PULMONARY AND COELOMIC MYCOSES DUE TO METARHIZIUM AND BEAUVERIA SPECIES IN REPTILES.

This report documents cases of fatal pulmonary mycosis caused by entomopathogenic fungi in the genera Metarhizium and Beauveria (Order Hypocreales) in a loggerhead sea turtle (Caretta caretta), a Chinese alligator (Alligator sinensis), two gopher tortoises (Gopherus polyphemus), a Cuvier's dwarf caiman (Paleosuchus palpebrosus), a false gharial (Tomistoma schlegelii), a green sea turtle (Chelonia mydas), and a Kemp's ridley sea turtle (Lepidochelys kempii), and a case of granulomatous coelomitis in a hawksbill sea turtle (Eretmochelys imbricata). Fungi identified in these cases included Beauveria bassiana, Beauveria brongniartii, Metarhizium anisopliae, Metarhizium robertsii, and one case of infection by a novel Metarhizium species. The animals were either housed at zoos or brought into rehabilitation from the wild. Although the majority of animals had comorbidities, the fungal infections were believed to be the primary cause of death. Fungal susceptibility testing was performed on two Beauveria spp. isolates, and revealed lower minimum inhibitory concentrations for itraconazole and voriconazole when compared to terbinafine and fluconazole. This case series demonstrates that a variety of reptile species from different orders are vulnerable to infection with Metarhizium, and multiple species of sea turtle are susceptible to infection with Beauveria.

Endoscopic sexing in turtles and tortoises: 467 cases (2007-2017).

BACKGROUND: This multi-institutional retrospective study evaluated the feasibility and safety of endoscopic sex identification in 467 turtles and tortoises, representing 10 species. METHODS: Medical records of turtles and tortoises that underwent endoscopic sex identification at the University of Georgia, New England Aquarium and Turtle Conservancy were reviewed for presurgical management, anaesthesia, endoscopic equipment and surgical techniques, endoscopic results and complications. RESULTS: The majority of animals weighed less than 200 g, were fasted and anaesthetised using an injectable combination of ketamine, dexmedetomidine and morphine or hydromorphone, supplemented by local lidocaine at the prefemoral site. Anaesthetic reversal using atipamezole alone or in combination with naloxone was routine. For uncomplicated procedures, mean total anaesthesia, surgery and recovery times were 22, 4 and 18 minutes, respectively. All animals were placed in lateral recumbency for a prefemoral endoscopic approach to the coelom using a rigid telescope and sterile fluid infusion to visualise the gonads. Sex identification was definitive in 99.4% (n = 464) of the animals. Iatrogenic bladder perforation was the most common complication (n = 5), which necessitated extended anaesthesia and surgical time for repair. Only a single anaesthetic-related death was reported, which was associated with human error and drug overdose. CONCLUSIONS: This is the first large-scale study to retrospectively evaluate endoscopic sex identification in multiple chelonian species. Results suggest that endoscopic sexing is a safe, accurate and practical means for sex identification in turtles and tortoises, and represents a valuable tool in their reproductive management.

Osteomyelitis in cold-stunned Kemp's ridley sea turtles (Lepidochelys kempii) hospitalized for rehabilitation: 25 cases (2008-2018).

OBJECTIVE: To characterize osteolytic lesions in cold-stunned Kemp's ridley sea turtles (Lepidochelys kempii) hospitalized for rehabilitation and describe methods used for the management of such lesions. ANIMALS: 25 stranded, cold-stunned Kemp's ridley sea turtles hospitalized between 2008 and 2018. PROCEDURES: Medical records of sea turtles with a diagnosis of osteolytic lesions were reviewed retrospectively to obtain the date of diagnosis, clinical signs, radiographic findings, microbial culture results, hematologic and plasma biochemical data, cytologic and histologic findings, antimicrobial history, time to first negative culture result, treatment duration, and outcome. RESULTS: Lesions were identified radiographically a median of 50 days after admission and were located within epiphyses or metaphyses of various appendicular joints. Lesions were associated with periarticular swelling (n = 24), lameness (16), lethargy (2), and hyporexia (2). Bacterial culture yielded growth of single organisms (n = 16), multiple organisms (2), or no growth (6). Significant differences in hematologic and biochemical data were detected between the times of diagnosis and convalescence. Cytologic and histologic findings characterized the lesions as osteomyelitis leading to septic arthritis. Sixteen sea turtles were managed medically, and 8 were managed medically and surgically. Surgery resulted in rapid improvement in joint mobility and overall clinical status. Most (22/25 [88%]) sea turtles survived and were released after long-term management. CONCLUSIONS AND CLINICAL RELEVANCE: During rehabilitation, cold-stunned Kemp's ridley sea turtles may be affected by osteomyelitis. Medical management based on antimicrobial susceptibility testing was effective for most turtles. Long term management efforts in turtles are justified by high survival rate.

