Symmetrical Dimethylarginine as a Diagnostic Parameter in Hermann's Tortoises (Testudo hermanni).

Background: Despite improvements in habitational conditions, kidney disease is relatively common in tortoises. Objectives: Purpose of this study was the establishment of Symmetrical dimethylarginine (SDMA) reference values for clinically healthy Hermann's Tortoises. Animals: Clinically healthy Hermann's Tortoises (n = 131) were included in the period from October 2017 to September 2019. Methods: Creatinine and other biomarkers were tested at IDEXX Laboratories, Germany using residual blood samples from Hermann's tortoises. SDMA was measured with the IDEXX test and verified by liquid chromatography-mass spectrometry at IDEXX Laboratories, USA. Results: SDMA values ranged from 1 to 21 µg/dl (n = 131) for the IDEXX SDMA Test and SDMA values ranged from 1 to 17 µg/dl (n = 82) for LC-MS. For the comparison of the two measuring systems, the following results were obtained R 2 = 0.75 (p < 0.001). Conclusion and Clinical Importance: SDMA can be measured in Hermann's Tortoises and the reference values range in clinically healthy animals is comparable to that of dogs and cats.

PLASMA CAPILLARY ZONE ELECTROPHORESIS AND PLASMA CHEMISTRY ANALYTES IN TORTOISES (TESTUDO HERMANNI, TESTUDO GRAECA) AND TURTLES (TRACHEMYS SCRIPTA ELEGANS, GRAPTEMYS SPP.) IN FALL.

Clinical health checks including blood testing before brumation in fall are an important tool in reptile medicine and help to reduce the risk of complications during brumation. Electrophoresis may be useful for the evaluation of liver cell function and the detection of antigenic stimulation. The goal of this study was to compare species-specific reference intervals for plasma chemistry analytes and capillary zone electrophoresis (CZE) for a variety of pet chelonian species in fall. Lithium heparinized samples were collected from 68 clinically healthy spur-thighed tortoises (Testudo graeca), 79 red-eared sliders (Trachemys scripta elegans), and 30 map turtles (Graptemys spp.) from September to November 2016 and 2017. During the same period, 128 equivalent samples were collected from Hermann's tortoises (Testudo hermanni) and the obtained data was used for comparison. Chemistry analytes were measured from plasma using an autoanalyzer and electrophoresis was carried out by CZE. Significant variations (P < 0.0001) between the species were found for several analytes including total protein, percent albumin, and albumin: globulin ratio, α-, ß-, and γ-globulin, alkaline phosphatase, glutamate dehydrogenase, alanine aminotransferase, aspartate aminotransferase, bile acids, creatine kinase, urea, uric acid, inorganic phosphorus, total calcium, and potassium. The variations in albumin (g/l) measured by CZE were also significant (P = 0.0064). No significant variations were detected for sodium levels. The results demonstrate the importance of species-specific reference intervals and provide reference intervals for the plasma chemistry and capillary zone electrophoresis in three chelonian species.

Body condition scoring (BCS) in corn snakes (Pantherophis guttatus) and comparison to pre-existing body condition index (BCI) for snakes.

In the veterinary profession, the body condition score (BCS) plays an important role in the assessment of patients. It is a subjective, tactile method of evaluating body fat and muscle mass and is used in numerous species. Recognizing obesity (or the contrary, emaciation) is important for veterinarians treating reptiles and could be facilitated by a BCS. An existing form of body condition assessment already used is the body condition index (BCI), where the residuals from a regression of body mass on body length are calculated. Therefore, the goal of this study was to provide practitioners with a BCS system for corn snakes (Pantherophis guttatus) and to test it against the BCI. A total of 22 corn snakes (Pantherophis guttatus), stationed at the "Auffangstation für Reptilien" in Munich (reptile rescue centre, RRC), were subject of this study. Each had the following measurements taken: body weight (BW), snout-tail tip length (STL), snout-vent length (SVL) and circumference in the middle (C). Manual palpation of spine, area between vertebral spinous and transverse process, ribs and neck of each snake was performed by three veterinarians and assigned to specific scores by each examiner. A BCS (mean of examiners' scores) was given to each snake according to manual palpation. The BCS system was chosen to be out of 5 in 0.5-point steps with 2.5 considered as ideal BCS. In the studied snakes, the BCS ranged from 1.5 to 3.5, with a median of 2.5. The median BW was 309 g (75-967 g), the median STL was 123 cm (79-153 cm), the median SVL was 104 cm (73-133 cm) and the median C was 7.5 cm (4.3-11 cm). BCS and BCI were positively correlated. A BCS includes a manual palpation of the animal and thus gives the examiner additional information to the objectively measured/calculated index.

A comparison of thyroid hormone levels and plasma capillary zone electrophoresis in red-eared sliders (Trachemys scripta elegans) and map turtles (Graptemys spp.) depending on season and sex.

