Detection and molecular characterization of Clostridium perfringens, Paeniclostridium sordellii and Clostridium septicum from lambs and goat kids with hemorrhagic abomasitis in Turkey.

BACKGROUND: The pathogenic Clostridia cause neurotoxic, histotoxic and enterotoxic infections in humans and animals. Several Clostridium species have been associated with abomasitis in ruminants. The present study aimed to investigate the frequency, and the presence of virulence genes, of Clostridium perfringens, Paeniclostridium sordellii and Clostridium septicum in lambs and goat kids with hemorrhagic abomasitis. RESULTS: A total of 38 abomasum samples, collected from lambs and goat kids of 1 week to 1 month of age in different farms located in eastern Turkey between 2021 and 2022, were evaluated by histopathology, culture and PCR. At necropsy, the abomasum of the animals was excessively filled with caseinized content and gas, and the abomasum mucosa was hemorrhagic in varying degrees. In histopathological evaluation, acute necrotizing hemorrhagic inflammation was noted in abomasum samples. The examination of swab samples by culture and PCR revealed that C. perfringens type A was the most frequently detected species (86.84%) either alone or in combination with other Clostridium species. P. sordellii, C. perfringens type F and C. septicum were also harboured in the samples, albeit at low rates. Beta2 toxin gene (cpb2) was found in three of C. perfringens type A positive samples. CONCLUSION: It was suggested that vaccination of pregnant animals with toxoid vaccines would be beneficial in terms of protecting newborn animals against Clostridial infections. This study investigated the presence of clostridial toxin genes in abomasal samples for the first time in Turkey.

First report of Serotype-1 Marek's disease virus (MDV-1) with oncogenic form in backyard turkeys in Turkey: a molecular analysis study.

BACKGROUND: Marek's disease (MD) is a lymphoproliferative disease caused by Gallid alphaherpesvirus 2 (GaHV-2, MDV-1), which primarily affects chickens. However, the virus is also able to induce tumors and polyneuritis in turkeys, albeit less frequently than in chickens. RESULTS: This is the first study in Turkey reporting the molecular characterization of a MDV-1 strain detected in a flock of backyard turkeys exhibiting visceral lymphoma. Here, MEQ, vIL-8, pp38 and 132-bp tandem repeat regions, which are frequently preferred in the pathotyping of MDV-1, were examined. It was determined that the MEQ gene of MDV-1/TR-21/turkey strain obtained in the present study encoded 339 amino acids (1020 nt) and had four proline-rich repeat regions (PPPP). Based on the nucleotide sequence of the MEQ gene of the MDV-1/TR-21/turkey strain, a phylogenetic tree was created using the MEGA-X software with the Maximum Likelihood Method (in 1000 replicates). Our strain was highly identical (> 99.8) to the Italian/Ck/625/16, Polish (Polen5) and some Turkish (Layer-GaHV-2-02-TR-2017, Tr/MDV-1/19) MDV-1 strains. Also, nt and aa sequences of the MEQ gene of our strain were 99.1 and 99.41% identical to another Turkish strain (MDV/Tur/2019) originated from chickens. Sequence analysis of pp38 and vIL-8 genes also supported the above finding. The identity ratios of nucleotide and amino acid sequences of vIL-8 and pp38 genes of MDV-1/TR-21/turkey strain were 99.64-100% and 99.79-100%, respectively, when compared with those of the Polish strain. According to 132-bp tandem repeat PCR results, the MDV-1/TR-21/turkey strain had five copies. CONCLUSIONS: These results suggested that the MDV-1/TR-21/turkey strain obtained from backyard turkeys can be either very virulent or very virulent plus pathotype, though experimental inoculation is required for precise pathotyping.

Detection and molecular characterization of a highly oncogenic Marek's disease virus from vaccinated hens in Turkey.

