Comparison of two commercial kits and two extraction methods for fecal glucocorticoid analysis in ocelots (Leopardus pardalis) submitted to ACTH challenge

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.

Comparison of methods for determination of nutrient digestibility of a dry kibble diet for ocelots

In this experiment, methods of total fecal collection (TFC) and internal markers (acid-insoluble ash - AIA, crude fiber - CF, and acid-detergent fiber - ADF) were compared for determination of the coefficients of apparent digestibility (CAD) for dry matter (DM), crude protein (CP), ether extract (EE), nitrogen-free extracts (NFE), and gross energy (GE) of commercial feline dry kibble for ocelots (Leopardus pardalis). Six adult animals, weighing 12.45±1.37 kg, gradually received experimental kibble in their usual diet until the beginning of the experiment and were submitted to an adaptation period ten days prior to the collection period. CAD obtained by TFC, AIA, CF, and ADF were, respectively, 73.7, 76.83, 62.01, and 46.03% for dry matter; 81.9, 84.8, 75.8, and 63.8% for crude protein; 85, 86.7, 78.5, and 69.1% for ether extract; 78.52, 79.55, 69.11, and 53.04% for nitrogen-free extracts; and 80.5, 82.2, 71.4, and 58.4% for gross energy. The AIA method showed to be efficient in determining coefficients of apparent digestibility and may contribute to investigations on the digestibility of diets for wild felines. In comparison to the items of ocelot's usual diet, the kibble used in this paper provided an adequate nutritional supply with reduced daily costs per animal.

Diets of ocelots (Leopardus pardalis), margays (L-wiedii), and oncillas (L-tigrinus) in the Atlantic rainforest in southeast Brazil

Felids play an important role in structuring the prey community, and thus in the dynamics of the plant community in tropical forests. The diets of three species of small cats, Leopardus pardalis (ocelot), L. wiedii (margay), and L. tigrinus (oncilla) were investigated by analysis of scats and regurgitations during a one-year study at Parque Estadual Serra do Mar, Nucleo Santa Virginia, southeastern Brazil. Small non-volant mammals were the most frequent prey in all the diets. The diets of the three cats were similar but although they consumed the same prey items, they showed differential use of them. Ocelots consumed more reptiles than the other species, oncillas preyed on birds and arboreal marsupials, whereas margays were generalists.

Effects of tiletamine/zolazepam-romifidine-atropine in ocelots (Leopardus pardalis)

Objectives: To evaluate the effects of a combination of tiletamine-zolazepam-romifidine-atropine in ocelots. Design: Prospective experimental trial. Animals: Eight captive adult ocelots (three females and five males). Methods: Calculated doses of tiletamine-zolazepam (3.75 mg kg -1), romifidine (50 μg kg-1) and atropine (0.04 mg kg-1) were administered intramuscularly. After immobilization, animals were weighed and the real doses determined. Heart rate, respiratory frequency, noninvasive systolic, diastolic, and mean arterial pressure, arterial oxygen hemoglobin saturation, and rectal temperature were measured. Data were analyzed by means of ANOVA for repeated measures, followed by the Tukey test to compare values over time. Results: Doses administered were 3.4 ± 0.6 mg kg-1 of tiletamine-zolazepam, 0.04 ± 7.0 mg kg-1 of romifidine, and 0.03 ± 0.007 mg kg-1 of atropine. The mean time to recumbency and duration of immobilization were 7.0 ± 4.5 and 109.2 ± 27.9 minutes, respectively. The median times to standing and walking were 52.3 [0-90] and 2.3 [0-69.3] minutes, respectively. A decrease in heart rate was observed 45 minutes following drug administration. Arterial blood pressure was maintained during the study. Conclusions and clinical relevance: This protocol produced good immobilization in ocelots with minimal changes over time in cardiovascular parameters.

