IDENTIFICATION AND RETROSPECTIVE EVALUATION OF A FILARIOID NEMATODE SPECIES IN MANAGED GRASSHOPPER SPARROWS (AMMODRAMUS SAVANNARUM).

The grasshopper sparrow (Ammodramus savannarum) is a species of ground-dwelling passerine bird with 12 different subspecies. The Florida subspecies (Ammodramus savannarum floridanus) is classified as federally endangered, with the most common threats including habitat loss, nest predation, and floods. A managed breeding program was established at White Oak Conservation (Yulee, FL) in 2015 with eastern grasshopper sparrows (Ammodramus savannarum pratensis) as a model for breeding Florida grasshopper sparrows as part of an assurance colony. A filarioid parasite species (Aproctella sp.) was characterized by PCR after identification by blood films and postmortem examinations of both subspecies housed at White Oak Conservation. This Aproctella species was distinct from others with available sequence. Records from 157 eastern and Florida grasshopper sparrows were reviewed, and correlations between presence of filariasis and subspecies, sex, body condition score, and presence of systemic isosporosis, squamous metaplasia, coelomitis, airsacculitis, or a combination of conditions were investigated. Twenty-nine (18.5%) birds (13 of 71 Florida grasshopper sparrows; 16 of 86 eastern grasshopper sparrows) were positive for filariasis by blood film review, grossly or by tissue imprint at postmortem examination, or histologically. Filariasis was significantly correlated with systemic isosporosis, coelomitis, and airsacculitis; was not correlated with subspecies, sex, or squamous metaplasia; and had a questionable correlation with body condition score. This report provides evidence that this Aproctella species has potential to contribute to morbidity and mortality in the grasshopper sparrow. This information will be helpful for implementing effective measures against suspected vectors and for the development of best practice strategies for the health management of the species in breeding programs.

FATAL SYSTEMIC FUNGAL INFECTION IN EASTERN BONGO ANTELOPE (TRAGELAPHUS EURYCERUS ISAACI): SIX CASES.

Over a span of 6 yr, six adult eastern bongo antelope (Tragelaphus eurycerus isaaci) from a single institution died due to systemic mycotic infections. All animals were of the same genetic lineage and in good body condition at the time of death. Gross findings in all cases included multifocal white-to-tan nodules up to 10 cm in diameter that were most numerous in the heart, lung, and kidney. Histologic examination identified these nodules as foci of granulomatous inflammation containing branching, septate, broad, undulating fungal elements. Identification of the fungal species was pursued using PCR with sequencing, immunohistochemistry, and culture. Multiple fungal species were identified using the various modalities, and commonality of species identification was limited to Cladosporium sp. in four of the cases. The clinical and postmortem findings in these cases were identical and were considered to be the same infectious disease. The Cladosporium sp. was considered a candidate as an emerging fatal infectious agent in this population of bongo antelopes. In all of these cases, death was attributed to conduction abnormalities associated with the cardiac lesions or euthanasia.

CLINICAL FINDINGS, PATHOLOGY, BIOSECURITY, AND SEROSURVEILLANCE OF COXIELLOSIS IN WHITE RHINOCEROSES (CERATOTHERIUM SIMUM) AT A CONSERVATION CENTER: TWO CASES.

A primiparous white rhinoceros (Ceratotherium simum) gave birth to a calf overnight after approximately 16 mo of gestation. The calf was found dead in the morning. Necrosuppurative placentitis with bacterial inclusions suggestive of coxiellosis was diagnosed histologically, and Coxiella burnetii was identified in fetal tissues and placenta by polymerase chain reaction and immunohistochemistry. Another primiparous female from the same herd aborted later that year after approximately 15 mo of gestation, and coxiellosis was similarly diagnosed in fetal tissues and on vaginal shedding. Estimates of exposure time, duration of vaginal shedding, and phase I and phase II antibody dynamics were determined retrospectively and prospectively for the two confirmed cases. Biosecurity measures were put in place to prevent guests, staff, and conspecific exposure to the organism. No other confirmed cases have occurred in the collection 3 yr after the initial cases. Coxiellosis outbreaks could represent an emerging threat to conservation efforts and ex situ white rhinoceros breeding programs.

