Fibropapillomatosis continues to be an important cause of morbidity and mortality in sea turtles, particularly in Chelonia mydas. Turtles with this debilitating herpesvirus disease usually present with multiple, large, and ulcerated cutaneous masses that compromise both locomotion and feeding. There are very few available therapeutic strategies, with surgical excision being the most common. However, this surgical excision is associated with a high rate of local disease recurrence and secondary infections. Electrochemotherapy has been used for the treatment of epithelial neoplasm in several animal species. This technique is based on a combination of chemotherapy, usually with bleomycin or cisplatin, and electroporation. It consists of a series of short, high-voltage electric pulses that lead to increased membrane permeability and more efficient transport of antineoplastic drugs through the cellular membrane. Here, two C. mydas fibropapillomas were treated with a standard electrochemotherapy protocol using intralesional bleomycin sulfate injections followed by the application of electric pulses. Two sessions were performed, with a 33-day interval between sessions. Complete regression of lesions occurred without side effects or complications in each animal. There was no sign of local recurrence, even 1 yr after the end of treatment. Electrochemotherapy may be an effective therapeutic alternative for sea turtles with fibropapillomas.

Members of the genus Iridovirus (invertebrate iridoviruses [IIVs]) of the Iridoviridae family infect a wide range of invertebrates, mainly arthropods, but there have also been a few reports from other taxa. The cricket iridovirus described recently has been shown to infect a wide host range among insect orders and has also been described in several diseased reptiles. This virus together with the type species Chilo iridescent virus form a distinct “group II” in the genus. The aim of this study was to develop a fast and easy real-time polymerase chain reaction [quantitative (q)PCR] for the detection of these group II iridoviruses. In silico and in vitro assays demonstrated that the designed TaqMan primer–probe combination targeting a portion of the major capsid protein is specific for this group of IIVs. A sensitivity assay showed that it is able to detect as little as one copy of viral DNA. Direct comparison of cell culture isolation, nested (n)PCR, and qPCR methods has shown that PCR methods are 10²–10³ more sensitive compared with the isolation method. In testing the three methods on routine diagnostic samples from lizards (n = 22) and crickets (n = 11), the nPCR and qPCR results were consistent with 19 positive lizards and 10 positive crickets, respectively, whereas isolation on cell culture detected only seven and six positives, respectively. QPCR is a fast, sensitive, and specific diagnostic method. Furthermore, it requires fewer handling steps than were previously required. It also allows the quantitation and comparison of the amounts of IIV DNA in samples.

Reference intervals for blood biochemical analytes and packed cell volume are useful to veterinarians and conservation programs for monitoring the health of threatened species. Populations of the critically endangered Chinese three-striped box turtle, Cuora trifasciata (Bell, 1825), are heavily depleted in the wild because of overharvesting; and many individuals are now maintained in captive breeding programs. Reference intervals of biochemical analytes and packed cell volume were established for individuals of this species held in a captive breeding program at Kadoorie Farm and Botanic Garden, Hong Kong. Blood samples were obtained from 86 healthy individuals from a captive population during annual health examinations between 2006 and 2010. Packed cell volume and 17 biochemical analytes were measured, and data were analyzed using both multivariate and univariate statistical analyses. Significant differences in eight analytes between males and females were identified. Ten analytes were found to increase or decrease significantly with the body weight of the turtles. The reported plasma reference intervals, based on 2.5th and 97.5th percentiles, may serve as benchmarks for clinical assessment and conservation of this critically endangered species.

Animals from seven zoological gardens located in Romania, including 18 species of herbivores, 10 species of carnivores, and 13 species of omnivores, were screened for the presence of parasites. Overall, the prevalences of parasites identified in the sampled population were 54.2% (58/107) for herbivores, 54.5% (24/44) for carnivores, and 32.6% (17/52) for omnivores. In herbivores, Eimeria spp., Dicrocelium lanceolatum, and pulmonary and digestive strongyles were detected. In carnivores, the genera Eimeria and Cystoisospora and nematodes from Ancylostomatidae, Strongyloidae, Ascaridae, Capillariidae and Trichocephalidae were identified. Of 13 omnivore species included in the study, parasites from Eimeridae, Ascaridae, Strongyloidae, and Trichocephalidae were identified in seven species. Toxoplasma antibodies were identified using enzyme-linked immunosorbent assay in all definitive hosts (lions and wild cats) examined. In intermediate hosts (herbivores and omnivores), antibodies to Toxoplasma gondii had a prevalence of 58.8%, except in wild boars (Sus scrofa), in which the prevalence was 100%.

Evaluation of the health, prevalence, and incidence of diseases in wild populations is a critical component of wildlife management. In addition, the establishment of reference physiologic parameters can contribute to the assessment of population risks. Complete medical evaluations were performed on 33 wild Verreaux's sifaka (Propithecus verreauxi) from Kirindy Mitea National Park, Madagascar, during two field captures in June 2010 and June 2011. Each animal received a complete physical examination: weight, body temperature, heart rate, and respiratory rate were recorded, and ectoparasites collected. Blood samples were collected for complete blood cell count, differential white blood cell count, hemoparasite examination, serum biochemical profile, fat-soluble vitamin analysis, trace mineral analysis, and toxoplasmosis and viral serology. Fecal samples were collected for bacterial culture and endoparasite examination. Significant differences exist between age classes for neutrophil and lymphocyte count, alkaline phosphatase, and creatinine values. Parasites detected were Callistoura sp., Bertiella lemuriformis, and Dipetalonema petteri. This publication reports the first complete biomedical evaluation of the P. verreauxi and provides a basis for hematologic and biochemical comparisons of P. verreauxi in the wild.

