FREE-RANGING OCELOTS (LEOPARDUS PARDALIS): HEMATOLOGY AND SERUM CHEMISTRY REFERENCE VALUES.

Acquiring baseline physiologic data for animals from a free-ranging wildlife species is an elusive objective. Between 1990 and 2020, a monitoring program on the last population of ocelot (Leopardus pardalis) to inhabit public land in the United States yielded 139 blood samples from 67 individual animals. Ocelots were live trapped and anesthetized for census and radiotelemetric studies. The protocol included morphometrics, photographs, electronic identification, and blood collection. Complete blood count and serum chemistry were performed, and after sorting of the data to remove unhealthy individuals and occasional outliers, the dataset provided sufficient information to compute reliable reference intervals (RI). According to the American Society of Veterinary Clinical Pathology consensus guidelines, RI should be elaborated by using data from each reference individual only once. RI by random selection was determined when several measurements were available over time from one same animal. Second, RI were also computed allowing repeat measurements for reference individuals, exclusively to characterize and quantify the effect on the data distribution and on the generated RI. A summary of published RI for various species of wild felids is also presented. The variations observed between species is due not only to species differences but also to variation in measurement methods and RI study design. Overall, accurate blood work interpretation requires RI generated from a healthy population, with defined measurement methods and state-of-the-art RI study design. Of note, calcium is typically tightly regulated in all mammals, as illustrated by the narrow RI (8.5-10.8 mg/dl); conversely, finding a narrow RI in calcium across as many as 49 healthy individuals suggests a high-quality design study.

THREE CASES OF ACUTE BACTERIAL SEPSIS IN PYGMY HIPPOPOTAMUS (CHOEROPSIS LIBERIENSIS) CALVE SIBLINGS.

A multiparous pygmy hippopotamus (Choeropsis liberiensis) dam produced three consecutive calves that died acutely at 13-15 wk of age from bacterial sepsis, for which diagnostic and therapeutic intervention was not possible. Streptococcus iniae (Cases 1 and 3), Escherichia coli (Case 2), and an unidentified member of the family Pasteurellaceae (Case 1) were identified in postmortem tissues through bacterial culture followed by standard and molecular identification methods. After the loss of two calves, a series of vaccinations were administered to the dam during the third pregnancy to enhance transplacental and colostral transfer of antibodies to the calf. The third calf did not survive, and the source of the bacterial infection in these three calves was undetermined. Prior to and after the birth of the fourth calf, nutritional and nutraceutical supplements were provided to the dam and calf. Additionally, pest control around the barn was enhanced. The fourth calf survived. Pygmy hippopotamus calves at the age of 13-15 wk may have increased susceptibility to bacterial infection, possibly due to waning maternally derived immunity. The findings in these cases, combined with a previous association of S. iniae in pygmy hippopotamus deaths, suggest that this bacterium is an especially important pathogen of the endangered pygmy hippopotamus.

Trypanosoma cruzi and Incidental Sarcocystis spp. in Endangered Ocelots (Leopardus pardalis) of South Texas, USA.

The federally endangered ocelot (Leopardus pardalis) population of south Texas, USA is declining; fewer than an estimated 80 ocelots remain. South Texas has robust transmission of Trypanosoma cruzi, the protozoan parasite causing Chagas disease in humans and various mammals. This parasite's impact in ocelots is unknown. Blood from live-trapped ocelots was collected by US Fish and Wildlife Service personnel in an annual monitoring program; additionally, tissues were obtained from carcasses collected from 2010 to 2017 around Laguna Atascosa National Wildlife Refuge in south Texas and placed in scientific collections. Variable samples were available from 21 ocelots: skeletal muscle (n=15), heart tissue (n=5), lung (n=1), kidney (n=1), spleen (n=1), liver (n=1), blood clot (n=9), and serum (n=3). Overall, 3/21 (14.3%) ocelots showed evidence of T. cruzi infection or exposure, with T. cruzi PCR-positive samples of skeletal muscle, heart, and blood clot, respectively. All three were infected with the T. cruzi discrete taxonomic unit "TcI"; one of these ocelots also had anti-T. cruzi antibodies. Lymphoplasmacytic inflammation was noted in the PCR-positive heart tissue and in some PCR-negative tissues from this and other individuals. Incidentally, Sarcocystis spp. were noted histologically in five ocelots. Trypanosoma cruzi infection and associated cardiac lesions suggest that this parasite should be further investigated in vulnerable populations.

