COMPARISON OF SUBCARAPACIAL SINUS AND BRACHIAL VEIN PHLEBOTOMY SITES FOR BLOOD COLLECTION IN FREE-RANGING GOPHER TORTOISES (GOPHERUS POLYPHEMUS).

Paired blood samples were collected from the brachial vein and subcarapacial sinus of 42 wild gopher tortoises (Gopherus polyphemus) for comparison of hematological and plasma biochemical, vitamin, mineral, and protein electrophoresis parameters with general linear models with repeated measures designs. Comparisons revealed higher brachial vein mean values for 38 of 42 parameters with significance (P ≤ 0.05) for 21 parameters. Significantly lower brachial vein means were noted for two parameters. Coefficients of variation for brachial vein samples were lower for 30 out of 42 parameters with significantly lower values for 11 parameters. Of the 12 parameters with lower coefficients of variation for subcarapacial sinus samples, significant differences were noted for four parameters. Both the lower coefficient of variation and the higher mean values for most parameters from brachial vein samples are attributed to greater hemodilution of subcarpacial samples with lymph compared to the paired brachial vein samples. To address the clinical relevance and reliability of the parameter ranges obtained from both paired phlebotomy sites, we compared these values to those obtained from other selected investigations of gopher tortoise and desert tortoise (Gopherus agassizii) clinicopathology. Although statistically significant differences were detected, available data demonstrated that most mean ± standard deviation values and/or ranges for both phlebotomy sites of our study are similar to those reported for the selected studies. Based on these results, utilization of either set of our study values in the assessment of a gopher tortoise population or individual gopher tortoises would be acceptable for clinical or management decisions. It is also advisable that investigators develop institutional blood parameter ranges for healthy individuals by phlebotomy site when possible in addition to maintaining standardized sample collection and handling protocols.

USE OF INTRACARPAL INTERLEUKIN RECEPTOR ANTAGONIST PROTEIN (IRAP) AND HYALURONIC ACID IN A MULTIMODAL THERAPEUTIC REGIME FOR OSTEOARTHRITIS IN AN ASIAN ELEPHANT (ELEPHAS MAXIMUS).

An approximately 41-yr-old female Asian elephant (Elephas maximus) experiencing forelimb stiffness and decreased range of motion was diagnosed with bilateral carpal osteoarthritis (OA). Standing sedation combined with local anesthesia was used to deliver ultrasound-guided carpal articular injections of an autologous conditioned serum product, interleukin receptor antagonist protein, combined with hyaluronic acid. Within 2 mo of completing therapy, improved range and speed of motion were evident. Reduced inflammation was suggested by decreased carpal articular prostaglandin E2 levels. Subjectively improved clinical signs lasted approximately 5-6 mo, at which point carpal articular injections were repeated. Joint inflammatory markers were useful in gauging response to treatment and may provide guidance in the diagnostic and therapeutic approach to elephant OA. On the basis of the positive response noted, interarticular autologous therapy combined with hyaluronic acid should be considered for carpal OA in elephants.

RETROSPECTIVE REVIEW OF OPHIDIOMYCOSIS (OPHIDIOMYCES OPHIODIICOLA) AT THE SMITHSONIAN'S NATIONAL ZOOLOGICAL PARK (1983-2017).

A retrospective review of systemic or localized mycotic infections in captive snakes confirmed via biopsy or necropsy from 1983 to 2017 was performed at the Smithsonian's National Zoological Park. Quantitative polymerase chain reaction (qPCR) confirmed infection with Ophidiomyces ophiodiicola (Oo) in 36.8% (n = 14) of the 38 mycotic infections. Infections with Oo were evenly distributed over the 35-y period and lacked a sex predilection. There was a period prevalence of 4.5% of completed snake necropsy or biopsy cases that were Oo positive. Species affected included green anaconda (Eunectes murinus, n = 4), garden tree boa (Corallus hortulanus, n = 1), false water cobra (Hydrodynastes gigas, n = 5), yellow anaconda (Eunectes notaeus, n = 1), eastern milksnake (Lampropeltis triangulum, n = 1), Brazilian rainbow boa (Epicrates cenchria cenchria, n = 1), and eastern diamondback rattlesnake (Crotalus adamanteus, n = 1). Histopathology demonstrated one or more of the following: heterophilic to necrotizing epidermitis with or without granulomatous dermatitis (n = 12), granulomatous pneumonia (n = 5), granulomatous endophthalmitis (n = 1), and subcutaneous-intramuscular fungal granuloma (n = 1). This study documents the presence of ophidiomycosis in a captive collection for almost 40 years, despite current literature designating it a recently emerging pathogen.