BACKGROUND: Thyroid hormones and capillary zone electrophoresis are rarely studied in reptile medicine. OBJECTIVES: The goal of this prospective study was to establish RIs for plasma proteins and thyroid hormones, total tetraiodothyronine (tT4), free T4 (fT4), total triiodothyronine (tT3), and free T3 (fT3), as well as iodine blood levels in red-eared sliders (Trachemys scripta elegans) and map turtles (Graptemys spp.). METHODS: Variables were measured over 1 ½ years to determine variations depending on sex and season, as well as any correlations between the parameters. A total of 131 blood samples from red-eared sliders and 51 blood samples from map turtles were collected from May 2016 to November 2017. The levels of tT4, fT4, and fT3 were measured using standardized autoanalyzer test systems; tT3 was measured by ELISA and iodine inductively coupled plasma mass spectrometry. Total protein was quantitated using the biuret method, and plasma protein fractions were measured using capillary zone electrophoresis. RESULTS: The results showed significant variations in thyroid hormone levels and plasma protein fractions depending on sex and season. Total T4 and fT4 concentrations were significantly (P < .05) correlated in both turtle species and sexes in all seasons. Thyroid hormone levels correlated with blood proteins in a few seasons, including a positive correlation between tT4 and fT4 in female red-eared sliders in fall and female map turtles in summer and fall. CONCLUSIONS: This study demonstrates the importance of considering species, season, and sex variations when interpreting thyroid hormone and plasma protein levels. It also shows that thyroid hormone levels are not always influenced by total protein and albumin.

REFERENCE INTERVALS FOR PLASMA CAPILLARY ZONE ELECTROPHORESIS IN HERMANN'S TORTOISES (TESTUDO HERMANNI) DEPENDING ON SEASON AND SEX.

Electrophoresis is a useful diagnostic tool for the detection of shifts in plasma protein fractions, which can be associated with a variety of diseases. This method is not often used in reptile medicine because there are only a few studies available and no reference intervals have been published for most species. The goal of this study was to establish reference intervals for plasma capillary zone electrophoresis in Hermann's tortoises (Testudo hermanni) under consideration of variations by season and sex. 251 samples from healthy adult Hermann's tortoises were collected from May 2016 to October 2017. Total protein was measured by the biuret method and albumin was measured twice by bromocresol green dye-binding method (BCG) and capillary zone electrophoresis (CZE) in order to compare the detection methods. The globulin fractions were analyzed by CZE. Seasonal variations were significant for total protein (P = 0.0055), albumin (P = 0.0322) and α-globulins (P = 0.0007). The variations between male and female Hermann's tortoises were highly significant for albumin (P < 0.0001), α-globulins (P < 0.0001) and ß-globulins (P < 0.0001). The differences between the measurements of albumin by CZE and BCG also differed depending on the sex and season. This study demonstrates that it is important to include the variations by sex and season when establishing reference intervals and interpreting results. Furthermore, it confirms the importance of using electrophoresis for the correct determination of albumin in tortoises.

Prevalence of neutralising antibodies against adenoviruses in lizards and snakes.

Adenoviruses (AdVs) are relatively common in lizards and snakes, and several genetically distinct AdVs have been isolated in cell culture. The aims of this study were to examine serological relationships among lizard and snake AdVs and to determine the frequency of AdV infections in these species. Isolates from a boa constrictor (Boa constrictor), a corn snake (Pantherophis gutattus) and a central bearded dragon (Pogona vitticeps), and two isolates from helodermatid lizards (Heloderma horridum and H. suspectum) were used in neutralisation tests for the detection of antibodies in plasma from 263 lizards from seven families (including 12 species) and from 141 snakes from four families (including 28 species) from the USA and Europe. Most lizard and snake samples had antibodies against a range of AdV isolates, indicating that AdV infection is common among these squamates. Neutralisation tests with polyclonal antibodies raised in rabbits demonstrated serological cross-reactivity between both helodermatid lizard isolates. However, squamate plasma showed different reactions to each of these lizard isolates in neutralisation tests.

Molecular identification of Sarcocystis spp. helped to define the origin of green pythons (Morelia viridis) confiscated in Germany.

Sarcocystis spp. represent apicomplexan parasites. They usually have a heteroxenous life cycle. Around 200 species have been described, affecting a wide range of animals worldwide, including reptiles. In recent years, large numbers of reptiles have been imported into Europe as pets and, as a consequence, animal welfare and species protection issues emerged. A sample of pooled feces from four confiscated green pythons (Morelia viridis) containing Sarcocystis spp. sporocysts was investigated. These snakes were imported for the pet trade and declared as being captive-bred. Full length 18S rRNA genes were amplified, cloned into plasmids and sequenced. Two different Sarcocystis spp. sequences were identified and registered as Sarcocystis sp. from M. viridis in GenBank. Both showed a 95-97% sequence identity with the 18S rRNA gene of Sarcocystis singaporensis. Phylogenetic analysis positioned these sequences together with other Sarcocystis spp. from snakes and rodents as definitive and intermediate hosts (IH), respectively. Sequence data and also the results of clinical and parasitological examinations suggest that the snakes were definitive hosts for Sarcocystis spp. that circulate in wild IH. Thus, it seems unlikely that the infected snakes had been legally bred. Our research shows that information on the infection of snakes with Sarcocystis spp. may be used to assess compliance with regulations on the trade with wildlife species.