Induction of Anxiety-Like Phenotypes by Knockdown of Cannabinoid Type-1 Receptors in the Amygdala of Marmosets / 神经科学通报·英文版

The amygdala is an important hub for regulating emotions and is involved in the pathophysiology of many mental diseases, such as depression and anxiety. Meanwhile, the endocannabinoid system plays a crucial role in regulating emotions and mainly functions through the cannabinoid type-1 receptor (CB1R), which is strongly expressed in the amygdala of non-human primates (NHPs). However, it remains largely unknown how the CB1Rs in the amygdala of NHPs regulate mental diseases. Here, we investigated the role of CB1R by knocking down the cannabinoid receptor 1 (CNR1) gene encoding CB1R in the amygdala of adult marmosets through regional delivery of AAV-SaCas9-gRNA. We found that CB1R knockdown in the amygdala induced anxiety-like behaviors, including disrupted night sleep, agitated psychomotor activity in new environments, and reduced social desire. Moreover, marmosets with CB1R-knockdown had up-regulated plasma cortisol levels. These results indicate that the knockdown of CB1Rs in the amygdala induces anxiety-like behaviors in marmosets, and this may be the mechanism underlying the regulation of anxiety by CB1Rs in the amygdala of NHPs.

Comparative study between S-(+)-ketamine­midazolam and fentanyl­droperidol in black-tufted marmosets (Callithrix penicillata) / Estudo comparativo entre S-(+)cetamina-midazolam e fentanil-droperidol em saguis-de-tufo-preto (Callithrix penicillata)

Sedative and antinociceptive effects of two anesthetic protocols in black-tufted marmosets were compared in this study. Twenty-six marmosets underwent chemical immobilization for physical examination, blood sampling, tattooing, and microchipping. Animals were randomly treated with S-(+)-ketamine (10 mg/kg) and midazolam (1 mg/kg) (KM) or fentanyl (12.5 µg/kg) and droperidol (625 µg/kg) (FD) given by intramuscular injection. Heart and respiratory rates were recorded. Sedation, antinociception, muscle relaxation, posture, auditory, and visual responses were evaluated using a scoring system. Sedation in KM was achieved faster (p < 0.001) and lasted for a shorter period of time (p = 0.0009). KM was similar to FD in its cardiorespiratory effects, auditory and visual responses. Both protocols promoted adequate sedation to allow manipulation. Animals in KM assumed lateral recumbency while animals in FD maintained a quadrupedal posture during evaluation. FD produced less intense sedation and muscle relaxation but a higher degree of antinociception compared to KM and is suitable for procedures that require analgesia in black-tufted marmosets.(AU)

O presente estudo comparou os efeitos cardiorrespiratórios, sedativos e antinociceptivos de dois protocolos anestésicos em saguis-de-tufo-preto (Callithrix penicillata). Vinte e seis saguis foram submetidos à contenção química para exame físico, coleta de sangue, tatuagem de identificação e microchip. Os animais foram tratados aleatoriamente com a associação de S-(+)-cetamina (10 mg/kg) e midazolam (1 mg/kg) (KM) ou fentanil (12,5 µg/kg) e droperidol (625 µg/kg) (FD), administrados por injeção intramuscular. Foram avaliadas frequência cardíaca, frequência respiratória, sedação, antinocicepção, relaxamento muscular, postura e resposta ao estímulo auditivo e visual. A sedação em KM foi alcançada mais rapidamente (p <0,001) e teve um tempo hábil mais curto (p = 0,0009). KM foi semelhante a FD nos efeitos cardiorrespiratórios, respostas auditivas e visuais. Os dois protocolos promoveram sedação adequada para manipulação. Os animais do grupo KM permaneceram em decúbito lateral durante a avaliação, enquanto os animais em FD mantiveram postura quadrupedal. FD resultou em sedação e relaxamento muscular de menor intensidade, porém com maior escore de antinocicepção em comparação com KM, sendo adequada para procedimentos que requerem analgesia em saguis-de-tufo-preto.(AU)

Platynosomum illiciens (Trematoda: Dicrocoeliidae) in a hybrid marmoset (Callithrix sp.) in the Municipality of Seropédica, RJ, Brazil - Case report / Platynosomum illiciens (Trematoda: Dicrocoeliidae) em um sagui híbrido (Callithrix sp.) no município de Seropédica, RJ, Brasil - Relato de caso

Abstract Platynosomum illiciens is a liver trematode encountered infecting mainly felids although it has also been reported in birds and in additional mammalian species, including non-human primates. The current study reports a natural P. illiciens infection primate of the genus Callithrix. The diagnosis was made using a combination of copro-parasitological techniques, morphological evaluation of adult specimens recovered from the liver during necropsy, and molecular analyses. Eggs were brown in color, oval, operculated, and contained a miracidium. Adult specimens recovered during necropsy were measured and showed dimensions compatible with P. illiciens. Molecular characterization of the trematode involved amplification by polymerase chain reaction (PCR), in combination with nucleotide sequencing, of an approximately 900 base pairs fragment corresponding to 18S-ITS1-5.8S ribosomal DNA. Sequenced amplicons showed 100% nucleotide identity with sequences deposited in the GenBank database as derived from specimens of P. illiciens recovered from cats in Malaysia and Brazil. It was concluded that the morphological and molecular analyses presented herein, confirmed the identification of the trematode recovered as P. illiciens.

