Veterinary Research Forum

Abstract The present study evaluated the protective effects of royal jelly (RJ) on methotrexate (MTX)-induced testicular damage in rats, focusing on oxidative stress and autophagy. Methotrexate, a folic acid analogue used in cancer and autoimmune treatments, impairs spermatogenesis via oxidative stress and apoptosis. Twenty-four male Wistar rats were randomized into four groups: Control (normal saline, 35 days), MTX (0.30 mg kg-1, gavage, three times per week, 35 days), MTX + RJ (0.30 mg kg^-1 MTX + 0.10 mg kg^-1 RJ, gavage, three times per week, 35 days), and RJ (0.10 mg kg^-1, gavage, three times per 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 diameter, epithelial thickness, tubular differentiation index (TDI) and spermiogenesis index. Co-administration of RJ significantly improved seminiferous tubule morphology, diameter, epithelial thickness, TDI and spermiogenesis index. 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. 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. These findings suggested that RJ mitigated MTX-induced testicular damage by reducing oxidative stress and autophagy, thereby, preserving spermatogenesis and testicular integrity.

Abstract Crocetin (CRT) is one of the active chemical compounds of saffron and has many biological effects such as antioxidant property. The present study investigated the effects of CRT on crushed sciatic nerve function. Vitamin (Vit) C was used as an antioxidant drug. Thirty rats were divided into six groups including intact, sham, crush, CRT 7.50, CRT 30.00 and Vit C 100. Nine other rats with no surgery were scheduled in three groups to receive 7.50 and 30.00 mg kg^-1 CRT and 100 mg kg^-1 Vit C. In anesthetized rats, right sciatic nerve was crushed using a small hemostatic forceps. Sciatic functional index values on days five, 10, 15 and 20 after crush were accelerated, the severities of sciatic nerve degeneration and gastrocnemius muscle atrophy were ameliorated, and the increased malondialdehyde level and the decreased superoxide dismutase activity in the serum were restored by 20 consecutive days of oral administration of 30.00 mg kg^-1 CRT and 100 mg kg^-1 Vit C. No significant differences were observed between 30.00 mg kg^-1 and 100 mg kg^-1 Vit C. The groups that did not have surgery but received CRT (7.50 and 30.00 mg kg-1) and Vit C (100 mg kg-1) showed no behavioral, histopathological and biochemical alterations when compared to intact group. It was concluded that CRT and Vit C produced similar improving effects on crushed-injured sciatic nerve function. Inhibition of oxidative stress, enhancement of endogenous antioxidant activity might be involved in improving effects of CRT and Vit C.

Abstract This study, aimed to investigate Chlamydia abortus, C. pecorum, and C. psittaci in bovine, sheep, and goat abortions sent to the Konya Veterinary Control Institute diagnostic laboratory from various provinces in Central Anatolia and Mediterranean regions of Turkey between 2019 and 2022 via pathological and molecular methods. For this purpose, a total number of 150 abortions (from 58 cattle, 73 sheep, and 19 goats) were analysed via quantitative real-time polymerase chain reaction, histopathology and immunohistochemistry. The chlamydial DNA positivity rate was determined to be 24.00% via quantitative polymerase chain reaction analyses. According to the animal species, the positivity rate was 45.20% in sheep and 15.78% in goats. No cases positive for chlamydial DNA were found in the cattle abortions analyzed in the present study. When chlamydial agents were analysed on a species basis, C. abortus and C. pecorum were detected in 22.66and 1.33% of the cases, respectively. Among the animal species, 42.47% were C. abortus positive, whereas, 2.74% were C. pecorum positive in sheep. In the present study, the percentage of C. abortus-positive goats was 15.78%. and 22.66% of the Chlamydia spp. were positive according to immunohistochemical staining of aborted fetal lung, liver, heart, placenta and umbilical cord samples. This comparative study revealed that C. abortus was the most common chlamydial abortion agent in Türkiye. Chlamydial abortion agents cause abortion mostly in sheep in the region according to the animal species, and immunohistochemical staining contributes to the diagnosis of the agent at the Chlamydia spp. level.

