Veterinary Research Forum

Abstract There is an increasing interest in exploring plant extracts as potential new treatment options for these ailments. Ziziphus vulgaris L. and Camellia sinensis (L.) Kuntze are medicinal plants widely used in different traditional medical practices, especially in Iranian Traditional Medicine, for addressing a range of health issues. Although their biological properties are well-established, their effectiveness against Giardia duodenalis cysts is still uncertain. This research aimed to assess the impact of Z. vulgaris and C. sinensis extracts on G. duodenalis cysts in vitro and to compare their effectiveness with metronidazole. Cysts were extracted from stool samples and concentrated using a 0.85 M sucrose solution. Extracts of Z. vulgaris and C. sinensis were prepared at concentrations of 25, 50, and 100 mg mL-1. The impacts of these extracts at different concentrations were evaluated at 10, 15, 30, 60, and 180 min, and the results were compared to control groups. The collected data were documented and statistically analyzed. The findings revealed that Z. vulgaris extract at a concentration of 100 mg mL-1and C. sinensis extract at a concentration of 50 mg mL-1 showed similar effectiveness to metronidazole in eliminating Giardia cysts. In addition, the cytotoxic effects of Z. vulgaris and C. sinensis extracts, in comparison to metronidazole, indicated a rise in fatality rates with prolonged exposure times and higher extract concentrations (p < 0.05). Therefore, it can be concluded that extracts of Z. vulgaris and C. sinensis are as effective as metronidazole for killing Giardia cysts in vitro.

Abstract This study evaluated the protective effects of intraperitoneal tannic acid (TA) against ischemia-reperfusion (I/R) injury in a rat model of testicular torsion. Eighteen adult male Wistar rats were randomized into three groups (n=6 each): sham (surgery without ischemia), I/R (3 hr ischemia + 3 hr reperfusion), and I/R+TA (TA 50 mg kg-1; 100 µL i.p. 30 min before reperfusion). Testicular tissues were sampled immediately after reperfusion for biochemical assays to measure malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione peroxidase (GPx) levels. Epididymides were analyzed 60 days later for sperm count, motility, viability, morphology, and DNA integrity. I/R significantly elevated MDA and sperm DNA damage while reducing SOD, GPx, sperm parameters, testicular weight and spermatogenesis. TA administration reversed these changes and restored the parameters to levels close to those of the sham group. Overall, intraperitoneal TA mitigated I/R-induced oxidative stress and preserved reproductive function, indicating its potential therapeutic value in testicular torsion.

Abstract Testicular ischemia-reperfusion (I/R) injury during testicular torsion is strongly influenced by oxidative stress caused by excessive accumulation of uncaptured reactive oxygen species (ROS). Kisspeptin-10, a biologically active fragment of the kisspeptin peptide family, has demonstrated significant antioxidant and anti-apoptotic properties. Recent studies indicate that kisspeptin-10 can mitigate oxidative stress by reducing reactive oxygen species levels and enhancing the activity of endogenous antioxidant enzymes such as superoxide dismutase (SOD) and catalase. This study examined the effects of kisspeptin-10 on I/R injury in testicular torsion/detorsion (T/D) of male rats. Twenty male rats were divided into four groups: the control group and three treatment groups (720° T/D, 720° T/D+ 0.50 µg kg-1 kisspeptin-10, 720° T/D+ 1.00 µg kg-1 kisspeptin-10). After inducing 720° clockwise testicular torsion for 2 hr, various factors such as sperm parameters, number, total motility, viability, DNA damage and hypoosmotic test were evaluated. The results showed that 720° T/D can increase sperm DNA damage. In addition, it also had negative effects on overall motility and other properties such as viability and plasma membrane functionality. The results also showed that administration of kisspeptin-10 to T/D rats can reduce DNA damage. These effects could also increase overall motility, viability and plasma membrane functionality compared to the T/D group. Based on our results, kisspeptin-10 provides significant protection against acute T/D injury to the testis when administered after spermatic cord torsion in rat.

