Reproductive hormones monthly variation in free-ranging European wildcats: Lack of association with faecal marking.

Integrating ecophysiological and behavioural discoveries in conservation and management plans is essential to preserve scarce and elusive species such as the European wildcat (Felis silvestris). The purpose of this study was to characterize the monthly variation in the steroid reproductive hormone metabolite levels (oestradiol, progesterone and testosterone) in this species and to test its possible association with a monthly pattern of faecal marking. By collecting fresh faecal samples in Montes do Invernadeiro Natural Park (Galicia, Northwest Spain) each month, we obtained a total of 110 samples belonging to 25 different individuals. We conducted enzyme immunoassays which allowed us to track the annual variation in reproductive hormone excretion patterns in wildcat scats. Furthermore, we also evaluated the possible relation between the faeces used as marks and the reproductive hormone levels. We found that oestradiol and progesterone metabolite levels exhibited a distinct pattern, both increasing during the breeding months. Oestradiol metabolite larger peaks were found during March and April, whereas the highest concentration of progesterone metabolites appeared in July. On the contrary, testosterone metabolite levels did not significantly change depending on the month. Moreover, we did not find any evidence that the faecal marking behaviour pattern was associated with reproductive hormone metabolite levels. It seems that other factors related to habitat and food resources could be more important in the performance of this behaviour.

NOVEL PREGNANCY DIAGNOSIS AND ALTRENOGEST SUPPLEMENTATION LEAD TO SUCCESSFUL PREGNANCY IN A BLACK-FOOTED CAT (FELIS NIGRIPES).

This is the first time fecal prostaglandin F2α metabolite (PGFM) analysis and altrenogest were used to determine and maintain a pregnancy in a zoo-housed black-footed cat (BFC; Felis nigripes). The established pair had not produced offspring during the year since their arrival at Birmingham Zoo. Fecal samples were collected daily and analyzed by enzyme immunoassay for progestagen, estrogen metabolite, and PGFM concentrations. After a 4-mo separation, the pair was reintroduced, and breeding was reinitiated. Two pregnancies were suggested by sustained, elevated progestagen concentrations (mean >3 × baseline) but were confirmed by elevated PGFM concentrations (mean 4-25 × baseline) beginning about 1 mo after presumed or observed breeding. The first pregnancy was lost after ∼51 days, and altrenogest was administered (0.088 mg/kg/day) in order to help sustain the second pregnancy, which went to term, but the kitten did not survive. Ultrasonographic and radiographic evaluations of pregnancy were utilized during the second pregnancy to provide information on litter size.

Ongoing unraveling of a continental fauna: decline and extinction of Australian mammals since European settlement.

The highly distinctive and mostly endemic Australian land mammal fauna has suffered an extraordinary rate of extinction (>10% of the 273 endemic terrestrial species) over the last ∼200 y: in comparison, only one native land mammal from continental North America became extinct since European settlement. A further 21% of Australian endemic land mammal species are now assessed to be threatened, indicating that the rate of loss (of one to two extinctions per decade) is likely to continue. Australia's marine mammals have fared better overall, but status assessment for them is seriously impeded by lack of information. Much of the loss of Australian land mammal fauna (particularly in the vast deserts and tropical savannas) has been in areas that are remote from human population centers and recognized as relatively unmodified at global scale. In contrast to general patterns of extinction on other continents where the main cause is habitat loss, hunting, and impacts of human development, particularly in areas of high and increasing human population pressures, the loss of Australian land mammals is most likely due primarily to predation by introduced species, particularly the feral cat, Felis catus, and European red fox, Vulpes vulpes, and changed fire regimes.

EVALUATION OF THYROID HORMONES AND AS INFLUENCED BY TREATMENT WITH DESLORELIN IN PALLAS' CATS (OTOCOLOBUS (FELIS) MANUL).

