Hu B, Kou Z Q, Shao C C, Yin H Y, Liu Z D, Xu X H, Fang M, Chen B L, Wei C Y, Li G F, Bi Z W
Shandong Provincial Center for Disease Control and Prevention, Ji'nan, Shandong 250014, China.
Zhonghua Yu Fang Yi Xue Za Zhi. 2018 Mar 6;52(3):271-276. doi: 10.3760/cma.j.issn.0253-9624.2018.03.010.
To understand the infection status, characteristics and drug resistance of non-O157 Shiga toxin-producing (STEC) in animal feces in Shandong Province. From 2015 to 2016, convient sampling method was used to collect 1 022 fresh feces of animals in Weishan county and Laizhou city, and 24 non-O157 STEC were isolated. The serotypes of non-O157 STEC strains were confirmed through serum agglutination test. The susceptibility was explored through the antimicrobial sensitivity experiments. ESBLs activity was confirmed by double-disc diffusion. PCR method was used to detect the resistance genes. PFGE typing was operated to assess the relatedness and variability of the strains. The multi-locus sequence typing (MLST) was adopted to get the allelic profile and ST sequence of strains. Analysis was made on the evolutionary relationship between different ST groups was made through CLC Sequence Viewer and Counting Express. A total of 24 non-O157 STEC were isolated from animal feces. 23 strains were from pig feces, and 1 strain was from cow feces, and the serotypes were more dispersed. All of the 24 strains carried s2 genes. The highest resistance rate was sulfamethoxazole(22 strains), the mount of cotrimoxazole and nalidixic acid was 18 strains, chloramphenicol was 13 strains, tetracycline was 19, and there was a phenomenon of multiple drug resistance. The drug resistance spectrum was sulfamethoxazole tetracycline-compound novammin-naphthidine-chloramphenicol. All strains were sensitive to cefepime and imipenem. The ESBLs confirmatory test showed that 4 strains of non O157 STEC produced beta lactamase. PCR detected 7 resistance genes, and 4 tetracycline resistance genes () were detected. The beta lactamase resistance genes (-1, -, ) were all negative. 24 strains were divided into 15 PFGE types, and their clustering results were more dispersed and no dominant PFGE type. There were 11 kinds of MLST types, most of them are ST540 and ST5133 types, each of which was 4 strains, and clustered into 1 MLST genomes. The serotypes of non-O157 STEC in animal feces O157 STEC were dispersed, and the resistant rate to common antibiotic was high. MLST typing results presents obvious polymorphism. Surveillance and manage ment of these strains should be strengthened.
为了解山东省动物粪便中非O157产志贺毒素大肠杆菌(STEC)的感染状况、特征及耐药性。2015年至2016年,采用方便抽样法收集微山县和莱州市1022份动物新鲜粪便,分离出24株非O157 STEC。通过血清凝集试验确定非O157 STEC菌株的血清型。通过药敏试验探索其药敏性。采用双纸片扩散法确认ESBLs活性。用PCR方法检测耐药基因。采用PFGE分型评估菌株的相关性和变异性。采用多位点序列分型(MLST)获取菌株的等位基因谱和ST序列。通过CLC Sequence Viewer和Counting Express对不同ST组之间的进化关系进行分析。从动物粪便中共分离出24株非O157 STEC。23株来自猪粪便,1株来自牛粪便,血清型较为分散。24株均携带s2基因。耐药率最高的是磺胺甲恶唑(22株),复方新诺明和萘啶酸均为18株,氯霉素为13株,四环素为19株,存在多重耐药现象。耐药谱为磺胺甲恶唑-四环素-复方新诺明-萘啶酸-氯霉素。所有菌株对头孢吡肟和亚胺培南敏感。ESBLs确证试验显示4株非O157 STEC产β-内酰胺酶。PCR检测到7种耐药基因,其中4种四环素耐药基因()被检测到。β-内酰胺酶耐药基因(-1、-、)均为阴性。24株分为15种PFGE型,其聚类结果较为分散,无优势PFGE型。有11种MLST型,其中多数为ST540和ST5133型,各有4株,聚为1个MLST基因组。动物粪便中非O157 STEC的血清型分散,对常用抗生素耐药率高。MLST分型结果呈现明显多态性。应加强对这些菌株的监测与管理。