Point of care devices can assess electrolyte, blood gas, biochemical, and hematologic values in a critical care setting. Although these devices are commonly used in humans and companion mammals, few studies have assessed their use in avian species. This study compares electrolyte, hemoglobin (Hgb), hematocrit (Hct), acid-base, and venous blood gas parameters between the i-STAT® and IRMA TruPoint® blood gas analysis systems for 35 Quaker parrots. Agreement between the two analyzers and the effect of gender, time lag between sample analysis, and cartridge expiration were evaluated. Male birds had increased Hgb and Hct compared with females, independent of analyzer method. In expired i-STAT cartridges, only glucose significantly increased. Packed cell volume determined by centrifugation was higher than Hct, as calculated by either analyzer. The analyzers had good agreement for total carbon dioxide, bicarbonate, pH, and Hgb, fair agreement for potassium (K), ionized calcium (iCa), venous partial pressure of carbon dioxide, and base excess, and poor agreement for sodium (Na), venous partial pressure of oxygen (PO₂), and oxygen saturation (SO₂). Values for Na, iCa, PO₂, and SO₂ were significantly higher on the IRMA than the i-STAT, while K was significantly lower on the IRMA when compared with the i-STAT. The time lag between sample analyses on the i-STAT and IRMA did not be correlate to any analyte changes. Despite these differences, both the i-STAT and the IRMA appear to be acceptable clinical tools in avian critical care, although reference ranges for each analyzer should be created.

It was observed previously that end-expired carbon dioxide (P_ÉCO₂) decreased when immobilized black rhinoceroses (Diceros bicornis) were moved from sternal to lateral recumbency. These experiments were designed to test whether greater alveolar ventilation or greater pulmonary dead space in lateral recumbency explains this postural difference in P_ÉCO₂. Twenty-one (9 male, 12 female; 15 [3.5–26] yr old) wild black rhinoceroses were immobilized with etorphine and azaperone and positioned in either sternal or lateral recumbency. All rhinoceroses were hypoxemic and had lactic and respiratory acidemia. The animals in lateral recumbency were more acidemic, had higher lactate, and lower arterial oxygen that those in sternal recumbency; however, arterial carbon dioxide was similar between groups. Both P_ÉCO₂ and mixed expired carbon dioxide pressure were lower in lateral than sternal recumbency. Although there was no difference in tidal volume or arterial carbon dioxide, both the breathing rate and minute ventilation were greater in lateral recumbency. The physiologic dead space ratio and dead space volume were approximately two times larger in lateral recumbency; hence, the decrease in P_ÉCO₂ in lateral recumbency can be attributed to increased dead space ventilation not increased alveolar ventilation. Positioning immobilized rhinoceroses in lateral recumbency does not confer any advantage over sternal in terms of ventilation, and the increase in minute ventilation in lateral recumbency can be considered an energetic waste. Although arterial oxygen was superior in sternal recumbency, further studies that measure oxygen delivery (e.g., to the muscles of locomotion) are warranted before advice regarding the optimal position for immobilized rhinoceroses can be given with confidence.

For treatment of hypoxemia, delivery of the minimum effective oxygen flow rate is advantageous during field anesthesia because it prolongs the life of the oxygen cylinder. Portable oxygen concentrators as the oxygen source require less logistical considerations than cylinders and are a safer alternative during helicopter field work because they are nonexplosive devices. The objective of this study was to evaluate low oxygen flow rates by continuous or pulsed intranasal delivery for treatment of hypoxemia in anesthetized white-tailed deer (Odocoileus virginianus). Nine captive adult female deer (body mass 56–72 kg) were physically restrained in a drop-floor chute and hand injected intramuscularly with medetomidine (0.1–0.14 mg/kg) and ketamine (2.5–4.3 mg/kg). Intranasal oxygen was delivered from an oxygen cylinder at continuous flow rates of 1 and 2 L/min or from a battery driven oxygen concentrator (EverGo^TM Portable Oxygen Concentrator, Respironics®) with pulse-dose delivery (maximum capacity of 1.05 L/min). The pulse-dose setting (pulse volume 12–70 ml) was adjusted according to the respiratory rate. Arterial blood gases were analyzed before, during, and after O₂ supplementation. A 10-min washout period was allowed between treatment groups. All three treatments adequately treated hypoxemia. The partial pressure of arterial oxygenation increased significantly from baseline values of 55 ± 10 to 115 ± 31 mm Hg during supplementation from the oxygen concentrator, to 138 ± 21 mm Hg during supplementation from the oxygen cylinder at 1 L/min, and to 201 ± 42 mm Hg at 2 L/min. In conclusion, low flow rates of intranasal oxygen supplemented continuously from an oxygen cylinder or by pulsed delivery from a portable oxygen concentrator effectively treated hypoxemia in anesthetized white-tailed deer.

