Astrovirus induced nonpurulent encephalomyelitis in sheep: First report from Türkiye by high-throughput sequencing.

BACKGROUND: This study presents the case of non-purulent encephalomyelitis associated with astrovirus infection in a sheep from Eastern Anatolia, Türkiye. METHODS: A necropsy was performed on a sheep showing nervous signs. Afterwards, brain tissue samples were taken and examined with histopathological, immunohistochemical and molecular techniques. RESULTS: Neuropathologic changes included neuronal degeneration, diffuse gliosis, multifocal perivascular cuffing, neuronophagy and neuronal necrosis in the cerebrum, the cerebellum and the cervical spinal cord. Aerobic and anaerobic bacterial culture, selective culture for Listeria monocytogenes, and PCR analysis for rabies virus, tick-borne encephalitis virus, Türkiye encephalitis virus, small ruminant lentiviruses and border disease virus were negative. However, the presence of astrovirus RNA in cerebral, cerebellar and spinal cord samples was demonstrated by a pan-astrovirus RT-PCR. Immunohistochemical examinations revealed astrovirus antigens within the neuronal cytoplasm. High-throughput sequencing techniques identified the causative agent as a member of the genotype species Mamastrovirus 13 but representing a distinct genetic lineage with similarity to ovine astrovirus 1 in the open-reading frames (ORF)1ab region and muskox astrovirus in the ORF2 region. CONCLUSION: This report provides evidence that astroviruses are potentially encephalitis-causing pathogens in ovine populations in Türkiye, featuring an astrovirus strain distinct from those previously identified in sheep.

Moelerella wisconsensis: first isolation from lungs and spleen of a horse infected with Streptococcus dysgalactia subsp. equisimilis.

Moellerella wisconsensis is a Gram-negative, facultative anaerobic bacillus of Entero-bacteriaceae family, and it is an uncommon pathogen in domestic animals. To date, five cases were reported including two dogs, two cattle, and a goat. Streptococcus equisimilis is the second common bacterial agent after the S. equi subsp. zooepidemicus in equine pneumonia cases. The present report describes the isolation of M. wisconses from lungs and spleen of a 10-year-old Arabian horse (May 08, 2022) at post-mortem examination being co-infected with S. equisimilis. Clinical and pathological findings included bilateral nasal discharge, conjunctivitis, sternal recumbency, severe diffuse necrosuppurative rhinitis, multi-focal fibrinopurulent pneumonia and purulent lymphadenitis. Polymerase chain reaction assays showed no viral nucleic acids of equid alphaherpesvirus (EHV) 1, EHV-4, equine arteritis virus and equine papilloma virus. The antibiogram test revealed that the isolate was sensitive to several antibiotics except colistin. Taken together, the present report documents the first isolation of M. wisconsensis from lungs and spleen of a horse; hence, experimental studies are needed to clarify the pathogenity and pathogenesis of M. wisconsensis.

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.

Evaluation of a collagen-bioaggregate composite scaffold in the repair of sheep pulp tissue.

PURPOSE: This study aimed to compare the effects of the collagen-BioAggregate mixture (CBA-M) and collagen-BioAggregate composite (CBA-C) sponge as a scaffolding material on the reparative dentin formation. MATERIALS AND METHODS: CBA-C sponge (10:1 w/w) was obtained and characterized by Scanning Electron Microscopy (SEM) and Mercury Porosimetry. Cytotoxicity of the CBA-C sponge was tested by using the L929 mouse fibroblast cell line. Dental pulp stem cells (DPSCs) were isolated from the pulp tissue of sheep teeth and characterized by flow cytometry for the presence of mesenchymal stem cell marker, CD44. The osteogenic differentiation capability of isolated DPSCs was studied by Alizarin Red staining. The cells were then used to study for the compatibility of CBA-C sponge with cell proliferation and calcium phosphate deposition. The effect of CBA-C sponge and CBA-M on the induction of dentin regeneration was studied in the perforated teeth of sheep for the eight-week period. All the analyses were performed with appropriate statistical hypothesis tests. RESULTS: CBA-C sponge was found to be biocompatible for DPSCs. The DPSCs seeded on the CBA-C sponge were able to differentiate into the osteoblastic lineage and deposit calcium phosphate crystals in vitro. Reparative dentin formation was observed after the second week in the CBA-C sponge applied group. At the end of eight weeks, a complete reparative dentin structure was formed in the CBA-C sponge applied group, whereas necrotic tissue residues were observed in groups treated with the CBA-M. CONCLUSION: CBA-C sponge represents a better microenvironment for reparative dentin formation probably due to maintaining DPSCs and allowing their osteogenic differentiation and thus calcium phosphate deposition.

