Colheita farmacológica de sêmen de onças-pardas (Puma concolor: Mammalia: Carnivora: Felidae) / Pharmacological semen collection in cougars (Puma concolor: Mammalia: Carnivora: Felidae)

Objetivou-se, por meio do presente estudo, avaliar o método de colheita farmacológica de sêmen com sondagem uretral, em machos de onças-pardas (Puma concolor) mantidos em cativeiro. A técnica proposta (Cat; N=3) foi comparada com a eletroejaculação (EE; N=4). Para a colheita farmacológica, utilizou-se medetomidina para induzir a liberação de sêmen na uretra e sonda uretral para gatos, sem janela lateral, para colheita do sêmen por capilaridade. O método foi eficaz em todos os animais usados. Por meio dessa técnica, colheram-se amostras com menor volume (106,7±30,5aµL) e maior concentração (524,1±54,3b x 106 espermatozoides/mL) em relação à EE (450,0±0,1bµL e 205,0±141,8a x 106 espermatozoides/mL). As avaliações de vigor, motilidade e patologia espermática demonstraram que a técnica não afeta a qualidade do sêmen em relação à EE (P>0,05). Dessa forma, o método proposto consiste em uma técnica mais prática e eficiente para a colheita de sêmen com boa qualidade, dispensando o eletroejaculador.(AU)

The aim of this study was to evaluate the pharmacological semen collection method with urethral catheterization (CT) in captive cougar (Puma concolor) males. The pharmacological method (CT; N= 3) was compared to the electroejaculation technique (EE; N= 4). For CT collection, medetomidine was administrated to induce semen release using a tomcat catheter inserted into the urethra to collect by capillarity. The proposed method was efficacious on all animals used. Through the CT method, semen collected yielded smaller volume (106,7±30,5aµL) and higher concentration (524,1±54,3b x 106sperm/mL) compared to EE (450,0±0,1bµL and 205,0±141,8a x 106 sperm /mL). Evaluations of vigor, motility and sperm pathology demonstrated that CT does not affect semen quality when compared to EE (P> 0.05). Thus, the proposed method consists of a more practical and efficient technique for semen collection with good quality, eliminating the need for eletroejaculation.(AU)

The effect of habitat fragmentation on the genetic structure of a top predator: loss of diversity and high differentiation among remnant populations of Atlantic Forest jaguars (Panthera onca).

Habitat fragmentation may disrupt original patterns of gene flow and lead to drift-induced differentiation among local population units. Top predators such as the jaguar may be particularly susceptible to this effect, given their low population densities, leading to small effective sizes in local fragments. On the other hand, the jaguar's high dispersal capabilities and relatively long generation time might counteract this process, slowing the effect of drift on local populations over the time frame of decades or centuries. In this study, we have addressed this issue by investigating the genetic structure of jaguars in a recently fragmented Atlantic Forest region, aiming to test whether loss of diversity and differentiation among local populations are detectable, and whether they can be attributed to the recent effect of drift. We used 13 microsatellite loci to characterize the genetic diversity present in four remnant populations, and observed marked differentiation among them, with evidence of recent allelic loss in local areas. Although some migrant and admixed individuals were identified, our results indicate that recent large-scale habitat removal and fragmentation among these areas has been sufficiently strong to promote differentiation induced by drift and loss of alleles at each site. Low estimated effective sizes supported the inference that genetic drift could have caused this effect within a short time frame. These results indicate that jaguars' ability to effectively disperse across the human-dominated landscapes that separate the fragments is currently very limited, and that each fragment contains a small, isolated population that is already suffering from the effects of genetic drift.

Hematology and blood chemistry parameters differ in free-ranging maned wolves (Chrysocyon brachyurus) living in the Serra da Canastra National Park versus adjacent farmlands, Brazil.

