Miokarditis vezan uz slinavku i šap u sisajuće teladi.

Foot-and-mouth disease (FMD) can lead to myocarditis in young animals, but the age distributions of calves with myocarditis have not been described, nor the biochemical profile in these calves. In an area endemic with foot-and-mouth disease, calves less than 6 months of age in infected farms were examined for clinical lesions and abnormalities in respiratory rate, heart rate and heart rhythm. In total, 53 calves were identified to be suspected of having foot-and-mouth disease infection. In 6 calves myocarditis was suspected based on tachypnea, tachycardia and gallop rhythm. In these 6 calves, cardiac troponin-I (cTnI) and aspartate aminotransferase (AST) were significantly higher (P<0.0001), but the levels of Creatinine Kinase MB (CK-MB) and Lactate dehydrogenase (LDH) were not. These 6 calves died within 2 days and histopathology confirmed myocarditis. All calves with myocarditis were younger than 2-months old, suggesting that myocarditis caused by FMD is mainly found in very young suckling calves.Slinavka i šap može dovesti do miokarditisa u mladih životinja. Dosada nije opisana dobna raspodjela miokarditisa ni biokemijski profil u teladi oboljele od slinavke i šapa. U jednom području gdje se slinavka i šap javlja endemijski, telad mlađa od šest mjeseci bila je na zaraženim farmama klinički pretražena posebice na poremećaje u frekvenciji bila, disanja i srčanog ritma. Ukupno su 53 teleta bila sumnjiva na slinavku i šap. Sumnja na miokarditis postavljena je u šest teladi i to na osnovi tahipneje, tahikardije i galopirajućeg ritma. U te su teladi razine srčanog troponina-I (cTnI) i aspartat-aminotransferaze (AST) bile značajno više (P<0,0001), dok razine kreatinin-kinaze MB (CK-MB) i laktat-dehidrogenaze (LDH) nisu. Tih šest teleta uginulo je unutar dva dana te je u njih miokarditis bio potvrđen patohistološki. Sva telad s miokarditisom bila je mlađa od dva mjeseca, što upućuje na zaključak da se miokarditis uzrokovan virusom slinavke i šapa pretežito javlja u sisajuće teladi najranije dobi

Detection of exogenous Jaagsiekte sheep retrovirus in Turkey

The aim of this study is to detect the exogenous Jaagsiekte sheep retrovirus (exJSRV) in suspected cases of ovine pulmonary adenocarcinoma (OPA) from the Eastern Anatolian region of Turkey. Pathological examination and PCR were carried out with the lung, lymph nodule, brain, heart and liver tissues of four sheep with suspected OPA. Histology of the lung sections indicated the well-circumscribed, multifocal and unencapsulated gray to white masses (arrows) on the parietal surface of medial and caudal lobes. exJSRV was detected in all tissue samples except for brain by nested polymerase chain reaction (nPCR). In addition to the nested-PCR results, the presence of exJSRV into the clinical samples was confirmed with sequencing of two PCR-positive products for OPAV. This report highlights the first presence of exJSRV in the sheep suspected with OPA in Turkey. Furthermore, the results provide supporting evidence for the metastasis of exJSRV in extra-thoracic tissues

First molecular characterization of hypodermin genes of Hypoderma bovis and serodiagnosis of bovine hypodermosis with recombinant hypodermin C antigen and a synthetic peptide containing its linear B-cell epitope.

Hypodermins A (HA), B (HB), and C (HC) of warble flies are modulatory antigens involved in host inflammation and immune responses during migration of the warble fly larvae through host connective tissues. In the current study, molecular characteristics of the genes encoding HA, HB, and HC were revealed from cDNA constructs of third-instar larvae of Hypoderma bovis. The open reading frame (ORF) of each hypodermin gene was amplified with modified gene-specific primers, and the resulting PCR products were cloned into pGEM-T Easy Vector to produce recombinant plasmids (rHA, rHB, and rHC). The ORF sequences of rHA, rHB, and rHC genes are 705 bp, 771 bp, and 783 bp long and encode proteins of 234, 256, and 263 amino acids with predicted sizes of 25.74 kDa, 27.79 kDa, and 28.51 kDa, respectively. The rHC gene was subcloned into the pET 100/D-TOPO Expression Vector, and the recombinant HC was purified using affinity chromatography. Western blotting indicated that rHC was recognized by the sera of cattle naturally infested with H. bovis. The rHC and a synthetic peptide (sHC) containing its linear B cell-specific epitope were evaluated as serological markers in indirect ELISA (iELISA) for the diagnosis of bovine hypodermosis. Both sHC and rHC iELISAs had sensitivity values equal to or higher than 90 % and specificity values of 100 %. A total of 200 serum samples from cattle in the Central Anatolia Region of Turkey were also analyzed by rHC and sHC-iELISAs to reveal the seroprevalence of bovine hypodermosis. The results of both iELISAs were consistent with one another and revealed a hypodermosis prevalence of 62 %. Our study provides the first data on molecular characterization of hypodermin genes of H. bovis and indicates the efficacy of recombinant antigen and peptide-based iELISA for serodiagnosis of bovine hypodermosis

