Pesquisa de genomas virais em primatas não humanos do novo mundo por metagenômica

Pós-graduação em Medicina Veterinária - FMVZ

Estudo experimental sobre a resposta imunológica em infecções seqüenciais pelo vírus Dengue 3 e pelo vírus Dengue 2 em primatas não humanos da espécie Callithrix penicillata

A dengue é causada pelo Vírus da dengue (VDEN - Flaviviridae, Flavivirus) e é considerada a arbovirose mais difundida no mundo. Atualmente, não há um animal que sirva como modelo para estudos sobre a patogênese do VDEN. Para investigar a susceptibilidade da espécie Callithrix penicillata ao VDEN foram inoculados amostras do VDEN-3 e do VDEN-2 de isoladas de casos humanos fatais. Para tal, vinte e dois animais foram infectados com VDEN-3 (3,23 × 103 PFU/mL) e, sessenta dias após a infecção (d.p.i.), 11 deles foram secundariamente infectados com VDEN-2 (4,47 × 104 PFU/mL). Sangue, plasma e soro, foram coletados diariamente durante os primeiros sete dias e em 15, 20, 45 e 60 d.p.i.. Foram investigados a produção de anticorpos IgM e inibidores da hemaglutinação, assim como foram análisados parâmetros hematologicos e bioquímicos e o perfil sérico de citocinas (IL-6, TNF-, IL-2, IFN-α, IL-4 e IL-5). A infecção primária (VDEN-3) revelou anticorpos IgM (15- 20 d.p.i.), anticorpos IH (15-60 d.p.i.), aumento dos níveis de TNF-α e IFN-γ e diminuição da IL-5. Na infecção secundária (VDEN-2), foi detectado anticorpos IgM (15-20 d.p.i.), anticorpos IH (15-60 d.p.i.) e diminuição dos níveis de IL-6, TNF-α, de IFN-γ e IL-5. Além disso, foi observado em ambas infecções leucopenia, neutropenia, linfocitopenia, monocitopenia, trombocitopenia, e aumentou da AST. O aumento dos níveis de INF-γ e TNF- α indicaram a ativação da resposta inflamatória, com a diferenciação para a resposta celular e supressão da proliferação de citocinas características da resposta humoral. A presença de anticorpos neutralizantes pode ter ocasionado a supressão da resposta inflamatória na infecção secundária o que pode ter levado a ausência de sinais de Febre Hemorrágica do Dengue/Sindrome do Choque do Dengue (FHD/SCD) durante a infecção secundária (VDEN- 2). Os resultados indicam que o primatas não humanos da espécie Callithrix penicillata demonstraram susceptibilidade à cepas de VDEN de origem humana, sugerindo que esses animais são um bom modelo para estudos da resposta imunológica da Febre do Dengue (FD), assim como para a avaliação de uma vacina tetravalente.

Modelo experimental de imunossupressão com ciclofosfamida em Rattus norvegicus da linhagem wistar e primatas não humanos da especie Cebus apella: análise genotoxicológica

Foi estabelecido um modelo de imunossupressão em roedores por inoculação do agente alquilante Ciclofosfamida (CY). A administração de 50 mg/kg de CY em ratos Wistar provocou uma significante diminuição dos parâmetros de celularidade e peso relativo dos órgãos linfóides. Pela análise da titulação de anticorpos, do ensaio sobre as células formadoras de placa e do teste de hemólise foi comprovada que a imunidade humoral dos roedores sofreu supressão. Foram realizadas quatro inoculações desse imunossupressor e a periodicidade entre as inoculações foi determinada pela recuperação dos níveis de normalidade dos parâmetros supracitados. A alteração na contagem diferencial de células sanguíneas brancas representou o maior efeito adverso da CY, observado nos parâmetros de laboratório analisados nos Cebus apella. Nas duas vezes que foi administrada a droga houve redução no número de linfócitos e posteriormente diminuição de neutrófilos, porém somente na segunda foi observada a imunossupressão. Visto a proximidade filogenética dos primatas não humanos, este desenho experimental será de suma importância para o estudo de tumores em diversas fases do desenvolvimento e principalmente para testes de novos fármacos e esquemas terapêuticos. Com relação às análises de genotoxicidade da CY podemos concluir que em ratos Wistar, as administrações de CY aumentaram significativamente a freqüência de micronúcleos em eritrócitos policromáticos (MN PCEs) e provocaram efeito citotóxico (P<0.05). Em C. apella, os linfócitos do sangue periférico, após o tratamento com CY apresentaram um aumento significativo da media de MN/1000 células em relação aos linfócitos controle (P<0.05). A concentração de CY de 50mg/kg, em C. apella, corresponde à concentração DL50 da droga, visto que 50% desses animais morreram durante o experimento de imunossupressão. Até o desenvolvimento deste trabalho, não se conhecia a concentração correspondente ao DL50 nessa espécie. Ao comparamos as duas espécies de animais utilizadas neste trabalho, os primatas não humanos têm uma recuperação imune mais rápida em relação aos ratos Wistar. Provavelmente a capacidade de metabolização da droga seja mais eficaz em C. apella. Nossos resultados apóiam, portanto, que os primatas não humanos constituem os melhores modelos experimentais devido a sua grande proximidade evolutiva e filogenética com o ser humano.

