Veterinary Research Forum

Abstract Equine glandular gastric disease (EGGD) has limited prevalence studies and scarce understanding of the predisposing factors and pathophysiological mechanisms involved. The objectives of this study were to determine and compare the post-mortem prevalence of EGGD and other gastric lesions in Colombian equids, specifically horses, donkeys, and mules. The study utilized a sample of 300 equids stomachs, with 100 specimens from each group, collected immediately after slaughter. The post-mortem examination included a thorough inspection of the ventral fundus, adjacent area of the margo plicatus, and pyloric antrum of the glandular mucosa, carefully identifying any lesions or abnormalities. The results showed that the overall macroscopic presence of EGGD in the equid population was 68.30% on average, with specific rates of 45.00% in horses, 77.00% in donkeys, and 83.00% in mules, with lesions predominantly affecting the fundus and pyloric antrum regions. Inflammatory processes were observed to be similar between donkeys and horses, while mules exhibited distinct inflammatory patterns in the glandular mucosa. Additional gastric findings, such as gastritis (85.30%), gastric impaction (10.00%), and parasitic infections (13.30%), were also documented during the study. The findings suggested that EGGD was prevalent across all three groups of equids. Although the inflammatory conditions and their distribution were similar in horses and donkeys, differences in the location, frequency, and severity of mucosal lesions were observed among the three species.

Abstract This study aimed to detect Coxiella burnetii, Chlamydia abortus, and Brucella species in the abomasal contents of aborted ruminant fetuses from the Central Anatolia region of Türkiye using PCR between 2020 and 2023. The abomasal contents of a total of 97 aborted fetuses from cattle, sheep, and goats with a history of abortion, collected between the years 2020 and 2023, were tested in this study. As a result of PCR analysis of 97 abomasal contents, four (4.10%; 95.00% confidence interval [CI]: 1.33 - 10.82) of them were C. abortus, including three sheep and one goat. Two (2.10%; 95.00% CI: 0.36 - 7.96) of them were C. burnetii, including one sheep and one cow. A total of 60 (61.90%; 95.00% CI: 51.40 - 71.37) samples from 47 cattle, nine sheep, and four goats were determined by Brucella genus-specific PCR. Following multiplex PCR analysis of the positive Brucella spp. samples, 39 (65.00%; 95.00% CI: 51.52 - 76.55) samples were identified as B. abortus, including two sheep, one goat, and 36 cattle. Additionally, 19 (31.70%; 95.00% CI: 20.60 - 45.09) isolates were identified as Brucella melitensis, including five sheep, two goats, and 12 cattle. In two sheep samples, both B. melitensis and C. abortus were identified from the same animals. In conclusion, Brucella spp. were the predominant abortion-causing pathogens, with C. abortus also contributing significantly. Effective control strategies under the One Health approach are essential to prevent the uncontrolled spread and inter-species transmission of these zoonotic agents in the region and country.

Abstract Effective reproductive management in cattle, such as cows and buffaloes, requires early and accurate pregnancy detection. Early identification of pregnancy enables farmers to promptly identify non-pregnant animals for treatment and/or rebreeding, thereby reducing the calving interval. This study aimed to standardize the expression of the CCL8 and CXCL10 genes as markers for early pregnancy detection in Murrah buffaloes. Blood samples were collected on the 16th day post-artificial insemination for gene expression analysis and on days zero, seven, 14, and 21 post-artificial insemination for progesterone concentration measurement. Buffaloes were categorized as pregnant (n = 6) or non-pregnant (n = 6) based on the resumption of estrus. Gene expression levels in peripheral blood leukocytes were analyzed using Quantitative Reverse Transcription Polymerase Chain Reaction (qRT-PCR) with SYBR green dye. Amplicons of CCL8, CXCL10, and GAPDH genes were measured 108, 117, and 158 bp, respectively. Results showed that CCL8 mRNA expression in pregnant buffaloes was 5.13 and 12.21 times higher compared to non-pregnant buffaloes, while CXCL10 mRNA expression was 4.19 and 22.17 times higher. These findings indicated significantly elevated CCL8 and CXCL10 mRNA expression levels in peripheral blood leukocytes of pregnant buffaloes on the 16th day. Progesterone levels in the pregnant group were increased significantly from day zero to day 21, while no significant differences were observed between groups on days zero, seven and 14. Pregnancy was further confirmed via per-rectal examination on the 45th day post-artificial insemination Therefore, CCL8 and CXCL10 gene expression profiling on the 16th day could serve as reliable early pregnancy markers in Murrah buffaloes.

