Sexual maturation in the Sumatran rhinoceros (Dicerorhinus sumatrensis).

To help save the Sumatran rhino from extinction, the captive breeding program must capitalize on each rhino's reproductive lifespan. Doing so requires knowing when calves are sexually mature. The goal of this study was to monitor physiological changes associated with sexual maturation in two captive born calves (one male and one female) to determine the approximate age of maturity for both sexes of this species.

Laparoscopic oviductal artificial insemination improves pregnancy success in exogenous gonadotropin-treated domestic cats as a model for endangered felids.

Artificial insemination (AI) in cats traditionally uses equine chorionic gonadotropin (eCG) and human chorionic gonadotropin (hCG) to induce follicular development and ovulation, with subsequent bilateral laparoscopic intrauterine insemination. However, long-acting hCG generates undesirable secondary ovulations in cats. Uterine AI also requires relatively high numbers of spermatozoa for fertilization (~8 × 10(6) sperm), and unfortunately, sperm recovery from felids is frequently poor.

Comparative fertility of freshly collected vs frozen-thawed semen with laparoscopic oviductal artificial insemination in domestic cats.

Artificial insemination (AI) is potentially invaluable as an adjunct to natural breeding for the conservation management of non-domestic felid populations. The efficacy of AI, however, must be substantially improved for applied use, especially when using frozen semen. Our recent advances in using laparoscopic oviductal AI (LO-AI) with low sperm numbers and freezing of cat semen in a soy lecithin-based cryoprotectant medium suggest that combining these two approaches might improve pregnancy outcomes with frozen-thawed spermatozoa.

Factors influencing annual fecal testosterone metabolite profiles in captive male polar bears (Ursus maritimus).

The objectives of this study were to assess the effects of season, breeding activity, age and latitude on fecal testosterone metabolite concentrations in captive, adult male polar bears (Ursus maritimus). Fourteen polar bears from 13 North American zoos were monitored for 12-36 months, producing 25-year-long testosterone profiles. Results indicated that testosterone was significantly higher during the breeding season (early January through the end of May) compared with the non-breeding season with the highest concentrations excreted from early January through late March.

Laparoscopic oviductal embryo transfer and artificial insemination in felids--challenges, strategies and successes.

Embryo transfer (ET) and artificial insemination (AI) are potentially invaluable techniques for the propagation and management of genetically valuable domestic cat and endangered nondomestic cat populations. Many of the challenges that impair the effective application of ET and AI in felids may be overcome by using laparoscopic oviductal (LO) approaches. LO-ET and LO-AI are minimally-invasive procedures, requiring only two small skin incisions for insertion of a laparoscope and grasping forceps into the abdominal cavity to permit visualization and catheterization of the oviduct for embryo or semen deposition.

Physiological effects of tourism and associated food provisioning in an endangered iguana.

Deliberately feeding wildlife is an increasingly popular tourism-related activity despite a limited understanding of long-term impacts on the species being fed. As a result, tourist behaviours that may have adverse impacts on imperiled species have often been encouraged without the necessary evaluation or oversight. Here, we report the responses of Northern Bahamian Rock Iguanas (Cyclura cychlura) to human-visitation pressure and associated food provisioning.

Improved cryopreservation of domestic cat sperm in a chemically defined medium.

The objective was to compare a proprietary egg yolk-based cryopreservation medium with a chemically defined soy-based medium, as well as to examine effects of temperature of glycerol addition on sperm parameters and IVF after freezing and thawing of domestic cat sperm. Semen was collected from adult cats (four males and three ejaculates per male), divided in four equal aliquots, and extended in either egg yolk with 4% glycerol added before (EYG) or after (EY) cooling to 5 °C, or soy-lecithin with 4% glycerol added before (SLG) or after (SL) cooling to 5 °C. Extended sperm were frozen in straws over liquid nitrogen vapor.

Longitudinal fecal hormone analysis for monitoring reproductive activity in the female polar bear (Ursus maritimus).

The objective was to identify suitable enzyme immunoassays to monitor gonadal and placental function in the female polar bear. Immunoreactive progesterone, progesterone metabolite (PdG), estrogen, and androgen metabolite (T) concentrations were measured in fecal samples collected over 24 mo from captive female bears (N = 20). Whereas fecal extracts produced displacement curves parallel to the standard curve for each respective steroid, T and PdG more accurately reflected reproductive events.

