Group A rotaviruses (RVA) are regarded as major enteric pathogens of large ruminants, including cattle. Rotavirus vaccines administered to pregnant cows are commonly used to provide passive immunity that protects newborn calves from the clinical disease. In this study we report the detection of RVA from calves with severe diarrhea in a herd regularly vaccinated to prevent enteric infections including RVA. Diarrheic disease was observed in newborn calves aged 4-15days, with high morbidity and mortality rates, but no diarrhea was seen in adult animals. Rotavirus antigen was detected by enzyme-immunoassay in the intestinal content or the fecal samples of all examined animals. Besides RVA, bovine coronavirus and bovine enteric calicivirus were detected in some samples. Selected RVA strains were characterized by whole genome sequencing. Two strains, RVA/Cow-wt/TUR/Amasya-1/2015/G8P[5] and RVA/Cow-wt/TUR/Amasya-2/2015/G8P[5] were genotyped as G8-P[5]-I2-R2-C2-M2-A3-N2-T6-E2-H3 and showed >99% nucleotide sequence identity among themselves. This genomic constellation is fairly common among bovine RVA strains; however, phylogenetic analysis of the G8 VP7 gene showed close genetic relationship to some European human RVA strains (up to 98.4% nt identity). Our findings is the first indication regarding the circulation of G8 RVA strains in Turkey. Given that the administered RVA vaccines contained type G6 and G10 VP7 antigens some concerns raised with regard to the level of heterotypic protection elicited by the vaccine strains against circulating bovine G8 RVA strains. Enhancement of surveillance of circulating RVA strains in calves across Turkey is needed to support ongoing vaccination programs.
A calicivirus was detected in neonatal calves with enteritis in Kırklareli, Thrace, Turkey. In the full-length genome, Kırklareli virus was related (48% nucleotide identity) to bovine enteric caliciviruses (Nebovirus genus). The virus was also detected in a herd in Ankara, Central Anatolia, but not in other Turkish prefectures.C aliciviruses (family Caliciviridae) are important pathogens of humans and animals. Caliciviruses are nonenveloped, small round viruses with a single-strand positive-sense RNA genome of 7 to 8.5 kb in size, polyadenylated at the 3= end (1). The family includes the genera Vesivirus, Norovirus, Sapovirus, Lagovirus, and Nebovirus. In recent years, novel, yet unclassified caliciviruses have been identified in mammals, birds, and fishes (2).Caliciviruses of at least three distinct genera (Nebovirus, Norovirus, and Vesivirus) have been detected in cattle. However, on the basis of either experimental infections or observational studies, only neboviruses and noroviruses have been associated with enteric replication and with enteric signs (3, 4).In early 2012, an outbreak of enteritis occurred in Kırklareli, Thrace, Turkey. The herd consisted of 250 cows and 200 calves, with enteric disease affecting about 60% of the calves, resulting in 30% mortality. A total of 17 fecal samples, collected from the affected calves, were tested and found to be positive for either species A rotavirus or coronavirus. All animals also tested positive for Cryptosporidium spp. Interestingly, upon electron microscopy observation, small round viruses (SRVs) could be observed in 3 stool samples. Using reverse transcription (RT)-PCR with consensus primers (5) and the sequence analysis of a short (330-nucleotide [nt]) fragment of the RNA-dependent RNA polymerase (RdRp), the SRVs were characterized as caliciviruses, with limited homology (Ͻ65% nt identity), to the prototype strain Nebraska (Nebovirus genus) (6). By combining RT-PCRs with a primer walking strategy, 5= and 3= RACE (rapid amplification of cDNA ends) protocols and next-generation sequencing, the complete genome of the calicivirus, named Kırklareli virus, was determined (GenBank accession no. KT119483). The genome was 7,484 nt long, excluding the poly(A) tail, with a ribonucleoside composition of 20.4% A, 30.9% C, 27.5% G, and 21.2% U. The 5= untranslated region (UTR) was 67 nt long, while the 3= UTR was 80 nt long. Two open reading frames (ORFs), of 6,581 nt (ORF1) and 657 nt (ORF2) in length, were mapped. ORF1 encoded a large polyprotein of 2,226 amino acids (aa) in length, containing the replicative proteins and, at the 3= end, the 1,629 nt-long (542 aa) capsid region, with a potential cleavage site, EGD, between the nonstructural proteins and the capsid protein. ORF2 encoded a 218-aa long protein (Fig. 1). Conserved amino acid motifs characteristic of caliciviruses, including the 2C-like helicase-nucleoside triphosphatase DNA-binding (GXXGXGKS/T) and the ATP hydrolysis (KXXXFXSXXXXXS/TTN) motifs, the 3D pol (GLPSG and YGDD) motifs and the VP1 capsid (PPG) mo...