Estudo da distribuição das proteínas relacionadas às teneurinas no sistema nervoso central de primatas não-humanos (Sapajus spp) e ratos (Rattus norvegicus)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Estudo da distribuição das proteínas relacionadas às teneurinas no sistema nervoso central de primatas não-humanos (Sapajus spp) e ratos (Rattus norvegicus)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Diagnóstico morfológico, sorológico e molecular de Plasmodium spp. em primatas neotropicais na Ilha de São Luís, Estado do Maranhão, Brasil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Estudo comportamental, hormonal e histopatológico de animais com perda de peso progressina (SEP) e da variável peso associada a uma nova classificação ontogenética em Callithrix jacchus

The use of non-human primates in scientific research has contributed significantly to the biomedical area and, in the case of Callithrix jacchus, has provided important evidence on physiological mechanisms that help explain its biology, making the species a valuable experimental model in different pathologies. However, raising non-human primates in captivity for long periods of time is accompanied by behavioral disorders and chronic diseases, as well as progressive weight loss in most of the animals. The Primatology Center of the Universidade Federal do Rio Grande do Norte (UFRN) has housed a colony of C. jacchus for nearly 30 years and during this period these animals have been weighed systematically to detect possible alterations in their clinical conditions. This procedure has generated a volume of data on the weight of animals at different age ranges. These data are of great importance in the study of this variable from different perspectives. Accordingly, this paper presents three studies using weight data collected over 15 years (1985-2000) as a way of verifying the health status and development of the animals. The first study produced the first article, which describes the histopathological findings of animals with probable diagnosis of permanent wasting marmoset syndrome (WMS). All the animals were carriers of trematode parasites (Platynosomum spp) and had obstruction in the hepatobiliary system; it is suggested that this agent is one of the etiological factors of the syndrome. In the second article, the analysis focused on comparing environmental profile and cortisol levels between the animals with normal weight curve evolution and those with WMS. We observed a marked decrease in locomotion, increased use of lower cage extracts and hypocortisolemia. The latter is likely associated to an adaptation of the mechanisms that make up the hypothalamus-hypophysis-adrenal axis, as observed in other mammals under conditions of chronic malnutrition. Finally, in the third study, the animals with weight alterations were excluded from the sample and, using computational tools (K-means and SOM) in a non-supervised way, we suggest found new ontogenetic development classes for C. jacchus. These were redimensioned from five to eight classes: infant I, infant II, infant III, juvenile I, juvenile II, sub-adult, young adult and elderly adult, in order to provide a more suitable classification for more detailed studies that require better control over the animal development

Saguis (Callithrix jacchus) sob ciclo claro-escuro de 21 h: um modelo de dessincronização forçada em primata diurno