There has been growing interest in the specific impacts of anthropogenic factors on the health of wildlife. This study examined hematology and serum chemistry status of a prominent carnivore, the maned wolf (Chrysocyon brachyurus), living in, on the boundaries to, or on adjacent farmlands to the Serra da Canastra National Park, Brazil. Twenty-eighty wolves were captured, and values were compared 1) between subadults (n=8 animals) and adults (n=20 animals), 2) males (n=12 animals) and females (n=16 animals), and 3) among wolves living inside the park (n=11), near the park border (n=11 animals), and in neighboring farming areas (n=6 animals). Age, gender, and wolf locations influenced (P<0.05) hematology and serum biochemistry values. Specifically, adults had lower (P<0.05) circulating phosphorus than subadults. Males had lower (P<0.05) serum glucose, creatinine phosphokinase, and cholesterol and higher (P<0.05) potassium than females. Erythrocyte count and serum cholinesterase were lower (P<0.05) in wolves living within the park compared with near the park border or on farmlands. Mean corpuscular volume was lower (P<0.05) in wolves living near the park border than those ranging within the park and on farmlands. Aspartate transaminase and chloride were higher (P<0.05) in wolves living inside the park compared with those ranging near the park border. Creatinine phosphokinase was lower (P<0.05) in wolves living on farmland compared with the other two locations. These results clearly reveal a relationship between age and gender on hematology and serum biochemistry values in free-living maned wolves. More importantly, certain traits indicative of health are potentially compromised in wolves living in areas under anthropogenic pressure. These data lay a foundation for examining the influence of farming and local domestic species on disease susceptibility and fitness in the maned wolf.

Duration of spermatogenesis and daily sperm production in the jaguar (Panthera onca).

The jaguar, like most wild felids, is an endangered species. Since there are few data regarding reproductive biology for this species, our main goal was to investigate basic aspects of the testis and spermatogenesis. Four adult male jaguars were utilized; to determine the duration of spermatogenesis, two animals received an intratesticular injection of H(3)-thymidine. Mean (+/-SEM) testis weight and the gonadosomatic index were 17.7+/-2.2g and 0.05+/-0.01%, respectively, whereas the seminiferous tubules and the Leydig cells volume density were 74.7+/-3.8 and 16.7+/-1.6%. Eight stages of spermatogenesis were characterized, according to the tubular morphology system and acrosome development. Each spermatogenic cycle and the entire spermatogenic process (based on 4.5 cycles) lasted approximately 12.8+/-0.01 and 57.7+/-0.07 d. The number of Sertoli and Leydig cells per gram of testis was 29+/-4 x 10(6) and 107+/-12 x 10(6). Based on the number of round spermatids per pachytene spermatocyte (2.8+/-0.3:1; meiotic index); significant cell loss (30%) occurred during the two meiotic divisions. There were approximately eight spermatids for each Sertoli cell (Sertoli cell efficiency), whereas the daily sperm production per gram of testis was 16.9+/-1.2 x 10(6). We expect that in the near future, the knowledge obtained in the present investigation will facilitate, utilizing germ cell transplantation, preservation of the germinal epithelium and the ability to generate sperm from jaguars in testes of domestic cats.

Comparison of serum hormone levels of captive and free-living maned wolves Chrysocyon brachyurus.

Serum hormone levels were compared between captive and free-living maned wolves and seasonal variations of sex hormones were studied. Blood samples were collected from 16 male and 26 female adult animals from Brazilian zoos, and from 30 male and 24 female free-living adults to determine serum progesterone and testosterone by radioimmunoassay. Serum testosterone concentrations varied (P < 0.05) across seasons for 16 captive males, being higher in autumn (2184.7 +/- 355.1 pg/mL) than in summer (1080.7 +/- 205.4 pg/mL), winter (1270.1 +/- 276.6 pg/mL) and spring (963.9 +/- 248.1 pg/mL), although they did not differ between summer, winter and spring. Testosterone concentration of 30 free-living males differed (P < 0.05) between autumn (824.1 +/- 512.2 pg/mL), winter (14.4 +/- 8.0 pg/mL) and spring (151.9 +/- 90.5 pg/mL). Comparison between captive and free-living animals showed no difference in autumn (P > 0.05). Sixteen captive males showed higher testosterone concentration during winter and spring compared with 30 free-living animals (P < 0.05). Progesterone concentration varied among seasons in 26 captive females (P < 0.05), being higher in autumn (15.3 +/- 3.1 ng/mL) than in summer (6.6 +/- 1.5 ng/mL), winter (5.3 +/- 3.1 ng/mL) and spring (4.3 +/- 0.7 ng/mL). Progesterone concentration of 24 free-living females varied between autumn (17.1 +/- 6.0 ng/mL) and winter (1.7 +/- 0.3 ng/mL) (P < 0.05), but we could not obtain data for spring or summer. No difference in progesterone levels was observed between captive and free-living females in autumn and winter.