SummaryIn this study, the aetiological agent of diarrhea in Arabian thoroughbred foals housed in a stud farms and it's molecular characterization were reported. Out of sampled seven foals with diarrhoea, five were detected positive by RT-PCR depending on the amplification of the VP6 gene of rotavirus. Then the aetiological agent was characterized genetically by sequence analysis of the genome segments encoding VP6, VP7, VP4 and NSP4 of rotavirus. Findings revealed that the Turkish equine rotavirus circulating within this stud farm belongs to G3 and P[12] genotype with E2 NSP4 and I6VP6. This is the first study to report the G and P genotypes of equine group A rotaviruses in Turkey. Keywords: Rotavirus, Equine, Genotyping, Diarrhoea, Turkey Türkiye'de Bir Arap Atı İşletmesindeki Taylarda Rotavirus İshali Salgını ÖzetBu çalışmada, bir at yetiştiriciliği işletmesinde bulunan safkan arap taylarındaki ishal olgusunun etiyolojik ajanının tespiti ve moleküler karakterizasyonu bildirildi. İshal semptomlu yedi taydan sağlanan materyallerin beşinde RT-PCR tekniği ile rotavirus VP6 geni yönünden pozitiflik saptandı. Daha sonra enfeksiyona neden olan virusun VP6, VP7, VP4 ve NSP4 gen bölgelerini kodlayan gen bölgelerinin dizi analizi yapıldı. Elde edilen verilere dayanılarak söz konusu at yetiştiriciliği işletmesinde enfeksiyona neden olan ERV saha suşunun E2 NSP4 ve I6VP6 ile ilişkili G3P[12] genotipinde olduğu belirlendi. Bu çalışma, Türkiye'de atlardaki grup A rotavirusların G ve P genotiplerinin bildirildiği ilk çalışmadır.
Bu çalışmada, Kuzey Kıbrıs Türk Cumhuriyeti'nde (KKTC) yerleşik bir sığır yetiştiriciliği işletmesinde bulunan ishalli buzağılarda saptanan Grup A rotavirusun moleküler karakterizasyonu bildirildi. İshalli buzağılardan sağlanan dışkı örnekleri antijen ELISA ve Grup A rotavirus VP6 proteini kodlayan gen bölgeleri esas alınarak yapılan RT-PCR ile test edildi. Daha sonra enfeksiyona neden olan virusun VP4 ve VP7 proteinlerini kodlayan gen bölgeleri, generik ve tip spesifik primerler kullanılarak çoğaltıldı. Elde edilen verilere dayanılarak söz konusu işletmede bulunan buzağılarda enfeksiyona neden olan rotavirusun G6P[11] genotipe sahip olduğu belirlendi. Bu çalışma, KKTC'de buzağılarda Grup A rotavirusların G ve P genotiplerinin bildirildiği ilk çalışmadır. Anahtar sözcükler: Rotavirus, Buzağı, İshal, Genotip, KKTC The Molecular Characterization and Detection of Group A Rotavirus From Calves with Diarrhea in Turkish Republic of Northern Cyprus AbstractIn this study, the molecular characterization of the Group A rotavirus obtained from calves with diarrhea in a farm in TRNC was reported. Faces samples were detected as positive by ELISA and RT-PCR depending on the amplification of the VP6 gene of rotavirus. Then the aetiological agent was analyzed by RT-PCR with generic and type-spesific to the genome segments encoding VP4 and VP7 of rotavirus. Findings revealed that the rotavirus circulating within this farm belongs to G6 and P[11] genotype. This is the first study to report the G and P genotypes of Group A rotavirus from calves with diarrhea in TRNC.
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