Resumo Platynosomum illiciens é um trematódeo do fígado, encontrado principalmente em felinos, embora também tenha sido relatado em aves e outras espécies de mamíferos, incluindo primatas não humanos. Este estudo relata um caso de parasitismo natural por P. illiciens em um primata do gênero Callithrix. O diagnóstico foi feito com uma combinação de técnicas coproparasitológicas, avaliação morfológica de espécimes adultos recuperados do fígado durante a necropsia e análise molecular. Os ovos eram de cor marrom, ovais, operculados e continham um miracídio. Espécimes adultos recuperados durante a necropsia foram medidos e mostraram dimensões compatíveis com P. illiciens. A caracterização molecular do trematódeo envolveu a reação em cadeia da polimerase (PCR) e o sequenciamento de um fragmento de, aproximadamente, 900 pares de bases correspondentes ao DNA robosomal 18S-ITS1-5.8S, que apresentou 100% de identidade da sequência de nucleotídeos com as sequências do GenBank depositadas como derivadas de P. illiciens recuperados de gatos na Malásia e no Brasil, respectivamente. Concluiu-se pelas análises morfológicas e moleculares aqui apresentadas, que o trematódeo é da espécie P. illiciens.

Microbiota oral e retal de calitriquídeos (Callithrix sp) em área antropizada de Mata Atlântica, Rio de Janeiro, Brasil / [Oral and rectal microbiota of callitrichids (Callithrix sp) in an anthropized area of the Atlantic Forest in Rio de Janeiro, Brazil]

A proximidade dos primatas não humanos (PNH) com o ser humano pode ser considerada um fator de risco para transmissão de bactérias entre essas duas populações. Neste estudo, foi investigada a microbiota anfibiôntica aeróbica oral e retal de calitriquídeos em um fragmento de Mata Atlântica localizado no Rio de Janeiro, Brasil, e foram realizados testes fenotípicos para detecção de bactérias multirresistentes nos isolados encontrados. Foram capturados 14 calitriquídeos e coletadas 21 amostras (14 de cavidade oral e sete de cavidade retal) em dois pontos da mata próximos às habitações humanas. As espécies mais frequentes, na cavidade oral, foram Klebsiella oxytoca (50,0%), K. pneumoniae (28,6%), Kluyvera ascorbata (21,4%) e Stenotrophomonas maltophilia (21,4%) e, na cavidade retal, K. pneumoniae (85,7%), Escherichia coli (28,6%) e Enterobacter spp. (42,9%). Todos os 48 isolados da família Enterobacteriaceae foram negativos para ESBL (betalactamase de espectro ampliado), mostrando-se não produtores da enzima nos dois métodos utilizados: disco-aproximação e método de detecção automatizado. Na pesquisa de ERC (enterobactérias resistentes a carbapenêmicos), esses mesmos isolados não apresentaram resistência aos antibióticos imipenem, meropenem e ertapenem. Todas as bactérias isoladas apresentam um potencial zoonótico, o que representa um risco à saúde pública e à conservação das espécies.(AU)

Proximity of nonhuman primates (NHP) to humans can be considered a risk factor for transmission of pathogens between these two populations. This study investigated the oral and rectal aerobic bacterial microbiota of marmosets in an anthropized area of the Atlantic Forest located in Rio de Janeiro, Brazil, and performed phenotypic tests for detection of multidrug-resistant bacteria. Twenty-one samples (14 from the oral cavity and seven from the rectum) were collected from 14 Callithrix sp. captured in two sites of the forest near human dwellings. The most frequent species identified from the oral cavity swabs were Klebsiella oxytoca (50.0%), K. pneumoniae (28.6%), Kluyvera ascorbata (21.4%) and Stenotrophomonas maltophilia (21.4%), whereas the species most commonly identified from the rectum swabs were K. pneumoniae (85.7%), Enterobacter spp. (42.9%) and Escherichia coli (28.6%). All isolates of family Enterobacteriaceae showed no extended spectrum ß-lactamase production by disk-diffusion and automated detection tests. In the search for carbapenem-resistant enterobacteriaceae these isolates presented no resistance to the imipenem, meropenem and ertapenem antibiotics. The isolate of Staphylococcus aureus was susceptible to oxacillin and the isolate of Enterococcus was susceptible to vancomycin. All isolated bacteria showed zoonotic potential, thus posing a risk to species conservation and public health.(AU)

