Veterinary Research Forum

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 The aim of this study was to investigate the effects of N-butyl-2-cyanoacrylate (NBC), a synthetic tissue adhesive and graft material, on the healing of experimentally induced joint defects in rats. Twenty healthy male Wistar rats with an average body weight of 250 - 300g were used in this experiment. In the experimental group, NBC was given into the induced defects (2.00 mm) and then, they were closed routinely. In the control group, no intervention was made on the defects and they were closed routinely. All animals were observed during 30 days. In the radiological evaluation, the significant differences were recorded from days 15 to 30 between groups. In the histopathological evaluation, the distributions of collagen II and transforming growth factor-beta immuno-positive cells were more intense in experimental group than control group. As a result, the radiological and histopathological evaluations indicated that using NBC in joint defect increased tissue compatibility of newly formed cartilage cells and collagen fibers. However, how NBC provides chondrocyte production and harmony between the damaged and surrounding tissues remains a subject that must be studied.

Abstract Foreign bodies in the respiratory tract (trachea/bronchus) are very rare conditions; therefore, it is not on top of the list of differential diagnoses in patients with chronic or severe cough, but its occurrence should not be considered unexpected. A 2-year-old male Shih Tzu terrier mix dog (9.20 kg) with a history of dyspnea for 6 days was referred to the Veterinary Teaching Hospital of Shiraz University, Shiraz, Iran. In the clinical examination, respiratory distress and pain during palpation of the neck area were observed. Radiology indicated a triangular radiopaque object (bone) in the trachea of the animal. The size of the foreign body, the weakness of the grasping forceps of the device, and the edges of the bone being stuck in the trachea caused tracheoscopy failure after 30 min of trying. Surgical procedure (tracheotomy) was effective to remove the foreign body using Noyes alligator tissue forceps. Aspirated foreign body is not a common condition. In the case of early recognition and immediate treatment of the aspirated foreign body, the prognosis is usually good.

Abstract Melanocytoma and malignant melanoma are the most typical eye tumors in dogs and cats. Due to the presence of melanin, the general appearance is often highly pigmented in a nodular formation with well-defined borders. Electrochemotherapy (ECT) is a therapy combining reversible electroporation and anti-neoplastic drugs to enhance their cytotoxic effects through increasing cellular uptake by the electroporated tumor cells. In this article, the use of adjuvant ECT for the treatment of canine ocular melanoma is reported. The pre-surgical exams (blood count, renal and liver functions, echocardiogram, and electrocardiogram) were within the normal range, as were the chest radiography and abdominal ultrasound without signs of metastases. On the day of the surgery, an excisional biopsy of the tumor was performed without safety margins by keratectomy associated with conjunctivectomy and adjuvant ECT in the surgical site and peri-tumoral region. The animal was followed for approximately 7 months and remained without evidence of tumor recurrence. A complete resolution of corneal opacification was also observed. To our knowledge, this is the first report of the use of ECT in an ocular neoplasm. From a comparative oncology perspective, this report opens the way for future therapeutic approaches in superficial ocular cancers in veterinary and human medicine.

Abstract Uveitis is a major cause of vision loss. Methotrexate (MTX) has been widely used in uveitis due to its relatively safe profile. The purpose of this study was to evaluate the effects of two different dosages of MTX via intra-vitreal administration for treatment of endotoxin induced uveitis (EIU) in an experimental model. Thirty-five healthy rabbits were randomly divided into four groups and all animals were tolerated intra-vitreal injections. The first group received normal saline (NS), the second group received normal saline plus Salmonella typhimurium lipopolysaccharide endotoxin (LPS), (NS+LPS), the third group received 400 μg MTX plus LPS (LPS+MTX 400) and the fourth group received 800 μg MTX plus LPS (LPS+MTX 800). Intra-ocular inflammation was evaluated by clinical examination scoring during 7 post-injection days and histopathological examination at the end of study. Kruskal-Wallis and Mann-Whitney U tests were used to compare the histopathological and clinical scores. According to the clinical examinations, all groups demonstrated higher uveitis score than group 1 on first post-injection day. Also, groups 2 and 3 showed greater uveitis score than group 4. On the third, fifth and seventh post-injection days, clinical uveitis score in groups 2, 3 and 4 was significantly higher than group 1. The mean histopathological inflammation intensity scores in groups 2, 3 and 4 were significantly higher than group 1. Single intra-vitreal injection of 400 μg and 800 μg of MTX did not show significant anti-inflammatory effects on EIU in rabbits.

Abstract Feline lower urinary tract diseases are known to be life threatening conditions in cats, especially when they occur as obstructive diseases in males. Early diagnosis and treatment is necessary, otherwise it may lead to death. A 3-year-old male Persian cat was referred to the clinic with a history of anuria, lethargy, loss of appetite and exploratory cystotomy 6 months ago due to urethral obstruction following urolithiasis. Urinary bladder was enlarged and painful on palpation and urine accumulation was observed in ultrasonography. Biochemical and hematological analyses revealed hypocalcemia, hyperphosphatemia and hyperkalemia and increase in blood urea nitrogen, creatinine, white blood cell (WBC), red blood cell (RBC) and hematocrit. Urine analysis showed a turbid appearance, protein 1+, blood 3+, pH reduction, increased WBCs and RBCs and presence of bacteria, calcium oxalate crystals and epithelial cells. Urine culture reveled Staphylococcus saprophyticus. Postoperatively, microscopic examinations of the urinary bladder biopsy showed pathological lesions of bacterial cystitis. Based on these findings, bacterial cystitis and urethral obstruction due to post-operative urinary tract infections were diagnosed. For treatment, electrolyte imbalances were corrected firstly, cystotomy was performed and a catheter was conducted into the urethra; then, urethra was flushed and obstruction was resolved. Ampicillin was effective in reducing the bacterial count in urine. Despite the fact that cystotomy is a common procedure in veterinary medicine, clinicians should be aware of its complications such as post-operative urinary tract infections.

