Veterinary Research Forum

Abstract Inflammatory bowel disease (IBD) encompasses a group of chronic inflammatory conditions that primarily impact the gastrointestinal system. While ulcerative colitis and Crohn's disease are the principal manifestations in humans, animals frequently exhibit lymphocytic-plasmacytic enteritis/colitis and eosinophilic enteritis/colitis. Growing evidence suggests a complex interplay among genetic predisposition, gut microbiota imbalance and abnormal immune responses to intestinal microbes in susceptible individuals. This intricate involvement results in remarkably similar clinical presentations across species. Patients often experience symptoms such as diarrhea, vomiting, weight loss and anemia. Extraintestinal manifestations including uveitis, skin rash and arthritis may also occur. Endoscopy and biopsy typically serve as the gold standard for confirming the diagnosis and differentiating it from other gastrointestinal disorders in humans and animals. The treatment approach generally focuses on managing disease activity through immunosuppressive medications such as glucocorticoids, administered at appropriate dosages. However, the precise cause of IBD remains a topic of ongoing research. With the emergence of additional treatment options like herbal compounds and fecal microbiota transplantation, which have demonstrated effectiveness in restoring gut health in IBD patients, there is optimism for novel therapeutic strategies. Ultimately, conclusion is that chronic gastrointestinal conditions like IBD are complex in both human and veterinary medicine. These diseases share numerous common pathophysiological features, yet, diagnostic and treatment challenges continue to exist.

Abstract The efficacy of eucalyptol on testicular ischemia-reperfusion (I/R) injury remains uncertain. This study aimed to investigate the protective effects of eucalyptol on I/R injury induced by testicular torsion/detorsion (T/D) in rats. A total of 32 rats were divided into four groups, including control, T/D, E100 (T/D + 100 mg kg^-1 eucalyptol administered 30 min prior to torsion), and E200 (T/D + 200 mg kg^-1 eucalyptol administered 30 min prior to torsion). Testicular T/D was induced in the left testis via a 720° clockwise torsion for 3 hr, followed by 3 hr of detorsion. Testicular tissues were harvested for histopathological, immunohistochemical including cleaved caspase-3, B-cell lymphoma 2 (Bcl-2)-associated X protein (Bax), Bcl-2, and proliferating cell nuclear antigen (PCNA), and biochemical parameter as glutathione peroxidase (GPx), reduced glutathione (rGSH), glucose-6-phosphate dehydrogenase (G6PD), vitamin C, and malondialdehyde (MDA) analyses. In the T/D group, significant reductions in GPx, rGSH, G6PD, and vitamin C levels were observed, alongside increased MDA levels compared to the control group. Immuno-histochemically, the T/D group exhibited increased expression of cleaved caspase-3 and Bax, along with decreased expression of Bcl-2 and PCNA, compared to the control group. Histologically, Johnsen and Cosentino scores were irregular in the T/D group. Pre-treatment with eucalyptol resulted in reduced MDA, cleaved caspase-3, and Bax levels, while GPx, rGSH, vitamin C, G6PD, PCNA, and Bcl-2 levels increased. Additionally, improvements in Cosentino and Johnsen scores and histopathological damage were observed. These findings suggest that eucalyptol may exert a protective effect against I/R injury caused by testicular T/D, likely due to its anti-oxidant and anti-apoptotic properties.

Abstract Foot-and-mouth disease virus (FMDV) is a highly transmissible pathogen causing severe economic losses in the global livestock sector. Frequent outbreaks of FMDV type-O in Pakistan highlight the need for continuous genomic and antigenic surveillance to track its evolution. This study aimed to isolate and molecularly characterize FMDV type-O from an outbreak in the Punjab province of Pakistan using in vitro cell culture techniques. Samples were processed for viral isolation on a susceptible cell line, followed by RNA extraction. The VP1 gene, pivotal for antigenicity and immunogenicity, was amplified using a one-step polymerase chain reaction protocol. Purified amplicons underwent sequencing, and the nucleotide sequences were translated into amino acid sequences for further analysis. Protein three-dimensional modeling and in silico comparison were performed against the vaccinal seed strain PanAsia-2. The VP1 sequence analysis revealed notable genetic variability among the isolates, indicating adaptive evolution. Structural and antigenic modeling uncovered key differences between the field isolates and vaccinal strain, suggesting potential antigenic drift, which could undermine vaccine performance. The study underscores the dynamic evolution of FMDV type-O in Pakistan and the critical importance of ongoing genomic monitoring to refine vaccine strategies and enhance outbreak control.

Abstract This study examined the effects of alpha-ketoglutaric acid (AKG; 2.00, 4.00, and 8.00 mM L^-1) and folic acid (FA; 50.00 nM L^-1) on bull sperm cells under ammonia stress induced by 175 µM L^-1 ammonium chloride. Sperm parameters including kinematic motility, survival rate, membrane integrity, DNA integrity, superoxide dismutase (SOD), glutathione peroxidase (GPx), catalase (CAT), total antioxidant capacity (TAC) activities, and apoptosis were assessed in the groups. Sperm motility indices, survival rate, plasma membrane integrity, SOD, CAT, TAC, and GPx enzymes activity, DNA damage, and apoptosis in the treated groups were significantly differed from those in the control groups. The AKG concentrations of 4.00 and 8.00 mM L^-1 co-supplemented with 50.00 nM L^-1 FA mitigated the negative effects of ammonia on sperm cells. This study indicated that supplementation with AKG and FA at the desired concentrations counteracted the adverse effects of ammonia toxicity that preceded clinical signs. Further studies are needed to evaluate the fertility of these sperms, either in vitro or in vivo.

