Linda M. Penfold

Genetic Restoration of the Florida Panther

Return of the Florida Panther The Florida panther is an endangered lineage that has been the target of extensive human management to maintain genetic diversity within a small population. Now, Johnson et al. (p. 1641; see the Perspective by Packer) provide comprehensive genetic and demographic analyses of a project to rescue the Florida panther from inbreeding, low genetic diversity, inbreeding depression, and demographic stochasticity by outcrossing it to Texas pumas. The genetics and reproductive fitness traits of different panther populations and their crosses reveal the benefits of outcrossing on survival, genetic diversity, male reproductive characteristics, and population demography. Recovery of the Florida panther population is attributed to the benefits of admixture with panthers from Texas. The rediscovery of remnant Florida panthers (Puma concolor coryi) in southern Florida swamplands prompted a program to protect and stabilize the population. In 1995, conservation managers translocated eight female pumas (P. c. stanleyana) from Texas to increase depleted genetic diversity, improve population numbers, and reverse indications of inbreeding depression. We have assessed the demographic, population-genetic, and biomedical consequences of this restoration experiment and show that panther numbers increased threefold, genetic heterozygosity doubled, survival and fitness measures improved, and inbreeding correlates declined significantly. Although these results are encouraging, continued habitat loss, persistent inbreeding, infectious agents, and possible habitat saturation pose new dilemmas. This intensive management program illustrates the challenges of maintaining populations of large predators worldwide.

Artificial insemination in deer and non-domestic bovids

Artificial insemination technology has revolutionized the domestic cattle breeding industry and allowed for the dissemination of valuable genetics worldwide. This technology has been adapted for use in many other taxa for the conservation of threatened and endangered species, but its use for the genetic management of small populations of deer, antelope and other non-domestic bovids has met numerous challenges and limited success. In practice, adaptation of domestic bovine AI protocols to other artiodactylids for genetic management has been limited by: (1) a lack of understanding of species-specific reproductive characteristics; (2) the inability to minimize handling stress; (3) pregnancy losses; and (4) regulatory challenges in semen importation. To date, AI protocols have been developed for seven species of cervid and 14 species of non-domestic bovids; recent developments in this technology has allowed greater use of AI for dissemination of genetics in farmed deer species. However, despite decades of research in the use of assisted reproduction for the conservation of antelope and other non-domestic bovids, even this simplest technique has not been used repeatedly for genetic management.

Seasonal patterns of LH, testosterone and semen quality in the Northern pintail duck (Anas acuta).

This study characterized seasonal changes in circulating LH and testosterone and in semen production and quality in the Northern pintail duck. Plasma LH and testosterone were measured in blood samples collected weekly throughout the year from eight males exposed to natural fluctuations in day length and temperature. Semen quality was evaluated weekly in these same males from April-June, the months when spermatozoa were produced. Semen quality (based on sperm concentration and normal morphology) peaked 0-2 weeks after sperm production onset and decreased sharply before sperm production cessation in late June. Nadir LH concentrations were measured in July and August with peak LH observed in May and November. There were clear seasonal patterns in circulating testosterone with July-September values being less (P<0.05) than October-December which, in turn, were less (P<0.05) than January-March. Maximal circulating testosterone (P<0.05) occurred during April-June, coincident with semen production. Weekly circulating LH during the breeding season was directly related to testosterone concentrations (P<0.01), but was not correlated to any specific semen or sperm trait (P>0.05). Testosterone concentrations throughout the breeding season were correlated (P<0.05) to total numbers of spermatozoa produced (volume x cell concentration) and percent normal sperm morphology. In summary, the Northern pintail experiences seasonal hormone fluctuations, with maximum circulating testosterone coinciding with peak ejaculate quality reflected by the production of high numbers of morphologically normal spermatozoa.