Rabies in Callithrix sp. in the urban area of Niterói, Rio de Janeiro, Brazil

Abstract In Brazil, rabies occurs mainly within an urban cycle, in which dogs and bats are reservoirs. This paper aims to report the occurrence of rabies in Callithrix sp. in Niterói, Rio de Janeiro, Brazil. In June 2019 a hybrid specimen was referred for diagnosis. The Direct Fluorescent Antibody, Mouse Inoculation, and Polymerase Chain Reaction tests were positive. A phylogenetic analysis was compatible with antigenic variant 3, characteristic of Desmodus rotundus. New studies should be undertaken to elucidate the real role of callitrichids in the urban rabies cycle.

First report of Gongylonema sp. in a free ranging callitrichid from the Brazilian Atlantic Forest: case report / Primeiro relato de Gongylonema sp. em um calitriquídeo de vida livre proveniente da Mata Atlântica brasileira: relato de caso

This is the first report of parasitism by Gongylonema sp. in a free-ranging callitrichid from the Atlantic forest of Espírito Santo, Brazil. A juvenile male of Geoffroy's marmoset (Callithrix geoffroyi) was euthanized due to poor prognosis, then necropsied. Samples of the tongue were collected for routine histological processing. Microscopically, there were transversal sections of adult nematodes within the epithelial layer of the mucosa of the tongue. Lingual scraping demonstrated a small number of oval embryonated eggs with a thick capsule. The morphology of the adult parasites and the eggs, associated with its location, were compatible with the Spirurida nematode of the genus Gongylonema. Further studies are needed to evaluate the impact of this parasite on free-ranging callitrichid populations.(AU)

Este é o primeiro relato de parasitismo por Gongylonema sp. em um calitriquídeo de vida livre proveniente da Mata Atlântica do Espírito Santo, Brasil. Um sagui-da-cara-branca (Callithrix geoffroyi), macho, jovem, foi eutanasiado, devido a prognóstico desfavorável, e necropsiado. Fragmentos de língua foram coletados para processamento histológico de rotina. Microscopicamente, havia cortes transversais de nematoides adultos na mucosa da língua. Na raspagem da mucosa da língua, foi observada pequena quantidade de ovos larvados ovais com cápsula espessa. A morfologia do parasita adulto e dos ovos, associada à localização do agente, é compatível com nematoide Spirurida do gênero Gongylonema. São necessários estudos adicionais para avaliar o impacto desse parasito nas populações de calitriquídeos de vida livre.(AU)

Nephroblastoma in a black-tufted marmoset (Callithrix penicillata) / Nefroblastoma em um sagui-de-tufos-pretos (Callithrix penicillata)

A renal nephroblastoma is described in a free-living black-tufted marmoset (Callithrix penicillata) in Central Brazil. The monkey was found dead and subjected to necropsy. Gross anatomic changes consisted of a ruptured left kidney, which was almost completely effaced by a white to yellow, partially encapsulated friable mass. The left ureter was distended due to obstruction by a red, spherical, 2mm in diameter friable mass. The urinary bladder was also distended. Histologically the renal and ureteral masses consisted of a triphasic embryonal neoplasm composed of embryonic epithelium forming glomeruli and tubules, polygonal blastemal cells, and a mesenchymal stroma. The embryonic epithelium exhibited rare nuclear immunoreactivity for WT-1, whereas blastemal cells exhibited robust cytoplasmic and rare nuclear immunoreactivity for WT-1; blastemal cells were also immunoreactive for vimentin. No immunoreactivity was detected for pan-cytokeratin (AE1/AE3), actin, and desmin. Morphological and immunohistochemical features of the present neoplasm are consistent with those described for renal nephroblastoma.(AU)

Descreve-se um caso de nefroblastoma maligno em um sagui de vida livre no Brasil Central. O macaco foi encontrado morto e encaminhado para necropsia. Na macroscopia, o rim esquerdo apresentava-se rompido e o parênquima estava substituído por um tecido neoplásico friável, parcialmente encapsulado e de superfície natural branca e de corte amarela. O ureter esquerdo apresentava-se distendido devido à obstrução por uma massa friável, vermelha, esférica, de 2mm de diâmetro. Histologicamente, as massas renal e ureteral consistiam de uma neoplasia embrionária composta por três populaçõies de células neoplásicas, composta por epitélio embrionário formando glomérulos e túbulos, células blastemais poligonais e um estroma mesenquimal. O epitélio embrionário exibiu imunorreactividade nuclear rara para WT-1, enquanto que as células blastemais exibiram imunorreactividade nuclear citoplasmática e rara para WT-1; As células blastemais também foram imunorreativas à vimentina. Nenhuma imunorreatividade foi detectada para pan-citoqueratina (AE1/AE3), actina e desmina. As características morfológicas e imuno-histoquímicas da presente neoplasia são consistentes com as descritas para o nefroblastoma renal.(AU)

