The utility of rhesus monkey (Macaca mulatta) and other non-human primate models for preclinical testing of Leishmania candidate vaccines

Leishmaniasis causes significant morbidity and mortality, constituting an important global health problem for which there are few effective drugs. Given the urgent need to identify a safe and effective Leishmania vaccine to help prevent the two million new cases of human leishmaniasis worldwide each year, all reasonable efforts to achieve this goal should be made. This includes the use of animal models that are as close to leishmanial infection in humans as is practical and feasible. Old world monkey species (macaques, baboons, mandrills etc.) have the closest evolutionary relatedness to humans among the approachable animal models. The Asian rhesus macaques (Macaca mulatta) are quite susceptible to leishmanial infection, develop a human-like disease, exhibit antibodies to Leishmania and parasite-specific T-cell mediated immune responses both in vivo and in vitro, and can be protected effectively by vaccination. Results from macaque vaccine studies could also prove useful in guiding the design of human vaccine trials. This review summarizes our current knowledge on this topic and proposes potential approaches that may result in the more effective use of the macaque model to maximize its potential to help the development of an effective vaccine for human leishmaniasis.

Why prevent, diagnose and treat congenital toxoplasmosis?

Evidence that prevention, diagnosis and treatment of toxoplasmosis is beneficial developed as follows: anti-parasitic agents abrogate Toxoplasma gondiitachyzoite growth, preventing destruction of infected, cultured, mammalian cells and cure active infections in experimental animals, including primates. They treat active infections in persons who are immune-compromised, limit destruction of retina by replicating parasites and thereby treat ocular toxoplasmosis and treat active infection in the fetus and infant. Outcomes of untreated congenital toxoplasmosis include adverse ocular and neurologic sequelae described in different countries and decades. Better outcomes are associated with treatment of infected infants throughout their first year of life. Shorter intervals between diagnosis and treatment in utero improve outcomes. A French approach for diagnosis and treatment of congenital toxoplasmosis in the fetus and infant can prevent toxoplasmosis and limit adverse sequelae. In addition, new data demonstrate that this French approach results in favorable outcomes with some early gestation infections. A standardized approach to diagnosis and treatment during gestation has not yet been applied generally in the USA. Nonetheless, a small, similar experience confirms that this French approach is feasible, safe, and results in favorable outcomes in the National Collaborative Chicago-based Congenital Toxoplasmosis Study cohort. Prompt diagnosis, prevention and treatment reduce adverse sequelae of congenital toxoplasmosis.

Circulating natural killer and γδ T cells decrease soon after infection of rhesus macaques with lymphocytic choriomeningitis virus

Rhesus macaques infected with the WE strain of lymphocytic choriomeningitis virus (LCMV-WE) serve as a model for human infection with Lassa fever virus. To identify the earliest events of acute infection, rhesus macaques were monitored immediately after lethal infection for changes in peripheral blood mononuclear cells (PBMCs). Changes in CD3, CD4, CD8 and CD20 subsets did not vary outside the normal fluctuations of these blood cell populations; however, natural killer (NK) and γδ T cells increased slightly on day 1 and then decreased significantly after two days. The NK subsets responsible for the decrease were primarily CD3-CD8+ or CD3-CD16+ and not the NKT (primarily CD3+CD56+) subset. Macaques infected with a non-virulent arenavirus, LCMV-Armstrong, showed a similar drop in circulating NK and γδ T cells, indicating that this is not a pathogenic event. V³9 T cells, representing the majority of circulating γδ T cells in rhesus macaques, displayed significant apoptosis when incubated with LCMV in cell culture; however, the low amount of cell death for virus-co-cultured NK cells was insufficient to account for the observed disappearance of this subset. Our observations in primates are similar to those seen in LCMV-infected mice, where decreased circulating NK cells were attributed to margination and cell death. Thus, the disappearance of these cells during acute hemorrhagic fever in rhesus macaques may be a cytokine-induced lymphopenia common to many virus infections.