Marek's disease (MD) is a highly contagious neoplastic disease of chickens associated with economic losses, often due to visceral lymphomas. The etiological agent is MD virus serotype 1 (MDV-1), also called Gallid alphaherpesvirus 2 (GaHV-2). Despite intensive vaccination, MDV is constantly evolving and maintaining its presence in the world. The aim of this study was to genetically analyze a highly oncogenic MDV/Tur/2019 strain obtained from a poultry farm in Turkey's Elazig province in 2019. Genes associated with viral pathogenicity and oncogenicity Marek's EcoRI-Q-encoded protein (MEQ), phosphoprotein-38 (pp38), and viral interleukin 8 (vIL-8) were selected for this purpose. The vIL-8 nucleotide sequence showed high similarity (100% identity) to some European (EU-1, Polen 5) and Asian (03 India, GADVASU-M2) MDV strains. The pp38 nucleotide sequence showed high similarity (100% identity) to some American (CU-2, JM/102W, RB1B) and European (MD70/13, ATE2539) MDV strains. There were no disrupted four-proline molecules (PPPP) within the transactivation domain of the MEQ. However, according to phylogenetic results, the MDV/Tur/2019 strain was included in cluster 2a alongside European MDV strains (Polish, Hungarian, Italian) with very virulent and very virulent plus pathotypes. In conclusion, we believe that the MDV/Tur/2019 strain obtained from turkey herpesvirus (HVT)-vaccinated chickens has a very virulent or very virulent plus pathotype. Although this result provides some clues regarding the virulence of this strain, in vivo studies are needed to achieve exact pathotyping. Further, combination of HVT with the CVI988 strain should be used for vaccination to provide the best protection, as highly pathogenic MDV strains can break sterile immunity against the HVT vaccine. Keywords: GaHV-2; Marek's disease; oncogenes; Turkey.

Effects of Ovarian Transplant on Anti-Müllerian Hormone, Kisspeptin-1, and Kisspeptin-1 Receptor Levels in Rats.

OBJECTIVES: Information is scarce on levels of kisspeptin-1 and the kisspeptin-1 receptorin females after ovarian transplant. In this study, our aim was to explore serum estradiol, anti-Müllerian hormone, kisspeptin-1, and kisspeptin receptor levels, along with kisspeptin-1-positive cell density, in ovaries from rats after ovarian transplant. MATERIALS AND METHODS: For this study, 28 female Sprague Dawley rats were divided into 4 groups, with sham surgery performed on rats in group 1 (control group). Group 2 rats had ovaries transplanted under the peritoneum, and group 3 rats had their own ovaries transplanted subcutaneously. Group 4 rats were maintained in an estrous state. Serum anti-Müllerian hormone, kisspeptin-1, estradiol, and ovarian kisspeptin receptor levels were determined using commercial enzyme-linked immunosorbent assay kits. Kisspeptin-1-positive cell densities in the ovaries were determined immunohistochemically.The ovaries were also examined histopathologically. RESULTS: Our statistical analyses showed that levels of kisspeptin receptors in the ovaries were lowest in the subcutaneously transplanted group (group 3; 628.57 ± 35.69 pg/mL). The highest serum anti-Müllerian hormone levels were found in the estrous group (group 4; 16.91 ± 2.12 ng/mL). Differences between groups in terms of serum kisspeptin-1 and estradiol concentrations were not statistically significant. CONCLUSIONS: Our findings suggest that, in rats, results were better in the peritoneum transplant group than in the subcutaneous transplant group. We also found that serum anti-Müllerian hormone levels were lower in the transplant groups than in the estrous group, although levels were not completely decreased to zero. These results support the finding that ovarian activities continue after transplant.

Expression of the anti-Mullerian hormone, kisspeptin 1, and kisspeptin 1 receptor in polycystic ovary syndrome and controlled ovarian stimulation rat models.

Polycystic ovary syndrome represents a significant cause of female infertility. The aim of this study was to investigate the expression of anti-Mul-lerian hormone (AMH), kisspeptin 1 (KISS-1), and kisspeptin 1 receptor (KISS1r) in rat models of polycystic ovary syndrome (PCOS) and controlled ovarian stimulation (COS). For this purpose, 28 rats were assigned into four groups. Estrus and Diestrus groups consisted of rats in estrus and diestrus phases, respectively, while COS and PCOS groups consisted of rats with induced COS and PCOS, respectively. The serum AMH, KISS-1, and estradiol levels, and ovarian KISS1r levels were analyzed by enzyme-linked immunosorbent assay. Furthermore, histopathological analysis of the ovary tissue was done and ovarian KISS-1 expression was determined by immunohistochemical assay. The results revealed that ovarian KISS1r levels were higher in the Estrus (1271.43±51.98 pg/mL) and COS (1191.43±85.67 pg/mL) groups, compared to Diestrus and PCOS groups. The highest level of AMH was found in the Estrus group (16.91±2.12 ng/mL). The results indicate that AMH had no effect on the development of COS and PCOS, while KISS-1 was found to affect the development of COS in rats.

A new animal model for uterine torsion and uterine ischemia-reperfusion studies, but not fetal hypoxia studies.