Thyroid hormones regulate a variety of physiologic functions including metabolism, growth, and reproductive cycling, and these other hormones can impact the thyroid function via the hypothalamic-pituitary axis. For instance, the gonadotropin-releasing hormone agonist, deslorelin, used in nondomestic carnivores for contraception and behavioral control, down-regulates reproductive hormones through this mechanism and so may impact thyroid function. Due to clinical concerns of hypothyroidism in a bachelor group of adult male Pallas' cats (Otocolobus (Felis) manul) which also had deslorelin implants, serum samples from treated captive (n = 8) individuals, untreated captive (n = 25), and free-ranging (n = 9) individuals were analyzed for thyroid hormone concentrations. Total and free thyroxine (TT4 and FT4), total and free tri-iodothyronine (TT3 and FT3), and thyroid stimulating hormone (TSH) were measured although, due to sample volume limitations, not every hormone could be analyzed for every sample. Of these hormones, only FT4 was found statistically different between the deslorelin-treated and untreated groups. As samples were unevenly distributed across season, true comparison between seasons could not be made. The values reported for the untreated captive and free-ranging group, while representing a small sample size, can serve as a baseline assessment when evaluating the thyroid status of captive Pallas' cats.

Apparent total tract macronutrient and energy digestibility of 1- to- 3-day-old whole chicks, adult ground chicken, and extruded and canned chicken-based diets in African wildcats (Felis silvestris lybica).

Our objectives were to evaluate the composition of whole 1- to- 3-day-old chicks (Whole), ground adult chicken (Ground), chicken-based canned diet (Canned), and chicken-based extruded diet (Extruded); and evaluate apparent total tract energy and macronutrient digestibility of these diets by four captive African wildcats (Felis silvestrus lybica) utilizing a Latin Square design. We analyzed diets for macronutrient and mineral (Ca, P, K, Na, Mg, Fe, Cu, Mn, Zn, and S) composition, and screened for potentially pathogenic bacteria. Canned and Extruded diets tested negative for all microbes and met macronutrient and mineral recommendations for domestic cat foods [AAFCO (2012). Official publication. Oxford, IN: AAFCO]. Whole prey diets (Ground and Whole) met macronutrient requirements for domestic cats; however, they were below recommendations in some minerals [Mn, Cu, K, and Na; AAFCO (2012). Official publication. Oxford, IN: AAFCO], and tested positive for potentially pathogenic microorganisms (Salmonella, E. coli spp.). For all diets, apparent total tract organic matter digestibility was high (>85%). Organic matter digestibility was higher (P ≤ 0.05) for cats fed Ground (94%) compared to those fed Canned, Extruded, or Whole (87, 86, and 85%, respectively). Apparent total tract crude protein digestibility was lower than expected (i.e., <85%) for cats fed Extruded (81%) and fat digestibility was lower than expected (i.e., <90%) for cats fed Whole (82%). Cats fed whole prey items tested herein adequately maintained BW short-term; however, long-term studies are needed. These data indicate that there may be a need to monitor whole prey composition and when necessary, adjust the diet to account for potential deficiencies.

Applying embryo cryopreservation technologies to the production of domestic and black-footed cats.

Our objectives were (i) compare in vitro development of early cleavage stage domestic cat embryos after cryopreservation by minimal volume vitrification vs a standard slow, controlled-rate method, (ii) determine viability of vitrified domestic cat embryos by oviductal transfer into synchronous recipients and (iii) evaluate in vivo survival of black-footed cat (BFC, Felis nigripes) embryos after intra- and inter-species transfer. In vitro-derived (IVM/IVF) cat embryos were used to evaluate in vitro development after controlled-rate cryopreservation vs vitrification vs controls. Blastocyst development was similar in both groups of cryopreserved embryos (22-26%), but it was lower (p < 0.05) than that of fresh embryos (50%). After embryo transfer, four of eight recipients of vitrified embryos established pregnancies--three of six (50%) and one of two (50%) that received embryos from in vivo- and in vitro-matured oocytes, respectively. Three male and two female kittens weighing from 51 to 124 g (mean = 88 g) were delivered on days 61-65 of gestation. In BFC, four intra-species embryo transfer procedures were carried out--two recipients received fresh day 2 embryos (n = 5, 8) and two recipients received embryos that had been cryopreserved on day 1 (n = 6) or 2 (n = 8). A 2-year-old recipient of cryopreserved embryos established pregnancy and delivered two live male kittens. Subsequently, five cryopreserved BFC embryos were transferred to a domestic cat recipient. On day 29, the recipient was determined to be pregnant and delivered naturally a live, healthy female BFC kitten on day 66. In summary, in vivo survival of vitrified domestic cat embryos was shown by the births of kittens after transfer into recipients. Also, we demonstrated that sperm and embryo cryopreservation could be combined with intra- and inter-species embryo transfer and integrated into the array of assisted reproductive techniques used successfully for propagation of a rare and vulnerable felid species, the black-footed cat.

