Cellular immune function in four rhinoceros species was evaluated by way of in vitro lymphocyte proliferation responses to mitogenic and antigenic stimuli to establish normative data on white blood cell activity for each species and to identify species-specific differences that might help explain the predisposition of black rhinoceroses (Diceros bicornis) to disease. A cross section of the U.S. rhinoceros population encompassing all four captive species was sampled, including the Sumatran rhinoceros (Dicerorhinus sumatrensis) (n = 3); Indian rhinoceros (Rhinoceros unicornis) (n = 4); African black rhinoceros (n = 16); and African white rhinoceros (Ceratotherium simum) (n = 10). Of the four species evaluated, African black rhinoceroses exhibited the weakest (P < 0.05) lymphocyte proliferative responses to the mitogens: pokeweed (0.1 μg/ml), phytohemagglutinin (0.3 μg/ml), and concanavalin A (5.0 μg/ml). Total cell density at the end of culture was only 70% of that achieved with lymphocytes isolated from African white rhinoceroses, Indian rhinoceroses, and Sumatran rhinoceroses. However, lymphocyte response to bacterial endotoxin lipopolysaccharide was similar (P > 0.05) across species. Antigenic stimulation produced much weaker responses than mitogenic stimulation. No differences (P > 0.05) were observed among rhinoceros species in response to 1 and 10 μg/ml of Leptospira icterohemorrhagiae or Leptospira gryppotyphosa. Lymphocytes from African white rhinoceroses proliferated weakly in the presence of Aspergillus fumigatus filtrate, whereas lymphocytes from the southern black rhinoceros subspecies appeared slightly suppressed in the presence of increasing doses (0.1, 1, and 10 μg/ml) of Aspergillus filtrate. This comparative data set characterizing lymphocyte proliferation in the rhinoceros reveals several differences in immune cell responses among rhinoceros species and provides some evidence that lymphocytes of captive African black rhinoceroses are less vigorous than those of the other rhinoceros species.

The common piping guan (Pipile cumanensis cumanensis) is serving as a model for assisted reproductive techniques (ARTs) in Cracidae (Aves: Galliformes). Manual collection methods used in domestic birds and other exotics were modified to optimize quality and quantity of fresh semen samples. In addition, a method to subjectively assess semen quality on the basis of its color, clarity, and consistency was developed to serve as a tool when laboratory equipment is not available. In a species where only two eggs per clutch is typical, predictability of the date of the first egg is critical to ensure adequate inseminations before oviposition. An increase in both the distance between the female's pubic bones and degree of abdominal swelling indicated optimal time to initiate artificial insemination (AI) and allowed the number of semen samples used to fertilize the first egg in the clutch to be minimized while ensuring its fertility. In three separate AI trials, a total of four fertile eggs were produced, a first for cracids. Two of the four fertile eggs hatched successfully, and the chicks' parentages were confirmed using DNA fingerprinting. This study represents the first published account of the use of ARTs in Cracidae and forms the basis for future developments (e.g., AI of frozen– thawed semen) aimed at optimizing genetic management of cracids in ex situ breeding programs.

Seventeen Gulf of Mexico sturgeons (Acipenser oxyrinchus desotoi) underwent endoscopic sex determination, gonadal biopsy, and various reproductive surgeries as part of a conservation development plan. The fish were anesthetized with tricaine methanesulfonate (MS-222) buffered with sodium bicarbonate and maintained on a recirculating water anesthesia circuit. A 6-mm Ternamian EndoTip Cannula, placed through the ventral midline, midway between pectoral and pelvic fins, permitted the introduction of a 5-mm telescope. Swim bladder aspiration and CO₂ insufflation of the coelomic cavity provided excellent observation. Second and third cannulae were placed under direct visual control, lateral and cranial or caudal to the telescope cannula. Sex determination was successfully performed in all fish; however, five of 17 sturgeons (29%) required endoscopic gonadal biopsy to confirm sex. Bilateral ovariectomy or orchidectomy was successfully performed in three males and four females. Unilateral ovariectomy and bilateral ligation of the müllerian ducts using an extracorporeal suturing technique was accomplished in an additional three females. No apparent morbidity was associated with the anesthesia or endoscopic surgery in any fish. The ability to safely perform minimally invasive reproductive surgery in fish may have important management and conservation benefits.

