Veterinary Research Forum

Abstract The amygdala is the nucleus of the brain that is largely responsible for perceiving danger and plays a role in emotion, behavior, control and learning. A small amygdala has been associated to aggression. Horned ewes are expected to be more aggressive and have a smaller amygdala. Both horned and hornless ewes exhibit intraspecific head-butting behavior and both species are at risk for traumatic brain injury. The aim of this study was to investigate the neuronal density, glial cells and blood-brain barrier (BBB) of the amygdala in horned and hornless ewes. Four horned and six hornless ewe heads (age: 16.00 ± 4.00 months) were obtained from the abattoir. The brains were carefully removed and preserved in 10.00% formalin for 5 days. Bilateral amygdalae were sectioned. The samples were stained with Hematoxylin and Eosin, immunohistochemical (glial fibrillary acidic protein) and Terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick end labeling methods, and the histological structures of the amygdala were examined by light microscopy. The Mann-Whitney U test was used to analyze the data. Neuronal density was estimated to be 143,230 ± 12,540 per mm3 in horned and 152,230 ± 18,430 per mm3 in hornless ewes. Horned subjects had reduced numbers of neurons, damaged BBB and localized inflammatory areas. More apoptotic neurons were observed in horned ewes. Further studies are needed to determine whether these differences in neuronal density, glial cells, and BBB are acquired (due to trauma) or congenital. The results of this study might need further similar studies to be conducted in the future.

Abstract In vitro maturation is a critical step in in vitro fertilization, significantly impacting oocyte quality and subsequent embryonic development. This study examines how insulin-like growth factor (IGF)-1 supplementation in the maturation medium affects mitogen-activated protein kinase and maturation promoting factor expressions, as well as the amount of mitochondrial DNA copies in Kacang goat oocytes. Oocytes were collected from Kacang goat ovaries and matured in vitro with varying IGF-1 concentrations, including 0.00 (control), 50.00, 100, and 150 ng mL^-1. Immuno-cytochemistry was used to assess mitogen-activated protein kinase and maturation promoting factor expressions, while quantitative polymerase chain reaction quantified mitochondrial DNA copy number. Results showed that the 100 ng mL^-1 dose of IGF-1 group had substantially increased mitogen-activated protein kinase and maturation promoting factor expressions and also mitochondrial DNA copy numbers compared to the other groups. These findings suggest that IGF-1 supplementation with 100 ng mL^-1 dose optimally enhances oocyte maturation by activating key signaling pathways and promoting mitochondrial replication. In conclusion, IGF-1 supplementation at 100 ng mL^-1 is recommended to improve oocyte quality in Kacang goats, potentially enhancing in vitro fertilization outcomes.

Abstract Staphylococcus aureus is one of food poisoning bacteria. This study assessed the prevalence, virulence factors, and antimicrobial susceptibility profiles of S. aureus isolated from frozen fish and shrimp in Egypt. Ninety samples from frozen fish fillets and shrimps (45 for each) were examined for S. aureus prevalence using VITEK 2 compact system, followed by molecular confirmation by nuc gene, virulence characterization, and its resistance genes. The overall prevalence rate of S. aureus was 14.44% (13/90). Fish fillet had the highest mean total S. aureus count (9.50 ± 3.50 × 10⁵ colony forming unit g^-1), followed by shrimp (7.50 ± 3.30 × 10⁵ colony-forming unit g^-1), with a non-significant difference among fish fillet and shrimp. All confirmed S. aureus isolates were lecithinase producers and showed β-hemolysis, and coagulase positive, and confirmed molecularly coa gene positive. All isolates were resistant to ampicillin (100%), both erythromycin and clindamycin (69.23%), and tetracycline (61.53%), followed by vancomycin (46.15%). However, all isolates were sensitive to linezolid, tigecycline (84.70%), and trimethoprim/sulfamethoxazole (61.53%). Twelve (92.30%) phenotypically cefoxitin and oxacillin-resistant and molecularly mecA recovered S. aureus isolates were confirmed as methicillin-resistant S. aureus, while based on vancomycin-resistant pheno-typically, and molecularly vanA recovered S. aureus isolates were confirmed as vancomycin-resistant S. aureus. The emergence of multidrug-resistant methicillin-resistant S. aureus and vancomycin-resistant S. aureus in frozen fish fillets and shrimp indicates public health hazards, so there is a need for food safety measures alongside reliable detection methods of resistant bacteria along the food chain.