HEMOPLASMAS IN WILD CANIDS AND FEUDS IN BRAZIL

Hemotropic mycoplasmas, epicellular erythrocytic bacterial parasites lacking a cell wall, are the causative agents of infectious anemia in numerous mammalian species. The presence of hemotropic mycoplasmas in blood samples of neotropical and exotic wild canids and felids from Brazilian zoos were recorded using molecular techniques. Blood samples were collected from 146 Brazilian wild felids, 19 exotic felids, 3 European wolves (Canis lupus), and from 97 Brazilian wild canids from zoos in the Brazilian states of Sao Paulo and Mato Grosso and the Federal District. Using conventional polymerase chain reaction (PCR), this work found 22 (13%) wild felids positive to Candidatus Mycoplasma haemominutum [4 jaguars (Panthera onca); 3 pumas (Puma concolor); 10 ocelots (Leopardus pardalis); 2 jaguarondis (Puma yagouaroundi); and 3 little spotted cats (Leopardus tigrinus)]. Only one little spotted cat (Leopardus tigrinus) was positive to Mycoplasma haemofelis, and none was positive to Candidatus Mycoplasma turicensis. Two bush dogs (Speothos venaticus) were positive for a Mycoplasma sp. closely related to Candidatus Mycoplasma haematoparvum, and two European wolves were positive for a Mycoplasma sp. closely related to candidatus Mycoplasma haemominutum. This is the first study regarding the molecular detection of hemotropic mycoplasmas in wild canids.

Serological survey of Toxoplasma gondu in captive Neotropical felids from Southern Brazil

Toxoplasma gondu is the causative intracellular protozoan of toxoplasmosis inhuman being and animals Members of the Felidae family are considered the single definitive host for the infection both wild and domestic cats are able to excrete oocysts in the environment Wild cats maintained in captivity may serve as source of infection for other clinically susceptible animals in the same environment The aim of this study was to determine the frequency of T gondu IgG antibodies in 57 neotropical felids (1 Leopardus geoffroyi 3 Puma yagouaroundi 17 Leopard us wiedu 22 Leopardus tigrinus and 14 Leopard us pardalis) kept at the Bela Vista Biological Sanctuary Itaipu Binacional Southern Brazil by the modified agglutination test (MAT) using titer 16 as cut-off point Seropositivity was observed in 38/57 (66 67% 95% CI 53 66-77 51%) samples with higher frequency in ocelots (71 43%) Wild-caught felids were three times more likely to be infected when compared to zoo-born animals (P <= 0 05) and age of wild-caught animals (P= 0 6892 95% CI = 0 7528-166) was not significant as a risk factor for the infection the same occurring with zoo-born animals (P=0 05 95% CI = 06267-24052) These results suggest that despite efforts to control T gondu infection in zoo facilities such as individual pens hygiene monitoring veterinary care and pre-frozen meat offered as food non-domestic feuds kept in captivity particularly the wild-caught specimens may be invariably exposed to infection due to other environmental sources (C) 2010 Elsevier B V All rights reserved

Density estimates and conservation of Leopardus pardalis southernmost population of the Atlantic Forest

ABSTRACT Using camera traps and capture/recapture analyses we recorded the presence and abundance of cat species at Turvo State Park, in southern Brazil. Ocelot [Leopardus pardalis (Linnaeus, 1758)] population density was estimated for two areas of the park, with differing management profiles. Density estimates varied from 0.14 to 0.26 indiv. km2. Another five cat species were recorded at very low frequencies, precluding more accurate analyses. We estimate 24 to 45 ocelots occur in the reserve, which is probably too small for long-term maintenance of the population, if isolated. However, if habitat integrity and connectivity between the Park and the Green Corridor of Misiones is maintained, an estimated ocelot population of 1,680 individuals should have long-term viability.

Germ Cell Transplantation in Felids: A Potential Approach to Preserving Endangered Species

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

MOLECULAR and SEROLOGICAL DETECTION of BABESIA SPP. IN NEOTROPICAL and EXOTIC CARNIVORES IN BRAZILIAN ZOOS

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

Molecular detection of tick-borne bacterial agents in Brazilian and exotic captive carnivores

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

SEROLOGIC SURVEY FOR LEPTOSPIRA SPP. IN CAPTIVE NEOTROPICAL FELIDS IN FOZ do IGUACU, PARANA, BRAZIL

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

Molecular detection of Cytauxzoon spp. in asymptomatic Brazilian wild captive felids

Cytauxzoon. spp. DNA was detected for the first time in blood samples from asymptomatic Brazilian wild captive felids. In 2606, 72 EDTA blood samples from seven wild felids species: Puma concolor (pinna), Leopardus pardalis (ocelot), Puma yagouaroundi (jaguarundi), Leopardus wiedii (margay), Leopardus tigrinus (little spotted cat), Oncifelis colocolo (pampas cat) and Panthera. onca. (jaguar) were analyzed using polymerase chain reaction to amplify the 18S rRNA gene segment in order to verify the presence of Cytauxzoon spp. DNA. Nine samples were positive: six ocelots, two pumas, and one jaguar. In Brazil, wild felids may be natural reservoirs for Cytauxzoon spp.