Abstract The aim of this study was to investigate the presence of bovine herpesvirus type 1 and bovine parainfluenza virus type 3 (PI-3) in cattle lungs exhibiting pneumonia symptoms observed during macroscopic examination. Polymerase chain reaction, double immunohistochemistry, and double immunofluorescence methods were used for viral detection. For this purpose, during the summer-autumn period of 2023, a total of 84 lung samples from cattle aged 1 year and older, exhibiting macroscopic signs of pneumonia, were collected in Kayseri province, Türkiye. Pneumonia lesions were recorded according to the anatomical locations. In virus detection, PI-3 was identified at a rate of 8.33% using double immunohistochemistry/immunofluorescence staining methods, while this rate was found to be 6.72% in polymerase chain reaction. Bovine herpesvirus type 1 was not detected by any of the three methods. In the cattle lungs, 44 lesions (52.38%) were detected in the apical lobe, while seven lesions (8.33%) were detected in the middle lobe, eight lesions (9.52%) in the accessory lobe, and 25 lesions (29.76%) in the basal lobe. The PI-3 immunopositivity was observed in alveolar epithelial cells, as well as mononuclear cells in the interstitial and peri-vascular regions. This study is the first to examine bovine herpesvirus type 1 and PI-3 presence in bovine lungs using three distinct validation techniques (polymerase chain reaction, double immunohistochemistry, and double immunofluorescence).

Abstract A 4.5-year-old, nulliparous female New Zealand White Rabbit (Oryctolagus cuniculus) was presented for evaluation of a mammary mass located in right axillary mammary gland. The surgically excised mammary mass was submitted for histopathological and immunohistochemical evaluation. Histopathologically, neoplastic cells were arranged in sheets and nests of irregular sizes. The tissue was composed of amorphous eosinophilic material and necrotic cells. The neoplastic cells had round to oval nuclei with prominent nucleoli, scant basophilic cytoplasm and high mitotic activity. Cytokeratin and vimentin expressions of neoplastic cells were observed in immunohistochemical findings. Based on the microscopic characteristics and immunohisto-chemical staining, a diagnosis of a non-invasive comedocarcinoma was established. In this case, cytokeratin positivity confirmed the epithelial origin, whereas the vimentin positivity supported the presence of epithelial-mesenchymal transition and/or increased malignancy.

Abstract The efficacy of eucalyptol on testicular ischemia-reperfusion (I/R) injury remains uncertain. This study aimed to investigate the protective effects of eucalyptol on I/R injury induced by testicular torsion/detorsion (T/D) in rats. A total of 32 rats were divided into four groups, including control, T/D, E100 (T/D + 100 mg kg^-1 eucalyptol administered 30 min prior to torsion), and E200 (T/D + 200 mg kg^-1 eucalyptol administered 30 min prior to torsion). Testicular T/D was induced in the left testis via a 720° clockwise torsion for 3 hr, followed by 3 hr of detorsion. Testicular tissues were harvested for histopathological, immunohistochemical including cleaved caspase-3, B-cell lymphoma 2 (Bcl-2)-associated X protein (Bax), Bcl-2, and proliferating cell nuclear antigen (PCNA), and biochemical parameter as glutathione peroxidase (GPx), reduced glutathione (rGSH), glucose-6-phosphate dehydrogenase (G6PD), vitamin C, and malondialdehyde (MDA) analyses. In the T/D group, significant reductions in GPx, rGSH, G6PD, and vitamin C levels were observed, alongside increased MDA levels compared to the control group. Immuno-histochemically, the T/D group exhibited increased expression of cleaved caspase-3 and Bax, along with decreased expression of Bcl-2 and PCNA, compared to the control group. Histologically, Johnsen and Cosentino scores were irregular in the T/D group. Pre-treatment with eucalyptol resulted in reduced MDA, cleaved caspase-3, and Bax levels, while GPx, rGSH, vitamin C, G6PD, PCNA, and Bcl-2 levels increased. Additionally, improvements in Cosentino and Johnsen scores and histopathological damage were observed. These findings suggest that eucalyptol may exert a protective effect against I/R injury caused by testicular T/D, likely due to its anti-oxidant and anti-apoptotic properties.

