Capillary Zone Electrophoresis in Humboldt Penguins (Spheniscus humboldti).

Electrophoresis is a useful diagnostic tool for detecting inflammation, including inflammation associated with infectious diseases (eg, aspergillosis in penguins). To our knowledge, reference intervals are not available for plasma proteins via electrophoresis in Humboldt penguins (Spheniscus humboldti). Therefore, preliminary reference intervals for blood plasma proteins measured by capillary zone electrophoresis were calculated for Humboldt penguins from a single zoological collection, and possible differences between the sexes and the ages of the birds were evaluated. Lithium heparinized plasma samples from 39 Humboldt penguins were analyzed. The following sex- and age-independent reference intervals were calculated: total protein 33.8-70.4 g/L, prealbumin 1.9-4.9 g/L, albumin 12.9-31.1 g/L, albumin: globulin ratio 0.7-1.7, α-globulins 4.5-11.6 g/L, ß-globulins 5.6-20.6 g/L, and γ-globulins 2.6-8.4 g/L. Male penguins had a significantly (P = 0.047) higher albumin: globulin ratio and lower percentage of ß-globulins (P = 0.015) in comparison with female penguins. Prealbumin (g/L) significantly (P = 0.021) decreased with increased age of the penguins. These results showed some differences between the sexes and ages of the penguins, which should be considered when interpreting the results. Further studies are needed to determine whether differences in other age groups or seasons exist, and also to evaluate which infectious diseases affect plasma proteins and how the reference values calculated here may deviate in ill penguins.

Clinical Chemistry and Haematology Values of a Captive Population of Humboldt Penguins (Spheniscus humboldti).

Reference intervals for clinical chemistry and haematology analytes are essential for the interpretation of blood results, but limited data are available for Humboldt penguins (Spheniscus humboldti) in zoological collections as well as in the wild. The aim of the present study was therefore to establish reference intervals for a series of clinical chemistry and haematology analytes in a single zoological population of Humboldt penguins. Furthermore, possible variations of the analytes influenced by the age and sex of the individuals were investigated. Lithium heparinized plasma and whole blood samples from 39 animals were analysed and statistically evaluated. The following clinical chemistry analytes were significantly higher in females: glutamate dehydrogenase (p = 0.0003), alanine transaminase (p = 0.0005), alkaline phosphatase (p = 0.0245), aspartate aminotransferase (p = 0.0043), creatine kinase (p = 0.0016), lactate dehydrogenase (p < 0.0001), inorganic phosphorus (p = 0.0223), and sodium (p = 0.0415). No significant differences between males and females were found for any haematology analyte. Cholesterol (p = 0.0233; -0.39), white blood cell count (p = 0.0208; -0.40), and absolute heterophiles (p = 0.0148; -0.42) had significant negative correlations with the age of the penguins. The results indicate that the factors of sex and age should be considered when interpreting the results of haematological and clinical biochemistry assays, and further studies are needed to determine whether there are more differences in other age groups or seasons.

Salmonella in reptiles: a review of occurrence, interactions, shedding and risk factors for human infections.