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 similar to 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 similar to mu g/L (95% confidence interval [CI], 61.90348.06) and 0.013 similar to mu g/L (95% CI, 0.0100.017), respectively (t-value 19.27; P similar toBackgroundCardiac 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.ObjectiveThe 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.MethodsTen 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.ResultsAll lambs with myocarditis died within 1&nbsp;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&nbsp;&mu;g/L (95% confidence interval [CI], 61.90&ndash;348.06) and 0.013&nbsp;&mu;g/L (95% CI, 0.010&ndash;0.017), respectively (t-value 19.27; P&nbsp;&lt;&nbsp;.0001).ConclusionsA 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.</p

Sheep-Associated Malignant Catarrhal Fever: First report in a Calf in Northeastern Turkey

In this report, systemic vasculitis was described in a Brown Swiss calf with sheep-associated malignant catarrhal fever. The calf was referred to the university clinic due to respiratory and nervous symptoms. Nasal discharge, dyspnea, cough, conjunctival hyperemia, bilateral corneal opacity and ulceration, superficial lymph node enlargement, incoordination and muscle tremors were detected on clinical examination. The hematologic profile revealed lymphocytosis and neutrophilia. Grossly, hyperemia of viscera, lymphoid tissue enlargements and swelling in the brain were observed. Fibrinoid necrotic vasculitis and lymphoid cell infiltrations were main histopathologic changes in the brain, liver, spinal cord, heart, lymphoid tissues and upper respiratory tract. Characteristic histopathologic findings were confirmed by PCR, which demonstrated the presence of ovine herpesvirus-2 (OvHV-2) in the lymph nodes and liver samples of the calf

Confirmation of the antigenicity of the cell culture based vaccine by SDS-PAGE and western blotting.

<p>The unpurified vaccine antigens assessed before sucrose gradient ultracentrifugation and the purified vaccine antigens obtained after the purification and inactivation step were subjected to SDS-PAGE analysis shown in Fig. 2A and 2B, respectively. The purified vaccine samples were immunoblotted with 1:2000 dilution of the hyperimmune serum grown in rabbit against the CCHF virus and followed by goat anti-rabbit AP conjugated antibody to visualize the antigenic bands. (Fig. 2C). The cell culture based vaccine antigens were probed with anti-Gc (Fig. 2D) or anti-NP (Fig. 2E) monoclonal antibodies of the CCHF IbAr10200 virus.</p

Determination of virus titres of the vaccinated animals.

<p>Groups of 3 IFNAR^−/− mice were immunized three times at three weeks intervals with 5, 20, or 40 μg of the cell culture based vaccine. The control group of IFNAR^−/− mice (n = 3) was mock immunized with phosphate buffered solution (PBS). All animals were challenged with 1,000 PPFU (400 LD50) of CCHF virus Turkey-Kelkit06 strain two weeks after the last immunization. All animals were sacrificed at 2 days post-infection. Virus was isolated from the blood (A), liver (B) and spleen (C) as described in the Materials and Methods. Each point represents the mean values of the viral titre of three animals, and the standard deviations are shown as error bars.</p

Humoral immune responses to the cell culture based vaccine against CCHF in IFNAR^−/− mice.

<p>Groups of 6 IFNAR^−/− mice were immunized three times at three weeks intervals with 5, 20, or 40 μg of the cell culture based vaccine. The sera obtain from the mice at 14, 35, and 56 days were assayed for anti-CCHFV antibody end point titre (A) and neutralizing antibody titres determined by a 50% pseudo plaque reduction neutralization assay (PPRNT) (B). The error bars indicate the standard deviation. Differences in the ELISA titres and virus neutralization titres between groups were analyzed by 2-way ANOVA followed by Bonferroni’s post test, which is denoted by asterisks. *** (p<0.001).</p

Titres of the CCHF virus Turkey-Kelkit06 strain, weight loss curves, body temperatures changes and survival of IFNAR^−/−mice.

<p>Titres of the CCHF virus Turkey-Kelkit06 strain recovered in the blood (A), liver (B) and spleen (C) after intraperitoneal infection with 10¹, 10², 10³ or 10⁴ PPFUs of CCHF virus Turkey-Kelkit06 strain. All animals were sacrificed at 2 days post-infection (dpi). The virus was extracted from the blood and indicated organs, and the virus titres were determined by pseudo plaque assay (PPA). Each point represents the mean values of the viral titre of six animals, and the standard deviations are shown as error bars. The IFNAR^−/−mice (n = 6 each) were infected with 2.5, 5, 10¹, 10², 10³ or 10⁴ PPFUs of CCHF virus Turkey-Kelkit06 strain. The mice were monitored twice daily for mean weight change (D), body temperature (E), and survival (F). The median lethal dose was calculated by logistical regression. The standard deviations are shown as error bars.</p

Protection of IFNAR^−/− mice immunized with the cell culture based vaccine against CCHF virus Turkey-Kelkit06 strain challenge.

<p>Groups of 6 IFNAR^−/− mice were immunized three times at three weeks intervals with 5, 20, or 40 μg of the cell culture based vaccine. The control group of IFNAR^−/− mice (n = 6) was mock immunized with phosphate buffered solution (PBS). All animals were challenged with 1,000 PPFU (400 LD50) of CCHF virus Turkey- Kelkit06 strain two weeks after the last immunization. The mice were monitored twice daily for the cumulative mean symptom scores (A), the daily variations in weight as percentages compared to before the virus challenge (B), body temperature (C), and geometric mean time to death and survival (D). The animals were monitored for three weeks after the challenge. The standard deviations are shown as error bars.</p