Propondo um problema de forrageio como meio de enriquecer o cativeiro: um estudo comparativo de duas espécies de primatas brasileiros (Callithrix penicillata e Saguinus imperator)

O presente estudo analisou a influência de um simples aparelho que simulou o forrageamento ativo por insetos, funcionando como um alimentador enigmático denominado puzzle, sobre o comportamento de duas espécies de calitriquíneos (Callitrichinae, Primates) mantidos em cativeiro no Centro Nacional de Primatas (CENP). Buscou-se comparar a reação das duas espécies frente ao aparelho proposto, e verificar a viabilidade deste artifício no enriquecimento ambiental das condições de cativeiro para estes primatas. Foram utilizados três casais de Saguinus imperator, conhecido por sagui imperador e três casais de Callithrix penicillata, conhecido por mico estrela, as comparações foram feitas entre as espécies e entre os sexos. Foram gravadas através de vídeo cassete seções de observação com duração de três horas, divididas em dois períodos, um controle (ausência do puzzle) e outro experimental (presença do puzzle), com um total de 36 horas de observação para cada casal. As sessões de controle foram usadas para calcular a linha base do orçamento de atividade para comparações com padrões de comportamento durante as sessões experimentais. As fitas foram transcritas, e todos os eventos de comportamento foram cronometrados, e registrados as medidas da freqüência e duração de eventos. Nas duas espécies a manipulação do puzzle não alcançou uma proporção de tempo muito elevada, porém demonstrou uma diferença clara entre as duas espécies, ocupando 3,96% do tempo total dos micos e 1,99% do tempo dos saguis. Os micos gastaram com repouso, durante as seções experimentais, 17% menos tempo em comparação com a situação controle. Considerando que a ociosidade reduziu em menos de 7% nos saguis, com a atividade geral aumentada em 10%. Entre os sexos, notou-se uma inversão, onde as fêmeas de sagui tiveram um aumento de 18% na ociosidade e queda de 14% na atividade, enquanto as fêmeas de mico tiveram uma diminuição nas duas categorias de pouco menos de 10%. Os machos responderam mais ao puzzle, os saguis aumentaram em 58% a atividade e diminuíram em 23% a ociosidade, os micos aumentaram a atividade em 4% e diminuíram a ociosidade em menos de 10%. Todos os animais aprenderam a manipular o equipamento e capturar os insetos, sendo que os micos tiveram um maior êxito no número de larvas capturadas, superando aos saguis em cerca de 54% de seu índice. As fêmeas tiveram mais êxitos em ambas as espécies, porém superficialmente nos micos. Em contraste, nos saguis as fêmeas foram 16% mais prósperas que os machos. O puzzle foi eficiente em enriquecer o ambiente cativo dos animais e estimular o comportamento manipulativo, sendo um instrumento importante na busca do bem estar para as duas espécies.

Diagnóstico populacional e comportamental de saguis-de-tufo-preto (Callithrix penicillata) em ambiente peri-urbano

The black-tufted-marmoset (Callithrix penicillata), abundant in the southeastern state of São Paulo, is an exotic and invasive species . The present study aimed to study the behavior and make monitoring a population black-tufted-marmoset in man-made environment. The observations were made following the method of scan sampling, which consists of continuous observations with a duration of three minutes with a fifteen-minute intervals between one record and another. The individuals are monitored at two locations on campus, where two groups were identified (with a mean of 3.50 ± 2.06 individuals) totaling 291 hours and 30 minutes of observation. Were determined tree layers used, and observed that the group of EC preferably utilizes the highest stratum, while the group of LCF uses the middle and lower strata. Types of vocalization, such as calling, interaction and alert and frequency and percentage of these were also observed. The food items consumed by the groups that are similar in the two study sites, mostly sap and leaves were diagnosed. From observations of behavioral patterns was possible to construct an ethogram of the species ' Luiz de Queiroz ' campus, comprising the behaviors of vocalization, locomotion, foraging, game, among others. The results may be useful for development of management techniques and for subsequent studies

Avaliação da influência do enriquecimento ambiental nas variáveis comportamentais de saguis-de-cara-branca (Callithrix geoffroyi) em cativeiro

