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INTRACYTOPLASMIC INCLUSIONS IN CIRCULATING LEUKOCYTES FROM AN EASTERN BOX TURTLE (Terrapene carolina carolina) WITH IRIDOVIRAL INFECTION Matthew Allender, DVM, MS,1* Michael Fry, DVM, Dipl ACVP (Clinical Pathology),2 Armando R. Irizarry, DVM, PhD, Dipl ACVP,3 Linden Craig, DVM, PhD, Dipl ACVP,2 April Johnson, DVM,4 and Michael Jones, DVM, Dipl ABVP (Avian)5 1University of Illinois, College of Veterinary Medicine, 2001 S. Lincoln Ave. Urbana, IL 61802 USA; 2Department of Pathology, University of Tennessee, College of Veterinary Medicine, 2407 River Dr., Knoxville, TN 37996-4545 USA; 3Eli Lilly and Company, Lilly Research Laboratories, PO Box 708, Greenfield, IN 46140 USA; 4University of Florida, College of Veterinary Medicine, PO Box 100126, Gainesville, FL 32608 USA; 5Department of Small Animal Clinical Sciences, University of Tennessee, College of Veterinary Medicine, 2407 River Dr., Knoxville, TN 37996-4545 USA Abstract A free-ranging adult female eastern box turtle (Terrapene carolina carolina) was presented to the University of Tennessee in October 2003 because of suspected trauma and blindness. Physical examination revealed lethargy, clear ocular and nasal discharges, and white oral and laryngeal plaques. Intracytoplasmic inclusions within heterophils and large mononuclear leukocytes were observed on routine blood smear examination. Postmortem findings included necrosis of epithelial and parenchymal cells with intracytoplasmic inclusions. Ultrastructurally, the leukocyte inclusions consisted of variably electron-dense granular material and viral particles consistent with the Iridoviridae family of viruses. The virus shares 100% sequence identity to a 420 base pair sequence of frog virus 3 (FV3) (family Iridoviridae, genus Ranavirus) as determined by polymerase chain reaction and gene sequencing targeting a portion of the Ranavirus major capsid protein gene. 2006 PROCEEDINGS AMERICAN ASSOCIATION OF ZOO VETERINARIANS 1 INTESTINAL, PERICLOACAL, AND RENAL ADENOCARCINOMAS IN TWO MARINE TOADS (Bufo marinus) Douglas P. Whiteside, DVM, DVSc,1* Stephen A. Raverty, DVM, PhD, Dipl ACVP,2 Liza I. Dadone, VMD,1 Chantal Proulx, DVM,1 and Sandra R. Black, DVM, Dipl Path1 1Calgary Zoo, 1300 Zoo Road NE, Calgary, Alberta, T2E 7V6, Canada; 2Animal Health Centre, British Columbia Ministry of Agriculture and Food, 1767 Angus Campbell Road, Abbotsford, British Columbia, V3G 2M3, Canada Abstract Neoplasia has been reported worldwide in amphibians, although there are few reports of intestinal and renal neoplasms, with the exception of the northern leopard frog (Rana pipiens) in which Rana herpesvirus-1-induced Lucké renal adenocarcinomas have been well described.1-3 Within a 3-mo period, two marine toads (Bufo marinus) presented with prominent swellings in their pelvic areas. The first toad was diagnosed with a soft tissue mass within the pelvic inlet that was compressing the distal colon. Ultrasound revealed the mass to be well vascularized and intimately associated with the intestine. The mass was surgically resected, however the toad was euthanatized 2 days later due to a severe colonic prolapse. Histopathology of the mass revealed a moderately well differentiated transmural adenocarcinoma of the intestine, with no evidence of intranuclear inclusions. The second toad presented with a marked circumferentially swollen cloaca. Ultrasonography revealed a highly vascular soft tissue mass with scattered small cystic areas and intermingled wisps of hyperechogenic densities. Despite supportive therapy, the toad continued to lose weight and died approximately 2 wk post presentation. On necropsy, in addition to the pericloacal mass, there were hemorrhagic foci noted in the kidneys. Histologically, multicentric adenocarcinoma of the kidneys and pericloacal tissue was diagnosed. Amphophilic to eosinophilic intranuclear inclusions were noted within the neoplastic renal parenchyma. The temporal spacing of the two cases, coupled with the intranuclear inclusions within the renal adenocarcinoma in the second toad, was suggestive of a viral etiology. Further testing, including electron microscopy, is underway to further define the nature of these neoplasms. LITERATURE CITED 1. Davison, A.J., W. Sauerbier, A. Dolan, C. Addison, and R.G. McKinnell. 1999. Genomic studies of the Lucké tumor herpesvirus (RaHV-1). J. Cancer Res. Clin. Oncol. 125 (3-4):232-238. 2. Green, D.E., and J.C. Harshbarger. 2001. Spontaneous neoplasia in amphibia. In: Wright, K.M., and B.R. Whitaker (eds.). Amphibian Medicine and Captive Husbandry. Krieger Publishing Company. Florida. Pp. 335- 400. 3. McKinnell R.G., and D.L. Carlson.1997. Lucké renal adenocarcinoma, an anuran neoplasm: studies at the interface of pathology, virology, and differentiation competence. J. Cell Physiol. 173(2):115-8. 