Infection with Trypanosoma cruzi TcII and TcI in free-ranging population of lion tamarins (Leontopithecus spp): an 11-year follow-up

Here, we present a review of the dataset resulting from the 11-years follow-up of Trypanosoma cruziinfection in free-ranging populations of Leontopithecus rosalia(golden lion tamarin) andLeontopithecus chrysomelas(golden-headed lion tamarin) from distinct forest fragments in Atlantic Coastal Rainforest. Additionally, we present new data regarding T. cruziinfection of small mammals (rodents and marsupials) that live in the same areas as golden lion tamarins and characterisation at discrete typing unit (DTU) level of 77 of these isolates. DTU TcII was found to exclusively infect primates, while TcI infectedDidelphis aurita and lion tamarins. The majority ofT. cruziisolates derived from L. rosaliawere shown to be TcII (33 out 42) Nine T. cruziisolates displayed a TcI profile. Golden-headed lion tamarins demonstrated to be excellent reservoirs of TcII, as 24 of 26 T. cruziisolates exhibited the TcII profile. We concluded the following: (i) the transmission cycle of T. cruziin a same host species and forest fragment is modified over time, (ii) the infectivity competence of the golden lion tamarin population fluctuates in waves that peak every other year and (iii) both golden and golden-headed lion tamarins are able to maintain long-lasting infections by TcII and TcI.

Inquérito sorológico para toxoplasmose e leptospirose em mamíferos selvagens neotropicais do Zoológico de Aracaju, Sergipe

Os zoológicos modernos são instituições destinadas à manutenção da fauna selvagem com o objetivo de promover a conservação, pesquisa científica, lazer, recreação e educação ambiental. A ampla variedade de espécies selvagens, vivendo em condições diferentes do seu habitat natural, representa um ambiente propício à disseminação de doenças, muitas delas zoonóticas. Devido à escassez de dados e à relevância dos mamíferos selvagens neste contexto epidemiológico, tanto na toxoplasmose, quanto na leptospirose, foi efetuado o inquérito sorológico para toxoplasmose e leptospirose em mamíferos selvagens neotropicais do Zoológico de Aracaju, Sergipe, Brasil. Para tanto foram colhidas amostras sanguíneas de 32 animais, adultos, de ambos os sexos incluindo: 14 macacos-prego (Cebus libidinosus), quatro macacos-prego-do-peito-amarelo (Cebus xanthosternus), três onças-suçuaranas (Puma concolor), uma onça-pintada (Pantheraonca), uma raposa (Cerdocyon thous), seis guaxinins (Procyon cancrivorus), dois quatis (Nasua nasua) e um papa-mel (Eira barbara). Para a pesquisa de anticorpos anti-Toxoplasma gondii foi utilizado o Teste de Aglutinação Modificada (MAT ³"1:25) e para pesquisa de anticorpos anti-Leptospira spp. foi utilizado o teste de Soroaglutinação Microscópica (ponto de corte ³1:100) com uma coleção de antígenos vivos que incluiu 24 variantes sorológicas de leptospiras patogênicas e duas leptospiras saprófitas. Dentre os 32 mamíferos, 17 (53,1%) apresentaram anticorpos anti-T. gondii e quatro (12,5%) foram positivos para anticorpos anti-Leptospira spp. De acordo com o sexo, 60% (9/15) dos machos e 47,1% (8/17) das fêmeas foram soropositivos para T. gondii e 26,7% (4/15) dos machos apresentaram anticorpos anti-Leptospira spp. Dos mamíferos que apresentaram anticorpos anti-T. gondii, 47% (8/17) nasceram no zoológico, 41,2% (7/17) foram oriundos de outras instituições e dois (11,8%) foram provenientes da natureza. Em relação aos quatro mamíferos soropositivos para Leptospira spp., três (75%) foram procedentes da natureza e um (25%) nasceu no zoológico. Este foi o primeiro inquérito sorológico de anticorpos anti-Leptospira spp. em primatas e carnívoros neotropicais em um zoológico do Nordeste do Brasil e descreveu pela primeira vez a ocorrência de anticorpos anti-T. gondii e anti-Leptospira spp. com sorovar mais provável Copenhageni no primata ameaçado de extinção macaco-prego-de-peito-amarelo (C. xanthosternus) em Aracaju, SE.