Scriptaid and 5-aza-2'deoxycytidine enhanced expression of pluripotent genes and in vitro developmental competence in interspecies black-footed cat cloned embryos.

Somatic cell nuclear transfer offers the possibility of preserving endangered species including the black-footed cat, which is threatened with extinction. The effectiveness and efficiency of somatic cell nuclear transfer (SCNT) depends on a variety of factors, but 'inappropriate epigenetic reprogramming of the transplanted nucleus is the primary cause of the developmental failure of cloned embryos. Abnormal epigenetic events such as DNA methylation and histone modifications during SCNT perturb the expression of imprinted and pluripotent-related genes that, consequently, may result in foetal and neonatal abnormalities. We have demonstrated that pregnancies can be established after transfer of black-footed cat cloned embryos into domestic cat recipients, but none of the implanted embryos developed to term and the foetal failure has been associated to aberrant reprogramming in cloned embryos. There is growing evidence that modifying the epigenetic pattern of the chromatin template of both donor cells and reconstructed embryos with a combination of inhibitors of histone deacetylases and DNA methyltransferases results in enhanced gene reactivation and improved in vitro and in vivo developmental competence. Epigenetic modifications of the chromatin template of black-footed cat donor cells and reconstructed embryos with epigenetic-modifying compounds enhanced in vitro development, and regulated the expression of pluripotent genes, but these epigenetic modifications did not improve in vivo developmental competence.

Use of spatial capture-recapture modeling and DNA data to estimate densities of elusive animals.

Assessment of abundance, survival, recruitment rates, and density (i.e., population assessment) is especially challenging for elusive species most in need of protection (e.g., rare carnivores). Individual identification methods, such as DNA sampling, provide ways of studying such species efficiently and noninvasively. Additionally, statistical methods that correct for undetected animals and account for locations where animals are captured are available to efficiently estimate density and other demographic parameters. We collected hair samples of European wildcat (Felis silvestris) from cheek-rub lure sticks, extracted DNA from the samples, and identified each animals' genotype. To estimate the density of wildcats, we used Bayesian inference in a spatial capture-recapture model. We used WinBUGS to fit a model that accounted for differences in detection probability among individuals and seasons and between two lure arrays. We detected 21 individual wildcats (including possible hybrids) 47 times. Wildcat density was estimated at 0.29/km² (SE 0.06), and 95% of the activity of wildcats was estimated to occur within 1.83 km from their home-range center. Lures located systematically were associated with a greater number of detections than lures placed in a cell on the basis of expert opinion. Detection probability of individual cats was greatest in late March. Our model is a generalized linear mixed model; hence, it can be easily extended, for instance, to incorporate trap- and individual-level covariates. We believe that the combined use of noninvasive sampling techniques and spatial capture-recapture models will improve population assessments, especially for rare and elusive animals.

In vitro fertilization and sperm cryopreservation in the black-footed cat (Felis nigripes) and sand cat (Felis margarita).