Neonatal milestones, behavior and growth rate of Sumatran rhinoceros (Dicerorhinus sumatrensis) calves born and bred in captivity.

The birth of Sumatran rhino calves at the Cincinnati Zoo and Botanical Garden offered a unique opportunity to study early development and cow-calf behavior in this elusive, critically endangered species. Study goals were to: (1) record developmental milestones of newborn calves; (2) characterize cow and calf behavior during the first year following birth; (3) compare trends in the behavioral development of a male vs. female calf; and (4) record weight gain and footprint size throughout the first year.

Non-invasive detection of candidate pregnancy protein biomarkers in the feces of captive polar bears (Ursus maritimus).

Currently, there is no method of accurately and non-invasively diagnosing pregnancy in polar bears. Specific proteins may exhibit altered profiles in the feces of pregnant bears, but predicting appropriate candidate proteins to investigate is speculative at best. The objective of this study was to identify potential pregnancy biomarker proteins based on their increased abundance in the feces of pregnant polar bears compared to pseudopregnant females (controls) using two-dimensional in-gel electrophoresis (2D-DIGE) and mass spectrometry (MS).

Attempted in vitro maturation and fertilization of postmortem Sumatran rhinoceros (Dicerorhinus sumatrensis) oocytes.

A study was conducted opportunistically to evaluate the potential of rescuing immature oocytes from the ovaries of the Sumatran rhinoceros postmortem. Recovered oocytes (n = 30) were placed in maturation culture for 36 hr and inseminated with frozen-thawed homologous spermatozoa. After culture, evaluation of nuclear maturation status revealed that a large number of oocytes were degenerated (n = 21), but nine oocytes were assessed at the germinal vesicle (n = 3), metaphase I (n = 3), and metaphase II (n = 3) stages.

Gamete rescue in the African black rhinoceros (Diceros bicornis).

Mortality rates are high among captive African black rhinoceroses (Diceros bicornis), due to increased susceptibility to disease. The ability to rescue genetic material from individuals that die unexpectedly represents a practical approach to assist ex situ conservation efforts. The objectives of the present study were to attempt postmortem oocyte recovery from ovaries of African black rhinoceroses (N = 6) and to test the efficacy of equine protocols for rhinoceros oocyte IVM and IVF using cryopreserved rhinoceros sperm.

Pulmonary adenocarcinoma and endogenous lipid pneumonia in a common genet (Genetta genetta).

An adult male common genet (Genetta genetta) was examined for an onset of severe dyspnea. Physical examination findings included tachypnea and increased inspiratory sounds. Survey radiographs showed consolidation of both lung lobes, and a bronchoalveolar lavage sample contained cholesterol crystals.

Mixed infection with reovirus and Chlamydophila in a flock of budgerigars (Melopsittacus undulatus).

Eleven budgerigars (Melopsittacus undulatus) from a zoological collection presented at necropsy with emaciation and splenomegaly or hepatomegaly or both. Polymerase chain reaction assays performed on liver and spleen samples were positive for Chlamydophila psittaci in 2 of 3 birds tested, and histologic findings in 2 additional birds were compatible with chlamydiosis. The aviary was subsequently closed to the public, and a 45-day treatment regimen with doxycycline in the seeds was initiated.

Alkaline phosphatase as an indicator of true ejaculation in the rhinoceros.

The objective was to determine if seminal alkaline phosphatase (ALP) can serve as an indicator of true ejaculation in the rhinoceros. Concentrations of ALP activity were determined in seminal fractions collected from African black rhinos (Diceros bicornis), an African white rhino (Ceratotherium simum), and an Indian rhino (Rhinoceros unicornis) during electroejaculation. In addition, seminal fractions collected during penile massage of a Sumatran rhino (Dicerorhinus sumatrensis) were assessed.

Complete sequence and gene organization of the mitochondrial genome for Hubbard's sportive lemur (Lepilemur hubbardorum).

The complete mitochondrial DNA (mtDNA) genome of Hubbard's or Zombitse sportive lemur (Lepilemur hubbardorum) was generated by polymerase chain reaction (PCR) amplification, primer-walking sequencing and fragment cloning. Comparative analyses of Hubbard's sportive lemur were conducted with available complete mitochondrial genome sequences from eight other lemur species. The mitochondrial genome of Hubbard's sportive lemur is 16,854 base pairs (bp) and contains 13 protein-coding genes, 22 transfer RNA (tRNA) genes, two ribosomal RNA (rRNA) genes and one control region.

