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从食品洗出液中结合非靶标微生物到抗沙门氏菌和抗大肠杆菌 O157 免疫磁珠:背景真细菌最可能的组成。

Binding of nontarget microorganisms from food washes to anti-Salmonella and anti-E. coli O157 immunomagnetic beads: most probable composition of background Eubacteria.

机构信息

United States Department of Agriculture, Microbial Biophysics and Residue Chemistry, Eastern Regional Research Center, 600 E. Mermaid Lane, Wynmoor, PA 19038, USA.

出版信息

Anal Bioanal Chem. 2008 May;391(2):525-36. doi: 10.1007/s00216-008-1959-2. Epub 2008 Mar 3.

Abstract

We present herein the composition of bacterial communities occurring in ground chicken and the changes which arise in these populations based upon nonselective partitioning by commercially-available Dynal anti-Salmonella and anti-E. coli O157 immunomagnetic beads (IMB). Our enumeration and colony selection protocol was based upon a 6 × 6 drop plate method (n = 18 for each 25-g sub-sampling) using a dilution which resulted in ca. 4-8 colonies per drop. An average of 82 ± 13 colonies were selected from three 25-g ground chicken subsamplings per batch, each of which was repeated seasonally for one year. DNA was extracted from each colony and the composition of Eubacteria in each of these harvests was determined by sequence-based identification of 16S rDNA amplicons. The Gram-positive bacteria Brochothrix thermosphacta and Carnobacterium maltaromticum were the most commonly found organisms in both the total chicken wash (PBS) and in the IMB-bound (PBS-washed) fractions. The remaining background organisms which also adhered to varying degrees to commercial IMBs were: Pseudomonas oleovorans, Acinetobacter lwoffi, Serratia spp., and one Rahnella spp. A large number of the organisms were also cladistically evaluated based on rDNA basepair disparities: all Brochothrices were monophyletic; twelve different Pseudomonads were found along with eight Carnobacteria, seven Acinetobacteres, four Serratiae, and two Rahnellae. Carnobacterium alone showed an IMB-based concentration enhancement (ca. two to sixfold).

摘要

我们在此介绍了在鸡肉碎中出现的细菌群落的组成,以及基于商业上可用的 Dynal 抗沙门氏菌和抗大肠杆菌 O157 免疫磁珠(IMB)的非选择性分离,这些菌群会发生怎样的变化。我们的计数和菌落选择方案是基于 6×6 滴板法(每个 25-g 亚采样重复 18 次),稀释度导致每个滴约有 4-8 个菌落。每个批次从三个 25-g 的鸡肉碎亚采样中平均选择了 82±13 个菌落,每个批次在一年内重复进行季节性实验。从每个菌落中提取 DNA,并通过对这些收获物中的 16S rDNA 扩增子进行基于序列的鉴定来确定真细菌的组成。革兰氏阳性菌布罗特氏菌(Brochothrix thermosphacta)和麦芽糖球菌(Carnobacterium maltaromticum)是在总鸡肉冲洗液(PBS)和 IMB 结合(PBS 冲洗)部分中最常见的细菌。其余附着在不同程度的商业 IMB 上的背景细菌还有:食油假单胞菌(Pseudomonas oleovorans)、洛菲不动杆菌(Acinetobacter lwoffi)、沙雷氏菌属(Serratia spp.)和一个雷氏菌属(Rahnella spp.)。大量细菌也基于 rDNA 碱基差异进行了系统发育评估:所有布罗特氏菌都是单系的;发现了 12 种不同的假单胞菌,还有 8 种麦芽糖球菌、7 种不动杆菌、4 种沙雷氏菌和 2 种雷氏菌。仅麦芽糖球菌表现出基于 IMB 的浓度增强(约 2-6 倍)。

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