Guard-hair microstructure of Brazilian felids: considerations for species identification

O exame da microestrutura da cutícula e medula dos pêlos é uma técnica simples e de baixo custo para identificar espécies de mamíferos para uma variedade de aplicações. Foram estudados pêlos-guarda de 66 indivíduos de oito espécies de felídeos brasileiros (Leopardus colocolo, L.geoffroyi, L.pardalis, L.tigrinus, L.wiedii, Panthera onca, Puma concolor, Puma yagouaroundi), através de amostras colhidas de animais anestesiados em zoológicos. A microestrutura dos pêlos-guarda foi analisada e descrita através de impressões cuticulares e preparações medulares, e posteriormente foi feito um teste cego para avaliar a acurácia da identificação específica. Embora tenham sido encontrados caracteres morfológicos distintos para cada espécie, a sutileza destes caracteres e sua sobreposição entre as diferentes espécies resultaram em uma acurácia relativamente baixa (75%). A identificação de pares ou trios de espécies cujos pêlos têm morfologia mais semelhante (Grupo A: L. pardalis, L. tigrinus, L. wiedii; Grupo B: L. colocolo, L. geoffroyi, P. yagouaroundi; Grupo C: P. concolor, P. onca) elevou a acurácia significativamente (91%). A identificação dos felídeos brasileiros através da microestrutura de seus pêlos é desafiadora e requer um exame cuidadoso de caracteres sutis, devendo ser apoiada por outras técnicas mais acuradas e/ou ser limitada principalmente às aplicações que não requerem identificação específica ou que trabalhem em escalas taxonômicas mais amplas.The analysis of cuticle and medulla hair microstructure is a simple and inexpensive technique to identify mammal species for a variety of applications. We studied the guard-hairs of 66 individuals of eight felid species occurring in Brazil (Leopardus colocolo, L.geoffroyi, L.pardalis, L.tigrinus, L.wiedii, Panthera onca, Puma concolor, Puma yagouaroundi), through hair samples collected from anesthetized zoo animals. The microstructure of the guard-hairs was analyzed and described through cuticle impressions and medulla preparations; a blind test was conducted to evaluate the accuracy of species identification. Although distinctive morphological characters could be identified for each species, the subtlety of these characters and the overlap of features among species resulted in a relatively poor accuracy (75%). The identification of pairs or trios of species whose hair has similar morphologies (Group A: L. pardalis, L. tigrinus, L. wiedii; Group B: L. colocolo, L. geoffroyi, P. yagouaroundi; Group C: P. concolor, P. onca) significantly improved accuracy (91%). The identification of Brazilian felids through the microstructure of their hair is challenging and requires careful examination of subtle features, and should be complemented by more accurate techniques and/or be limited mostly to applications where high accuracy is not essential or where a broader taxonomic scale is being evaluated

Contrafreeloading in maned wolves: Implications for their management and welfare

Contrafreeloading occurs when animals spend time and effort to obtain food in the presence of freely available food. There are several interpretations for such an apparent contradiction to optimal foraging models, with an emphasis either on the need to gather and update information about the environment or on the value of performing species-typical responses. Evidence suggests that both gathering information about the environment and the expression of species-typical behaviour are important for the welfare of captive animals. The aim of the present study was to assess the existence of contrafreeloading in maned wolves (Chrysocyon brachyurus), in a situation where animals could get food directly from a "free" source and/or search and handle hidden food items, an alternative that requires more effort and is probably more similar to natural foraging conditions. Eight captive, pair-housed maned wolves were given weekly choice tests in which they could obtain food either by approaching the usual food tray in one section of the enclosure (Tray), and/or by searching for food at variable sites amongst the vegetation in the other section of the enclosure (Scattered). Results indicate that maned wolves spent more time in the Scattered than in the Tray section of the enclosure (P = 0.02) and that they obtained about half of the food from that section (48.54% +/- SE 0.69). Our results, the first to demonstrate contrafreeloading in maned wolves, have implications for the husbandry and welfare of this endangered species. (C) 2012 Elsevier B.V. All rights reserved.FAPESP - Sao Paulo Research Foundation [2009/14258-4

Vaginal cytology for estrus detection in ocelot (Leopardus pardalis)