The aim of the present study was to develop a new animal model for use in uterine torsion, uterine ischemia-reperfusion, and fetal hypoxia studies in rats. A total of 14 pregnant rats on their 18th-19th gestational days were used. The animals were randomly divided into two groups: those undergoing the shame operation (group 1), and those in which a 360 uterine torsion was performed using a novel technique, which was corrected 6 hours later (group 2). Subsequently, seven female and seven male rat pups aged 1 month were separated from the mothers in each group. The female rats were monitored until puberty via measuring the vaginal apertures. The 1-month old male rats and the female rats on reaching puberty were decapitated and histopathological tests were performed on the dissected organs, including the cerebral, visceral and genital organs. At the end of the study, no differences were observed between the groups with regard to abortions, offspring death rates and congenital abnormalities. It was observed that the time to reach puberty in female rats born from mothers with uterine torsion was longer, but the difference was statistically insignificant. No microscopic lesions were detected in the cerebral, visceral or genital organs of the offspring. Accordingly, it was concluded that offspring of mothers with the uterine torsion were not affected, at least in the short term. It was generally concluded that this animal model is appropriate for use in uterine torsion and ischemia-reperfusion studies, but is not appropriate for fetal hypoxia studies.

Effects of platelet rich plasma on fascial healing in rats with fecal peritonitis

PURPOSE: To evaluate the effects of platelet rich plasma (PRP) on the healing of fascia wherein peritonitis has been created. METHODS: Twenty eight Wistar Albino rats were divided into four groups. Only a primary fascial repair following laparotomy was performed on Group 1, a primary fascial repair performed and PRP treatment applied following laparotomy on Group 2, and a fecal peritonitis created following laparotomy and a primary fascial repair carried out on Group 3. A fecal peritonitis was created following laparotomy and primary fascial repair and PRP treatment on the fascia was carried out on Group 4. RESULTS: TNF-α was found to be significantly lower in the control group (Group 1). It was detected at the highest level in the group in which fecal peritonitis was created and PRP applied (Group 4). TGF-ß was determined as being significantly higher only in Group 4. Histopathologically, the differences between the groups in terms of cell infiltration and collagen deposition were not found to be significant. CONCLUSION: When platelet rich plasma was given histologically and biochemicaly as wound healing parameters cellular infiltration, collagen accumulation, and tissue hydroxyiproline levels were not increased but neovascularization, fibroblast activation and TNF Alfa levels were increased and PRP accelerated wound healing.

Effects of platelet rich plasma on fascial healing in rats with fecal peritonitis

ABSTRACT PURPOSE : To evaluate the effects of platelet rich plasma (PRP) on the healing of fascia wherein peritonitis has been created. METHODS: Twenty eight Wistar Albino rats were divided into four groups. Only a primary fascial repair following laparotomy was performed on Group 1, a primary fascial repair performed and PRP treatment applied following laparotomy on Group 2, and a fecal peritonitis created following laparotomy and a primary fascial repair carried out on Group 3. A fecal peritonitis was created following laparotomy and primary fascial repair and PRP treatment on the fascia was carried out on Group 4. RESULTS: TNF-α was found to be significantly lower in the control group (Group 1). It was detected at the highest level in the group in which fecal peritonitis was created and PRP applied (Group 4). TGF-β was determined as being significantly higher only in Group 4. Histopathologically, the differences between the groups in terms of cell infiltration and collagen deposition were not found to be significant. CONCLUSION: When platelet rich plasma was given histologically and biochemicaly as wound healing parameters cellular infiltration, collagen accumulation, and tissue hydroxyiproline levels were not increased but neovascularization, fibroblast activation and TNF Alfa levels were increased and PRP accelerated wound healing.

Clinical, pathological, and immunohistochemical findings in bald eagles (Haliaeetus leucocephalus) and golden eagles (Aquila chrysaetos) naturally infected with West Nile virus.

