Clinical and pathological findings of a newly recognized disease of elephants caused by endotheliotropic herpesviruses.

The unique clinical and pathological findings in nine Asian (Elephas maximus) and two African (Loxodonta africana) elephants from North American Zoos with a highly fatal disease caused by novel endotheliotropic herpesviruses are described. Identification of the viruses by molecular techniques and some epidemiological aspects of the disease were previously reported. Consensus primer polymerase chain reaction (PCR) combined with sequencing yielded molecular evidence that confirmed the presence of two novel but related herpesviruses associated with the disease, one in Asian elephants and the second in African elephants. Disease onset was acute, with lethargy, edema of the head and thoracic limbs, oral ulceration and cyanosis of the tongue followed by death of most animals in 1 to 7 days. Pertinent laboratory findings in two of three clinically evaluated animals included lymphocytopenia and thrombocytopenia. Two affected young Asian elephants recovered after a 3 to 4 wk course of therapy with the anti-herpesvirus drug famciclovir. Necropsy findings in the fatal cases included pericardial effusion and extensive petechial hemorrhages in the heart and throughout the peritoneal cavity, hepatomegaly, cyanosis of the tongue, intestinal hemorrhage, and ulceration. Histologically, there were extensive microhemorrhages and edema throughout the myocardium and mild, subacute myocarditis. Similar hemorrhagic lesions with inflammation were evident in the tongue, liver, and large intestine. Lesions in these target organs were accompanied by amphophilic to basophilic intranuclear viral inclusion bodies in capillary endothelial cells. Transmission electron microscopy of the endothelial inclusion bodies revealed 80 to 92 nm diameter viral capsids consistent with herpesvirus morphology. The short course of the herpesvirus infections, with sudden deaths in all but the two surviving elephants, was ascribed to acute cardiac failure attributed to herpesvirus-induced capillary injury with extensive myocardial hemorrhage and edema.

Disparate phylogeographic patterns of molecular genetic variation in four closely related South American small cat species.

Tissue specimens from four species of Neotropical small cats (Oncifelis geoffroyi, N = 38; O. guigna, N = 6; Leopardus tigrinus, N = 32; Lynchailurus colocolo, N = 22) collected from throughout their distribution were examined for patterns of DNA sequence variation using three mitochondrial genes, 16S rRNA, ATP8, and NADH-5. Patterns between and among O. guigna and O. geoffroyi individuals were assessed further from size variation at 20 microsatellite loci. Phylogenetic analyses using mitochondrial DNA sequences revealed monophyletic clustering of the four species, plus evidence of natural hybridization between L. tigrinus and L. colocolo in areas of range overlap and discrete population subdivisions reflecting geographical isolation. Several commonly accepted subspecies partitions were affirmed for L. colocolo, but not for O. geoffroyi. The lack of geographical substructure in O. geoffroyi was recapitulated with the microsatellite data, as was the monophyletic clustering of O. guigna and O. geoffroyi individuals. L. tigrinus forms two phylogeographic clusters which correspond to L.t. oncilla (from Costa Rica) and L.t. guttula (from Brazil) and which have mitochondrial DNA (mtDNA) genetic distance estimates comparable to interspecific values between other ocelot lineage species. Using feline-specific calibration rates for mitochondrial DNA mutation rates, we estimated that extant lineages of O. guigna diverged 0.4 million years ago (Ma), compared with 1.7 Ma for L. colocolo, 2.0 Ma for O. geoffroyi, and 3.7 Ma for L. tigrinus.

Special challenges of maintaining wild animals in captivity in North America.

The maintenance of wild animals in captivity in North America is regulated by a number of different laws and government agencies in each country. Member institutions of zoo and aquarium associations in Canada, the United States of America and Mexico experience an extra tier of regulation in the form of industry standards, which are sometimes stricter than those imposed by government. Climate, natural disasters and harmful pest species all contribute to the challenge of keeping animals in certain locales. Vigilance against zoonotic disease transmission is maintained through industry and government-mandated sanitation standards, which are fortified by reporting regulations of local, regional and Federal health agencies. Current controversies in the keeping of particular taxa in North America include the threat to non-human primate breeding programmes precipitated by strict new import regulations, the fear of herpesvirus B infection, and commercial airline transport bans. Successive human fatalities among elephant handlers have prompted the industry and governments to re-examine the manner in which these potentially dangerous creatures are maintained in captivity.

Cardiopulmonary effects of three prolonged periods of isoflurane anesthesia in an adult elephant.

