越南猪副嗜血杆菌病毒的分离和特征描述

T. T. H. Trinh, V. T. Do, V. K. Do, H. Vu-Khac
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摘要

背景和目的:还没有研究成功分离出越南的副猪嗜血杆菌病毒。本研究旨在分离越南本土的副猪嗜血杆菌病毒株并确定其特征,以开发猪副猪嗜血杆菌病毒(PPV)疫苗。材料和方法:我们从越南 6 个省采集了血清和死胎样本,并使用聚合酶链反应鉴定了 PPV 阳性样本。使用在添加了 5%胎牛血清和 1%抗生素(青霉素-链霉素)的最低限度基本培养基中培养的 PK-15 细胞尝试分离 Parvovirus。细胞在 37°C 和 5% CO2 条件下培养。在白色初产母猪身上进行了毒力实验,通过血凝抑制(HI)滴度和胎儿病变来评估 PPV 株的毒力。结果我们分析了 360 份血清样本和 32 份死胎(肝脏和肺脏)样本,发现其中 32/392 份(8.2%)为 PPV 阳性,全部属于 PPV1 型。成功分离出 32 份 PPV 阳性样本,其 VP2 序列的一致性为 100%。系统发生树显示,它们与 Kresse 株系(1996 年分离自加拿大)和 PPV1-0225-L-SD 株系(2022 年分离自中国)关系密切。两个 PPV 分离株(东内的 VC5 和清化的 TX7)表现出较高的 50% 组织培养感染剂量滴度,被选中用于毒力实验。注射后第 21 天,HI 抗体滴度在 10log2 至 12log2 之间。第 90 天,71%-80% 的胎儿变成木乃伊。结论这项研究表明,越南的 PPV 感染率为 8.2%。32个分离株属于PPV1。根据后备母猪注射后的 HI 滴度结果,VC5 和 TX7 这两种 PPV 株系被确定为高致病性。VC5和TX7被确定为进一步研究PPV疫苗的良好候选株。关键词:分离、系统发生树、猪副嗜血杆菌、母猪、毒力。
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Isolation and characterization of porcine parvovirus in Vietnam
Background and Aim: No study has successfully isolated parvovirus in Vietnam. This study aimed to isolate and characterize parvovirus strains indigenous in Vietnam for vaccine development against porcine parvovirus (PPV). Materials and Methods: We collected serum and stillbirth samples from six provinces in Vietnam, and PPV-positive samples were identified using a polymerase chain reaction. Parvovirus isolation was attempted using the PK-15 cells maintained in a minimum essential medium supplemented with 5% fetal bovine serum and 1% antibiotics (Penicillin-streptomycin). The cells were incubated at 37°C with 5% CO2. Virulence experiments were conducted on white primiparous sows to evaluate the virulence of the PPV strain through hemagglutination inhibition (HI) titers and fetus lesions. Results: We analyzed 360 serum and 32 stillbirth (liver and lungs) samples, revealing that 32/392 (8.2% ) of them were PPV-positive, all belonging to PPV1. Thirty-two PPV-positive samples were successfully isolated, with 100% identity as VP2 sequences. The phylogenetic tree revealed a close relationship with the Kresse strain (isolated from Canada in 1996) and the PPV1-0225-L-SD strain (isolated from China in 2022). Two PPV isolates (VC5 from Dongnai and TX7 from Thanhhoa) that exhibited high 50% tissue culture infectious dose titers were selected for the virulence experiment. On day 21, after injection, the HI antibody titers ranged from 10log2 to 12log2. On day 90, 71%–80% of fetuses were mummified. Conclusion: This study showed that the PPV infection rate in Vietnam was 8.2%. Thirty-two isolates belonged to PPV1. Two PPV strains, VC5 and TX7, were determined to be highly virulent by the results of HI titers after injection into gilts. VC5 and TX7 were determined to be good candidates for further research on PPV vaccines. Keywords: isolation, phylogenetic tree, porcine parvovirus, sow, virulence.
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