Fibropapillomatosis and Chelonid Alphaherpesvirus 5 Infection in Kemp's Ridley Sea Turtles (Lepidochelys kempii).

Fibropapillomatosis (FP), a debilitating, infectious neoplastic disease, is rarely reported in endangered Kemp's ridley sea turtles (Lepidochelys kempii). With this study, we describe FP and the associated chelonid alphaherpesvirus 5 (ChHV5) in Kemp's ridley turtles encountered in the United States during 2006-2020. Analysis of 22 case reports of Kemp's ridley turtles with FP revealed that while the disease was mild in most cases, 54.5% were adult turtles, a reproductively valuable age class whose survival is a priority for population recovery. Of 51 blood samples from tumor-free turtles and 12 tumor samples from turtles with FP, 7.8% and 91.7%, respectively, tested positive for ChHV5 DNA via quantitative polymerase chain reaction (qPCR). Viral genome shotgun sequencing and phylogenetic analysis of six tumor samples show that ChHV5 sequences in Kemp's ridley turtles encountered in the Gulf of Mexico and northwestern Atlantic cluster with ChHV5 sequences identified in green (Chelonia mydas) and loggerhead (Caretta caretta) sea turtles from Hawaii, the southwestern Atlantic Ocean, and the Caribbean. Results suggest an interspecific, spatiotemporal spread of FP among Kemp's ridley turtles in regions where the disease is enzootic. Although FP is currently uncommon in this species, it remains a health concern due to its uncertain pathogenesis and potential relationship with habitat degradation.

Venous blood gas and biochemical analysis of wild captured green turtles (Chelonia mydas) and Kemp's ridley turtles (Lepidochelys kempii) from the Gulf of Mexico.

Blood was collected from wild captured green and Kemp's ridley turtles off the west coast of Florida, USA. Blood gases and biochemical values were analyzed using a point of care (POC) device in the field. Analytes include pH, partial pressure of carbon dioxide (pCO2), partial pressure of oxygen (pO2), total carbon dioxide (TCO2), bicarbonate (HCO3), base excess (BE), oxygen saturation (sO2), lactate, sodium (Na), potassium (K), chloride (Cl), total carbon dioxide (TCO2), anion gap, ionized calcium, glucose, blood urea nitrogen (BUN), creatinine (Crea), hematocrit (Hct), and hemoglobin (Hb). These are novel data for wild healthy Kemp's ridley turtles, and results for green turtles were generally consistent with past studies of green turtles with exceptions primarily in blood gas values. Ninety percent of the green turtles had fibropapillomatosis (FP), but none of the blood analytes were correlated with disease severity. Only BUN was correlated with weight of green turtles, and there was no correlation between blood parameters and weight of Kemp's ridley turtles. This study provides data that are useful in understanding the physiologic status of sea turtles specific to this region, allowing for comparisons to other populations, life stages, and disease states.

TEMPORAL STABILITY OF IN VITRO VENOUS BLOOD GAS, pH, AND LACTATE VALUES OF COWNOSE RAYS (RHINOPTERA BONASUS) AND RED-EARED SLIDER TURTLES (PSEUDEMYS SCRIPTA ELEGANS).

This study assessed the in vitro temporal changes that occur in blood pH and lactate concentrations for an elasmobranch species and a chelonian species, as well as blood gases (partial pressures of carbon dioxide [pCO2] and oxygen [pO2]) for a chelonian species, with a portable clinical point-of-care analyzer. Blood samples were collected from 10 cownose rays (Rhinoptera bonasus) and 10 red-eared sliders (Pseudemys scripta elegans), stored on ice, and serially analyzed at six time points up to 90 min postcollection. Results indicate that analysis should be conducted as soon as possible after blood collection for these species, with immediate analysis being preferred. However, if analysis must be delayed, syringes may be capped, placed on ice, and analyzed at a later time. Analysis within 90 min provided clinically acceptable results for pH and lactate in both species and for pCO2 in red-eared sliders, whereas substantial artifactual increases of pO2 were seen in red-eared sliders.

