Veterinary Research Forum

Abstract This study, aimed to investigate Chlamydia abortus, C. pecorum, and C. psittaci in bovine, sheep, and goat abortions sent to the Konya Veterinary Control Institute diagnostic laboratory from various provinces in Central Anatolia and Mediterranean regions of Turkey between 2019 and 2022 via pathological and molecular methods. For this purpose, a total number of 150 abortions (from 58 cattle, 73 sheep, and 19 goats) were analysed via quantitative real-time polymerase chain reaction, histopathology and immunohistochemistry. The chlamydial DNA positivity rate was determined to be 24.00% via quantitative polymerase chain reaction analyses. According to the animal species, the positivity rate was 45.20% in sheep and 15.78% in goats. No cases positive for chlamydial DNA were found in the cattle abortions analyzed in the present study. When chlamydial agents were analysed on a species basis, C. abortus and C. pecorum were detected in 22.66and 1.33% of the cases, respectively. Among the animal species, 42.47% were C. abortus positive, whereas, 2.74% were C. pecorum positive in sheep. In the present study, the percentage of C. abortus-positive goats was 15.78%. and 22.66% of the Chlamydia spp. were positive according to immunohistochemical staining of aborted fetal lung, liver, heart, placenta and umbilical cord samples. This comparative study revealed that C. abortus was the most common chlamydial abortion agent in Türkiye. Chlamydial abortion agents cause abortion mostly in sheep in the region according to the animal species, and immunohistochemical staining contributes to the diagnosis of the agent at the Chlamydia spp. level.

Abstract This study aimed to obtain a narrow-spectrum antimicrobial peptide. A peptide XMK-8 was designed based on the amino acid sequence of goose MyHC1 protein from positions 1919 to 1936 (some parameters do not meet the requirements of antimicrobial peptides through bioinformatics analysis) using bioinformatics tools and amino acid substitution method. The minimum inhibitory concentration was determined using liquid double dilution method, the hemolysis rate was determined using dilution method, and the effects of temperature, acid-base, enzyme, and salt ions on its antimicrobial activity were evaluated using liquid double dilution method. The results showed that the designed peptide was a cationic hydrophilic peptide with high amphiphilicity and low hemolytic activity on mouse red blood cells. It had no antimicrobial activity against Escherichia coli, Salmonella, Staphylococcus aureus, and Aeromonas hydrophila. The minimum inhibitory concentration against Pasteurella multocida was 250 μg mL^-1, and the minimum inhibitory concentration against Haemophilus parasuis was 1.00 mg mL^-1. The antimicrobial activity of the narrow-spectrum antimicrobial peptide XMK-8 can still be detected after treatment with temperature (0.00 - 100 ˚C), salt ions (sodium ions and potassium ions; 50.00 - 200 mmol L^-1), pH (4.00 - 10.00), and protease K (20.00 - 100 μg mL^-1). Antimicrobial peptide XMK-8 was expected to become a new alternative to antibiotics and would have good application prospects in the prevention and treatment of P. multocida and H. parasuis infections.

Abstract The aim of this study was to investigate the effects of post-partum (PP) periods of different lengths on super-stimulatory and super-ovulatory responses, as well as the number and quality of embryos in Simmental cows. Fifty cows were divided into five groups based on their PP days, including 50 - 60 days (n = 5), 61 - 90 days (n = 17), 91 - 120 days (n = 9), 121 - 150 days (n = 9), and 151 - 420 days (n = 10). On a random day of the estrous cycle, all follicles larger than 5.00 mm on the ovaries were aspirated and a progesterone-releasing device was inserted into the vagina of all donors. Approximately 36 hr after follicle aspiration, all cows were administered 500 µg of follicle-stimulating hormone in decreasing doses, twice daily for 6 days. Ovaries were evaluated using trans-rectal ultrasonography during human chorionic gonadotropin treatment and after embryo collection to assess follicles and corpora lutea, respectively. Super-stimulatory and super-ovulatory responses, as well as embryo yield and quality were found to be similar among cows in the PP days groups of 61 - 90, 91 - 120, 121 - 150, and 151 - 420. However, the embryo recovery rate and mean number of transferable and freezable embryos were adversely affected in cows at 50 - 60 days PP. The findings of our study suggested that day 60 PP served as a threshold for in vivo embryo production in Simmental cows.