Cytogenetics and Molecular Genetic Analysis of Chimerism in Marmosets (Callithrix: Primates)

ABSTRACT The birth of fraternal twins is a characteristic frequently observed in callitrichids. Cytogenetic studies have demonstrated hematopoietic chimerism in marmosets with the occurrence of two cell lines 2n=46,XX/46,XY in females and males co-twins, without phenotypic changes. Amplification by PCR have also been used to verify the presence of the SRY gene in female chimaeras. Our aim was to verify the occurrence of chimerism in Callithrix sp. individuals considered as hybrids according to their intermediate phenotypes between C. jacchus and C. penicillata. Blood samples from 37 Callithrix sp. individuals were collected. Hematopoietic chimerism 2n=46,XX/46,XY was detected by cytogenetic analysis in five individuals, three males and two females. A fragment of approximately 200bp of the SRY gene was amplified in seven females with normal external genitalia. The percentage of 32% of chimeric individuals detected in the present study is similar to that observed for pure specimens of Callithrix. These data suggests that hybridization probably does not interfere with the occurrence of twin gestation, nor of chimerism. Although cytogenetics is the main tool to identify the two cell lineages present in cases of chimerism, the amplification of the SRY gene by PCR has proved to be more efficient to identify the Y chromosome in cases of chimeric female marmoset.

Pathological and parasitological characterization of Prosthenorchis elegans in a free-ranging marmoset Callithrix geofroyi from the Brazilian Atlantic Forest / Caracterização patológica e parasitológica de Prosthenorchis elegans em Callithrix geoffroyi primate de vida livre da Mata Atlântica

Prosthenorchis elegans is an acanthocephalan intestinal parasite reported in neotropical primates. Despite parasitism by P. elegans having already been described in wild marmosets in the Brazilian Atlantic Forest, there are no reports of this infection in wild Geoffroy's marmoset (Callithrix geofroyi). The aim of this study is to report one case of P. elegans parasitism in a free-ranging C. geoffroyi from Brazilian Atlantic Forest in Espírito Santo state, and characterize the pathological and parasitological findings of this infection. One Geoffroy's marmoset necropsied at the Vila Velha University's Veterinary Pathology Laboratory presented intense chronic transmural ulcerative enteritis associated with twenty cylindrical helminths present in the jejunum and ileum. We can conclude that parasitism by P. elegans occurs in free-ranging groups of Geoffroy's marmosets. Its infection produced severe intestinal lesions even in free-ranging marmoset and therefore is a threat to this animal's survival in wildlife and can have some impact on primate conservation in the Brazilian Atlantic Forest.(AU)

Prosthenorchis elegans é um acantocéfalo intestinal descrito em primatas neotropicais. Apesar do parasitismo por P. elegans já ter sido descrito de saguis da Mata Atlântica brasileira, não há relatos da infecção em saguis-da-cara-branca (Callithrix geofroyi) de vida livre. O objetivo deste estudo é relatar um caso de parasitismo por P. elegans em um C. geoffroyi de vida livre proveniente da Mata Atlântica brasileira no Estado do Espírito Santo e caracterizar os achados patológicos e parasitológicos dessa infecção. Um sagui-da-cara-branca foi necropsiado no Laboratório de Patologia Veterinária da Universidade de Vila Velha, onde foi observada intensa enterite ulcerativa transmural crônica associada a vinte helmintos cilíndricos presentes no jejuno e íleo. Conclui-se que o parasitismo por P. elegans ocorre em grupos livres de saguis-da-cara-branca e a sua infecção leva a lesões intestinais graves; portanto, este parasita pode prejudicar a sobrevivência deste animal na vida selvagem e pode ter algum impacto na conservação de primatas na Mata Atlântica brasileira.(AU)

Origem dos nervos do plexo braquial de sagui-de-tufos-brancos (Callithrix jacchus Linnaeus, 1758) / Origin of brachial plexus nerves for common marmoset (Callithrix jacchus Linnaeus, 1758)

O sagui-de-tufos-brancos (Callithrix jacchus) é um pequeno primata que habita florestas arbustivas da Caatinga e a Mata Atlântica do Nordeste brasileiro. Essa espécie de sagui é muito comum e de fácil adaptação ao cativeiro, aspecto que estimula a captura clandestina desses animais, os quais são vítimas de maus tratos que causam lesões. A falta de conhecimento da anatomia de saguis e outros animais silvestres dificulta a aplicação segura de procedimentos clínicos, cirúrgicos e terapêuticos. O plexo braquial tem sido descrito em algumas espécies animais, porém até o momento não existia descrição em saguis-de-tufos-brancos (Callithrix jacchus). Para estudar o plexo branquial desses animais foram dissecados dez cadáveres de saguis-de-tufos-brancos (Callithrix jacchus) adultos sendo cinco machos e cinco fêmeas. Em todos os animais o plexo braquial originou- se dos nervos espinhais C5, C6, C7, C8 e T1, com poucas variações na origem e nas contribuições dos nervos, em ambos os antímeros, formando três troncos: cranial (C5 e C6), médio (C7) e caudal (C8 e T1).(AU)