Abstract Cadmium (Cd) is a highly toxic environmental pollutant known to cause severe damage to the male reproductive system. This study aimed to investigate the protective effects of bee bread (BB), a natural product with anti-oxidant, anti-apoptotic, and anti-inflammatory properties, against Cd-induced testicular toxicity in male Wistar rats. A total number of 32 rats were divided into four groups, including control, BB (0.50 g kg^-1), Cd (5.00 mg kg^-1), and Cd + BB (5.00 mg kg^-1 and 0.50 g kg^-1, respectively) groups. Administrations via oral gavage were performed for 4 weeks. Semen analysis revealed significant reductions in sperm motility and density along with increases in abnormal and dead sperm ratios in the Cd and Cd + BB groups compared to controls. Histopathological examination showed severe degeneration and desquamation of germ cells, tubular atrophy, and a decrease in spermatozoa in the Cd-treated groups. Polymerase chain reaction analysis indicated up-regulation of apoptotic markers (caspase-3, -8, and -9) and oxidative stress enzymes (catalase and superoxide dismutase) in the Cd group, signifying disrupted testicular function. The BB administration partially mitigated Cd-induced damage as evidenced by less severe histopathological changes and moderated gene expression alterations. However, the protective effects of BB were not sufficient to completely counteract the toxic impact of Cd. The present study concluded that while BB had potential in reducing Cd-induced testicular toxicity, its protective efficacy was limited, warranting further research to explore its therapeutic potential in combination with other protective agents.

Abstract Thymoquinone (TQ), the bioactive compound found in black seed, possesses beneficial properties. In the present study, the effects of oral administration of TQ were investigated on acute corneal and orofacial pains in rats. To clarify the central mechanism of action, muscarinic cholinergic, cannabinergic 1 (CB1) and 5-hydroxytryptamine receptor antagonists were delivered into the 4^th ventricle of the brain after oral administration of TQ. Acute corneal and orofacial pains were induced by dropping of hypertonic saline (50.00 µL; 5.00 M) on the corneal surface and subcutaneous injection of capsaicin (1.50 µg; 20.00 µL) in the vibrissal pad, respectively. The eye wiping number and face rubbing duration were recorded as corneal and orofacial pains behavioral responses, respectively. Locomotor activity was measured using an open-field test. The TQ (5.00 mg kg^-1) had no effects, while it reduced pain responses at 10.00, 20.00, and 40.00 mg kg^-1. Intracerebro-ventricular injections of naloxone (an antagonist of opioid receptors), (N-(Piperidin-1-yl)-5-(4-iodophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide (an antagonist of CB1 receptors), atropine (an antagonist of muscarinic cholinergic receptors), and ondansetron (an antagonist of 5-hydroxytryptamine 3 receptors) at a similar dose of 10.00 µg kg^-1 inhibited corneal and orofacial pains suppression caused by 40.00 mg kg^-1 TQ. The tested drugs did not affect locomotor activity. It is concluded that TQ caused analgesia in the acute corneal and orofacial pains. Central opioid, cholinergic muscarinic, CB1, and 5-hydroxytryptamine 3 receptors might be involved in the anti-nociceptive effects of TQ.

Abstract Obese and overeating children are at risk of obesity and its complications in adulthood. Research on childhood obesity encounters numerous challenges in mammals. Broiler chicks are a suitable animal model for studying childhood obesity. Genetically, broiler chicks exhibit high growth rates. They are hyperphagic, hyperglycemic, and capable of accumulating abdominal fat, and their diet can be managed from birth. Soybean oil, which is rich in omega-6 (n-6) polyunsaturated fatty acid linoleic acid and low in omega-3 (n-3) polyunsaturated fatty acid (n-6 : n-3 ratio = 7.50), is widely utilized in human nutrition. However, conflicting findings have been reported regarding the efficacy of this oil in humans and rodents. Effects of a soybean oil-enriched diet (4.00% total fat as a control vs. 11.00% total fat as a treatment) on metabolic disorders in broiler chicks were evaluated from hatching to 36 days of age. Results showed no changes in food intake, body weight, appetite-regulating neuro-peptide mRNA levels, blood triglycerides, or hematological parameters. In contrast, the relative abdominal fat, blood cholesterol, aortic wall thickness, intima layer, and area of fat cells increased significantly in treatment group compared to control group. Signs of liver fat infiltration (steatosis) and changes in the aortic intima layer, including increased distance between elastin fibers, were observed. In conclusion, in middle-term-fed broiler chicks, a model for childhood obesity using soybean oil high in omega-6 polyunsaturated fatty acids leads to early atherosclerosis, fatty liver, adipose dysfunction, and hypercholesterolemia, without impacting body weight or food intake.

Abstract Biofilm formation is a key virulence factor in Staphylococcus aureus, contributing to bacterial persistence, antimicrobial resistance, and chronic infections. This study aimed to investigate the presence of biofilm-associated genes (fib, fnbA, fnbB, clfA, and clfB) in S. aureus isolates from dogs in Ilam, Iran. From December 2022 to September 2023, 250 swab samples were collected from nasal, oral, and rectal sites of dogs, yielding 81 S. aureus isolates confirmed by PCR amplification of the nuc gene. The prevalence of biofilm-associated genes varied, with clfA, clfB, and fnbA detected in 98.80% of isolates, fib in 63.00%, and fnbB in 16.00%. Notably, fnbA, clfA, and clfB were present in all rectal isolates, while fnbB was absent in this group. The findings highlighted the widespread presence of biofilm-related genes in S. aureus from dogs, suggesting their potential role in colonization and zoonotic transmission. The high prevalence of adhesion-associated genes underscored the need for monitoring biofilm-forming S. aureus in companion animals to mitigate antimicrobial resistance and public health risks.