EVALUATION OF A VISUAL ENZYME-LINKED IMMUNOSORBENT ASSAY (ELISA) FOR PREGNANCY DETECTION IN FOUR UNGULATE SPECIES.

This study evaluated the use of a commercially available, visual enzyme-linked immunosorbent assay (ELISA) for diagnosis of pregnancy in okapi (Okapia johnstoni), gerenuk (Litocranius walleri), eastern giant eland (Tragelaphus derbianus spp. gigas), and dama gazelle (Nanger dama). This assay has been validated for use in domestic cattle, sheep, goats, and water buffalo. Unlike other blood-based pregnancy associated glycoprotein (PAG) detection methods, this assay does not require sophisticated laboratory equipment for detection or interpretation and can therefore be utilized in many settings. Banked serum samples from gerenuk (n = 11), giant eland (n = 4), dama gazelle (n = 33) and okapi (n = 3) were tested, and a pregnant and nonpregnant sample from each individual were included. The ELISA showed 100% sensitivity and specificity in gerenuk and giant eland samples, and 0% sensitivity in dama gazelle and okapi samples. Using this assay, pregnancy was detected by 7-8 wk gestation in gerenuk and 6 wk in giant eland. These results are consistent with previous studies that were able to accurately detect pregnancy in other members of the family Bovidae, but it is possible that PAGs present in okapi and dama gazelle are structurally dissimilar relative to the intended test target, and are therefore unrecognizable using this test. The faint positivity in the dama gazelle assays may be due to cross-reactivity with other proteins in the sample, or due to inconsistent binding with the dama gazelle PAG. This ELISA appears to be an accurate, rapid, and inexpensive method of point-of-care pregnancy diagnosis in gerenuk and giant eland, but not okapi and dama gazelle. Additional studies should be pursued to further characterize the limits of pregnancy detection using this assay in gerenuk and giant eland, and to investigate the validity of this test in other nondomestic ruminant species.

SEROLOGICAL EVIDENCE OF COXIELLA BURNETII INFECTION IN THE WHITE RHINOCEROS (CERATOTHERIUM SIMUM) IN SOUTH AFRICA.

Coxiellosis, or Query (Q) fever, a disease caused by the intracellular bacteria Coxiella burnetii, was recently described in a managed breeding herd of white rhinoceros (Ceratotherium simum) in the southeastern United States. Clinical disease often results in abortion and could represent a conservation challenge for this species. In addition to the reproductive and herd management consequences, coxiellosis is also a zoonotic disease. Infection or clinical disease in any free-ranging rhinoceros species in a national park setting has not been previously described. In this study, evidence of prior infection was measured by immunofluorescent antibody titers in 89 serum samples collected from white rhinoceros within private reserves and a national park in South Africa. Total seropositivity was 48/89 (53.9% [95% CI, 43.6-63.9%]). Animals on private reserves had a seropositivity of 21/51 (41.1% [95% CI, 27.1-55.2%]), and national park rhinoceros had a higher rate of seropositivity at 71.0% [95% CI, 55.9-86.2%] (27/38; P= 0.004). Adults had a higher seropositivity compared with subadults (P= 0.03). There was no difference in seropositivity between sexes (P > 0.05). Results demonstrate that South African white rhinoceros populations are exposed to Coxiella, which could result in underrecognized reproductive consequences. Further studies should investigate potential implications for public health and conservation management of this species.

CHRONIC PANCREATIC DISEASE IN THE LESSER KUDU (TRAGELAPHUS IMBERBIS): A REPORT OF 16 CASES IN THE UNITED STATES.