Abstract Recently, matrix metalloproteinases (MMP), in particular the gelatinases MMP-2 and MMP-9, generally considered as tumor markers for clinical applications. A longitudinal 2-years follow-up survey was performed on dogs with cutaneous tumor. Serum samples were obtained from 22 dogs with different cutaneous tumors and 22 health dogs at the time of surgery and one month, three months and one year after surgery. Gelatin zymography, hematological and biochemical assessment were performed for all serum samples. The serum alkaline phosphatase activity in dogs with malignant tumors was significantly higher than that in dogs with benign tumors and control cases. Latent forms of MMP-2 and MMP-9 were detected in all of the tumor cases. Gelatin zymography showed active form of MMP-9 in 12 cases (three benign and nine malignant tumors) and active form of MMP-2 in one fibrosarcoma case. Serum activity of active-MMP-9 and total MMP-9 was significantly higher in dogs with cutaneous tumors than those in controls. Tumor cases had higher serum activity of active-MMP-9 rather than controls. MMPs and alkaline phosphatase activities in serum were decreased significantly after surgery. Only one case with perianal gland adenoma showed recurrence of tumor four months after surgery in which active form of MMP-9 had identified one month before recurrence. According to the findings, it will be useful to measure ALP, MMP-2 and MMP-9 activities in the serum of dogs with cutaneous tumor for determination of tumor behavior before surgical treatment.

Abstract Tendon never restores the complete biological and mechanical properties after healing. Several techniques are available for tissue-engineered biological augmentation for tendon healing like stem cells. Recently, synovium has been investigated as a source of cells for tissue engineering. In the present study, we investigated potentials of fibroblast like synoviocytes (FLSs) in tendon healing. Sixteen rabbits were divided randomly into control and treatment groups. One rabbit was used as a donor of synovial membrane (synovium). The injury model was unilateral complete transection through the middle one third of deep digital flexor tendon (DDFT). Subsequently, the tendon stumps were sutured with 3/0 nylon. In treatment group, 0.1 mL phosphate-buffered saline (PBS) solution containing 1 × 10⁶ nucleated cells of FLSs was injected intratendinously at both tendon stumps just next to incision line. In control group, 0.1 mL PBS without FLSs was used for intratendinous injection. Model animals were euthanized at eight weeks, DDFTs were harvested and prepared for biomechanical study. Results of study showed that, there was no significant differences in biomechanical parameters values between FLSs treated and control groups. In conclusion, intratendinous injection of FLSs did not improve biomechanical properties during eight weeks in rabbit.

Abstract In the present study, we described cyto-histopathological features and immunophenotyping of the large B-cell lymphoma in an 8-year-old mixed breed dog with applying the World Health Organization (WHO) system of classification of canine lymphomas. In fine-needle aspiration (FNA), lymph nodes were involved by neoplastic cells of intermediate to large size with deep blue cytoplasm; consist of centroblasts, immunoblast and medium-sized cells. Histopathologically, the follicles and sinuses of lymph nodes were replaced by sheets of numerous immunoblasts (less than 90.0% of total cells) and centroblasts. Numerous mitotic figures were also observed. Immunohistochemical analysis presented that the neoplastic cells express B-cell phenotype CD20 and CD79a, but do not stain for T phenotype CD3. On the basis of cytology, histopathology and immunohistochemical findings, the present tumor was diagnosed as diffuse large B-cell lymphoma, high-grade centroblastic type (DLBCL-CB) according to WHO histological classification. Applying this classification system for diagnosis of canine lymphomas is very useful and has a high accuracy and consistency. However, further co-operative studies between clinicians and pathologists should be performed, in order to improve the effectiveness of this classification.

Abstract The aim of this study was to determine whether anesthesia consisting of sedation induced by intramuscular administration of xylazine-diazepam and lumbosacral analgesia induced by epidural administration of lidocaine and xylazine is satisfactory for castration and ovariohysterectomy in cats. Six adult (3 male and 3 female, 2.5 ± 0.5 years of age) cats (mean body weight ± SD, 2.2 ± 0.44 kg) were used in this study. Cats were sedated with xylazine (1-2 mg kg-1 IM) and diazepam (0.2 mg kg-1, IM) and 5 minutes later a 2% solution of lidocaine (0.5ml/4.5kg) and xylazine (1 mg kg-1) were administered into the lumbosacral epidural space. Open castration technique or ventral midline routine ovariohysterectomy were performed. Time to onset, duration and cranial spread of analgesia were recorded. Heart rate, respiratory rate and rectal temperature were recorded at time 0 (prior to epidural drugs administration) as a base line values and at 10, 20, 30, 45 and 60 minutes after the epidural administration. Onset time of analgesia was 4.0 ± 0.63 min (Mean ± SEM) and duration of analgesia was 89.5 ± 3.0 min (Mean ± SEM). However, surgical procedures were completed within 25-37 min. There were significant decrease in heart rate and rectal temperature values and significant increase in respiratory rate (P < 0.001). Intramuscular administration of xylazine-diazepam for sedation and epidural administration of lidocaine and xylazine for analgesia provided satisfactory analgesia for castration and ovariohysterectomy in cats. Utilizing epidural anesthetic technique with this combination is most useful for spaying surgery, especially when the surgical procedure can be completed in < 40 minutes.