Salmonella are considered a part of the normal reptile gut microbiota, but have also been associated with disease in reptiles. Reptile-associated salmonellosis (RAS) can pose a serious health threat to humans, especially children, and an estimated 6% of human sporadic salmonellosis cases have been attributed to direct or indirect contact with reptiles, although the exact number is not known. Two literature searches were conducted for this review. The first evaluated reports of the prevalence of Salmonella in the intestinal tracts of healthy reptiles. Salmonella were most commonly detected in snakes (56.0% overall), followed by lizards (36.9%) and tortoises (34.2%), with lower detection rates reported for turtles (18.6%) and crocodilians (9%). Reptiles in captivity were significantly more likely to shed Salmonella than those sampled in the wild. The majority of Salmonella strains described in reptiles belonged to subspecies I (70.3%), followed by subspecies IIIb (29.7%) and subspecies II (19.6%). The second literature search focused on reports of RAS, revealing that the highest number of cases was associated with contact with turtles (35.3%), followed by lizards (27.1%) and snakes (20.0%). Reptiles associated with RAS therefore did not directly reflect prevalence of Salmonella reported in healthy representatives of a given reptile group. Clinical symptoms associated with RAS predominantly involved the gastrointestinal tract, but also included fever, central nervous symptoms, problems with circulation, respiratory symptoms and others. Disease caused by Salmonella in reptiles appears to be dependent on additional factors, including stress, inadequate husbandry and hygiene, and other infectious agents. While it has been suggested that reptile serovars may cause more severe disease than human-derived strains, and some data is available on invasiveness of individual strains in cell culture, limited information is available on potential mechanisms influencing invasiveness and immune evasion in reptiles and in RAS. Strategies to mitigate the spread of Salmonella through reptiles and to reduce RAS focus mostly on education and hygiene, and have often been met with some success, but additional efforts are needed. Many aspects regarding Salmonella in reptiles remain poorly understood, including the mechanisms by which Salmonella persist in reptile hosts without causing disease.

The Role of Host Species in Experimental Ferlavirus Infection: Comparison of a Single Strain in Ball Pythons (Python regius) and Corn Snakes (Pantherophis guttatus).

Ferlaviruses are a cause of respiratory disease in snakes. Four genogroups (A, B, C, and tortoise) have been described. Disease development is believed to depend on virus, host, and environment-specific factors. There is evidence of transmission of individual strains between genera and families of reptiles. A genogroup B virus previously used in a transmission study with corn snakes (Pantherophis guttatus) was applied intratracheally in ball pythons (Python regius) using the same protocol as for the corn snakes. Ball pythons became infected, with initial mild clinical signs noted four days post infection (p.i.), and the virus was detected first in the lungs on day 4 and spread to the intestine, pancreas, kidney and brain. Hematology showed an increase in circulating lymphocytes which peaked on day 28 p.i. Antibodies were detected beginning on day 16 and increased steadily to the end of the study. In comparison to corn snakes, ball pythons exhibited milder clinical signs and pathological changes, faster development of and higher antibody titers, and a hematological reaction dominated by lymphocytosis in contrast to heterophilia in corn snakes. These differences in host reaction to infection are important to understand ferlavirus epidemiology as well as for clinical medicine and diagnostic testing.

Comparison of Plasma Trace Elements Between 2 Captive Humboldt Penguin (Spheniscus humboldti) Populations.

Environmental pollutants and their effect on wildlife health play an important role in the conservation of endangered species and can be clinically relevant in captive animals too. Data on relevant concentrations of trace elements in captive birds with no known exposures are rare. For this study, silver, arsenic, gold, barium, cadmium, cobalt, chromium, copper, iron, mercury, magnesium, manganese, molybdenum, thallium, selenium, and zinc were measured by inductively coupled plasma mass spectrometry in lithium heparinized plasma samples from 18 Humboldt penguins at 2 zoological collections in Germany. The results showed that the plasma concentrations of silver, arsenic, gold, cadmium, cobalt, chromium, mercury, and selenium differed significantly (P ≤ 0.05) between the 2 penguin collections. The results indicate that the location of the birds has a strong influence on the plasma trace element concentrations. Well water used in the enclosures was suspected to be associated with these significant differences. Trace elemental concentrations in feed (eg, marine fish) and contamination from enclosure construction materials may also play a role. This study could provide a basis for further comparative, biomonitoring, toxicity, and reference interval studies.

Establishment of a Real-Time PCR Assay for the Detection of Devriesea agamarum in Lizards.