Stingrays are prominent marine animals; however, there are few published reference values for their blood chemistry and hematology. Twenty-eight southern stingrays (Dasyatis americana) were caught using the bottom trawl nets of fishery-independent boats operated by the South Carolina Department of Natural Resources during June and July 2002 from Winyah Bay, South Carolina, to St. Augustine, Florida. Median values of blood and plasma obtained from live animals promptly after capture are as follows: packed cell volume = 0.22 L/L (22%), total solids (TS) = 56.5 g/L (5.65 g/dl), total protein (TP) = 26 g/L (2.6 g/dl), sodium = 315 mmol/L, potassium = 4.95 mmol/L, chloride = 342 mmol/L, calcium = 4.12 mmol/L (16.5 mg/dl), phosphorus = 1.5 mmol/L (4.7 mg/dl), urea nitrogen = 444 mmol/L (1,243 mg/dl), glucose = 1.69 mmol/L (30 mg/dl), aspartate aminotransferase = 14.5 U/L, creatine phosphokinase = 80.5 U/L, osmolality = 1065 mOsm/kg, and lactate = 3.1 mmol/L. Bicarbonate was less than the low end of the instrument range (5 mmol/L) in all but three samples. Anion gap was negative in all samples. Albumin was less than the low end of the instrument range (1 g/dl) in all except one sample. Osmolality was significantly higher in the rays caught in the southern region. TS and TP values were linearly related to each other, and the equation for the fitted line is TS = (11.61 × TP) 25.4 (in g/L) [or TS = (1.161 × TP) 2.54 (in g/dl)]. The reference ranges reported in this study can be used to aid in the management of aquarium stingrays and to create a baseline for health monitoring of the wild Dasyatis spp.

The pharmacokinetics of oxytetracycline in 2-yr-old loggerhead sea turtles (Caretta caretta) after single i.v. and i.m. injections were studied for biologic marking and therapeutic applications. Twenty juvenile turtles were divided into two treatment groups. Ten animals received 25 mg/kg of oxytetracycline i.v. and 10 received the same dosage i.m. Plasma oxytetracycline concentrations were analyzed by reverse-phase high-performance liquid chromatography. Data from the i.v. route best fit a three-compartment model, whereas noncompartmental analysis was used to compare data from both the i.v. and i.m routes. For the i.v. route, means for maximum plasma concentration, terminal phase half-life, systemic clearance, and apparent volume of distribution at steady state were 6.6 μg/ml, 66.1 hr, 290.7 ml/hr/kg, and 18.4 L, respectively. For the i.m. route, means for systemic availability, maximum plasma concentration, and elimination half-life were 91.8%, 1.6 μg/ml, and 61.9 hr, respectively. The remarkably high apparent volume of distribution may possibly be associated with a deep compartment of drug disposition such as bone deposition associated with the large skeletal mass of turtles and the fact that these were well-nourished, growing juveniles. Although maximum plasma concentration by i.m. administration was lower than for the i.v. route, the long elimination time indicates that an infrequent dosing interval may be effective for sensitive bacteria.