DERMATITIS AND RHINOSINUITIS CAUSED BY CURVULARIA SPECIES IN A CHINESE GORAL (NAEMORHEDUS GRISEUS).

Curvularia spp. are globally distributed saprophytic fungi, classified in the literature as dematiaceous, or darkly pigmented fungi. These fungi have been increasingly recognized as causing cutaneous, ocular, respiratory, and central nervous system infections in humans, but have been infrequently documented as pathogens in the veterinary literature. A 5-yr-old male Chinese goral (Naemorhedus griseus) presented with bilateral fungal dermatitis of the pinnae, and subsequent pyogranulomatous rhinosinusitis. Clinical signs included epistaxis, mucosanguineous nasal discharge, and dyspnea. Sequential histologic examinations of cutaneous and nasal lesions revealed pyogranulomatous inflammation with extracellular and phagocytized nonpigmented yeasts. Fungal culture and polymerase chain reaction identified Curvularia sp. The absence of pigmentation in tissue in this case suggests that pigmentation may not be a consistent histologic finding for this fungus, emphasizing the importance of molecular identification to prevent misidentification. Despite intensive interventions in this goral, the disease progressed, and was ultimately fatal.

BATRACHOCHYTRIUM DENDROBATIDIS IN A CAPTIVE COLLECTION OF GREEN SALAMANDERS ( ANEIDES AENEUS), LONG-TAILED SALAMANDERS ( EURYCEA LONGICAUDA), AND TWO-LINED SALAMANDERS ( EURYCEA BISLINEATA).

A chytridiomycosis outbreak from Batrachochytrium dendrobatidis ( Bd) in a mixed-species plethodontid salamander exhibit resulted in four green salamander ( Aneides aeneus) deaths. One green salamander died before treatment, and three died during treatment with daily 0.005% itraconazole baths. All salamanders had evidence of severe Bd infections via cytology, histopathology, and/or polymerase chain reaction (PCR) at the time of death. Ten long-tailed salamanders ( Eurycea longicauda) and one two-lined salamander ( Eurycea bislineata) that shared the enclosure were initially negative for Bd on quantitative PCR but were prophylactically treated with daily 0.01% itraconazole baths for 11 days. Posttreatment testing yielded eight long-tailed salamanders and one two-lined salamander positive for Bd with low gene equivalents. All salamanders were negative after two to three treatment courses, and there were no additional mortalities. The difference in mortality and fungal load suggested that genus Aneides salamanders may be more susceptible to Bd than genus Eurycea salamanders.

CALCINOSIS CIRCUMSCRIPTA IN A COHORT OF RELATED JUVENILE AFRICAN LIONS (PANTHERA LEO).

Three juvenile, genetically related African lions (Panthera leo) were evaluated for discrete dome-shaped subcutaneous masses present over the proximal lateral metatarsal-tarsal area. The lesions measured 3-8 cm in diameter, were fluctuant to firm, nonulcerated, and attached to underlying structures. On radiographic evaluation, the lesions were characterized by well-circumscribed punctate mineralizations in the soft tissue surrounded by soft tissue swelling without evidence of adjacent bony involvement. On cut surface, the lesions were made of numerous loculi containing 2-5-mm round-to-ovoid, white-to-gray, firm structures interspersed with fibrous tissue and pockets of serosanguinous fluid. Hematology, serum biochemistry, serum thyroid screening (including total thyroxine, total triiodothyronine, free thyroxine, and free triiodothyronine), and serum vitamin D panels (including parathyroid hormone, ionized calcium, and 25-hydroxyvitamin D) were unremarkable. Histopathologic evaluation of the lesions was consistent with calcinosis circumscripta with fibroplasia, chronic inflammation, and seroma formation. An additional two genetically related lions were considered suspect for calcinosis circumscripta based on presentation, exam findings, and similarity to the confirmed cases. All masses self-regressed and were not associated with additional clinical signs other than initial lameness in two cases.