Abstract The most significant infectious disease that affects cats is thought to be feline panleukopenia, also known as Cat distemper. Despite its epidemiological status, few literatures are available regarding the clinic-pathological aspect of the disease and about the molecular epidemiology of the circulating feline panleukopenia virus (FPV) in India. This study gives a comprehensive insight into the prevalence, pathology and diagnosis of FPV in cat population of Mizoram. Twenty-six cats that died of clinical disease suspected of FPV were subjected to a thorough pathological examination followed by molecular diagnosis. The FPV infection was confirmed in 12 out of the 26 cats by polymerase chain reaction assay targeting the VP2 gene of FPV. The phylogenetic analysis based on the full VP2 gene of FPV has demonstrated close genetic affinity of FPV strains circulating in Mizoram with the isolates from Thailand (MW589472), Italy (MZ508524) and China (OR727315). The analysis of the VP2-deduced amino acid sequence revealed two distinct mutations, S179T and I401V, exclusively identified in isolates from this particular study.

Abstract Doxorubicin (DOX), as a potent anti-cancer agent, exerts side effects in vital organs. Various chemical compounds with tissue protective properties are used to prevent the side effects of DOX. This study was planned to investigate the effects of histidine (HIS) and N-acetylcysteine (NAC) ​​on DOX-induced acute kidney injury. The possible mechanisms were followed by determining the histopathological changes of the kidney along with the biochemical alterations of the blood and kidney tissue. Forty-eight rats were divided into eight groups of six animals each to receive normal saline and DOX after alone and combined treatments with HIS and NAC. The DOX at a single dose of 15.00 mg kg^-1 was intraperitoneally injected on day one. The separate and combined intraperitoneally injections of HIS and NAC at a similar dose of 100 mg kg^-1 were began 30 min after DOX administration and continued for seven consecutive days. The DOX increased kidney weight and caused congestion, hemorrhages and degeneration in kidney tissue. It also increased serum urea and creatinine concentrations and kidney tissue levels of malondialdehyde, tumor necrosis factor-alpha and caspase-3, and decreased superoxide dismutase activity in this tissue. Separate and combined treatments with HIS and NAC improved all the above-mentioned effects of DOX. The restoring effects of the combined treatment were more prominent than the effect of amino acids alone. It was concluded that anti-oxidative, anti-inflammatory and anti-apoptotic mechanisms might be related to the tissue protective effects of HIS and NAC against DOX-induced acute renal injury.

Abstract This report presented a case of the concurrent diagnosis of early feline diffuse iris melanoma (FDIM) and metastatic cutaneous melanoma in a tricolored mixed stray cat. Histopathological examination of biopsy samples identified melanoma on the nasal planum, lips along with metastasis to the submandibular lymph nodes. The initial treatment included surgical resection of the primary nasal planum neoplasm and the metastatic lesions in the submandibular lymph nodes. The necropsy, performed 40 days after the surgery, revealed the metastatic nodules in multiple organs including the liver, lungs, thyroids, pericardium, pleura, peritoneum, kidneys, and numerous lymph nodes. Histopathological examination revealed a biphasic neoplastic tissue composed of a mixture of epithelioid and spindle cells in primary and metastatic tumors. Additionally, the anterior surface of the iris contained 4 - 5 rows of melanocytic interstitial infiltration indicative of early FDIM. Immunohistochemical analysis revealed that the neoplastic cells showed positive immunoreactivity for Melan-A. To the authors' knowledge, this case represented the first reported case of concurrent FDIM and cutaneous melanoma in a feline patient.