Fifteen bald eagles (Haliaeetus leucocephalus) and 3 golden eagles (Aquila chrysaetos) were diagnosed with West Nile disease based on 1) presence of lesions in brain, eyes, and heart, 2) viral antigen detection in brain, eyes, heart, kidney, and/or liver by immunohistochemical staining, 3) detection of viral RNA in tissue samples and/or cerebrospinal fluid (CSF) by polymerase chain reaction, and/or 4) detection of West Nile virus (WNV)-specific antibodies in CSF by serum neutralization assay. West Nile virus-associated gross lesions included cerebral pan-necrosis with hydrocephalus ex vacuo (7/15 bald eagles), fibrin exudation into the fundus in 1 golden eagle, retinal scarring in 1 bald eagle, and myocardial pallor and rounded heart apex in 4 bald eagles. Histologic lesions included lymphoplasmacytic encephalitis, most prominently in the cerebrum (17 eagles), lymphoplasmacytic pectenitis and choroiditis (15 and 8 eagles, respectively), and myocarditis (12 eagles). West Nile virus antigen was detected in the majority of the eagles in neurons of the brain (cerebrum and cerebellum), and less commonly present in neurons of the retina, tubular epithelial cells of the kidney, and cardiomyocytes. West Nile disease was diagnosed in 2 bald eagles based on the presence of cerebral pan-necrosis and WNV-specific antibodies in the CSF despite lacking viral antigen and RNA. In conclusion, WNV infection causes a fatal disease in bald and golden eagles. A variety of gross and histologic lesions are highly suggestive of WN disease in most eagles. A combination of detection of viral antigen and/or RNA or virus-specific antibodies proved useful in confirming the diagnosis.

Protective effects of protocatechuic acid on TCDD-induced oxidative and histopathological damage in the heart tissue of rats.

2,3,7,8-Tetracholorodibenzo-p-dioxin (TCDD) is a highly toxic environmental contaminant that causes severe toxic effects in animal and human. In this study, we investigated the toxic effects of TCDD and the preventive effects of protocatechuic acid (PCA), a widespread phenolic compound, in the heart tissue of rats. For this purpose, 3-4 months old 28 rats with 280-310 g body weights were equally divided into 4 groups (control, TCDD, PCA, TCDD + PCA group). A 2 µg/kg dose of 2,3,7,8-TCDD and 100 mg/kg dose of PCA were dissolved in corn oil and given orally to the rats for 45 days. The results indicated that TCDD induced oxidative stress by increasing the level of thiobarbituric acid reactive substance and by decreasing the levels of glutathione, catalase, glutathione peroxidase and superoxide dismutase in the heart tissue of rats. In contrast, PCA treatment prevents the toxic effects of TCDD on oxidative stress. In addition, histopathological alterations such as necrosis and hemorrhage occurred in TCDD group, and PCA treatment partially prevents these alterations in heart tissue. In this study, it was concluded that TCDD exposure led to toxic effects in heart tissue and PCA treatment could prevent the toxicity of TCDD.

The effects of Kisspeptin antibodies on delayed estrus in rats.

In recent years, numerous studies have been conducted on the effects of Kisspeptin/GPR54 system on sexual cycle, which proved that this system regulated gonadotropin release through GnRH. This study aims to determine the effects of hyperimmune serum containing antibodies produced against Kisspeptin on the sexual cycle and GnRH receptors in rat pituitaries. To this end, five Wistar female rats were passively immunised using a hyperimmune serum obtained from two Wistar female rats against Kisspeptin 10. Another five rats were selected as the control group. Anti-Kisspeptin antibodies of the hyperimmunised rats in the serum were identified by ELISA. The sexual cycles of the rats were followed by the measurements of vaginal irrigations and the estradiol concentrations in their blood samples. The number of follicles and corpora lutea in their ovaries was determined through histopathological tests, and the GnRH receptors in their pituitaries were identified by immunohistochemistry. Consequently, strong seropositivity was detected in all passively immunised rats and the hyperimmune serum. However, no difference was found between the groups with regard to the number of estrous cycles observed, the interval between estrous periods, estradiol concentrations, the number of follicles and corpora lutea and immunohistochemical results.

Pathological and immunohistochemical findings in broilers inoculated with a low virulent strain of infectious laryngotracheitis virus.

This study was carried out to describe clinical, gross and histopathological findings in the respiratory tract in chickens infected intranasally with A96 strain of infectious laryngotracheitis virus (ILTV). In addition, the presence of ILTV antigens in formalin-fixed and paraffin-embedded larynx and trachea tissues was investigated with the immunoperoxidase (IP) method in the infected chickens. At various days of viral infection, nares, larynx, trachea, lungs and air sacs tissue samples of the infected chickens were obtained and fixed with formalin and embedded in paraffin. The cross sections were stained with hematoxylineosin, and the larynx and trachea sections were also stained with the IP method. Mild rales and gasping were observed in only 4 of 35 chickens. The virus caused mild inflammatory changes in the respiratory tract. It was shown that clinical, gross and histopathological findings were not specific for differential diagnosis of the disease. However, ILTV antigens were detected by the IP method in formalin-fixed and paraffin-embedded larynx and trachea sections. These results revealed that the study use of the IP method might be useful for the diagnosis of ILTV infections with non-specific lesions.