Faculty of Veterinary Science, Department of Veterinary Microbiology, Chulalongkorn University, Bangkok, 10330, Thailand.
Probiotics Antimicrob Proteins. 2018 Jun;10(2):383-390. doi: 10.1007/s12602-017-9281-y.
Porcine epidemic diarrhea virus (PEDV) is a coronavirus which causes severe diarrhea and fatal dehydration in piglets. In general, probiotic supplements could enhance recovery and protect piglets against enteric pathogens. Seven local lactic acid bacteria (LAB), (Ent. faecium 79N and 40N, Lact. plantarum 22F, 25F and 31F, Ped. acidilactici 72N and Ped. pentosaceus 77F) from pig feces were well-characterized as high potential probiotics. Cell-free supernatants (CFS) and live LAB were evaluated for antiviral activities by co-incubation on Vero cells and challenged with a pandemic strain of PEDV isolated from pigs in Thailand. Cell survival and viral inhibition were determined by cytopathic effect (CPE) reduction assay and confirmed by immunofluorescence. At 1:16, CFS dilution (pH 6.3-6.8) showed no cytotoxicity in Vero cells and was therefore used as the dilution for antiviral assays. The diluted CFS of all Lact. plantarum showed the antiviral effect against PEDV; however, the same antiviral effect could not be observed in Ent. faecium and Pediococcus strains. In competitive experiment, only live Lact. plantarum 25F and Ped. pentosaceus 77F showed CPE reduction in the viral infected cells to <50% observed field area. This study concluded that the CFS of all tested lactobacilli, and live Lact. plantarum (22F and 25F) and Pediococcus strains 72N and 77F could reduce infectivity of the pandemic strain of PEDV from pigs in Thailand on the target Vero cells.
猪流行性腹泻病毒（PEDV）是一种冠状病毒，可导致仔猪严重腹泻和致命脱水。一般来说，益生菌补充剂可以增强恢复能力并保护仔猪免受肠道病原体的侵害。从猪粪便中分离出的 7 株本地乳酸菌（LAB）（肠球菌 79N 和 40N、植物乳杆菌 22F、25F 和 31F、粪肠球菌 72N 和戊糖片球菌 77F）被很好地鉴定为高潜力益生菌。通过与从泰国猪中分离出的大流行株 PEDV 在 Vero 细胞中共孵育并进行挑战，评估无细胞上清液（CFS）和活 LAB 的抗病毒活性。通过细胞病变效应（CPE）减少测定法和免疫荧光来确定细胞存活和病毒抑制。在 1:16 的 CFS 稀释度（pH 6.3-6.8）下，Vero 细胞无细胞毒性，因此用作抗病毒测定的稀释度。所有植物乳杆菌的稀释 CFS 均显示出对 PEDV 的抗病毒作用；然而，在肠球菌和肠球菌菌株中无法观察到相同的抗病毒作用。在竞争实验中，只有活的植物乳杆菌 25F 和戊糖片球菌 77F 显示出在感染病毒的细胞中 CPE 减少到<50%观察到的视野面积。本研究得出结论，所有测试的乳杆菌的 CFS，以及活的植物乳杆菌（22F 和 25F）和肠球菌菌株 72N 和 77F 可以降低来自泰国猪的大流行株 PEDV 对靶 Vero 细胞的感染力。