Transcriptomic Profiling of Fibropapillomatosis in Green Sea Turtles (Chelonia mydas) From South Texas.

Sea turtle fibropapillomatosis (FP) is a tumor promoting disease that is one of several threats globally to endangered sea turtle populations. The prevalence of FP is highest in green sea turtle (Chelonia mydas) populations, and historically has shown considerable temporal growth. FP tumors can significantly affect the ability of turtles to forage for food and avoid predation and can grow to debilitating sizes. In the current study, based in South Texas, we have applied transcriptome sequencing to FP tumors and healthy control tissue to study the gene expression profiles of FP. By identifying differentially expressed turtle genes in FP, and matching these genes to their closest human ortholog we draw on the wealth of human based knowledge, specifically human cancer, to identify new insights into the biology of sea turtle FP. We show that several genes aberrantly expressed in FP tumors have known tumor promoting biology in humans, including CTHRC1 and NLRC5, and provide support that disruption of the Wnt signaling pathway is a feature of FP. Further, we profiled the expression of current targets of immune checkpoint inhibitors from human oncology in FP tumors and identified potential candidates for future studies.

Molecular characterization of a marine turtle tumor epizootic, profiling external, internal and postsurgical regrowth tumors.

Sea turtle populations are under threat from an epizootic tumor disease (animal epidemic) known as fibropapillomatosis. Fibropapillomatosis continues to spread geographically, with prevalence of the disease also growing at many longer-affected sites globally. However, we do not yet understand the precise environmental, mutational and viral events driving fibropapillomatosis tumor formation and progression.Here we perform transcriptomic and immunohistochemical profiling of five fibropapillomatosis tumor types: external new, established and postsurgical regrowth tumors, and internal lung and kidney tumors. We reveal that internal tumors are molecularly distinct from the more common external tumors. However, they have a small number of conserved potentially therapeutically targetable molecular vulnerabilities in common, such as the MAPK, Wnt, TGFß and TNF oncogenic signaling pathways. These conserved oncogenic drivers recapitulate remarkably well the core pan-cancer drivers responsible for human cancers. Fibropapillomatosis has been considered benign, but metastatic-related transcriptional signatures are strongly activated in kidney and established external tumors. Tumors in turtles with poor outcomes (died/euthanized) have genes associated with apoptosis and immune function suppressed, with these genes providing putative predictive biomarkers.Together, these results offer an improved understanding of fibropapillomatosis tumorigenesis and provide insights into the origins, inter-tumor relationships, and therapeutic treatment for this wildlife epizootic.

COMPLICATIONS ASSOCIATED WITH PREGNANCY AND PARTURITION IN AFRICAN WHITE-BELLIED PANGOLINS (PHATAGINUS TRICUSPIS).

There are no studies to date on the normal reproductive physiology of African white-bellied pangolins (Phataginus tricuspis). As a reclusive species, little is known about normal gestation, successful parturition, and potential complications during pregnancy. Ten female P. tricuspis were diagnosed as pregnant and monitored under professional care. Five developed complications during pregnancies or during parturition and are detailed in this case series. Dystocia occurred in two dams each, with malposition of a singleton fetus. Both dams were successfully treated with surgical intervention by caesarian section. Of the two individuals, one fetus was nonviable, but the other neonate survived and was reared by the dam to weaning. A third pregnant female died during pregnancy from septicemia resulting in death of the preterm fetus. The two additional dams of the five gave birth to full-term neonates. One fetus was stillborn with evidence of fetal distress, and the other died immediately after birth (perinatal death) with undetermined etiology. Based on this case series, complications associated with pregnancy occur in P. tricuspis, indicating the need for further study and close monitoring during impending parturition.

Goats are a potential reservoir for the herpesvirus (MCFV-WTD), causing malignant catarrhal fever in deer.

