María Delclaux

POST-THAW VIABILITY OF EUROPEAN BISON (BISON BONASUS) SEMEN FROZEN WITH EXTENDERS CONTAINING EGG YOLK OR LIPIDS OF PLANT ORIGIN AND EXAMINED WITH A HETEROLOGOUS IN VITRO FERTILIZATION ASSAY

Abstract Basic characteristics of European bison (Bison bonasus) semen were described and the efficacies of two extenders—Triladyl, containing egg yolk, and a synthetic extender, containing soybean lipids—were tested for semen cryopreservation. Seven ejaculates were collected by electroejaculation from a 10-yr-old, European bison bull. Each ejaculate was diluted at 37°C to a final concentration of 200 × 106 sperm/ml with Triladyl or the synthetic extender. Extended semen samples were frozen according to a standard bull semen freezing protocol. After 2 wk of storage, one straw from each extender and ejaculate was thawed, and postthaw quality was evaluated by individual sperm motility and movement rate, numbers of sperm morphologic abnormalities and intact acrosomes, functional integrity of the sperm membranes determined by hypoosmotic swelling test (HOST), viability (live–dead, eosin–nigrosin stain), and a heterologous in vitro sperm penetration assay (SPA). A total of 600 in vitro–matured bovine oocytes were inseminated with 1 × 106 spermatozoa of Holstein semen frozen–thawed in Triladyl (control) or of European bison semen frozen in Triladyl or the synthetic extender. Nuclear status of the oocytes was determined after 18 h of sperm–oocyte coincubation. Extender had no effect on any evaluated parameters of semen after dilution and cooling (4 hr at 5°C) or in postthaw individual motility, quality of movement, and sperm morphology. However, significantly (P < 0.05) higher numbers of spermatozoa with intact acrosomes, intact membranes (HOST), and viable sperm (P < 0.01) were in semen frozen in Triladyl than in the synthetic extender. Mean values for heterologous SPA for bull (control) and for bison semen frozen in the synthetic extender were very much alike—63.3 ± 10.6% and 63.1 ± 15.9%, respectively; bison semen frozen in Triladyl was lower, 43.0 ± 24.2% but not significantly different. Cumulative results from a variety of viability assays of diluted/cooled and frozen–thawed semen, including the heterologous SPA, suggest that European bison semen can be successfully frozen in both extenders tested in this study.

Comprehensive serological analysis of two successive heterologous vaccines against H5N1 avian influenza virus in exotic birds in zoos.

ABSTRACT In 2005, European Commission directive 2005/744/EC allowed controlled vaccination against avian influenza (AI) virus of valuable avian species housed in zoos. In 2006, 15 Spanish zoos and wildlife centers began a vaccination program with a commercial inactivated H5N9 vaccine. Between November 2007 and May 2008, birds from 10 of these centers were vaccinated again with a commercial inactivated H5N3 vaccine. During these campaigns, pre- and postvaccination samples from different bird orders were taken to study the response against AI virus H5 vaccines. Sera prior to vaccinations with both vaccines were examined for the presence of total antibodies against influenza A nucleoprotein (NP) by a commercial competitive enzyme-linked immunosorbent assay (cELISA). Humoral responses to vaccination were evaluated using a hemagglutination inhibition (HI) assay. In some taxonomic orders, both vaccines elicited comparatively high titers of HI antibodies against H5. Interestingly, some orders, such as Psittaciformes, which did not develop HI antibodies to either vaccine formulation when used alone, triggered notable HI antibody production, albeit in low HI titers, when primed with H5N9 and during subsequent boosting with the H5N3 vaccine. Vaccination with successive heterologous vaccines may represent the best alternative to widely protect valuable and/or endangered bird species against highly pathogenic AI virus infection.

VALIDATING A COMMERCIALLY AVAILABLE ENZYME IMMUNOASSAY FOR THE DETERMINATION OF 17β-ESTRADIOL AND PROGESTOGENS IN THE FECES OF CHEETAHS (ACINONYX JUBATUS): A CASE REPORT

Abstract Fecal 17β-estradiol and progestogens excretion was monitored in adult, female cheetahs (Acinonyx jubatus; n = 2), ZGG-12301 (born 3 April 1993), gonadotrophin treated and ZGT-3301, (born 19 August 1993), nontreated, for 120 days using commercially available plate enzyme immunoassay kits prepared for human serum or plasma. There were significant differences (P < 0.001) between baseline and peak concentrations of both hormone measures. Female ZGG-12301, which conceived, but this pregnancy resulted in an unobserved spontaneous abortion, showed no significant difference (P > 0.05) between baseline and gestation 17β-estradiol values; fecal 17β-estradiol excretion during pregnancy was statistically different (P < 0.001) from excretion during the nonpregnancy period. Baseline progestogen concentrations were different from pregnancy (P < 0.001) and postovulatory (P < 0.01) concentrations, and progestogen concentrations during pregnancy period were different (P < 0.001) from postovulatory concentrations. In the nontreated cheetah (ZGT-3301), basal and increased progestogen concentrations were statistically different (P < 0.01). On the basis of 17β-estradiol excretory patterns, duration of the estrous cycle (x̄ ± SEM) was 13.2 ± 2.2 days. These results suggest that the enzyme-linked immunosorbent methods reported in this study were capable of quantifying reproductive hormones in fecal extracts of cheetahs and could be a practical alternative to other enzyme-linked immunosorbent assays which require more complex procedures.