A review of archival cases at Northwest ZooPath from 1995-2018 identified 16/96 (17%; eight females, eight males) lesser kudus (Tragelaphus imberbis) with chronic pancreatic disease (CPD) from three institutions, all of which originated from an initial founder stock of 12 animals. Ages at time of death or euthanasia ranged from 2 to 132 mo (average age = 69 mo). All cases had varying degrees of pancreatic acinar cell depletion, ductular hyperplasia, and fibrosis. Serum fructosamine, glucose, insulin, and insulin to glucose ratio collected close to time of death from 13 affected animals were not significantly different from controls (n = 19). Of these analytes, receiver operating characteristic analysis identified fructosamine as the best-performing analyte with an area under the curve 0.671 (95% confidence interval = 0.480-0.828; sensitivity = 58.3%, specificity = 84.2%; cutoff point >352 µmol/L) in the diagnosis of CPD. With a 15% prevalence, there was a positive predictive value of 41% and a negative predictive value of 92%, indicating that the probability of false positives is high, but of false negatives is low. An etiologic agent was not identified by histology (n = 16), transmission electron microscopy (n = 1), or enterovirus PCR (n = 2). Serum zinc and hepatic heavy metal analyses were judged to be within normal limits. Chronic pancreatic disease is considered an important cause of morbidity and mortality in the lesser kudu; serum chemistry analysis warrants further investigation in its use for diagnosis. The etiopathogenesis is not understood, but the absence of obvious causes, the occurrence also in very young animals, and the inbred lineage of lesser kudus in the United States suggest a genetic basis for this disease.

EVIDENCE OF EPIZOOTIC HEMORRHAGIC DISEASE VIRUS AND BLUETONGUE VIRUS EXPOSURE IN NONNATIVE RUMINANT SPECIES IN NORTHERN FLORIDA.

Epizootic hemorrhagic disease virus (EHDV) and bluetongue virus (BTV) are vector-borne viruses of ruminants nearly worldwide. They can affect white-tailed deer (WTD; Odocoileus virginianus), the ranching industry, and nonindigenous hoof stock species managed for conservation. One potential risk factor for ranched WTD is commingling with nonindigenous species on high-fenced properties. Nonindigenous species provide novel viewing and hunting opportunities; however, their presence may create disease hazards. Furthermore, animals within conservation properties may be at a risk from commingling exotics and adjacent wild WTD. Currently, knowledge about EHDV and BTV seroprevalence and transmission is limited in nonindigenous populations in the southeastern United States. The authors conducted a serological survey of 10 Bovidae and 5 Cervidae species residing within two properties in northern Florida. The first site was a conservation property breeding threatened nonindigenous species for conservation. The second property was a private high-fenced game preserve managing WTD and nonindigenous species for breeding, sale, and harvest. Blood samples were tested for titers to three EHDV serotypes (1, 2, and 6) and active circulating viral EHDV and BTV. The private ranch had evidence of EHDV or BTV in one of three (33.3%) Bovidae species and four of five (80%) Cervidae species sampled. At the conservation property, evidence of EHDV infection was found in four of seven (57.1%) Bovidae and one of one (100%) Cervidae species sampled. The presence of antibodies in many nonindigenous species sampled might indicate these species are potential viral hosts and may be a risk to ranched WTD and other species within the same property. Nonindigenous species within the private ranch and conservation properties are at risk of contracting EHDV and BTV, and herd managers should reduce vector-host interactions and consider increased biosecurity measures when translocating animals.

RETROSPECTIVE EVALUATION OF HEPATOCELLULAR NEOPLASMS IN NILE LECHWE (KOBUS MEGACEROS) FROM TWO FLORIDA ZOOLOGICAL INSTITUTIONS.

This case series describes hepatocellular neoplasms in 10 Nile lechwe (Kobus megaceros) at two separate zoological institutions in Florida. Histologically, the neoplasms were classified as hepatocellular carcinoma (n = 7), hepatocellular adenoma (n = 2), and hepatobiliary carcinoma (n = 1). Common clinical signs were nonspecific and included thin body condition (n =7), lethargy (n =6), lameness (n =3), and acute recumbency (n =5). Four males and six females were affected, and the mean age at death was 12.7 yr with a range of 4-18 yr. All cases were diagnosed postmortem, and metastasis to various sites, including lung, lymph nodes, and omentum, was found in 40% of cases (n = 4). A single case of hepatocellular carcinoma in a Nile lechwe was described in 2007; however, this is the first reported series of neoplasms in Reduncinae. The pathogenesis behind the development of hepatocellular neoplasms in Nile lechwe has not yet been identified.

PHARMACOKINETICS OF A SINGLE ORAL DOSE OF FLUNIXIN MEGLUMINE IN THE WHITE RHINOCEROS (CERATOTHERIUM SIMUM).