Abstract Dicrocoeliasis is a globally significant condition impacting both economic and public health. The lack of effective vaccines and emergence of drug-resistant flukes have prompted research into alternative treatments. Metallic nanoparticles have recently been studied for their potential as anthelmintic agents. This research examined the in vitro anthelmintic activity of copper oxide (CuO-NPs) and zinc oxide nanoparticles (ZnO-NPs) against Dicrocoelium dendriticum. Using adult motility inhibition tests and oxidative stress biomarkers, including glutathione peroxidase , glutathione S-transferase , superoxide dismutase , and malondialdehyde , this study evaluated the effects of CuO-NPs and ZnO-NPs. Flukes were treated with various concentrations of nanoparticles (1.00, 4.00, 8.00, 12.00, and 16.00 ppm) for 24 hr. The CuO-NPs and ZnO-NPs demonstrated concentration- and time-dependent anthelmintic activity. Higher concentrations (12.00 and 16.00 ppm of CuO-NPs, and 16.00 ppm of ZnO-NPs) significantly inhibited worm motility compared to the controls. The nanoparticles induced oxidative stress in the flukes, with decreased superoxide dismutase, glutathione S-transferase, and glutathione peroxidase activities and increased malondialdehyde levels. Based on these findings, CuO-NPs and ZnO-NPs exhibit potential as therapeutic agents for controlling and treating D. dendriticum. However, further studies are necessary to assess their safety and efficacy in vivo for managing parasitic infections.

Abstract Numerous studies have explored the molecular epidemiology of H9N2 viruses in Iran; however, continuous monitoring remains vital for timely interventions to mitigate potential damage. This study examined the molecular characteristics and evolutionary features of Iranian H9N2 viruses by sequencing the complete genomes of two viruses, Marand and Baneh, isolated in 1998 and 2022, respectively, alongside other Iranian strains from GenBank. All Iranian viruses were identified as low-pathogenic avian influenza viruses, as evidenced by the presence of the di-basic motif K/RSSR cleavage site. Notably, all Iranian viruses isolated from 2009 onward had an L at position 216 in the hemagglutinin receptor binding site, whereas earlier viruses exhibited a Q/L at the same position, an essential mutation that enhances replication in mammalian cells. The molecular evolutionary rates for the Iranian hemagglutinin (HA) and neuraminidase (NA) genes were estimated at 4.50 × 10^-3 and 3.60 × 10^-3 substitutions per site per year, respectively. Error-prone replication of H9N2 viruses has resulted in the continuous evolution of Iranian strains over two decades, characterized by three phases of population growth. Maximum likelihood phylogenetic analysis revealed that the HA and NA genes of H9N2 viruses from domestic chickens belonged to the G1 sublineage. Additionally, the internal genes of some viruses displayed evidence of reassortment with other subtypes, indicating potential gene exchange with other viruses. These findings underscored the importance of ongoing surveillance of H9N2 viruses in both domestic and wild bird populations, given the human-like receptor-binding preference and the possibility of genetic reassortment with various viral subtypes.

Abstract The effects of substituting whole egg yolk (WEY) in a tris-citrate-based extender with two derived fractions, including a buffer-soluble fraction (BSF) and an ammonium sulfate insoluble yolk fraction, on the freezability of bull sperm were investigated. The BSF and ammonium sulfate insoluble yolk fraction levels of egg yolk in the respective diluents were consistent with their extracted values from an equivalent volume (20.00%) of egg yolk in the control diluent. The extenders were then enriched with 0.00, 5.00, and 10.00% (v/v) of pasteurized skim milk. Semen samples were collected weekly over five consecutive weeks from six adult bulls, and the standard ejaculates were pooled. The pooled semen was subsequently divided into seven experimental extenders and frozen in 0.50 mL French straws. Various sperm quality parameters, including kinematics, acrosome integrity, capacitation, and DNA fragmentation, were evaluated post-thawing. Results indicated that sperm kinematics and the percentage of acrosome-reacted sperm in experimental extenders were not significantly different from those in the control group. The milk-free ammonium sulfate insoluble yolk fraction extender exhibited higher percentages of capacitated sperms, but lower percentages of spermatozoa with intact DNA compared to the WEY extender sperm. The addition of 10.00% milk into the BSF diluent resulted in a significant increase in the proportion of sperm with intact DNA and a notable decrease in the percentage of sperm with partially fragmented DNA compared to the control. In conclusion, a BSF extender enriched with 10.00% cow's skimmed milk is recommended as a substitute for WEY in the cryopreservation of bull semen.