Aggressive behavior and hair cortisol levels in captive Dorcas gazelles (Gazella dorcas) as animal-based welfare indicators

Ensuring welfare in captive wild animal populations is important not only for ethical and legal reasons, but also to maintain healthy individuals and populations. An increased level of social behaviors such as aggression can reduce welfare by causing physical damage and chronic stress to animals. Recently, cortisol in hair has been advanced as a non-invasive indicator to quantify long-lasting stress in many species. The sensitivity of social behavior and hair cortisol concentration was evaluated in several groups of dorcas gazelles (Gazella dorcas). Four different groups of gazelles from three different zoos were observed and the expression of intra-specific affiliative and negative social behaviors was assessed across the different groups. Hair samples were taken from sub-groups of animals and analyzed for cortisol concentrations. Significant differences between groups of dorcas gazelles were found in frequency of negative social behavior and hair cortisol concentration. Despite the low sample size, these two parameters had a positive Spearman correlation coefficient (rs  = +0.80, P = 0.20). These results suggest that hair cortisol levels are sensitive to differences in the social structure of dorcas gazelles. Zoo Biol. 35:467-473, 2016.

AVASCULAR NECROSIS OF THE FEMORAL HEADS IN A RED PANDA (AILURUS FULGENS FULGENS): POSSIBLE LEGG–CALVE–PERTHES DISEASE

Abstract A 17-mo-old captive-born female red panda (Ailurus fulgens fulgens) presented with a sudden onset of lameness in its left hind leg was diagnosed radiographically as having possible severe, bilateral Legg–Calve–Perthes disease with fracture of the great trochanter of the left femur. Surgical repair of the fracture was performed using pins and a tension band wire through a lateral approach to the hip. This is the first case reported at Madrid Zoo-Aquarium, where 63 individuals have been bred over 15 yr.

Fecal Steroid Evaluation to Monitor Reproductive Status in Wild Ungulate Females Using Enzyme Immunoassay Commercial Kits

Abstract: Analysis of reproductive hormones in fecal samples is necessary for the noninvasive monitoring of reproductive status in free-ranging species. The aim of the present study was to establish an easy noninvasive method to monitor reproductive status in wild ungulate females. Feces were collected daily, weekly, or three or four times a week directly from the soil for a period ranging from 1 to 9.8 mo. Fecal estradiol and progestagens were monitored in nine wild ungulate females (Barbary sheep, Ammotragus lervia [n = 3]; European bison, Bison bonasus [n = 1]; auroch, Bos taurus primigenius [n = 2]; sitatunga, Tragelaphus spekii gratus [n = 2]; and Indian rhinoceros, Rhinoceros unicornis [n = 1]) by using commercially available enzyme immunoassay kits prepared for human serum or plasma. In the species evaluated in this study, luteal phase, abortion, and gestation patterns corresponded closely with changes in fecal progestagens. Luteal phase and gestation values differed significantly (P < 0.001) from basal values, whereas progestagens values after abortion were not significantly different (P > 0.05) from basal values. For estradiol excretory patterns, follicular phase and pregnancy values differed significantly (P < 0.001) from basal values, but differences between values after abortion and basal values were not significant (P > 0.05); length of estrous cycles were clearly defined through estradiol data. This study demonstrates that ovarian function in the wild ungulate females studied can be monitored by enzyme-linked immunosorbent assay (ELISA). Therefore, ELISA methodologies used here could be a practical alternative to other ELISAs that require more complex procedures or whose commercial availability is difficult.

Using Farm Animal Welfare Protocols as a Base to Assess the Welfare of Wild Animals in Captivity—Case Study: Dorcas Gazelles (Gazella dorcas)

Simple Summary Animal welfare is gaining attention regarding the conservation of species not only due to ethical and legal reasons but also because optimal welfare can ensure stable and healthy populations. Currently, there is a lack of protocols that help to objectively assess welfare of wild animals in captivity. In this study, we have developed and applied a protocol for the assessment of welfare in captive dorcas gazelles (Gazella dorcas). We have gathered information from existing literature about the biology of this species in wild conditions, as well as in zoo husbandry, breeding, management and care guidelines developed for this species. We took a protocol developed for the on-farm welfare assessment in cattle as a reference and suggested 23 indicators that we considered useful to assess welfare in captive dorcas gazelles. To test the utility of this protocol, we then applied it in five groups of dorcas gazelles from three different zoos and we detected areas for improvement in all of the groups assessed. Abstract There is a lack of protocols specifically developed for the assessment of welfare of wild animals in captivity, even when it is known that providing good standards of welfare is important. The aim of this study was the development and the application of a protocol for the assessment of welfare in captive dorcas gazelles. The protocol was mainly developed taking into account the protocol for the assessment of welfare in cattle from the Welfare Quality® project, the available literature of the biology of this species and the Husbandry Guidelines developed for captive breeding and management of this species. The protocol was specifically developed for dorcas gazelles and included four principles, 10 criteria and 23 animal and environmental-based indicators. To test its utility, this protocol was applied to five different groups of gazelles from three different zoos. Its application made possible to detect areas for improvement in all groups assessed.