Flunixin meglumine, a nonsteroidal anti-inflammatory medication, has been used in rhinoceros species at doses extrapolated from domestic animals. There is increasing evidence to suggest significant variations exist in metabolism of drugs in exotic species. Due to the differences in drug metabolism, dose extrapolation from domestic animals may not be appropriate for exotic species. The objective of this study was to investigate the pharmacokinetics of flunixin meglumine in five white rhinoceroses (Ceratotherium simum) administered a single (1 mg/kg) oral dose of a commercial equine flunixin meglumine paste. Concentrations of flunixin and its metabolite 5-OH flunixin were analyzed, and pharmacokinetic parameters were estimated for each animal. Mean observed plasma concentrations peaked at 1,207 ± 601 ng/ml and occurred at 3 ± 1 hr. The geometric mean of the apparent elimination half-life after oral administration was 8.3 ± 1.2 hr. This data suggests that flunixin meglumine appears to be slowly metabolized or slowly absorbed in this species. No adverse clinical effects were observed during the study period. A single dose of 1 mg/kg appears safe for use in the white rhinoceros. Multidose studies are needed to determine if plasma accumulation of flunixin meglumine occurs and to evaluate safety.

SURGICAL MANAGEMENT OF APPENDICULAR LONG-BONE FRACTURES IN FREE-RANGING FLORIDA PANTHERS ( PUMA CONCOLOR CORYI): SIX CASES (2000-2014).

The clinical outcomes of six free-ranging Florida panthers ( Puma concolor coryi) that underwent surgical stabilization of appendicular long-bone fractures (three femoral fractures, one tibial and one tibial and fibular fracture and two radial and ulnar fractures) were evaluated. These panthers presented to the University of Florida from 2000-2014. Estimated age of the panthers ranged from 0.5 to 4.5 yr, and weights ranged from 22 to 65 kg. Causes of injuries were vehicular collision ( n = 4) and capture related ( n = 2). All panthers underwent open reduction and fracture stabilization. Fixation failure necessitated three subsequent surgeries in one panther. Five panthers survived the immediate postoperative period, and all of these panthers' fractures obtained radiographic union (range, 8-36 [mean, 22] wk). The five surviving panthers underwent convalescence for 7-14 mo at White Oak Conservation Center before being released back into the wild; however, one panther was killed when hit by a car 3 days after release. The remaining four panthers were tracked for up to 106 mo in the wild and successfully integrated back into the native population. Surgical stabilization of appendicular long-bone fractures in free-ranging Florida panthers can be successful, but must take into account the stress that a large, undomesticated felid will place on the stabilized limb during convalescence as well as the difficulties involved in rehabilitating a wild panther in captivity.

CONGESTIVE HEART FAILURE ASSOCIATED WITH PREGNANCY IN OKAPI (OKAPIA JOHNSTONI).

Acute signs associated with cardiovascular disease occurred in three pregnant okapi ( Okapia johnstoni ) during early to midgestation and progressed to congestive heart failure. Congestive heart failure was diagnosed antemortem using echocardiography and plasma cardiac troponin levels. Clinical signs included decreased activity, hyporexia, tachypnea, dyspnea, flared nostrils, and productive coughing with copious amounts of foamy nasal discharge. Parenteral and oral treatment with furosemide, enalapril, and spironolactone controlled clinical signs in the three okapi allowing each to carry out one pregnancy to term. Two okapi carried the first pregnancy to term after showing signs, while one okapi aborted the first calf and gave birth to a healthy calf in a subsequent pregnancy. Subsequent pregnancy in one okapi ended with abortion and associated dystocia and endometritis. Following parturition, clinical signs associated with heart failure resolved in all three individuals; serial echocardiography in two individuals showed improvement in fractional shortening and left atrial size and all three okapi showed markedly decreased pleural effusion and resolution of pulmonary edema. However, subsequent pregnancies in all three okapi induced respiratory distress and recurrence of congestive heart failure; one okapi died from congestive heart failure associated with subsequent pregnancy. This case series describes the clinical presentation and pathologic findings of congestive heart failure during pregnancy in adult okapi.