Exploratory rigid laparoscopy in an African elephant (Loxodonta africana).

In March 2009, a 25-yr-old captive female African elephant (Loxodonta africana) underwent an exploratory laparoscopy after several weeks of diarrhea, submandibular and ventral edema, and swelling on medial and lateral aspects of all feet. Although there have been recent advances in laparoscopic vasectomies in free-ranging African elephants in South Africa utilizing specially designed rigid laparoscopes and insufflation devices, this was the first attempt at using these same techniques for an exploratory purpose. The elephant was sedated in a static restraint chute and remained standing for the duration of the procedure. Laparoscopy provided visibility of the dorsal abdomen, enabled collection of reproductive tract biopsies and peritoneal fluid samples, and allowed for instillation of antibiotics and crystalloid fluids directly into the abdominal cavity. Abdominal exploration, collection of tissue samples, and local therapy is possible via standing laparoscopy in megavertebrates.

Morbidity in a juvenile green sea turtle (Chelonia mydas) due to ocean-borne plastic.

An emaciated 2.36-kg juvenile green sea turtle, Chelonia mydas, was found floating off of Melbourne Beach, Florida, USA (28 degrees 2'4"N, 80 degrees 32'32"W). The turtle exhibited signs of cachexia, positive buoyancy, lethargy, and obstipation; was covered with barnacles; and was anorexic at the time of presentation. Dorsal-ventral radiographs with positive contrast confirmed obstruction of the gastrointestinal tract. Serum chemistry abnormalities reflected metabolic/nutritional deficiencies. Gastrointestinal prokinetics and oral/enema mineral oil applications were effective in relieving gastrointestinal obstruction with the turtle defecating a total of 74 foreign objects over a period of a month. After the removal of the foreign material, the turtle quickly regained normal behavior and health. The lack of blood parameters demonstrating infection or inflammation; the failure to respond to antibiotic and antifungal treatment as well as the parallel improvement in behavior and health after incremental evacuation of the plastic is highly suggestive of a cause and effect association.

Presumptive fenbendazole toxicosis in North American porcupines.

CASE DESCRIPTION: 4 North American porcupines were evaluated because of diarrhea or neutropenia (or both) that developed after treatment with fenbendazole for intestinal parasites. CLINICAL FINDINGS: Complete blood cell count abnormalities included severe neutropenia in all affected porcupines and mild anemia in some of them. In 2 porcupines, postmortem findings included bone marrow hypoplasia and intestinal crypt cell necrosis. TREATMENT AND OUTCOME: Affected porcupines received supportive care including fluid supplementation and broad-spectrum antimicrobials. The 2 surviving animals recovered after 9 to 33 days of treatment. CONCLUSIONS AND CLINICAL RELEVANCE: Fenbendazole is an anthelminthic that may be used in an extralabel manner for the treatment of intestinal parasitism in wildlife species. The drug inhibits mitosis and can affect rapidly dividing cell lines, such as those in the bone marrow and intestinal crypt mucosa. Fenbendazole may not be an appropriate anthelminthic choice in North American porcupines.

Hematologic and plasma biochemical changes associated with fenbendazole administration in Hermann's tortoises (testudo hermanni).

