The role of Virus "X" (Tortoise Picornavirus) in kidney disease and shell weakness syndrome in European tortoise species determined by experimental infection.

Tortoise Picornavirus (ToPV) commonly known as Virus "X" was recently discovered in juvenile European tortoises suffering from soft carapace and plastron as well as kidney disease. Therefore, this virus was a potential candidate to be a causative agent for these disease patterns. Spur thighed tortoises (Testudo graeca) seemed to be more susceptible to establish clinical symptoms than other European species like T. hermanni. Thus this trial investigated the role of ToPV in the described syndrome. Two groups of juvenile European tortoises (T. graeca and T.hermanni) each of 10 animals, were cloacally, oronasally and intracoelomically inoculated with an infectious dose (~ 2000 TICD) of a ToPV strain isolated from a diseased T. graeca. A control group of two animals of each species received non-infected cell culture supernatant. The tortoises were examined daily and pharyngeal and cloacal swabs for detection of ToPV-RNA by RT-PCR were taken from each animal every six days for a period of 6 months. At the end of the study the remaining animals were euthanised and dissected. Bacteriological and parasitological tests were performed and organ samples of all tortoises were investigated by RT-PCR for the presence of ToPV and histopathology. Animals that were euthanised at the end of the experiment, were examined for presence of specific anti-ToPV antibodies. Several animals in both inoculated groups showed retarded growth and a light shell weakness, in comparison to the control animals. Three animals were euthanised during the trial, showing reduced weight gain, retarded growth, severe shell weakness and apathy, in parallel to clinical observations in naturally infected animals. In all inoculated animals of both species an intermittent virus shedding, starting from 18 days post inoculation (d.p.i.), till 164 d.p.i. was detected, while the control animals remained negative. The virus was successfully reisolated in terrapene heart cell culture in 16 of 20 inoculated animals of both species. Histopathology of most inoculated animals revealed a lack of bone remodeling and vacuolisation in kidney tubuli which supports the described pathogenesis of nephropathy and osteodystrophy. Anti- ToPV antibody titres ranged from 1:2 to >1:256 in 13 of 20 animals, whereas all control animals were seronegative. The study proofed the Henle Koch`s postulates of ToPV as causative agent for shell dystrophy and kidney disease in both testudo species. The proposed species specific sensitivity towards clinical disease was not observed.

Endoscopic assisted orchiectomy in Herman's tortoises (Testudo hermanni sp.).

OBJECTIVE: Male tortoises in captivity are often aggressive against other males or females, in particular during mating season related to hormonal influences (testosterone). Castration in males is the treatment of choice in many vertebrate species. A novel technique of minimal invasive castration is presented for Herman's tortoises (Testudo hermanni). MATERIAL AND METHODS: The procedure was performed in ten mature males. An endoscope (2.7 mm/30° angled) was inserted into a prefemoral incision on each side. The testicle was resected after ligation of the gubernaculum testis and the spermatic cord using hemoclips. RESULTS: Each testicle could be removed in approximately 20 minutes. Complications like hemorrhage or damage of adjacent tissue did not occur and all animals recovered uneventfully. CONCLUSIONS AND CLINICAL RELEVANCE: Using the bilateral prefemoral entrance offers an alternative option for orchiectomy in Herman's tortoises without dissection of the shell. Thus complications like hemorrhage or impaired wound healing followed by sequestration of the bone flap are prevented. Because of the anatomical settings and the risk of tissue damage and time consumption, a unilateral approach is not recommended. Fasting the animals is necessary due to the voluminous gastrointestinal tract of this herbivorous tortoise and emptying the urinary bladder provides more space for manipulations in the coelomic cavity and prevents clipping and cutting of adjacent organs. The magnification via the endoscope is beneficial for orientation in the coelomic cavity, which is formed by the extremely convex carapace. Without this equipment it is challenging to visualize the dorsocaudally located gonads through the relatively small incision and the inserted instruments might block the view at the surgical field. Administration of hemoclips achieved a good hemostasis and the testicles could be resected without major blood loss. The described technique is a gentle method for resection of the testicles in this species and can be adapted to other European tortoise species of equal size.