Six species of the genus Callithrix can be found across the Atlantic Forest extension, being the Callithrix geoffroyi, Geoffroy`s marmoset, the least studied species. The objective of this study is to investigate the behavioral aspects of the species in captivity, and how the environmental enrichment techniques influence those behaviors. To this end, we have studied three specimens of C. geoffroyi, in captivity at Municipal Zoo of Piracicaba, São Paulo. The enclosure includes a 37m2 with under masonry, concrete platforms with external deep, trunks, a floor of grass and soil, e grid in their other three sides. The sampling method was focal per time interval, with records every 30 seconds in sessions of 30 minutes, totaling 30 hours for each animal, which were distributed in a baseline phase, an environmental enrichment phase and post-enrichment phase (n = 3660 records in each phase). The twelve enrichment techniques were applied: the physical environment (branches for perches and hoses fabrics for bedding and vanishing points), cognitive (radio, mirror, puzzle food) and food (insects, flowers and eggs hidden, wrapped). Initially, to determine the period of greatest activity of marmosets, we investigated the pattern of activity for three days (9:00 am to 17:00 pm) consecutively at intervals of 15 minutes, recording the number of active and inactive individuals. The 495 records signaled greater activity in the morning, then selected period for the completion of this work. The ethogram comprised 16 behaviors. The inactivity was the predominant phase behavior baseline and post-enrichment with a significant decrease (h=8,62, p <0.01) in the enrichment phase, and a significant increase in the post enrichment (h=18,15, p <0.001). The same was seen for the use of substrate (grid and ground vs. Trunk and concrete, h=5,09, p <0.001 and h=3,98, p <0.05)... (Complete abstract click electronic access below)

Reporting social behaviours of mixed-species troops formed by Callithrix jacchus and Callithrix penicillata (Primate, Callitrichidae)

In New World primates, mixed-species troops have been reported. Here, we analysed the performance of affiliative and agonistic behaviours of Callithrix jacchus and Callithrix penicillata living in mixed groups. For this purpose, we recorded the interaction of the individuals from two groups located in Bauru city, in the state of Sao Paulo (Brazil). Our data show that in both groups, affiliative behaviours appeared more frequently than agonistic ones. We concluded that there is cohesion inside the mixed-species troops observed. We suggest that a deeper knowledge about the social behaviour of mixed-species troop species certainly may be useful in projects linked with the management of the impact caused by them.

Fluorescent antibody test, quantitative polymerase chain reaction pattern and clinical aspects of rabies virus strains isolated from main reservoirs in Brazil

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

Descrição morfológica do diafragma do sagui-de-tufo-branco (Callithrix jacchus)

O músculo diafragma, encontrado apenas nos mamíferos, é o principal músculo no processo respiratório, servindo de fronteira entre as cavidades torácica e abdominal. Sua importância também ganha destaque em pesquisas realizadas no âmbito dos enxertos, empregando-se diversos tipos de membranas biológicas para o reparo de defeitos diafragmáticos, os quais podem gerar hérnias diafragmáticas. Apesar de muitos estudos já conduzidos para com os primatas não humanos, especialmente no que tange a espécie do novo mundo Callithrix jacchus (Sagui-de-tufo-branco), oriundo do nordeste brasileiro, as pesquisas envolvendo o uso do diafragma em tal espécie é inexistente. Deste modo objetivou-se caracterizar a morfologia e a biometria do diafragma na espécie Callithrix jacchus de ambos os sexos, analisando possíveis divergências estruturais entre machos e fêmeas. Para tal foram utilizados quatros animais, 2 machos e 2 fêmeas, adultos, que vieram a óbito por causas naturais, provenientes de um criadouro comercial. Após fixação em solução de formaldeído 10% os animais foram devidamente dissecados para fotodocumentação e em seguida o diafragma coletado para efetuação da biometria (comprimento e largura) com o uso de um paquímetro e para o processamento histológico por meio da coloração de hematoxilina-eosina e tricrômio de masson, da porção muscular. As mensurações feitas permitiram concluir que não houve diferenças signifcativas entre machos e femeas. A topografia e a presença de três aberturas (forame da veia cava caudal, hiato aórtico e esofágico) na extensão do diafragma corroboram com descrições na literatura classica para outros mamíferos. A presença de um centro tendíneo em "V" difere do encontrado para animais como o peixe-boi e porquinho-da-india, mas é similar ao encontrado para o gambá-de-orelhas-brancas e rato albino. No que diz respeito aos achados histológicos conclui-se que as fibras musculares estão dispostas de forma organizada, apresentam diâmetro grande e núcleos basais, tendo, portanto, características similares do músculo estriado esquelético tanto nos animais machos como nas fêmeas.