Evaluation of cardiac troponin I in serum and myocardium of rabbits with experimentally induced polymicrobial sepsis.

Sepsis is a potentially life-threatening condition, and it is frequently complicated by myocardial damage. Data on myocardial damage in rabbit caecal ligation and puncture (CLP) models are limited, although numerous animal models have been used to study sepsis-associated myocardial damage. This study aimed to investigate the effect of CLP on cardiac muscle by measuring serum cardiac troponin I (cTnI) concentrations and by detecting both histopathological changes and cTnI immunoreactivity in cardiomyocytes in rabbits. After CLP was performed in rabbits, blood samples were taken from the jugular vein at 0, 4, 8, and 12 h for haematological and biochemical analyses. At the end of the experiment, all of the rabbits were euthanised to examine the histopathological changes and the cTnI immunoreactivity in cardiac muscle tissue. No changes in serum cTnI concentration were observed in the experimental group (EG) or control group (CG) at 0 and 4 h. In EG, the mean serum cTnI concentrations were 0.230 ± 0.209 and 1.177 ± 0.971 ng/ml at 8 and 12 h, respectively. In CG, the mean serum cTnI concentrations were 0.032 ± 0.014 and 0.031 ± 0.021 ng/ml at 8 and 12 h, respectively. Moreover, cytoplasmic cTnI immunoreactivity decreased in EG compared with that in CG (P<0.01). The results demonstrated that CLP induced a systemic inflammatory response and caused myocardial damage in rabbits.

Case report: systemic tuberculosis caused by Mycobacterium bovis in a cat.

BACKGROUND: The diagnosis of previous cases of feline tuberculosis in Turkey has been made based solely on pathological changes without isolation of the causative agent. This case report details the first case of feline tuberculosis in Turkey for which the causative agent (Mycobacterium bovis) was confirmed with microbiological isolation, morphological evaluation, molecular (PCR) characterization and antibiotic sensitivity. CASE PRESENTATION: Systemic tuberculosis was diagnosed via postmortem examination of a 5-year-old stray male cat. Mycobacterium bovis was isolated from the lungs, bronchial and gastrointestinal lymph nodes, kidney and liver. The isolate was defined as M. bovis using the Genotype MTBC assay (Hain Lifescience, Germany), which allows differentiation of species within the Mycobacterium tuberculosis complex with an easy-to-perform reverse hybridization assay. Pathological changes were characterized by multifocal to coalescing granulomatous inflammation in the lungs, liver, lymph nodes and kidneys. Further pathological changes included severe, diffuse, hepatocytic atrophy, periportal fibrosis with lymphohistiocytic infiltration, multifocal lymphohistiocytic interstitial nephritis, mild focal pulmonary anthracosis and mild renal and hepatic amyloidosis. Infection by immunosuppressive viral pathogens including feline herpes virus-1, feline immunodeficiency virus and feline parvovirus virus were ruled out by polymerase chain reaction assay (PCR). The isolated mycobacteria were susceptible to isoniazid, ethambutol, rifampicin or streptomycin. CONCLUSION: Disseminated M. bovis is a rare infection in cats. Involvement of submandibular lymph nodes suggested that primary transmission might have been the oral route in the present case.

The effectiveness of cyclooxygenase-2 inhibitors and evaluation of angiogenesis in the model of experimental colorectal cancer.