Abstract Rabies is a viral disease affecting both land-dwelling and flying mammals. In India, dogs have been the primary source of rabies. This study aimed to investigate the role of wild animals in the transmission and persistence of the rabies virus and analyse the viral genome to understand its characteristics. In the present study, a brain sample from a fox (Vulpes vulpes) found dead in the free-ranging area of Mannamangalam forest station, Thrissur, Kerala state, India, was collected for rabies screening by fluorescent antibody staining and molecular techniques. Viral proteins and nucleic acids were detected and the samples were subsequently analyzed using next-generation sequencing for characterization. The genome analysis revealed that the virus belonged to the type I genotype of the arctic-like lineage. The phylogenetic analysis indicated that the fox virus strain shared close homology with all Indian isolates irrespective of the host species and was clustered in the same arctic-like lineage, denoting the distribution of a similar lineage and genotype across all regions of India. The deduced amino acid variations of nucleoprotein and glycoprotein genes also revealed a pattern of similarity amongst isolates of Indian origin and differed from isolates of other geographical locations and lineages. This study based on genome-wide surveillance could offer novel insights into the genetic makeup of currently circulating strains in the human-wildlife conflict and their continuing spread and persistence of rabies. 

Abstract The aim of this study was to investigate the presence of bovine herpesvirus type 1 and bovine parainfluenza virus type 3 (PI-3) in cattle lungs exhibiting pneumonia symptoms observed during macroscopic examination. Polymerase chain reaction, double immunohistochemistry, and double immunofluorescence methods were used for viral detection. For this purpose, during the summer-autumn period of 2023, a total of 84 lung samples from cattle aged 1 year and older, exhibiting macroscopic signs of pneumonia, were collected in Kayseri province, Türkiye. Pneumonia lesions were recorded according to the anatomical locations. In virus detection, PI-3 was identified at a rate of 8.33% using double immunohistochemistry/immunofluorescence staining methods, while this rate was found to be 6.72% in polymerase chain reaction. Bovine herpesvirus type 1 was not detected by any of the three methods. In the cattle lungs, 44 lesions (52.38%) were detected in the apical lobe, while seven lesions (8.33%) were detected in the middle lobe, eight lesions (9.52%) in the accessory lobe, and 25 lesions (29.76%) in the basal lobe. The PI-3 immunopositivity was observed in alveolar epithelial cells, as well as mononuclear cells in the interstitial and peri-vascular regions. This study is the first to examine bovine herpesvirus type 1 and PI-3 presence in bovine lungs using three distinct validation techniques (polymerase chain reaction, double immunohistochemistry, and double immunofluorescence).

Abstract Polycystic ovary syndrome (PCOS) is a complex endocrine disorder. Exercise is one of the interventions that leads to PCOS improvement. This study aimed to investigate the potential of different durations of high-intensity interval training (HIIT) or continuous training (CT) with/ without metformin (MET) in a rat model of PCOS. For this purpose, the rats were randomly placed in seven groups, including control, PCOS, PCOS + CT, PCOS + HIIT, PCOS + MET, PCOS + MET + CT, and PCOS + MET + HIIT. The PCOS was induced by a single intra-muscular injection of 2.00 mg kg-1 estradiol valerate. The concentration of lipid indices showed that serum cholesterol and low-density lipoprotein decreased, and high-density lipoprotein increased with 8 and 12 weeks of exercise training in PCOS + CT, PCOS + HIIT, PCOS + MET + CT, and PCOS + MET + HIIT groups compared to the PCOS group, but no significant difference was found between groups. Triglyceride level decreased in treated groups with 12 weeks of exercise compared to the PCOS group, and there was no significant difference between the treated groups. Finally, serum malondialdehyde level decreased in groups received exercise intervention alone or combined with MET compared to the PCOS group. These findings revealed that HIIT and CT with longer periods with/without MET can lead to improvement of lipid profile and malondialdehyde level of PCOS patients. 

Abstract Hyperuricemia, caused by impaired uric acid excretion, poses significant health risks. Urate oxidase (UOX) from Aspergillus flavus offers therapeutic potential by converting uric acid into soluble allantoin; however, its instability limits clinical applications. This study investigated the effects of osmolytes, including proline and glycine, on the kinetics and thermostability of recombinant A. flavus UOX. Following the expression of UOX coding sequence in Escherichia coli BL21, it was purified using Ni2+-NTA agarose affinity chromatography and confirmed by sodium dodecyl sulfate–polyacrylamide gel electrophoresis. The enzyme maintained its activity up to 35.00 ˚ C and lost its activity at higher temperatures as it lost 70.00 % of its activity after 60 min at 40.00 ˚ C, and the enzyme with proline and glycine additives maintained 73.00% and 30.00% of the activity, respectively. The inactivation rate constant of enzyme (k_in) was decreased in the presence of proline, indicating that the enzyme was more stable with proline, but glycine had no effect on kin. Half-life of enzyme was raised to 86 min in the presence of proline and the Michaelis constant (K_m) was decreased significantly by both osmolytes, as well. These results demonstrated that proline stabilized UOX by mitigating thermal denaturation, likely through preferential hydration and hydrophobic interactions, while glycine enhanced substrate binding. The stabilizing capacity of proline highlighted its utility for inclusion in biopharmaceutical formulations, offering a solution to the persistent challenge of UOX instability in therapeutic contexts. These findings yielded practical strategies for enhancing both structural integrity and catalytic performance of enzymes in pharmaceutical development.