Abstract Chemotherapy agents impose harmful side effects beside their therapeutic properties. This study aimed to uncover the ameliorative effects of thiamine (VitB1) on sperm characteristics and in vitro fertilizing ability in methotrexate (MTX) exposed mice. For this purpose, 36 apparently healthy adult male mice were randomly divided into six groups (n = 6), including control (0.10 mL normal saline per day; intraperitoneally [IP]), sham (10.00 mg kg^-1 per week MTX; IP), positive control group (100 mg kg^-1 per day VitB1; IP), and three experimental groups (25.00, 50.00, and 100 mg kg^-1 per day; IP VitB1 plus MTX, respectively; IP). Then, sperm parameters and the in vitro fertilizing potentials were assessed. The sperm count, motility, and viability in the sham group showed a significant decrease in comparison with the VitB1-treated and control groups. In the VitB1 100 mg kg^-1 + MTX group, there was a significant increase in sperm count in comparison with the sham group. The sperms with DNA damages and immature sperms in the sham group significantly increased in comparison with the positive control and control groups. In vitro fertilization and embryonic development in the sham group were decreased in comparison with both the control and VitB1 groups. In VitB1 50.00 mg kg^-1 and 100 mg kg^-1 + MTX groups, there was an increase in the fertilization rate, along with embryonic development promotion. According to the results of this study, thiamine ameliorates toxic effects of MTX on sperm traits and in vitro fertilizing potentials by constraining oxidative stress.

Abstract Bovine respiratory disease (BRD) is a significant disease in the cattle industry worldwide. The interaction between environmental factors, hosts, livestock management, and viral and bacterial pathogens causes this disease. Viruses are crucial in the initiation and progression of BRD. This study was the first to investigate the prevalence of BRD viruses using the reverse transcription polymerase chain reaction method in nasal and eye conjunctival swabs and blood samples of 115 BRD calves in the central desert of Iran. At least one investigated virus was detected in 44 animals (38.26%). The detection rates of bovine viral diarrhea virus, bovine coronavirus, bovine adenovirus, bovine respiratory syncytial virus, bovine herpes virus-1, and bovine para influenza virus-3 were 20.00, 14.78, 5.21, 0.86, 0.00, and 0.00%, respectively. Three animals (2.60%) had a simultaneous infection with two viruses. Detection of bovine viral diarrhea virus, bovine coronavirus, and bovine adenovirus was correlated. The virus infection rates were 31.81 and 44.66% in five sampled cities. The virus detection rate in infected animals was related to the nose (26 animals; 50.09%), nose and eyes (seven animals; 15.90%), eyes (seven animals; 15.90%), nose, eyes, and blood (three animals; 6.81%), and blood (one animal; 2.27%) samples. The virus detection rate in different samples was in separate clusters. Monitoring and controlling the circulation of bovine viral diarrhea virus and bovine coronavirus in the central desert of Iran is vital due to the high detection rate. Our results highlight the necessity of investigating other viruses and bacterial agents related to the BRD in the study area.

Abstract Today, a combination of immunological and bioinformatics tools has become common for vaccine design, making vaccine production affordable. Considering the importance of recombinant protein purification for vaccine production in a cost-effective way, our study aimed to in silico design a fusion protein vaccine against egg drop syndrome virus (EDSV) with a higher isoelectric point for more affordable purification. The in silico design of fusion protein, including egg white lysozyme and fiber protein as an antigen from egg drop syndrome virus, was performed. In addition to isoelectric point changing, lysozyme probably helps the antigenicity by increasing the size of the antigen. Also, lysozyme can act as a preservative. The physicochemical characteristics, stability, secondary and tertiary structure, epitope prediction, antigenicity, and mRNA structure were analyzed using computational and bioinformatics tools. The results showed that the isoelectric point of the gene construct was 8.87, which can be purified by ion exchange chromatography. Validation of the Ramachandran plot showed that the predicted model was accurate and suitable. The tertiary structure of the fusion protein was modeled as well, and its trimer structure, being required for immunogenicity, was preserved. The antigenicity of the target construct was also suitable. Protein was stable and hydrophilic based on aliphatic index and grand average of hydropathicity, which can be a good candidate for a vaccine. After experimental studies, this fusion protein may be used as a vaccine against EDSV.

Abstract Male and female broiler chickens differ in performance and this will cause unwanted experimental errors in research. For this reason, single-sex or mixed-sex broilers are used in most studies. This study aimed to assess the performance differences between groups of chickens with varying male/female ratios to determine how sex ratio can affect performance criteria. Birds (N = 550) were separated by sex and placed in 11 groups (pens) according to the male/female ratios, including group 1 (10 males + 0 female), group 2 (9 males + 1 female), group 3 (8 males + 2 females), group 4 (7 males + 3 females), group 5 (6 males + 4 females), group 6 (5 males + 5 females), group 7 (4 males + 6 females), group 8 (3 males + 7 females), group 9 (2 males + 8 females), group 10 (1 male + 9 females), and group 11 (0 male + 10 females). The results showed that male broiler chickens had higher feed intake and body weight gain than female broiler chickens, but the feed conversion ratio was not affected by gender. The digestibility of phosphorus, bone strength, bone density, bone calcium and phosphorus, pH, and redness and water holding capacity of meat were higher in male broilers. The dripping loss was higher in female broilers. This study showed that male and female broiler chickens differed in most of the parameters examined in the research, and the use of separate breeding affected the research results.