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Feline immunodeficiency virus (FIV) infects numerous wild and domestic feline species and is closely related to human immunodeficiency virus (HIV) and simian immunodeficiency virus (SIV). Species-specific strains of FIV have been described for domestic cat (Felis catus), puma (Puma concolor), lion (Panthera leo), leopard (Panthera pardus), and Pallas' cat (Otocolobus manul). Here, we employ a three-antigen Western blot screening (domestic cat, puma, and lion FIV antigens) and PCR analysis to survey worldwide prevalence, distribution, and genomic differentiation of FIV based on 3,055 specimens from 35 Felidae and 3 Hyaenidae species. Although FIV infects a wide variety of host species, it is confirmed to be endemic in free-ranging populations of nine Felidae and one Hyaenidae species. These include the large African carnivores (lion, leopard, cheetah, and spotted hyena), where FIV is widely distributed in multiple populations; most of the South American felids (puma, jaguar, ocelot, margay, Geoffroy's cat, and tigrina), which maintain a lower FIV-positive level throughout their range; and two Asian species, the Pallas' cat, which has a species-specific strain of FIV, and the leopard cat, which has a domestic cat FIV strain in one population. Phylogenetic analysis of FIV proviral sequence demonstrates that most species for which FIV is endemic harbor monophyletic, genetically distinct species-specific FIV strains, suggesting that FIV transfer between cat species has occurred in the past but is quite infrequent today.

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. pardus pardus, 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.

Sperm-oocyte interaction in vitro was studied in the cheetah, a species known to produce poor quality ejaculates and to experience low rates of fertility. Twelve female cheetahs were injected (i.m.) with eCG followed by hCG 84 h later. Twenty-four to 26 h post hCG, each was subjected to laparoscopic oocyte aspiration. A sperm motility index (SMI) was calculated for each of 9 cheetah sperm donors that produced ejaculates averaging 41.3 +/- 22.9 x 10(6) motile sperm and 28.4 +/- 4.9% structurally normal sperm. Each ejaculate was used to inseminate cheetah oocytes from 1 or 2 females and salt-stored, domestic cat oocytes. The presence of ovarian follicles (greater than or equal to 1.5 mm in diameter) showed that all females responded to exogenous gonadotropins (range, 11-35 follicles/female). A total of 277 cheetah oocytes was collected from 292 follicles (94.9% recovery; 23.1 +/- 2.2 oocytes/female). Of these, 250 (90.3%) qualified as mature and 27 (9.7%) as degenerate. Of the 214 mature oocytes inseminated, 56 (26.2%) were fertilized, and 37 (17.3%) cleaved to the 2-cell stage in vitro; but the incidence of in vitro fertilization (IVF) varied from 0 to 73.3% (p less than 0.001) among individual males. When oocytes from individual cheetahs (n = 5) were separated into two groups and inseminated with sperm from a male with an SMI greater than 0 after 6 h coincubation versus an SMI = 0 at this time, the mean fertilization rates were 28/44 (63.6%) and 0/37 (0%), respectively (p less than 0.05). Of the 117 domestic cat oocytes coincubated with cheetah sperm, 96.6% contained 1 or more cheetah sperm in the outer half of the zona pellucida (ZP). Although the mean number of cheetah sperm penetrating the outer ZP of the cat oocyte was similar (p greater than 0.05) among all males, there was a positive correlation between the number of sperm reaching the inner half of the ZP and fertilization rate in vitro (r = 0.82; p less than 0.05). Compared to IVF efficiency in the domestic cat and tiger as reported in earlier studies, IVF efficiency in the cheetah is low. Because oocytes from 11 of 12 cheetahs were fertilized in vitro, there is no evidence that the female gamete is incompetent. Although sperm pleiomorphisms may contribute to poor reproductive performance, examination of the data on the basis of individual sperm donors reveals that effective gamete interaction in the cheetah is dictated, in part, by sperm motility.