The circadian system consists of multiple oscillators organized hierarchically, with the suprachiasmatic nucleus (SCN) as the master oscillator to mammalians. There are lots of evidences that each SCN cell is an oscillator and that entrainment depends upon coupling degree between them. Knowledge of the mechanism of coupling between the SCN cells is essential for understanding entrainment and expression of circadian rhythms, and thus promote the development of new treatments for circadian rhythmicity disorders, which may cause various diseases. Some authors suggest that the dissociation model of circadian rhythm activity of rats under T22, period near the limit of synchronization, is a good model to induce internal desynchronization, and in this way, enhance knowledge about the coupling mechanism. So, in order to evaluate the pattern of the motor activity circadian rhythm of marmosets, Callithrix jacchus, in light-dark cycles at the lower limit of entrainment, two experiments were conducted: 1) 6 adult females were submitted to the LD symmetric cycles T21, T22 and T21.5 for 60, 35 and 48 days, respectively; 2) 4 male and 4 female adults were subjected to T21 for 24 days followed by 18 days of LL, and then back to T21 for 24 days followed by 14 days of LL. Vocalizations of all animals and motor activity of each one of them were continuously recorded throughout the experiments, but the vocalizations were recorded only in Experiment 1. Under the Ts shorter than 24 h, two simultaneous circadian components appeared in motor activity, one with the same period of LD cycle, named light-entrained component, and the other in free-running, named non-light-entrained component. Both components were displayed for all the animals in T21, five animals (83.3%) in T21.5 and two animals (33.3%) in T22. For vocalizations both components were observed under the three Ts. Due to the different characteristics of these components we suggest that dissociation is result of partial synchronization to the LD cycle, wherein at least one group oscillator is synchronized to the LD by relative coordination and masking processes, while at least another group of oscillators is in free-running, but also under the influence of masking by the LD. As the T21 h was the only cycle able to promote the emergence of both circadian components in circadian rhythms of all Callithrix jacchus, this was then considered the lower entrainment limit of LD cycle promoter of dissociation in circadian rhythmicity of this species, and then suggested as a non-human primate model for forced desynchronization

Antimicrobial effect of human serum on oral Fusobacterium nucleatum isolates from humans and monkeys

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Language evolution and recursion : an empirical investigation of human hierarchical processing

Tese de doutoramento, Ciências Biomédicas (Neurociências), Universidade de Lisboa, Faculdade de Medicina, 2014

Início da função gonadal no sagui comum (Callithrix jacchus): diferenças entre gêneros

Os primatas não humanos, como Callithrix jacchus vêm sendo utilizados com maior frequência em pesquisa biomédica e, recentemente, nosso laboratório propôs uma nova classificação para monitorar com mais detalhes as modificações comportamentais e hormonais relacionadas ao desenvolvimento. O presente trabalho monitorou os níveis de hormônios gonadais de 4 machos e 6 fêmeas de C. jacchus durante as fases juvenil I e II e subadulta (dos 5 aos 15 meses). Os resultados apontaram que os níveis de progesterona nas fêmeas e andrógenas nos machos foram estatisticamente mais elevados nos estágios juvenil II e subadulto, respectivamente, em relação ao estágio juvenil I. Estes achados indicam que, de acordo com a nova classificação, foi possível observar que o funcionamento gonadal se instala mais cedo nas fêmeas

Estudo da preferência manual durante o desenvolvimento ontogenético em sagüi comum ( Callithix jacchus)

The study of manual preference is a widely used approach to investigate cerebral laterality in nonhuman primates. However, in New World primates, little is known about the ontogenesis of hand use asymmetry, in both forced and spontaneous activities, as well as how they correlate with sexual hormones. Accordingly, a longitudinal study was conducted on the manual preference of 6 female and 4 male common marmosets (Callithrix jacchus). The study included the record of forced tests to reach for the food using only one hand (forced activity) and activities such as grooming (auto and social), scratching, grasping the food and hanging, in weekly sessions from the infantile (4 months) to the early adult phase (15 months). Feces samples were also collected, at least once a week, to evaluate the level of gonadal steroids and their influence on these behaviors. In the forced activity, the results confirm the influence of the development period on manual preference during feeding, shown by the increase in lateral stability when grasping the food between the juvenile and adult phases. During this period, a sexual hormone effect on development was also observed, mainly of progesterone in females and androgen in males, but no difference between sexes was found. In the females, progesterone also influenced the manual preference index, with a proportional increase in the degree of manual asymmetry during puberty. With respect to spontaneous activities, the animals showed proportional use of both hands when scratching, hanging, holding the food and grooming. A positive correlation was also found between the preference for holding the food in forced activities and in spontaneous activities

A influência da recompensa e da experiência na discriminação visual simultânea em sagüi comum (Callithrix jacchus)