Comparison of serum hormone levels of captive and free-living maned wolves Chrysocyon brachyurus

Serum hormone levels were compared between captive and free-living maned wolves and seasonal variations of sex hormones were studied. Blood samples were collected from 16 male and 26 female adult animals from Brazilian zoos, and from 30 male and 24 female free-living adults to determine serum progesterone and testosterone by radioimmunoassay. Serum testosterone concentrations varied (P < 0.05) across seasons for 16 captive males, being higher in autumn (2184.7 ± 355.1 pg/mL) than in summer (1080.7 ± 205.4 pg/mL), winter (1270.1 ± 276.6 pg/mL) and spring (963.9 ± 248.1 pg/mL), although they did not differ between summer, winter and spring. Testosterone concentration of 30 free-living males differed (P < 0.05) between autumn (824.1 ± 512.2 pg/mL), winter (14.4 ± 8.0 pg/mL) and spring (151.9 ± 90.5 pg/mL). Comparison between captive and free-living animals showed no difference in autumn (P > 0.05). Sixteen captive males showed higher testosterone concentration during winter and spring compared with 30 free-living animals (P < 0.05). Progesterone concentration varied among seasons in 26 captive females (P < 0.05), being higher in autumn (15.3 ± 3.1 ng/mL) than in summer (6.6 ± 1.5 ng/mL), winter (5.3 ± 3.1 ng/mL) and spring (4.3 ± 0.7 ng/mL). Progesterone concentration of 24 free-living females varied between autumn (17.1 ± 6.0 ng/mL) and winter (1.7 ± 0.3 ng/mL) (P < 0.05), but we could not obtain data for spring or summer. No difference in progesterone levels was observed between captive and free-living females in autumn and winter.

Changes in the fecal concentrations of cortisol and androgen metabolites in captive male jaguars (Panthera onca) in response to stress

In the present study we determined the efficacy of the measurement of fecal cortisol and androgen metabolite concentrations to monitor adrenal and testicular activity in the jaguar (Panthera onca). Three captive male jaguars were chemically restrained and electroejaculated once or twice within a period of two months. Fecal samples were collected daily for 5 days before and 5 days after the procedure and stored at -20ºC until extraction. Variations in the concentrations of cortisol and androgen metabolites before and after the procedure were determined by solid phase cortisol and testosterone radioimmunoassay and feces dry weight was determined by drying at 37ºC for 24 h under vacuum. On four occasions, fecal cortisol metabolite levels were elevated above baseline (307.8 ± 17.5 ng/g dry feces) in the first fecal sample collected after the procedure (100 to 350 percent above baseline). On one occasion, we did not detect any variation. Mean (± SEM) fecal androgen concentration did not change after chemical restraint and electroejaculation (before: 131.1 ± 26.7, after: 213.7 ± 43.6 ng/g dry feces). These data show that determination of fecal cortisol and androgen metabolites can be very useful for a noninvasive assessment of animal well-being and as a complement to behavioral, physiological, and pathological studies. It can also be useful for the study of the relationship between adrenal activity and reproductive performance in the jaguar.

Changes in the fecal concentrations of cortisol and androgen metabolites in captive male jaguars (Panthera onca) in response to stress.