In the recent investigation of malignant catarrhal fever in a red brocket deer (Mazama americana) from a Texas zoo, the viral DNA from the herpesvirus termed MCFV-WTD, which causes disease in white-tailed deer (Odocoileus virginianus), was detected. The epidemiology information revealed that the red brocket deer had been associated with a herd of pygmy goats (Capra hircus) at the zoo. MCFV-WTD DNA was also detected in one of these 12 goats that were malignant catarrhal fever viral antibody positive. The amplified herpesviral sequences from the affected deer and the MCFV-WTD-positive goat were identical, and matched the sequence in GenBank. Three of 123 DNA samples from various breeds of goats from different geographic locations in the United States were positive for MCFV-WTD DNA. The study shows that MCFV-WTD is capable of causing malignant catarrhal fever in other species of deer besides white-tailed deer and suggests that goats are a potential reservoir for the virus.

Detection of Mycoplasma agassizii in the Texas tortoise (Gopherus berlandieri).

Mycoplasma agassizii causes upper respiratory tract disease (URTD) in Texas tortoises (Gopherus berlandieri). To determine exposure to and shedding of M. agassizii, we collected blood samples and nasal swabs from 40 free-ranging Texas tortoises on public and private lands in Texas, USA, from May to October 2009. We used an enzyme-linked immunosorbent assay (ELISA) to detect M. agassizii-specific antibodies. Eleven (28%) tortoises were antibody positive, three (8%) were suspect, and the remaining 26 (65%) were negative. Nasal lavage samples were collected from 35 of the 40 tortoises for M. agassizii culture and PCR to detect shedding of M. agassizii. Current infection with M. agassizii was confirmed in one tortoise that had mild clinical signs of URTD and was positive by ELISA (antibody titer >512), PCR, and culture. The clinical isolate was confirmed as M. agassizii by restriction fragment length polymorphism and immunobinding.

Radiation therapy of a malignant melanoma in a thick-billed parrot (Rhynchopsitta pachyrhyncha).

A male thick-billed parrot (Rhynchopsitta pachyrhyncha) was diagnosed with a malignant melanoma of the mandibular beak (gnathotheca). Surgical excision was impossible because of the location of the lesion; a combination of radiation therapy and oral antitumor drugs were used to treat the neoplasm. A whole-body computed tomographic scan showed evidence of metastasis in the lungs; the bird, therefore, was considered to have stage IV disease. Throughout the treatment period, the bird showed no clinical evidence of systemic disease. The bird was given 20 treatments of localized radiation therapy of 2.5 Gray (Gy) for a cumulative dose of 50 Gy. The bird was also treated with piroxicam and cimetidine orally from the time of diagnosis, throughout radiation therapy, and until its death. By the completion of radiation therapy, the initial lesion had decreased considerably in size. The bird survived 2.5 months after radiation therapy was completed but died of complications related to metastatic disease. Necropsy results revealed metastases throughout the body, including lesions in the lungs and liver. To our knowledge, this is the first report of localized radiation therapy and oral antitumor drugs being used to treat malignant melanoma in an avian patient. The radiation therapy did produce tumor response in the form of a reduction in size of the visible tumor.

Patent urachus with subsequent joint infection in a free-living Grevy's zebra foal.

A free-living, female Grevy's zebra (Equus grevyi) foal was found lethargic, lame, with swollen joints, pyrexia, and urine dripping from the umbilicus. It died 2 days later despite intensive care. Gross examination revealed patent urachus and suppurative arthritis. Swabs were taken from the joints, the patent urachus, and urine for bacteriology. The dominant isolate was Escherichia coli. The joint infection was probably secondary to septicemia, resulting from the patent urachus. To our knowledge, this is the first report of neonatal patent urachus in a wild equid.

Serologic survey of selected viral, bacterial, and protozoal agents in captive and free-ranging ungulates from central Kenya.

Serologic evidence of exposure to various disease agents in free-ranging and captive ungulates at a private game ranch in Kenya is presented, and seroprevalence values inside a fenced-in area are compared with those found on the adjacent open savanna. Zebras outside the fence had a higher prevalence of equine rhinovirus-1 than zebras inside (Fisher's exact test, P = 0.007); for all other species and all other agents, there was no such difference (P > 0.10). Results highlight possible transmission of these agents from domestic species into wildlife or vice versa at our study site.