Pretreatment of Addra gazelle (Nanger dama ruficollis) spermatozoa with cholesterol-loaded cyclodextrins improves cryosurvival.

Preserving genetic diversity of the critically endangered Addra gazelle (Nanger dama ruficollis) could be enhanced through the use of frozen-thawed sperm and artificial insemination. Our aim was to characterize Addra ejaculate traits and to assess the effects of cholesterol-loaded cyclodextrin (CLC) on sperm cryosurvival. Fresh ejaculates were treated with CLC (0, 0.5, 1.5, 3.0, and 6.0 mg/ml) prior to cryopreservation. All males produced spermic ejaculates with >75% sperm motility. The mean ± SEM seminal volume, sperm concentration, percent motility, forward progression, and percent morphologically normal spermatozoa were 3.2 ± 0.3 ml, 1.2 ± 0.3 × 109, 75.82 ± 2.7%, 3.2 ± 0.3 (0-5 scale; 5 = most progressive), and 57.12 ± 3.8%, respectively. More than 92% contained an intact acrosome. There was no effect of time or in vitro incubation on progression or acrosomal integrity on thawed samples (P > 0.05). Spermatozoa pre-treated with 0.5 mg/ml CLC retained higher (P < 0.05) motility post-thaw than aliquots treated with 0, 3.0, or 6.0 mg/ml of CLC. Spermatozoa pre-treated with 0.5, 1.5, or 3.0 mg/ml CLC exhibited greater viability than counterparts (P < 0.05). Sperm kinetics including beat cross frequency (BCF), average path velocity (VAP), curvilinear velocity (VCL), and straight line velocity (VSL) did not differ among samples (P > 0.05). Linearity (LIN) and straightness (STR) were different among samples after thawing. Results demonstrate treatment with CLC (0.5 mg/ml) protects Addra spermatozoa from cryo-damage. Reported advances will facilitate establishment of a frozen repository and support the genetic management of this critically endangered north African desert antelope.

RESULTS OF THE MEGAVEREBRATE ANALGESIA SURVEY: GIRAFFE AND HIPPOPOTAMUS.

Results of an online survey posted on the American Association of Zoo Veterinarians listserv examined the patterns of analgesic medication and pain management modalities used for captive giraffe and hippopotami. Compiled data included signalment, drugs administered, dosing regimens, subjective efficacy scores, ease of administration, and adverse events. Nineteen institutions exhibiting hippopotami ( Hippopotamus amphibious ) and pygmy hippopotami (Choeropsis liberiensis) and 45 exhibiting giraffe ( Giraffa camelopardalis spp.) responded. Phenylbutazone was the most-commonly administered nonsteroidal anti-inflammatory drug (NSAID), followed by flunixin meglumine, but doses varied widely. Eight institutions reported adverse events from NSAID administration. Tramadol was the most-commonly administered opioid followed by butorphanol. Only one adverse event was reported for opioids. Twenty-three of 45 institutions exhibiting giraffe utilized alternative analgesia methods including gabapentin, glucosamine-chondroitin, local anesthetics, and low level laser therapy. Six of 19 institutions exhibiting hippopotami administered omega 3-6 fatty acids, gabapentin, glucosamine-chondroitin, and α-2 adrenergics to provide analgesia. While all reporting zoological institutions administered similar drugs, there was substantial variation and diversity in both dosing regimens and frequencies, indicating the need for both preclinical and clinical studies supporting dosing regimens.

RESULTS OF THE MEGAVERTEBRATE ANALGESIA SURVEY: ELEPHANTS AND RHINO.

An online survey utilizing Survey Monkey linked through the American Association of Zoo Veterinarians listserve examined current practices in megavertebrate analgesia. Data collected included drugs administered, dosing regimens, ease of administration, efficacy, and adverse events. Fifty-nine facilities (38 housing elephants, 33 housing rhinoceroses) responded. All facilities administered nonsteroidal anti-inflammatory drugs (NSAIDs), with phenylbutazone (0.25-10 mg/kg) and flunixin meglumine (0.2-4 mg/kg) being most common. Efficacy was reported as "good" to "excellent" for these medications. Opioids were administered to elephants (11 of 38) and rhinoceroses (7 of 33), with tramadol (0.5-3.0 mg/kg) and butorphanol (0.05-1.0 mg/kg) being most common. Tramadol efficacy scores were highly variable in both elephants and rhinoceroses. While drug choices were similar among institutions, substantial variability in dosing regimens and reported efficacy between and within facilities indicates the need for pharmacokinetic studies and standardized methods of analyzing response to treatment to establish dosing regimens and clinical trials to establish efficacy and safety.