Pituitary-gonadal function was examined in male lions free-ranging in the Serengeti Plains or geographically isolated in the Ngorongoro Crater of Tanzania. Lions were classified by age as adult (6.1-9.8 years), young adult (3.3-4.5 years) or prepubertal (1.4-1.6 years, Serengeti Plains only). Each animal was anaesthetized and then bled at 5-min intervals for 100 min before and 140 min after i.v. administration of saline or GnRH (1 micrograms/kg body weight). Basal serum LH and FSH concentrations were similar (P greater than 0.05) among age classes and between locations. In Serengeti Plains lions, net LH peak concentrations after GnRH were approximately 35% greater (P less than 0.05) in prepubertal than in either adult or young adult animals. GnRH-stimulated LH release was similar (P greater than 0.05) between adult and young adult lions, and these responses were similar (P greater than 0.05) to those measured in Ngorongoro Crater lions. Basal and GnRH-stimulated testosterone secretion was higher (P less than 0.05) in adult than in young adult lions and lowest (P less than 0.05) in prepubertal lions. Age-class differences in testosterone production were related directly to the concentrations of LH receptors in the testis (P less than 0.05). Basal and GnRH-stimulated testosterone secretion and gonadotrophin receptor concentrations within age classes were similar (P greater than 0.05) between lions of the Serengeti Plains and Ngorongoro Crater. Lower motility and higher percentages of structurally abnormal spermatozoa were observed in electroejaculates of young adult compared to adult Serengeti Plains males (P less than 0.05) and were associated with decreased steroidogenic activity. In contrast, there were no age-related differences in ejaculate characteristics of Ngorongoro Crater lions. Seminal quality in the Crater population was poor in adult and young adult animals and was unrelated to alterations in pituitary or testicular function. In summary, only seminal quality in adult male lions was affected by location, whereas age significantly affected both basal and GnRH-stimulated testosterone secretion and seminal quality (Serengeti Plains only) in sexually mature males. The striking seminal/endocrine differences among pride (breeding) males of different ages raises questions about the impact of age on individual reproductive performance in this species.

The extent and progression of exposure to feline infectious peritonitis (FIP) virus in the cheetah, Acinonyx jubatus, was monitored by a world-wide serological survey with indirect fluorescent antibody titers to coronavirus. The indirect fluorescent antibody assay was validated by Western blots, which showed that all indirect fluorescent antibody-positive cheetah sera detected both domestic cat and cheetah coronavirus structural proteins. There was a poor correlation between indirect fluorescent antibody results and the presence of coronaviruslike particles in cheetah feces, suggesting that electron microscopic detection of shed particles may not be an easily interpreted diagnostic parameter for FIP disease. Low, but verifiable (by Western blots [immunoblots]) antibody titers against coronavirus were detected in eight free-ranging cheetahs from east Africa as well as from captive cheetahs throughout the world. Of 20 North American cheetah facilities screened, 9 had cheetahs with measurable antibodies to feline coronavirus. Five facilities showed patterns of an ongoing epizootic. Retrospective FIP virus titers of an FIP outbreak in a cheetah-breeding facility in Oregon were monitored over a 5-year period and are interpreted here in terms of clinical disease progression. During that outbreak the morbidity was over 90% and the mortality was 60%, far greater than any previously reported epizootic of FIP in any cat species. Age of infection was a significant risk factor in this epizootic, with infants (less than 3 months old) displaying significantly higher risk for mortality than subadults or adults. Based upon these observations, empirical generalizations are drawn which address epidemiologic concerns for cheetahs in the context of this lethal infectious agent.

Five species of felids (cheetah, North Chinese leopard, tiger, lion, and puma) were serially injected with a source of follicle-stimulating hormone (FSH) to evaluate its effect on ovarian activity. Animals were subjected to laparoscopy before and after gonadotropin treatment, and the number and appearance of mature ovarian follicles (MF), corpora hemorrhagica (CH), and corpora lutea (CL) were recorded and photographed. Ovarian morphologic features, including MF, CH, and CL anatomy, were similar among various species. A dosage of 10 mg of FSH given IM daily for 3 to 5 days was effective in stimulating ovulation in 8 of 8 cheetahs. The same dosage for 5 days stimulated ovulation in 6 of 7 North Chinese leopards, 4 of 7 tigers, 2 lions, and 1 puma. An injection of a luteinizing hormone source was not required to induce follicular rupture. The number of ovulations eventually obtained appeared related to the activity prior to FSH treatment. Females with evidence of follicular development prior to treatment eventually produced a combined average (+/- SEM) of 5.2 +/- 1.3 CH and CL, in comparison with 2.3 +/- 0.8 CH and CL observed in cats with no identifiable follicles before FSH injection. The species varied in the number of ovulation sites (combined number of CH and CL) after treatment, with the cheetahs, leopards, tigers, lions, and puma producing a mean of 5.5 +/- 0.9, 1.5 +/- 0.2, 4.7 +/- 3.1, 1.5 +/- 0.5, and 3.0 +/- 0.0, respectively. The cheetahs, North Chinese leopards, and tigers failed to develop a refractory response to the gonadotropin, as certain females receiving FSH twice at a 1-year interval produced a similar ovulatory response.