The yellow-marginated box turtle (Cuora flavomarginata) is critically endangered and very few studies have been performed to aid the conservation effort. This study reports the first complete analysis of blood parameters for this species and will provide a reference for future conservation studies. Hematology and biochemistry reference range values were established from 52 (18 males and 35 females) clinically healthy yellow-marginated box turtles (C. flavomarginata). In order to investigate the gender and seasonal differences in these ranges, blood samples were collected from the jugular veins of the turtles in January, April, July, and October to represent the four seasons. To detect significant variation (P < 0.05), data were analyzed using repeated measurements of analysis of variance. Significant gender differences were observed in red blood cell mass parameters, absolute eosinophils, absolute monocytes, alkaline phosphatase, aspartate aminotransferase, blood urea nitrogen, calcium, uric acid, cholesterol, creatine kinase, glucose, phosphorus (P), total protein, and triglycerides. Marked seasonal variations were noted in red blood cell mass parameters, absolute monocytes, albumin, alkaline phosphatase, aspartate aminotransferase, blood urea nitrogen, uric acid, creatine kinase, creatinine, glucose, P, and triglycerides. Primary reasons for the differences between gender and season were associated with reproduction and temperature change. Cuora flavomarginata has lower alanine aminotransferase values compared with other chelonians.

In the years 2011–2012, a consensus nested polymerase chain reaction was used for the detection of adenovirus (AdV) infection in reptiles. During this screening, three new AdVs were detected. One of these viruses was detected in three lizards from a group of green striped tree dragons (Japalura splendida). Another was detected in a green anole (Anolis carolinensis). A third virus was detected in a Jackson's chameleon (Chamaeleo jacksonii). Analysis of a portion of the DNA-dependent DNA polymerase genes of each of these viruses revealed that they all were different from one another and from all previously described reptilian AdVs. Phylogenetic analysis of the partial DNA polymerase gene sequence showed that all newly detected viruses clustered within the genus Atadenovirus. This is the first description of AdVs in these lizard species.

Ranavirus is an emerging disease that infects fish, amphibians, and reptiles. Ranavirus induces an inflammatory response leading to death in many susceptible species. Red-eared sliders (RES; Trachemys scripta elegans) are vulnerable to ranavirus infection and are economically significant chelonians kept in the pet trade and utilized in research. Early identification of RES with inflammatory diseases would allow for isolation of affected individuals and subsequent disease investigation, including molecular testing for ranavirus. Validation of an inexpensive, clinically relevant, and reproducible diagnostic test that detects inflammation in turtles is needed. Although commonly used, plasma protein electrophoresis to detect an inflammatory acute-phase protein response has not been evaluated in a controlled environment in turtles with experimentally induced inflammatory disease. The objective of this study was to measure plasma protein fractions by electrophoresis to determine if an acute-phase protein response occurs in RES during infection with a frog virus 3–like ranavirus (FV3-like virus) isolated from a chelonian. A Bradford assay and agarose gel electrophoresis (AGE) were performed using plasma collected during a study of the effect of temperature on the pathogenesis of ranavirus in RES. In RES at the time of viremia, total albumin (ALB_mg/ml) and albumin to globulin ratio were significantly lower and β-globulin percentage was significantly higher in RES exposed to ranavirus (n = 4) as compared to matched, uninfected RES (n = 8). In the last sample collected prior to death, total protein (TP_mg/ml), ALB_mg/ml, α-globulin percentage, and total α-globulin (α_mg/ml) were significantly lower in RES exposed to ranavirus (n = 4) than control individuals (n = 8). In summary, FV3-like virus induces a decrease in plasma albumin concentration at the onset of viremia and decreases in TP_mg/ml, ALB_mg/ml, and α_mg/ml concentrations prior to death in RES as measured by AGE.

Five acute-phase reactants—serum amyloid A (SAA), C-reactive protein (CRP), haptoglobin, albumin, and iron—were measured using commercially available assays in 110 healthy rhesus macaques (Macaca mulatta), and reference intervals were established for future use in health monitoring of this species. Reference intervals established were as follows: SAA, 29.5–87.7 mg/L; CRP, 0–17.5 mg/L; haptoglobin, 354.3–2,414.7 mg/L; albumin, 36.1–53.0 g/L; and iron, 13.3–40.2 μmol/L. Furthermore, changes in the acute-phase reactants were studied in two additional groups of animals: eight rhesus macaques suffering from acute traumatic injuries and nine rhesus macaques experimentally infected with Mycobacterium tuberculosis reflecting a chronic active inflammation. In animals with inflammation, SAA and haptoglobin concentrations were moderately increased, while CRP increased more than 200-fold. In addition, marked decreases in albumin and iron concentrations were observed. These results show that SAA, CRP, and haptoglobin are positive acute-phase proteins, whereas albumin and iron are negative acute-phase reactants in rhesus macaques.

Captive Eld's deer (Rucervus eldi thamin) were evaluated for the presence of rabies virus–neutralizing antibodies using a rapid fluorescent focus inhibition after vaccination with either a live canarypox-vectored recombinant rabies vaccine or a killed monovalent rabies vaccine. Twelve deer were vaccinated with 1.0 ml of killed, adjuvanted, monovalent rabies vaccine at 5–33 mo of age then annually thereafter, and 14 deer were vaccinated with 1.0 ml nonadjuvanted, live canarypox-vectored rabies vaccine at 3–15 mo of age then annually thereafter. Banked serum was available or collected prospectively from deer at 6 mo and 1 yr after initial vaccination, then collected annually. Rabies virus–neutralizing antibodies considered adequate (>0.5 IU/ml) were present in 20/34 samples vaccinated with canarypox-vectored rabies vaccine and in 12/14 samples vaccinated with killed adjuvanted rabies vaccine. Poor seroconversion was noted in deer less than 6 mo of age vaccinated with the canarypox-vectored rabies vaccine.