Critical care for a hypothermic and hypoglycemic white rhinoceros (Ceratotherium simum simum) calf.

A 3-day-old, 57.3-kg, male white rhinoceros (Ceratotherium simum simum) neonate presented laterally recumbent with comatose mentation, cold extremities, and severe hypothermia, hypoxemia, and hypoglycemia. Critical care support was initiated with aggressive fluid and warming support as well as dextrose and oxygen supplementation. After initial stabilization, additional complications arose in subsequent weeks including intermittent diarrhea, dry skin with loss of epidermal layers, urticaria on the head, and superficial wounds on the face, ears, feet, and penis. There is a lack of critical care information for rhinoceros calves. However, due to similarities to other Perissodactylids, some general guidelines for critical equid neonatal care were implemented. The calf was hand-raised until weaning and is now a subadult white rhinoceros with no abnormalities.

Comparison of noninvasive blood pressure measurement techniques via the coccygeal artery in anesthetized cheetahs (Acinonyx jubatus).

Two indirect blood pressure measurement techniques, Doppler (DOP) sphygmomanometry and oscillometry, applied at the ventral coccygeal artery were compared with simultaneous direct blood pressure measurements at the dorsal pedal artery in 10 anesthetized, captive cheetahs (Acinonyx jubatus). The DOP method was moderately accurate, with relatively little bias (mean difference 3.8 mmHg) and 88.6% of the DOP systolic arterial pressure measurements being within 10 mmHg of the direct systolic arterial measurement. With the oscillometric (OM) method, 89.2% of the mean arterial pressure measurements were within 10 mmHg of the direct measurement and had the least bias (mean difference 2.3 mmHg), 80.7% of the systolic measurements were within 10 mmHg of the direct measurement and had the second least bias (mean difference 2.3 mmHg), and 59% of the diastolic measurements were within 10 mmHg of the direct measurement and had significant bias (mean difference 7.3 mmHg). However, DOP showed relatively poor precision (SD 11.2 mmHg) compared with OM systolic (SD 8.0 mmHg), diastolic (SD 8.6 mmHg), and mean (SD 5.7 mmHg). Both techniques showed a linear relationship with the direct technique measurements over a wide range of blood pressures. The DOP method tended to underestimate systolic measurements below 160 mmHg and overestimate systolic measurements above 160 mmHg. The OM method tended to underestimate mean pressures below 160 mm Hg, overestimate mean pressures above 160 mmHg, underestimate systolic pressures below 170 mmHg, overestimate systolic pressures above 170 mmHg, and underestimate diastolic pressures throughout the measured blood pressure range. Indirect blood pressure measurement using the ventral coccygeal artery, particularly when using an OM device for mean and systolic arterial pressure, may be useful in the clinical assessment of cheetahs when monitoring trends over time, but caution should be taken when interpreting individual values.

Surgical resolution of an avulsion fracture of the peroneus tertius origin in a giraffe (Giraffa camelopardalis reticulata).

A 4-mo-old, 185-kg male giraffe (Giraffa camelopardalis reticulata) was presented due to stifle effusion and lameness of 3-wk duration. Radiographs revealed a fracture of the extensor fossa of the lateral trochlear ridge of the femur at the origin of the peroneus tertius. Under anesthesia, dysfunction of the reciprocal apparatus was documented by flexing the stifle while the tarsus remained extended. An avulsion fracture of the origin of the peroneus tertius and extensor digitorum longus muscle was diagnosed. An exploratory arthroscopy of the femorotibial joint was followed by arthrotomy to excise the large bone fragment from its soft tissue attachments. Because of the fractious temperament of the animal, postoperative care was restricted to stall rest for 3 mo, and no postoperative complications arose. Only a mild residual lameness remained by 6 mo after surgery.