Abstract This study aimed to evaluate malondialdehyde (MDA) expressions using the immunohisto-chemical method in order to reveal the role of lipid peroxidation in the development and progression of hepatocellular carcinoma (HCC) induced by diethylnitrosamine (DEN) in male Wistar albino rats. Avidin-biotin-peroxidase method was used for immunohistochemical evaluation. Histopathological examinations revealed that DEN caused a mixed pattern (trabecular and acinar formations) of HCC in the majority of rats. The MDA positive stainings were significantly increased in rats in the HCC group compared to the healthy rats in the control group. In conclusion, this study data contain three important findings. The first one is that DEN triggers the formation of reactive oxygen species (ROS), excessively produced ROS cause oxidative stress, and as a result, oxidative stress strongly causes lipid peroxidation. Secondly, it is clear that there is an important relationship between oxidative stress-induced lipid peroxidation and HCC progression. At the same time, MDA is an useful biomarker in determining the prognosis of patients with HCC. The third and final finding is that intra-peritoneal DEN injection once a week for 20 weeks, but not in combination with other promoting chemical agents, appears to be very effective in inducing experimental HCC.

Abstract Erythropoietin (EPO) is a glycoprotein hormone predominantly produced in the kidneys, primarily stimulating erythroid cell proliferation in the bone marrow. The present study investigated the impact of EPO combined with swimming exercise on cardiac morphometry in adolescent male and female rats. The 4-week study involved 48 rats (24 males and 24 females), which were divided into four main groups of six males and six females each. The control group was administered intraperitoneal saline four times a week. The swimming exercise group also received intraperitoneal saline, followed by 30 min of swimming exercise, four times a week. The drug control group was given 50.00 IU kg^-1 epoetin alfa intraperitoneally, four times a week. Lastly, the Swimming + Drug group received 50.00 IU kg^-1 epoetin alfa intraperitoneally, four times a week, followed by 30 min of swimming exercise. The post-study measurements demonstrated that EPO administration did not result in notable alterations in crucial parameters, including the left ventricular mass index, left ventricular mass, and left ventricular posterior wall in the context of left ventricular hypertrophy in both genders. However, in female rats, EPO-only group and the combined EPO and exercise group showed significant thinning of the right ventricular wall and interventricular septum indicating potential cardiac dilatation. The results highlight the necessity of considering gender-specific responses when evaluating EPO's cardiovascular effects, particularly concerning the right ventricle, and suggest further investigation into the long-term consequences of these observed changes.

Abstract Schmallenberg virus (SBV) is a newly identified virus belonging to the Orthobunyavirus genus, of the Bunyaviridae family, and transmitted by haematophagous arthropods in particular mosquitoes and biting midges of the Culicoides genus. The SBV is known to cause reproductive disorders in ruminants mainly abortions, stillbirths and congenital malformations (hydranencephaly and arthrogryposis syndromes). The aim of this study was to investigate the presence of SBV in dairy cattle in Algeria. Between September 2023 and December 2023, blood serum samples from 300 dairy cows from 75 dairy farms in north eastern Algeria were tested for SBV antibodies using a commercial indirect enzyme-linked immunosorbent assay kit. Individual seroprevalence was 38.33% (115/300; 95.00% confidence interval: 32.83 - 43.83), while herd seroprevalence was 41.33% (31/75; 95.00% confidence interval: 30.18 - 52.47). In addition, the results of this study revealed that SBV seroprevalence at individual or herd level was high and not negligible confirming the presence of SBV in the regions studied in Algeria. In conclusion, more in-depth studies are recommended concerning the molecular proof, origin and pathogenesis of SBV in ruminants mainly those linked to reproductive disorders as well as the study of the various associated risk factors.