The common marmoset (Callithrix jacchus) is a small primate that inhabits shrub forests of the Caatinga and the Atlantic Forest in Brazilian Northeast. This species of marmoset is very common and easy to adaptation in captivity, aspect that encourages illegal capture of these animals that are victims of mistreatments, causing injuries. The lack of knowledge in anatomy in marmosets and other wildlife hinders the safe application of clinical, surgical and therapeutic procedures. The brachial plexus has been described in some species of animals, but so far, there is no description for common marmoset (Callithrix jacchus). To study the brachial plexus of these animals were dissected ten corpses of common marmosets (Callithrix jacchus) adults, being five males and five females. In all animals, the brachial plexus was originated from spinal nerves C5, C6, C7, C8 and T1, with few variations in origin and the contributions of the nerves in both antimeres, forming three trunks: the cranial (C5 and C6), medium (C7) and caudal (C8 and T1).(AU)

Descrição anatômica do plexo braquial de Callithrix jacchus e Callithrix penicillata / Anatomic description of brachial plexus of Callithrix jacchus and Callithrix penicillata

Callithrix jacchus e Callithrix penicillata são primatas de pequeno porte cuja utilização como modelo anatômico tem se mostrado cada vez mais frequente, não somente pela praticidade no manuseio como facilidade no trato em criatório e sua taxa de reprodução. Este estudo teve como objetivo descrever os componentes dos plexos braquial em Callithrix jacchus e penicillata. Para tanto, três espécimes com aproximadamente 8 anos e 240 g foram fixados em solução de formaldeído a 10%, e posteriormente dissecados e fotodocumentados. O plexo braquial do Callithrix jacchus e penicillata originou-se dos nervos espinhais C5 a T1 constituindo os troncos cranial, médio e caudal. A composição do plexo braquial destes animais se assemelha ao de outros primatas, bem como a outros mamíferos.(AU)

Callithrix jacchus and Callithrix penicillata are small primates used as anatomic model, not only for convenience in handling as ease in regard to breeding and reproductive rate. The aim of this study was to describe the components of the brachial plexus in Callithrix jacchus and C. penicillata. Three specimens about 8 years old and weighing 240g were fixed in 10% formaldehyde and subsequently dissected and photodocumented. The brachial plexus of Callithrix jacchus and C. penicillata originates from the spinal nerves C5 to T1 in continuation of the cranial, medium and flow trunk. The composition of the brachial plexus of these animals is similar to the one of other primates ands other mammals.(AU)

Detection of neutralizing antibody to human adenovirus type 5 in marmosets / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct a recombinant human adenovirus type 5 (Ad5) expressing luciferase and GFP reporter gene and detect neutralizing antibodies against adenovirus type 5 in common marmosets (Callithrix jacchus) to provide basic laboratory data for evaluating adenovirus vaccines.</p><p><b>METHODS</b>Luciferase and GFP reporter genes from plasmid pHAGE-CMV-GFP were inserted into pDC315 to construct the recombinant adenovirus shutter plasmid pDC315-Luc-GFP. The shutter plasmid was co-transduced with pBHGlox(delta)E1,3Cre in 293A cell line to package the recombinant adenovirus rAd5/Luc/GFP. Three rounds of plaque formation experiment were performed to select the monoclonal adenovirus followed by purification with cesium chloride density gradient centrifugation and virus titration with TCID50 method. Chemiluminescence assay and flow cytometry were employed to detect the neutralizing antibody levels in 14 common marmosets.</p><p><b>RESULTS</b>The shuttle plasmid pDC315-Luc-GFP was successfully constructed and the recombinant adenovirus rAd5/Luc/GFP was packaged with a the titer reaching 6.9×10(11.5) PFU/mL. In the 14 marmosets, chemiluminescence assay identified 4 (28.6%) marmosets that were positive for Ad5-neutralizing antibodies, including 2 with a antibody titer of 1/16 and another 2 with a titer of 1/32; flow cytomery detected Ad5-neutralizing antibodies in 3 marmosets at the titer of 1/16.</p><p><b>CONCLUSION</b>Chemiluminescence assay is a simple, sensitive, and accurate modality for detecting Ad5-neutralizing antibodies. Common marmosets have a very low positivity rate for Ad5-neutralizing antibodies and are therefore promising models for studying adenovirus-based vaccines and therapies.</p>

