Laurie Marker

Phylogenetics, genome diversity and origin of modern leopard, Panthera pardus

Leopards, Panthera pardus, are widely distributed across southern Asia and sub‐Saharan Africa. The extent and phylogeographic patterns of molecular genetic diversity were addressed in a survey of 77 leopards from known geographical locales representing 13 of the 27 classical trinomial subspecies. Phylogenetic analysis of mitochondrial DNA sequences (727 bp of NADH5 and control region) and 25 polymorphic microsatellite loci revealed abundant diversity that could be partitioned into a minimum of nine discrete populations, tentatively named here as revised subspecies: P. parduspardus, P. p. nimr, P. p. saxicolor, P. p. fusca, P. p. kotiya, P. p. delacouri, P. p. japonensis, P. p. orientalis and P. p. melas. However, because of limited sampling of African populations, this may be an underestimate of modern phylogeographic population structure. Combined phylogeographic and population diversity estimates support an origin for modern leopard lineages 470 000–825 000 years ago in Africa followed by their migration into and across Asia more recently (170 000–300 000 years ago). Recent demographic reductions likely have led to genetic impoverishment in P. p. orientalis and in the island subspecies P. p. kotiya.

EVIDENCE FOR CHRONIC STRESS IN CAPTIVE BUT NOT FREE- RANGING CHEETAHS (ACINONYX JUBATUS) BASED ON ADRENAL MORPHOLOGY AND FUNCTION

The cheetah (Acinonyx jubatus)is highly endangered because of loss of habitat in the wild and failure to thrive in captivity. Cheetahs in zoos reproduce poorly and have high prevalences of unusual diseases that cause morbidity and mortality. These diseases are rarely observed in free-ranging cheetahs but have been documented in cheetahs that have been captured and held in captive settings either temporarily or permanently. Because captivity may be stressful for this species and stress is suspected as contributing to poor health and reproduction, this study aimed to measure chronic stress by comparing baseline concentrations of fecal corticoid metabolites and adrenal gland morphology between captive and free-ranging cheetahs. Additionally, concentrations of estradiol and testosterone metabolites were quantified to determine whether concentrations of gonadal steroids correlated with corticoid concentration and to assure that corticosteroids in the free-ranging samples were not altered by environmental conditions. Concentrations of fecal corticoids, estradiol, and testosterone were quantified by radioimmunoassay in 20 free-ranging and 20 captive cheetahs from samples collected between 1994 and 1999. Concentrations of baseline fecal corticoids were significantly higher (p=0.005) in captive cheetahs (196.08±36.20 ng/g dry feces) than free-ranging cheetahs (71.40±14.35 ng/g dry feces). Testosterone concentrations were lower in captive male cheetahs (9.09±2.84 ng/g dry feces) than in free-ranging cheetahs (34.52±12.11 ng/g dry feces), which suggests suppression by elevated corticoids in the captive males. Evidence for similar suppression of estradiol concentrations in females was not present. Adrenal corticomedullary ratios were determined on midsagittal sections of adrenal glands from 13 free-ranging and 13 captive cheetahs obtained between 1991 and 2002. The degree of vacuolation of cortical cells in the zona fasciculata was graded for each animal. Corticomedullary ratios were larger (p=0.05) in captive cheetahs; however, there was no difference (p=0.31) in the degree of corticocyte vacuolation between the two populations. These data provide both morphologic and functional evidence suggestive of chronic stress in captive cheetahs. Further research into the role of hypercortisolemia in the pathogenesis of the reproductive abnormalities and unusual diseases of captive cheetahs is needed.

Extrinsic factors significantly affect patterns of disease in free-ranging and captive cheetah (Acinonyx jubatus) populations

The cheetah (Acinonyx jubatus) has been considered a paradigm for disease vulnerability due to loss of genetic diversity. This species monomorphism has been suspected to be the basis for their general poor health and dwindling populations in captivity. North American and South African captive populations have high prevalences of hepatic veno-occlusive disease, glomerulosclerosis, gastritis, and systemic amyloidosis, diseases that are rare in other species. Unusually severe inflammatory reactions to common infectious agents have also been documented in captive cheetahs. The current study compared disease prevalences in free-ranging Namibian cheetahs with those in two captive populations of similar ages. The occurrence of diseases in the free-ranging population was determined from 49 necropsies and 27 gastric biopsies obtained between 1986 and 2003 and compared with prevalences in 147 North American and 80 South African captive cheetahs. Except for two cheetahs, the free-ranging population was in robust health with only mild lesions present, in contrast with significantly higher prevalences in the captive populations. Despite widespread heavy Helicobacter colonization in wild cheetahs, only 3% of the free-ranging population had moderate to severe gastritis, in contrast with 64% of captive cheetahs. No severe inflammatory reactions to viral infections were detected in the free-ranging animals. Because free-ranging Namibian cheetahs are as genetically impoverished as captive cheetahs, these findings caution against attributing loss of fitness solely to genetic factors and attest to the fundamental importance of extrinsic factors in wildlife health.