Sulawesi tortoise adenovirus-1 in two impressed tortoises (Manouria impressa) and a Burmese star tortoise (Geochelone platynota).

Sulawesi tortoise adenovirus-1 (STAdV-1) is a newly discovered virus infecting endangered and threatened tortoises. It was initially described from a confiscated group of 105 Sulawesi tortoises (Indotestudo forsteni) obtained by the Turtle Survival Alliance and distributed to five sites with available veterinary care across the United States. In a 3-yr period from the initial outbreak, one multi-species collection that rehabilitated and housed adenovirus-infected Sulawesi tortoises experienced deaths in impressed tortoises (Manouria impressa) and a Burmese star tortoise (Geochelone platynota). Impressed tortoises that died had evidence of systemic viral infection with histopathologic features of adenovirus. Adenovirus was identified by consensus nested polymerase chain reaction (PCR) testing and subsequent sequencing of PCR products. Sequencing indicated that the adenovirus infecting these impressed tortoises and Burmese star tortoise was STAdV-1. In one impressed tortoise, viral infection was confirmed using transmission electron microscopy. In situ hybridization using a semiautomated protocol and fluorescein-labeled riboprobe identified STAdV-1 inclusions in spleen, liver, kidney, and testis of one impressed tortoise. The impact of this virus on captive and wild populations of tortoises is unknown; however, these findings indicate that STAdV-1 can be transmitted to and can infect other tortoise species, the impressed tortoise and Burmese star tortoise, when cohabitated with infected Sulawesi tortoises.

Myxobolus albi infection in cartilage of captive lumpfish (Cyclopterus lumpus).

Myxobolus albi was diagnosed in the cartilage of captive lumpfish (Cyclopterus lumpus) from 2 public aquaria. Eleven fish were affected, with the most common clinical signs being exophthalmos and grossly visible 1- to 2-mm white to tan scleral nodules. Myxozoan cysts were identified in the cartilage of the skull, branchial arch, sclera, vertebrae, tongue, all fin insertions, and the pectoral girdle. Cysts resulted in expansile, deforming, space-occupying lesions, resulting in exophthalmos but often lacking significant tissue damage or inflammation. Once cysts ruptured, free spores elicited a mild to marked inflammatory response. Spores measured 7.5 to 9.0 µm × 3.0 to 6.0 µm and contained 2 pyriform polar capsules oriented at one pole as well as occasional 1-µm-diameter basophilic nuclei. Identification was based on spore morphology together with polymerase chain reaction and sequence comparison of 18S ribosomal DNA. Isolates had 99% similarity to M. albi.

Hematologic values of captive Acadian redfish (Sebastes fasciatus).

The Acadian redfish (Sebastes fasciatus) is a popular display animal in public aquaria and an important fisheries resource, but hematologic values for this species have not been reported. This study investigated hematocrit, leucocyte counts, and leucocyte differential counts for 29 captive Acadian redfish collected in 2006 and 2008. All fish were judged to be in good health based on behavior, appetite, physical examination, skin cytology, and gill biopsy. Fish were anesthetized with tricaine methanesulfonate (MS-222) for blood collection from the ventral coccygeal vasculature. In 2008, MS-222 was buffered with sodium bicarbonate. Hematocrit values ranged from 27% to 45% (mean = 33% in 2006, mean = 31% in 2008). Leucocyte counts ranged from 670 to 35,000 cells/microl (mean = 6430 cells/microl in 2006, and 13,130 cells/microl in 2008). Lymphocytes were the most numerous of the leucocytes, followed by neutrophils and monocytes. Eosinophils and basophils were not seen. Fish weights were significantly greater in 2006, whereas total leucocyte counts, lymphocyte counts, and monocyte counts were significantly greater in 2008. Differences in leucogram values between the two groups may have been due to differences in size, age, anesthetic method, or other undetected factors. This is the first report of leucogram values for a species of the family Scorpaenidae. In general, values were similar to those of other teleosts, in which leucocyte counts can be quite variable.

Endoscopy and endosurgery of the chelonian reproductive tract.