Comparative anatomy of the thigh nerves of Cebus libidinosus (Rylands et al., 2000)

Non-human primates have constituted an important group among animals subjected to various studies. Ethological, evolutionary and paleontological studies have revealed changes in anatomical structures linked to the evolution of primates, considered in studies on the comparative anatomy between Cebus libidinosus and other neotropical monkeys or those from the Old World, and the detailed knowledge on their anatomy may represent an important factor for their preservation and protection when the animals are brought to veterinary clinics after accidents or illnesses. In terms of veterinary importance, sometimes these animals arrive in the veterinary medical clinics after accidents, needing surgery or clinical treatment, but the little data available on anatomy has impaired the correct proceedings. The main justification for studies on C. libidinosus, is due to little information about the anatomy related to C. libidinosus in Brazilian and worldwide scientific literature. In this study, the distribution, enervation and path of the femoral and sciatic nerves of the pelvic limb (thigh) of C. libidinosus were studied and these results were compared with literature on the anatomy of humans, chimpanzees and baboons. In general, the enervation model of the four primates is identical, but in specific terms, the differences in enervations indicate evolution convergence closer to the branch of baboons in the evolutionary tree, and these data corroborate other comparative studies in relation to the same primates to vessels, muscles and nerves. In conclusion, the nerve organization in the thigh of C. libidinosus is identical to baboon, chimpanzee and homo, but more similar to baboon. The specific differences found indicate an ancient phylogenic origin to C. libidinosus and baboons (data corroborated by other studies).

Salmonella Yoruba infection in white-tufted-ear marmoset (Callithrix jacchus)

The aim of this study was to describe a fatal salmonellosis case in a non-human female primate (Callithrix jacchus), found in the illegal pet trade in Brazil. The marmoset was sent to the quarantine section of the Guarulhos City Zoo and died in the sequence of an episode of profuse diarrhea. Necropsy findings included mucous enteritis, and liver enlargement and necrosis. Feces and liver fragments were collected for bacteriological tests, which indicated the presence of Salmonella sp.; it was subsequently characterized as pertaining to the Yoruba serotype. The susceptibility profile demonstrated resistance to tetracycline only. The strain was positive for genes that encoded the virulence factors investigated (invA, sefC, pefA and spvC). The results indicated the risk of introduction of Salmonella pathogenic serotypes in primates in captivity.

Macroscopic aspects of Saimiri sciureus dura mater

Saimiri sciureus is a small New World primate (NHP) commonly called macaco-de-cheiro that inhabits the tropical forests of the Amazon basin. Anatomical features are not well studied in most primates, and the encephalic morphology and related structures are still quite unknown. Comparative anatomy of the meninges in South American primates is still scarce. Dura mater, arachnoid and pia mater are a group of stratified layers that surrounds and promotes protection to the medulla spinalis. This study aimed to shed light on the anatomy of dura mater in Samiri sciureus in order to contribute to the neuroscience in primates. We investigated three young females and two males of S. sciureus. Specimens were fixed through perfusion with a 10% formaldehyde aqueous solution. In S. sciureus encephalus few gyrus and circunvolutions, and a very delicate system consisting of eight sinus venosus was found between the dura mater layers. Based on our findings, we can conclude that the Saimiri sciureus dura mater is quite similar to other mammals, however we detected a new sinus venosus formation at the level of parietal bone, named sinus parietalis, what appears to be its first description.

Saimiri sciureus' hard palate morphology

Primate order includes around 180 species. Morphological aspects of New World non-human primates (NHP) have been extensively investigated since last century. General commonsense describes oral cavity adaptations according to diet and feeding, dentition, tongue projection and head shape. Morphological appearance and dimension of the hard palate have been outstanding as interest in many species including man. Six young Saimiri sciureus hard palate were investigated. We measured the hard palate distance (HL), intercanine distance (ICD), intermolar distance (IMD), and interpremolar distance (IPD). Complete and incomplete palatine crests were quantified. We believe that better understanding of the mouth roof morphology will contribute to improve the management of captive animal's diet in order to re-introduce the animals in its habitat.