Aspects of the management of cheetahs, Acinonyx jubatus jubatus, trapped on Namibian farmlands

Abstract The Namibian cheetah population has recently undergone serious decline due to human-mediated removals, and investigating the rates and causes of such removals is an important aspect of the future management of cheetah populations outside protected areas. We examined cheetahs that were reported live-trapped or killed on Namibian farmlands between 1991 and 1999. A perceived threat to livestock or game led to the vast majority of live captures and to almost half of the cheetah deaths investigated. Despite this, livestock predation from cheetahs appeared to be minimal, and was usually perpetrated by cheetahs with injuries. Most of the cheetahs were trapped in groups, and cheetahs’ relative sociality leads to the easy removal of entire social units. Long-term monitoring must include detailed consideration of these indiscriminate removals, as they involve many cheetahs, fluctuate between years, often go unreported, and are likely to have a serious impact on cheetah populations outside protected areas.

Captive breeding of the cheetah (Acinonyx jubatus) in North American zoos (1871–1986)

The African cheetah has been bred in North American zoological facilities since 1956. The captive population has since grown to around 200 animals because of a combined increase in importation plus captive births. From 1982 to 1986, the captive birth rate declined by 50%, primarily because of a low frequency of breeding individuals in the population. The 1986 population had an effective breeding size of 28.1 in a total population of over 193 cheetahs. The incidence of infant mortality has been high (36.7%) relative to other zoo-bred species, perhaps as a consequence of the previously observed genetic impoverishment of the species. The combination of low fecundity, high infant mortality, and population dynamics indicates that the North American captive cheetah population is neither a self-sustaining nor a theoretically “viable population” as defined by Soule et al. [ZOO BIOLOGY 5:101–114, 1986]. Possible recommendations for improving captive cheetah propagation are discussed.

Specifying and sustaining pigmentation patterns in domestic and wild cats

What Kitty Shares with Kings Although long-studied, the underlying basis of mammalian coat patterns remains unclear. By studying a large number of cat species and varieties, Kaelin et al. (p. 1536) identified two genes, Taqpep and Edn3, as critical factors in the development of feline pigment patterns. Mutations in Taqpep are responsible for the blotched tabby pattern in domestic cats and the unusual coat of wild king cheetahs. Gene expression patterns in cat and cheetah skin suggest that Edn3 is a likely regulator of felid hair color. The findings support a common model for coat and pigment pattern formation in domestic and wild cats. The genes specifying tabby cat coat patterns also affect big cats, including king cheetahs. Color markings among felid species display both a remarkable diversity and a common underlying periodicity. A similar range of patterns in domestic cats suggests a conserved mechanism whose appearance can be altered by selection. We identified the gene responsible for tabby pattern variation in domestic cats as Transmembrane aminopeptidase Q (Taqpep), which encodes a membrane-bound metalloprotease. Analyzing 31 other felid species, we identified Taqpep as the cause of the rare king cheetah phenotype, in which spots coalesce into blotches and stripes. Histologic, genomic expression, and transgenic mouse studies indicate that paracrine expression of Endothelin3 (Edn3) coordinates localized color differences. We propose a two-stage model in which Taqpep helps to establish a periodic pre-pattern during skin development that is later implemented by differential expression of Edn3.

Ejaculate traits in the Namibian cheetah (Acinonyx jubatus): influence of age, season and captivity

The objective was to examine the influence of animal age, season and captivity status on seminal quality in wild-born cheetahs (Acinonyx jubatus) in Namibia, Africa. Animals were divided into three age categories: juvenile (14-24 months; n = 16 males, 23 ejaculates); adult (25-120 months; n = 76 males, 172 ejaculates); and aged (>120 months; n = 5 males, 5 ejaculates). Seasons were categorised into hot-wet (January-April), cold-dry (May-August) and hot-dry (September-December). A comparison between freshly wild-caught (n = 29 males, 41 ejaculates) and captive-held cheetahs (n = 68 males, 159 ejaculates) was also conducted. Raw ejaculates contained 69.0 +/- 1.1% motile spermatozoa (mean +/- s.e.m.) with 73.6 +/- 1.5% of these cells containing an intact acrosome. Overall, 18.4 +/- 0.9% of spermatozoa were morphologically normal, with midpiece anomalies being the most prevalent (approximately 39%) defect. Juvenile cheetahs produced ejaculates with poorer sperm motility, forward progressive status, lower seminal volume and fewer total motile spermatozoa than adult and aged animals. Spermatogenesis continued unabated throughout the year and was minimally influenced by season. Proportions of sperm malformations were also not affected by season. Ejaculates from captive cheetahs had increased volume and intact acrosomes, but lower sperm density than wild-caught counterparts. In summary, Namibian cheetahs produce an extraordinarily high proportion of pleiomorphic spermatozoa regardless of age, season or living (captive versus free-ranging) status. Young males less than 2 years of age produce poorer ejaculate quality than adult and aged males. Because (1) all study animals were wild born and (2) there was little difference between freshly caught males and those maintained in captivity for protracted periods, our results affirm that teratospermia in the cheetah is mostly genetically derived. It also appears that an ex situ environment for the Namibian cheetah can ensure sperm quality comparable with that for free-living males.

