Seasonal and Age-Associated Pathogen Distribution in Newborn Calves with Diarrhea Admitted to ICU.

Calf mortality constitutes a substantial loss for agriculture economy-based countries and is also a significant herd problem in developed countries. However, the occurrence and frequency of responsible gastro-intestinal (GI) pathogens in severe newborn diarrhea is still not well known. We aimed to determine the seasonal and age-associated pathogen distribution of severe diarrhea in newborn calves admitted to the intensive care unit (ICU) of Erciyes University animal hospital over a year. Fecal samples were collected during the ICU admissions, and specimens were subjected to a diarrheal pathogen screening panel that included bovine coronavirus (BCoV), Cryptosporidium spp., ETEC K99+, and bovine rotavirus, using RT-PCR and conventional PCR methods. Further isolation experiments were performed with permissive cell cultures and bacterial enrichment methods to identify the clinical importance of infectious pathogen shedding in the ICU. Among the hospitalized calves aged less than 45 days old, the majority of calves originated from small farms (85.9%). The pathogen that most frequently occurred was Cryptosporidium spp. (61.5%) followed by rotavirus (56.4%). The frequency of animal admission to ICU and GI pathogen identification was higher during the winter season (44.9%) when compared to other seasons. Most calves included in the study were 1-6 days old (44.9%). Lastly, co-infection with rotavirus and Cryptosporidium spp. occurred more frequently than other dual or multi-infection events. This study was the first to define severe diarrhea-causing GI pathogens from ICU admitted newborn calves in Turkey.

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.

Antibacterial, Antiviral, and Self-Cleaning Mats with Sensing Capabilities Based on Electrospun Nanofibers Decorated with ZnO Nanorods and Ag Nanoparticles for Protective Clothing Applications.

The COVID-19 pandemic has clearly shown the importance of developments in fabrication of advanced protective equipment. This study investigates the potential of using multifunctional electrospun poly(methyl methacrylate) (PMMA) nanofibers decorated with ZnO nanorods and Ag nanoparticles (PMMA/ZnO-Ag NFs) in protective mats. Herein, the PMMA/ZnO-Ag NFs with an average diameter of 450 nm were simply prepared on a nonwoven fabric by directly electrospinning from solutions containing PMMA, ZnO nanorods, and Ag nanoparticles. The novel material showed high performance with four functionalities (i) antibacterial agent for killing of Gram-negative and Gram-positive bacteria, (ii) antiviral agent for inhibition of corona and influenza viruses, (iii) photocatalyst for degradation of organic pollutants, enabling a self-cleaning protective mat, and (iv) reusable surface-enhanced Raman scattering substrate for quantitative analysis of trace pollutants on the nanofiber. This multi-functional material has high potential for use in protective clothing applications by providing passive and active protection pathways together with sensing capabilities.

Serological Investigation of Occupational Exposure to Zoonotic Crimean-Congo Hemorrhagic Fever Infection.

OBJECTIVE: Crimean-Congo hemorrhagic fever (CCHF) is an acute and highly fatal disease. In this study, our aim was to compare and evaluate the prevalence of CCHF virus (CCHFV) antibody among occupational high-risk groups by using the enzyme-linked immunosorbent assay and draw attention to the occupational groups that are at high risk for CCHF infection in an endemic region for this zoonotic infection in Erzurum, Turkey. MATERIALS AND METHODS: The antibody levels against CCHFV were surveyed among slaughterhouse workers, animal breeders, and veterinarians. The study population was composed of 72 participants having direct contact with animals and 19 blood donors who were not in direct contact with animals. RESULTS: The overall rate of CCHF immunoglobulin G positivity in risk groups was found to be 6.94% (5/72). CCHFV antibodies were found in 4 (12.5%) individuals of the animal breeder group. This ratio was considered significantly higher compared with the healthy control group. CCHFV antibodies were found in only one person (4.0%) who was an abattoir worker. In the veterinarian group, all people were found negative. CONCLUSION: In our study, the variables showing important associations with the prevalence of anti-CCHFV antibodies were livestock breeding, rural areas, and age. It was concluded that our region is endemic with regard to CCHF infection and persons who had direct contact with animals are at high risk. Thus, these participants must take necessary measures to protect themselves from CCHF and should be trained by health authorities.