The influence of electroejaculation on the acute response in serum cortisol, testosterone and luteinizing hormone (LH) was studied in the South African cheetah . Males were either anesthetized with CT-1341 and 1) serially bled only (controls, n = 7) or 2) serially bled during and following a regimented protocol of rectal probe electroejaculation (n = 14). In the control cheetahs , mean cortisol concentrations declined over time (P less than 0.05) and neither testosterone nor LH varied over the 145-min sampling interval. Serum cortisol rose immediately in electroejaculated cheetahs , peaked at the end of electroejaculation in 13 of 14 males and then declined during the next 90 min. Temporal profiles and serum levels of testosterone and LH were similar in the electroejaculated and control groups (P greater than 0.05). Within individual cheetahs , serum levels of LH and testosterone were highly correlated (r = 0.77, P less than 0.01). Awake (n = 2) and CT-1341 anesthetized (n = 2) cheetahs also were bled and then challenged with an i.m. injection of 25 IU adrenocorticotropic hormone (ACTH, Cortrosyn). Serial blood samples were collected during the next 2 h and assayed. Cortisol concentrations prior to ACTH administration were greater in awake than in anesthetized males. In all animals, cortisol rose immediately and peaked within 30-60 min of injection. Whereas all 4 ACTH-treated cheetahs produced cortisol titers in excess of 200 ng/ml, only 4 of 14 electroejaculated males produced cortisol levels comparable to this concentration range. Neither testosterone nor LH profiles were affected by ACTH-induced elevations in cortisol.(ABSTRACT TRUNCATED AT 250 WORDS)

The banded chromosomes of the giant panda (Ailuropoda melanoleuca) and a hybrid bear (Ursus middendorfi X Thalarctos maritimus) have been studied and compared to those of other carnivores. The giant panda shares only a few apparently homologous chromosome arms with the bear (Ursidae) and equally few with the lesser panda, Ailurus fulgens (Procyonidae). Chromosomal evidence suggests appropriate placement of the giant panda in a family separate from the Uridae and the Procyonidae.

A renal calculus composed principally of the amino acid, cystine, was found in an 8-year-old male maned wolf (Chrysocyon brachyurus). Cystine crystals were found in the urine sediment. The renal clearance of 10 amino acids was abnormal, whereas reabsorption of others was normal. The renal clearance of cystine, lysine, ornithine, and arginine exceeded the filtered load. The renal tubular handling of glucose, phosphate, sodium, potassium, and uric acid was identical to that for the clinically normal dog. These findings indicated an isolated renal tubular defect for cystine and other amino acids.

The efficacy of an inactivated vaccine for the prevention of feline viral rhinotracheitis (FVR), feline caliciviral disease (FCVD), and feline panleukopenia (FPL) was tested in 27 nondomestic adult felids from 7 species. The vaccine was given IM at the standard domestic cat dose in 19 animals and double this dose in 8 others. The animals were vaccinated either 1, 2, or 3 times. Serum-neutralization (SN) antibodies to FVR (mean SN titer, 23) developed in all 15 animals that were previously seronegative, and SN antibodies to FCVD (mean SN titer, 11) developed in 19 of 21 animals that were previously seronegative. There was no significant increase of SN antibody titers by doubling the vaccine dose or by administering a 3rd vaccination. The optimal response could be obtained by using the domestic cat vaccination protocol of a single dose given twice, 4 weeks apart. The critical evaluation of the SN antibody titer for FPL was complicated by preexisting titers to FPL from previous vaccinations, but in 23 animals the titers became higher, whereas they remained unchanged in only 4 animals. The persistence of the SN titers was evaluated 7 to 9 months later and found to be satisfactory for FVR (mean SN titer, 18) FCVD (mean SN titers, 43) and FPL (mean SN titer, 517). Enhanced persistence of titer could not be demonstrated by doubling the dose or administering a 3rd vaccination.