SEROSURVEY OF VIRAL INFECTIONS IN FREE-RANGING NAMIBIAN CHEETAHS (ACINONYX JUBATUS)

Cheetahs (Acinonyx jubatus) in captivity have unusually high morbidity and mortality from infectious diseases, a trait that could be an outcome of population homogeneity or the immunomodulating effects of chronic stress. Free-ranging Namibian cheetahs share ancestry with captive cheetahs, but their susceptibility to infectious diseases has not been investigated. The largest remaining population of free-ranging cheetahs resides on Namibian farmlands, where they share habitat with domestic dogs and cats known to carry viruses that affect cheetah health. To assess the extent to which free-ranging cheetahs are exposed to feline and canine viruses, sera from 81 free-ranging cheetahs sampled between 1992 and 1998 were evaluated for antibodies against canine distemper virus (CDV), feline coronavirus (feline infectious peritonitis virus; FCoV/FIPV), feline herpesvirus 1 (FHV1), feline panleukopenia virus (FPV), feline immunodeficiency virus (FIV), and feline calicivirus (FCV) and for feline leukemia virus (FeLV) antigens. Antibodies against CDV, FCoV/FIPV, FHV1, FPV, and FCV were detected in 24, 29, 12, 48, and 65% of the free-ranging population, respectively, although no evidence of viral disease was present in any animal at the time of sample collection. Neither FIV antibodies nor FeLV antigens were present in any free-ranging cheetah tested. Temporal variation in FCoV/FIPV seroprevalence during the study period suggested that this virus is not endemic in the free-ranging population. Antibodies against CDV were detected in cheetahs of all ages sampled between 1995 and 1998, suggesting the occurrence of an epidemic in Namibia during the time when CDV swept through other parts of sub-Saharan Africa. This evidence in free-ranging Namibian cheetahs of exposure to viruses that cause severe disease in captive cheetahs should direct future guidelines for translocations, including quarantine of seropositive cheetahs and preventing contact between cheetahs and domestic pets.

Demography of the Namibian cheetah, Acinonyx jubatus jubatus

Abstract Namibian cheetahs have suffered, and continue to suffer, high levels of removal due to conflict with local farmers, and it is important to understand the demography of this population in order to determine its likely persistence. Examination of cheetahs reported live-trapped or killed by local farmers, combined with subsequent information from radio-telemetry, allowed demographic parameters such as sex ratios, age and social structure, litter size, interbirth intervals and survivorship to be estimated for cheetahs on Namibian farmlands. Cub mortality was relatively low, but adult mortality was high, particularly for males, and peaked at 5–6 years of age. Neither marking nor relocating cheetahs seemed to affect survivorship, and there was no difference in survivorship between the sexes. Time spent in captivity did not appear to affect survival after release. These findings will be useful in formulating recommendations regarding the conservation and sustainable utilization of cheetah populations outside protected areas.

MORPHOLOGY, PHYSICAL CONDITION, AND GROWTH OF THE CHEETAH (ACINONYX JUBATUS JUBATUS )

Abstract Information regarding morphology of wild cheetahs is scant, and even where data exist they rarely were collected using a standardized methodology. We used a consistent technique to examine 241 wild Namibian cheetahs (Acinonyx jubatus jubatus) to study morphology, sexual dimorphism, growth rates, and physical condition and to investigate how these data compared with those in previous studies. Significant sexual dimorphism was evident for all measurements. The majority of cheetahs were in excellent condition at the time of examination, although old cheetahs and those that had been held captive for more than a month were in significantly poorer condition. Both male and female cheetahs reached adult body mass at 49–96 months of age. These data differed significantly from those collected during other studies, although such differences may be due to variations in collection methodology. It is therefore vital to standardize morphometric data collection techniques so that the true extent of differences between populations can be assessed more accurately. A suggested standardized collection methodology is presented.