Behavior, PET and Histology in Novel Regimen of MPTP Marmoset Model of Parkinson's Disease for Long-Term Stem Cell Therapy

Stem cell technologies are particularly attractive in Parkinson's disease (PD) research although they occasionally need long-term treatment for anti-parkinsonian activity. Unfortunately, 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) widely used as a model for PD has several limitations, including the risk of dose-dependent mortality and the difficulty of maintenance of PD symptoms during the whole experiment period. Therefore, we tested if our novel MPTP regimen protocol (2 mg/kg for 2 consecutive days and 1 mg/kg for next 3 consecutive days) can be maintained stable parkinsonism without mortality for long-term stem cell therapy. For this, we used small-bodied common marmoset monkeys (Callithrix jacchus) among several nonhuman primates showing high anatomical, functional, and behavioral similarities to humans. Along with no mortality, the behavioral changes involved in PD symptoms were maintained for 32 weeks. Also, the loss of jumping ability of the MPTP-treated marmosets in the Tower test was not recovered by 32 weeks. Positron emission tomography (PET) analysis revealed that remarkable decreases of bindings of ¹⁸F-FP-CIT were observed at the striatum of the brains of the marmosets received MPTP during the full period of the experiment for 32 weeks. In the substantia nigra of the marmosets, the loss of tyrosine hydroxylase (TH) immunoreactivity was also observed at 32 weeks following the MPTP treatment. In conclusion, our low-dose MPTP regimen protocol was found to be stable parkinsonism without mortality as evidenced by behavior, PET, and TH immunohistochemistry. This result will be useful for evaluation of possible long-term stem cell therapy for anti-parkinsonian activity.

Use of Nonhuman Primates for the Development of Bioengineered Female Reproductive Organs

Nonhuman primates (NHPs) have been widely used in reproductive biology, neuroscience, and drug development since a number of primate species are phylogenetically close to humans. In this review, we summarize the use of NHPs for nonclinical application in the reproductive system disorders including the loss or failure of an organ or tissue. Causes of infertility include congenital aplasia and acquired disorders of the reproductive organs. In addition, anti-cancer treatments can deplete ovarian follicles, leading to premature ovarian failure, infertility and long-term health risks. Along with a limited supply of human reproductive organs, anatomic/physiologic similarities to humans support the need for NHP models (New-World monkeys such as the common marmoset and Old-World monkeys such as cynomolgus and rhesus monkeys) to promote the advances in female infertility studies. For maintaining and executing animal studies using NHP, special protocols including animal care, anesthetic protocol, surgical technique, and immunosuppressive protocol are necessary. With a growing interest in the potential therapies such as endometrial tissue engineering, and ovary/follicle cryopreservation and grafting in Korea, this review can be useful in selecting appropriate animal models and can bridge between nonclinical studies and clinical applications by providing detailed information on the use of NHPs in the field of reproductive organ disorders.

Hepatic capillariasis in two captive Callithrix penicillata / Capilariose hepática em dois Callithrix penicillata de cativeiro

This study aimed to describe the occurrence of hepatic capillariasis in two non-human primates of the species Callithrix penicillata that were kept in captivity. Case 1 was in a young female, whose liver presented pale coloring, softening due to autolysis and several noticeably whitish areas. Case 2 was in a male of unknown age, whose liver was slightly pale and brownish. Histological analysis on the samples revealed extensive interstitial fibrosis with hemosiderotic areas. Adult nematodes of Capillaria hepatica and numerous eggs, morphologically characterized by their non-embryonic ellipsoid shape, with bipolar plugs that did not protrude from the shell, thick and birefringent, with a porous layer, thus giving a striated appearance to the eggs. Some cystic formations presented nematode eggs surrounded by numerous foreign-body giant cells. The literature consulted made little reference to reports of Capillaria hepatica in non-human primates. However, similar infections have been extensively studied in domestic rodents.(AU)

O presente trabalho relata a ocorrência de dois casos de capilariose hepática em primatas não-humanos da espécie Callithrix penicilatta mantidos em cativeiro. O caso 1 era uma fêmea jovem, que exibiu fígado de cor pálida, suavizada devido a autólise e algumas áreas múltiplas visivelmente esbranquiçadas. O caso 2 foi registrado em um macho adulto, de idade desconhecida, com fígado levemente pálido e acastanhado. A análise histológica das amostras revelou extensa fibrose intersticial com áreas de hemossiderose, exemplares de Capillaria hepatica adultos e numerosos ovos morfologicamente caracterizados pela sua forma elipsóide, não-embrionárias, com tampões bipolares que se sobressaem da casca, que é espessa, birrefringente e com uma camada porosa, dando uma aparência estriada ao ovo. Algumas formações císticas continham ovos do nematóide cercados por numerosas células gigantes do tipo corpo estranho. A literatura consultada possui poucas referências acerca da presença de Capillaria hepatica em primatas não-humanos. No entanto, infecções semelhantes são bem estudadas em roedores domésticos.(AU)