The discrimination learning is assessed through instrumental tasks in which the individual is rewarded for choosing one item over another. Thus, in concurrent visual discrimination of objects the animal must learn that only one of the objects will be rewarded. The concurrent visual discrimination is relatively simple, and already been observed Callithrix jacchus is able to accomplish this task. As yet wasn't seen the influence of the qualitative aspects of the rewards, in the performance of concurrent visual discrimination of objects in nonhuman primates, and as in most tests are used isolated animals, the present study had two stages: at first we had as objective to analyze the influence of the caloric value of the reward on the performance in concurrent visual discrimination of objects in isolated animals; in the second, we had the intention analyze performance of C. jacchus in realization of discrimination task in different social contexts, as well as, analyze the influence of previous experience in task performance. In the first stage (Study 1), the animals were not able to discriminate foods that presented small caloric differences . This incapacity in discriminates the rewards was responsible by generating randomness in task of concurrent visual discrimination of objects. In the second stage (Study 2), observed that, independent of social context in which the task was presented, the performance both of the experienced animals as the inexperienced animals tended to randomness. In the first case, is likely that the pattern of responses of the experienced animals is a reflection of their own performance when they were observed in isolation. In the second case, in turn, the randomness was probably due to the small number of sessions. Although present a pattern of performance similar to inexperienced individuals, we verify that the experienced animals monopolize the food consumption when they were in the presence of inexperienced individuals. This was a consequence of the experienced animals have presented lower latency the approximation of apparatus and, consequently, obtain more food. In turn, the inexperienced animals, when were in the presence of experienced, had to adopt alternative strategies to obtain food. Thus, C. jacchus is able to use the previous information he had about the task of solving their own benefit.

O papel das relações sociais e da atenção social durante tarefas cooperativas em Callithrix jacchus

Several studies on nonhuman primates show that the relationships between individuals strongly influence the expression of cooperative behavior, both in natural environment and in captivity settings. Recent studies suggest that cooperative breeders present outstanding performance in tasks involving social cognition, such as cooperative tasks with experimental apparatuses. In experimental research on this subject it is crucial to differentiate between real cooperation (or communicative cooperation, mediated by social attention) and by-product cooperation that results from simultaneous actions of individuals. The present study assessed, in Callithrix jacchus, a cooperative breeder species, if social relationships and social attention between subjects are important factors during performance in cooperative tasks. During the experimental procedure the animals participated in three different cooperative tasks: cooperation task, prosocial task and control task. Diverging from the literature, matrix correlation tests revealed no significant relationship between grooming or proximity and the execution of the tasks, suggesting that other factors such as age or hierarchy may have an effect on the performance in cooperative tasks in this species. There was also no relationship between the execution of the cooperative tasks and social glances, suggesting that there was no social attention during the tasks. Moreover, there were lower rates of social glances in the cooperative tasks as opposed to the control tasks. However, the small number of pulls in prosocial tasks suggests that the animals distinguished between tasks that benefited only a partner and tasks that generated benefits to themselves, choosing the latter. We conclude that, for the tasks presented in this study, we could neither detect the role of social relationships on the cooperative tasks nor assert that there were true cooperation and prosocial behavior

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

A toxoplasmose é uma das zoonoses mais difundidas no mundo, causada pelo Toxoplasma gondii, um protozoário parasita intracelular obrigatório. Uma alta porcentagem de animais apresenta anticorpos específicos causados por exposição prévia, levando a uma infecção crônica. Os felídeos são os hospedeiros definitivos e outros animais homeotérmicos, incluindo os primatas, são os hospedeiros intermediários. Este estudo objetivou determinar a prevalência da infecção por T. gondii em macacos-prego (Cebus apella nigritus) de vida livre da Estação Ecológica localizada na Mata de Santa Teresa, Ribeirão Preto, SP, Brasil. Anticorpos anti-T. gondii foram pesquisados pelo método de aglutinação direta modificada (MAT) em amostras de soro de 36 macacos-prego, utilizando-se o título oito como de corte. Dos animais estudados, 3/36 (8,33%; IC95% 3,0-21,9%) apresentaram anticorpos anti-T. gondii, todos com título 32. Nenhuma diferença significativa (P>0,05) foi observada com relação ao sexo (1/3 machos e 2/3 fêmeas), e à idade (1/3 jovens e 2/3 adultos). Assim, estes resultados demonstram alta prevalência de anticorpos anti-T. gondii em primatas no estado de São Paulo.