterus and ovaries.(AU)

O macaco-prego-do-peito-amarelo (Sapajus xanthosternos) e o macaco-prego-de-crista (Sapajus robustus) encontram-se em risco de extinção devido a destruição do seu habitat e a caça predatória. Porém, ainda necessita-se elucidar alguns detalhes de sua fisiologia reprodutiva, para obterem-se melhores índices por meio de reprodução assistida. Sendo assim, o objetivo deste trabalho foi acompanhar o ciclo ovariano de 13 fêmeas dominantes e subordinadas de S. xanthosternos (n=8) e S. robustus (n=5) por meio de cortes ultrassonográficos sagitais e transversais do útero e dos ovários. Estas fêmeas foram examinadas uma vez por semana durante dois meses. A fase do ciclo ovariano/anestro foi confirmada pela colpocitologia. Os resultados da pesquisa demonstrou diferença significativa (P<0,05) relacionada ao diâmetro crânio-caudal, diâmetro dorso-ventral e diâmetro transversal entre as fêmeas subordinadas em anestro com todas as outras fases do ciclo ovariano e classes sociais. A média do volume uterino foi maior nas fêmeas dominantes que nas subordinadas em todas as fases do ciclo (P<0,05) a exceção da fase folicular. A largura endometrial, durante o período de anestro, foi menor nas fêmeas subordinadas quando comparada às dominantes (P<0,05). Nas fêmeas subordinadas, houve diferenças (P<0,05) nas mensurações endometriais entre o período de anestro e das fases folicular e lútea. Quanto aos ovários, as médias das medidas observadas nas fêmeas dominantes foram superiores as das subordinadas durante o anestro (P<0,05). As médias das medidas das variáveis ovarianas das fêmeas subordinadas apresentaram diferenças entre o anestro e as fases folicular e lútea (P<0,05). Nas fêmeas dominantes o volume do ovário direito foi maior que o do ovário esquerdo durante o anestro e na fase folicular (P<0,05). Folículos e corpos lúteos foram diferenciados pela ultrassonografia na maioria das coletas (86,11%). Durante o anestro não foram observadas estruturas ovarianas mensuráveis nos ovários em ambas as classes sociais. Concluiu-se que a metodologia empregada neste estudo permitiu o acompanhamento do ciclo ovariano das fêmeas S. xanthosternos e S. robustus e que as fases do ciclo/anestro e a classe social das fêmeas influenciaram as medidas do útero e ovários.(AU)

Detection of mycobacterial infection in non-human primates using the Xpert MTB/RIF molecular assay.

Tuberculosis is a major public health concern, and diagnostic strategies applied to animal populations are scarce. As part of ongoing efforts to control tuberculosis dissemination at our animal facility, two non-human primates (NHP, Saimiri sciureus) presenting cutaneous lesions were examined for mycobacterial infection. Both animals tested positive for acid-fast bacilli and Mycobacterium tuberculosis using a molecular assay (IS6110 PCR). Animals were euthanized and several samples were tested for M. tuberculosis using the Xpert MTB/RIF assay. Many samples were positive for M. tuberculosis and rifampicin resistance, and some produced mycobacterial growth. Oral swabs from cage mates were then tested with Xpert MTB/RIF, and the majority tested positive for M. tuberculosis and rifampicin resistance, and produced growth in culture. To our knowledge, this is the first report of multidrug-resistant mycobacterial infection in NHP. Additionally, our data shows that the Xpert MTB/RIF assay can be useful as a screening tool for tuberculosis infection in NHP.

Myocardial stereology in captive Callithrix kuhlii (Callitrichidae, Primates): healthy animals versus animals affected by wasting marmoset syndrome (WMS)

This study comprised 12 hearts of Wied´s black-tufted-ear marmoset, Callithrix kuhlii (Coimbra-Filho 1985), 6 with Wasting Marmoset Syndrome (WMS) and 6 non-affected. Biometry was performed after death. After necropsy, the hearts were weighed, dissected, fixed in 10 percent formalin solution (pH 7.2), and processed for optical microscopy at 5µm sections stained with Haematoxylin-Eosin. Quantitative analysis was performed by stereological techniques. The statistical differences between the biometrical and stereological parameters were assessed by the Mann-Whitney test. The morphometric results showed that WMS causes a significant reduction in body and cardiac weights, and also in the volume density of vessels in those animals. Further studies are necessary to understand some of the results shown here.

Neste estudo, foram utilizados corações de 12 Sagui-de-Wied, Callitrhix kuhlii (Coimbra-Filho 1985), sendo 6 animais afetados pela SEP e 6 animais normais. Após a morte foi realizada a biometria seguida de necropsia. Os corações foram fixados em formol tamponado a 10 por cento, pesados e dissecados, sendo processados através de técnicas histológicas de rotina para microscopia óptica em cortes de 5µm corados por Hematoxilina-Eosina. As análises quantitativas foram feitas com o uso de técnicas estereológicas. As diferenças estatísticas entre os parâmetros biométricos e estereológicos foram avaliadas usando o test Mann-Whitney. Os resultados encontrados através da morfometria mostraram que a SEP causa uma redução significante do peso tanto corporal quanto do músculo cardíaco, e também uma redução no volume dos vasos nestes animais. Novos estudos são necessários para entender alguns dos resultados mostrados aqui.