Abstract Candida albicans is a commensal opportunistic yeast colonizing the skin and mucosal surfaces of humans and animals, which, under specific predisposing conditions, can proliferate excessively and lead to clinical manifestations of candidiasis. The widespread prophylactic and therapeutic use of antifungal agents has led to the emergence of drug-resistant strains, necessitating the exploration of novel therapeutic alternatives. Considering the clinical and conformational significance of equine skin health, the present study aimed to evaluate the therapeutic efficacy of ethanol extract of propolis in an experimental model of cutaneous candidiasis in horses. In 2022, two clinically healthy, six-year-old female horses (approximately 400 kg) were selected for the study. Immunosuppression was induced using dexamethasone and four intradermal inoculation sites were created on the shaved thoracic and flank regions of both lateral aspects using C. albicans (CPTC: 5027) suspension (5 × 10⁶ CFU mL-1). The experimental sites on the right thorax were treated with ethanol extract of propolis while those on the left thorax received topical nystatin. Lesions on the right flank were treated with glycerin as a vehicle control, and lesions on the left flank were left untreated. Cutaneous candidiasis was successfully induced within five days post-inoculation. Clinical resolution was observed following five days of treatment with ethanol extract of propolis, whereas, nystatin required eight days to achieve complete lesion resolution. The findings of this study suggested that topical ethanol extract of propolis demonstrated superior efficacy in accelerating the resolution of C. albicans-induced cutaneous lesions in horses compared to nystatin.

Abstract Corneal ulcers, particularly from alkaline chemical injuries, are a leading ocular complication in animals, accounting for about 20% of emergency referrals. These burns damage the conjunctiva and cornea, causing deep ulcers, potential perforation, delayed healing, infections, and blindness. Prompt treatment is crucial to accelerate healing and minimize complications like permanent opacity. This study compared the efficacy of blood serums (autologous, homologous, heterologous) against standard treatment (N-acetylcysteine and ciprofloxacin eye drops) and a control group (Normal saline) in treating induced corneal ulcers in two groups consisting of 10 healthy mixed-bred female dogs. Ulcers were created using 3M sodium hydroxide-soaked filter paper on both eyes. Evaluations at 1, 7, 14, and 21 days assessed quantitative parameters (corneal/stromal thickness, ulcer size) and qualitative symptoms (photophobia, blepharospasm, discharge, congestion, edema, neovascularization, opacity) via ultrasonography and clinical exams, respectively. Results showed significant time- and treatment-dependent improvements. Heterologous serum notably reduced thickness and ulcer size from day 7 compared to others. By days 14-21, all treatments (serums and drugs) lowered stromal thickness versus control. Qualitative parameters improved significantly with all treatments from day 7. These findings support the use of blood-derived serums, especially heterologous serum, to accelerate healing and improve clinical outcomes in canine corneal ulcers.

Abstract Bovine tuberculosis (bTB) is a chronic bacterial disease primarily caused by Mycobacterium bovis, a member of the Mycobacterium tuberculosis complex (MTBC), with significant zoonotic implications. This study aimed to detect MTBC in wildlife species, specifically Nilgai (Boselaphus tragocamelus) and Sambar deer (Rusa unicolor), using gross pathology, histopathology, acid-fast staining and molecular confirmation. Necropsied tissue samples were collected during post-mortem examination of a Nilgai and a Sambar deer from Assam State Zoo, India. Macroscopically, multiple granulomatous tubercles of varying sizes were observed in the lungs and liver, with creamy white caseous material marked upon sectioning. Ziehl-Neelsen staining of tissue smears from granulomatous lesions confirmed the presence of acid-fast bacilli. Microscopic examination of TB granulomas revealed a central necrotic mass surrounded by inflammatory cell infiltration, including Langerhans-type giant cells. Molecular confirmation of MTBC infection was achieved by amplifying hsp65 and IS1081 in tissue samples, further validated by nBLAST analysis following Sanger dideoxynucleotide sequencing. In conclusion, this study confirmed the presence of tuberculosis in these wildlife species through an integrated approach combining pathology, microbiology and molecular diagnostics, highlighting the need to understand pathogen entry into the herd and prevent potential spillover.

Abstract Abstract
The excessive production of reactive oxygen species during the freezing and thawing process triggers lipid peroxidation in the sperm membrane, resulting in oxidative harm and a decline in semen quality. The aim of this study was to examine the impact of different concentrations of canthaxanthin on the quality of goat spermatozoa throughout the process of cryopreservation. Semen was collected from each goat using an artificial vagina for a continuous duration of three weeks. The semen was divided into five equal portions and mixed with an extender having different concentrations of canthaxanthin: 0.00 µM (control), 5.00 µM, 15.00 µM, 25.00 µM, and 35.00 µM. The computer-assisted semen analyzer was used to determine the sperm motility and kinetic parameters. The findings indicated that the inclusion of canthaxanthin at a concentration of 5.00 µM resulted in a minimal enhancement in multiple factors associated with sperm motility, such as overall motility, progressive motility, fast progressive motility, curve-line velocity, distance curve line, amplitude of lateral head displacement, beat-cross frequency, wobble, linearity, and straightness. Nevertheless, these enhancements did not demonstrate statistical significance compared to the control, 15.00 µM, 25.00 µM, and 35.00 µM. The introduction of 5.00 µM of canthaxanthin resulted in a significant rise in straight-line velocity, average path velocity, and head activity compared to the control group. Hence, the addition of 5.00 µM of canthaxanthin could be employed to improve the cryopreservation of goat spermatozoa.