Five domestic ferrets previously maintained for 12 weeks under a 16L:8D photoperiod were electroejaculated weekly for 15-65 weeks while continuing to be exposed to the prolonged light cycle. Two ferrets sustained spermatogenesis for 20 and 26 weeks, while sperm production in the remaining males either was sporadic or decreased, remained depressed and then increased to peak levels observed in other males. Regardless of the temporal spermatogenesis patterns within males, the number of electroejaculated spermatozoa with residual cytoplasmic droplets or abnormal acrosomes increased in all ferrets over time. Diluted ejaculates meeting artificial insemination criteria were deposited intravaginally or by transabdominal laparoscopy into the uterine horns of females treated 0 or 24 h earlier with 90 i.u. hCG. Vaginal insemination was ineffective (0 pregnancies in 10 attempts), but 17/24 ferrets (70.8%) inseminated laparoscopically became pregnant and delivered live young (mean litter size, 5.2 kits). Number of motile spermatozoa deposited in utero (1.6-10.0 x 10(6) cells), presence of glycerol in the sperm dilution medium (0 versus 4%) and time of hCG administration (0 versus 24 h before insemination) had no effect on pregnancy results or litter size.

Two types of killed canine distemper virus (CDV) vaccine and a modified-live CDV vaccine were clinically evaluated in four species of exotic carnivores. In 16 trials in which 13 red pandas (Ailurus fulgens) were given the killed vaccine, only 1 animal had a virus-neutralization titer that exceeded 1:100. A red panda given modified-live CDV vaccine deemed safe for gray foxes and ferrets died of bacterial pneumonia 16 days later. There was no pathologic evidence of canine distemper in that panda. The same modified-live vaccine proved to be immunogenic and safe in 12 bush dogs (Speothos venaticus), 5 maned wolves (Chrysocyon brachyurus), and 3 fennec foxes (Fennecus zerda) in which virus-neutralization titers often exceeded 1:512 and persisted for several months after vaccination.

Canine parvovirus (CPV) infections occurred in 5 of 35 South American canids at the Department of Conservation (DC), a breeding facility of the National Zoological Park in Front Royal, Va. The clinical signs were anorexia, lethargy, diarrhea, and vomiting. Three of the affected canids survived and had high hemagglutination-inhibition titers to CPV in the recovery period. Necropsy of the 2 that died revealed extensive necrosis of the intestinal mucosa; CPV particles were observed by electron microscopy in the intestinal contents of both animals. Six of the 30 canids that remained healthy had high hemagglutination-inhibition titers to CPV prior to the episode of illness, indicating earlier subclinical exposure. Pet dogs belonging to DC personnel that were screened as a possible source of the infection had no evidence of disease. All canids (including pet dogs) on the DC grounds were vaccinated repeatedly with a killed feline panleukopenia virus product after the episode, with little or no effect on existing titers.

