Discovery of a new Hepatozoon species namely Hepatozoon viperoi sp. nov. in nose-horned vipers in Türkiye.

Although Hepatozoon spp. remains the most prevalent intracellular protozoa infecting snakes, it was reported only in a few snake species of the Colubridae family in Türkiye. Moreover, studies on these hemoparasites are not available in venomous nose-horned vipers from Türkiye. In this study, we investigated Hepatozoon spp. in three individual Vipera ammodytes using morphological and molecular methods. Our results were positive for intraerythrocytic Hepatozoon spp. gamonts in all three snakes, exhibiting low parasitemia. The microscopic findings were further confirmed through molecular data. A genus-specific PCR assay targeting the 18S rRNA gene region of Hepatozoon spp., was performed using HemoF/HemoR and Hep300/Hep900 primers. The obtained sequences were concatenated and used in phylogenetic analyses in comparison with different Hepatozoon species. Although our (OP377741) isolate was separated into a different branch, it was clustered with the isolates of H. massardi (KC342526), H. cevapii (KC342525), and H. annulatum (ON262426) from Brazilian snakes. Moreover, gene similarity and pair-wise distance between our isolate and other Hepatozoon species infecting snakes were found to be 89.30-98.63% and 0.009-0.077, respectively. Hence, we reported a new species of Hepatozoon, namely Hepatozoon viperoi sp. nov. infecting V. ammodytes. Since the literature does not indicate the existence of such a Hepatozoon species in V. ammodytes in different countries, our data may contribute to the expanding knowledge of Hepatozoon species in snakes, providing new insights into the biodiversity of the haemogregarine protozoan parasite.

Prevalence of Haemogregarina stepanowi and Assessment of Some Risk Factors in Mauremys rivulata (Valenciennes, 1833) Freshwater Turtles (Testudines: Geoemydidae)

Objective: The Balkan terrapin, Mauremys rivulata, is a freshwater turtle. This reptile is exposed to many environmental pollutants and some infectious agents, including Haemogregarina stepanowi parasite. This study was conducted to determine the microscopic prevalence of haemogregarine infection in M. rivulata caught in three different localities (Bozcaada, Gökçeada, and Dardanos) in Çanakkale province of Turkey, and assessment of some risk factors. Methods: Twenty-four blood samples were collected, thin blood smears were prepared, and the presence of haemogregarine parasites microscopically was screened. Water samples were also taken from the habitats, and these samples were analyzed physiochemically and microbiologically. Results: Morphological identification was made by detecting the sausage-shaped intra-cytoplasmic developmental stages of H. stepanowi, and thirteen of twenty-four turtles (54.2%) were found to be infected. The prevalence of H. stepanowi was detected as the highest (90.0%) in the Gökçeada district, where the water pollution is higher than in the other localities. A statistically significant relationship was observed between the distribution of the infection and the gender of the turtles, the temperature of the water, the number of faecal coliforms in water and the amount of dissolved oxygen in the water. A statistically significant difference was found between the localities in terms of the prevalence of H. stepanowi infection, and the infection was primarily detected in the Gökçeada district. Conclusion: This study has significance in providing information regarding haemoparasitic diseases of freshwater turtle, M. rivulata, in Turkey.

Determination of the effects of environmental pollution on the Balkan terrapin, Mauremys rivulata (Valenciennes, 1833).

The effects of environmental pollution on three populations of the Balkan terrapin [Mauremys rivulata (Valenciennes, 1833)] from the Bozcaada, Gökçeada and Dardanos regions were evaluated. The morphological parameters of Balkan terrapins collected on each site were measured and blood samples were taken for haematological analysis and micronucleus detection. The physicochemical, microbiological and microelement analyses of the water samples from each region were conducted by standard methods. The highest red blood cell, white blood cell and mean corpuscular haemoglobin concentration values were seen in the samples from Gökçeada. The highest haemoglobin value was found in the samples originating from Bozcaada, whereas the highest haematocrit and mean corpuscular volume values were found in the animals from Dardanos. Based on the microbiological analysis of the water samples, the most polluted site was Gökçeada. The microelement contents of the water and blood samples were different at the three sites, the lowest being in the Gökçeada area. It was revealed that the percentage of red blood cell micronuclei and other nucleus abnormalities in the M. rivulata blood samples was the lowest also in the animals living in the region of Gökçeada.

Determination of antimicrobial and heavy metal resistance profiles of some bacteria isolated from aquatic amphibian and reptile species.

The aim of the present study was to determine the level of antibiotic resistance patterns and distribution of heavy metal resistance of bacterial isolates from aquatic animals (Lissotriton vulgaris, Pelophylax ridibundus, Emys orbicularis, Mauremys rivulata, and Natrix natrix) in Turkey (Kavak Delta). A total of 153 bacteria have been successfully isolated from cloaca and oral samples of the aquatic amphibians and reptilians which were found, namely, Aeromonas sp. (n = 29), Plesiomonas sp. (n = 7), Vibrio sp. (n = 12), Citrobacter sp. (n = 12), Enterobacter sp. (n = 11), Escherichia sp. (n = 22), Klebsiella sp. (n = 22), Edwardsiella sp. (n = 6), Hafnia sp. (n = 1), Proteus sp. (n = 19), Providencia sp. (n = 8), and Pseudomonas sp. (n = 4). In terms of antibiotic and heavy metal susceptibility testing, each isolate was tested against 12 antibiotics and 4 metals. There was a high incidence of resistance to cefoxitin (46.40 %), ampicillin (44.44 %), erythromycin (35.29 %), and a low incidence of resistance to gentamicin (6.53 %), kanamycin (8.49 %), chloramphenicol (9.15 %), and cefotaxime (10.45 %). The multiple antibiotic resistance index of each bacterial species indicated that bacteria from raised amphibians and reptiles have been exposed to tested antibiotics, with results ranging from 0 to 0.58. Most isolates showed tolerance to different concentrations of heavy metals, and minimal inhibition concentrations ranged from100 to >3,200 µg/mL. According to these results, a significant occurrence of bacteria in the internal organs of reptiles and amphibians, with a high incidence of resistance against antibiotics and heavy metals, may risk aquatic animals and the public health. These data appoint the importance of epidemiological surveillance and microbiological monitoring and reinforce the need to implement environment protection programs for amphibian and reptile species.