Use it or lose it: Characterization, implications, and mitigation of female infertility in captive wildlife

Zoos and other ex situ wildlife institutions can play an important role in species conservation by maintaining populations for education and research, as sources for potential re-introduction or reinforcement, and as ambassadors for financial support of in situ conservation. However, many regional zoo associations are realizing that current captive populations are unsustainable, with many programs failing to meet demographic and genetic goals to ensure long-term viability. Constraints on population size due to limited space often mandate delayed and/or less frequent breeding, but for females of many species this can have profound effects on fertility. A retrospective analysis combined with published literature and reliable anecdotal reports reveals that, when females are housed in a non-breeding situation for extended periods of time, reproductive changes that negatively impact fertility have occurred in multiple species, including canids, elephants, white rhinoceros, Sebas bats, wildebeest, stingrays, and some felid species. Competing space needs and changing interest in taxa for exhibits over time compound the problem. Counter strategies to breed early and often have their own demographic and genetic consequences as well as logistical and political implications. Strategies to mitigate the sustainability crisis in these taxa might include a mixed strategy in which young, genetically valuable females are bred earlier and at more regular intervals to ensure reproductive success, in combination with the judicious use of available tools to manage the number of offspring produced, including contraception and culling. An understanding of the issues at stake is the first step towards developing management strategies for sustainable populations.

Using PGFM (13,14-dihydro-15-keto-prostaglandin F2α) as a non-invasive pregnancy marker for felids

Understanding the complex endocrine interactions that control reproduction in felids is essential for captive breeding management. The most important demand is a quick and reliable pregnancy diagnosis. However, the occurrence of pseudopregnancies in felids complicates matters. We investigated whether the fecal prostaglandin metabolite (PGFM) recently suggested for pregnancy diagnosis in the lynx is suitable for all felid species. We found that increased levels of PGFM during the last trimester indicate pregnancy in seven of the eight main lineages of the carnivore family Felidae. PGFM levels in a sand cat (domestic cat lineage) were basal at mating and remained so until Day 40 post-mating. Day 41 marked the beginning of a distinct increase culminating in peak levels of 6.5 μg/g before parturition and decreasing again to baseline thereafter. Similar pregnancy profiles were obtained from the domestic cat, the leopard cat, the lynx, the ocelot and the caracal lineage, whereas in pseudopregnant individuals (sand cat, Iberian and Eurasian lynx) fecal PGFM remained at basal levels. In pregnant cheetahs (puma lineage) PGFM increased above basal following day ∼48 peaking before pregnancy but remained at baseline in pseudopregnant females. Discrepancies existed in the Panthera lineage. While Chinese leopard, Sumatran tiger, and the black panther showed marked increases of PGFM during the last weeks of pregnancy, only moderate increases in PGFM levels were found in the Indochinese tiger and the Persian leopard. Altogether, PGFM as tool for pregnancy diagnosis has been proven to be useful in breeding management of felids.

International Society for Wildlife Endocrinology: the future of endocrine measures for reproductive science, animal welfare and conservation biology

Hormone analysis is a precise and widely accepted tool for monitoring reproductive function and responses to stressors. Although hormones are present and can be measured in various biological matrices, non-invasive methods have gained popularity over the past 30 years as a more practical approach for assessing ovarian, testicular and, more recently, adrenocortical activity in intractable wildlife species. Non-invasive hormone monitoring also has been key to understanding biological mechanisms related to observed behaviours of captive and free-ranging animals. Despite the increasing popularity of this research field, wildlife endocrinologists have not had a specific forum for sharing and discussing their latest findings, technical developments and common challenges. To provide such a communication platform, the International Society for Wildlife Endocrinology (ISWE) was established in 2010, followed by an international meeting held on 3–4 November 2011 at the Toronto Zoo, Canada. Over several sessions, keynote speakers and participants discussed recent developments of new and innovative methods for hormone monitoring, as well as the latest advances in basic endocrinology as applied to adrenal function, reproductive physiology, animal health, ecology and evolution. Here, we introduce ISWE to the scientific community and discuss how this new society will serve as a resource for wildlife endocrinologists worldwide.

Evidence for Compromised Metabolic Function and Limited Glucose Uptake in Spermatozoa from the Teratospermic Domestic Cat (Felis catus) and Cheetah (Acinonyx jubatus)