Epidemiology of leptospirosis at Sorocaba Zoo, São Paulo state, Southeastern Brazil

Leptospirosis is considered a worldwide distributed zoonosis, caused by the bacteria Leptospira spp. Since several species of wildlife animals are reportedly reservoirs, the aim of the present study was to know the epidemiology of leptospirosis at the Sorocaba Zoo, Southern Brazil. Serum samples of wild mammals from Artiodactyla, Carnivora, Didelphimorphia, Diprotodontia, Perissodactyla, Pilosa, Primates, Proboscidea and Rodentia orders, kept in captivity as well as from zoological staff were assayed by microscopic agglutination test (MAT). Whole blood, urine and tissue samples from wild mammals and synanthropic animals were assayed by polymerase chain reaction (PCR). An epidemiological survey was applied to evaluate the risk factors for animal infection and staff level of knowledge on leptospirosis. A total of 13/229 (5.68%; CI95% 3.37-9.47%) serum samples from wild mammals were reagent on MAT. Serology from synanthropic animals, zoo staff and molecular analysis of animal samples were all negative. Leptospirosis knowledge of zoo park staff was considered medium. In conclusion, leptospiral infection occurs at the studied zoo but due to the low occurrence found, the lowest reported in literature, wild captive mammals do not act as source of infection of leptospirosis to other animals and human beings.

Comparative anatomical study of the leg's nerves of Cebus (barbed capuchins) with baboons, chimpanzees and modern humans

The anatomical comparative studies among the primates are important for the investigation of ethology, evolution, taxonomy, and comprehension of tools by hominoids. Especially the anatomical knowledge of Cebus contributes to conservation of the species, and to development of surgical procedures and clinical treatments of these animals, as they frequently are victims of automobile accidents. Recent anatomical studies came to a wrong conclusion regarding behavioral traits of Cebus, ascribed to few data available in previous literature. Therefore, to provide anatomical data and to support the other sciences related to anatomy, and to develop surgical and/or clinical procedures, we described the nerves of the legs of Cebus foccusing on their position and trajectory, as wll as innerved muscles, and compared these results with those of humans and other primates. Eight adult capuchin specimens were used for this study. The anatomical comparative study of the leg's nerves of Cebus demonstrated that, in general, structural organization of the nerves is similar among the four primates analyzed here (Cebus, chimpanzees, baboons and humans), which might be attributed to the fact that the all four primates have similar body structures. However, nerve trajectory and muscles innervation in Cebus was more similar to baboons.

Frequency of Toxoplasma gondii antibodies in tufted capuchin monkeys (Cebus apella nigritus) from an ecological station in the State of São Paulo, Brazil

Toxoplasmosis is a worldwide zoonosis caused by Toxoplasma gondii, an obligate intracellular parasite protozoan. A large percentage of animals presents specific antibodies caused by a previous exposition, resulting in a chronic infection. Felides are the definitive hosts and the other warm-blooded animals, including primates, are the intermediate hosts. This study was aimed to determine the prevalence of T. gondii infection in free-living tufted capuchin monkeys (Cebus apella nigritus) from an ecological station located on Mata de Santa Teresa, Ribeirão Preto, SP, Brazil. T. gondii antibodies were analyzed by modified agglutination test (MAT) in serum samples of 36 tufted capuchin monkeys, considering eight as cut-off titer. From the studied animals, 3/36 (8.33%; CI95% 3.0-21.9%) presented T. gondii antibodies, all with titer 32. No significative difference was observed relating to the sex (1/3 male and 2/3 female), and to the age (1/3 young and 2/3 adult) (P>0.05). Thus, these results demonstrate the presence of T. gondii antibodies in primates from São Paulo state.