Cardiac disease is a common finding in small mammals but it is rarely reported in striped skunks (Mephitis mephitis). The aim of this survey was to evaluate the prevalence of cardiac disease in striped skunks and to characterize the types of cardiac disease that might be present. In April 2010, a questionnaire was sent to veterinarians in zoologic collections with membership in the International Species Inventory System. Surveys were distributed to 55 institutions in the United States, Canada, and Europe. Twenty collections with a total of 95 skunks replied to the questionnaire. Of these, five collections reported at least one skunk with cardiac conditions for a total of 11 cases. In these 11 animals, the following conditions were diagnosed: myocardial fibrosis (n = 4), myxomatous valve degeneration (n = 4), hypertrophic cardiomyopathy (n = 1), dilated cardiomyopathy (n = 1), and valvular endocarditis (n = 1). Based on these findings, cardiac diseases should be considered as part of the differential diagnosis in captive striped skunks presenting with weakness, lethargy, and decreased appetite. Cardiac ultrasound also should be considered at the time of annual health examinations to evaluate for possible cardiac conditions at an early stage.

A prospective study to assess changes in selected plasma biochemistry and electrolyte values, plasma insulin and aldosterone concentrations, and electrocardiography (ECG) was performed on eight female captive tigers (Panthera tigris) and three lions (Panthera leo) undergoing general anesthesia for elective laparoscopic ovariectomy. Each animal was sedated with medetomidine (18–25 μg/kg) and midazolam (0.06–0.1 mg/kg) intramuscularly, and anesthesia was induced with ketamine (1.9–3.5 mg/kg) intramuscularly and maintained with isoflurane. Venous blood samples were collected and analyzed for plasma biochemistry parameters and insulin and aldosterone concentrations. An ECG was recorded at the time of each blood sample collection. Mean plasma potassium, glucose, phosphorus, and aldosterone concentrations increased during anesthesia (P ≤ 0.05). One tiger developed hyperkalemia (6.5 mmol/L) 2.5 hr after anesthetic induction. Plasma insulin concentrations were initially below the low end of the domestic cat reference interval (72–583 pmol/L), but mean insulin concentration increased (P ≤ 0.05) over time compared with the baseline values. Three tigers and two lions had ECG changes that were representative of myocardial hypoxemia. Based on these results, continuous monitoring of clinical and biochemical alterations during general anesthesia in large nondomestic felids is warranted, and consideration should be given to reversal of medetomidine in these animals should significant changes in electrolytes or ECG occur.

Crenosoma striatum is a species of parasitic nematodes from the family Crenosomatidae responsible for pathologic lung lesions in the hedgehog (Erinaceus europaeus). Infection with C. striatum can cause weight loss, dry cough, and bronchitis. In the present study, hedgehogs killed by road accidents, or trapped and found dead on farms in different parts of Mazandaran province (Iran), were transferred to the laboratory. After dissection, parasite samples collected from the lung were placed into 70% alcohol. After clarification with lactophenol and subsequent staining, the nematodes were identified as C. striatum according to previously published guidelines. For histopathologic examination, lung samples were collected. The tissues were fixed and following routine processing, sections were stained with hematoxylin and eosin. Microscopic diagnoses included hyperemia, eosinophilic bronchointerstitial pneumonia, thickening of the interstitium, and eosinophilic microabscesses in bronchial airways. Eosinophilic pneumonia was characterized by eosinophil and other mononuclear leukocyte infiltration within the lung interstitium. Crenosoma striatum can lead to mortality in hedgehogs.

Iron overload is common in lemurs and some New World nonhuman primates raised in captivity, but there is no such documentation in the red bald-headed uakari (Cacajao calvus rubicundus). This study describes postmortem documentation of severe iron storage disease in one red bald-headed uakari and the use of iron chelation with oral deferasirox in the three surviving members of the colony. Magnetic resonance imaging was used to quantify pretreatment iron burden and to follow the response to therapy in two females, 22 and 28 yr of age, and one male 33 yr of age. Baseline liver iron concentrations ranged from 16 to 23 mg/g dry weight. In humans, a liver iron concentration greater than 15 mg/g is considered severe and associated with endocrine and cardiac toxicity. The uakaris were otherwise asymptomatic, generally healthy, nonpregnant, and on a stable, low-iron diet. Quantitative magnetic resonance imaging indicated that dosage escalations up to 100 mg/kg were needed to produce meaningful reductions in iron stores. After 5 yr of therapy, two animals continue at a dosage of 100 mg/kg per day, and the third was transitioned to twice-weekly maintenance dosing because of successful de-ironing. The animals tolerated iron chelation therapy well, having stable hematologic, renal, and hepatic function profiles before, during, and after treatment. Deferasirox monotherapy may represent a therapeutic option in primates with iron storage disease when dietary measures are ineffective and phlebotomy is logistically challenging.