Toxicosis associated with benzimidazole anthelmintics has been reported with increasing frequency in zoologic collections. Clinical signs, clinicopathologic abnormalities, and gross and histologic lesions are primarily the result of damage to the gastrointestinal and hematopoietic systems. Profound leukopenia, especially granulocytopenia, is the most common and severe clinicopathologic change associated with benzimidazole administration. Death usually occurs from overwhelming systemic bacterial and/or fungal infections secondary to severe immunosuppression. In this 125-day study, six male Hermann's tortoises (Testudo hermanni) were treated orally with two 5-day courses of fenbendazole 2 wk apart at a dosage of 50 mg/kg. Serial blood samples were used to assess hematologic and plasma biochemical changes before, during, and following the treatment period. Although the tortoises remained healthy, blood sampling indicated an extended heteropenia with transient hypoglycemia, hyperuricemia, hyperphosphatemia, and equivocal hyperproteinemia/hyperglobulinemia, which were considered to be in response to fenbendazole administration. Changes in several other clinicopathologic parameters appeared to correlate with fenbendazole administration. The hematologic and biochemical changes seen in the healthy animals in this study should be considered when treating compromised tortoises with fenbendazole. Hematologic and plasma biochemical status of tortoises/reptiles should be determined before treatment and monitored during the treatment period. The risk of mortality of an individual from nematode infection should be assessed relative to the potential for metabolic alteration and secondary septicemia following damage to hematopoietic and gastrointestinal systems by fenbendazole.

Standing sedation in African elephants (Loxodonta africana) using detomidine-butorphanol combinations.

Standing sedation was provided for 14 clinical procedures in three African elephants (Loxodonta africana) managed by combined protected and modified-protected contact and trained through operant conditioning. An initial hand-injection of detomidine hydrochloride and butorphanol tartrate at a ratio of 1:1 on a microg:microg basis was administered intramuscularly, with a dosage range of 50-70 mg (12.9-19.7 microg/kg) for each drug. The initial injection resulted in adequate sedation for initiation and completion of eight procedures, whereas supplemental doses were required for the remaining procedures. The dosage range for the supplemental injections of each drug was 4.0-7.3 microg/kg. Initial effect was noted within 3.0-25 min (mean = 11.6 min, SD +/- 5.9 min), with maximal effect occurring at 25-30 min for those procedures not requiring supplementation. In all but one procedure, this effect was maintained until the end of the procedure, which ranged from 47 to 98 min (mean = 74.7 min, SD +/- 18.8 min). No cardiac or respiratory depression was appreciated. Recovery after administration of reversal agents was rapid and complete, ranging from 2 to 20 min (mean = 9.0 min, SD +/- 7.0 min). On the basis of the authors' experience, recommended dosage ranges for reversal agents would be intravenous yohimbine (73.4-98.5 microg/kg), intravenous naltrexone (48.9-98.5 microg/kg), and intramuscular naltrexone (73.4-98.5 microg/kg). Approximately one-third to one-half of the total naltrexone dose should be administered intravenously. Mild adverse side effects limited to the gastrointestinal tract were observed in association with five procedures including abdominal distention with or without transient anorexia. Administration of reversal agents, encouraging exercise and water consumption, and administration of flunixin meglumine were helpful in the resolution of signs. In addition to gastrointestinal signs, slight ataxia was observed before initiation of surgical stimulation during one procedure in which 19.7 microg/kg of each drug was administered. On the basis of the procedures that did not require supplementation to initiate treatment and taking into consideration the potential for ataxia at higher doses, a starting dosage range of 14.7-16.2 microg/kg of both detomidine and butorphanol in a ratio of 1:1 on a microg:microg basis administered i.m. simultaneously is recommended.

Bone marrow hypoplasia and intestinal crypt cell necrosis associated with fenbendazole administration in five painted storks.

Five painted storks were treated with fenbendazole for 5 days for internal parasitism. Four birds died following treatment. Profound heteropenia was a consistent finding in all samples evaluated; additionally, the 1 surviving bird had progressive anemia. Consistent necropsy findings in the 4 birds that died were small intestinal crypt cell necrosis and severe bone marrow depletion and necrosis. Fenbendazole has been associated with bone marrow hypoplasia and enteric damage in mammals and other species of birds. The dosages of fenbendazole used in birds are often substantially higher than those recommended for mammals, which may contribute to bone marrow hypoplasia and intestinal crypt cell necrosis associated with fenbendazole administration in birds.