Utilizou-se a citologia vaginal por meio de diferentes métodos de coloração para detecção de cio em jaguatirica, Leopardus pardalis, pela estimulação hormonal exógena e pela avaliação das estruturas ovarianas por videolaparoscopia. Cinco fêmeas foram tratadas com eCG/hCG e FSH/LH a cada quatro meses pelo período de dois anos. Videolaparoscopia foi realizada após cada tratamento utilizando-se cetamina-xilazina e isoflurano. Esfregaços vaginais foram obtidos 15 dias antes e após a videolaparoscopia. As lâminas foram analisadas ao microscópio de luz quanto aos tipos celulares predominantes. Todos os animais apresentaram folículos maduros (>2mm) e corpos lúteos recentes em todas as intervenções. Não houve diferença significativa entre os resultados obtidos na mesma coloração de acordo com os tratamentos utilizados. Todas as técnicas mostraram-se eficientes na detecção de células superficiais queratinizadas anucleadas e nucleadas, intermediárias, parabasais e basais. Foi possível determinar a fase de estro em Leopardus pardalis por meio da citologia vaginalVaginal cytology was evaluated for estrus detection using different stains after hormonal stimulation with exogenous gonadotrophin (eCG/hCG, FSH/LH) and videolaparoscopy for ovarian structure evaluation. Five L. pardalis were treated four times during two years. After each treatment, videolaparoscopy was performed using Ketamine-Xylazine and Isoflurane. Vaginal cytology was made 15 days before and after videolaparoscopy. Three stains were used: Diff Quick, Papanicolaou, and Shorr. The slides were analyzed for the typical cell predominance. All the animals showed mature follicles (>2mm) and recent corpus luteum in all procedures. No significative difference was observed between the results in the same stain according to the treatment eCG/hCG and FSH/LH. All stains were efficient in detection of nucleated and anuclear superficial keratinized cells; intermediated, parabasal, and basal cells. Vaginal cytology can be used for estrus detection in Leopardus pardalisFAPES

Análise citogenética de oócitos de jaguatirica (Leopardus pardalis) e gato-do-mato-pequeno (Leopardus tigrinus) coletados após estimulação ovariana

Este estudo representa a primeira avaliação da maturação nuclear de oócitos por análise citogenética realizada em duas espécies de felídeos brasileiros ameaçados de extinção: L. pardalis (n=5) e L. tigrinus (n=4). Os animais foram submetidos à estimulação ovariana alternada com eCG-hCG e pFSH-pLH a cada quatro meses pelo período de dois anos, perfazendo um total de 6 intervenções. Os oócitos foram recuperados por vídeo-laparoscopia, caracterizados quanto à morfologia e utilizados para determinação dos estágios do ciclo meiótico por análise citogenética e maturação pela caracterização de metáfase II. Dos 33 oócitos de jaguatirica avaliados 12% (n=4) apresentaram cromossomos condensados em seu interior e dos 11 oócitos de gato-do-mato-pequeno avaliados 36% (n=4) apresentaram cromossomos condensados em seu interior, no entanto, nenhum oócito encontrava-se em metáfase II. Avaliação morfológica dos oócitos mostrou as mesmas características encontradas em outros mamíferos. Comparando os tratamentos, não houve diferença significativa (p>;0,05) no número total de estruturas ovarianas (folículos e corpos lúteos recentes) observadas em estimulações alternadas sucessivas, nas duas espécies estudadas. Também não houve diferença significativa em relação ao total de estruturas ovarianas encontradas em cada tratamento (5,7±1,2 eCG/hCG; 7,9±0,9 pFSH/pLH) para L. pardalis e (2,6±0,7 eCG/hCG; 2,0±0,5 pFSH/pLH) para L. tigrinus. Apesar das limitações deste experimento e do número reduzido de oócitos avaliados podemos sugerir que a aspiração folicular após estimulação ovariana com eCG/hCG e pFSH/pLH alternadamente resulta na recuperação de oócitos imaturos, que necessitam de cultivo em meios específicos para atingir o estágio de Metáfase II.This study represents the first assessment of oocyte's nuclear maturation by cytogenetical analysis in two endangered Brazilian felids: L. pardalis (n=5) and L. tigrinus (n=4). Females were treated 6 times with alternating eCG/hCG and pFSH/pLH protocols using an interval of 4 months between each treatment. Ovarian follicular development and oocytes recovery were performed through laparoscopy. Recovered oocytes were submitted to the morphological and cytogenetical analysis in order to evaluate the meiotic cycle and maturation (metaphase II), respectively. In 33 ocelot's oocytes observed 12% (n=4) showed condensed chromosomes and in 11 tigrinu's oocytes observed 36% (n=4) showed condensed chromosomes. However no oocytes in metaphase II were observed. Oocytes morphological evaluation showed the same characteristics were observed in others mammals. Our results suggest that L. pardalis and L. tigrinus do not show a decrease (p>;0.05) in ovarian response after repeated and alternate exposure to different gonadotropin treatments. In both L. pardalis and L. tigrinus, no differences (p>;0.05) were found regarding to the number of total ovarian structures (total follicles and CL) during successive gonadotropin treatments. There were no differences (p>;0.05) regarding to the total number of ovarian structures in L. pardalis (5.7 ± 1.2 eCG/hCG and 7.9 ± 0.9 pFSH/ pLH) or L. tigrinus (2.6 ± 0.7 eCG/hCG and 2.0 ± 0.5 pFSH/pLH). With these results we can suggest that follicular aspiration after ovarian stimulation with exogenous gonadotrophins results in immature oocytes, needing in vitro culture in specific mediums for metaphase II stage was observed