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

ABSTRACTRabies virus (RABV) isolated from different mammals seems to have unique characteristics that influence the outcome of infection. RABV circulates in nature and is maintained by reservoirs that are responsible for the persistence of the disease for almost 4000 years. Considering the different pattern of pathogenicity of RABV strains in naturally and experimentally infected animals, the aim of this study was to analyze the characteristics of RABV variants isolated from the main Brazilian reservoirs, being related to a dog (variant 2),Desmodus rotundus (variant 3), crab eating fox, marmoset, and Myotis spp. Viral replication in brain tissue of experimentally infected mouse was evaluated by two laboratory techniques and the results were compared to clinical evolution from five RABV variants. The presence of the RABV was investigated in brain samples by fluorescent antibody test (FAT) and real time polymerase chain reaction (qRT-PCR) for quantification of rabies virus nucleoprotein gene (N gene). Virus replication is not correlated with clinical signs and evolution. The pattern of FAT is associated with RABV replication levels. Virus isolates from crab eating fox and marmoset had a longer evolution period and higher survival rate suggesting that the evolution period may contribute to the outcome. RABV virus variants had independent characteristics that determine the clinical evolution and survival of the infected mice.

Evaluation of arboviruses of public health interest in free-living non-human primates (Alouatta spp., Callithrix spp., Sapajus spp.) in Brazil

INTRODUCTION: The aim of the present study was to evaluate the presence of arboviruses from the Flavivirus genus in asymptomatic free-living non-human primates (NHPs) living in close contact with humans and vectors in the States of Paraná and Mato Grosso do Sul, Brazil. METHODS: NHP sera samples (total n = 80, Alouatta spp. n = 07, Callithrix spp. n = 29 and Sapajus spp. n = 44) were screened for the presence of viral genomes using reverse transcription polymerase chain reaction and 10% polyacrylamide gel electrophoresis techniques. RESULTS: All of the samples were negative for the Flavivirus genome following the 10% polyacrylamide gel electrophoresis analysis. CONCLUSIONS: These negative results indicate that the analyzed animals were not infected with arboviruses from the Flavivirus genus and did not represent a risk for viral transmission through vectors during the period in which the samples were collected. .

Estudo endócrino reprodutivo e do comportamento sócio-sexual de sagui-de-tufo-preto (Callithrix penicillata) mantido em cativeiro / Reproductive endocrine and socio-sexual behavior study of black-tufted-marmoset (Callithrix penicillata) kept in captivity

A comunicação do estado reprodutivo nos primatas da família Callithrichidae, depende principalmente dos comportamentos sócio-sexuais como um sistema de sinalização primário, uma vez que nestas espécies a ovulação não é percebida pelos machos. Neste trabalho, os padrões de comportamentos sócio-sexuais foram analisados em conjunto com as concentrações de metabólitos fecais dos esteróides sexuais progesterona (MFP), estradiol (MFE) e testosterona (MFT) em casais cativos de Sagüi-de-tufos-pretos (Callithrix penicillata), nas diferentes fases do ciclo ovariano. O grupo estudado era composto por quarto casais adultos, mantidos no Centro de Reabilitação de Animais Selvagens da prefeitura de São Paulo. Os padrões comportamentais foram registrados pelo método de amostragem focal por intervalo de tempo a cada 30 segundos, cinco vezes por semana, totalizando 14.400 registros por animal. A mensuração das concentrações de metabólitos fecais dos esteroides sexuais foram realizados pelo método de enzima imunoensaio (EIE). Os resultados obtidos dessas concentrações possibilitaram a determinação endócrina das fases do ciclo ovariano (folicular e luteal) e de suas respectivas durações, assim como a determinação da fase periovulatória. Foram caracterizados 31 ciclos ovarianos completos, com duração de 24,3±4,1 dias (média ±DP), sendo que a fase folicular compreendeu 13,04±4,8dias e a fase lútea 11,2±4,2 dias. Os comportamentos sócio-sexuais (marcação por cheiro, cheirar genitália, catação e apresentação sexual) e a variável "proximidade" mostraram-se significativamente mais prevalentes na fase periovulatória do que nas demais fases do ciclo. Não houve alteração das concentrações de MFT dos machos ao longo de todo o período estudado. A análise conjunta das concentrações de metabólitos fecais de esteróides sexuais e dos comportamentos sócio-sexuais possibilitou um melhor entendimento das relações endócrino-comportamentais e reprodutivas de C. penicillata.