Studies of in vitro fertilization (IVF) and sperm cryopreservation have been conducted in several small cat species, but virtually no data exist for black-footed cats (Felis nigripes) (BFCs) or sand cats (Felis margarita) (SCs). The objectives of this study were 1) to compare in vitro motility and acrosome status of fresh and cryopreserved (frozen in pellets on dry ice or in straws in liquid nitrogen vapor) BFC and SC spermatozoa cultured in feline-optimized culture medium (FOCM) or Ham F-10, 2) to assess ovarian responsiveness in BFCs and SCs following exogenous gonadotropin treatment and laparoscopic oocyte recovery, and 3) to evaluate the fertility of fresh and frozen-thawed spermatozoa from both species using homologous and heterologous (domestic cat oocytes) IVF in the two culture media. Motility and acrosomal integrity of fresh and frozen-thawed spermatozoa from BFCs and SCs were similar (P > 0.05) in both media during 6 h of culture. Although effects were more pronounced in SCs, cryopreservation in straws was superior (P < 0.05) to cryopreservation in pellets for both species. Gonadotropin stimulation produced approximately 16 ovarian follicles per female, and >80% of recovered oocytes were of optimal (grade 1) quality. The BFC and SC spermatozoa fertilized 60.0%-79.4% of homologous and 37.7%-42.7% of heterologous oocytes in both culture media, with increased (P < 0.05) cleavage of homologous (SC) and heterologous (BFC and SC) oocytes in FOCM. These results provide the first information to date on the gamete biology of two imperiled cat species and further our capacity to apply reproductive technologies for their conservation.

Influence of feeding raw or extruded feline diets on nutrient digestibility and nitrogen metabolism of African wildcats (Felis lybica).

The African wildcat is one of the closest ancestors to the domestic cat and is believed to have similar nutrient requirements, but research is lacking. The objective of this study was to determine the effects of feeding a high-protein extruded kibble diet vs. a raw meat diet on nutrient digestibility, nitrogen metabolism, and blood metabolite concentrations in African wildcats. Five wildcats were randomized onto either a high-protein (>50% crude protein) extruded kibble diet or a raw meat diet. The study was executed as a crossover design, with 21-d periods, consisting of a 16-d adaptation phase followed by a 4-d total fecal and urine collection phase. Cats were housed individually in metabolism cages and fed to maintain body weight (BW). A fresh fecal sample was collected for short-chain fatty acid (SCFA) and branched-chain fatty acid (BCFA) analyses. Blood was analyzed for serum chemistry and leptin concentration. Food intake (as is) did not differ (P>0.05) between diets. Food intake and fecal output (g/d DMB) were greater (P<0.05) when cats consumed the kibble diet. Protein digestibility was higher (P<0.05) when cats were fed the raw meat diet vs. the kibble diet. Nitrogen intake was greater (P<0.05) when cats consumed the kibble diet, and more (P<0.05) nitrogen was present in the feces; however, the percentage of nitrogen retained and nitrogen balance did not differ (P>0.05). Fecal scores, ammonia, SCFA, and BCFA concentrations did not differ (P>0.05) between diets. Fecal butyrate molar ratio was higher (P<0.01) when cats consumed the kibble diet. Blood analyses demonstrated few differences between diets, but alanine aminotransferase activity and bicarbonate were higher (P<0.05) when cats consumed the commercial raw meat diet. Owing to lack of differences, these data indicate that African wildcats can readily utilize a high protein extruded kibble diet, and may be able to replace a commercial raw meat diet.

The nutrition and natural history of the serval (Felis serval) and caracal (Caracal caracal).

There is little information regarding the specific nutrient and dietary needs of many species of exotic cats, including those kept sporadically as house pets, such as the serval and the caracal. The diets of exotic cats kept in captivity are usually based on the nutrient requirements of the domestic cat, although there is some evidence that different cat species may not metabolize certain nutrients in the same manner as domestic species. This article provides information on the natural diet and behavior of the serval and caracal and offers insight into some health issues that may arise in a domestic environment.

Sperm evaluation of Jungle Cat (Felis chaus) obtained by urethral catheterization (CT) after medetomidine administration.