Abstract In this study, the effect of Gundelia tournefortii (GT) extract against diabetic gastropathy was investigated by pathological methods. The animal groups were designed as the control, diabetes, diabetes + GT50, diabetes + GT100, and diabetes + GT200 groups. No treatment was applied to the control group. The other groups received 45.00 mg kg^-1 streptozotocin intraperitoneally on the experimental day. The treatment groups were also given 50.00, 100, and 200 mg kg^-1 of GT extract daily by gavage for 21 days. Tissues were stained with Hematoxylin and Eosin for histopathological examination. Immunohistochemical staining was performed to reveal the presence of inflammation (tumor necrosis factor alpha), apoptosis (cysteine aspartate specific proteases-3), and oxidative stress (heat shock protein-27). Histopathological examination revealed no pathological lesion in the control group. In the diabetes group, mucosal tissue damage, and vascular and inflammatory changes were observed. In the treatment groups, GT decreased histopathological findings in parallel with the dose increase. Immunohistochemical examination revealed no immunopositivity in the control group, while severe immunopositivity was observed in the diabetes groups in terms of inflammation, apoptosis, and oxidative stress. In the treatment groups, there was a decrease in the severity of immunopositivity’s depending on the dose increase. As a result of this study, which has not been done before, GT was found to have a protective effect against gastropathy, being an important complication of diabetes, and this study is thus an important reference point for future research and promises new hope for the patients.

Abstract Infectious bronchitis, being caused by a coronavirus, is a significant disease affecting broiler and layer chickens, leading to substantial losses in the poultry industry due to the high mortality rates and decreased egg yield. Nearly 30 serotypes and 100 variants were described to date; developed vaccines are being for some severe cases, like the Massachusetts strain, to mitigate the effects. Determining the vaccinal strain's titer is crucial for creating an effective vaccine, and calculating the virus infectivity in the egg embryo is very important using dilutions ranging from 10^-3 to 10^-8, from each dilution 0.10 mL is used. The aim of this study was to determine the effects of the avian bronchitis virus injected into the allantoic cavity of ten days old embryonated eggs. Real-time polymerase chain reaction tests determined the viral load in the allantoic fluid. The embryos were removed to study gross injuries. The trachea, lung, and mesonephros were removed and submitted for histopathological studies, and nuclear factor-kappa B immunofluorescence analysis. The results revealed that the dilution of one-thousandth of the virus in the embryos caused the highest organ damage and viral replication. Varying degrees of hyperemia, edema, cellular infiltration, and degeneration were observed in the trachea, lung, and mesonephros depending on the virus dilution. This study provides valuable insights into the pathogenesis of the avian bronchitis virus, and has a potential impact on achieving an effective vaccine.

Abstract Porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus 2 (PCV2) are among one of the most prevalent and important infectious agents, which cause a severe economic impact on pig farming worldwide. Circulation of these viruses together in the same pig population may lead to the syndemic infection with altered pathogenicity and thereby pathology and clinical manifestation of the diseases. The present study evaluated syndemic infection between PRRSV and PCV2 in the pig population reared in the backyard farms in Mizoram state of India. The syndemicity between PRRSV and PCV2 was confirmed by clinico-pathological studies followed by simultaneous detection of both the viruses in tissue samples by polymerase chain reaction (PCR) and reverse transcription-PCR (RT-PCR), immunohisto-chemistry and SYBR green-based real-time PCR / RT- PCR. The syndemicity resulted to a more severe respiratory disease in affected pigs. Pathological studies on affected pigs revealed a combined picture of hemorrhagic lesions with lymphadenopathy. The SYBR green-based absolute quantification assay estimated the PCV2 load in more quantities than that of PRRSV in all the tissues. The highest PRRSV load was detected in the lungs, while the highest PCV2 load was detected in mesenteric and inguinal lymph nodes. The co-circulation of different pathogenic viruses and their persistent infections in a population is always a matter of great concern to pig producers across the world. The present findings highlighted the similar situation with syndemic infection of PRRSV with PCV2 for the first time in Mizoram, India, that caused severe respiratory disease in affected pigs.

