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在PALCAM培养基和Rapid' L. mono培养基上,低毒力单核细胞增生李斯特菌菌株的分离频率低于强毒力菌株。

Hypovirulent Listeria monocytogenes strains are less frequently recovered than virulent strains on PALCAM and Rapid' L. mono media.

作者信息

Gracieux Patrice, Roche Sylvie M, Pardon Pierre, Velge Philippe

机构信息

Institut National de la Recherche Agronomique, Laboratoire de Pathologie Infectieuse et Immunologie, 37380 Nouzilly, France.

出版信息

Int J Food Microbiol. 2003 Jun 15;83(2):133-45. doi: 10.1016/s0168-1605(02)00321-5.

Abstract

Several selective media have been developed to detect Listeria monocytogenes contaminated foodstuffs. Polymyxin-acriflavine-LiCl-ceftazidime-aesculin-mannitol (PALCAM) and Oxford media, required for the EN ISO method 11 290-1, are used for the detection of Listeria spp. in 2 days based on the expression of esculinase activity. Selective agar media such as Rapid' L. mono and Agar Listeria according to Ottaviani and Agosti (ALOA), based on the activity of phosphatidylinositol phospholipase C (PI-PLC) that allows the specific detection of L. monocytogenes in 2 days, are also used. However, no medium can assess the level of virulence of L. monocytogenes strains. Using a plaque-forming assay followed by subcutaneous footpad inoculation in mice, 15 virulent, 8 hypovirulent and 17 avirulent strains were discriminated among L. monocytogenes strains mainly originating from food (36/40). Their growth was tested on the four selective media. After 2 days, the number of colony forming units (cfu) of all the virulent strains was significantly superior to the number obtained with avirulent strains on all the four media tested, and superior to the number obtained with hypovirulent strains on PALCAM and Oxford media. These results showed a relationship between the level of virulence of L. monocytogenes strains and their growth on the selective agar media tested. Moreover, 1 out of 8 hypovirulent and 5 out of 17 avirulent strains did not grow on Rapid' L. mono medium, and 1 hypovirulent and 8 avirulent strains grew but did not express PI-PLC activity during the 7 days of incubation. The lack of detection of PI-PLC activity on Rapid' L. mono was not related to a gene mutation since these strains expressed enzymatic activity on ALOA medium, which detected up to 92% of the hypo- and avirulent strains. In contrast, some of these strains without growth or enzymatic activity expression would not be detected with PALCAM and Rapid' L. mono in foodstuffs on the second day.

摘要

已经开发出几种选择性培养基来检测被单核细胞增生李斯特菌污染的食品。EN ISO 11290-1方法所要求的多粘菌素-吖啶黄素-氯化锂-头孢他啶-七叶苷-甘露醇(PALCAM)培养基和牛津培养基,用于基于七叶苷酶活性检测李斯特菌属,在2天内完成检测。还有一些选择性琼脂培养基,如根据奥塔维亚尼和阿戈斯蒂(Ottaviani and Agosti)方法的快速单核细胞增生李斯特菌培养基(Rapid' L. mono)和阿戈斯蒂李斯特菌琼脂培养基(ALOA),基于磷脂酰肌醇磷脂酶C(PI-PLC)的活性,可在2天内特异性检测单核细胞增生李斯特菌。然而,没有一种培养基能够评估单核细胞增生李斯特菌菌株的毒力水平。通过噬斑形成试验,然后在小鼠皮下足垫接种,在主要源自食品的(36/40)单核细胞增生李斯特菌菌株中区分出15株强毒株、8株弱毒株和17株无毒株。在四种选择性培养基上测试了它们的生长情况。2天后,在所有四种测试培养基上,所有强毒株的菌落形成单位(cfu)数量显著高于无毒株,并且在PALCAM和牛津培养基上高于弱毒株。这些结果表明单核细胞增生李斯特菌菌株的毒力水平与其在测试的选择性琼脂培养基上的生长之间存在关联。此外,8株弱毒株中有1株、17株无毒株中有5株在快速单核细胞增生李斯特菌培养基上不生长,1株弱毒株和8株无毒株生长,但在7天的培养期间未表达PI-PLC活性。在快速单核细胞增生李斯特菌培养基上未检测到PI-PLC活性与基因突变无关,因为这些菌株在阿戈斯蒂李斯特菌琼脂培养基上表达酶活性,该培养基可检测高达92%的弱毒和无毒菌株。相比之下,这些没有生长或酶活性表达的菌株中的一些在第二天用PALCAM和快速单核细胞增生李斯特菌培养基在食品中无法检测到。

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