Using a crossover study design, the pharmacokinetics of carfentanil and naltrexone after i.v., i.m., and s.c. administration were determined in eight domestic goats (Capra hircus). Serial blood samples were taken up to 120 hr after carfentanil administration, and the plasma drug concentrations were determined using liquid chromatography and mass spectroscopy. All goats were immobilized with 40 μg/kg carfentanil i.m., although the resulting neurologic effects varied considerably. Plasma profiles showed rapid carfentanil absorption and a simple biphasic decline for 12– 48 hr. Naltrexone given at 100 mg naltrexone/mg carfentanil 30 min after carfentanil administration produced rapid reversal of immobilization after all routes of administration. Variable fluctuations in the naltrexone plasma concentrations during the first 2.5–3.5 hr were observed, followed by a more consistent biphasic decline. The time to standing was significantly shorter after i.v. compared with s.c. naltrexone, although the time difference (1 min) had little clinical relevance. No statistically significant differences between the naltrexone pharmacokinetic parameters measured for the three routes of naltrexone administration were identified, although the recoveries after i.m. administration were, subjectively, the smoothest. The carfentanil half-life did not differ significantly in the goats given naltrexone by different routes. Although it is currently recommended that the naltrexone dose be divided into s.c. and i.v. portions, this practice does not appear to offer any benefit.

The efficacies and ease of administration of four oral preanesthetic sedation protocols were compared in 18 adult, male rhesus macaques (Macaca mulatta) to achieve heavy sedation and alleviate anxiety, agitation, and potential trauma associated with remote anesthesia induction. The macaques, with average age and weight of 10 yr and 12.5 kg, respectively, were randomly assigned to one of four groups. Group 1 was given 10 mg/kg tiletamine– zolazepam and 0.05 mg/kg medetomidine p.o., group 2 was given 1 mg/kg midazolam and 20 mg/kg ketamine p.o., group 3 was given 20 mg/kg ketamine and 0.05 mg/kg medetomidine p.o., and group 4 was given 3 mg/kg midazolam p.o. All protocols produced effects ranging from mild sedation to no response to noxious stimuli, depending on the success of administration. The mean interval to peak effect was 27–43 min in all groups. Ketamine and medetomidine provided significantly better sedation than midazolam alone; there were no other statistically significant differences among the four protocols. Oral tiletamine–zolazepam and medetomidine provided smooth, mild to moderate sedation with few side effects. The midazolam and ketamine combination resulted in severe ataxia. Orally administered ketamine and medetomidine provided smooth, easily reversible, heavy sedation leading to no response to noxious stimuli. Midazolam alone provided only mild sedation. No statistically significant differences in palatability of the four protocols were identified. Orally administered ketamine and medetomidine (group 3) provided the most consistently heavy sedation. A compounding pharmacy may be able to increase the palatability and level of acceptance of these combinations. Alternatively, oral midazolam syrup is well accepted by some animals and provides a mild sedative and calming effect, which may decrease stress associated with the induction of anesthesia via darting, pole syringes, etc.

In recent years, several species of ehrlichiae have been recognized as tick-borne disease agents of veterinary and medical importance. Clinically normal free-ranging or previously free-ranging lemurs, including 46 ring-tailed lemurs (Lemur catta), six blue-eyed black lemurs (Eulemur macaco flavifrons), and four black and white ruffed lemurs (Varecia variegata variegata) from St. Catherines Island, Georgia, were tested for evidence of exposure to tick-borne ehrlichiae. All 52 adult lemurs were serologically tested for exposure to Ehrlichia chaffeensis and Anaplasma phagocytophilum. Polymerase chain reaction (PCR) assays for E. chaffeensis, A. phagocytophilum, Ehrlichia ewingii, and Ehrlichia canis were conducted on blood samples from all 56 lemurs. Blood from all lemurs was inoculated into DH82 cell cultures for E. chaffeensis isolation. Of the adult lemurs, 20 (38.5%) and 16 (30.8%) had antibodies reactive (≥1:128) for E. chaffeensis and A. phagocytophilum, respectively. Two ring-tailed lemurs were PCR and culture positive for E. chaffeensis. Molecular characterization of the two E. chaffeensis isolates showed that both contained 5-repeat variants of the variable-length PCR target (VLPT) antigen gene and 3-repeat variants of the 120-kDa antigen gene. Sequencing of the VLPT genes revealed a novel amino acid repeat unit (type-9). One lemur infected with E. chaffeensis was slightly hypoproteinemic and had moderately elevated serum alanine aminotransferase (ALT) levels. These lemurs from St. Catherines Island have been exposed to or infected with tick-borne ehrlichiae, or both, but showed no clinical disease.