Abstract Sudden death has been thoroughly well defined and investigated in human medicine and its definition is not as rigorously established in veterinary medicine. In this case a 7-year-old German Shepherd was brought to the Small Animals Clinic of the Faculty of Veterinary Medicine at the University of Belgrade in very poor state. Despite treatment, the dog died 15 min after being admitted to the clinic. Since poisoning was suspected, the owners requested an autopsy. The dog was taken to the Department of Forensic Veterinary Medicine, Faculty of Veterinary Medicine, University of Belgrade for necropsy. Pathomorphological examinations revealed rupture of the mesentery with mesenteric volvulus. In addition, a large number of adult forms of Dirofilaria immitis were found in the right atrium, ventricle and in truncus pulmonalis. Further examinations of the respiratory system revealed clusters with partially degraded adult parasites in the lobes. The dog died due to mesenteric volvulus which was a rare finding, with an even rarer finding that was lung nodules with adult D. immitis parasites.

Abstract An outbreak of contagious foot rot due to Fusobacterium necrophorum infection was reported in a herd of four-horned antelopes (Tetracerus quadricornis) in a wildlife sanctuary at Goa, India. The infected animals showed signs of anorexia, limping and foot lesions, like erosions and sloughing of hooves. Mortality was recorded in two antelopes in which gross lesions, like generalized congestion of skeletal muscles and internal organs, and multi-focal to coalescing abscesses in lung, pleura, and diaphragm were observed. Histopathological evaluation of interdigital skin showed zones of coagulative necrosis, infiltration of mononuclear inflammatory cells, congestion, hemorrhage, and thrombosis in the dermis. Lung showed extensive areas of consolidation with alveolar edema, hyperemic alveolar capillaries, hemorrhage, and infiltration of inflammatory cells. The bacterial culture from heart blood, lung, liver, and kidney tissues revealed isolation of F. necrophorum being also confirmed by polymerase chain reaction (PCR) using amplification of lktA gene of
F. necrophorum. In addition, the PCR also confirmed the presence of F. necrophorum DNA in the interdigital skin, lung, and liver tissues. Phylogenetic analysis showed that the
F. necrophorum isolate from the present study grouped closely with isolates from Australia, China, Pakistan, and Iraq, as well as an Indian isolate from goat. The affected live animals were treated with oral antibiotics, anti-inflammatory medicines, and vitamin supplements for seven days, and recovered. This study confirmed the first outbreak of contagious foot rot and systemic infection with F. necrophorum in a herd of four-horned antelopes in India.

Abstract Contagious caprine pleuropneumonia (CCPP) in goats is defined as a highly contagious and rapidly spreading mycoplasmal disease that is now among the leading causes of major economic losses on many continents (Asia, Africa and the Middle East). In this study, we aimed to evaluate immunohistochemically mast cells (MCs) profile and local interleukin (IL)-17 and IL-1β protein expressions in naturally infected CCPP according to the course of the inflammation (peracute-acute, subacute-chronic). The material of the study consisted of 40 naturally infected CCPP and 6 healthy control goat lung tissues. Appropriate samples were taken from the necropsied goats and subjected to histopathological and immunohistochemical examination. In the histopathological examination of the samples, it was determined that 29 samples had a peracute-acute course and 11 had a subacute-chronic course. In immuno-histochemical examination, MC profile and local IL-17 and IL-1β protein expressions were evaluated in the peracute-acute and subacute-chronic course. Immunohistochemically, significant increases in MC number, local IL-17 and IL-1β scores were detected in the peracute-acute course compared to the control group. There were significant decreases in the relevant scores in the subacute-chronic course compared to the peracute-acute course. Current findings indicated that MC, IL-17, and IL-1β expressions played important roles in the pathogenesis of infection in naturally infected CCPP, especially in the peracute-acute course. Additionally, MC profile was evaluated for the first time in naturally infected CCPP.

