Ranucci David, Battisti Elena, Veronesi Fabrizia, Diaferia Manuela, Morganti Giulia, Branciari Raffaella, Ferroglio Ezio, Valiani Andrea, Chiesa Francesco
Department of Veterinary Medicine, University of Perugia, San Costanzo Street 4, 06126 Perugia, Italy.
Department of Veterinary Science, University of Turin, Paolo Braccini Square 2, 10095 Grugliasco (TO), Italy.
Microorganisms. 2020 Jan 20;8(1):143. doi: 10.3390/microorganisms8010143.
The presence of viable was investigated in artisanal cheeses made from milk of naturally infected ewes. Ewe milk was analyzed beforehand for the presence and vitality of by loop-mediated isothermal amplification (LAMP) and reverse-transcriptase PCR (RT-PCR), respectively. Cheeses were prepared from raw milk following a traditional cheesemaking process. The cheese obtained from -positive milk was analyzed by LAMP to detect DNA-positive samples. RT-PCR was then carried out to assess the viability of the parasites in -positive milk samples and fresh cheese, after 5 and 15 days of ripening. Physical-chemical parameters of cheeses were also investigated. All cheese samples derived from -positive milk were positive according to LAMP, at both 5 and 15 days of ripening, while none of the samples were positive according to RT-PCR. Thus, while the presence of the parasite was demonstrated by the detection of specific DNA, the absence of detectable RNA supports the hypothesis that changes in the chemical and physical characteristics occurring during the cheesemaking process and ripening period, could be sufficient to inactivate viable in milk, minimizing the risk of human infection through consumption of raw sheep milk cheese.
对用自然感染母羊的乳汁制作的手工奶酪中是否存在活的(寄生虫名称未给出,原文此处缺失关键信息)进行了调查。分别通过环介导等温扩增法(LAMP)和逆转录聚合酶链反应(RT-PCR)对母羊乳汁进行预先分析,以检测(寄生虫名称未给出,原文此处缺失关键信息)的存在及活力。按照传统奶酪制作工艺用生乳制备奶酪。对从(寄生虫名称未给出,原文此处缺失关键信息)阳性乳汁获得的奶酪通过LAMP进行分析,以检测(寄生虫名称未给出,原文此处缺失关键信息)DNA阳性样本。然后在成熟5天和15天后,进行RT-PCR以评估(寄生虫名称未给出,原文此处缺失关键信息)阳性乳汁样本和新鲜奶酪中寄生虫的活力。还对奶酪的理化参数进行了研究。根据LAMP,所有源自(寄生虫名称未给出,原文此处缺失关键信息)阳性乳汁的奶酪样本在成熟5天和15天时均为阳性,而根据RT-PCR,所有样本均为阴性。因此,虽然通过检测特定DNA证明了寄生虫的存在,但未检测到(寄生虫名称未给出,原文此处缺失关键信息)RNA支持这样的假设,即在奶酪制作过程和成熟期发生的化学和物理特性变化可能足以使乳汁中的活(寄生虫名称未给出,原文此处缺失关键信息)失活,从而将通过食用生羊奶奶酪导致人类感染的风险降至最低。