Recent publications report that some therapeutic drugs used in koalas (Phascolarctos cinereus) have poor oral absorption and are rapidly eliminated. Therefore, information on both the analgesic drug dosage regimens used to treat koalas in Australia and koala caretakers' perceptions of the efficacy of these drugs to control pain was collected for the purpose of identifying the most popular analgesics to prioritize future analgesic pharmacokinetic studies for this species. A one-page, double-sided questionnaire was distributed both electronically and by mail to Australian koala care facilities such as zoos and wildlife hospitals. Information was received from 13 respondents. Nonsteroidal anti-inflammatory drugs (NSAIDs) were the most frequently used analgesics, followed by full μ- and partial opioid receptor agonists and acetaminophen with or without codeine. The full μ-opioid receptor agonists and acetaminophen with or without codeine were most consistently considered efficacious, with wider variation in perceived efficacy of the NSAIDs. Analgesic drug combinations were generally thought efficacious.

A loggerhead sea turtle (Caretta caretta) was found stranded alive along the Adriatic coast close to Ancona, Italy, displaying obtundation, tachypnea, and increased respiratory effort. It died a few hours after admission, and a postmortem examination was immediately performed. Miliary yellowish nodules were evident in the liver, and a lower number in the heart, stomach, and gut wall. Hundreds of whitish nodules were scattered in the lungs, with the majority of the pulmonary parenchyma being replaced by the lesions. Histologically, all nodular lesions consisted of a small central area of necrosis with acid-fast bacilli surrounded by epithelioid cells, macrophages, and lymphocytes. Giant cells were found in the spleen and the liver. Kidneys, lungs, liver, spleen, brain, and skin lesions were inoculated aseptically onto general isolation media and selective isolation media for mycobacteria. The isolate showed a restriction pattern identical to Mycobacterium chelonae by polymerase chain reaction–restriction fragment length polymorphism. To the best of the authors' knowledge, this is the first description of a disseminated infection caused by a potentially pathogenic mycobacteria in a stranded, free-ranging loggerhead sea turtle. Veterinary staff and biologists who handle sea turtles with suspected mycobacterial disease should protect themselves appropriately.

Used since the 1970s as an avian anesthetic, the neurosteroid alfaxalone has been reformulated to avoid side effects from its castor oil excipient. This case report describes the clinical use of a new alfaxalone formulation (Alfaxan) as an intravenous anesthetic induction agent in wild isoflurane-anesthetized rose flamingos (Phoenicopterus roseus). Twenty-five male and female rose flamingos underwent orthopedic surgery using isoflurane anesthesia. The animals were induced following one of two protocols: inhaled isoflurane by facemask (ISO; n = 9) or intravenous alfaxalone (2 mg/kg; ALF; n = 16). The time and quality of anesthetic induction (until first signs of muscle relaxation) and the time and quality of recovery (sternal recumbency) were recorded using a scoring system. Mild sedation was first observed at 18.4 ± 3.8 min and 1.7 ± 0.3 min, following isoflurane and alfaxalone administration, respectively (P < 0.001). Alfaxalone induction time was significantly shorter and induction quality was considered smoother than in the ISO group. Flamingos given alfaxalone induction required lower isoflurane concentrations for maintenance anesthesia than did flamingos induced with mask isoflurane (1.5–2 %vol vs. 4–5 %vol for ALF vs. ISO, respectively). Alfaxalone produced moderate cardiorespiratory effects not seen in the isoflurane induction group. Recovery times were similar with both protocols without significant differences in quality and length. The new alfaxalone formulation produces a safe and effective anesthetic induction in rose flamingos and has significant isoflurane-sparing effects during anesthesia.

Two captive adult female Tasmanian devils (Sarcophilus harrisii) were investigated for pruritis and dermatitis. In both cases skin lesions consisted of multifocal, superficial patches of crusting, hyperkeratosis, and ulceration. Lesions started on the ventral surfaces of the animal but then appeared on the dorsum as the disease progressed. In both animals, a diagnosis of cutaneous T-cell lymphoma was made based on histologic appearance of skin biopsies using immunohistochemistry. Attempt at treatment with lomustine 20 mg p.o. once every 3 wk in one individual did not slow progression of the condition. As a result of their propensity for developing neoplastic conditions, the use of chemotherapeutic agents in Tasmanian devils warrants further investigation.

Since it is reported to be difficult to establish Asiatic golden cat (Catopuma temmincki) breeding pairs in captivity as a result of overaggressive behavior of the male, artificial insemination (AI) may be a desired option by which to achieve pregnancy. This approach was chosen in the present case involving a nulliparous, 6-yr-old female cat that was inseminated transcervically during a naturally occurring estrus, which therefore required only a single general anesthetic procedure. On day 4 of estrus behavior, the male was anesthetized and semen was collected via urethral catheterization (UC) to recover spermatozoa in high concentration followed by electro-ejaculation (EE) to obtain additional semen and seminal fluid. The fresh UC semen, totaling 180 μl in volume and containing spermatozoa showing 55–70% sperm motility, was inseminated 2.5 hr later via a commercial cat urinary catheter passed through the cervix into the uterus. Immediately afterwards, the EE fraction (100 μl) was inseminated deeply into the dorsal medial fold of the vagina. The GnRH analogue Receptal® (0.75 ml, i.m.) was given during anesthesia in an attempt to induce ovulation. Increasing fecal concentrations of progesterone after AI and a significant rise in fecal prostaglandin F2α metabolite (PGFM) concentrations (P < 0.0001) from day 45 post-AI indicated that the cat had conceived, and it produced healthy twin cubs after an 84-day gestation.