Sperm head morphometry in ejaculates of adult marmosets (Callithrix jacchus): a model for studying sperm subpopulations and among-donor variations

In humans and other mammals, sperm morphology has been considered one of the most important predictive parameters of fertility. The objective was to determine the presence and distribution of sperm head morphometric subpopulations in a nonhuman primate model (Callithrix jacchus), using an objective computer analysis system and principal component analysis (PCA) methods to establish the relationship between the subpopulation distribution observed and among-donor variation. The PCA method revealed a stable number of principal components in all donors studied, that represented more than 85% of the cumulative variance in all cases. After cluster analysis, a variable number (from three to seven) sperm morphometric subpopulations were identified with defined sperm dimensions and shapes. There were differences in the distribution of the sperm morphometric subpopulations (P < 0.001) in all ejaculates among the four donors analyzed. In conclusion, in this study, computerized sperm analysis methods combined with PCA cluster analyses were useful to identify, classify, and characterize various head sperm morphometric subpopulations in nonhuman primates, yielding considerable biological information. In addition, because all individuals were kept in the same conditions, differences in the distribution of these subpopulations were not attributed to external or management factors. Finally, the substantial information derived from subpopulation analyses provided new and relevant biological knowledge which may have a practical use for future studies in human and nonhuman primate ejaculates, including identifying individuals more suitable for assisted reproductive technologies. (c) 2012 Elsevier Inc. All rights reserved.

Genomische Analyse von natürlichen Killerzell-Rezeptorgenen in nicht-humanen Primaten

Natürliche Killerzell-Rezeptoren, die MHC-Klasse-I-Moleküle binden, sind im Leukozyten Rezeptor Komplex (LRC) und im Natürlichen Killer Komplex (NKC) kodiert. Die Bindung klassischer MHC-Klasse-I-Moleküle erfolgt im Menschen durch die im LRC kodierten polymorphen Killerzell-Immunglobulin-ähnlichen Rezeptoren (KIR) und in Nagetieren durch die im NKC kodierten polymorphen C-Typ Lektin-ähnlichen Ly49-Rezeptoren. Die ebenfalls im NKC kodierten C-Typ Lektin-ähnlichen CD94/NKG2-Rezeptoren sowie der NKG2D-Rezeptor sind sowohl im Menschen als auch in Nagetieren konserviert und wenig polymorph. Im Rahmen dieser Arbeit wurde das CD94-Ly49L-Intervall der NKC-Region in einem Neuweltaffen, dem Weißbüschelaffen (Callithrix jacchus), sowie einem Feuchtnasenaffen, dem Grauen Mausmaki (Microcebus murinus), über Screening von BAC-Banken und Sequenzanalyse von BAC-Contigs untersucht. Das CD94-Ly49L-Intervall im Weißbüschelaffen hat eine Länge von 171 kb und weist orthologe Gene zu den humanen NKC-Genen auf. Eine Ausnahme bildet das Gen NKG2CE, welches äquidistant zu den humanen Genen NKG2C und NKG2E ist. NKG2F und Ly49L sind Pseudogene. Expressionsanalysen der NKC-Gene in neun Weißbüschelaffen-Individuen lieferten einen mäßigen Grad an allelischen Polymorphismen. Alternative Spleißprodukte wurden für CD94, NKG2D und NKG2A identifiziert. Für NKG2A wurden verschiedene Transkripte mit potentiell unterschiedlichen Translationsstartpunkten gefunden. Im Grauen Mausmaki beträgt die Länge des CD94-Ly49L-Intervalls 489 kb. CD94 und die NKG2-Gene sind vervielfacht und wesentlich polymorpher als im Menschen und im Weißbüschelaffen. Expressionsanalysen der NKC-Gene wurden im Grauen Mausmaki und einem weiteren madagassischen Lemuren, dem Schwarzweißen Vari (Varecia variegata), durchgeführt und zeigten, dass CD94 und die NKG2-Gene im Vari ebenfalls vervielfacht sind. Die NKG2-Moleküle der Lemuren weisen unterschiedliche Kombinationen an aktivierenden und inhibierenden Signalmotiven auf und üben somit möglicherweise diverse Funktionen aus. Ly49L stellt in den Lemuren einen potentiell funktionellen inhibierenden Rezeptor dar und NKG2D besitzt im Vergleich zum humanen NKG2D-Protein eine verkürzte Zytoplasmaregion. Alternative Spleißprodukte der NKC-Gene existieren auch in den Lemuren. Darüber hinaus wurden mehrere CD94-Gene in einem weiteren Feuchtnasenaffen, dem Potto (Perodicticus potto) und einem Trockennasenaffen, dem Philippinen-Koboldmaki (Tarsius syrichta), nachgewiesen. Ein Alu-Element, welches ausschließlich in Intron 4 der CD94-Sequenzen des Philippinen-Koboldmakis auftritt, deutet darauf hin, dass sich CD94 in der Linie der Koboldmakis und in der Linie der Feuchtnasenaffen unabhängig voneinander vervielfacht hat. Die vervielfachten, polymorphen CD94/NKG2-Rezeptoren der niederen Primaten stellen möglicherweise das funktionelle Äquivalent zu den polymorphen KIR der höheren Primaten und den polymorphen Ly49-Rezeptoren der Nagetiere dar.

