Herpesviruses are well-known infectious agents with remarkably wide host ranges. Starting in 1975 (33), several reports have documented the presence of herpesvirus-like particles in land tortoises and freshwater and marine turtles (5, 7-9, 11, 15, 17, 19-23, 27-29, 30, 38). Recent investigations have revealed an association between the presence of herpesvirus and an upper respiratory tract disease in Mediterranean tortoises [spur-thighed tortoise (Testudo graeca) and Hermann's tortoise (T. hermanni)] (5,8,9,17,20,23,(27)(28)(29)30).In tortoises with herpesvirus infection, clinical signs range from a mild conjunctivitis to a severe stomatitis-glossitis and pharyngitis. Diphtheritic plaques can be observed on the dorsal surface of the tongue and on the hard palate of infected tortoises. Frequently, a clear serous to a mucopurulent nasal discharge is present. Signs of central nervous system disease have also been reported in Mediterranean tortoises with herpesvirus infection (17).Eosinophilic intranuclear inclusions, often seen in multiple tissues, are particularly prominent in tortoises with pharyngitis and glossitis. As seen with transmission electron microscopy, inclusions consist of numerous viral particles. The morphology and morphogenesis have been used to categorize the virus as herpesvirus.A diagnosis of herpesvirus infection is often made based solely upon light or electron microscopy findings. Antemortem diagnosis can be made using biopsy specimens of oral lesions. A serum neutralization (SN) test has been developed but is limited in its application since it is only available in a few research laboratories in Europe (10). In addition, time is a limiting factor with the SN test. Ten to 14 days are required to obtain the final reading and a laborious procedure is required. An easier and faster but equally reliable serodiagnostic test is needed. In this report, we describe the development of an enzyme-linked immunosorbent assay (ELISA) that can be used to monitor the exposure to herpesvirus of free-range, private, and zoo collections of tortoises. MATERIALS AND METHODSViruses. Herpesvirus isolates HV1976 and HV4295/7R/95 were used as antigens in the ELISAs and immunoblotting. HV1976 was isolated from a captive Hermann's tortoise from the United States (Washington), while HV4295/7R/95 was isolated from a captive Hermann's tortoise in Germany during a herpesvirus outbreak in a private collection (27).Antigen preparation for ELISA. The herpesvirus isolates were grown in terrapene heart cell monolayers (TH-1; ATCC-CCL 50 Sub-line B1; American Type Culture Collection, Rockville, Md.) in T-150 plastic flasks with ventilated caps (Corning, Rochester, N.Y.) for use as ELISA antigens. The TH-1 cells were grown in Dulbecco's modified Eagle's medium F12 (Gibco BRL, Grand Island, N.Y.) with 5% fetal bovine serum (Sigma, St. Louis, Mo.), gentamicin (60 mg/liter) (Sigma), penicillin G (120,000 U/liter), streptomycin (120,000 U/liter), and amphotericin B (300 g/liter) (ABAM; Sigma). Infected cell monolayers were scraped a...
Ranaviral disease in amphibians has been studied intensely during the last decade, as associated mass-mortality events are considered to be a global threat to wild animal populations. Several studies have also included other susceptible ectothermic vertebrates (fish and reptiles), but only very few cases of ranavirus infections in lizards have been previously detected. In this study, we focused on clinically suspicious lizards and tested these animals for the presence of ranaviruses. Virological screening of samples from lizards with increased mortality and skin lesions over a course of four years led to the detection of ranaviral infections in seven different groups. Affected species were: brown anoles (Anolis sagrei), Asian glass lizards (Dopasia gracilis), green anoles (Anolis carolinensis), green iguanas (Iguana iguana), and a central bearded dragon (Pogona vitticeps). Purulent to ulcerative-necrotizing dermatitis and hyperkeratosis were diagnosed in pathological examinations. All animals tested positive for the presence of ranavirus by PCR and a part of the major capsid protein (MCP) gene of each virus was sequenced. Three different ranaviruses were isolated in cell culture. The analyzed portions of the MCP gene from each of the five different viruses detected were distinct from one another and were 98.4-100% identical to the corresponding portion of the frog virus 3 (FV3) genome. This is the first description of ranavirus infections in these five lizard species. The similarity in the pathological lesions observed in these different cases indicates that ranaviral infection may be an important differential diagnosis for skin lesions in lizards.
The metabolism of tortoises is influenced by many factors, which in turn also influences the clinical chemistry parameters in these animals. There are currently limited data available on species, sex, and season-specific reference intervals. The goal of this study was to establish new reference intervals for adult Hermann's tortoises (Testudo hermanni), which include variations according to season and sex. Lithium heparinized blood samples from 256 Hermann's tortoises were collected and biochemistry parameters (alkaline phosphatase [ALP], glutamate-dehydrogenase [GLDH], alanine-aminotransferase [ALT], aspartate-aminotransferase [AST], bile acids [BA], creatine kinase [CK], total protein [TP], albumin [Alb], urea, uric acid [UA], inorganic phosphorus [P], total calcium [Ca], sodium [Na], and potassium [K]) were measured from May 2016 to October 2017. The results showed many variations depending on sex and season. Male Hermann's tortoises had higher ALT, BA, CK, and UA concentrations, and females had higher Ca concentrations. The concentrations of BA, UA, GLDH, TP, and Alb decreased during the course of the year, and ALT increased over time in males. The present study demonstrates that it is important to establish specific reference intervals for each species that include seasonal and sex specific variations in order to facilitate correct interpretation of blood results.
The aim of this study was to describe the specific gross and radiographic anatomy of the digestive tract of inland bearded dragons ( Pogona vitticeps ). Eleven bearded dragon cadavers of both sexes (6 females, 5 males) were dissected to examine, measure, and document the specific gross anatomy of the alimentary canal. Measurements collected from the cadavers included snout-vent length, total length of the alimentary canal, and the lengths of the individual sections of the gastrointestinal tract, including the esophagus, stomach, small intestine, ampulla coli, isthmus coli, rectum, and the distance from the coprodeum to the vent opening. Twenty-two healthy adult bearded dragons (13 females, 9 males) maintained under standardized husbandry conditions underwent a physical examination, blood collection, and whole-body dorsoventral and lateral survey radiographs; these animals were used to provide the radiographic images of the complete digestive tract. For the subsequent contrast passage studies, two different contrast media, barium sulfate (BaSO 4 , Barilux suspension) and an iodinated ionic radiocontrast agent (Sodium meglumine amidotrizoate [SMAT], Gastrografin), were used. Water-diluted Barilux suspension (dose 9 ml/kg) was administered orally to 5 bearded dragons, while Gastrografin (dose 5ml/kg) was administered orally to 21 bearded dragons. Four animals were used for both contrast media studies, but received a break of four weeks in between. Dorsoventral and laterolateral radiographs were collected at 0 (baseline), 15, 30, and 45 minutes and 1, 2, 3, 4, 5, 6, 8, 10, 12, 24, 30, and 36 hours after each contrast medium was administered. Both contrast media were found to illustrate the alimentary tracts in the adult bearded dragons. Transit time was substantially faster with SMAT, and SMAT illustrated the entire gastrointestinal tract within 36 hours; BaSO 4 did not fully illustrate the gastrointestinal tract in 36 hours. These results might serve as a guideline for the interpretation of subsequent contrast studies in this lizard species.
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