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.

Are owned dogs or stray dogs more prepared to diseases? A comparative study of immune system gene expression of perforin and granzymes.

Stray dogs are inevitably exposed to more infections than owned dogs living indoor. However, no studies have investigated whether the immune system gene expression differs between owned dogs kept in better care conditions and stray dogs living outside. To investigate this, blood samples were taken from 90 dogs (45 owned and 45 stray dogs) that were checked and confirmed as healthy. By using qPCR, the amples were analyzed for the expression of the perforin, granzyme A and granzyme B genes, which are associated with the activation of apoptotic pathways in the immune system. We found that the perforin and granzyme A gene expression levels were higher in stray dogs although the differences were not statistically significant. On the other hand, a 2.81 times higher and a notable difference (P < 0.001) was found in the expression level of granzyme B gene in stray dogs. These results indicate that the immune system in stray dogs might be more prepared for diseases than that of the owned dogs and the granzyme B gene plays a more dominant role in the immune response than granzyme A and perforin.

Comparison of microbiota and volatile organic compounds in milk from different sheep breeds.

In this study, we compared the microbiota and volatile organic compounds (VOC) present in the milk obtained from 3 different sheep breeds, namely Merino, Lacaune, and Assaf. Udder milk was collected from 21 animals, 7 from each breed. Bacterial microflora was determined metagenomically by extracting the DNA from the milk and analyzing the V3-V4 region of the 16S rRNA gene. Headspace solid-phase microextraction gas chromatography-mass spectrometry method was used to analyze VOC. The metagenomic analysis revealed (for Merino, Lacaune, and Assaf milk, respectively) Firmicutes (66.32, 69.36, and 57.08%), Actinobacteria (19.09, 7.67, and 19.40%), Proteobacteria (13.76, 21.06, and 22.19%), and Bacteroidetes (0.84, 1.91, and 1.33%) phyla in the milk samples. Lactobacillus was highly abundant in the milk of 3 breeds (29.64, 43.50, and 18.70%). The genera constituting more than 2% of all bacteria in all groups were Jeotgalicoccus (7.19, 5.34, and 10.77%), Enterococcus (5.18, 9.78, and 3.64%), and Corynebacterium (4.08, 3.00, and 13.44%). A total of 32 different VOC were identified by headspace solid-phase microextration analysis with 9, 30, and 24 different compounds from Merino, Lacaune, and Assaf breeds, respectively. Although ketone was the most abundant compound in Merino milk (71.84%), hydrocarbons were the most detected in Lacaune and Assaf milk (37.18% and 55.42%, respectively). A positive correlation was found between acetone, which was detected at the highest level in all groups, with Salinicoccus, Alloiococcus, Psychrobacter, and Dietzia. In addition, a negative correlation was found between the Lactobacillus genus, detected at the highest level in all groups, with methyl cyclopentane, 3-methylheptane, octane, decane, 3,3-dimethyloctane, and dodecane. Thus, differences were observed in the bacterial microflora and VOC in the sheep milk from different breeds under different feeding and breeding conditions.

Ectoparasites of feral horses [Equus ferus caballus (Linnaeus., 1758)] on Karadag Mountain, Karaman, Turkey.

Approximately 250 feral horses [Equus ferus caballus (Linnaeus, 1758)] living on Karadag Mountain near Karaman City were caught by Kazakh horse herdsmen with permission of the Turkish Ministry of Agriculture and Forestry and brought to a farm in Karkin village in Konya Province, 70 km from Karadag, in November, 2017. This study was carried out to determine the presence of ectoparasites infesting a subsample of 36 feral horses. The horses were visually inspected, and then their bodies were checked by hand for ectoparasites. Thirty-five (97.2%) were infested with at least one of five species of ectoparasites: Bovicola equi (Linnaeus, 1758), Hippobosca equina (Linnaeus, 1758), Haemaphysalis parva (Neuman, 1897), Hyalomma excavatum (Koch, 18449), Dermacentor marginatus (Sulzer, 1776). Most of the horses were coinfested with two ectoparasite species. Prevalence of infestation with H. equina was 80.6% and with B. equi 72.2%. In addition, prevalence of Ha. parva was 25.0%, Hy. excavatum 13.9%, and D. marginatus was 5.6%. This is the first systematic examination for external parasites of feral horses in Turkey. Further studies are needed to determine ectoparasites of greater numbers of feral horses in different localities.

Sulfasalazine treatment can cause a positive effect on LPS-induced endotoxic rats.

The aim of this study, was to determine the effect of sulfasalazine for different periods of time reduces disseminated intravascular coagulation, inflammation and organ damages by inhibiting the nuclear factor kappa beta pathway. The study was performed with 30 Wistar albino rats and the groups were established as Control group, LPS group; endotoxemia was induced with LPS, SL5 group: sulfasalazine (300 mg/kg, single dose daily) was administered for 5 days before the LPS-induced endotoxemia, and LS group: sulfasalazine (300 mg/kg, single dose) was administered similtenously with LPS. Hemogram, biochemical, cytokine (IL-1ß, IL-6, IL-10, TNF-α) and acute phase proteins (HPT, SAA, PGE2) analyzes and oxidative status values were measured from blood samples at 3 and 6 h after the last applications in the all groups. The rats were euthanized at 6 h and mRNA levels of BCL2 and BAX genes were examined from liver and brain tissues. Sulfasalazine reduced the increased IL-1ß, IL-6, TNF-α and PGE2 levels and significantly increased anti-inflammatory cytokine IL-10 levels. In addition, decreasing of ATIII level was prevented in the SL5 group, and decreasing of fibrinogen levels were prevented in the LS and SL5 groups within first 3 h. In LPS group, leukocyte and thrombocyte levels were decreased, however sulfasalazine application inhibited decreases of leukocyte levels in LS and SL5 groups. In addition, sulfasalazine inhibited the decrease of total antioxidant capacity and unchanged apoptosis in brain and liver. In conclusion, the use of sulfasalazine in different durations reduce the excessive inflammation of endotoxemia cases.