In the present study we determined the efficacy of the measurement of fecal cortisol and androgen metabolite concentrations to monitor adrenal and testicular activity in the jaguar (Panthera onca). Three captive male jaguars were chemically restrained and electroejaculated once or twice within a period of two months. Fecal samples were collected daily for 5 days before and 5 days after the procedure and stored at -20 degrees C until extraction. Variations in the concentrations of cortisol and androgen metabolites before and after the procedure were determined by solid phase cortisol and testosterone radioimmunoassay and feces dry weight was determined by drying at 37 degrees C for 24 h under vacuum. On four occasions, fecal cortisol metabolite levels were elevated above baseline (307.8 +/- 17.5 ng/g dry feces) in the first fecal sample collected after the procedure (100 to 350% above baseline). On one occasion, we did not detect any variation. Mean (+/- SEM) fecal androgen concentration did not change after chemical restraint and electroejaculation (before: 131.1 +/- 26.7, after: 213.7 +/- 43.6 ng/g dry feces). These data show that determination of fecal cortisol and androgen metabolites can be very useful for a noninvasive assessment of animal well-being and as a complement to behavioral, physiological, and pathological studies. It can also be useful for the study of the relationship between adrenal activity and reproductive performance in the jaguar.

Comparative analyses of semen and endocrine characteristics of free-living versus captive jaguars (Panthera onca).

Semen and blood samples were obtained from free-living (n = 6) and captive (n = 8) jaguars (Panthera onca) to compare reproductive characteristics between the two populations. Semen samples were analysed for volume (ml), percentage of motile spermatozoa, rate of forward progression (0-5), concentration (10(6) ml(-1)), total sperm count (10(6)) and sperm morphology. Serum testosterone concentration was determined by radioimmunoassay. Although ejaculate volume was greater in captive jaguars (n = 47 samples) than in free-living jaguars (n = 7 samples) (P < 0.05), the free-living jaguars produced more total spermatozoa (59.3 +/- 12.8 versus 152.0 +/- 88.0 x 10(6), respectively; not significant) with better viability and forward progression (2.8 +/- 0.1 versus 3.5 +/- 0.2, respectively; P < 0.05) and more spermatozoa with normal morphology (73.5 +/- 3.9 versus 5.0 +/- 1.1%, respectively; P < 0.05). Serum testosterone concentrations were similar for captive and free-living male jaguars (3.1 +/- 0.7 and 2.1 +/- 0.8 ng ml(-1), respectively). In summary, the data showed that semen may be collected successfully from free-living jaguars and evaluated under field conditions to establish normative reproductive values in this species. The results also indicate that jaguars maintained in zoos show inferior seminal characteristics compared with free-living animals.

Colheita e avaliaçäo do sêmen em onça pintada (Panthera onca) / Semen collection and evaluation in the jaguar (Panthera onca)

Foram submetidas à eletroejaculaçäo 10 onças pintadas, de origem desconhecida, mantidas na Fundaçäo Parque Zoológico de Säo Paulo (n = 7) e Parque Zoológico Municipal "Quinzinho de Barros"- Sorocaba (n = 3), recebendo alimentaçäo de acordo com a dieta de rotina de cada instituiçäo. Nos 54 procedimentos realizados, foram obtidos ejaculados, ocorrendo contaminaçäo por urina em 3,7 por cento (n = 2) e em 11 (20,4 por cento) ocasiöes em que näo foram observados espematozóides. O volume médio obtido foi 7,42 ñ 3,69 ml, com motilidade média de 62,6 ñ 11,0 por cento e vigor de 2,71 ñ 0,52. A concentraçäo espermática foi 6,20 ñ 3,03 x 10 elevado a sexta potência espematozóides/ml, sendo que a porcentagem média de espermatozóides morfologicamente normais foi 46,7 ñ 5,8 por cento. A técnica de colheita de sêmen por eletroejaculaçäo é eficiente na espécie Panthera onça