Veterinary Research Forum

Abstract This study is the first to apply the behaviour, immunity, nutrition and environment (BIENE) protocol to assess dairy ewe welfare in intensive farms in Algeria. A total of 1,964 ewes from 50 farms in the Algiers region were evaluated between December 2022 and March 2023 using 13 animal-based indicators related to nutrition, housing, health, and behavior. Moderate tooth loss affected 54.67 ± 3.32% of ewes, and 20.38 ± 2.78% had a low body condition score (< 2). Housing problems included fleece moisture (11.28 ± 2.55%) and over 40.00% soiling of flanks and hindquarters. Health issues, such as mild anemia (23.55%), lameness (26.45%), nasal discharge (35.47 ± 4.43%), and labored breathing (8.79 ± 1.47%), were observed. In terms of behavior, most ewes appeared attentive and active, although 40.55% showed fear and 19.90% aggression. Flight time increased from 2.50 ± 0.07 sec to 3.80 ± 0.07 sec, and animals maintained greater distance from the corridor (2.79 ± 0.08 m) than from the observer (1.85 ± 0.06 m). Principal component analysis of qualitative behavior assessment identified three clusters, including well-being, discomfort, and vitality. Spearman correlation analysis showed that relaxed ewes had better body condition score (r = 0.29) and were less likely to be lame (r = – 0.32) or anemic (r = – 0.37). Vigorous ewes showed fewer signs of respiratory distress (r = – 0.32), while apathetic ones were more prone to poor conditioning (r = 0.31) and injuries (r = 0.30).The BIENE protocol proved effective in detecting major welfare issues in intensive Algerian sheep farming.

Abstract Porcine parvoviruses (PPVs) are globally recognized as significant contributors to reproductive failure in swine, primarily due to their association with fetal death. Infection in pregnant sows can lead to severe reproductive disorders including stillbirth, mummification, embryonic death and infertility. A recombinase polymerase amplification assay targeting the variable region of the outer capsid protein gene of the PPV-7 genome was developed and systematically optimized under a range of reaction conditions. The assay showed optimal amplification at a constant temperature of 35.00 ˚C for 25 min, using 0.72 µM of each forward and reverse primer and 14.00 mM magnesium acetate. It demonstrated high sensitivity, reliably detecting as few as 2,050 copies of viral nucleic acids in both the conventional and fluorescent dye-based formats. The assay also showed high specificity, exhibiting 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 positive for PPV-7, corresponding to a positivity rate of 13.77%. Operating at a low and constant temperature, the assay eliminates the need for advanced laboratory equipment, making it highly suitable for pen-side application in field settings. In conclusion, this novel assay demonstrated 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 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. The potential protective activity of gastrodin in rat testicular ischemia-reperfusion injury model and underlying mechanism were explored. Male rats were randomized into three groups, including sham control, testicular ischemia-reperfusion injury, and testicular ischemia-reperfusion injury along with gastrodin injection (n = 20). Testicular ischemia-reperfusion injury group received 2-hr period of left testicular torsion (720° and counterclockwise) and 4-hr or 3-month period of testicular detorsion. At the onset of testicular detorsion, gastrodin-treated rats were given 100 mg kg^-1 gastrodin by intra-peritoneal 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. Also, 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 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 three-dimensionel models based on computed tomography (CT) scan sections and stereology. In the study, seven adult female and seven male Koçeri sheep were used. The skulls used were scanned with a 64-detector Multi-Detector CT device with a section thickness of 0.625 mm, 80.00 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 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 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 3 weeks. The semen was divided into five equal portions and mixed with an extender having different concentrations of canthaxanthin: 0.00 (control), 5.00, 15.00, 25.00, 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 sperm kinematic factors 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, 25.00, 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 Bovine tuberculosis 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 the Assam State Zoo, Guwahati, 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 the tissue smears from granulomatous lesions confirmed the presence of acid-fast bacilli. Microscopic examination of tuberculosis 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 Basic Local Alignment Search Tool for nucleotide analysis following Sanger dideoxy 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 Menthol is an organic compound commonly 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. The present 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 25.00 mL organ baths containing Tyrode’s solution. The baths were maintained at 37.00 ˚C and continuously aerated with a gas mixture of 95.00% O₂ and 5.00% CO₂. The tissues were subjected to various contractile agents, including potassium chloride at concentrations of 30.00 and 80.00 mM, carbachol chloride at 1.00 and 4.00 μM and barium chloride at 30.00 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 (23.00 and 200 μg mL^-1) caused a rightward shift in the Ca2+ concentration-response curves, similar to verapamil. These findings suggested that menthol spasmolytic action might be mediated through the modulation and inhibition of calcium channels. In conclusion, menthol effectively attenuated bovine ileal smooth muscle contractions in vitro, indicating its potential as a natural therapeutic agent for controlling gastrointestinal hyperactivity in cattle.

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 Polycystic kidney disease (PKD) is a hereditary disorder characterized by progressive renal cyst formation, leading to compromised kidney function and an increased risk of chronic kidney disease (CKD). This prospective, 18-month longitudinal study evaluated diagnostic and prognostic markers in 12 PKD-affected cats, utilizing ultrasound imaging alongside biochemical analysis. The study documented a 14.00% increase in cyst volume, a 7.00% enlargement in kidney length, and significant elevations in plasma creatinine, phosphorus, and blood urea nitrogen levels, underscoring their correlation with disease progression. Hematological analysis revealed progressive anemia associated with advanced CKD stages, further outlining the systemic impact of PKD. These findings emphasize the progressive nature of CKD in cats with PKD, despite diligent care provided by pet owners and veterinarians. Also, the importance of routine sonographic evaluation along with hematological and biochemical assessments is highlighted in our study to enhance early detection and improve outcomes in feline PKD. Future longitudinal studies with expanded cohorts are essential to validate these findings and refine diagnostic criteria for PKD-associated CKD.

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 (DMEM) groups. Experimental groups: fibroblast, macrophage, and co-culture. 10µl supernatant was obtained from the separate cultures of fibroblasts, and macrophages and co-culture were injected. All injections were made through rete testis. 35 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 DMEM group had slightly better recovery conditions than the TD group, but the co-culture group showed similar conditions to the TD. Overall, it can 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.