Abstract Leishmaniasis comprises a spectrum of clinical manifestations caused by protozoan parasites of the genus Leishmania, order Trypanosomatida. Cutaneous leishmaniasis remains a significant zoonotic disease prevalent in tropical and subtropical regions, particularly in developing countries. Despite ongoing research, a definitive cure for this parasitic infection is still needed. This study explored the potential of activatable melittin (AM) as a selective treatment for cutaneous leishmaniasis caused by Leishmania major. The AM was designed using PepFold and ExPASy servers, incorporating a matrix metalloproteinase -2/9 cleavable linker to target L. major-infected macrophages selectively. To enhance drug delivery and reduce potential toxicity, AM was encapsulated within albumin nanoparticles and liposomes. The anti-leishmanial efficacy of these formulations was evaluated at AM concentrations ranging from 25.00 to 100 µg mL^-1 over 48 hr, with each experiment performed in 10 independent replicates (n = 10 per group). Statistical analysis using one-way ANOVA followed by Tukey's post-hoc test revealed a significant reduction in the average number of intracellular amastigotes per macrophage in the liposome-treated and albumin nanoparticle-treated groups (7.00 ± 1.50 amastigotes per macrophage) compared to the untreated infected control group (35.00 ± 3.20 amastigotes per macrophage). Treatment with 25.00 µg mL^-1 of AM encapsulated in non-toxic albumin nanoparticles and liposomes demonstrated the most promising anti-leishmanial effect, resulting in an approximately 80.00% reduction in intracellular L. major amastigotes (compared to control).

Abstract Reproductive failure is the major problem in modern pig production, significantly affecting economic productivity. This study investigated factors associated with reproductive failure in the hyper prolific pig farms, focusing on abortion, mummification, stillbirth and sudden increased sow death. Seven individual sows with reproductive failure clinical signs were surveyed to evaluate the infection rate with related pathogens. Results showed the highest frequency of Mycoplasma suis infections with reproductive problems along with blood samples being 100% positive, fetal mummification rate of 33.33% and stillbirth rate of 25.00%. Additionally, 28.57% of the total blood samples exhibited porcine circovirus type 3  infection, while other pathogens were not detected. The percentage of concurrent infections with M. suis and porcine circovirus type 3 in reproductive sow was 28.57%. The partial 16S rRNA Porcine Haemoplasma is a trivial name for haemotropic Mycoplasma spp., which can attach to the surface of red blood cells leading to deformity and anaemia in a wide range of mammalian animals including pigs. Haemoplasma sequences in this study was absolutely clustered into Haemominutum group. This study demonstrated the first case of acute reproductive disorder due to M. suis reported in imported hyper prolific sows.

Abstract Hamartomas are mostly benign tumor-like malformations characterized by a disorganized overgrowth of mature cells and tissues indigenous to the affected anatomical site. A 20-day-old male Kurdish foal was presented to the Veterinary Teaching Hospital of Urmia University, Urmia, Iran, with a history of hematochezia and tenesmus. Clinical examination identified a pedunculated tissue mass within the rectal lumen. The abnormal mass was surgically excised via polypectomy under local anesthesia. Tissue samples were obtained, fixed in 10.00% neutral buffered formalin, and processed routinely for histopathological examination. Histopathological evaluation of the resected tissue confirmed the diagnosis of a cystic polypoid rectal hamartoma characterized by normal rectal epithelium overlying multiple cystic, blood-filled vascular spaces. The foal recovered without complications and no recurrence was noted during a long-term follow-up exceeding 18 months. This report underscored that congenital rectal hamartoma, while rare, should be considered as a differential diagnosis in neonatal foals presenting with hematochezia or dyschezia. Prompt surgical intervention is curative and associated with a favorable prognosis.