(1) Background: Devriesea (D.) agamarum is a potential cause of dermatitis and cheilitis in lizards. The aim of this study was to establish a real-time PCR assay for the detection of D. agamarum. (2) Methods: Primers and probe were selected targeting the 16S rRNA gene, using sequences of 16S rRNA genes of D. agamarum as well as of other bacterial species derived from GenBank. The PCR assay was tested with 14 positive controls of different D. agamarum cultures as well as with 34 negative controls of various non-D. agamarum bacterial cultures. Additionally, samples of 38 lizards, mostly Uromastyx spp. and Pogona spp., submitted to a commercial veterinary laboratory were tested for the presence of D. agamarum using the established protocol. (3) Results: Concentrations of as low as 2 × 104 colonies per mL were detectable using dilutions of bacterial cell culture (corresponding to approximately 200 CFU per PCR). The assay resulted in an intraassay percent of coefficient of variation (CV) of 1.31% and an interassay CV of 1.80%. (4) Conclusions: The presented assay is able to detect D. agamarum in clinical samples, decreasing laboratory turn-around time in comparison to conventional culture-based detection methods.

Genetic diversity among reptilian orthoreoviruses isolated from pet snakes and lizards.

Reovirus infections in reptiles are frequently detected and associated with various clinical diseases; yet, our knowledge about their genetic diversity and evolutionary relationships remains limited. In this study, we characterize at the genomic level five reptile origin orthoreovirus strains isolated from exotic snakes and lizards in Hungary and Germany. The genomic organization of the study strains was similar to that of the representative strains of reptile origin reoviruses belonging to species Reptilian orthoreovirus and Testudine orthoreovirus. Additionally, all five study strains clustered with the bush viper origin reference Reptilian orthoreovirus strain, 47/02. The nucleotide sequence divergence among strains fell from 56.64 to 99.36%. Based on genome segment constellations two well separated groups were observed, which may represent two genetic lineages of reptilian orthoreoviruses we tentatively referred here as genogroups, classifying two squamata origin strains with available whole genome sequences into genogroup I (GGI) and four strains into genogroup II (GGII). The representative GGI and GGII Reptilian orthoreovirus strains are characterized by moderate-to-high nucleotide and amino acid similarities within genogroups (range, 69.45 to 99.36% and 74.64 to 100.00%), whereas lower nucleotide and amino acid similarities (range, 56.64 to 77.24% and 54.53 to 93.85%) and different structures of the bicistronic S1 segment were found between genogroups. Further studies are needed to explore the genomic diversity among reptilian reoviruses of squamata origin; this would be critical to establish a robust classification system for these viruses and to see if interaction among members of distinct lineages may result in viable progenies with novel genetic features.

Detection of nidoviruses in samples collected from captive snakes in Europe between 2016 and 2021.

BACKGROUND: Nidoviruses are increasingly detected in various snake species worldwide, but much remains to be learned about their distribution and the factors influencing their epidemiology. METHODS: This retrospective study evaluated the results of routine nidovirus testing, by PCR, of 5210 swab samples from pet snakes from various European countries that were submitted to a commercial veterinary laboratory in Germany between 2016 and 2021. RESULTS: The overall detection rate was 19.96%. However, the detection rate varied significantly depending on the snake species (p < 0.0001), with the highest rate in Indian pythons (Python molurus) (42.24%). Rates also varied depending on the season of sample collection (p < 0.0001), with the highest rate in winter (24.46%), and the country of sample origin (p < 0.0001), with the highest rate in Austria (36.69%). The detection rate also decreased significantly (p = 0.0003) over the 6-year observation period, from 26.43% to 17.64%. LIMITATION: No information on clinical signs was available for most of the sampled snakes. CONCLUSION: The present study supplies new information on the distribution of python nidoviruses (subgenus Roypretovirus) in pet snakes in Europe and indicates a dynamic situation with possible changes in prevalence over time.

Infectious disease threats to amphibians in Greece: new localities positive for Batrachochytrium dendrobatidis.