A circannual analysis was made of serum cortisol, luteinizing hormone (LH), and testosterone concentrations in the male clouded leopard (Neofelis nebulosa). Group I males (n = 4), maintained in a standardized environment, were bled serially during a regimented anesthesia/electroejaculation episode occurring monthly (beginning in January, ending in December). Additional sampling intervals were conducted under anesthesia only (control, n = 8), anesthesia plus a single adrenocorticotropin hormone challenge (ACTH, Cortrosyn, n = 4), or anesthesia plus a single 25 micrograms injection of gonadotropin-releasing hormone (GnRH, Gonadorelin, n = 4). Group II males (n = 6) from various zoological collections were sampled serially under the same semen collection conditions on one random occasion within the year. Serum cortisol levels were 2 times greater than values measured in comparable studies involving other felid species. Cortisol concentrations were similar during electroejaculation and control (anesthesia only) episodes, and mean levels did not rise as a result of semen collection. Adrenocorticotropin caused an immediate rise in cortisol to levels at least 1.5 times greater than electroejaculated or control counterparts. Mean concentrations of basal cortisol in individual males gradually increased as the year progressed, possibly as a consequence of repeated psychogenic stress. Between seasons, there were no differences in mean LH; however, testosterone levels were greater (p less than 0.05) in the winter compared to all other seasons. There were no differences (p greater than 0.05) between individual males in secretory patterns or mean concentrations of cortisol, LH, or testosterone. Within males, distinct temporal fluctuations were observed in both LH and testosterone during the approximately 80-min sampling interval. Neither LH nor testosterone profiles appeared affected by cortisol patterns during electroejaculation or after an ACTH challenge. A bolus of GnRH induced a marked rise in serum LH and testosterone within 15 and 30 min respectively, indicating that these two hormones were coupled. Both LH and testosterone profiles in Group II males mimicked those in Group I; concentrations of cortisol in Group II males immobilized on one occasion were similar to those of Group I animals sampled from January-May but appeared to be less than values measured from June-December. These data demonstrate that the clouded leopard, compared to other felids, produces markedly elevated concentrations of cortisol, which are likely related to an aggressive behavioral temperament.(ABSTRACT TRUNCATED AT 400 WORDS)

A fatal disease occurred in a lesser panda (Ailurus fulgens) 2 weeks after vaccination with modified live distemper vaccine. The disease clinically resembled canine distemper. Pathologically there was giant cell pneumonia, with canine distemper viral inclusion bodies in pulmonary and digestive tract epithelium. Viral isolates were indicative of an attenuated strain rather than virulent types.

Nineteen bears, representing five species (Helarctos malayanus, Ursus americanus, Ursus arctos, Tremarctos ornatus, Melursus ursinus) were immobilized a total of 52 times using either phencyclidine-promazine or tiletamine-zolazepam. Blood gas, hematology, and serum biochemistry values were determined during the immobilizations. Immobilizations conducted with tiletamine-zolazepam were characterized by rapid induction and recovery times, good muscle relaxation, and relative freedom from convulsions. Bears immobilized with phencyclidine-promazine had longer induction and recovery times and showed convulsive activity in 29% of the trials with that combination.

Ejaculates were analyzed from clouded leopards (Neofelis nebulosa) subjected to a regimented anesthesia/electroejaculation protocol. Group I males (n = 4), maintained individually in an environment with natural fluctuations in photoperiod, were electroejaculated on the same day at monthly intervals (January-December). Group II clouded leopards (n = 8), maintained in random zoo populations throughout the U.S., were evaluated on a single occasion. Phase contrast and electron microscopy indicated a high proportion of structurally abnormal spermatozoa in seminal fluid (Group I range, 14.8-78.9%; Group II range, 32.3-93.0%), the predominant deformity being a tightly coiled flagellum. Semen quality, including spermatozoal concentration and the incidence of abnormal sperm forms, varied (p less than 0.05) among males. Evaluating the numbers of motile spermatozoa/ejaculate (MS/E) among individual males from Group I on a monthly basis suggested a seasonal influence; gradually increasing MS/E values with peaks in June and July were observed in three of four animals. A simultaneous analysis of international breeding records for captive female clouded leopards demonstrated that 46.2% of parturitions occurred in March and April, indicating that most estrual periods occurred from late December through February. These data suggest that a physiological asymmetry may exist in peak reproductive performance between the male and female clouded leopard, perhaps as a result of differing adaptations to the captive environment. Motile spermatozoa can be recovered throughout the year using electroejaculation and, when used over time, a standardized procedure can determine a hierarchy of seminal quality among males of unknown reproductive potential. The relatively high proportions of structurally abnormal spermatozoa in the ejaculates of the clouded leopard may be related to a low degree of genetic variation within the species and/or hyperadrenal activity in captive populations.