Comparative genomics and phylogenetics of the vertebrate CYP3 family

CYP3A verstoffwechselt mehr als 50% aller gegenwärtig in der Therapie eingesetzten Wirkstoffe, die häufig an klinisch relevanten Arzneimitttel-Wechselwirkungen beteiligt sind. Das Verständnis über die Bedeutung und die Regulation von einzelnen CYP3A Genen in der Pharmakologie und Physiologie ist unvollständig. Wir untersuchten die Evolution des CYP3 Genlokus über einen Zeitraum von 450 Millionen Jahre mittels genomischer Sequenzen von 16 Tierarten. Neue CYP3 Unterfamilien (CYP3B, C und D) entstanden über eine beschleunigte Evolution aus CYP3A Vorstufen von Clupeocephala Spezies. Ausgeprägte funktionelle Unterschiede traten zwischen CYP3A in Säugern und Clupeocephala CYP3 auf. Alle amnioten CYP3A Gene entwickelten sich aus zwei CYP3A Urgenen. Aufgrund der Entstehung von Säugern mit Plazenta ging eines von ihnen verloren während das andere eine neue genomische Umgebung infolge einer Translokation erlangte. In Primaten unterzog sich CYP3A mit mehreren Genduplikationen, Deletionen, Pseudogenisierung und Genkonversionen einer raschen evolutionären Veränderung. Die Entwicklung von CYP3A in Schmalnasenaffen (Alte Welt Affen, große Menschenaffen und Menschen) unterschieden sich wesentlich von Neue Welt Primaten (z.B. gewöhnlichen Krallenaffen) und Feuchtnasenaffen (z.B. Galago). Stellvertretend für die CYP3A Protein-codierende Sequenz entdeckten wir zwei frühe Episoden von besonders starker positiver Selektion: (1) auf CYP3A7 in der frühen hominoiden Evolution, welche im fetalen Zeitraum von einer Einschränkung der hepatischen Expression begleitet war, und (2) auf humanes CYP3A4 im Anschluss an die Teilung der Abstammungslinie in Schimpansen und Mensch. In Übereinstimmung mit diesen Befunden beeinflussen drei von vier positiv ausgewählten Aminosäuren, die in früheren biochemischen CYP3A Studien untersucht wurden, die Aktivität und Regioselektivität. Es ist somit naheliegend, dass CYP3A7 und CYP3A4 katalytische Funktionen erworben haben können, die besonders wichtig waren für die Evolution von Hominoiden und Menschen. Die Charakterisierung von CYP3A Promotoren in Primaten zeigte eine Anreicherung von ER6 Elementen in CYP3A Promotoren von Primaten und einen Trend in Richtung Erhöhung der ER6 Enstehung entlang den Abstammungslinien, die zu humanen und Schimpansen CYP3A4 führten. Die steigende Anzahl an ER6 Elementen kann durch die ausgeprägte CYP3A4 Induzierbarkeit und Expressionsvariabilität im Menschen verursacht sein.