Colorectal cancer (CRC) is an important cause of cancer-related deaths worldwide. Early diagnosis and treatment of CRCs are of importance for improving the survival. In the present study, we studied the effects of nonsteroidal anti-inflammatory drugs (NSAIDs)-induced chemopreventive effects on tumor development incidence and angiogenesis in experimental CRC rats. 1,2-Dimethylhydrazine dihydrochloride (DMH) was used as cancer-inducing agent and two NSAIDs (celecoxib and diclofenac) were given orally as chemopreventive agents. Histopathological and immuno histochemical evaluations were performed in colorectal tissue samples, whereas angiogenesis parameters were studied in blood samples. Histopathological examination showed that adenocarcinoma (62.5%), dysplastic changes (31.25%) and inflammattory changes (6.25%) were detected in DMH group, whereas no pathological change was observed in control rats. In treatment groups, there was marked decrease in adenocarcinoma rate (30% and 10%, respectively). A significant increase was detected in MMP-2, MMP-9 levels and MMP-2/TIMP-2 ratio in DMH group as compared with controls and treatment groups. In immunohistochemical evaluations, there was an increase in intensity and extent of staining of MMP-2 and MMP-9 in DMH group as compared to controls and treatment groups. The decrease in celecoxib group was more prominent. Overall, it was concluded that NSAIDs, particularly cyclooxygenase-2 (COX-2) inhibitors, might have a protective effect on CRC development and slow down progression of tumor in a DMH-induced experimental cancer model. One of the possible mechanisms in the chemoprevention of colon cancer seems to be inhibition of angiogenesis by diclofenac and celecoxib.

Systemic nocardiosis in a dog caused by Nocardia cyriacigeorgica.

BACKGROUND: Systemic nocardiosis due to Nocardia cyriacigeorgica has not been reported in dogs. CASE PRESENTATION: Light and electron microscopy, microbiological culture and molecular identification (PCR) were used to diagnose systemic nocardiosis caused by Nocardia cyriacigeorgica in a 3-month-old husky dog. The postmortem changes included multifocal to coalescing, sharply circumscribed pyogranulomatous inflammation and abscess formation in lungs, liver, myocardium, spleen, kidneys, brain, and hilar lymph nodes. The organism was isolated and sequencing of its 16S rRNA allowed its identification and speciation. Examination of the bacterial culture by scanning electron-microscope showed filamentous branching with fragmentation into widely bacillary and cocoid forms of the bacteria. There was no history of immunosupressive drug administration and infection by the immunosuppresive viral pathogens, canine distemper and parvovirus were excluded via PCR. CONCLUSION: N. cyriacigeorgica should be considered potential cause of systemic pyogranulomatous lesions in dogs. It is the first reported case of systemic nocardiosis due to N. cyriacigeorgica in a dog.

Expression of Endoglin and Vascular Endothelial Growth Factor as Prognostic Markers in Experimental Colorectal Cancer.

BACKGROUND/AIM: Endoglin (CD105) is a receptor for the transforming growth factor-beta 1 (TGFß1) with crucial role in vascular development and angiogenesis. Additionally, vascular endothelial growth factor (VEGF) overexpression has been associated with advanced stage and poor survival for several cancer types. These molecules have been shown to be useful markers for identifying proliferating endothelium involved in tumor angiogenesis, especially in patients with cancer at risk of developing metastases. The aim of this study was to evaluate the relationship between VEGF and endoglin expression in an experimental model of colorectal cancer, as well as to investigate the effect of cyclooxygenase-2 (COX2) inhibitors on tumor development incidence. MATERIALS AND METHODS: Colon cancer was induced with 1,2-dimethylhydrazine dihydrochloride (DMH). Celecoxib and diclofenac treatment was started simultaneously with DMH induction. Endoglin protein expression was performed using western blot analysis. VEGF plasma concentrations were measured by enzyme-linked immunosorbent assay. RESULTS: In histopathological evaluations, no pathological change was observed in control rats, while adenocarcinoma (62.5%), dysplasia (31.25%) and inflammation (6.25%) were detected in the group given DMH. In treatment groups, a marked decrease was observed in adenocarcinoma rate. Expression of endoglin protein was significantly elevated in the DMH group compared to controls (p<0.001). No statistically significant difference was noted between treatment groups and DMH group regarding endoglin expression but a decrease was detected in the celecoxib-treated groups. CONCLUSION: It was confirmed by histopathology and western blotting that COX2 inhibitors, particularly celecoxib, decrease the rate of disease and slow-down progression of existing CRC. These data show that endoglin expression may have an important role in tumor angiogenesis and predict of tumor invasion.