This study aimed to evaluate semen from Jungle Cat (Felis chaus) by urethral catheterization (CT) after medetomidine administration that offers feasible and different approaches to obtaining good quality sperm, especially in wild felids. Accordingly, this method was tested in five Jungle Cats. After general anesthesia with the α2-agonist medetomidine (which also stimulates semen release into the urethra) and ketamine, an abdomen ultrasound was performed to locate dilation of the first segment of the urethra (prostatic urethra). A commercial Tom cat urinary catheter 3-5 (depending on the size of the animal) was advanced into the urethra to reach the semen full dilated primary region of the urethra, so as to allow semen collection into the lumen of the catheter by capillary forces. After retraction, sperm volumes between 69 ± 27.92 yielded motility of 77.13 ± 14.15 (mean ± SD) with a mean sperm concentration of 75.13 ± 17.05 million/ml. The results of this study showed that semen collection in jungle cat is feasible, using this method. This study describes a simple, useful in field, inexpensive method which does not require the training of the animal and is better than other methods. Samples have normal pH, suitable color and consolidation, high concentration and lower contamination with excellent motility in Jungle Cat and potentially, other wild felid species, as an alternative to electro-ejaculation.

Effects of Hand-Rearing on Reproductive Success in Captive Large Cats Panthera tigris altaica, Uncia uncia, Acinonyx jubatus and Neofelis nebulosa.

Species Survival Plans and European Endangered Species Programmes have been developed for several species of endangered felids in order to build up captive reserve populations and support their conservation in the wild. The Siberian tiger (Panthera tigris altaica), snow leopard (Uncia uncia), cheetah (Acinonyx jubatus) and clouded leopard (Neofelis nebulosa) are managed in such ex situ conservation programmes. Many zoological institutions hand-rear offspring if rearing by the mother fails. Hand-rearing can cause behavioural problems, resulting in decreased copulation and lower breeding success in some species. In this study, studbook data subsets were examined: from 1901 to 2011; and 2000 to 2011. We analysed records from 4273 Siberian tigers, 2045 snow leopards, 3435 cheetahs, and 804 clouded leopards. We assessed the number of offspring produced, litter size, age at first reproduction, longevity, infant mortality and generational rearing of hand-reared versus parent-reared individuals. Hand-reared Siberian tigers (p<0.01; p = 0.0113), snow leopards (p<0.01), male cheetahs (p<0.01) and female clouded leopards (p<0.01) produced fewer offspring than parent-reared individuals. Hand-reared snow leopard breeding pairs had larger litters than parent-reared pairs (p = 0.0404). Hand-reared snow leopard females reproduced later in life (p<0.01). Hand-reared female Siberian tigers lived shorter lives, while hand-reared cheetahs lived longer (p<0.01; p = 0.0107). Infant mortality was higher in hand-reared snow leopards (p<0.01) and male cheetahs (p = 0.0395) in the 1901-2011 dataset and lower in hand-reared female Siberian tiger and male snow leopard cubs (p = 0.0404; p = 0.0349) in the 2000-2011 dataset. The rearing of the mother and subsequent rearing of offspring showed a significant relationship for all species (p<0.01 for Siberian tiger and snow leopard cubs; p<0.001 for cheetah and snow leopard cubs). Taking into account the limited carrying capacity of zoos, the results of this study highlight that careful consideration should be taken when deciding whether or not to hand-rear individuals that are part of Species Survival Plans and European Endangered Species Programmes.

[African wild cat (Felis lybica Forster, 1780) in the Saratov Transvolga Region].

Analysis of the area limits where African wild cat was found as well as of published data on its distribution in the Saratov Transvolga Region demonstrated that the local range of this species extended and its northwestern limit advanced almost 1000 km to the Volga zoogeographical boundary.

Domestic Cats (Felis silvestris catus) Do Not Show Signs of Secure Attachment to Their Owners.