Standard limb, six lead (I, II, III, aVR, aVL, and aVF) electrocardiograms (ECGs) were recorded in 10 awake mature rock partridges (Alectoris graeca) and 10 chukar partridges (Alectoris chukar). Durations and amplitudes of P and T waves and QRS complexes, durations of P-Q and Q-T intervals, and mean heart rates were calculated from the lead II ECGs. All observable P and T waves were negative in aVR and aVL, whereas they were positive in all remaining leads. The most frequent forms of QRS complex were r-s (r-S) and q-r (q-R). A Q wave was observed in all aVR and aVL leads in both species. Chukar partridges had significantly higher amplitudes of P and T waves and QRS complexes than rock partridges. Mean heart rates were 310 ± 15 beats/min and 317 ± 19 beats/min for chukar partridges and rock partridges, respectively. Mean electrical axes, calculated from leads II and III, were −99 ± 6.3° and −95 ± 1.7° for chukar partridges and rock partridges, respectively. Clear ECGs were easily obtainable without anesthesia or sedation.

A total of 20 (14 females, six males) captive striped skunks (Mephitis mephitis) with miniature temperature dataloggers implanted free in the abdominal cavity were examined for reproductive performance and pathology because of implants. Eleven of 12 female skunks reproduced successfully 45.9 ± 3.7 days after surgery to remove implanted dataloggers. The pregnancy rate of 91.7% (11/12) was much higher than other captive studies and was comparable with that of wild skunks. Eight striped skunks (six males, two females) that were euthanatized and necropsied after having implants in the abdominal cavity for 5 mo showed no apparent pathology associated with the implant. Implantation of the intraperitoneal devices did not lead to complications in abdominal tissues. Neither implant nor surgery affected reproduction. We conclude that implanted dataloggers can safely be used in physiologic studies of striped skunks or possibly other small carnivores in captive or field studies.

Two juvenile, male, captive-born lowland gorillas (Gorilla gorilla gorilla) with the same father but different mothers developed bilateral cataracts. The cataracts were surgically removed within 6 yr and 3 mo, respectively, of diagnosis, and foldable intraocular lenses were implanted. Although vision was not restored in one eye with a mature, 6-yr-old cataract in gorilla A, surgical intervention on the other eye was performed before a complete cataract developed, and vision was fully restored. Gorilla B was treated at the age of 17 mo, and normal visual development proceeded in both eyes. This animal developed bilateral after-cataract and therefore needed a second intervention in both eyes using Nd:YAG laser treatment. The genetic component of juvenile cataracts should be considered in breeding management programs.

Three juvenile male Micronesian kingfishers (Halcyon cinnamomina cinnamomina) housed in the same enclosure presented with rapid weight gain and coelomic distension. Physical examination and radiography revealed marked enlargement of the ventriculus and a single, large foreign body within the ventriculus in each individual. Surgical removal by ventriculotomy was attempted in one individual, which died during the procedure. A second individual was treated with natural peanut butter 0.5 ml p.o. b.i.d. for 14 days and recovered, casting the foreign material. The third bird was similarly treated without success and subsequently died during attempts at endoscopic removal of the foreign body. In all three birds, the foreign bodies proved to be phytobezoars. The birds had been observed stripping leaf fragments from live corn plants (Dracaena fragrans) used in the enclosure. Plant fibers from the phytobezoars were compared with D. fragrans leaves and were considered identical. Medical treatment of phytobezoars with peanut butter or similar oil-containing substances in birds should be considered as an alternative to surgical extraction.