In the early 2000s, numerous cases of European amphibian population declines and mass die-offs started to emerge. Investigating those events led to the discovery that wild European amphibians were confronted with grave disease threats caused by introduced pathogens, namely the amphibian and the salamander chytrid fungi Batrachochytrium dendrobatidis (Bd) and B. salamandrivorans (Bsal) and ranaviruses. In Greece, Bd was previously documented among wild amphibian populations in 2 different locations and 3 different species. However, no disease-related mass declines or mortality events have been reported. In this work, we build upon previous findings with new, subsequently obtained data, resulting in a 225-sample dataset of 14 species from 17 different locations throughout Greece, in order to examine the occurrence status of all 3 pathogens responsible for emerging infectious diseases in European amphibians. No positive samples for Bsal or ranavirus were recorded in any location. We confirmed the presence of Bd in 4 more localities and in 4 more species, including 1 urodelan (Macedonian crested newt Triturus macedonicus) and 1 introduced anuran (American bullfrog Lithobates catesbeianus). All insular localities were negative for Bd, except for Crete, where Bd was identified in 2 different locations. Again, no mass declines or die-offs were recorded in any Bd-positive area or elsewhere. However, given the persistence of Bd across Greece over the past ~20 yr, monitoring efforts should continue, and ideally be further expanded.

PLASMA VITAMIN AND MINERAL CONCENTRATIONS IN CAPTIVE GREATER FLAMINGOS (PHOENICOPTERUS ROSEUS) AS INFLUENCED BY DIET CHANGE.

The greater flamingo (Phoenicopterus roseus) is often maintained in managed care. Although values for plasma concentrations of minerals and vitamins have been published for this species, limited studies investigate the effects of diet changes on these values. Plasma concentrations of selected vitamins (A [and the provitamin A carotenoid ß-carotene] and E), minerals (Ca, Cu, Fe, K, Mg, Mn, Na, inorganic P, Se, and Zn), and total protein (TP) were measured in a zoo collection of greater flamingos in November 2018 (n = 48) and in November 2019 (n = 52), 30 d after a change from the summer diet, which was the same in both years, to different winter diets. Chloride, cholesterol, and triglycerides were only measured once in 2019. The nutrient profiles of the two complete diets (labeled and analyzed) differed considerably, especially for the canthaxanthin concentration. The plasma concentrations of ß-carotene (P < 0.0001), Cu (P < 0.0001), Fe (P < 0.0001), Mg (P < 0.0001), and Se (P < 0.0001) were significantly higher and vitamin E (P < 0.0001), Ca (P = 0.0014), Mn (P < 0.0001), Na (P = 0.0005), and TP (P < 0.0001) were significantly lower in 2018 than in 2019. Four minerals (Cu, Fe, Mg, and Zn) were measured using both spectrophotometry and inductively coupled plasma mass spectrometry, and all of these minerals showed significant (P < 0.05) differences between the two methods. Based on the results of the study, both the influence of diet and the detection method used for circulating nutrients must be considered when interpreting the plasma vitamin and mineral concentrations in greater flamingos.

Antibodies against Two Testudinid Herpesviruses in Pet Tortoises in Europe.

Herpesviruses are important pathogens of tortoises, and several serologically and genetically distinct virus types have been described in these animals. Virus neutralization testing is commonly used in Europe to determine previous infection with the two types most often found in pet European tortoises, testudinid herpesvirus (TeHV) 1 and 3. In this retrospective study, the results of serological testing for antibodies against each of these viruses in serum or plasma samples from 1728 tortoises were evaluated, and antibody detection rates were compared based on virus type, host species, year, season, and country of origin. Antibodies (titer 2 or higher) against at least one of the two viruses used were detected in a total of 122 (7.06%; 95% CI 5.95−8.37%) of the animals tested. The antibody detection rates differed significantly depending on the tortoise species (p < 0.0001) and the year of sampling (TeHV1 p = 0.0402; TeHV3 p = 0.0482) for both virus types. For TeHV1, antibody detection rates differed significantly (p = 0.0384) by season. The highest detection rate was in summer (5.59%; 95% CI 4.10−7.58%), and the lowest was in fall (1.25%; CI 0.53−2.87%). TeHV1 antibody detection rates did not differ significantly (p = 0.7805) by country, whereas TeHV3 antibody detection rates did (p = 0.0090). The highest detection rate, 12.94% (95% CI 7.38−21.70%), was found in samples from Italy. These results support previous hypotheses on the species' susceptibility to TeHV1 and 3 and the use of serology as a diagnostic test for the detection of herpesvirus-infected tortoises.