The communication of the female reproductive status in Callithrichidae relies mainly on the socio-sexual behavior, as generally the ovulation is concealed in this primate family by a primary signaling system. In this study the socio-sexual behavior patterns was analyzed in association with the concentration of fecal metabolites of sex steroid hormones progesterone (MFP), estradiol (MFE) and testosterone (MFT) in captive couples of Black-Tufted-Marmoset (Callithrix penicillata), during the different phases of the ovarian cycle. The studied group was composed of four adult couples kept in the São Paulo City Wild Animals Rehabilitation Center. The behavioral patterns were record by focal samplings, with 30 seconds intervals for each observation, five days a week, totalizing 14.400 registers per animal. The measurement of fecal metabolites of progesterone (MFP), estradiol (MFE) and testosterone (MFT) proceeded by enzyme immune assay (EIA). The results allowed to determine the duration of the ovarian cycle and to characterize three different phases (follicular, periovulatory and luteal). It was possible to determine 31 complete cycles that lasted 24.3±4.1 days (Mean ± SD). The follicular and luteal phases lasted 13.04±4.8 and the luteal phase 11.2±4.2 days. The behavioral patterns (scent marking, sniff genitals, grooming and sexual presentation) were more prevalent in the periovulatory phase as the behavioral variable "proximity" as well. There were no variations in the concentration of MFT in the males during the period studied. The associated analyses of the fecal metabolite of sex steroids and the socio-sexual behaviors led to a better understanding of the factors involved in the reproduction of C. penicillata.

Translocation and radio-telemetry monitoring of pygmy marmoset, Cebuella pygmaea (Spix, 1823), in the Brazilian Amazon / Translocação e monitoramento por radiotelemetria do mico-leãozinho, Cebuella pygmaea (Spix, 1823), na Amazônia brasileira

Two groups of pygmy marmoset (Cebuella pygmaea) were rescued along the left bank of the Madeira River during the formation of Santo Antônio Hydroelectric Dam reservoir in the state of Rondônia, Northern Brazil. Reintroduction of both groups occurred in areas of open Tropical rainforest located within the project´s Permanent Preservation Area. A post-release monitoring was conducted for three months using radio-telemetry. Individuals of each group remained together and settled in stable home ranges near their respective release sites. The mortality rate of translocated animals was about 7%. This seems to be the first report documenting the complete group translocation of C. pygmaea and the first to successfully employ radio-telemetry techniques in monitoring this species. This study demonstrated the feasibility of translocation and the use of radio-telemetry in monitoring C. pygmaea.

Dois grupos de mico leãozinho (Cebuella pygmaea) foram resgatados ao longo da margem esquerda do Rio Madeira durante o enchimento do reservatório da Usina Hidrelétrica Santo Antônio, em Rondônia, norte do Brasil. A reintrodução de ambos os grupos deu-se em áreas de floresta ombrófila densa aberta dentro da Área de Preservação Permanente do empreendimento. Após a reintrodução, empregou-se a técnica de radiotelemetria para o monitoramento dos animais por aproximadamente três meses. Cada um dos grupos manteve-se coeso e estabeleceu suas áreas de vida em regiões próximas àquelas onde foram soltos. A taxa de mortalidade foi de aproximadamente 7%. Este parece ser o primeiro trabalho a documentar a translocação de grupos completos de C. pygmaea e o primeiro a empregar com sucesso a radiotelemetria no monitoramento desta espécie. Este estudo, portanto, demonstra a factibilidade da translocação e do uso da radio-telemetria para monitorar C. pygmaea.

Modeling Parkinson's disease in the common marmoset (Callithrix jacchus): overview of models, methods, and animal care / 한국실험동물학회지

The common marmoset (Callithrix jacchus) is a small-bodied, popular New World monkey and is used widely in reproductive biology, neuroscience, and drug development, due to its comparative ease of handling, high reproductive efficiency, and its unique behavioral characters. In this review, we discuss the marmoset models in Parkinson's disease (PD), which is a neurological movement disorder primarily resulting from a degeneration of dopaminergic neurons with clinical features of tremor, rigidity, postural instability, and akinesia. The most common PD models involve the administration of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) or 6-hydroxydopamine to study the pathogenesis and to evaluate novel therapies. Following the systemic or local administration of these neurotoxins, the marmosets with very severe Parkinson's symptoms are recommended to be placed in an intensive care unit with artificial feeding to increase survival rate. All procedures with MPTP should be conducted in a special room with enclosed cages under negative-pressure by trained researchers with personal protection. Behavioral tests are conducted to provide an external measure of the brain pathology. Along with several biomarkers, including alpha-synuclein and DJ-1, non-invasive neuroimaging techniques such as positron emission tomography and magnetic resonance imaging are used to evaluate the functional changes associated with PD. With the recent growing interest in potential and novel therapies such as stem cell and gene therapy for PD in Korea, the marmoset can be considered as a suitable non-human primate model in PD research to bridge the gap between rodent studies and clinical applications.