Three captive female nilgais (Boselaphus tragocamelus) either showed late-term abortion or their newborn calves died within 2 days of birth. Toxoplasma gondii DNA was demonstrated in the brain and liver of each fetus and in one of the two neonates by single-stage polymerase chain reaction (PCR) with TGR1E and by seminested PCR with B1 gene. Retrospectively, antibodies titers ≥640 to T. gondii by indirect fluorescence were found in the sera of all three female and one male nilgais. No other cause of abortion was detected. Fatal toxoplasmosis was also diagnosed in one captive, adult female saiga antelope (Saiga tatarica), which died suddenly. Toxoplasma gondii was detected in the liver, lung, spleen, kidney, and intestine. An unusual finding was the presence of numerous tissue cysts in the liver of this animal. Toxoplasmosis was confirmed by PCR with TGR1E and immunohistochemically. Toxoplasmic hepatitis and pneumonia were considered to be the primary causes of death.

Seven captive eastern box turtles (Terrapene carolina carolina) from a large collection of North American chelonians in North Carolina became acutely ill in the fall of 2002. Five of the turtles died. Clinical signs included cutaneous abscessation, oral ulceration or abscessation (or both), respiratory distress, anorexia, and lethargy. The predominant postmortem lesion was fibrinoid vasculitis of various organs, including skin, mucous membranes, lungs, and liver. No inclusion bodies were detected by histopathology or electron microscopy of formalin-fixed tissue. An iridovirus was isolated from tissues obtained postmortem from two of the box turtles that died. The virus was characterized by electron microscopy, polymerase chain reaction, and sequence analysis of a portion of the major capsid protein as a member of the genus Ranavirus.

Four aardvarks (Orycteropus afer) were anesthetized over a 6-yr period. They were sedated using detomidine (0.13 ± 0.025 mg/kg i.m.; 0.12–0.14 mg/kg) and anesthetized with detomidine (0.12 ± 0.025 mg/kg i.m.; 0.09– 0.18 mg/kg) followed by ketamine (6.3 ± 1.68 mg/kg i.m.; 4.3–8.2 mg/kg). Effects of anesthesia were reversed by atipamezole (0.065 ± 0.013 mg/kg i.m.; 0.05–0.09 mg/kg). The detomidine–ketamine combination produced smooth anesthesia, excellent muscle relaxation, and was suitable for routine diagnostic and therapeutic interventions (blood collection, radiologic examinations, minor surgery).

An 8-yr-old male meerkat (Suricata suricatta) presented with sudden lethargy. A globular-shaped heart associated with pleural effusion on chest thoracic radiographs was consistent with congestive heart failure, and echocardiography was performed under general anesthesia. It revealed an Ebstein anomaly, with mild pericardial effusion and marked right heart enlargement. The animal was treated with imidapril chlorydrate. After 4 mo of treatment, thoracic radiographs still showed right-sided cardiomegaly; however, the animal appeared clinically normal.

A 2.5-yr-old, intact male Japanese macaque (Macaca fuscata) was observed to have a thickened ileum during exploratory laparotomy. Lawsonia intracellularis–associated proliferative enteritis was diagnosed using histopathology (Warthin–Starry stain), immunohistochemistry, and polymerase chain reaction analysis of the ileal biopsy. The animal developed transient diarrhea and severe hypoproteinemia 16 days after surgery but recovered with intensive treatment using azithromycin. Given the fact that very specific tests are required for identifying this organism, L. intracellularis may be underdiagnosed in nonhuman primates.

A mass was removed from the left flank of a 10-yr-old male king cobra (Ophiophagus hannah), and histologic examination revealed granulomatous dermatitis with intralesional gram-positive cocci and filamentous bacteria. Fourteen months later, a histologically similar subcutaneous mass was removed from a different site. One year later, a large subcutaneous mass at the first surgical site was removed, and histopathologic examination revealed multiloculated granulomas with intralesional gram-positive cocci. An organism was cultured and identified by 16S ribosomal RNA gene sequencing as Dermatophilus chelonae. After a course of antibiotic therapy, no further lesions were seen for 5 mo.