Abstract This study aimed to examine the effects of varying levels of L-carnitine and omega-3 and omega-9 fatty acids on performance, blood parameters, and testicular histomorphometry in fattening male Moghani lambs. Thirty-five five-month-old male Moghani lambs (27 ± 600 kg) were individually housed and randomly assigned to five dietary treatments. Diets included a control and four diets containing 3% calcium salts of omega-3 or omega-9 fatty acids with 100 or 200 mg/kg DMI of L-carnitine. Lambs were fed for 75 days to evaluate growth performance, nutrient digestibility, blood metabolites, carcass traits, and testicular histomorphometry.The results indicated significant effects on daily weight gain during the second month, average weight gain in the second month, and feed conversion ratio in the third month (P<0.05). However, glucose, total cholesterol, LDL cholesterol, triglycerides, blood urea nitrogen, and HDL cholesterol levels on day 30, as well as triglyceride levels on day 60, were not significantly influenced by dietary treatments. Blood urea nitrogen and HDL cholesterol levels, along with total protein, were significantly affected on day 60, with dietary treatments having a marked influence (P<0.05). Liver enzyme activities, including aspartate aminotransferase (AST) and alanine aminotransferase (ALT), were significantly affected during the first 30 days of the study (P<0.05), while alkaline phosphatase (ALP) activity remained unaffected over the 30 and 60-day periods. Testicular histomorphometry showed significant effects of dietary treatments on spermatogenesis index, seminiferous tubule diameter, and Leydig cell count (P<0.05).

Abstract Menthol is an organic compound derived from mint oils, known for its cooling and soothing properties, often used in various pharmaceutical, cosmetic, and therapeutic applications. It exerts beneficial effects on the digestive system by relaxing smooth muscles, reducing spasms, and improving gastrointestinal motility. This study aimed to evaluate the effects of menthol on bovine ileal smooth muscle contractions in vitro. Ileal tissue segments were collected from adult cattle at an abattoir and subsequently immersed in 25 mL organ baths containing Tyrode’s solution. The baths were maintained at 37 °C and continuously aerated with a gas mixture of 95% O₂ and 5% CO₂. The tissues were subjected to various contractile agents, including potassium chloride (KCl) at concentrations of 30 and 80 mM, carbachol (CCh) at 1 μM and 4 μM, and barium chloride (BaCl₂) at 30 mM. Menthol was cumulatively applied in incremental concentrations to assess its modulatory effects on contraction amplitude. Results demonstrated that menthol elicited a dose-dependent inhibition of smooth muscle contractions across most stimulatory conditions, with the extent of inhibition varying among different stimuli. The Ca2+ channel blocking activity was further confirmed when pre-treatment of isolated ileums with menthol (23 and 200 μg mL⁻¹) caused a rightward shift in the Ca2+ concentration-response curves (CRCs), similar to verapamil. These findings suggest menthol’s spasmolytic action may be mediated through the modulation and inhibition of calcium channels. Menthol effectively attenuates bovine ileal smooth muscle contractions in vitro, indicating its potential as a natural therapeutic agent for controlling gastrointestinal hyperactivity in cattle.

Abstract Porcine parvoviruses (PPVs) are globally recognized as significant contributors to porcine reproductive failure, primarily characterized by fetal death. It can cause infection in pregnant sows which can result in severe reproductive issues, including stillbirth, mummification, embryonic death, and infertility (SMEDI). An RPA assay targeting the variable region of the outer capsid protein gene of the PPV-7 genome was developed and systematically optimized under different reaction conditions. The assay exhibited optimal amplification performance at a constant temperature of 35 °C for 25 minutes, using 0.72 µM of each forward and reverse primer and 14 mM magnesium acetate (MgOAc). This assay demonstrated high sensitivity, detecting as few as 2050 copies of viral nucleic acids in both the conventional and fluorescent dye-based formats. The assay demonstrated high specificity, showing no cross-reactivity with other common porcine pathogens such as porcine sapelovirus, porcine circovirus, and classical swine fever virus.Of the 167 field samples tested, 23 were found to be positive for PPV-7, yielding a positivity rate of 13.7%. Operating at a low, constant temperature, the assay eliminates the requirement for advanced laboratory equipment, making it ideally suited for pen-side diagnostics in field conditions. In conclusion, this novel assay demonstrates strong potential for field-based detection of PPV-7 circulating within the swine population of Haryana, India, marking the first report of its kind from this region. Further validation using samples from clinically affected herds will strengthen its diagnostic applicability.