The Ainsworth Strange Situation Test (SST) has been widely used to demonstrate that the bond between both children and dogs to their primary carer typically meets the requirements of a secure attachment (i.e. the carer being perceived as a focus of safety and security in otherwise threatening environments), and has been adapted for cats with a similar claim made. However methodological problems in this latter research make the claim that the cat-owner bond is typically a secure attachment, operationally definable by its behaviour in the SST, questionable. We therefore developed an adapted version of the SST with the necessary methodological controls which include a full counterbalance of the procedure. A cross-over design experiment with 20 cat-owner pairs (10 each undertaking one of the two versions of the SST first) and continuous focal sampling was used to record the duration of a range of behavioural states expressed by the cats that might be useful for assessing secure attachment. Since data were not normally distributed, non-parametric analyses were used on those behaviours shown to be reliable across the two versions of the test (which excluded much cat behaviour). Although cats vocalised more when the owner rather the stranger left the cat with the other individual, there was no other evidence consistent with the interpretation of the bond between a cat and its owner meeting the requirements of a secure attachment. These results are consistent with the view that adult cats are typically quite autonomous, even in their social relationships, and not necessarily dependent on others to provide a sense of security and safety. It is concluded that alternative methods need to be developed to characterise the normal psychological features of the cat-owner bond.

Lasting secondary antiepileptogenesis induced by cingulate kindling.

PURPOSE: To test the validity of a hypothesis that kindling-induced enduring kindling inhibition of the homotopic secondary site is not a primate species-specific effect but occurs also in feline species. METHODS: Five mature male cats were kindled at the anterior (AC) and posterior (PC) cingulate cortex followed by secondary-site kindling and primary-site retest. RESULTS: Kindling was characterized by a marked seizure-stage instability and a progressive afterdischarge threshold elevation in all the animals. A protracted nonconvulsive seizure stage was followed by convulsive evolution and rapid generalization. At the secondary site, positive transfer effect was absent, and kindling did not occur. Primary-site retest activated kindled seizure within three stimulations. Recurrent spontaneous seizures, identical to the primary-site kindled seizure, occurred in two AC-kindled animals. Subsequent to this, a markedly elevated afterdischarge threshold also was found at some distant cortical and subcortical sites, to which the afterdischarge propagated. Kindling of the amygdala ipsilateral to the kindled AC or PC was tardy, with no positive transfer effect. However, it was associated with afterdischage threshold reduction. CONCLUSIONS: Cingulate kindling-induced enduring kindling inhibition at a homotopic secondary site is not a primate species-specific effect and occurs also in feline species. It is not limited to a homotopic site and represents a lasting secondary antiepileptogenesis, presumably due to enhanced intrinsic inhibitory mechanism(s) available to the mammalian brain. A selective afterdischarge threshold elevation concurrently observed is suspected to be a feline species-specific effect.

Avaliação de alguns parâmetros ecocardiográficos do gato-do-mato (Leopardus tigrinus), mantido em cativeiro e submetido à anestesia com xilazina e quetamina / Evaluation of some echocardiographic parameters of Oncilla (L. tigrinus), kept in captivity and submitted to anesthesia with xilazine and ketamine

Avaliaram-se alguns parâmetros ecocardiográficos em modos B, M e Doppler de 27 gatos-do-mato, Leopardus tigrinus, pequeno felídeo selvagem, mantidos em cativeiro e submetidos à anestesia com 1 a 2mg/kg de xilazina e 10mg/kg de quetamina. Observaram-se alterações dos parâmetros cardiovasculares quando os resultados foram comparados aos do gato doméstico (Felis catus) não anestesiado.

Some echocardiographic parameters in B, M-mode and Doppler of 27 Oncillas, Leopardus tigrinus, a wild little feline, kept in captivity and submitted to anesthesia with 1 to 2mg/kg of xilazine and 10mg/kg of ketamine, had been evaluated. Changes of the cardiovascular parameters were observed when the results were compared to non anesthetized domestic cat (Felis catus).