COMPARISON OF CLINICAL CHEMISTRY ANALYTES IN GREATER FLAMINGOS (PHOENICOPTERUS ROSEUS) AND AMERICAN FLAMINGOS (PHOENICOPTERUS RUBER) IN A ZOO IN EUROPE.

Reference intervals for clinical chemistry analytes are important for the interpretation of blood results, but reference intervals available in journal articles for specific flamingo collections are often based on a small number of individuals or are only available for a limited number of analytes. Differences in husbandry and nutritional conditions may impact reference intervals. The goal of this study was to establish reference intervals from a mixed population of greater (Phoenicopterus roseus) and American (Phoenicopterus ruber) flamingos when husbandry parameters are equal and to compare the results between the different species, sexes, and age groups in order to evaluate differences between these factors. Lithium-heparinized plasma samples from 93 animals were analyzed for the following: amylase, cholinesterase (CHE), bile acids (BA), total protein (TP), glutamate dehydrogenase (GLDH), aspartate aminotransferase (AST), creatine kinase (CK), lactate dehydrogenase (LDH), uric acid (UA), inorganic phosphorus (P), total calcium (Ca), sodium (Na), and potassium (K). The results show many differences between the species and a few between the different age groups but no differences between males and females of either species. Amylase (P= 0.0010), AST (P= 0.0090), CK (P= 0.0011), and Ca (P= 0.0217) differed significantly between greater and American flamingos. The age group significantly impacted blood levels of CK (P = 0.0299) and LDH (P = 0.0165) in greater flamingos and BA (P = 0.0495), P (P = 0.0422), and Ca (P = 0.0006) in American flamingos. The results show the importance of species-specific reference intervals and the consideration of age-related variations in order to facilitate correct interpretation of blood results.

A testudinid herpesvirus 1 (TeHV1)-associated disease outbreak in a group of Horsfield's tortoises (Testudo horsfieldii).

In spring 2020, a severe disease outbreak with high morbidity and mortality was observed in a collection of 15 Horsfield's tortoises (Testudo horsfieldii). Affected tortoises showed upper respiratory- and gastrointestinal tract signs, including rhinitis and stomatitis. Testudinid herpesvirus 1 (TeHV1) and Mycoplasma spp. were detected by PCR in oral swabs of affected animals. Histological examination of one deceased animal showed intranuclear inclusion bodies typical for herpesvirus infections in liver, spleen and oesophagus. The virus was likely introduced into the collection 2 years earlier by a clinically healthy Horsfield's tortoise that was tested positive for TeHV1 by PCR.

Herpesviruses in Captive Chelonians in Europe Between 2016 and 2020.