2 2006 PROCEEDINGS AMERICAN ASSOCIATION OF ZOO VETERINARIANS SKIN FLAP RESTORATION OF A TRAUMATIC WOUND IN A CUBAN IGUANA (Cyclura nubila nubila) Ray L. Ball, DVM and Clif Martel, BS, MBA* Busch Gardens Tampa Bay, 3605 Bougainvillea Drive, Tampa FL 33612 USA Abstract A 5-yr-old female Cuban iguana (Cyclura nubila nubila) was presented to the Busch Gardens Tampa Bay Zoo Hospital after receiving a degloving injury to the left forearm from a male of the same species. A full-thickness, 3-cm portion of skin was removed extending from the carpus proximally to just below the elbow. The iguana was anesthetized with isoflurane via mask (IsoSol®, VEDCO, Inc., St. Joseph, Missouri 64507 USA), then intubated for assessment and treatment. Radiographs confirmed no fractures of the area and no other injuries. Because of the fresh nature of the wound a skin graft approach was considered to heal this wound primarily. The arm and the left lateral thorax were prepared in routine fashion for surgery. A flap of skin 2 cm wide by 3 cm tall was created on the left lateral thorax just caudal to the axilla. This flap was deep enough that the panniculus muscle layer was included. The flap was created in a similar fashion as described for a surgical treatment of a hygroma of the elbow in dogs.1 The flap was created to encompass the entire wound. The arm was then extended and adducted to the body wall with the newly created flap wrapping around the defect. The surgical incision edges were sutured to the traumatic wound edges using simple interrupted sutures of 3-0 polydioxanone. A small roll of gauze was used to close the dead space where the limb met the body wall to lessen seroma formation. The limb was then wrapped in place to the body. Injectable enrofloxacin (Baytril 100®, Bayer Corp., Agriculture Division, Animal Health, Shawnee Mission, KS 66201 USA) was initiated at 5 mg/kg i.m. every 24 hr for 7 days. The bandage was changed periodically for 4 wk and the surgical site assessed, but little care was needed. At 4 wk post surgery, the site was surgically prepared and the flap was then excised from the body wall and wrapped around the limb. The flap was about 5 mm short of encompassing the entire circumference of the arm. This defect was managed with a biosynthetic absorbent wound dressing (BioDres®, DVM Pharmaceuticals, Inc., Miami, FL 33169 USA) and granulated well in the next 4 wk. The defect on the lateral thorax wall could not be entirely closed primarily and again was managed with the same bandaging materials. Four weeks after the second surgery the iguana had the bandages removed and it was released from the hospital. The surgical site has continued to look well over the past 2 yr. The skin of reptiles is similar to that of other animals in that it consists of an epidermis and a dermis. A major difference from mammals is that the epidermis has a keratin layer that can be thickened into scales. Information of the cutaneous circulation in reptiles is scarce and relates primarily to thermoregulation. Because of this, the surgeon must assume that the distribution of vessels is generally the same in quadrupeds. Ensuring that the panniculus layer is incorporated into the graft will ensure a reasonable blood supply to the graft. Immobilization of the graft was thought to be essential in the management of this wound and the location of the wound facilitated 2006 PROCEEDINGS AMERICAN ASSOCIATION OF ZOO VETERINARIANS 3 this. Avoiding dead space and seroma formation is also a must for a graft to take to a new location. The complication in this case was minimal and directly related to not accurately sizing the wound and allowing any margin of error in the initial graft. Careful planning is needed before any skin grafting is attempted. Traumatic injuries are not uncommon in zoological collections. If fresh, they are good candidates for primary healing with grafting techniques. Infected wounds must be aggressively treated first and a healthy granulation bed produced before any attempts can be made. Following basic principles of skin grafting will provide a reasonable guide to surgical repair of wounds in iguanids and perhaps other reptiles with similar skin types. LITERATURE CITED 1. Pope, E.R. 1990. Surgical treatment of hygroma of the elbow. In: Bojrab, M.J. (ed.) Current Techniques in Small Animal Surgery. Lea & Febiger, Philadelphia, PA. Pp. 496-489. 4 2006 PROCEEDINGS AMERICAN ASSOCIATION OF ZOO VETERINARIANS REFERENCE HEMATOLOGY AND SERUM CHEMISTRY VALUES FOR THE DIAMONDBACK TERRAPIN (Malaclemys terrapin) Ralph E. Werner, VMD Richard Stockton College of New Jersey, Jim Leeds Road, Pomona, NJ 08240 USA Abstract The diamondback terrapin (Malaclemys terrapin) is an inhabitant of the coastal marshlands of the eastern and southern United States. This unique turtle spends its life in tidewater creeks and ponds. Pressure from hunting, coastal construction, vehicle traffic, oil spills, and predation has prompted conservation efforts to protect the diamondback. Health assessment is a vital aspect of species management and conservation but can be quite difficult if reference blood values are not available. Prior to collecting blood samples, a small study was done to determine the optimal collection site in the diamondback.7 The femoral vein proved to be the best vessel for obtaining blood that was not diluted by lymph, so it was used throughout this study.