The development of endosurgical techniques for chelonians has reduced the need for more invasive approaches such as plastron osteotomy. Surgical access and manipulation of much of the coelomic viscera of chelonians can be accomplished using endoscopy. Endoscopic methods may be used to perform many chelonian reproductive surgical procedures, including oophorectomy, salpingotomy, salpingectomy, gender identification, and removal of ectopic eggs.

Pathologic and parasitologic findings of cold-stunned Kemp's ridley sea turtles (Lepidochelys kempii) stranded on Cape Cod, Massachusetts, 2001-2006.

Necropsy reports for 28 stranded, cold-stunned Kemp's ridley sea turtles (Lepidochelys kempii) that died between 2001 and 2006 were reviewed retrospectively. Gross and microscopic lesions were compiled to describe the pathologic and parasitologic findings in turtles that were found freshly dead on the beach or that died within 48 hr of stranding. Anatomic lesions of varying severity were identified in each of the examined turtles and were identified in tissues of the alimentary, respiratory, integumentary, nervous and sensory, and urogenital systems in order of decreasing frequency. Necrotizing enterocolitis and bacterial or fungal pneumonia were the most frequently encountered lesions that were considered clinically significant. Parasites and parasitic lesions were identified primarily in tissues of the alimentary system and included intestinal cestodiasis and parasitic granulomas containing larval cestodes or nematodes. Postlarval cestodes were also found in the coelom of two turtles. In many cases, the extent and severity of lesions were judged to be insufficient to have solely caused mortality, suggesting that additional factors such as metabolic, respiratory, and electrolyte derangements; hypothermia; and drowning may be important proximate causes of death in cold-stunned turtles. Results of this study provide insight into pathologic conditions that may be of clinical relevance to rehabilitation efforts for cold-stunned sea turtles.

Systemic adenovirus infection in Sulawesi tortoises (Indotestudo forsteni) caused by a novel siadenovirus.

A novel siadenovirus was identified in the Sulawesi tortoise (Indotestudo forsteni). A group of 105 Sulawesi tortoises was obtained by the Turtle Survival Alliance. Many of the tortoises were in poor health. Clinical signs included anorexia, lethargy, mucosal ulcerations and palatine erosions of the oral cavity, nasal and ocular discharge, and diarrhea. Initial diagnostic tests included fecal testing for parasites, complete blood count and plasma biochemical analysis, mycoplasma serology, and polymerase chain reaction (PCR) testing for intranuclear coccidia and chelonian herpesvirus. Treatment included administration of antibiotics, antiparasitic medications, parenteral fluids, and nutritional support. Tissue samples from animals that died were submitted for histopathologic evaluation. Histopathologic examination revealed systemic inflammation and necrosis associated with intranuclear inclusions consistent with a systemic viral infection in 35 tortoises out of 50 examined. Fecal testing results and histopathologic findings revealed intestinal and hepatic amoebiasis and nematodiasis in 31 animals. Two of 5 tortoises tested by PCR were positive for Chlamydophila sp. Aeromonas hydrophila and Escherichia coli were cultured from multiple organs of 2 animals. The mycoplasma serology and PCR results for intranuclear coccidia and chelonian herpesvirus were negative. Polymerase chain reaction testing of tissues, plasma, and choanal/cloacal samples from 41 out of 42 tortoises tested were positive for an adenovirus, which was characterized by sequence analysis and molecular phylogenetic inference as a novel adenovirus of the genus Siadenovirus. The present report details the clinical and anatomic pathologic findings associated with systemic infection of Sulawesi tortoises by this novel Siadenovirus, which extends the known reptilian adenoviruses to the chelonians and extends the known genera of reptilian Adenoviridae beyond Atadenovirus to include the genus Siadenovirus.

Anesthesia, diagnostic imaging, and surgery of fish.

Anesthesia, diagnostic imaging, and surgery of fish have become routine parts of aquatic animal medicine. Anesthesia may be required for simple clinical procedures, diagnostic testing, or more involved surgery. Diagnostic modalities, including radiology, ultrasonography, and endoscopy, can be readily applied to fish and may provide valuable information. Despite some unique challenges, surgery can be performed in fish using basic surgical skills and principles and should be considered as a valid treatment option.

Disseminated phaeohyphomycosis in weedy seadragons (Phyllopteryx taeniolatus) and leafy seadragons (Phycodurus eques) caused by species of Exophiala, including a novel species.

