Veterinary Research Forum

Abstract Mycobacterium avium subsp. paratuberculosis (MAP), the causative agent of Johne’s disease and a potential contributor to Crohn’s disease, presents a significant challenge due to its resistance to conventional antibiotics. This necessitates the development of innovative strategies for prevention and treatment. This study aimed to evaluate the anti-bacterial activity of pathogen-specific antibodies derived from chicken egg yolks (immunoglobulin Y [IgY]) and the postbiotics from lactic acid bacteria against MAP. Immunoglobulin Y antibodies were produced by immunizing hens with formalin-killed MAP strain antigens. The IgY was extracted and purified, and the anti-MAP titers were quantified by indirect enzyme-linked immunosorbent assay. The minimum inhibitory concentration of different concentrations of specific anti-MAP IgY and the mixture of postbiotics (from four different probiotic strains, including Lactobacillus reuteri, Lactobacillus rhamnosus, Lactobacillus acidophilus, and Pediococcus acidilactici) individually and in combination against MAP was determined at various time intervals. Anti-MAP IgY titers in egg yolks increased within 2 weeks of immunization, reaching peak levels at 6 weeks. Growth inhibition assays revealed that postbiotics concentration as low as 6.25 mg mL^-1 effectively inhibited MAP growth. Anti-MAP IgY demonstrated anti-bacterial activity with a minimum inhibitory concentration of 50.00 mg mL^-1, while the combined IgY-posbiotics treatment achieved MAP growth inhibition at a minimum inhibitory concentration of 3.125 mg mL^-1. The findings of the study suggest that combination therapy with specific IgY and postbiotics may be a promising preventive strategy for controlling MAP infections. Further in vivo studies are needed to elucidate the underlying mechanisms and optimize the application of this approach for broader use in veterinary and human medicine.

Abstract Testicular torsion-detorsion (TD) or ischemia/reperfusion causes reactive oxygen species over-production and has extensive destructive effects on testicular tissue. Following TD, this study evaluated the therapeutic effects of conditioned medium (CM) of fibroblast, macrophage and co-culture on testicular histomorphometric and ultrastructure. Adult rats were divided into 7 groups. Healthy control, Control sham, TD, and Dulbecco's Modified Eagle Medium groups. Experimental groups: Fibroblast, macrophage and co-culture. The supernatant was obtained from the separate cultures of fibroblasts and macrophages and co-culture were injected. All injections were made through rete testis. Thirty-five days after the operation, the testis was sampled for histomorphometric and ultrastructural studies. However, the ultrastructural study of testicular tissue also showed that extensive changes occurred in Sertoli cells' nucleus, nucleolus and mitochondria. This study showed that in fibroblast and slightly less in macrophage groups, there were good improvements in all histomorphometric and ultrastructure parameters similar to the healthy control group. It was also shown that the Dulbecco's Modified Eagle Medium group had slightly better recovery conditions than the TD group, but the co-culture group showed similar conditions to the TD. Overall, it could be concluded that the CM of fibroblast was very effective. In the groups receiving fibroblastic and macrophage CM, all parameters exhibited favorable improvement but in the fibroblast group, most of the parameters were similar to those in the healthy controls group. It was also shown that the CM of fibroblast-macrophage co-culture could not be improved on TD condition but made it worsened.

Abstract This study provided a comprehensive analysis of 145 urinary bladder stone cases diagnosed in dogs and cats over a 5-year period at a private veterinary hospital. The aim was to evaluate demographic patterns, urolith composition, and species-specific trends to enhance understanding and improve management of urolithiasis in companion animals. A retrospective study was conducted on clinical records, including patient demographics (species, breed, age, sex), urolith composition, and stone size. Chemical composition of uroliths was determined using qualitative chemical analysis. Statistical analyses were performed to identify significant associations between patient demographics and urolith composition. Of the 145 cases, 80 were dogs (55.17%) and 65 were cats (44.82%). In dogs, calcium oxalate stones predominated (65.00%) followed by struvite stones (30.00%) and mixed compositions (5.00%). Conversely, cats exhibited a higher prevalence of struvite stones (55.00%), with calcium oxalate accounting for 40.00% and mixed stones for 5.00%. Larger stones (10.00 - 20.00 mm) were more common in female dogs, while cats generally presented with fewer and smaller stones. No statistically significant temporal trend in urolith composition was observed for either species during the study period. Stone color distribution varied by species including cream and brown stones being most common in dogs and cats, respectively. This study established baseline epidemiological data on urolithiasis for this region, revealing significant species-specific differences in stone composition. The high prevalence of calcium oxalate in dogs and struvite in cats highlighted the need for tailored clinical management and provides a crucial benchmark for future regional and global comparative studies.

Abstract Leishmaniasis is a disease caused by Leishmania species and transmitted via sandflies. Current control strategies against reservoir hosts and vectors are not eco-friendly. Using harmaline (HA) from Peganum harmala, and meglumine antimoniate (MA) could be a promising therapy. The study aimed to explore the potential treatment outcomes and action mechanisms of HA and MA against Leishmania major stages by investigating their effectiveness through molecular docking, anti-leishmanial effects, safety assessment, and apoptotic profile evaluations. According to the molecular docking results, the protein-ligand interaction profiler identified that Bcl-2 interacts with HA mainly through hydrogen bonds, while Bax uses both hydrogen and hydrophobic interactions, indicating a stronger binding of HA to Bax compared to Bcl-2. The HA combined with MA (HA/MA) showed potent anti-leishmanial activity without toxicity. In vitro studies significantly demonstrated that HA inhibited the growth of promastigotes and amastigotes. The HA/MA was more effective in inhibiting parasite growth. Based on the study findings, HA and HA/MA mixture can be considered a viable treatment option for cutaneous Leishmaniasis.