First Report of Echinococcus equinus in a Donkey in Turkey.

A 2-year-old female donkey (Equus asinus) was euthanized in the Pathology Department of Firat University, Elazig, Turkey. Necropsy disclosed the presence of 7 hydatid cysts distributed throughout the lung parenchyma. One of those cysts represented the parasite material of the present study and was molecularly identified through sequencing of a fragment of cytochrome c oxidase subunit 1 (CO1) and nicotinamide adenine dinucleotide dehydrogenase subunit 1 (NADH1) gene, as Echinococcus equinus. The generated CO1 sequence supports the presence of the dominant haplotype as has been described in Europe and Africa. The NADH1 sequence was found similar to sequences reported in equids in Egypt and the United Kingdom. The molecular identification of E. equinus in a donkey is being reported for the first time in Turkey.

Identification by culture, PCR, and immunohistochemistry of mycoplasmas and their molecular typing in sheep and lamb lungs with pneumonia in Eastern Turkey.

This study used cultures, polymerase chain reaction (PCR), and immunoperoxidase to examine samples from 216 lungs from sheep and lambs with macroscopic pneumonia lesions for the presence of Mycoplasma species. DNA was extracted from lung tissue samples and broth cultures with the help of a DNA extraction kit and replicated using genus-specific and species-specific primers for mycoplasma. The lung samples were examined by the immunoperoxidase method using hyperimmune Mycoplasma ovipneumoniae serum. The randomly amplified polymorphic DNA (RAPD) test was used for the molecular typing of M. ovipneumoniae isolates. Mycoplasma was isolated in the cultures of 80 (37.03 %) of a total of 216 lung samples. Genus-specific mycoplasma DNA was identified by PCR in 96 (44.44 %) samples in broth cultures and 36 (16.66 %) directly in the lung tissue. Of these 96 cases in which genus-specific identification was made, 57 (59.37 %) were positive for reaction with species-specific primers for M. ovipneumoniae and 31 (32.29 %) for Mycoplasma arginini. The DNA of neither of the latter two species could be identified in the remaining eight samples (8.33 %) where mycoplasma had been identified. As for the immunoperoxidase method, it identified M. ovipneumoniae in 61 of 216 lung samples (28 %). Positive staining was concentrated in the bronchial epithelium cell cytoplasm and cell surface. RAPD analysis resulted in 15 different profiles. Our results suggest that PCR methods could be successfully used in the diagnosis of mycoplasma infections as an alternative to culture method and identifying this agent at the species level.

Increased plasma cardiac troponin I concentration in lambs with myocarditis.

BACKGROUND: Cardiac troponin I (cTnI) is a blood biomarker of myocardial injury. A human cTnI assay may be useful for measuring cTnI concentrations in lambs with naturally occurring myocarditis. OBJECTIVE: The aims of this study were to evaluate the utility of a commercially available human chemiluminescent microparticle cTnI immunoassay for measuring plasma cTnI concentrations in lambs with naturally occurring myocarditis from infection with foot and mouth disease virus (FMDV), and to determine cTnI expression in cardiac muscle of affected lambs. METHODS: Ten lambs with myocarditis and 10 clinically healthy lambs (control group) were included. Clinical signs, gross and histologic necropsy findings, and immunoreactivity for cTnI in cardiac tissue were evaluated. Plasma cTnI concentration was determined using the commercial human immunoassay system. RESULTS: All lambs with myocarditis died within 1 day of clinical signs. Infection with FMDV was confirmed by PCR analysis. Gross cardiac lesions were evident and histologic examination revealed myocarditis. Immunoreactivity for cTnI was absent in cardiac myocytes that were degenerative or necrotic, but was strong in cardiac myocytes from unaffected areas of the myocardium and in all cardiac myocytes of healthy lambs. The geometric mean plasma concentrations of cTnI for lambs in the myocarditis and control groups were 146.78 µg/L (95% confidence interval [CI], 61.90-348.06) and 0.013 µg/L (95% CI, 0.010-0.017), respectively (t-value 19.27; P < .0001). CONCLUSIONS: A commercial human cTnI assay may be used to detect plasma cTnI concentrations in sheep, and cTnI may be used as a blood-based biomarker of myocarditis in this species.