Validação de conjuntos diagnósticos comerciais para dosagem sérica de progesterona e estradiol em jaguatirica (Leopardus pardalis) e gato-do-matopequeno (Leopardus tigrinus)

Este estudo teve como objetivo validar os conjuntos diagnósticos comerciais DPC (Coat-A-Count - Diagnostic Products Corporation/USA), em fase sólida, para dosagem de progesterona e DSL (Diagnostic System Laboratories INC/USA), em fase líquida, para dosagem de estradiol, por radioimunoensaio, em soro de Leopardus pardalis (n=5) e Leopardus tigrinus (n=4),antes (15-30 dias) e após (24-28 horas) tratamento com gonadotropinas exógenas (hCG/ Novormon® e eCG/ Vetecor®; pFSH/Folltropin-V® e pLH/Lutropin-V®). A concentração sérica para Leopardus pardalis variou 0,005 a 0,151ng/ml para estradiol e 0,15-37,22ng/ml para progesterona. A concentração sérica para Leopardus tigrinus variou 0,012-0,147ng/ml para estradiol e 0,06-34,09ng/ml para progesterona. A sensibilidade mínima detectada foi 0,004 ng/ml para progesterona e 0,00014ng/ml para estradiol. Para progesterona o coeficiente intra-ensaio baixo foi 2,58% e alto 6,36%, já o coeficiente inter-ensaio baixo foi 0,67% e alto 5,55%. Para o estradiol o coeficiente intra-ensaio baixo foi 0,69% e alto 4,15%, sendo o coeficiente inter-ensaio baixo 1,40% e alto 3,00%. Paralelismo foi encontrado para progesterona e conjunto comercial DPC com r = 0,96 para Leopardus pardalis e r = 0,99 para Leopardus tigrinus. Para estradiol em conjunto comercial DSL paralelismo foi encontrado com r = 0,98 para Leopardus pardalis e com r = 0,99 para Leopardus tigrinus. Com estes resultados podemos concluir que o conjunto diagnóstico comercial DPC para dosagem de progesterona e DSL para dosagem de estradiol foram validados para utilização em soro de Leopardus pardalis e Leopardus tigrinus, podendo ser utilizado como ferramenta no manejo reprodutivo destas espécies visando à conservação.DPC commercial kit (Coat-A-Count - Diagnostic Products Corporation/USA) and DSL commercial kit (Diagnostic System Laboratories INC/USA) were used for progesterone and estradiol radioimmunoassay validation in serum of two endangered Brazilian felids: Leopardus pardalis (n=5) and Leopardus tigrinus (n=4) before (15-30 days) and after (24-28 hours) exogenous gonadotrophins treatment (hCG/ Novormon® and eCG/ Vetecor®; pFSH/Folltropin-V® and pLH/Lutropin-V®). Variation of estradiol concentration was 0.005 0.151 ng/ml and progesterone concentration was 0.15 - 37.22 ng/ml for Leopardus pardalis. Variation of estradiol concentration was 0.012-0.147 ng/ml and progesterone concentration was 0.15 - 37.22 ng/ml for Leopardus tigrinus. Minimum sensibility detected for progesterone was 0.004 ng/ml and for estradiol was 0.00014 ng/ml. Progesterone intra-assay coefficient was 2.58% and 6.36% and inter-assay was 0.67% and 5.55%. Estradiol intra-assay coefficient was 0.69% and 4.15%, and inter-assay was 1.40% and 3.00%. Parallelism was used for kits validation. Progesterone and DPC commercial kit Parallelism was r = 0.96 for Leopardus pardalis and r = 0.99 for Leopardus tigrinus. Estradiol and DSL commercial kit Parallelism was r = 0.98 for Leopardus pardalis and r = 0.99 for Leopardus tigrinus. In conclusion, these results showed that DPC commercial kit for serum progesterone dosages and DSL commercial kit for serum estradiol dosages can be used for Leopardus pardalis e Leopardus tigrinus. These findings are potentially valuable for the reproductive management and conservation of endangered felid populations

