Assessment of the effects of intramuscular administration of alfaxalone with and without medetomidine in Horsfield's tortoises (Agrionemys horsfieldii)

ObjectiveTo characterise four different intramuscular (IM) anaesthetic protocols, two with alfaxalone and two with alfaxalone in combination with medetomidine in terrestrial tortoises.Study designBlinded, randomized, cross‐over experimental study.AnimalsNine healthy adult male Horsfield's tortoises (Agrionemys horsfieldii).MethodsEach tortoise was randomly assigned to one of four different protocols: 1) 10 mg kg^?1 alfaxalone; 2) 10 mg kg^?1 alfaxalone + 0.10 mg kg^?1 medetomidine; 3) 20 mg kg^?1 alfaxalone; and 4) 20 mg kg^?1 alfaxalone + 0.05 mg kg^?1 medetomidine. During the experiment, the following variables were recorded: heart rate; respiratory rate; peripheral nociceptive responses; muscle strength; ability to intubate; palpebral, corneal and tap reflexes; and cloacal temperature.ResultsProtocols 1 and 2 resulted in moderate sedation with no analgesia, and moderate to deep sedation with minimal analgesia, respectively. Protocols 3 and 4 resulted in deep sedation or anaesthesia with variable analgesic effect; these two protocols had the longest total anaesthetic time and allowed intubation in 6/9 and 8/9 tortoises respectively. The total anaesthesia/sedation time produced by alfaxalone was significantly increased (p <0.05) by the addition of medetomidine. There were no significant differences regarding time to plateau phase and duration of plateau phase. Baseline heart rate of 53 ± 6 beats minute^?1 decreased significantly (p <0.05) with all protocols, and was lower (p <0.05) in protocols 3 and 4. Heart rate increased after atipamezole administration, but the increase was transient. In two tortoises, extreme bradycardia with no cardiac activity for 10 minutes was observed with protocols 3 and 4.Conclusion and clinical relevanceAlfaxalone 10 and 20 mg kg^?1 IM can be used for sedation for non‐painful procedures. Alfaxalone in combination with medetomidine can be used for deeper sedation or anaesthesia, but the observed respiratory and cardiovascular depression may limit its use.     

Russian tortoises (Agrionemys horsfieldi) as a potential reservoir for Salmonella spp

A total of 80 Russian tortoises brought in Poland were examined for presence of Salmonella. Salmonella was detected in 15 out of all the animals tested (18.75%). Of the total of 56 strains, 30 (53.57%) belonged to Salmonella enterica subsp. enterica (I) and 26 to Salmonella enterica subsp. salamae (II). The predominant serotype within subspecies I was S. Newport, which is one of the most serotypes causing salmonellosis in humans and warm-blooded animals. In vitro determination of the susceptibility of Salmonella to the 10 medicinal preparations showed that all tested strains were sensitive to norfloxacin, sulfamethoxazole with trimethoprim, florfenicol, gentamicin, tetracycline and ampicillin, resistance was noted only to amoxicillin with clavulanic acid (12 strains), and intermediate sensitivity to colistin (7 strains), enrofloxacin (2 strains) and cephalexin (5 strains). These studies confirmed that Russian tortoises are a significant reservoir for Salmonella and may represent a potential source of infection for humans.     

Herpesvirus infection in tortoises (Malacochersus tornieri and Testudo horsfieldii)

Large numbers of pancake tortoises (Malacochersus tornieri) and Horsfield tortoises (Testudo horsfieldii) in three consignments imported into Japan died soon after arrival. Some tortoises in the first consignment were dead on arrival. Postmortem examination of two of the pancake tortoises and four of the Horsfield tortoises revealed necrotizing lesions of the oral mucosa in both species, primarily in the tongue. Eosinophilic to amphophilic inclusion bodies were visible in the nuclei of mucosal epithelial cells in the lesions. Similar inclusion bodies were observed in the liver, spleen, adrenal glands, stomach, lungs, kidneys, small and large intestines, pancreas, and cerebrum of the pancake tortoises and in the liver, spleen, and pancreas of the Horsfield tortoises. Electron microscopic examination of the cells containing inclusion bodies showed herpesvirus-like particles about 100 nm in diameter in the cytoplasm. Nested polymerase chain reaction analysis using a herpesvirus consensus primer method confirmed the presence of a characteristic herpesvirus base sequence in tissue from these lesions.     

Chronic eosinophilic dermatitis associated with persistent feline herpes virus infection in cheetahs (Acinonyx jubatus)

A chronic ulcerative and eosinophilic dermatitis occurred in 20 captive cheetahs (Acinonyx jubatus) with persistent feline herpes virus 1 (FHV1) infection. Affected animals had erythematous, ulcerated plaques primarily on the face and forelegs in sites of contact with lachrymal and salivary secretions. The dermatitis was characterized by dense infiltrates of eosinophils and plasma cells and pseudoepitheliomatous hyperplasia. Rare keratinocytes within the lesions had nuclei with marginated chromatin and small eosinophilic inclusions composed of herpes virus nucleocapsids. Virus isolated from lesions was confirmed to be FHV1. Lesions persisted and progressed unless removed by cryoexcision. The occurrence of this unusual reaction to FHV1 in approximately 5% of captive North American cheetahs suggests a species propensity for a Th2-dominant response to herpes virus infection. This atypical immune reaction may indicate a heritable trait or modulation of the immune response by other factors such as chronic stress.     