Gastrointestinal obstruction in penguin chicks.

A 7-day-old gentoo penguin (Pygoscelis papua) was found dead and postmortem examination revealed impaction of the ventriculus with feathers. A review of mortality in gentoo penguin chicks from 1997 to 2007 at that institution revealed another case of feather impaction of the ventriculus in a 4-week-old chick, a sibling of the previous chick. A third case of gastrointestinal impaction occurred in a 24-day-old king penguin (Aptenodytes patagonicus) with omphallitis and enteritis.

Semen cryopreservation in the Indian rhinoceros (Rhinoceros unicornis).

The objective was to identify an extender and cryoprotectant combination for Indian rhinoceros (Rhinoceros unicornis) sperm that yielded high post-thaw sperm quality. Male Indian rhinoceroses (n=6; 7.5-34 yr old) were anesthetized and subjected to a regimented electroejaculation procedure (75-100 mAmps; 4-10 volts; 7-150 stimuli; total of 10 electroejaculation procedures).

Effects of age, weight, hormones, and hibernation on breeding success in boreal toads (Bufo boreas boreas).

The goals of this study were to test the effects of exogenous hormones and hibernation on breeding behavior and gamete release by boreal toads (Bufo boreas boreas). Each year, a subset of 77 toads was hibernated and then paired with hibernated or nonhibernated mates and treated with luteinizing hormone releasing hormone analogue (LHRHa), human chorionic gonadotropin (hCG), or left untreated. Amplexus and egg and sperm production were recorded.

Detection of cytonuclear genomic dissociation in the North American captive African elephant collection.

A total of 114 captive elephants (6 Asian; 108 African) from 43 private institutions or North American zoos accredited by the Association of Zoos and Aquariums were sampled and evaluated to investigate genetic status. Because previous analyses of the captive collection indicated potential cytonuclear dissociation between mitochondrial DNA (mtDNA) sequence and microsatellite nuclear DNA genotype data, we investigated this phenomenon within the captive collection with 2 X-linked genes (BGN and PHKA2) and 1 Y-linked gene (AMELY). These data reveal that individuals with forest-derived elephant mtDNA lineages carried only savannah elephant nuclear gene haplotypes.

Characterization of basal seminal traits and reproductive endocrine profiles in North American river otters and Asian small-clawed otters.

In this study, fecal samples were collected from 24 North American river (NARO) and 17 Asian small-clawed otters (ASCO) for 6-36 months and semen collected seasonally from NARO males (n=4/season) via electroejaculation. Our main objectives were to: (1) characterize endocrine parameters by longitudinal monitoring of fecal hormone metabolites and (2) investigate semen collection and basal seminal traits in NARO. NARO demonstrated a distinct seasonality in the spring, with females having a monoestrual estrogen elevation lasting 15.

Evaluating the utility of an accelerometer and urinary hormone analysis as indicators of estrus in a Zoo-housed koala (Phascolarctos cinereus).

Induced ovulators, such as the koala, do not always have overt signs of estrus, which makes pairing these animals for breeding purposes difficult to achieve in a zoo setting. This study examined the possibility of using alternative methods to behavioral sampling and weight fluctuations for monitoring estrus in a female koala of reproductive age. We attempt to gain an improved understanding of koala estrus and enhance our ability to detect it by combining a noninvasive technique for hormone analysis with a newer method for recording activity level.

Technical note: effect of density gradient centrifugation with trypsin on the in vivo fertilising capability of bovine spermatozoa.

The present study investigated the effect of a novel density gradient centrifugation (DGC) treatment using recombinant trypsin on the in vivo fertilising capability of bovine spermatozoa compared with a standard method. In Trial 1, semen collected from Boran and Ankole (Bos indicus) bulls was treated either with a silane-coated silica particle colloid formulated for humans with a recombinant trypsin or processed using a standard method (dilution in an egg yolk-based diluent). Semen processed by the two methods was used to artificially inseminate (AI) superovulated cattle.

Urinalysis in three species of captive rhinoceros (Rhinoceros unicornis, Dicerorhinus sumatrensis, and Diceros bicornis).

This study reports urinalysis values for three species of captive rhinoceros (Rhinoceros unicornis, Dicerorhinus sumatrensis, and Diceros bicornis) and evaluates individual and species differences. Repeated urinalysis was conducted on 11 individuals to establish normal reference ranges. Although no individual or species differences existed in urinary values for pH, all species differed in specific gravity.