GENETIC DIVERSITY of OVINE HERPESVIRUS 2 STRAINS OBTAINED FROM MALIGNANT CATARRHAL FEVER CASES in EASTERN TURKEY.

Malignant Catarrhal Fever (MCF) is a generalized, definitive lethal disease affecting the epithelial and lymphoid tissues of the respiratory and digestive tract, mainly cattle and some wild ruminants such as deer, buffalo or antelope. The sheep-related form of MCF is known to be present in Turkey and is caused by ovine herpesvirus 2 (OvHV-2). The aim of this study was to reveal the genetic diversity of OvHV-2 strains obtained from MCF cases in Eastern Turkey where the livestock industry has an important impact on economic activities. For this purpose, RTA (Replication and transcription activator), FGARAT (formylglycineamide ribotide amidotransferase) and some of glycoprotein genes (Ov7, Ov8 ex2, ORF27 and Ov9.5) were investigated in blood samples from 24 cattles, clinically diagnosed with MCF. Genomic data of chosen samples were furthermore used to characterize and undergo combined phylogenetic analysis to determine possible alleles and subvariants. The results showed that high level of OvHV-2 diversity existed in selected genes and strains carrying allelic variants might circulate both in two geographically distinct regions and in a region itself. Moreover, three different OvHV-2 types and various subtypes were identified based on multi locus approach. This study provides important data to epidemiological research and thereby helps to determine the source of the virus and understand the spread of the disease.

Detection and first molecular characterisation of three picornaviruses from diarrhoeic calves in Turkey.

The involvement of picornaviruses in calf diarrhoea was evaluated by the analysis of 127 faecal samples collected from diarrhoeic calves during 2014-2016. Virus detections were carried out by PCR using generic or specific primer pairs. One-third of the faecal samples (33.86%) were found to be positive for one or more of the studied viruses. Bovine kobuvirus was detected in 22.83%, bovine hungarovirus in 11.02%, while bovine enterovirus 1 in 5.51% of the samples. The sequences of the PCR products indicated the existence of novel variants in all the three virus species. When comparing the partial sequences, the nucleotide sequence identities between our newly detected viruses and those previously deposited to the GenBank ranged between 76 and 99%. Phylogenetic analyses revealed a novel lineage within the species Hunnivirus A. Our findings suggest that these viruses should be regarded as possible aetiological agents of calf diarrhoea. Based on the newly determined sequences, we designed and tested a new generic PCR primer set for the more reliable detection of bovine hungaroviruses. This is the first report on the molecular detection of the presence of bovine hungarovirus, bovine kobuvirus and bovine enterovirus 1 in the faecal samples of diarrhoeic calves in Turkey.

Expression of p53 protein, Jaagsiekte sheep retrovirus matrix protein, and surfactant protein in the lungs of sheep with pulmonary adenomatosis.

Ovine pulmonary adenocarcinoma (OPA) is a naturally occurring cancer in sheep that is caused by the Jaagsiekte sheep retrovirus (JSRV). Because the pathologic and epidemiologic features of OPA are similar to those of bronchoalveolar carcinoma in humans, OPA is considered a useful animal model for pulmonary carcinogenesis. In this study, 3,512 lungs from various breeds of sheep were collected and macroscopically examined. OPA was identified in 30 sheep, and samples of these animals were further examined by histologic, immunohistochemical (p53 protein, surfactant protein A [SP-A], proliferating cell nuclear antigen [PCNA], JSRV matrix protein [MA]), and PCR methods. Papillary or acinar adenocarcinomas were detected microscopically in the affected areas. Immunoreactivity for p53 PAb240 was detected in 13 sheep, whereas p53 DO-1 was not detected in any of the OPA animals. PCNA immunoreactivity was recorded in 27 animals. SP-A and JSRV MA protein was immunopositive in all 30. JSRV proviral DNA was detected by PCR analysis in all of the lung samples collected from OPA animals. In addition, the pulmonary SP-A levels were increased in tumor cells. The results of this study suggest that PCNA and p53 protein expression may be useful indicators in monitoring malignancy of pulmonary tumors.