Ejaculate-endocrine characteristics were measured in 23 captive cheetahs (Acinonyx jubatus jubatus) in North American zoos and in 8 free-ranging cheetahs (A.j. raineyi) in eastern Africa (Tanzania). A standardized electroejaculation protocol was used, and numbers of motile spermatozoa were similar (p greater than 0.05) between groups. Of the spermatozoa collected by electroejaculation, 70.6 +/- 3.3% and 75.9 +/- 4.4% were morphologically abnormal in the captive "North American" and in the free-ranging, eastern African populations, respectively. Adrenal activity, as measured by an acute, temporal rise and fall in serum cortisol levels during and after electroejaculation, was no different (p greater than 0.05) between groups. Although serum luteinizing hormone (LH) levels were less (p less than 0.05) in the free-ranging than in the captive animals, serum testosterone concentrations were similar. The data indicate that the comparatively poor reproductive performance of cheetahs maintained in zoological parks is not attributable to a captivity-induced response afflicting the male. Furthermore, there is no evidence that ejaculate/endocrine characteristics differ between the two subspecies. Because adrenal/gonadal activity and the number of pleiomorphic spermatozoa are similar between the test groups, the results suggest that spermatozoal diversity originates as a result of the extreme genetic monomorphism observed universally in the species.

Luteinizing hormone releasing hormone (LHRH, 50 micrograms) or saline was administered (i.m.) to adult female and male cheetahs under anaesthesia to evaluate pituitary and gonadal response. Serum LH levels did not fluctuate over a 120-min sampling period in saline-treated animals. Serum LH concentrations were raised (P less than 0.05) in both female and male cheetahs after LHRH injection, the temporal response being similar to previously reported results in unanaesthetized, domestic carnivores. The magnitude of the LHRH-induced LH response was sex-dependent. Over a 120-min post-injection period both saline control and LHRH-induced LH levels were about twofold greater in males than females. Although LHRH had no acute influence on ovarian oestradiol-17 beta production in the female, serum testosterone levels were raised (P less than 0.05) in male cheetahs by 60 min after treatment. This study (1) provides introductory endocrine information on the cheetah, an endangered species, and (2) indicates that exogenous LHRH is effective in acutely altering pituitary (female) and pituitary/gonadal (male) function in an anaesthetized, non-domestic felid.

Fifty-eight immobilizations were conducted using 21 lions (Panthera leo) and 4 drug combinations. The combinations used were ketamine-phencyclidine-promazine, xylazine-phencyclidine-promazine, xylazine-ketamine-phencyclidine-promazine, and tiletamine-zolazepam.

A population genetic survey of over 200 structural loci previously revealed that the South African cheetah (Acinonyx jubatus jubatus) has an extreme paucity of genetic variability, probably as a consequence of a severe population bottleneck in its recent past. The genetic monomorphism of the species is here extended to the major histocompatibility complex, since 14 reciprocal skin grafts between unrelated cheetahs were accepted. The apparent consequences of such genetic uniformity to the species include (i) great difficulty in captive breeding, (ii) a high degree of juvenile mortality in captivity and in the wild, and (iii) a high frequency of spermatozoal abnormalities in ejaculates. The species vulnerability of the cheetah was demonstrated by an epizootic of coronavirus-associated feline infectious peritonitis in an Oregon breeding colony in 1983. Exposure and spread of the coronavirus, which has a very low morbidity in domestic cats (approximately 1 percent), has decimated a heretofore productive and healthy captive population. The extreme genetic monomorphism, especially at the major histocompatibility complex, and the apparent hypersensitivity of the cheetah to a viral pathogen may be related, and provide a biological basis for understanding the adaptive significance of abundant genetic variation in outbred mammalian species.