Wound management in teleost fish: biology of the healing process, evaluation, and treatment.

Wound healing, although similar to terrestrial vertebrates, occurs faster in fish species in a proper aquatic environment. Wound management must involve identification and correction of potential noninfectious and/or infectious causes of disease. Wound evaluation includes assessment of the wound as well as systematic examination of the patient, water quality conditions, and husbandry practices. Diagnostic evaluation should, at a minimum, include physical examination, skin/cornea (if indicated) scraping, and lesions cultures. The clinician should also consider full thickness wound biopsies, gill and fin biopsies, blood collection, and radiographs for systemic evaluation as well. Local wound management should follow basic principles of wound care including cleaning and lavage, debridement, primary closure if indicated, and local and/or systemic antiinfectives. Monofilament absorbable or nonabsorbable sutures are commonly used for primary closure of wounds. Diagnostic results, patient and client compliance, and economic impact should guide systemic methods of treatment for wound care. Options for systemic therapy include oral or gavage, parenteral, bath, or prolonged immersion treatments. With bath or prolonged immersion therapies, aquatic systems must be properly prepared for treatment. Treatment of identified pathogens, proper water quality and husbandry, supportive care and nutrition must all be provided to promote an ideal wound healing environment for the teleost fish.

Mortality associated with melarsomine dihydrochloride administration in two North American river otters (Lontra canadensis) and a red panda (Ailurus fulgens fulgens).

Two adult North American river otters (Lontra canadensis) and an adult red panda (Ailurus fulgens fulgens) at three separate institutions died within 22 hr after receiving single 2.5- to 2.7-mg/kg doses of melarsomine dihydrochloride administered in the epaxial musculature as a treatment for filarid nematodes. One otter had a suspected Dirofilaria immitis infection, the other had a confirmed D. lutrae infection, and the red panda had a confirmed Dirofilaria sp. infection, presumably with D. immitis. Postmortem examinations revealed similar gross lesions, although they were less severe in the red panda. The trachea and primary bronchi contained abundant foamy fluid, the lungs were mottled with areas of consolidation, and the pulmonary parenchyma exuded abundant fluid at the cut section. Histologic evaluation revealed acute pulmonary edema, which resulted in respiratory failure and death. There may have been direct pulmonary cellular toxicity of melarsomine dihydrochloride or a severe systemic anaphylactic reaction to antigens released after parasite death. An idiosyncratic drug reaction or a low therapeutic index of melarsomine probably caused the death of the three individuals. Melarsomine dihydrochloride use should be avoided in North American river otters and red pandas.

Fish sedation, analgesia, anesthesia, and euthanasia: considerations, methods, and types of drugs.

Fish display robust neuroendocrine and physiologic stress responses to noxious stimuli. Many anesthetic, sedative, or analgesic drugs used in other vertebrates reduce stress in fish, decrease handling trauma, minimize movement and physiologic changes in response to nociceptive stimuli, and can be used for euthanasia. But extrapolating from limited published anesthetic and sedative data to all fish species is potentially harmful because of marked anatomic, physiologic, and behavioral variations; instead, a stepwise approach to anesthetizing or sedating unfamiliar species or using unproven drugs for familiar species is advisable. Additionally, knowledge of how water quality influences anesthesia or sedation helps limit complications. The most common method of drug administration is through immersion, a technique analogous to gaseous inhalant anesthesia in terrestrial animals, but the use of injectable anesthetic and sedative agents (primarily intramuscularly, but also intravenously) is increasing. Regardless of the route of administration, routine preprocedural preparation is appropriate, to stage both the animals and the supplies for induction, maintenance, and recovery. Anesthetic and sedation monitoring and resuscitation are similar to those for other vertebrates. Euthanasia is most commonly performed using an overdose of an immersion drug but injectable agents are also effective. Analgesia is an area in need of significant research as only a few studies exist and they provide some contrasting results. However, fish have mu and kappa opiate receptors throughout the brain, making it reasonable to expect some effect of at least opioid treatments in fish experiencing noxious stimuli.