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Diarrhea outbreaks in suckling piglets due to rotavirus group C single and mixed (rotavirus groups A and B) infections

机译:轮状病毒C组单次和混合感染(轮状病毒A和B组)引起的乳猪腹泻暴发

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Porcine group A rotavirus (PoRVA) is a major cause of neonatal diarrhea in suckling and recently weaned piglets worldwide. The involvement of non-group A rotavirus in cases of neonatal diarrhea in piglets are sporadic. In Brazil there are no reports of the porcine rotavirus group C (PoRVC) as etiologic agent of the diarrhea outbreaks in piglets. The aim of this study was to describe the identification of rotavirus group C in single and in mixed infection with rotavirus groups A and B in three neonatal diarrhea outbreaks in suckling (<= 21-day-old) piglets, with 70% to 80% and 20% to 25% of morbidity and lethality rates, respectively, in three pig herds located in the state of Santa Catarina, Brazil. The diagnosis of PoRV in the diarrheic fecal samples was performed using polyacrylamide gel electrophoresis (PAGE) to identify the presence of porcine rotavirus groups A, B (PoRVB), and C, and by RT-PCR (PoRVA and PoRVC) and semi-nested (SN)-PCR (PoRVB) to partially amplify the VP4 (VP8*)-VP7, NSP2, and VP6 genes of PoRVA, PoRVB, and PoRVC, respectively. One RTPCR (PoRVA and PoRVC) and SN-PCR (PoRVB) product of each group of rotavirus of each diarrhea outbreak was submitted to nucleotide (nt) sequence analysis. Based on the PAGE technique, 4 (25%) and 1 (6.25%) of the 16 diarrheic fecal samples evaluated in the first outbreak presented PoRVA and PoRVC electropherotype, respectively, and 11 (68.75%) were negative. In the second outbreak, 3 (42.85%) of the 7 fecal samples evaluated presented PoRVA electropherotype, and in 3 (42.85%) and in 1 (14.3%) fecal samples were detected inconclusive and negative results, respectively. Three (30%) of the 10 fecal samples of the third outbreak presented PoRVC electropherotype; 5 (50%) and 2 (20%) samples showed negative and inconclusive results, respectively. Based on the RT-PCR and SN-PCR assays in the first neonatal diarrhea outbreak, PoRVC was detected in 13 (81.2%) of the 16 diarrheic fecal samples evaluated. PoRVC single infection was identified in 4 (25%) of these samples and mixed infections with PoRVA and PoRVB in 9 (56.2%) fecal samples. All of the seven diarrheic fecal samples evaluated from the second neonatal diarrhea outbreak were positive for PoRVC, whereas its mixed infection with other PoRV groups was detected in 4 (57.2%) samples. In the third outbreak, PoRVC in single infection was detected in all of the 10 diarrheic fecal samples analyzed. In the nt sequence analysis, the PoRVA strains of the first and second outbreaks demonstrated higher nt identity with G4P[6] and G9P[23] genotypes, respectively. The PoRVB strains (first and second outbreaks) and the PoRVC strains (first, second, and third outbreaks) showed higher nt identity and clustered in the phylogenetic tree with PoRVB and PoRVC strains that belong to the N4 and 11 genotypes, respectively. This is the first description in Brazil of the involvement of PoRVC in the etiology of diarrhea outbreaks in suckling piglets. The results of this study demonstrated that PoRVC, in both single and mixed infections, is an important enteropathogen involved in neonatal diarrhea outbreaks in piglets and that the use of more sensitive diagnostic techniques allows the identification of mixed infections involving two or even three groups of PoRV, which may be more common than previously reported.
机译:猪A型轮状病毒(PoRVA)是全球范围内哺乳和最近断奶的仔猪新生儿腹泻的主要原因。仔猪新生儿腹泻病例中非A型轮状病毒的感染是零星的。在巴西,没有关于猪轮状病毒C组(PoRVC)作为仔猪腹泻暴发病因的报道。这项研究的目的是描述在乳猪(<= 21日龄)的三例新生儿腹泻暴发中,轮状病毒C组在单次感染和混合感染中轮状病毒A和B组的鉴定。分别位于巴西圣卡塔琳娜州的三只猪群的发病率和致死率分别为20%至25%。使用聚丙烯酰胺凝胶电泳(PAGE)对猪轮状病毒A,B(PoRVB)和C组进行鉴定,并通过RT-PCR(PoRVA和PoRVC)进行半巢式诊断,以诊断腹泻性粪便中的PoRV。 (SN)-PCR(PoRVB)分别部分扩增PoRVA,PoRVB和PoRVC的VP4(VP8 *)-VP7,NSP2和VP6基因。将每个腹泻爆发的每组轮状病毒的一个RTPCR(PoRVA和PoRVC)和SN-PCR(PoRVB)产物进行核苷酸(nt)序列分析。根据PAGE技术,在第一次暴发中评估的16个腹泻粪便样品中,分别有4个(25%)和1个(6.25%)呈现PoRVA和PoRVC电泳型,其中11个(68.75%)呈阴性。在第二次暴发中,所评估的7个粪便样本中有3个(42.85%)呈现PoRVA电泳型,分别在3个(42.85%)和1个(14.3%)粪便样本中检测出结论性和阴性结果。在第三次暴发的10个粪便样本中,有3个(30%)表现出PoRVC电泳图型; 5(50%)和2(20%)样品分别显示阴性和不确定的结果。根据首次新生儿腹泻暴发中的RT-PCR和SN-PCR分析,在评估的16个腹泻粪便样本中有13个(占81.2%)检测到PoRVC。在这些样本中有4个(25%)鉴定出PoRVC单一感染,在9个样本(56.2%)的粪便中鉴定出PoRVA和PoRVB混合感染。从第二次新生儿腹泻暴发中评估的所有七个腹泻粪便样本均对PoRVC呈阳性,而在4个样本(57.2%)中检测到其与其他PoRV组的混合感染。在第三次爆发中,在所分析的所有10个腹泻粪便样本中均检测到了单一感染的PoRVC。在nt序列分析中,第一次和第二次暴发的PoRVA株分别显示出与G4P [6]和G9P [23]基因型更高的nt同一性。 PoRVB菌株(第一次和第二次暴发)和PoRVC菌株(第一次,第二次和第三次暴发)显示出较高的nt同一性,并在系统发育树中与分别属于N4和11基因型的PoRVB和PoRVC菌株聚集在一起。这是巴西对PoRVC参与乳猪腹泻暴发病因的首次描述。这项研究的结果表明,无论是单一感染还是混合感染,PoRVC都是仔猪新生儿腹泻暴发的重要肠病原体,而且使用更灵敏的诊断技术可以鉴定涉及两组或什至三组PoRV的混合感染,这可能比以前报道的更为常见。

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