Abstract Hydropericardium syndrome (HPS) has caused significant financial losses to the Iranian poultry industry in the past few years. Thirty-two broiler chickens with gross lesions of HPS were inspected histologically and immunohistochemically. Sampling was performed in Sabzevar, Iran. The dead and sick birds from random farms were subjected to necropsy examinations. Only four broiler chickens had no hydropericardium and the other gross findings were similar for birds. Basophilic and eosinophilic intranuclear inclusions, hemorrhages and necrosis in different organs were the primary characteristic histologic lesions. Lymphoid depletion, goblet cell hyperplasia and necrotizing enteritis were some of the findings reported in previous research. Low macrophage infiltration rate and brain lesions were other discoveries in Hematoxylin and Eosin (H&E) examination. Feulgen reaction and Cluster of Differentiation 68 (CD68) immunohistochemical staining were used for a comprehensive investigation and these techniques revealed improved histopathologic details. Feulgen staining confirmed brain lesions and some other changes in different organs. Eventually, the CD68 method revealed low macrophage presence in most organs. This study suggested that HPS might cause brain damage and the susceptibility of the Arian breed to the adenovirus needs further investigation.

Abstract Prostate cancer is a very common cancer in men, affecting approximately 1.40 million men worldwide in 2020. To improve the quality of life and survival of both animals and humans, effective therapeutic approaches have been developed and evaluated using animal models. The rat model of prostate cancer induced by a multi-step protocol that consists of a sequential administration of flutamide, followed by testosterone propionate, then the administration of N-methyl-N-nitrosourea, and finally subcutaneous implantation of tubes filled with crystalline testosterone, is one of the most frequently used for prostate cancer research. However, the lack of standardization in procedures for prostate cancer induction, sample collection, and analysis represents a challenge for researchers. To address this issue, we aim to provide investigators with a detailed, step-by-step guide to implementing a rat model of prostate cancer, based on our extensive experience in this field. First, we briefly review the prostate cancer-induced protocols found in the literature, then we provide a detailed description of the prostate cancer rat model implemented by our team. After, we explore the rats’ prostate monitoring during the experiment protocol through imaging modalities, such as ultrasonography, computed tomography, and magnetic resonance imaging. We also describe animal welfare monitoring based on a table of humane endpoints, as well as data collection, such as biological variables and prostate samples. In sum, this article will ensure the quality of results and enable their comparison among different researchers using this rat model.

Abstract Squamous cell carcinoma (SCC) and more specifically tonsillar SCC are generally regarded as locally invasive with various rates of metastasis. A 13-year-old male Lhasa Apso was referred to the clinic with an unusual mass on his left tonsil and abnormal lumps under the tongue. The dog's food and water intake, breathing and heart rates, body temperature, capillary refill time, and blood tests were normal. A biopsy of abnormal tissue and immunohistochemical analysis were performed. Histopathological and immunohistochemical evaluations confirmed the basaloid SCC. Some treatments like surgery, radiotherapy, and chemotherapy are used to treat a specific tumor or area of the body. However, in this particular case, chemotherapy was not administered. Considering that dogs with tonsillar cancer generally have a poor to guarded prognosis and a short life expectancy, this case was euthanized after one year with the owner's consent due to the significant tumor enlargement.

Abstract A 16-year-old male mixed-breed dog presented with a mass with hemorrhage at the right conjunctiva. Five months after the initial visit, the right eye protruded and had a firm and irregular mass measuring approximately 1.00 cm in diameter with conjunctival hemorrhage. Microscopically, the mass was comprised polygonal or round tumor cells with distinct cell borders arranged in a nested and diffuse pattern. The tumor cells had round-to-oval fine hyperchromatic nuclei containing distinct multiple nucleoli and abundant eosinophilic or pale cytoplasm. Multiple giant cells were frequently observed. The mitotic index was 12.60/high power field. Extensive necrosis, hemorrhage and part of the cord-like and papillary epithelioid cells were observed in the intra-tumor tissue. Immunohistochemically, the tumor cells were positive for vimentin and α-smooth muscle actin and negative for cytokeratin, desmin and PNL2. On the other hand, the cord-like and papillary epithelioid cells were positive for vimentin, S100 and neuron-specific enolase. The tumor was diagnosed as an epithelioid leiomyosarcoma. This case considered to have occurred in the ocular region, although the ocular structure was destroyed.