An adult 3,500-kg female African elephant was anesthetized 3 times for treatment of subcutaneous fistulas over the lateral aspect of each cubitus (anesthesia 1 and 2) and for repair of a fractured tusk (anesthesia 3). Lateral recumbency and anesthesia were achieved with etorphine (anesthesia 1 and 2) or etorphine and azaperone (anesthesia 3). The elephant's trachea was intubated and anesthesia was maintained by use of isoflurane and oxygen delivered via 2 standard large animal anesthesia machines joined in parallel. The range of total recumbency time was 2.4 to 3.3 hours. Breathing and heart rates, systemic arterial pressure, rectal temperature, PaO2, pH, and end-tidal gases were monitored. After administration of etorphine, measurements were made while the air-breathing elephant was recumbent, then every 5 minutes (cardiovascular) or 15 minutes (blood gases) after commencement of administration of isoflurane and oxygen. Tachycardia and hypertension were detected after administration of etorphine, but heart rate and systemic arterial pressure decreased to within normal ranges after administration of isoflurane and oxygen. The elephant remained well oxygenated while anesthetized and breathing a high oxygen mixture. The elephant had an uneventful recovery from each anesthesia.

Freemartinism in a captive herd of Rocky Mountain bighorn sheep (Ovis canadensis).

Freemartinism in two animals from a captive herd of Rocky Mountain bighorn sheep (Ovis canadensis) at the Denver Zoological Gardens (Denver, Colorado, USA) is described. A young ewe had female external genitalia, a masculine appearance, and demonstrated male behavior as she matured. Another ewe with female external genitalia died as a yearling. Necropsy revealed a non-patent vagina and internal male genitalia. Both females were chimeric with karyotypes containing XX and XY sex chromosomes.

Laparoscopic evaluation of the reproductive organs and abdominal cavity content of the lowland gorilla.

Laparoscopy was used to examine the abdominal cavity and evaluate reproductive competence in six adult female gorillas which had not demonstrated copulatory activity within 12 months of examination. Blood samples were obtained on the day of laparoscopy and analyzed for serum estradiol-17ß and progesterone. The uterus and oviducts of each gorilla appeared morphologically normal and free of lesions. The ovaries of three animals contained anatomical evidence of activity as indicated by the presence of luteal scars and a prominent corpus hemorrhagicum, corpus luteum, or developing follicles. No visible luteal or follicular tissue was observed on the ovaries of three gorillas. In the eldest female (age, 22 years) the ovaries were flaccid in texture and demonstrated irregularities in ovarian surface integrity, similar to that observed in women during reproductive senescence. Serum concentrations of estradiol-17ß and progesterone on the day of laparoscopy corresponded with observed ovarian activity. All six females had extensive adhesions in the abdominal cavity exclusive of the reproductive organs. Numerous gut, omentum, and liver to peritoneum adhesions were observed. The results indicated that laparoscopy was a safe and effective method for evaluating anatomical competence in the gorilla. Uterine and ovarian integrity appeared morphologically normal, but only three of the six gorillas showed evidence of ongoing ovarian activity.

Isolation of Campylobacter fetus subsp jejuni from zoo animals.

Over a 1-year period, 619 fecal specimens from animals at the Denver Zoo were cultured for Campylobacter fetus subsp jejuni. The organism was isolated from 35 animals, including 12 primates, 2 felids, a red panda, 13 hooved animals, 6 birds, and 1 reptile. Of 44 cultured fecal specimens from diarrheal animals, 31.8% were positive for Campylobacter, whereas only 5.6% of 575 specimens from animals without diarrhea were positive (P less than 0.001). Among 25 isolates tested, 12 serotypes were represented; several of these serotypes are commonly associated with Campylobacter enteritis in human beings. Campylobacter fetus subsp jejuni was isolated from 8% of 75 wild pigeons trapped on the zoo premises during winter months and from 26% of 75 trapped during March and April (P less than 0.01).

Fatal airsacculitis and pneumonia, with abortion, in an orangutan.

A 14-year-old, 76-kg female orangutan (Pongo pygmaeus) aborted a premature fetus and within hours experienced severe respiratory distress and died. Necropsy revealed evidence of chronic airsacculitis, acute bronchopneumonia, and terminal sepsis. Escherichia coli was isolated from cultures of heart blood, tracheal fluid, air-sac tissue and fluid, lung, and bile. Retrospectively, it was established that the animal had had a periodic nasal discharge for many months. The aborted infant had no evidence of infection.

Salmonellosis and arizonosis in the reptile collection at the National Zoological Park.

The prevalence of Salmonella and Arizona organisms in the reptile collection at the National Zoological Park was investigated. Culture of specimens from 311 reptiles, while live or at necropsy, yielded yielded 117 positive results, for an overall infection rate of 37%. Snakes had the highest rate, 55% (69 of 125); lizards had an intermediate rate, 36% (46 of 129); and turtles and tortoises had the lowest rate, 3% (2 of 63). Twenty-four serotypes of Salmonella enteritidis, 1 of S choleraesuis, and 39 of Arizona hinshawii were represented. While clinical illness was never directly attributed to infection with these organisms, pure cultures of Salmonella and Arizona were recovered at necropsy from some reptiles with gross and/or histologic lesions in the gastrointestinal tract, liver, spleen, and blood vessels. However, numerous other concurrent diseases and management problems were often considered the immediate cause of death, with Salmonella and Arizona being ready and significant opportunistic pathogens contributing to the demise of the reptiles.