Two dead, captive green anacondas (Eunectes murinus), including one male and one female, submitted for necropsy were in poor body condition, having multiple, scattered, dark red foci on the scales and mottled lungs. Both snakes had severe mycotic dermatitis. In addition, the male snake had mycotic stomatitis, and the female snake had mycotic pneumonia. Trichophyton sp., Verticillium sp., and Alternaria sp. were isolated from the dermal lesions. The pulmonary lesions were morphologically consistent with Aspergillus sp. Bacterial organisms isolated from skin and internal organs included Chryseobacterium meningosepticum, Stenotrophomonas maltophilia, Aeromonas hydrophila, and Providencia rettgeri. Mycotic diseases can be devastating to reptiles, and suboptimal husbandry and captivity were likely the predisposing factors that led to opportunistic invasion in these snakes.

Salmonellosis is an important reptile-associated zoonotic infection in the United States. Cloacal swabs were collected from reptiles admitted to the Wildlife Center of Virginia, Waynesboro, Virginia, cultured for Salmonella using Hektoen and xylose lysine deoxycholate agars, and inoculated in selenite broth. All three were incubated at 37°C for 18–24 hr. Seventy-five animals were included in the study, representing eight species, 34 eastern box turtles (Terrapene carolina carolina), 14 eastern painted turtles (Chrysemys picta picta), 14 snapping turtles (Chelydra serpentina), 6 black rat snakes (Elaphe obsoleta obsoleta), 2 redbelly turtles (Pseudemys rubriventris), 2 yellowbelly sliders (Trachemys scripta scripta), 2 eastern garter snakes (Thamnophis sirtalis sirtalis), and 1 eastern river cooter (Pseudemys concinna concinna). All cultures were negative for Salmonella spp., which is in contrast to the high prevalence of Salmonella cloacal shedding reported in captive reptiles but similar to previous reports in free-living North American reptiles. We recommend, nonetheless, practicing proper hygiene methods when handling and housing all reptiles.

A 4.5-yr-old male dromedary camel (Camelus dromedarius) was evaluated for a mass on the right side of the face. A complete blood count and blood chemistry revealed anemia and hypoproteinemia. Radiographs did not reveal bony involvement. The mass was resected and Pythium insidiosum was cultured. The camel was treated with an experimental immunotherapeutic vaccine and with sodium iodide and ceftiofur. The camel began to lose weight postoperatively and died 6 mo later. At necropsy, the camel was found to have gastritis of the third compartment of the stomach with intralesional hyphae of this oomycete pathogen.

An elastic tape sling successfully maintained reduction of a craniodorsal right coxofemoral luxation in a woodchuck (Marmota monax). No complications were encountered during the 14 days that the sling was in place. The animal remained normal through the following 8 mo.

A 3-yr-old intact male dingo (Canis familiaris dingo) presented with a 3-mo history of diarrhea. The diarrhea did not resolve with antibiotics or intestinal protectants. Fecal examination for parasites, fecal cultures, physical examination, and radiographs were unremarkable. Enteroscopic duodenal biopsies showed dilated lacteals without inflammation. Results of serum folate, cobalamin, and trypsin-like immunoreactivity were normal. Low serum total protein and albumin combined with increased fecal levels of alpha-1 protease inhibitor suggested the diagnosis of lymphangiectasia. Full-thickness intestinal biopsies of the duodenum, jejunum, and ileum revealed dilated mucosal and submucosal lacteals without associated inflammation, confirming the diagnosis of primary lymphangiectasia. Currently, the dingo is being maintained with nutritional management.

A 1.5-kg, 6-mo-old male patas monkey (Erythrocebus patas) was cyanotic and panting. Evaluation of the heart by electrocardiography, thoracic radiography, two-dimensional echocardiography, and Doppler color-flow echocardiography revealed a ventricular septal defect (VSD) with right-to-left shunting as well as tricuspid valve dysplasia with dilated annulus of the tricuspid ring, dilated right atrium, dilated right ventricle, and deformity of the tricuspid valve. Because of the severity of the cardiac disease, the patas monkey had complications recovering from anesthesia and died 3 days later. Gross postmortem findings included VSD, tricuspid dysplasia, and cerebral hemorrhage.