Abstract Linguatula serrata can infect most ruminants and cause accidental infections in humans. It is a causative parasite of linguatulosis, a disease that not only produces economic losses in cattle but also represents a public health risk due to its zoonotic nature. This study aimed to explore the clinical and pathological findings of pulmonary linguatulosis in a rabbit. The most striking clinical findings in the deceased rabbits were wheezing and labored breathing. Grossly, the most prominent morphological changes in the lungs were well-circumscribed, flat or slightly raised, solitary grayish-white nodular lesions, and consolidated areas. The characteristically tongue-shaped developmental forms of parasites were observed on the cut surface of the lung. Histopathologically, the most noticeable morphological changes in the lung parenchyma were diffuse thickening of the inter-alveolar septum, fibrinoid necrotic vasculitis, medial smooth muscle cells hypertrophy of the arteries, alveolar emphysema, longitudinal and transverse sections of L. serrata nymphs and extra-medullary hematopoietic foci (megakaryocytes). The morphological appearance of the nymphs showed multiple transverse grooves, saw-like cuticles, peri-buccal hooks and acidophilic glands. In conclusion, these findings reveal the etiopathological diagnosis of linguatulosis and suggest that the lungs might be a target organ in addition to the liver and lymph nodes.

Abstract This study presents the first report of congenital tuberculosis in an 8-month-old male British Shorthair cat. The case was examined using histopathological and immunohistochemical methods. The cat was referred to a private veterinary clinic with general respiratory system problems and subsequent deterioration, leading to death. The cat owner granted permission for the cat necropsy and pathological examinations at Department of Pathology, Faculty of Veterinary Medicine, Selçuk University, Konya, Türkiye. During systemic necropsy, white round foci with diameters ranging from 3.00 to 5.00 mm were observed in the lung and spleen. Tissue samples were collected from the lung, spleen, liver, heart, kidney, mediastinal lymph nodes and brain for histopathological examinations. The tissues were subjected to routine histological tissue processing and sections were stained with Hematoxylin and Eosin and Ziehl-Neelsen. Additionally, Mycobacterium spp. antibodies were used for immunohistochemical staining. Microscopic examination revealed exudative tuberculosis lesions, areas of necrosis without a fibrous capsule and karyorrhectic cells only in the lung and spleen. Acid-resistant bacteria observed by ZN staining in the lesioned areas of the lung and spleen were identified as Mycobacterium spp. using immunohistochemical staining. No positive staining was observed in other organs using ZN and immunohistochemical methods. As a result, congenital tuberculosis was diagnosed in a cat for the first time, especially in relation to lesions in the spleen.

Abstract This study was aimed at the evaluation of cell proliferation, p53 level and apoptotic index by immunohistochemical methods in canine oral papillomatosis. The study material comprised of tumor tissue samples taken from six dogs being admitted to the Pathology Department of Faculty of Veterinary Medicine, Kafkas University, Kars, Türkiye. Choice of immunohistochemical staining was avidin-biotin peroxidase method. Cases of canine oral papillomatosis, determined to have been caused by canine papillomavirus-1, were found to have a rather high cell proliferation index. Furthermore, all cases were immunohisto-chemically demonstrated to carry a mutant p53 gene. Despite the mutation of p53 gene, the shift in the Bax/Bcl-2 ratio of dogs diagnosed with tumor was in favor of the pro-apoptotic Bax gene. The apoptotic mechanism was determined to occur through both the caspase-dependent and caspase-independent pathways. While the lesions occupied the entire oral cavity in some cases, histopathologically, malignant transformation was not detected in any of the six cases.