Abstract The present study was conducted to investigate the effect of L-carnitine (LC) supplementation in diets containing different levels of fish oil (FO) on performance, egg quality parameters and egg yolk fatty acids (FAs) profile in aged laying hens. In this study, 432 laying hens (Hy-line-W36, 65 weeks of age) were used allocated in a 3 × 3 factorial design with 6 replications and 8 birds per replication. The experimental treatments included diets containing 3 levels of FO (0, 1.5 and 3% of the diet) and 3 levels of LC (0, 300 and 600 mg/kg of the diet). The results showed that hens treated with 3% FO and 300 and 600 mg LC had the highest egg production rate, egg weight and egg mass, which was significantly higher than the treatments without FO and LC. With increasing usage level of FO to 3% of diet, egg yolk pH decreased significantly (p <0.05), however, yolk pH increased when diet supplemented with 600 mg LC. The percentage of polyunsaturated FAs (PUFA), the ratio of PUFA/saturated FAs (SFA) and the percentage of omega-3 FAs increased significantly with increasing FO usage level in the diet (p <0.05). Therefore, the use of LC in a diet containing FO not only increased egg production performance and egg internal quality (Haugh unit and yolk color) of aged laying hens, but also increased the production and deposition of omega-3 FAs in egg yolk.

Abstract The calculation of the cranial cavity volume is important for brain development, pathologies, and health status. In recent years, advancements in clinical applications have made volume calculation methods from radiological images more consistent. In this context, it was aimed to calculate the intracranial volume of the Koçeri sheep's skulls using 3D models based on computed tomography scan (CT scan) sections and stereology. In the study, 7 adult female and 7 male Koçeri sheep were used. The skulls used were scanned with a 64-detector Multi-Detector Computed Tomography device with a section thickness of 0.625 mm, 80 kV, 200 mA, and 639 mGy specifications. Using the MIMICS 20.1 software, three-dimensional models of the skull region were created and intracranial volumes were measured. The same reference points were measured using the Cavalieri principle. As a result of the study, measurements taken using CT scan showed that the cranial volume in females was 133.09 ± 7.81 cm³ and in males was 148.68 ± 16.25 cm³. The intracranial volume calculated using the Cavalieri principle was found to be 133.87 ± 8.43 cm³ in females and 149.77 ± 16.74 cm³ in males. The data were statistically analyzed in terms of two methods and sexual dimorphism. While no difference was found between the methods, a significant statistical difference was determined between the genders. In this context, our study is believed to make significant contributions to clinical sciences through accurate management of cranial disease diagnosis and processes, and to taxonomy through identification of interspecies differences.

Abstract Testicular ischemia-reperfusion is accompanied by elevated production of reactive oxygen species. It has been reported that reactive oxygen species are highly reactive to cellular carbohydrates, DNA, lipids and proteins, and result in testicular ischemia-reperfusion injury. Gastrodin is the principal active ingredient isolated from the medicinal plant Gastrodia elata Blume and has anti-oxidative stress effect. In this study, we explored the potential protective activity of gastrodin in rat testicular ischemia-reperfusion injury model and underlying mechanism. Male rats were randomized into 3 groups (sham control, testicular ischemia-reperfusion injury, testicular ischemia-reperfusion injury along with gastrodin injection) (n = 20). Testicular ischemia-reperfusion injury group received 2-hour period of left testicular torsion (720°, counterclockwise) and 4-hour or 3-month period of testicular detorsion. At the onset of testicular detorsion, gastrodin-treated rats were given 100.00 mg kg-1 gastrodin by intraperitoneal route. Following testicular detorsion, testicular tissues were collected for enzymatic activity analysis, oxidative stress evaluation and histopathological examination. The ipsilateral testicular xanthine oxidase activity (source of reactive oxygen species production) and malondialdehyde level (a precise biomarker of reactive oxygen species) were significantly increased in testicular ischemia-reperfusion injury group versus sham control group, while testicular spermatogenic function was decreased. Furthermore, gastrodin administration reduced xanthine oxidase activity and malondialdehyde level in ipsilateral testicular tissue, while improving testicular spermatogenic function. Consequently, it is suggested that gastrodin plays a protective role in testicular torsion/detorsion-induced ischemia/reperfusion injury through inhibiting xanthine oxidase activity to decrease reactive oxygen species formation.