Differential expression of the bone and the liver tissue non-specific alkaline phosphatase isoforms in brain tissues

The enzyme tissue non-specific alkaline phosphatase (TNAP) belongs to the ectophosphatase family. It is present in large amounts in bone in which it plays a role in mineralization but little is known about its function in other tissues. Arguments are accumulating for its involvement in the brain, in particular in view of the neurological symptoms accompanying human TNAP deficiencies. We have previously shown, by histochemistry, alkaline phosphatase (AP) activity in monkey brain vessels and parenchyma in which AP exhibits specific patterns. Here, we clearly attribute this activity to TNAP expression rather than to other APs in primates (human and marmoset) and in rodents (rat and mouse). We have not found any brain-specific transcripts but our data demonstrate that neuronal and endothelial cells exclusively express the bone TNAP transcript in all species tested, except in mouse neurons in which liver TNAP transcripts have also been detected. Moreover, we highlight the developmental regulation of TNAP expression; this also acts during neuronal differentiation. Our study should help to characterize the regulation of the expression of this ectophosphatase in various cell types of the central nervous system.

Molecular and evolutionary genetics of the X -linked visual pigment genes in humans and New World monkeys

Normal humans have one red and at least one green visual pigment genes. These genes are tightly linked as tandem repeats on the X chromosome and each of them has six exons. There is only one X-linked visual pigment gene in New World monkeys (NWMs) but the locus has three polymorphic alleles encoding red, yellow and green visual pigments, respectively. The spectral properties of the squirrel monkey and the marmoset (both NWMs) have been studied and partial sequences of the three alleles are available. To study the evolutionary history of these X-linked opsin genes in humans and NWMs, coding and intron sequences of the three squirrel monkey alleles and the three marmoset alleles were amplified by PCR followed by subcloning and sequencing. Introns 2 and 4 of the human red and green pigment genes were also sequenced. The results obtained are as follows: (1) The sequences of introns 2 and 4 of the human red and green opsin genes are significantly more similar between the two genes than are coding sequences, contrary to the usual situation where coding regions are better conserved in evolution than are introns. The high similarities in the two introns are probably due to recent gene conversion events during evolution of the human lineage. (2) Phylogenetic analysis of both intron and exon sequences indicates that the phylogenetic tree of the available primate opsin genes is the same as the species tree. The two human genes were derived from a gene duplication event after the divergence of the human and NWM lineages. The three alleles in each of the two NWM species diverged after the split of the two NWMs but have persisted in the population for at least 5 million years. (3) Allelic gene conversion might have occurred between the three squirrel monkey alleles. (4) A model of additive effect of hydroxyl-bearing amino acids on spectral tuning is proposed by treating some unknown variables as groups. Under the assumption that some residues have no effect, it is found that at least five amino acid residues, at positions 178 (3 nm), 180 (5 nm), 230 ($-$4 nm), 277 (9 nm) and 285 (13 nm), have linear spectral tuning effects. (5) Adaptive evolution of the opsin genes to different spectral peaks was observed at four residues that are important for spectral tuning. ^

Origins and antiquity of X-linked triallelic color vision systems in New World monkeys

It is known that the squirrel monkey, marmoset, and other related New World (NW) monkeys possess three high-frequency alleles at the single X-linked photopigment locus, and that the spectral sensitivity peaks of these alleles are within those delimited by the human red and green pigment genes. The three alleles in the squirrel monkey and marmoset have been sequenced previously. In this study, the three alleles were found and sequenced in the saki monkey, capuchin, and tamarin. Although the capuchin and tamarin belong to the same family as the squirrel monkey and marmoset, the saki monkey belongs to a different family and is one of the species that is most divergent from the squirrel monkey and marmoset, suggesting the presence of the triallelic system in many NW monkeys. The nucleotide sequences of these alleles from the five species studied indicate that gene conversion occurs frequently and has partially or completely homogenized intronic and exonic regions of the alleles in each species, making it appear that a triallelic system arose independently in each of the five species studied. Nevertheless, a detailed analysis suggests that the triallelic system arose only once in the NW monkey lineage, from a middle wavelength (green) opsin gene, and that the amino acid differences at functionally critical sites among alleles have been maintained by natural selection in NW monkeys for >20 million years. Moreover, the two X-linked opsin genes of howler monkeys (a NW monkey genus) were evidently derived from the incorporation of a middle (green) and a long wavelength (red) allele into one chromosome; these two genes together with the (autosomal) blue opsin gene would immediately enable even a male monkey to have trichromatic vision.