Serum chemistry comparisons between captive and free-ranging giraffes (Giraffa camelopardalis).

Serum chemistry analyses were compared between captive and free-ranging giraffes (Giraffa camelopardalis) in an attempt to better understand some of the medical issues seen with captive giraffes. Illnesses, including peracute mortality, energy malnutrition, pancreatic disease, urolithiasis, hoof disease, and severe intestinal parasitism, may be related to zoo nutrition and management issues. Serum samples were collected from 20 captive giraffes at 10 United States institutions. Thirteen of the captive animal samples were collected from animals trained for blood collection; seven were banked samples obtained from a previous serum collection. These samples were compared with serum samples collected from 24 free-ranging giraffes in South Africa. Differences between captive and free-ranging giraffes, males and females, and adults and subadults were analyzed by using a 2 x 2 x 2 factorial and Fisher's least significant difference for mean separation; when necessary variables were ranked and analyzed via analysis of variance. Potassium and bilirubin concentrations and alanine aminotransferase (ALT) activities were different between captive and free-ranging giraffes, but all fell within normal bovid reference ranges. The average glucose concentration was significantly elevated in free-ranging giraffes (161 mg/dl) compared with captive giraffes (113 mg/dl). All giraffes in this study had glucose concentrations higher than bovine (42-75 mg/ dl) and caprine (48-76 mg/dl) reference ranges. Differences were also seen in lipase, chloride, and magnesium though these findings are likely not clinically significant. There were no differences detected between sexes. Adults had higher concentrations of potassium, total protein, globulins, and chloride and higher gamma glutamyltransferase activities, whereas subadults had higher concentrations of phosphorus. Within the captive group, nonimmobilized animals had higher concentrations of total protein and globulins. Captive giraffe diets need further investigation to determine if the differences seen in this study, especially glucose and bilirubin concentrations and ALT activities, may result in some health problems often seen in captive giraffes.

Full Viral Genome Sequencing and Phylogenomic Analysis of Feline Herpesvirus Type 1 (FHV-1) in Cheetahs (Acinonyx jubatus).

Feline herpesvirus type 1 (FHV-1) is endemic in captive cheetahs and sporadically causes devastating disease. Modified live vaccines (MLV), intended for use in domestic cats, are used in some captive cheetah populations and have been anecdotally linked to disease in certain subpopulations. Ten FHV-1 isolates from ten captive cheetahs and one isolate from an MLV used to inoculate four of the host animals were analyzed. Viral DNA was extracted for full-genome sequencing by Illumina MiSeq with viral genomes then used for phylogenomic and recombinational analyses. The FHV-1 shed by vaccinated cheetahs were almost identical to the MLV, with few variants among viral genomes. Eight cheetah FHV-1 isolates and the MLV were grouped in a clade along with FHV-1 isolates from domestic cats in the USA. The remaining two cheetah FHV-1 isolates (unknown host vaccine status) were not associated with a clade. The likely ancestral origin of these two isolates involves recombination events between Australian domestic cat and cheetah FHV-1 isolates. Collectively, these data suggest that the MLV is capable of causing clinical disease and viral shedding in some cheetahs and represents evidence of interspecies transmission of virus between domestic and wild cats.

Characterization of an outbreak of astroviral diarrhea in a group of cheetahs (Acinonyx jubatus).

A Mamastrovirus was identified in an outbreak of diarrhea in cheetahs (Acinonyx jubatus). Five young adult and two adult cheetahs presented with lethargy, anorexia, watery diarrhea and regurgitation over an 11-day period. Fecal samples were submitted for electron microscopy and culture. Electron microscopy results revealed particles morphologically consistent with an astrovirus, and no other viral pathogens or significant bacterial pathogens were identified. The astrovirus was confirmed and sequenced using consensus astroviral PCR, resulting in a 367 base pair partial RNA-dependent-RNA polymerase (RdRp) product and a 628 base pair partial capsid product. Bayesian and maximum likelihood phylogenetic analyses were performed on both the RdRp and the capsid protein segments. All animals were monitored and treated with bismuth subsalicylate tablets (524mg PO BID for 5 days), and recovered without additional intervention. This is the first report we are aware of documenting an astrovirus outbreak in cheetah.