Abstract Bone tissue engineering offers a potential alternative to conventional grafting by combining biocompatible scaffolds with biological stimulants. Polycaprolactone (PCL) is a biodegradable polyester with good mechanical strength and biocompatibility. Hydroxyapatite (HA) is a calcium phosphate mineral that is a major component of bone and has excellent bioactivity and biocompatibility. This research investigated the histopathological effects of polycaprolactone-hydroxyapatite (PCL-HA) nanocomposite scaffolds, along with their combination with platelet-rich fibrin (PRF), on the regeneration of bone in rabbit calvarial bone defects.
Four circular full-thickness bone defects of 5 mm in diameter were created on the calvarial bone of 15 male New Zealand white rabbits. Three defects were filled with PRF, PCL-HA, and PCL-HA/PRF, while one defect served as a control with no filler. Histopathological evaluations were conducted at 4, 8, and 12 weeks post-implantation. Data were evaluated using the Kruskal-Wallis and Mann-Whitney U tests.
Significant differences were observed between the treatment and control groups regarding bone regeneration throughout all three months studied. In the first month, no significant differences in bone regeneration were noted among the treatment groups. Still, in the second month, most new bone formation was observed in the PCL-HA/PRF group (p < 0.05). Both PCL-HA and PCL-HA/PRF groups significantly improved bone regeneration compared to control and PRF groups, with the PCL-HA/PRF group demonstrating the greatest bone formation, vascularization, and lowest inflammation by the 12th week. Thus, PCL-HA/PRF can be considered a suitable alternative to bone grafts and is increasingly utilized in orthopedic surgery and bone tissue engineering.

Abstract The present study evaluates the protective effects of royal jelly (RJ) on methotrexate (MTX)-induced testicular damage in rats, focusing on oxidative stress and autophagy. MTX, a folic acid analogue used in cancer and autoimmune treatments, impairs spermatogenesis via oxidative stress and apoptosis. Twenty-four male Wistar rats were divided into four groups: control (normal saline, 35 days), MTX (0.3 mg/kg, gavage, 3 times/week, 35 days), MTX+RJ (0.3 mg/kg MTX + 0.1 mg/kg RJ, gavage, 3 times/week, 35 days), and RJ (0.1 mg/kg, gavage, 3 times/week, 35 days). After 35 days, rats were euthanized, and testicular tissue was analyzed via histopathology, immunohistochemistry for LC3-I/II expression in germ cells, and qRT-PCR for mRNA expression of autophagy-related genes (Beclin-1, Atg7, LC3-I). Histopathological findings revealed that MTX caused severe interstitial edema, coagulative necrosis, and disrupted spermatogenesis, with reduced seminiferous tubule (ST) diameter, epithelial thickness, tubular differentiation index (TDI), and spermiogenesis index (SPI). Co-administration of RJ significantly improved ST morphology, diameter, epithelial thickness, TDI, and SPI (p<0.05). Immunohistochemistry showed a significant increase in LC3-I/II+ germ cells (spermatogonia, spermatocytes, spermatids) in the MTX group, which was markedly reduced in the MTX+RJ group (p<0.05). Similarly, qRT-PCR analysis demonstrated elevated mRNA levels of Beclin-1, Atg7, and LC3-I in the MTX group, which were significantly reduced in the MTX+RJ group (p<0.05). These findings suggest that RJ mitigates MTX-induced testicular damage by reducing oxidative stress and autophagy, thereby preserving spermatogenesis and testicular integrity.

Abstract Streptococcus agalactiae is a Gram-positive bacterial pathogen accountable for disease outbreaks in tilapia, resulting in substantial economic losses in aquaculture. In this study, a 10-round whole-cell Systemic Evolution of Ligands by Exponential Enrichment (SELEX) procedure was conducted to isolate and characterize single-stranded DNA (ssDNA) aptamers with specific binding affinity to S. agalactiae. The enriched ssDNA libraries were sequenced by high-throughput sequencing, yielding a total of 180,987 reads, of which 126,933 included unique aptamer sequences. The most frequent sequences were selected for further analysis. Among them, SA1 had the highest frequency, with 231 reads, whereas SA2 showed a lower number of 16 reads.
These findings underscore the validity of combining whole-cell SELEX, high-throughput sequencing, and aptamer-structure analysis to explore the interaction between aptamers and bacterial cell surfaces. The selected aptamers not only show promise for the detection of S. agalactiae but also serve as molecular probes for identifying surface-exposed proteins, studying host-pathogen interactions, and guiding targeted drug delivery in aquaculture-related applications.

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 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 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 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 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.

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 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.