Hemangiosarcoma, hemangioma, transitional cell carcinoma, and chronic cystitis were diagnosed in the urinary bladder of six captive fallow deer (Dama dama). Hematuria and thin body condition were observed in the advanced cases. These findings were compatible with chronic enzootic hematuria and were suspected to have been induced by chronic ingestion of bracken fern (Pteridium aquilinum) present on the premises. These lesions were similar to those described in bovine enzootic hematuria in cattle. Hemangiosarcoma metastases, hydronephrosis, and renal carcinoma were also associated in some cases. This is the first report of hemangioma, hemangiosarcoma, transitional cell carcinoma, and renal carcinoma of the urinary bladder in fallow deer and the first indication of bracken fern intoxication in deer.

A 32-day-old, hand-reared, captive-born female clouded leopard (Neofelis nebulosa) cub presented as being unable to stand, ambulate, or adduct both hind limbs. The cub exhibited hyperextension of both tarsal joints and a flattened thorax, which limited mobility to “swimmer-like” movements. Neither congenital defects nor neurologic deficits were observed during the medical examination. Radiographic examination showed the thorax was compressed dorsoventrally, but no other skeletal abnormalities were detected. Based on clinical signs, the condition was more consistent with swimmer syndrome, which has been described in young offspring of several domestic species. Over the course of 3 wk, affected limbs were treated by intensive physiotherapy, corrective bandages were applied, and thermotherapy was used to improve circulation, which resulted in a complete recovery and development of subsequent normal ambulation. It is concluded that early diagnosis and treatment of this condition led to the resolution of clinical signs, resulting in normal development of the clouded leopard cub reported here.

Cefovecin was administered to six healthy adult white bamboo sharks (Chiloscyllium plagiosum) and six healthy adult Atlantic horseshoe crabs (Limulus polyphemus) to determine its pharmacokinetics in these species. A single dose of cefovecin at 8 mg/kg was administered subcutaneously in the epaxial region of the bamboo sharks and in the proximal articulation of the lateral leg of the horseshoe crabs. Blood and hemolymph samples were collected at various time points from bamboo sharks and Atlantic horseshoe crabs. High performance liquid chromatography was performed to determine plasma levels of cefovecin. The terminal half-life of cefovecin in Atlantic horseshoe crabs was 37.70 ± 9.04 hr and in white bamboo sharks was 2.02 ± 4.62 hr. Cefovecin concentrations were detected for 4 days in white bamboo sharks and for 14 days in Atlantic horseshoe crabs. No adverse effects associated with cefovecin administration were seen in either species.

Multiple organ invasion by keratinophilic fungi in the green iguana (Iguana iguana) has not been previously reported. In this case, a 1-yr-old female green iguana presented with a nodular, darkly discolored skin lesion surrounded by necrosis in the right ventral abdominal region. A cytologic examination of the fine needle aspiration of the lesion revealed an exuberant proliferation of fibroblasts, macrophages, and multinucleated cells along with frequent filamentous structures consistent with hyphal elements. The necropsy revealed diffuse infiltration of the liver, lung, and cardiac apex with white nodules. A histopathologic examination of the lesions also confirmed a fungal infection associated with granulomatous inflammation. Rapid polymerase chain reaction (PCR) analysis of the chitin synthase 1 gene was conducted for rapid direct detection, and inter-simple sequence repeat fingerprinting was conducted to classify the infectious origin. The PCR analysis definitively demonstrated representative Microsporum canis fungus. The present report is the first case of disseminated M. canis infection with multiorgan involvement in a green iguana.

Contraception is necessary to manage zoo animal populations and to be able to house animals in groups without producing additional unwanted offspring. In felids and canids, an association between exposure to progestins and the occurrence of endometrial and mammary gland pathology has been documented. Therefore, the Association of Zoos and Aquariums (AZA) Wildlife Contraceptive Center recommends the use of deslorelin acetate for long-term contraception in carnivores. Return to cyclicity after deslorelin treatment has been variable; some individuals show ovarian suppression for long periods after the expected end of the deslorelin efficacy. In an attempt to reduce the time to reversal, techniques to locate and remove previous implants are being developed. This report documents the successful implementation of high-frequency ultrasonography in lions (Panthera leo) to locate and direct surgical removal of multiple deslorelin implants placed at least 2 yr previously as well as the return of follicular activity in both females at 7 months post-removal of implants.

A 13-yr-old female two-toed sloth (Choloepus didactylus) with a prolonged history of worsening azotemia was necropsied shortly after euthanasia. On necropsy, the sloth had poor body condition, bilaterally shrunken kidneys, and a large neoplastic mass replacing the right liver lobe. Histologic examination demonstrated chronic renal disease with metastatic mineralization as the cause of morbidity. The liver mass was not associated with any known clinical signs and was diagnosed as a solitary and well-differentiated hepatocellular carcinoma. To the authors' knowledge, this is the first report of hepatocellular carcinoma diagnosed in a sloth and the first detailed description of chronic renal disease in this species.

A 59-yr-old, captive female African elephant (Loxodonta africana) died of a cardiovascular collapse. Necropsy revealed bilateral replacement of the ovarian tissue by multiple cystic formations of up to 10 cm in diameter. The cysts were thin walled and filled with clear watery fluid. Smaller solid masses with an irregular surface projected from the wall of a few of the cysts. Histologically, the cystic structures were characterized by a single layer of well-differentiated cuboidal epithelial cells resting on a basement membrane. Occasionally the cysts contained nodular proliferations of single-layered, well-differentiated cuboidal epithelial cells forming anastomosing tubules and occasional papillary projections. The ovarian neoplasia was diagnosed as a bilateral multilocular serous ovarian cystadenoma. The nulliparous status and the advanced age may have contributed to the ovarian pathology.