Temporal activity of rural free-ranging dogs: implications for the predator and prey species in the Brazilian Atlantic Forest

Domestic or free-ranging dogs (Canis lupus familiaris) can have deleterious effects on wildlife, acting as predators or competitors to native species. These impacts can be highly important in fragmented pristine habitats or well-preserved areas located in human dominated landscapes and where biodiversity values are usually high, such as those in southeastern Brazil. Here we explored the level of overlap or mismatch in the distributions of activity patterns of rural free-ranging dogs and potential wild prey (Didelphis aurita, Cuniculus paca; Sylvilagus brasiliensis) and a wild predator (Leopardus pardalis) in areas of Atlantic Forest in southeastern Brazil. We further explored the possible influence of the wild predator on the dog presence pattern detected in the territory analyzed. Our camera-trap data (714 camera-trap days) showed that while rural free-ranging dogs display a cathemeral activity pattern, with activity peaks at dusk and dawn, ocelot and prey species are mainly nocturnal. Moreover, we found no evidence of an effect of ocelot presence, the distance to human houses and the presence of native forests on site occupancy by dogs. The ocelot activity patterns in this study were similar to those already reported in previous studies. On the other hand, previous studies have indicated that that free-ranging dogs are often reported to be more diurnal, and it seems that the rural free-ranging dogs in our study area may have adjusted their behaviour to be more active at dawn and dusk periods. This might be to both maintain some overlap with potential prey, e.g. Sylvilagus brasiliensis, and also to avoid ocelots by being less active in periods when this predator is more active (which also coincides with peaks in activity for potential prey species). We hypothesize that the presence of ocelots might be influencing the temporal niche dimension of rural free-ranging dogs. As a sustainable management strategy, we propose conserving territories to promote the presence of medium to large predators in natural areas, in order to control free-ranging dogs and protect their vertebrate prey species

Anticorpos contra herpesvirus felino tipo 1 (FHV-1) em felinos silvestres do Brasil

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Comparison of methods fecal glucocorticoids metabolite analyses in ocelots (Leopardus pardalis) submited to hormonal treatment and video-laparoscopy

The objective of this study was to compare two different fecal metabolites extraction protocols32,33 and two commercial kits used to analyse fecal glucocorticoids (cortisol and corticosterone) in ocelots (Leopardus pardalis) submitted to hormonal treatment and video-laparoscopy procedure. Based on hormonal analyses, both extraction protocols exhibited similar performance. One commercial kit (ICN Biomedicals) had better results than another (DPC) on glucocorticoids metabolites evaluation.Dois metódos de extração de metabólitos fecais32,33 e dois conjuntos comerciais ("ImmuChem Doubly Antibody Corticosterone 125I RIA"- ICN Biomedicals e "Coat-a-count Cortisol 125I RIE" - DPC) foram utilizados na mensuração dos metabólitos de glicocorticóides (cortisol e corticosterona) em amostras de fezes de jaguatiricas (Leopardus pardalis) (n=3), antes e após tratamento hormonal e procedimentos de vídeo-laparoscopia. O objetivo desse estudo foi comparar e validar fisiologicamente os diferentes protocolos de extração e diferentes conjuntos comerciais na mensuração hormonal. Não houve diferença significativa com relação ao método de extração dos metabólitos. O conjunto comercial ICN Biomedicals provou ser melhor na detecção de metabólitos fecais de glicocorticóides em jaguatiricas

NEUTRALIZING ANTIBODIES AGAINST FELINE HERPESVIRUS TYPE 1 IN CAPTIVE WILD FELIDS OF BRAZIL

Abstract: Feline herpesvirus type 1 infection affects domestic cats, causing mainly upper respiratory tract diseases. Although this infection has been described in captive and free-ranging wild felids from Europe, Asia, North America, and Africa, no information is available on its occurrence among wild felids of Brazil. In this study, 250 serum samples of six species of Brazilian captive wild felids (Leopardus tigrinus, Leopardus wiedii, Herpailurus yaguarondi, Puma concolor, Leopardus pardalis, and Panthera onca) were examined for neutralizing antibodies to feline herpesvirus type 1. Positive sera were found in 72% of L. tigrinus samples, 15% of L. wiedii, 6% of L. pardalis, 8% of H. yaguarondi, 18% of P. concolor, and 14% of P. onca. The relatively low percentages of seropositivity and low antibody titers found among the last five species suggest that feline herpesvirus type 1 does not circulate extensively among these animals. Nevertheless, quarantine, serologic screening, and vaccination of newly introduced felids is recommended in zoos in order to prevent virus transmission and outbreaks of the disease among wild felids kept in captivity