Urinary concentrations of conjugated oestrone and pregnanediol-3-glucuronide were measured during and after spontaneous and induced oestrus and during pregnancy. Behavioural oestrus was preceded by a rise in oestrone values from less than 10 ng/mg creatinine (Cr) to peaks of 45 ng/mg Cr. Maximal lordotic response and mating activity coincided with the decline in oestrone levels. After presumed ovulation, urinary pregnanediol glucuronide concentrations increased from less than 5 to 15-30 ng/mg Cr. Further increases in this steroid (to 60-80 ng/mg Cr) occurred 114 days after mating, presumably coincident with implantation. These high levels of pregnanediol glucuronide were maintained for 3 weeks, began to decline 1 week before parturition and fell to a nadir (less than 5 ng/mg Cr) immediately after delivery. When FSH was administered i.m. for 5 days, urinary oestrone values rose markedly and were maximal (580 ng/mg Cr) on Day 7. Mating first occurred on Day 20 and 500 i.u. hCG were given i.m. Urinary pregnanediol glucuronide levels during the next 5 months were similar to those in the previous year during pregnancy with values rising 105-108 days after mating. However, no birth occurred. These results support the suggestion that pandas exhibit delayed implantation and demonstrate that the panda is responsive to exogenous gonadotrophins.

Although it is generally agreed that the giant panda (Ailuropoda melanoleuca) is a member of the order Carnivora, there has long been disagreement over whether it should be classified with bears, raccoons or as a single member of its own family. Four independent molecular and genetic measures lead to a consensus phylogeny for the giant and lesser pandas. The lesser panda diverged from New World procyonids at approximately the same time as their departure from ursids, while ancestors of the giant panda split from the ursid lineage much later, just before the radiation which led to modern bears. The giant panda's divergence was accompanied by a chromosomal reorganization which can be partially reconstructed from the ursid karyotype, but not from that of procyonids or the lesser panda. The apparently dramatic, but actually limited, distinctions between the giant panda and the bears in chromosomal and anatomical morphology provide a graphic mammalian example of the discordance of molecular and morphological (and chromosomal) evolutionary change.

Urinary excretion of oestrone conjugates, pregnanediol-3 alpha-glucuronide (PdG) and 20 alpha-hydroxypregn-4-en-3-one were measured from 8 weeks before oestrus to 2 weeks post partum and bioactive FSH was monitored during the periovulatory interval in a female giant panda. A biphasic urinary bioactive FSH excretory profile appeared to indicate a broad (approximately 10 day) follicular phase followed by a sharp preovulatory bioactive FSH surge coincident with an acute increase in urinary oestrone conjugates and behavioural oestrus. Weekly concentrations of urinary oestrone conjugates and PdG increased (P less than 0.001) by Week 9 of gestation with 20 alpha-hydroxypregn-4-en-3-one levels increasing 10-30-fold (P less than 0.001) between Weeks 11 and 14. These observations indicate that the monoestrous giant panda does not appear to require a prolonged period of endogenous FSH release or multiple FSH peaks for ovarian priming and follicle selection to proceed normally. Furthermore, the delayed rise in urinary steroid excretion during the second half of gestation in the giant panda supports the concept of delayed implantation while the estimation of steroid conjugates in urine offers a non-invasive approach for monitoring pregnancy status in this endangered species.

Of 42 maned wolves in zoos or live-trapped in Brazil, 34 had excessive cystine in their urine. Renal clearance studies of five of the affected wolves revealed a variable defect for the reabsorption of cystine and dibasic amino acids. The renal tubular handling of other solutes including glucose, phosphate, sodium, potassium, and uric acid was considered normal. Urinary calculi composed of cystine were found in four wolves and proved fatal in three of them. With the exception of the high incidence in this species, this hereditary disease resembles the disorder described in dogs and humans.

A combined population genetic and reproductive analysis was undertaken to compare free-ranging cheetahs from east Africa (Acinonyx jubatus raineyi) with the genetically impoverished and reproductively impaired south African subspecies (Acinonyx jubatus jubatus). Like that of their south African counterparts, the quality of semen specimens from east African cheetahs was poor, with a low concentration of spermatozoa (25.3 X 10(6) per ejaculate) and a high incidence of morphological abnormalities (79%). From an electrophoretic survey of the products of 49 genetic loci in A. jubatus raineyi, two allozyme polymorphisms were detected; one of these, for a nonspecific esterase, shows an allele that is rare (less than 1% incidence) in south African specimens. Estimates of polymorphism (2-4%) and average heterozygosity (0.0004-0.014) affirm the cheetah as the least genetically variable felid species. The genetic distance between south and east African cheetahs was low (0.004), suggesting that the development of genetic uniformity preceded the recent geographic isolation of the subspecies. We propose that at least two population bottlenecks followed by inbreeding produced the modern cheetah species. The first and most extreme was ancient, possibly late Pleistocene (circa 10,000 years ago); the second was more recent (within the last century) and led to the south African populations.