During the period from January 2002 to March 2007, infections by melanized fungi were identified with greater frequency in aquarium-maintained leafy seadragons (Phycodurus eques) and weedy seadragons (Phyllopteryx taeniolatus), pivotal species to the educational and environmental concerns of the aquarium industry and conservation groups. The objective of this study was to characterize the pathology and identify fungi associated with phaeohyphomycotic lesions in these species. Samples from 14 weedy and 6 leafy seadragons were received from 2 institutions and included fresh, frozen, and formalin-fixed tissues from necropsy and biopsy specimens. Fresh and frozen tissues were cultured for fungi on Sabouraud dextrose agar only or both Sabouraud dextrose agar and inhibitory mold agar with gentamicin and chloramphenicol at 30 degrees C. Isolates were processed for morphologic identification and molecular sequence analysis of the internal transcribed spacer region and D1/D2 domains of the large subunit ribosomal RNA gene. Lesions were extensive and consisted of parenchymal and vascular necrosis with fungal invasion of gill (11/20), kidney (14/20), and other coelomic viscera with or without cutaneous ulceration (13/20). Exophiala sp. isolates were obtained from 4 weedy and 3 leafy seadragons and were identified to species level in 6 of 7 instances, namely Exophiala angulospora (1) and a novel species of Exophiala (5), based on nucleotide sequence comparisons and phylogenetic analyses. Disseminated phaeohyphomycosis represents an important pathologic condition of both weedy and leafy seadragons for which 2 species of Exophiala, 1 a novel species, have been isolated.

Piscine patients: basic diagnostics.

This article can serve as an introduction for practitioners who are interested in adding fish to their practice. It reviews the current literature, includes contributions from institutional collaborators, and draws on clinical expertise from diverse cases and applied research from the Animal Health Department at the New England Aquarium. Basic diagnostic approaches to piscine patients are featured. We hope to inspire other veterinary professionals to pursue diagnostics, perform surgery, and practice preventive medicine in aquatic animal patients.

Coelioscopic-assisted prefemoral oophorectomy in chelonians.

OBJECTIVE: To describe a coelioscopic-assisted prefemoral oophorectomy technique for use in chelonians. DESIGN: Descriptive report. ANIMALS: 11 adult female turtles (6 red-eared sliders, 2 box turtles, 1 painted turtle, 1 four-eyed turtle, and 1 Chinese red-necked pond turtle). Five turtles required oophorectomy because of reproductive tract disease; the remaining 6 underwent elective oophorectomy. PROCEDURES: Turtles were anesthetized and positioned in dorsal recumbency. An incision was made in the prefemoral fossa, and a 2.7-mm rigid endoscope was inserted into the coelomic cavity and used to identify the ovaries. Each ovary was grasped with forceps and exteriorized through the prefemoral incision. The ovarian vasculature was ligated, and the mesovarium was transected. Closure was routine. RESULTS: In 8 turtles, bilateral oophorectomy was performed through a single incision. In 2 turtles, unilateral oophorectomy was performed in an attempt to maintain reproductive potential. In 1 turtle with a unilateral ovarian remnant from a previous surgery, unilateral oophorectomy was performed. Nine turtles recovered. One box turtle with severe hepatic lipidosis died 7 days after surgery. A second box turtle died 2 days after removal of retained eggs and a large bacterial granuloma. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that coelioscopic-assisted prefemoral oophorectomy is a practical and safe method for treating reproductive disorders and performing elective oophorectomy in turtles. This technique represents a potential alternative to plastron osteotomy in sexually mature chelonians.

Chelonian reproductive disorders.

The study of reproductive diseases of chelonians has become increasingly sophisticated in the last decade. Widespread captive breeding has increased the number of reproductive problems presented to veterinarians. Advances in the level of veterinary care have encouraged chelonians owners to seek solutions to problems that may have previously been considered unsolvable. Improved diagnostic imaging, particularly radiography, ultrasonography and endoscopy, have made visualization and diagnosis of reproductive disease easier. Better quality veterinary care has made treatment of chelonian diseases more sophisticated which has lead to improved methods of anesthesia, surgery, and medicine. Concerns over rapidly diminishing chelonian populations and habitat have made the need for study of reproduction a higher priority; veterinary medicine has gained from this research.