PESQUISA VETERINÁRIA BRASILEIRA

Abstract in English:

ABSTRACT.- Sgai M.G.F.G., Pizzutto C.S., Viau P., Oliveira C.A. & Guimarães M.A.B.V. 2015. [Reproductive endocrine and socio-sexual behavior study of black-tufted-marmoset (Callithrix penicillata) kept in captivity.] Estudo endócrino reprodutivo e do comportamento sócio-sexual de sagui-de-tufo-preto (Callithrix penicillata) mantido em cativeiro. Pesquisa Veterinária Brasileira 35(3):304-310. Departamento de Reprodução Animal, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, Av. Prof. Dr. Orlando Marques de Paiva 87, Cidade Universitária, São Paulo, SP 055508-270, Brazil. E-mail: manuelafraga@hotmail.com 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.

• Black-tufted-marmoset Callithrix penicillata reproduction sagui-de-tufos-pretos reprodução

Abstract in Portuguese:

RESUMO.- Sgai M.G.F.G., Pizzutto C.S., Viau P., Oliveira C.A. & Guimarães M.A.B.V. 2015. [Reproductive endocrine and socio-sexual behavior study of black-tufted-marmoset (Callithrix penicillata) kept in captivity.] Estudo endócrino reprodutivo e do comportamento sócio-sexual de sagui-de-tufo-preto (Callithrix penicillata) mantido em cativeiro. Pesquisa Veterinária Brasileira 35(3):304-310. Departamento de Reprodução Animal, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, Av. Prof. Dr. Orlando Marques de Paiva 87, Cidade Universitária, São Paulo, SP 055508-270, Brazil. E-mail: manuelafraga@hotmail.com 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.

Abstract in English:

ABSTRACT.- Pizzutto C.S., Sgai M.G.F.G., Lopes D.A., Pessutti C., Nunes A., Furtado P.V., Oliveira C.A. & Guimarães M.A.B.V. 2015. Relation between the level of self-mutilation and the concentration of fecal metabolites of glucocorticoids in captive chimpanzees (Pan troglodytes). Pesquisa Veterinária Brasileira 35(1):62-66. Shape Brasil, Rua Dr. Miranda de Azevedo 779, Apartamento 104, Pompéia, São Paulo, SP 05027-000, Brazil. E-mail: cspizzutto@yahoo.com.br 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.

• Pan troglodytes self-mutilation glucocorticoid fecal metabolites automutilação glicocorticóides

Abstract in Portuguese:

RESUMO.- Pizzutto C.S., Sgai M.G.F.G., Lopes D.A., Pessutti C., Nunes A., Furtado P.V., Oliveira C.A. & Guimarães M.A.B.V. 2015. Relation between the level of self-mutilation and the concentration of fecal metabolites of glucocorticoids in captive chimpanzees (Pan troglodytes). [Relação do grau de automutilação com as concentrações de metabólitos fecais de glicocorticoides em chimpanzés (Pan troglodytes).] Pesquisa Veterinária Brasileira 35(1):62-66. Shape Brasil, Rua Dr. Miranda de Azevedo 779, Apartamento 104, Pompéia, São Paulo, SP 05027-000, Brazil. E-mail: cspizzutto@yahoo.com.br A influência do estresse de um ambiente nas variáveis endócrino-comportamentais de primatas vem sendo cada vez mais estudada por diversos autores, e mostram que comportamentos anormais associados a aumentos de glicocorticóides podem estar diretamente relacionados ao comprometimento do bem-estar. Neste trabalho foram utilizados 22 chimpanzés (Pan troglodytes) adultos, sendo 11 machos e 11 fêmeas mantidos em cativeiro de três instituições diferentes. Todos os animais tiveram seus comportamentos registrados pelo método de amostragem focal por intervalo de tempo, durante seis meses, totalizando 4800 registros para cada animal. Amostras fecais foram coletadas três vezes por semana, durante este período, para extração e dosagem de metabólitos fecais de glicocorticoides por radioimunoensaio. Os comportamentos estereotipados representaram 13,45+2,76% do total observado, sendo que dentre estes comportamentos a automutilação representou 38,28+3,98%. Os animais foram classificados em três graus diferentes, de acordo com o percentual da superfície corpórea com alopecia decorrente da automutilação. Foi encontrada uma correlação positiva de intensidade forte entre os graus de alopecia decorrentes de mutilação observados e as médias de concentrações de metabólitos fecais de glicocorticoides. Este resultado sugere fortemente que esta graduação de automutilação de um chimpanzé possa ser utilizada como uma ferramenta auxiliar importante nas avaliações das condições de adaptação do animal ao cativeiro, atuando como um indicador indireto da presença de estresse crônico.

