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比较 7 种培养方法分离禽粪便中肠炎沙门氏菌和鼠伤寒沙门氏菌。

Comparison of 7 culture methods for Salmonella serovar Enteritidis and Salmonella serovar Typhimurium isolation in poultry feces.

机构信息

Instituto Nacional de Tecnología Agropecuaria EEA Concepción del Uruguay, 3260, Concepción del Uruguay, Entre Ríos, Argentina.

Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Laboratorio de Sanidad Aviar, Instituto Nacional de Tecnología Agropecuaria EEA Concepción del Uruguay, 3260, Concepción del Uruguay, Entre Ríos, Argentina.

出版信息

Poult Sci. 2018 Nov 1;97(11):3826-3836. doi: 10.3382/ps/pey259.

Abstract

The present work compared 7 different culture methods and 3 selective-differential plating media for Salmonella ser. Enteritidis (SE) and S. ser. Typhimurium (ST) isolation using artificially contaminated poultry feces. The sensitivity (Se) and accuracy (AC) values increased when Modified Semisolid Rappaport Vassiliadis (MSRV) methods were used in place of the Tetrathionate (TT) or Tetrathionate Hajna broth (TTH) method in the enrichment step. However, there was no significant difference between the pre-enrichment incubation at 4 to 6 and 18 to 24 h for MSRV5 and MSRV24 methods, respectively. All Salmonella strains were recovered in the lowest dilutions tested for MSRV24 and 3 out of 4 for MSRV5 methods (2 to 10 cfu/25 g). The TT and TTH methods showed a detection limit between 2.2 × 101 and 1.0 × 106 cfu/25 g of fecal sample. The agreement was variable between the methods. However, there was a very good agreement between the MSRV5 and MSRV24 methods, and between tetrathionate direct (TTD, no pre-enrichment media used) and buffered peptone water 18 to 24 h Tetrathionate broth combination (TT24 method) for Salmonella strains. The 3 selective-differential plating media showed an agreement between fair and excellent. They performed a high Se and AC in the MSRV methods for Salmonella strains. There was a significant difference between center and periphery for MSRV methods, and there was a fair agreement between them for all strains. The MSRV methods are better than TT/TTH methods for the isolation of different strains of SE and ST in poultry fecal samples. The MSRV5 method can be used to reduce the time for the detection of SE and ST in these samples. Furthermore, a loopful of the periphery of the growth should be streaked onto differential-selective plating media, even in the absence of halo, to decrease the number of false negative results.

摘要

本研究比较了 7 种不同的培养方法和 3 种选择性差异平板培养基,用于从人工污染的家禽粪便中分离肠炎沙门氏菌(SE)和鼠伤寒沙门氏菌(ST)。在富集步骤中,用改良半固体雷帕氏培养基(MSRV)方法替代四硫酸盐(TT)或四硫酸盐-哈氏培养基(TTH)方法时,灵敏度(Se)和准确性(AC)值增加。然而,MSRV5 和 MSRV24 方法的预富集孵育时间分别为 4 至 6 小时和 18 至 24 小时,之间没有显著差异。所有沙门氏菌菌株均在 MSRV24 方法的最低稀释度下回收,4 株中有 3 株在 MSRV5 方法中回收(2 至 10 cfu/25 g)。TT 和 TTH 方法的检测限介于 2.2 × 101 和 1.0 × 106 cfu/25 g 粪便样本之间。方法之间的一致性各不相同。然而,MSRV5 和 MSRV24 方法之间、四硫酸盐直接法(TTD,不使用预富集培养基)和缓冲蛋白胨水 18 至 24 小时四硫酸盐肉汤组合法(TT24 方法)之间的一致性非常好。3 种选择性差异平板培养基之间的一致性为良好到极好。它们在 MSRV 方法中对沙门氏菌菌株具有高 Se 和 AC。MSRV 方法的中心和外周之间存在显著差异,所有菌株之间的一致性良好。MSRV 方法优于 TT/TTH 方法,可用于从家禽粪便样本中分离不同的 SE 和 ST 菌株。MSRV5 方法可用于缩短这些样本中 SE 和 ST 的检测时间。此外,即使没有晕环,也应该用一环外周的生长物划线接种到差异选择性平板培养基上,以减少假阴性结果的数量。

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