Herpesviruses are important pathogens in tortoises and turtles, yet little is known about the epidemiology of these viruses. We analyzed herpesviruses detected by PCR in samples from captive chelonians in Europe according to virus strain, host species, year and season in which the animal was tested, and country in which the animal was kept. A total of 4,797 samples submitted to a diagnostic laboratory in Europe between January 2016 and December 2020 were evaluated. Of these, 312 (6.50%) were positive for herpesviruses. The types most commonly found were testudinid herpesvirus (TeHV)1 (143 positive, 45.83%) and TeHV3 (153 positive, 49.04%), but also included TeHV2 (1 positive, 0.32%), TeHV4 (3 positive, 0.96%), Terrapene herpesvirus 1 (7 positive, 2.24%), Trachemys herpesvirus 1 (2 positive, 0.64%), and three previously undescribed herpesviruses (0.96%). Herpesviruses were detected in chelonians in the families Testudinidae, Emydidae, Geoemydidae, and in the suborder Pleurodira. Among the species for which 100 samples or more were available, the highest proportions of positive samples (positivity rates) were found in samples from Horsfield's tortoises (Testudo horsfieldii) (14.96%), and radiated tortoises (Astrochelys radiata) (14.05%). Among tortoises (Testudinidae), viruses were most often detected in the spring, while in emydid turtles (Emydidae) they were most often detected in the summer. A comparison of the positivity rates according to country showed significant differences, with the highest rate in samples from Italy (16.01%). This study indicated possible differences in herpesvirus positivity rates depending on host species, virus strain, year of sampling, season, and country of origin. It provides useful information in further understanding fluctuations in infection rates as well as in helping to guide decision making for herpesvirus diagnostics in chelonian patients. It also provides evidence for the international dispersal of herpesviruses with their hosts through international trade.

Two Novel Adenoviruses in Free-Living British lizards.

In this study, we describe two novel adenoviruses isolated from (i) a common lizard (Zootoca vivipara) found dead and examined post-mortem and (ii) pooled samples from free-living sand lizards (Lacerta agilis agilis). Sequencing indicated the two were closely related atadenovirus strains which were distinct from previously recorded adenoviruses in lizards. Adenoviruses are not always associated with disease in squamates, but morbidity and mortality have been reported. These are the first known cases of adenovirus infection in free-living native British lizards, and further monitoring will be necessary to elucidate the implications of these possible pathogens for vulnerable populations of native reptiles.

Variation in mineral types of uroliths from ferrets (Mustela putorius furo) submitted for analysis in North America, Europe, or Asia over an 8-year period.

OBJECTIVE: To compare mineral types of naturally occurring uroliths in ferrets (Mustela putorius furo) from North America, Europe, and Asia and to identify potential risk factors associated with cystine urolithiasis in ferrets. SAMPLES: 1,054 laboratory submission records of uroliths obtained from ferrets between January 1, 2010, and December 31, 2018. PROCEDURES: For this cross-sectional study, the medical records databases at 4 diagnostic laboratories were searched for records of submissions of uroliths obtained from ferrets. Data collection included submission date; ferret sex, neuter status, and age; receiving laboratory and continent; and urolith mineral type. Regression analyses were performed to identify variables associated with cystine uroliths. RESULTS: Of the 1,054 urolith submissions, 1,013 were from North America, with 92.6% (938/1,013; 95% CI, 90.8% to 94.1%) cystine uroliths, and 41 were from Europe and Asia, with only 26.8% (11/41; 95% CI, 15.7% to 41.9%) cystine uroliths. Median age was 2.0 years for ferrets with cystine urolithiasis versus 4.0 years for those with other types of uroliths. Submissions were more likely cystine uroliths for ferrets in North America versus Europe and Asia (adjusted OR [aOR], 59.5; 95% CI, 21.4 to 165.6), for ferrets that were younger (aOR, 0.67; 95% CI, 0.58 to 0.77), or for submissions in 2018 versus 2010 (aOR, 21.1; 95% CI, 5.1 to 87.9). CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that the proportion of submissions that were cystine uroliths dramatically increased in North America between 2010 and 2018. There is an urgent need to determine underlying causes and mitigate cystine urolithiasis in ferrets.

Comparison of Capillary Zone Electrophoresis in Greater Flamingos (Phoenicopterus roseus) and American Flamingos (Phoenicopterus ruber).