Bovine leukemia virus infection in a juvenile alpaca with multicentric lymphoma

A 13-month-old alpaca (Vicugna pacos) was presented for mandibular masses and weight loss. Histopathology of biopsy tissue was consistent with lymphoma. The alpaca was euthanized and necropsy revealed lymphoma masses in multiple organs. Immunohistochemistry for T- and B-cell typing was inconclusive. Serology and in-situ polymerase chain reaction hybridization were positive for bovine leukemia virus.     

Ultrasonographic diagnosis of pericardial effusion and atrial dilation in a spur-thighed tortoise (Testudo graeca)

Ultrasonography was used to diagnose pericardial effusion, atrial dilatation and liver masses in a spur-thighed tortoise which was more than 80 years old and suffering from posthibernation anorexia, lethargy, oedema and pneumonia. The tortoise was treated twice with frusemide and ceftazidime for the pneumonia, resulting each time in a temporary remission for about a month. After a further recurrence, the animal was euthanased and the lesions predicted by ultrasound were confirmed postmortem. It is suggested that ultrasound may be useful for the differentiation of cardiac problems from other causes of posthibernation lethargy in the tortoise.     

Immunohistochemical diagnosis of persistent infection with Bovine Viral Diarrhea Virus (BVDV) on skin biopsies

Detection of persistent infection with BovineViral Diarrhea Virus (BVDV) is essential for both epidemiological and clinical reasons. In addition to the classical virological methods such as virus isolation in tissue culture, ELISA and RT-PCR, immunohistochemistry of skin biopsies has become a useful and reliable tool. Assuming that the presence of BVDV antigen in skin structures is restricted to persistent infection, this method could differentiate from transient infection. In order to answer this question, 6 calves were experimentally infected orally with a non-cytopathic genotype 1 BVDV strain belonging to the subtype k.The calves developed fever, mucopurulent nasal discharge, coughing and leucopenia with relative lymphopenia. Immunohistochemistry of skin biopsies taken daily up to day 13-post infection did not reveal any evidence of BVDV infection. BVDV was, however, isolated from blood samples on cell cultures. Anti-NS3-antibody-ELISA and serum neutralization tests showed that all six calves seroconverted. We conclude that in acute BVDV infections, with genotype 1 and the subtypes found in Switzerland (b, e, h and k) viral antigen is not found in epidermal structures of the skin. In contrast, persistently infected animals test positive for BVD viral antigen by immunohistochemistry of the skin.     

First evidence of Kunjin strain of West Nile virus associated with saltwater crocodile (Crocodylus porosus) skin lesions

Recently, the Kunjin strain of West Nile virus (WNV_KUN) has been detected using qRT‐PCR in belly skin lesions of farmed juvenile saltwater crocodiles. This follows an established association between similar lesions and West Nile virus in American alligators. The lesions present as cutaneous lymphohistiocytic aggregates in the dermal layers of both species. While these lesion do not create an obvious defect on the live crocodile, upon tanning the lesion area collapses and does not uptake the dye evenly, thus reducing its aesthetic appeal. As a result, skins are being rejected jeopardising the economic viability of the Australian crocodile industry. Over 50 skin lesions have since been confirmed as WNV_KUN‐positive and preliminary evidence of lesion restructuring is presented. Horizontal transmission of WNV_KUN by mosquitoes is well‐established but other transmission routes, such as ingestion and cloacal shedding, need further evaluation. An infection trial is currently underway to ensure WNV_KUN is the causative agent of these skin lesions.     

Endotheliotropic elephant herpes virus (EEHV) infection. The first PCR-confirmed fatal case in Asia

Since 1995, 4 suspected cases of Endotheliotropic Elephant Herpes Virus (EEHV) infection, i.e. based on clinical presentation, have occurred in Asia without resulting in epidemic outbreaks as expected. In order to confirm the presence of EEHV on the continent of Asia, viral DNA particles from liver samples of a wild-caught 3-year-old elephant found dead at a Cambodian elephant sanctuary and clinically diagnosed with EEHV, were PCR processed using known EEHV strain primers. The presence of EEHV viral nucleic acids was confirmed and the nucleic acids had a 99% sequence similarity to the U.S.A strain (gene bank locus: AF117265) and 97% sequence similarity to the European strain (gene bank locus: AF354746) assigning this case to the EEHV-1 cluster. More than the confirmation of EEHV on the continent of Asia, is the phylogenic relationship to the USA and European strains with no corresponding contact or transport of USA or European elephants to Asia. Thus, this brings many of the traditional theories into question. Although almost forgotten, this disease is still ramped in captive elephant populations worldwide and continues to devastate particularly the neonatal and weaning-age population. Special attention and continued research are needed specifically in the area of basic virology and epidemiology.