The diagnostic value of two commercially available human cTnI assays in goat kids with myocarditis.

BACKGROUND: Cardiac troponin I (cTnI) is a peripheral blood marker for myocardial damage. Because of the unavailability of goat-specific cTnI assays human cTnI assays may be validated for detection of myocarditis in goat kids. OBJECTIVES: The purpose of the study was to evaluate 2 commercially available human cTnI assays in goat kids with myocardial damage, and to determine the cTnI expression in cardiac muscle. MATERIALS AND METHODS: Plasma cTnI concentrations were measured in healthy goat kids (n = 7) and goat kids with myocardial damage (n = 8) using the Beckman Coulter Access Accu TnI and the Biomérieux Vidas Ultra. The results were correlated with gross necropsy and histopathologic findings, and cTnI immunhistochemistry in cardiac tissue. RESULTS: Macro- and microscopic findings confirmed myocardial damage in the myocarditis group. Mean plasma cTnI concentration was significantly higher in the myocarditis group than in the healthy control group (104.82 vs 0.02 ng/mL). The overall mean plasma cTnI concentration measured by Biomérieux Vidas Ultra (61.75 ng/mL, 95% CI: 19.55-103.95) was comparable to the mean measured by Beckman Coulter Access Accu TnI (50.08 ng/mL, 95% CI: 24.11-76.06), and cTnI concentrations measured by these assays were highly correlated (r = .977) with a -6.2% bias. Both assays were precise and accurate. CONCLUSION: The human-specific Beckman Coulter Access Accu TnI and the Biomérieux Vidas Ultra can be used for diagnostic confirmation of myocardial damage in caprine medicine.

Immunization of knock-out α/ß interferon receptor mice against high lethal dose of Crimean-Congo hemorrhagic fever virus with a cell culture based vaccine.

Crimean-Congo hemorrhagic fever (CCHF) is an acute tick-borne zoonotic disease. The disease has been reported in many countries of Africa, Asia, the Middle East, and in Eurasia. During the past decade, new foci of CCHF have emerged in the Balkan Peninsula, southwest Russia, the Middle East, western China, India, Africa, and Turkey. CCHF virus produces severe hemorrhagic manifestations in humans with fatality rates up to 30%. Vaccine development efforts have been significantly hampered by a lack of animal models and therefore, no protective vaccine has been achieved. Lately, IFN α/ß receptor deficient (IFNAR-/-) mice have been established as a novel small animal model of CCHF virus infection. In the present study, we found that IFNAR-/- mice highly susceptible to CCHF virus Turkey-Kelkit06 strain. Immunization with the cell culture based vaccine elicited a significant level of protection against high dose challenge (1,000 PPFU) with a homologous CCHF virus in IFNAR-/- mice.

Detection of bovine torovirus in fecal specimens from calves with diarrhea in Turkey.

Bovine torovirus (BToV), a member of the family Coronaviridae, is an established gastrointestinal infectious agent in cattle. In this study, we performed a survey to detect BToV in Turkey between 2009 and 2011 using 235 fecal samples from neonatal calves with diarrhea that were analyzed by the nested reverse transcription (RT) PCR method using primers located in the consensus sequences of the BToV membrane (M) gene. The BToV M gene was detected in 4.7 % (11/235) of the samples using the nested RT-PCR method. The nucleotide sequences of partial M fragments from the BToV isolates, including the newly identified Turkish isolates, showed more than 96 % identity. The result indicates that BToV is one of the pathogens that contribute to neonatal calf diarrhea cases in Turkey.

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.