Abstract Anti-oxidants are vital for protecting sperm and can mitigate the negative effects associated with cryopreservation. This study was conducted to evaluate the protective effects of adding anti-oxidants (vitamins and amino acids) to extender in sex-reversed rainbow trout (Oncorhynchus mykiss) sperm. The collected sperm was diluted at the ratio of 1 : 5 by the extenders supplemented with different anti-oxidants including 1.00 mM of ascorbic acid  and L-tryptophan, 2.00 mM of cysteine and α-tocopherol, and their combination. After dilution, the semen was aspirated into 0.50 mL straws, and the straws were placed on the tray, frozen for 10 min, and plunged into liquid nitrogen. Straws were thawed in a 30.00 ˚C water bath for 15 sec. The Sperm Class Analyzer System was used to evaluate sperm kinematics. The activity of anti-oxidant enzymes and lipid peroxidation were determined as oxidative stress indices. Our data indicated that the incorporation of anti-oxidants and amino acids increased sperm motility duration. The elevated activity of glutathione peroxidase, superoxide dismutase, and catalase in post-thaw samples indicates that the anti-oxidant system in sex-reversed rainbow trout sperm likely plays a crucial role in protecting membrane compounds from oxidation. In conclusion, the combination of 1.00 mM L-tryptophan and ascorbic acid to the extender media caused a prolonging effect in sperm motility after thawing and they have the potential to serve as effective agents for improving sperm cryosurvival.

Abstract Goose parvovirus causes major economic losses in the waterfowl industry due to the high mortality. Therefore, it is essential to establish protection/control objectives in the fight against the disease. The genome of goose parvovirus consists of three structural proteins, including VP1, VP2, and VP3. The VP2 is a candidate antigen in developing vaccines and diagnostic kits. This study aimed to produce the VP2 protein from a local goose parvovirus strain that causes serious infections in geese in Türkiye using the baculovirus expression vector system. To achieve this, the VP2 gene was first amplified by polymerase chain reaction, followed by purification and insertion into the pENTR™/TEV/D-TOPO™ entry vector. Then, the target gene in the pENTR™/TEV/D-TOPO™ vector was transferred to linear N-Term BaculoDirect™ DNA through LR recombination (site-specific recombination between attL and attR sites). The construct was transfected into Spodoptera frugiperda cells. To verify the production of baculoviral virions, a band of approximately 600 bp in length was obtained as a result of polymerase chain reaction amplification using external primer sets for both the VP2 gene and expression vector. The obtained band was purified and sequenced for confirmation. In addition, to confirm the production of the recombinant protein, western blot analysis was conducted utilizing the V5 epitope located at the N-terminus of the expressed protein, resulting in the detection of a ~65.00 kDa band corresponding to the VP2 gene. To detect protein expression in S. frugiperda cells infected with the recombinant baculovirus, immunofluorescence analysis was performed using the same epitope. 

Abstract Use of various nanoparticles and vitamins to strengthen the immune system of fish to improve growth and biochemical indice, and to mitigate the harmful effects of free radicals through enhanced antioxidant enzyme activity has become the subject of numerous studies. In this study, fish with an average weight of 55.20 ± 7.90 g, after a two-week acclimation period, were randomly distributed into 18 polyethylene 300-L tanks, with 25 fish per tank. The experiment consisted of six treatments with three replicates: Control, vitamin (Vit) C100, nano-selenium (N-Se)0.40, Vit C100 + N-Se0.10, Vit C200 + N-Se0.20, and Vit C400 + N-Se0.40. Blood samples were collected post-treatment for analysis of antioxidant enzyme activities, biochemical and hematological indices as well as post-handling stress. Results indicated significant differences in antioxidant enzyme activities among treatments. The highest catalase activity was observed in Vit C100 and Vit C100 + N-Se0.10 groups. Glutathione peroxidase activity was significantly higher in the control and Vit C200 + N-Se0.20 groups. Malondialdehyde was the highest in Vit C100 + N-Se0.10 group. Superoxide dismutase activity peaked in Vit C200 + N-Se0.20 group. Biochemical indices showed significant differences among treatments except for high-density lipoprotein. Hemoglobin and red blood cell counts were significantly different. The Vit C100 + N-Se0.10 group showed the highest serum cortisol and glucose levels post-handling stress. Overall, dietary N-Se and Vit C supplementation improved antioxidant, biochemical, hematological indices and stress resistance in rainbow trout.