Pseudomonas aeruginosa is an important pathogen that has been implicated in a number of serious conditions in humans and animals. Studies regarding the efficacy of antimicrobial drugs against virulent strains of P. aeruginosa in human and animal hosts have demonstrated that common therapeutic options are sometimes ineffective. In captive animals, in particular reptiles, literature on this topic is scarce. In this study, a 6-yr-old monitor lizard, Varanus niloticus, died suddenly without previous symptoms. Postmortem examination and bacteriologic analysis of necropsy samples revealed severe fibrinonecrotic enteritis caused by P. aeruginosa. The isolate presented a typical resistance profile, showing resistance to amoxycillin-clavulanic acid, ampicillin, cephotaxime, and sulphamethoxazole-trimethoprim. Virulence characterization demonstrated that the bacterium was able to express hemolysins, DNases, gelatinases, and lipases and had biofilm-forming ability. This study highlights the importance of surveillance for antimicrobial resistance and virulence traits in captive animals, which may present a challenge to practitioners.

An 8-yr-old female slender-tailed meerkat (Suricata suricatta) presented with a necrotic sublingual mass and osteolysis of the mandible. After 1 mo of palliative care, the meerkat was euthanized. The mass was diagnosed histologically as an anaplastic carcinoma with extensive rostral mandibular destruction. Immunohistochemistry for vimentin and cytokeratin was validated in this nontypical species and showed that neoplastic cells expressed both mesenchymal and epithelial characteristics, suggestive of a primitive and poorly differentiated tumor. A review of 150 adult slender-tailed meerkat histopathology reports showed a 2% prevalence of orofacial neoplasia, suggesting that oral neoplasms are uncommon in meerkats.

A 16-yr-old polar bear (Ursus maritimus) presented with severe diarrhea shortly following transfer to the North Carolina Zoological Park. Multiple diagnostic procedures were performed over several months and the cause of the chronic diarrhea was inconclusive. Histologically, colonic mucosal biopsies were consistent with severe chronic eosinophilic and lymphoplasmacytic colitis with no evidence of etiologic agents present. A dietary elimination trial was conducted and an adverse food reaction to the dog chow in the diet was confirmed.

A female southern white rhinoceros (Ceratotherium simum simum) calf presented with a patent urachus at 5 days of age. It was immobilized for examination and medical treatment was elected over surgical repair. Systemic antibiotics and topical treatment of the site by cleaning with chlorhexidine solution were followed by silver nitrate cautery. This animal required no further treatment and has reached 1 yr of age with no other medical problems. Although patent urachus is relatively common in domestic animals, especially horses, neither the problem nor its resolution are well documented in exotic animals and surgical intervention is more commonly performed.

A 42-year-old female Indian elephant (Elephas maximus indicus) developed a sudden onset of excessive salivation and dysphagia. Esophageal obstruction was suspected; possibly related to palm frond ingestion. Esophageal endoscopy revealed a mat of plant material in the distal esophagus. An initial attempt at relieving the obstruction was unsuccessful, but subsequent use of custom-made instruments along with insufflation and hydropulsion enabled partial removal of the material. Postimmobilization care included aggressive intravenous and rectal fluids, anti-inflammatory and antibiotic administration, and fasting. Despite treatment, the dysphagia persisted and the elephant was euthanized due to lack of improvement and grave prognosis. Postmortem examination revealed remaining plant material in the esophagus, complicated by an esophageal dissection, mural hematoma, and secondary bacterial infection. Iatrogenic trauma may have contributed to the extent of esophageal injury. Although treatment was ultimately unsuccessful, the supportive care employed could potentially aid recovery in cases of less severe esophageal trauma.

A subadult female loggerhead sea turtle (Caretta caretta) was caught in a trawl net off the west coast of Florida with a spotted eagle ray (Aetobatus narinari) spine lodged in the left stifle. Surgical removal of the spine was performed and antibiotic treatment was initiated. Four weeks later, endoscopy revealed a second spine entering an intestinal lumen. The fistulous tract of the left prefemoral fossa was surgically excised and the intestinal perforation was repaired. Dehiscence occurred and a vacuum-assisted closure (VAC) system was used on the wound for approximately 18 days to help reduce infection and increase the rate of healing. The left stifle wound was treated to heal by second intention. The turtle remained in rehabilitation for 19 mo before being released off the west coast of Florida. This case describes stingray envenomation injuries as a complex and potentially life-threatening bycatch effect to sea turtles caught in trawl nets.

Orthoreoviruses have been associated with disease in reptiles, but have not previously been isolated from snakes with inclusion body disease (IBD). An orthoreovirus was isolated from a Boa constrictor diagnosed with IBD and then used to conduct a transmission study to determine the clinical importance of this virus. For the transmission study, 10 juvenile boas were experimentally infected with the isolated orthoreovirus and compared to 5 sham-infected control animals. Orthoreovirus was reisolated for a period of 18 wk after infection and weight gain was reduced in infected snakes. Histological examination showed a mild hepatitis in three of four virologically positive snakes up to 12 wk after infection. Results indicated that the orthoreovirus was moderately pathogenic, but, no evidence was found to indicate that it was the causal agent of IBD. In the light of the discovery of Arenaviruses in some snakes with IBD, it was proposed that orthoreoviruses may play a role in synergistic infection.