Abstract To mitigate metabolic disorders and skeletal abnormalities associated with rapid growth in broilers, and to reduce reliance on antibiotics, strategies involving feed restriction and non-antibiotic dietary additives are increasingly explored. This study investigated the effects of various feeding methods and additives on weight gain, feed intake, feed conversion ratio, carcass characteristics, intestinal morphology and ileal digestibility in broiler chickens. The research involved 480 one-day-old Ross 308 male broiler chickens. A 2 × 5 factorial experiment, arranged in a completely randomized design, incorporating two feeding methods (without and white quantitative restriction) and four types of additives including thyme essential oil (300 g per ton), oregano essential oil (300 g per ton), equal weight ratio of thyme and oregano essential oil (300 g per ton) and probiotic (250 g per ton) along with control diets (without additives). Dietary supplementation with oregano essential oil significantly increased weight gain compared to the other additive groups. While the feeding method, additives and their interactions significantly influenced feed intake in chickens and the feed conversion ratio was primarily affected by the type of additive. Supplementation with any of the tested additives led to reduced total feed intake and improved final feed conversion ratio compared to the control group. Histological analysis revealed that chickens receiving oregano exhibited the highest villus height to crypt depth ratio. In conclusion, this research suggested that quantitative feed restriction, particularly when combined with specific dietary additives such as oregano essential oil could significantly improve broiler growth performance, feed efficiency and intestinal morphology, offering potential alternatives to conventional practices.

Abstract Crocetin (CRT) is one of the active chemical compounds of saffron and has many biological effects such as antioxidant property. The present study investigated the effects of CRT on crushed sciatic nerve function. Vitamin (Vit) C was used as an antioxidant drug. Thirty rats were divided into six groups including intact, sham, crush, CRT 7.50, CRT 30.00 and Vit C 100. Nine other rats with no surgery were scheduled in three groups to receive 7.50 and 30.00 mg kg^-1 CRT and 100 mg kg^-1 Vit C. In anesthetized rats, right sciatic nerve was crushed using a small hemostatic forceps. Sciatic functional index values on days five, 10, 15 and 20 after crush were accelerated, the severities of sciatic nerve degeneration and gastrocnemius muscle atrophy were ameliorated, and the increased malondialdehyde level and the decreased superoxide dismutase activity in the serum were restored by 20 consecutive days of oral administration of 30.00 mg kg^-1 CRT and 100 mg kg^-1 Vit C. No significant differences were observed between 30.00 mg kg^-1 and 100 mg kg^-1 Vit C. The groups that did not have surgery but received CRT (7.50 and 30.00 mg kg-1) and Vit C (100 mg kg-1) showed no behavioral, histopathological and biochemical alterations when compared to intact group. It was concluded that CRT and Vit C produced similar improving effects on crushed-injured sciatic nerve function. Inhibition of oxidative stress, enhancement of endogenous antioxidant activity might be involved in improving effects of CRT and Vit C.

Abstract Bovine viral diarrhea virus is a pestivirus of the Flaviviridae family including two biotypes, cytopathic (CP) and non-CP (NCP). This study aimed to evaluate DNA damage and apoptosis in Madin-Darby bovine kidney cells following infection with both biotypes. The MDBK monolayers were inoculated with a final dose of virus (1.00 × 10³ Tissue Culture Infectious Dose 50% mL-1) and incubated for 24 hr. DNA strand integrity was assessed using alkaline single-cell gel electrophoresis, and DNA damage was quantified through tail moment and olive tail moment indices (n = 3). Apoptosis was evaluated using annexin V–fluorescein isothiocyanate/propidium iodide flow cytometry to determine early and late apoptotic cell populations. Both biotypes significantly increased DNA fragmentation compared to the control group. The tail moment values were 15.89 ± 2.13 (control), 57.63 ± 16.20 (NCP), and 68.15 ± 9.93 (CP); while, olive tail moment values were 8.71 ± 1.01 (control), 29.35 ± 9.18 (NCP), and 35.14 ± 6.90 (CP). Apoptosis analysis showed a higher percentage of apoptotic cells in infected groups, with CP biotype of bovine viral diarrhea virus inducing the greatest early and late apoptotic responses, being consistent with its CP nature. Overall, both biotypes caused notable genomic injury and apoptosis in Madin-Darby bovine kidney cells, with CP producing the highest level of damage, confirming single-cell gel electrophoresis combined with apoptosis assays as sensitive tools for detecting virus-mediated genomic instability and supporting their potential application in breeding programs aimed at enhancing resistance to infectious diseases.

Abstract Selenium (Se) and gold (Au) nanoparticles (NPs) possess unique properties such as high absorption ability, low toxicity and minimal environmental persistence. These properties, combined with demonstrated anti-parasitic effects against certain parasites, suggest their potential as candidate for house fly control. This study aimed to evaluate the effectiveness of Se and AuNPs in controlling house flies. We investigated the insecticidal effect of Se and AuNPson Musca domestica using a dipping method. Several concentrations (20.00, 60.00, 100, 400, and 1,000 ppm) were prepared using deionized water. Fifteen larvae per concentration were tested, with each test repeated three times. The larvae were dipped in the NP solutions for 30, 60 and 90 sec in 10.00 mL of the tested concentrations. Distilled water was used for the control group. Larval mortality was recorded 24 hr post-treatment. No mortality was observed in house fly larvae 24 hr after exposure to different concentrations of Se and AuNPs, regardless of the incubation time. Thus, Se and AuNPs showed no insecticidal effect on house fly larvae. In conclusion, Se and AuNPs cannot be used to control house flies. Due to the issues associated with chemical control, it is crucial to search for alternative pesticides.