Cheetahs and certain other felids consistently ejaculate high proportions (≥60%) of malformed spermatozoa, a condition known as teratospermia, which is prevalent in humans. Even seemingly normal spermatozoa from domestic cat teratospermic ejaculates have reduced fertilizing capacity. To understand the role of sperm metabolism in this phenomenon, we conducted a comparative study in the normospermic domestic cat versus the teratospermic cat and cheetah with the general hypothesis that sperm metabolic function is impaired in males producing predominantly pleiomorphic spermatozoa. Washed ejaculates were incubated in chemically defined medium containing glucose and pyruvate. Uptake of glucose and pyruvate and production of lactate were assessed using enzyme-linked fluorescence assays. Spermatozoa from domestic cats and cheetahs exhibited similar metabolic profiles, with minimal glucose metabolism and approximately equimolar rates of pyruvate uptake and lactate production. Compared to normospermic counterparts, pyruvate and lactate metabolism were reduced in teratospermic cat and cheetah ejaculates, even when controlling for sperm motility. Rates of pyruvate and lactate (but not glucose) metabolism were correlated positively with sperm motility, acrosomal integrity, and normal morphology. Collectively, our findings reveal that pyruvate uptake and lactate production are reliable, quantitative indicators of sperm quality in these two felid species and that metabolic function is impaired in teratospermic ejaculates. Furthermore, patterns of substrate utilization are conserved between these species, including the unexpected lack of exogenous glucose metabolism. Because glycolysis is required to support sperm motility and capacitation in certain other mammals (including dogs), the activity of this pathway in felid spermatozoa is a target for future investigation.

Review of contraception in ungulate species

Most ungulate species are herd animals. In captivity, and increasingly so in the wild, space constraints limit natural behaviors associated with group dynamics, possibly resulting in inbreeding and/or overpopulation. This situation has necessitated research regarding contraception of various species of hoofstock. Differing management situations mandate different contraception protocols to achieve optimal results. Fertility control in hoofstock has been achieved through a number of different contraceptive methods predominantly surgical sterilization, mechanical contraception, synthetic steroid hormones, and immunocontraception. In this study successes and limitations of these techniques are reviewed. Zoo Biol 26:311-326, 2007. (c) 2007 Wiley-Liss, Inc.

Glycolytic Enzyme Activity Is Essential for Domestic Cat (Felis catus) and Cheetah (Acinonyx jubatus) Sperm Motility and Viability in a Sugar-Free Medium

We have previously reported a lack of glucose uptake in domestic cat and cheetah spermatozoa, despite observing that these cells produce lactate at rates that correlate positively with sperm function. To elucidate the role of glycolysis in felid sperm energy production, we conducted a comparative study in the domestic cat and cheetah, with the hypothesis that sperm motility and viability are maintained in both species in the absence of glycolytic metabolism and are fueled by endogenous substrates. Washed ejaculates were incubated in chemically defined medium in the presence/absence of glucose and pyruvate. A second set of ejaculates was exposed to a chemical inhibitor of either lactate dehydrogenase (sodium oxamate) or glyceraldehyde-3-phosphate dehydrogenase (alpha-chlorohydrin). Sperm function (motility and acrosomal integrity) and lactate production were assessed, and a subset of spermatozoa was assayed for intracellular glycogen. In both the cat and cheetah, sperm function was maintained without exogenous substrates and following lactate dehydrogenase inhibition. Lactate production occurred in the absence of exogenous hexoses, but only if pyruvate was present. Intracellular glycogen was not detected in spermatozoa from either species. Unexpectedly, glycolytic inhibition by alpha-chlorohydrin resulted in an immediate decline in sperm motility, particularly in the domestic cat. Collectively, our findings reveal an essential role of the glycolytic pathway in felid spermatozoa that is unrelated to hexose metabolism or lactate formation. Instead, glycolytic enzyme activity could be required for the metabolism of endogenous lipid-derived glycerol, with fatty acid oxidation providing the primary energy source in felid spermatozoa.

Effects of sociosexual environment on serum testosterone in captive male African rhinoceros.

The relationships between testosterone concentrations in male African rhinoceros and the presence of conspecific males and females were investigated. Serum testosterone concentrations were measured using enzyme-linked immunoassay (EIA) in 37 male black rhinoceros (Diceros bicornis) and 21 male white rhinoceros (Ceratotherium simum) housed at 37 institutions in the USA. Testosterone concentrations in both black (n=37) and white (n=21) rhinoceros males rose with increasing numbers of females present (P<0.05). Average testosterone concentrations also rose with an increased number of conspecific males (n=34) in black rhinoceros (P<0.05). However, no specific pattern was found among male white rhinoceros housed with other males. We inferred that introduction of females to a male may play an important role in stimulating libido and spermatogenesis. The similar response of black rhinoceros and white rhinoceros to increased numbers of females suggested that, at least historically, herd structure for blacks may have been more similar to whites than previously realized, and should be investigated further.