Diagnosis of Leishmania infantum infection by Polymerase Chain Reaction in wild mammals

Visceral leishmaniasis is a chronic infectious disease caused by Leishmania infantum (synonym: Leishmania chagasi) and transmitted by the sandfly Lutzomyia longipalpis in Brazil. It is an endemic zoonosis in several regions of the country, including Belo Horizonte (State of Minas Gerais). In urban areas, the domestic dog is susceptible and considered the most important animal reservoir. However, L. infantum has been previously diagnosed in other species, including captive primates and canids. This study aimed to evaluate the presence of the agent DNA in captive animals as well as some free ranging animals from the Zoo-Botanical Foundation of Belo Horizonte by Polymerase Chain Reaction. Eighty one blood samples from primates, carnivores, ruminants, edentates, marsupial, and a monogastric herbivore were analyzed. Three primates Alouatta guariba (brown howler monkey), and two canids Speothos venaticus (bush dog) were positive, demonstrating the importance of leishmaniasis control in endemic areas for preservation of wildlife species in captivity.

Relation between the level of self-mutilation and theconcentration of fecal metabolites of glucocorticoids incaptive chimpanzees (Pan troglodytes)

The influence of stress in an environment, according with the behavioral and endocrine variables of primates, are increasingly being studied by a diversity of authors, and have shown that abnormal behaviors associated with increased glucocorticoids may be directly related with the impairment of their well-being. In this work were used 22 adult chimpanzees (Pan troglodytes), 11 males and 11 females, kept in captivity in three different institutions. All animals had their behavior registered by focal session using a 30 seconds sample interval, during six months, totaling 4,800 registries per each animal. During this period, fecal samples were collected 3 times a week for the extraction and measurement of the concentration of fecal metabolites of glucocorticoid by radioimmunoassay. Of the total observed, stereotypical behaviors represented 13,45±2.76%, and among them, self-mutilation represented 38.28±3.98 %. The animals were classified into three different scores, according with the percentage of body surface with alopecia due to self-mutilation. It was found a positive correlation of high intensity between the scores of alopecia due to the observed mutilation and the average concentrations of fecal metabolites of glucocorticoids. This result strongly suggests that this measurement of self-mutilation in a chimpanzee can be used as an important auxiliary tool to evaluate de conditions of adaptation of an animal in captivity, functioning as a direct indicator of the presence of chronic stress.

Estrutura genética intrapopulacional em Cryptocarya moschata Nees (Lauraceae)

Pela análise de seis sistemas enzimáticos (ACP, ALP, CAT, GOT, PPO e PO), com 18 locos polimórficos, foram calculadas as freqüências alélicas de 24 alelos referentes a 141 indivíduos adultos de uma população natural de Cryptocarya moschata (noz-moscada-do-Brasil) do Parque Estadual Carlos Botelho (24º03'21" S, 47º59'36" W), São Miguel Arcanjo, SP, amostrados em uma área de cerca de 647 ha. A estrutura genética espacial foi investigada através do método de autocorrelação espacial, utilizando-se coeficientes I de Moran estimados para 10 classes de distância. Adicionalmente, para as mesmas classes de distância, estimaram-se os coeficientes r através de uma análise multivariada. Ambas as abordagens mostraram-se similares qualitativamente, sendo que 15 correlogramas de coeficientes I foram significativos ao nível de 95% de probabilidade. O correlograma total gerado pelos coeficientes r indicou estruturação espacial significativa para a classe de distância de 0 a 150 m. Pela subdivisão desta classe em intervalos de 10 m, detectou-se autocorrelação espacial positiva na maioria das classes, indicando que os agrupamentos de árvores de C. moschata, que se encontram separadas a distâncias menores que 50 m, são constituídos por indivíduos semelhantes geneticamente e provavelmente aparentados. Contudo, a maior similaridade genética ocorreu entre indivíduos distanciados em torno de 105 m, podendo estar associada à dispersão de sementes promovida por Brachyteles arachnoides (Primates - Cebidae). Pela análise dos resultados, conclui-se que para uma amostragem adequada, visando a detecção de diversidade genética, a coleta de indivíduos distanciados acima de 750 m seria recomendável.