Abstract In recent years, liver transplantation has emerged as the standard therapy for several liver disorders. Throughout the procedure, the transplanted liver tissue is subjected to varying degrees of ischemia-reperfusion (IR) damage. Consequently, there has been a long-standing pursuit of substances that can alleviate the harm caused by IR. In our investigation, we employed dapagliflozin as a potential therapeutic agent. Eighteen Wistar rats were divided into three groups (n = 6), including treatment, IR, and control that did not undergo surgical intervention. Two days prior to surgery, the treatment group received dapagliflozin at a dosage of 10.00 mg kg^-1 orally. During surgery, liver ischemia was induced for 1 hr, followed by a 24-hr reperfusion period. The IR group exhibited elevated levels of alanine transaminase, aspartate transaminase, alkaline phosphatase, bilirubin, lactate dehydrogenase, and malondialdehyde compared to the control group. In contrast, the treatment group showed levels of these factors that were closer to those of the control group. While total protein, albumin, and total anti-oxidant capacity decreased in the IR group, this decline was less significant in the treatment group. Analysis of oxidative stress in liver tissue revealed that the treatment group had increased anti-oxidant capacity, and exhibited less oxidative stress compared to the IR group. Furthermore, dapagliflozin was found to reduce the degree of liver edema, necrosis, and vascular hyperemia following IR. Overall, dapagliflozin demonstrates the potential to lessen liver damage, enhance liver tissue regeneration, and mitigate the consequences associated with liver impairment.

Abstract Diabetes is a metabolic disorder characterized by hyperglycemia due to the defects in insulin secretion and function, or both. Glabridin (GBD) is one of the natural anti-oxidants used for infertility treatment. This study was planned to evaluate the effects of GBD on testicular oxidative status, sperm characteristics, and early embryo development in diabetic mature mice. Forty mature male mice were allotted to five equal groups, including control group received no treatment, diabetic group received intraperitoneal streptozotocin (50.00 mg kg^-1), and three experimental groups receiving 12.50, 50.00, and 200 mg kg^-1 GBD by gavage daily for 30 days, respectively. Serum levels of testosterone, sperm parameters, and testicular malondialdehyde, total anti-oxidant capacity, and catalase levels, as well as pre-implantation embryo development were determined. The diabetic group exhibited significantly reduced sperm motility, viability, and count, testosterone level, and testicular total anti-oxidant capacity and catalase levels, and increased testicular malondialdehyde level, and DNA-damaged and immature sperms along with poor in vitro fertilization outcomes compared to the control group. In contrast, the GBD administration, particularly at the highest dose, caused a pronounced improvement in the above-noted parameters. These findings suggest that GBD may play a role in impeding diabetes-induced male reproductive complications through oxidative stress repression, and sperms and early embryos protection.

Abstract Rotavirus is one of the major causes of diarrhea in different animal species and has a bad economic impact due to the losses in neonates and productivity. To investigate the occurrence of this infection in bovine calves, three localities in Khartoum State, Sudan, were selected. A total of 200 fecal samples were collected from diarrheic calves; 100 from Khartoum and 50 from each of Khartoum Bahari and Omdurman provinces. Collected samples were screened for a group A rotavirus antigen using enzyme-linked immunosorbent assay (ELISA). Positive results were seen in 40.00% of samples; the highest prevalence of 42.00% was found in samples from Khartoum province. Five ELISA-positive samples were examined under electron microscope, and characteristic wheel-like appearance of rotavirus was visualized. Polyacrylamide gel electrophoresis was also applied on 15 of the positive samples; eight samples showed different polyacrylamide gel electrophoretic group A rotavirus long profile with different patterns. The results showed that the occurrence of rotavirus infection in cattle in Khartoum State is increasing.

Abstract Molar pregnancy is a rare condition in women, referring to the development of fetal membranes with or without an embryo. A 22-year-old mare referred to the veterinary faculty clinic due to the delayed parturition. In the ultrasound examination, the uterine membranes were seen suspended in fluids without any trace of the fetal organs. The mare underwent caesarean section, a mixture of brown fluid with fragments of fetal membranes was removed, but no fetus or any remnants of the fetus were found in the uterus. In the histopathological examinations, the fetal membranes showed severe degenerative changes. Also, the thickness of the endometrium had lost its normal structure and the layers of stratum spongiosum and stratum compactum could not be separated. The contents of the uterus were evacuated as much as possible and the mare regained her full health after the surgery. This is the first case of continued pregnancy without an embryo in a horse, which can be an unusual type of molar pregnancy.