Semen was collected by a standardized electroejaculation procedure from a giant panda on 4 occasions. Ejaculate volume, sperm count and % sperm motility were 2.3-3.6 ml, 62-562 x 10(6) spermatozoa/ml and 45-85%, respectively. The results, although limited to a single male, suggested a seasonal influence on ejaculate and gonadal parameters with improved ejaculate volume, sperm motility and increased testicular size in the season proximate to the female's oestrous period. Frozen-thawed spermatozoa were motile with no apparent abnormalities induced by the freezing procedure.

Male cheetahs, tigers, leopards, and pumas maintained under the same conditions were anesthetized and 1) serially bled before, during, and after electroejaculation (EE); 2) serially bled only (AO); or 3) serially bled before and after receiving adrenocorticotropin hormone (ACTH). Ejaculates from leopards contained higher (p less than 0.05) sperm concentrations than cheetahs and pumas but lower (p less than 0.05) sperm motility ratings than all other species. Tigers produced a larger seminal volume and the greatest number of motile sperm/ejaculate (p less than 0.05). The percentage of morphologically abnormal spermatozoa was greater (p less than 0.05) in cheetahs (64.6%), leopards (79.5%), and pumas (73.5%) than in tigers (37.5%). The most prevalent spermatozoal deformities included a tightly coiled or bent flagellum, a deranged midpiece, or a residual cytoplasmic droplet. Mean baseline serum cortisol concentrations in leopards were 2- and 4-fold greater (p less than 0.05) than in tigers and cheetahs, respectively. Basal cortisol concentrations in pumas were similar to those of tigers, but irrespective of treatment increased 2-fold (p less than 0.01) during the bleeding period. An acute rise and fall in cortisol attributable to EE was observed only in cheetahs. In tigers and leopards, mean peak cortisol concentrations after ACTH were similar to maximal values observed after EE. However, peak cortisol levels in cheetahs and pumas after ACTH were greater (p less than 0.01) than the concentrations measured after EE, indicating that these manipulatory procedures were not eliciting a maximal adrenal response. In the EE groups, luteinizing hormone (LH) and testosterone levels in cheetahs were lower (p less than 0.05) than in other species, whereas levels of both hormones were comparable (p greater than 0.05) in tigers, leopards, and pumas. Elevated cortisol levels in cheetahs and pumas had no discernible effect on LH/testosterone patterns; however, the results were equivocal in tigers, and, among leopards, testosterone concentrations consistently declined over time. In this study, using a standardized approach, we identify different ejaculate and endocrine characteristics of captive cheetahs, tigers, leopards, and pumas. The data extend earlier observations and demonstrate that some, but not all, Felidae species ejaculate high numbers of pleiomorphic spermatozoa. However, inter-species differences in sperm integrity do not appear related to inter-species variations in cortisol, LH, or testosterone. The observation of continuously declining testosterone concentrations only in leopards after AO, EE, or ACTH treatment suggests that rising and/or elevated cortiso

Analysis of 40 semen samples collected by electroejaculation from 18 cheetahs revealed no major differences in seminal traits among Transvaal, South West (Namibia) or hybrid (Transvaal X South West) males. However, mean spermatozoal concentration (14.5 X 10(6) spermatozoa/ml of ejaculate) and percent motility (54.0%) were less in cheetahs than in domestic cats (147.0 X 10(6) spermatozoa/ml of ejaculate, 77.0% motility) subjected to the same electroejaculation regimen. On the average, cheetah ejaculates contained 71.0% morphologically abnormal spermatozoa compared to 29.1% aberrant spermatozoal forms in the domestic cat. These results indicate that seminal characteristics in the cheetah are markedly inferior compared to the domestic cat, particularly with respect to the incidence of pleiomorphic spermatozoa. Because a recent parallel study demonstrates that the cheetah lacks genetic variation, it appears likely that spermatozoal abnormalities are a genetic consequence of genomic homozygosity characteristic of this endangered species.