Electrophoresis can be used to aid in the diagnosis of infectious diseases (eg, aspergillosis) in avian species. Reference intervals for blood plasma proteins of 2 different flamingo species (Phoenicopterus roseus and Phoenicopterus ruber) and their hybrids were calculated by capillary zone electrophoresis (CZE) and differences between these species, sexes, and age groups were evaluated. Lithium-heparinized plasma samples from 111 animals from a zoological collection were analyzed by CZE and statistically evaluated. Differences were only found between greater and American flamingos (P = .003) and between greater flamingos and hybrids (P = .001) in the γ-globulin fraction. Male greater flamingos showed significantly higher α-globulins (P = .022) and females higher total albumin by CZE (P = .037). In American flamingos, the percent total albumin (P = .017), total albumin (P = .025), prealbumin (P = .005), and albumin/ globulin (A/G) ratio (P = .008) were higher in females, and α- (P = .023) and ß-globulins (P = .021) were higher in males of the same species. The following parameters differed significantly between the age groups: γ-globulins (P = .048) in greater and α- (P = .021) and ß-globulins (P = .001) in American flamingos increased with increased age and percent total albumin (P = .002), total albumin (P = .024), and A/G ratio (P = .002) decreased with age in American flamingos. The results showed only small differences between the species, but greater differences between the sexes and ages, especially in American flamingos, which must be considered when interpreting laboratory results.

A NOVEL IGUANID HERPESVIRUS DETECTED IN ASYMPTOMATIC GREEN IGUANAS (IGUANA IGUANA) IN POLAND.

While herpesviruses are well-known pathogens in a wide variety of chelonian species, they have only sporadically been documented in squamate reptiles. Those that have been described have most often been associated with hepatic disease and oral lesions. During a study on infectious disease in pet reptiles in Poland, herpesviruses were detected in swabs from three green iguanas (Iguana iguana) from two different owners that were presented to two different veterinary clinics in Warsaw. One iguana was presented for abscesses on the head, while the other two were partner animals and remained clinically healthy throughout the course of this study. Virus was detected in oral swabs as well as combined swab samples from the oral cavity and cloaca using a panherpesvirus PCR. PCR products from all three animals were sequenced, and the detected viruses were most closely related to iguanid herpesvirus 2 from a San Esteban chuckwalla (Sauromalus varius) in the United States (GenBank accession No. AY236869.1). The single animal was retested again 1 y later and remained clinically healthy and continued to shed the same herpesvirus. This is the first description of a herpesvirus infection in pet iguanas in Europe. While the clinical relevance of the infection is not known, it is of interest that the infected animals appeared to continue to shed virus over an extended period of time.

DETECTION OF TESTADENOVIRUSES AND ATADENOVIRUSES IN TORTOISES AND TURTLES IN EUROPE.

Adenoviruses have been regularly detected in squamate reptiles; evidence of infection in chelonians is described much less frequently. The adenoviruses found in turtles and tortoises have been genetically diverse, and have included members of the genus Siadenovirus, a proposed testadenovirus genus, and, in a single case, an Atadenovirus. In this study, samples from 949 chelonians submitted to a diagnostic laboratory were screened for the presence of adenoviruses by polymerase chain reaction (PCR) targeting a portion of the DNA polymerase gene. Adenoviruses were detected in 22 (2.3%) chelonians of different species. Adenovirus-positive species included Hermann's tortoises (Testudo hermanni), spur-thighed tortoises (T. graeca), Horsfield's tortoises (T. horsfieldii), sliders (Trachemys spp.), box turtles (Terrapene spp.) and a black pond turtle (Geochlemys hamiltonii). Sequencing and phylogenetic analyses of the obtained PCR products revealed that the majority of the detected adenoviruses (72.7%) cluster with members of the proposed testadenovirus genus, while the rest (27.3%) cluster with the atadenoviruses. This study significantly expands the known host range of both the proposed testadenoviruses and the atadenoviruses in different chelonian species and families.