Serological survey of Toxoplasma gondii in captive Neotropical felids from Southern Brazil

Toxoplasma gondii is the causative intracellular protozoan of toxoplasmosis in human being and animals. Members of the Felidae family are considered the single definitive host for the infection; both wild and domestic cats are able to excrete oocysts in the environment. Wild cats maintained in captivity may serve as source of infection for other clinically susceptible animals in the same environment. The aim of this study was to determine the frequency of T. gondii IgG antibodies in 57 neotropical felids (1 Leopardus geoffroyi; 3 Puma yagouaroundi; 17 Leopardus wiedii; 22 Leopardus tigrinus; and 14 Leopardus pardalis) kept at the Bela Vista Biological Sanctuary, Itaipu Binacional, Southern Brazil, by the modified agglutination test (MAT) using titer 16 as cut-off point. Seropositivity was observed in 38/57 (66.67%; 95% CI 53.66-77.51%) samples, with higher frequency in ocelots (71.43%). Wild-caught felids were three times more likely to be infected when compared to zoo-born animals (P≤ 0.05) and age of wild-caught animals (P= 0.6892; 95% CI. = 0.7528-1.66) was not significant as a risk factor for the infection, the same occurring with zoo-born animals (P= 0.05; 95% CI. = 0.6267-24.052). These results suggest that, despite efforts to control T. gondii infection in zoo facilities, such as individual pens, hygiene monitoring, veterinary care and pre-frozen meat offered as food, non-domestic felids kept in captivity, particularly the wild-caught specimens, may be invariably exposed to infection due to other environmental sources. © 2010 Elsevier B.V.

Parâmetros eletrocardiográficos de felideos selvagens da Amazônia (Panthera onca, Leopardus pardalis, Leopardus wiedii, Herpailurus yaguarondi) criados em cativeiro no estado do Pará, tranquilizados com ketamina e cloridrato de xilazina

Pretendeu-se estabelecer os parâmetros eletrocardiográficos (ECG) para felinos selvagens da Amazônia - Panthera onca (onça pintada), Leopardus pardalis (jaguatirica), Leopardus wiedii (gato-maracajá) e Herpailurus yaguarondi (gato-mourisco). Dentre estes felinos, 17 animais pertenciam ao Parque Zoobotânico Dr. Adhemar Monteiro (Capitão Poço/PA) e 2 animais ao 2° Batalhão de Infantaria de Selva (Belém/PA). Embora, o exame eletrocardiográfico não cause dor aos animas, é necessária a realização de contenção química, visto que os animais são perigosos e estressados. Dessa forma, instaurou-se um protocolo de contenção química, visando evitar fugas, estresse excessivo e proteger a integridade física dos animais e do examinador/auxiliares. Antes de serem sedados os animais passaram por jejum alimentar de 24 horas e hídrico de 12 horas. Posteriormente, foram contidos quimicamente, através do uso associativo entre os anestésicos cloridrato de ketamina (10mg/Kg) e cloridrato de xilazina (1mg/Kg), utilizando dardos com auxílio de uma zarabatana ou aplicação intra-muscular após contenção física com puçá. Na realização do eletrocardiograma utilizou-se o mesmo protocolo usado para os animais domésticos. A média de batimentos cardíacos foram os seguintes: 90 bpm para jaguatiricas, 106 bpm gato-maracajá, 166 bpm gato-mourisco e 91 bpm onças pintadas. A maioria das espécies apresentaram ritmo sinusal normal. O eixo elétrico médio variou de 90° a 120°. Os complexos QRS foram predominantemente positivos em DI, DII, DIII e AVF, e negativos em AVR e AVL. As derivações pré-cordiais apresentaram resultados semelhantes aos esperados para o gato doméstico. Os parâmetros observados durantes este estudo, correlacionados com os animais domésticos, apresentaram algumas diferenças, que se devem em função das diferenças fisiológicas associadas ao maior tamanho corporal dos felinos selvagens. Este é um estudo pioneiro visando solucionar problemas com relação ao ECG de felinos selvagens da Amazônia. Portanto, investigações mais amplas sobre o mesmo tema são necessárias para estabelecer critérios a respeito de anormalidades nessas espécies, devendo incluir outras drogas anestésicas e relatórios para combinações de características eletrocardiográficas de animais com doença cardíaca e com distúrbios eletrolíticos.

Intraspecific, interspecific, and seasonal differences in the diet of three mid-sized carnivores in a large neotropical wetland

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)