Prevention of autoimmune demyelination in non-human primates by a cAMP-specific phosphodiesterase inhibitor.

Experimental allergic encephalomyelitis (EAE) is an autoimmune disease of the central nervous system that serves as a model for the human disease multiple sclerosis. We evaluated rolipram, a type IV phosphodiesterase inhibitor, for its efficacy in preventing EAE in the common marmoset Callithrix jacchus. In a blinded experimental design, clinical signs of EAE developed within 17 days of immunization with human white matter in two placebo-treated animals but in none of three monkeys that received rolipram (10 mg/kg s.c. every other day) beginning 1 week after immunization. In controls, signs of EAE were associated with development of cerebrospinal fluid pleocytosis and cerebral MRI abnormalities. In the treatment group, there was sustained protection from clinical EAE, transient cerebrospinal fluid pleocytosis in only one of three animals, no MRI abnormality, and marked reduction in histopathologic findings. Rolipram-treated and control animals equally developed circulating antibodies to myelin basic protein. Thus, inhibition of type IV phosphodiesterase, initiated after sensitization to central nervous system antigens, protected against autoimmune demyelinating disease.

First- and second-order stimulus length selectivity in New World monkey striate cortex

Motion is a powerful cue for figure-ground segregation, allowing the recognition of shapes even if the luminance and texture characteristics of the stimulus and background are matched. In order to investigate the neural processes underlying early stages of the cue-invariant processing of form, we compared the responses of neurons in the striate cortex (V1) of anaesthetized marmosets to two types of moving stimuli: bars defined by differences in luminance, and bars defined solely by the coherent motion of random patterns that matched the texture and temporal modulation of the background. A population of form-cue-invariant (FCI) neurons was identified, which demonstrated similar tuning to the length of contours defined by first- and second-order cues. FCI neurons were relatively common in the supragranular layers (where they corresponded to 28% of the recorded units), but were absent from layer 4. Most had complex receptive fields, which were significantly larger than those of other V1 neurons. The majority of FCI neurons demonstrated end-inhibition in response to long first- and second-order bars, and were strongly direction selective, Thus, even at the level of V1 there are cells whose variations in response level appear to be determined by the shape and motion of the entire second-order object, rather than by its parts (i.e. the individual textural components). These results are compatible with the existence of an output channel from V1 to the ventral stream of extrastriate areas, which already encodes the basic building blocks of the image in an invariant manner.

Pyramidal neurons of granular prefrontal cortex of the galago: Complexity in evolution of the psychic cell in primates

Typically, cognitive abilities of humans have been attributed to their greatly expanded cortical mantle, granular prefrontal cortex (gPFC) in particular. Recently we have demonstrated systematic differences in microstructure of gPFC in different species. Specifically, pyramidal cells in adult human gPFC are considerably more spinous than those in the gPFC of the macaque monkey, which are more spinous than those in the gPFC of marmoset and owl monkeys. As most cortical dendritic spines receive at least one excitatory input, pyramidal cells in these different species putatively receive different numbers of inputs. These differences in the gPFC pyramidal cell phenotype may be of fundamental importance in determining the functional characteristics of prefrontal circuitry and hence the cognitive styles of the different species. However, it remains unknown as to why the gPFC pyramidal cell phenotype differs between species. Differences could be attributed to, among other things, brain size, relative size of gPFC, or the lineage to which the species belong. Here we investigated pyramidal cells in the dorsolateral gPFC of the prosimian galago to extend the basis for comparison. We found these cells to be less spinous than those in human, macaque, and marmoset. (c) 2005 Wiley-Liss, Inc.