Isolation and molecular characterization of anti biotic-resistant lactic acid bacteria from poultry and swine meat products
Isolation and molecular characterization of anti biotic-resistant lactic acid bacteria from poultry and swine meat products
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DOI：
10.4315/0362-028x-70.3.557
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发表时间：
2007-03-01
影响因子：
2
通讯作者：
Clementi, Francesca
中科院分区：
文献类型：
--
作者：
Aquilanti, Lucia;Garofalo, Cristiana;Clementi, Francesca
通过食物链从动物向人类转移对抗菌剂具有耐药性的微生物日益受到关注。为了确定非致病性微生物菌群对食物链中抗生素耐药性（AR）基因的发生和传播的贡献，从29个生的和加工的猪肉和鸡肉产品样品中分离出123种乳酸菌，这些样品先前对编码临床相关AR的一种或多种AR基因检测呈阳性：泰特（M）、泰特（O）、泰特（K）、tet（A）、tet（B）、tet（C）、aac（6 '）-Ie、aph（2”）-Ia、mecA和blaZ所有分离株最初都通过PCR检测其AR基因谱。通过分子鉴定，通过测定MIC进行分析，并进行遗传指纹分析，获得了59株携带泰特、tetB或blaZ基因的分离株。优势菌种为格氏乳球菌（28株），其次为植物乳杆菌（11株）和乳杆菌（11株）。唾液乳球菌（6株），而乳酸乳球菌乳酸亚种（6株）。乳酸乳杆菌（Lactobacillus johnsonii）、约氏乳杆菌（Lactobacillus johnsonii）、乳酸乳杆菌（L. n.罗伊特湖crispatus和L.在较低的频率下识别出短。泰特（M）和tetB（B）基因是最常见的基因。对18株泰特阳性（泰特（+））菌株进行多重耐药分析，结果表明：泰特（M）+tet（B）和泰特（K）+tet（B）是最常见的AR基因型。对三种选定菌株的泰特（M）开放阅读框进行部分测序，结果显示与先前在人类病原体（单核细胞增生李斯特菌和脑膜炎奈瑟菌）中发现的泰特（M）基因具有高度序列相似性（> 99%）。Southern杂交显示这些菌株含有携带泰特（M）的质粒。
The transfer via the food chain from animals to humans of microbes that are resistant to antimicrobial agents is of increasing concern. To determine the contributions of nonpathogenic microflora to the occurrence and spread of antibiotic resistance (AR) genes in the food chain, 123 lactic acid bacteria were isolated from 29 samples of raw and processed pork and chicken meat products that had previously tested positive for one or more AR genes that encode clinically relevant ARs: tet(M), tet(O), tet(K), erm(A), erm(B), erm(C), aac (6')-Ie aph (2")-Ia, mecA, and blaZ All of the isolates were initially tested for their AR gene profiles by PCR. The 59 isolates carrying a tet, erm, or blaZ gene were taken through molecular identification, analyzed by determination of the MIC, and subjected to genetic fingerprinting. Lactococcus garvieae was the predominant species (28 isolates), followed by Lactobacillus plantarum (11 isolates) and L. salivarius (6 isolates), whereas Lactococcus lactis subsp. lactis, Lactobacillus johnsonii, L. reuteri, L. crispatus, and L. brevis were identified at lower frequencies. The tet(M) and erm(B) genes were the most frequently detected. Assessment of multiple resistances in 18 tet positive (tet(+)) isolates revealed that tet(M) plus erm(B) and tet(K) plus erm(B) were the most frequent AR gene patterns. Partial sequencing of the tet(M) open reading frame of three selected strains showed high sequence similarities (> 99%) with tet(M) genes previously found in human pathogens (Listeria monocytogenes and Neisseria meningitidis). Southern hybridization with plasmid profiles revealed these strains contained tet(M)-carrying plasmids.