A 16-yr-old northern water snake (Nerodia sipedon) presented with a large, firm midcoelomic swelling. A complete blood count, survey radiographs, coelomic ultrasound, and fine needle aspirate of the mass were performed. Survey radiographs revealed a soft tissue opacity mass. Ultrasonographic examination confirmed the presence of the mass but could not aid in its visceral localization. After 2 weeks, the snake presented again because of continued anorexia and poor quality of life. Euthanasia was performed. Gross necropsy revealed a multilobulated mass attached to and effacing the splenopancreas. Histologically, the mass was composed of cuboidal to columnar neoplastic epithelial cells forming tubules surrounded by variable amounts of fibrovascular stroma. Histological examination and immunohistochemical staining of other tissues revealed local invasion in the subserosa and tunica muscularis of the stomach, metastasis within the liver, in the mesovarium, and an intravascular metastasis within the ventricle of the heart surrounded by a thrombus.

A 25-yr-old, nulliparous, female Przewalski's wild horse (Equus ferus przewalskii) with a history of pituitary pars intermedia dysfunction and recent onset of serosanguinous vaginal discharge was euthanized after a period of lethargy and inappetance. Postmortem examination confirmed an infiltrative uterine adenocarcinoma, which is an uncommon neoplasia in equids. Reproductive disease is significant in this species as they are considered endangered by IUCN. Reproductive soundness and success are paramount to conservation efforts.

A 13-yr-old male African black-footed penguin (Spheniscus demersus) presented thrice over 7 mo with gastrointestinal obstruction secondary to cloacolithiasis. Clinical signs consistently resolved with cloacolith removal and supportive care. However, 10 mo after initial presentation, it presented with similar signs, plus significant weight loss. No cloacolith was found, and it subsequently died. Significant gross findings included bilateral cecal masses, colonic perforation, and marked secondary coelomitis, multifocal tan to pale hepatic nodules, and pale kidneys with miliary white foci. Histopathologic diagnoses were intestinal lymphosarcoma with hepatic and renal metastases, secondary intestinal rupture, and subacute severe bacterial coelomitis. To the authors' knowledge, this is the first full report of either cloacolithiasis or lymphosarcoma in a penguin.

Three koalas (Phascolarctos cinereus) presented with moderate to severe interproximal space accumulation with Eucalyptus spp. at the San Diego Zoo. Premise™ trimodal composite was used to create dental onlay bridge-like prostheses to eliminate open, enlarged interproximal spaces. The prostheses prevented further leaf material accumulation within the interproximal spaces and consequently reduced periodontal disease. Aesthetically, the prosthesis replicates normal tooth coloration and appearance and wears at a similar rate to surrounding teeth. Prosthetic repair or replacement may be required, so periodic examination every 3–6 mo is recommended.

A 3-mo-old red fox (Vulpes vulpes) developed generalized crusty plaques on its body during rehabilitation after an automobile accident requiring amputation of one leg. Histologic examination of skin lesion biopsy revealed granulomatous dermatitits with many intralesional protozoal tachyzoites. The protozoa stained positively with antibodies to Neospora caninum but not to Toxoplasma gondii. Treatment with clindamycin hydrochloride (10 mg/kg, twice daily, s.c.) for 1 mo completely resolved lesions, and protozoa were not demonstrable in biopsy of skin after treatment. The fox had agglutinating antibodies to T. gondii (modified agglutination test, titer 1 : 3200) and N. caninum (Neospora agglutination test, titer 1 : 25), and viable T. gondii (genotype III) was isolated from the skin biopsy after treatment. This is the first report of clinical neosporosis in a wild canid.

The first cases of infection with the eyeworm Thelazia callipaeda (Spirurida, Thelaziidae) in red foxes (Vulpes vulpes) from Portugal are described. Worms were collected from 1 fox (7 worms) in the north and from 2 foxes (10 worms) in the central region of the country. Partial molecular amplification of mitochondrial cythocrome c oxidase subunit 1 gene and sequencing revealed a 100% homology with T. callipaeda haplotype 1, which is the sequence type circulating in Europe. Data suggest that wildlife participate in maintaining this endemic infection in dog and cat populations from the studied areas. Furthermore, due to the zoonotic potential of T. callipaeda, there is a risk of infection in humans residing in areas where thelaziosis is present in wild and domestic animals.

A near–full-term alpaca (Vicugna pacos) was stillborn 2 days before the expected date of delivery; necropsy examination was conducted within 6 hr of delivery. Gross lesions were enlarged liver and hydrocephalus. On histologic examination, mild inflammatory lesions were identified in the placenta, liver, and lungs, although no etiology was recognized. Within the brain, there was a mild nonsuppurative meningoencephalitis, hydrocephalus, and hydromyelia. Both kidneys had inflammatory foci in cortex and medulla associated with protozoal tissue cysts. Protozoa in brain and kidneys were identified as Toxoplasma gondii based on immunoreactivity to T. gondii polyclonal antibodies that stain both tachyzoites and tissue cysts and BAG1 T. gondii antibodies that are bradyzoite specific. The tissue cysts exhibited intense positivity to T. gondii and mild immunoreactivity to Neospora caninum antibodies. The dam had a high antibody titer (1 : 12,800) to T. gondii and low titer (1 : 100) to N. caninum using their respective agglutination tests. This is the first report of toxoplasmosis-associated abortion in alpaca.