Abstract in English:

ABSTRACT.- Vasconcellos A.S., Chelini M.O.M., Palme R., Guimarães M.A.B.V., Oliveira C.A. & Ades C. 2011. Comparison of two methods for glucocorticoid evaluation in maned wolves. Pesquisa Veterinária Brasileira 31(Supl.1):79-83. Department of Experimental Psychology, Institute of Psychology, University of São Paulo, Av. Professor Mello de Moraes 1721, São Paulo, SP 05508-030, Brazil. E-mail: angelv@usp.br Analysis of faecal glucocorticoid metabolites provides a powerful noninvasive tool for monitoring adrenocortical activity in wild animals. However, differences regarding the metabolism and excretion of these substances make a validation for each species and sex investigated obligatory. Although maned wolves (Chrysocyon brachyurus) are the biggest canids in South America, their behaviour and physiology are poorly known and they are at risk in the wild. Two methods for measuring glucocorticoid metabolites in maned wolves were validated: a radio- and an enzyme immunoassay. An ACTH challenge was used to demonstrate that changes in adrenal function are reflected in faecal glucocorticoid metabolites. Our results suggest that both methods enable a reliable assessment of stress hormones in maned wolves avoiding short-term rises in glucocorticoid concentrations due to handling and restraint. These methods can be used as a valuable tool in studies of stress and conservation in this wild species.

• ACTH corticosterone cortisol corticosterona

Abstract in Portuguese:

RESUMO.- Vasconcellos A.S., Chelini M.O.M., Palme R., Guimarães M.A.B.V., Oliveira C.A. & Ades C. 2011. Comparison of two methods for glucocorticoid evaluation in maned wolves. [Comparação de dois métodos para avaliação de glicocorticóides em lobos guarás.] Pesquisa Veterinária Brasileira 31(Supl.1):79-83. Department of Experimental Psychology, Institute of Psychology, University of São Paulo, Av. Professor Mello de Moraes 1721, São Paulo, SP 05508-030, Brazil. E-mail: angelv@usp.br A análise de metabólitos de glicocorticóides fecais é uma ferramenta não-invasiva poderosa para o monitoramento da atividade adrenal em animais silvestres. Entretanto, diferenças no metabolismo e na excreção dessas substâncias tornam obrigatória a validação desse método para cada espécie e sexo investigado. Embora os lobos guarás (Chrysocyon brachyurus) sejam os maiores canídeos da América do Sul, seu comportamento e fisiologia são ainda pouco conhecidos e estes animais são ameaçados. Dois métodos para mensuração de metabólitos de glicocorticóides em lobos guarás foram validados: um radio- e um enzima- imuno-ensaio. Um desafio de ACTH foi usado para demonstrar que alterações no funcionamento adrenal podem ser detectadas nas concentrações metabólitos de glicocorticóides fecais. Nossos resultados sugerem que ambos os métodos possibilitam uma avaliação confiável dos hormônios relacionados ao estresse em lobos guarás, evitando assim aumentos de curta duração nas concentrações glicocorticóides, devidos ao manejo e contenção. Esses métodos podem ser usados como uma ferramenta valiosa em estudos de estresse e conservação com essa espécie silvestre.

Abstract in English:

ABSTRACT.- Dias E.A., Nichi M. & Guimarães M.A.B.V. 2008. Comparison of two commercial kits and two extraction methods for fecal glucocorticoid analysis in ocelots (Leopardus pardalis) submitted to ACTH challenge. Pesquisa Veterinária Brasileira 28(7):329-334. Departamento de Reprodução Animal, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, Av. Prof. Dr. Orlando Marques de Paiva 87, Cidade Universitária, São Paulo, SP 05508-270, Brazil. E-mail: eantdias@usp.br The ocelot (Leopardus pardalis) is included in list of wild felid species protected by CITES and is part of conservation strategies that necessarily involve the use of assisted reproduction techniques, which requires practical and minimally invasive techniques of high reproducibility that permit the study of animal reproductive physiology. The objective of this study was to compare and validate two commercial assays: ImmuChem Double Antibody Corticosterone 125I RIA from ICN Biomedicals, Costa Mesa, CA, USA; and Coat-a-Count Cortisol 125I RIA from DPC, Los Angeles, CA, USA, for assessment of fecal glucocorticoid metabolites in ocelots submitted to ACTH (adrenocorticotropic hormone) challenge. Fecal samples were collected from five ocelots kept at the Brazilian Center of Neotropical Felines, Associação Mata Ciliar, São Paulo, Brazil, and one of the animals was chosen as a negative control. The experiment was conducted over a period of 9 days. On day 0, a total dose of 100 IU ACTH was administered intramuscularly. Immediately after collection the samples were stored at -20C in labeled plastic bags. The hormone metabolites were subsequently extracted and assayed using the two commercial kits. Previously it was performed a trial with the DPC kit to check the best extraction method for hormones metabolites. Data were analyzed with the SAS program for Windows V8 and reported as means ± SEM. The Schwarzenberger extraction method was slightly better when compared with the Wasser extraction method (103,334.56 ± 19,010.37ng/g of wet feces and 59,223.61 ± 12,725.36ng/g of wet feces respectively; P=0,0657). The ICN kit detected an increase in glucocorticoid metabolite concentrations in a more reliable manner. Metabolite concentrations (ng/g wet feces) on day 0 and day 1 were 66,956.28 ± 36,786.93 and 92,991.19 ± 28,555.63 for the DPC kit, and 205,483.32 ± 83,811.32 and 814,578.75 ± 292,150.47 for the ICN kit, respectively. The limit of detection for the ICN kit was 7.7 ng/mL for 100% B/Bo (25ng/mL for 88%B/Bo) and for the DPC kit it was 0.2ug/dL for 90.95% B/Bo (1ug/dL for 81.27% B/Bo). In conclusion it was confirmed that the Schwarzenberger extraction method and the ICN kit are superior for extracting and measuring fecal glucocorticoid metabolites in ocelot fecal samples.

• Stress Leopardus pardalis ocelot ACTH challenge fecal glucocorticoid jaguatirica ACTH glicocorticóides fecais

Abstract in Portuguese:

ABSTRACT.- Dias E.A., Nichi M. & Guimarães M.A.B.V. 2008. Comparison of two commercial kits and two extraction methods for fecal glucocorticoid analysis in ocelots (Leopardus pardalis) submitted to ACTH challenge. Pesquisa Veterinária Brasileira 28(7):329-334. Departamento de Reprodução Animal, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, Av. Prof. Dr. Orlando Marques de Paiva 87, Cidade Universitária, São Paulo, SP 05508-270, Brazil. E-mail: eantdias@usp.br The ocelot (Leopardus pardalis) is included in list of wild felid species protected by CITES and is part of conservation strategies that necessarily involve the use of assisted reproduction techniques, which requires practical and minimally invasive techniques of high reproducibility that permit the study of animal reproductive physiology. The objective of this study was to compare and validate two commercial assays: ImmuChem Double Antibody Corticosterone 125I RIA from ICN Biomedicals, Costa Mesa, CA, USA; and Coat-a-Count Cortisol 125I RIA from DPC, Los Angeles, CA, USA, for assessment of fecal glucocorticoid metabolites in ocelots submitted to ACTH (adrenocorticotropic hormone) challenge. Fecal samples were collected from five ocelots kept at the Brazilian Center of Neotropical Felines, Associação Mata Ciliar, São Paulo, Brazil, and one of the animals was chosen as a negative control. The experiment was conducted over a period of 9 days. On day 0, a total dose of 100 IU ACTH was administered intramuscularly. Immediately after collection the samples were stored at -20C in labeled plastic bags. The hormone metabolites were subsequently extracted and assayed using the two commercial kits. Previously it was performed a trial with the DPC kit to check the best extraction method for hormones metabolites. Data were analyzed with the SAS program for Windows V8 and reported as means ± SEM. The Schwarzenberger extraction method was slightly better when compared with the Wasser extraction method (103,334.56 ± 19,010.37ng/g of wet feces and 59,223.61 ± 12,725.36ng/g of wet feces respectively; P=0,0657). The ICN kit detected an increase in glucocorticoid metabolite concentrations in a more reliable manner. Metabolite concentrations (ng/g wet feces) on day 0 and day 1 were 66,956.28 ± 36,786.93 and 92,991.19 ± 28,555.63 for the DPC kit, and 205,483.32 ± 83,811.32 and 814,578.75 ± 292,150.47 for the ICN kit, respectively. The limit of detection for the ICN kit was 7.7 ng/mL for 100% B/Bo (25ng/mL for 88%B/Bo) and for the DPC kit it was 0.2ug/dL for 90.95% B/Bo (1ug/dL for 81.27% B/Bo). In conclusion it was confirmed that the Schwarzenberger extraction method and the ICN kit are superior for extracting and measuring fecal glucocorticoid metabolites in ocelot fecal samples.