Molecular characteristics and drug resistance of Aeromonas from environmental Sources in Ma'anshan, Anhui
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摘要:
目的了解2017年安徽省马鞍山市环境来源气单胞菌的流行特征、毒力基因特征及耐药状况。 方法2017年对来自马鞍山市监测的海水产品、包装食品、生活用水、游泳池水样、食物中毒样本等进行气单胞菌分离检测,采用管家基因rpoD序列分析进行种水平鉴定,PCR检测气单胞菌6种毒力基因(Fla、Elastase、hlyA、ast、act和ascF-G)、脉冲场凝胶电泳(PFGE)分型分析分子型别,药敏试验检测药物敏感性。 结果共检测样品869份,气单胞菌检出率为7.6%(66/869),检出率最高的是农村生活用水、食物中毒样品和海水产品。 66株气单胞菌分为7个种,最多的种是维氏气单胞菌(36.4%,24/869)、嗜水气单胞菌(19.7%,13/869)和简氏气单胞菌(19.7%,13/869)。 64株气单胞菌携带至少1种毒力基因,Fla和act总检出率最高(53.1%),Fla在豚鼠气单胞菌中检出率最高(83.3%),act在嗜水气单胞菌检出率最高(69.2%)。 66株气单胞菌分为64个PFGE带型,表现为多样性。 药敏实验显示,气单胞菌对头孢替坦等7种抗生素100%敏感,对头孢唑啉等5种抗生素部分敏感(耐药率1.5% ~ 21.2%),而对氨苄西林、氨苄西林/舒巴坦的耐药率均高达90.0%。 结论2017年马鞍山地区气单胞菌污染较为严重,携带较高比例的多种毒力基因,并且对多种抗生素耐药性较高,应该加强对气单胞菌及其耐药性的监测。 Abstract:ObjectiveTo understand the epidemiological characteristics, virulence gene characteristics and drug resistance of Aeromonas from environmental sources in Ma’anshan of Anhui province. MethodsFrom January to December, 2017, Aeromonas strains isolated from seawater products, packaged foods, domestic water, swimming pool water and food poisoning samples in Ma'anshan were detected. The species of Aeromonas were identified through housekeeping gene rpoD sequence analysis. PCR was used to detect six virulence genes (Fla, Elastase, hlyA, ast, act and ascF-G), the molecular types of the strains were detected by using pulsed-field gel electrophoresis (PFGE), and drug susceptibility test of the strains was also conducted. ResultsA total of 66 Aeromonas strains were isolated from 869 samples (7.6%), and the isolated rates were higher in rural domestic water, food poisoning samples and seafood products respectively. Sixty-six strains of Aeromonas belonged to seven species, mainly including A. veronii (24, 36.4%), A. hydrophila (13, 19.7%) and A. jandaei (13, 19.7%) respectively. 64 strains of Aeromonas (97.0%) carried one or more virulence genes, with the highest detection rate of Fla and act (53.1%), Fla was mainly detected in A. caviae (83.3%) and act was mainly detected in A. hydrophila (69.2%). Sixty six strains of Aeromonas were divided into 64 PFGE types, indicating the diversity of molecular biotypes. Drug susceptibility test showed that Aeromonas was sensitive to seven antibiotics including cefotetan, and partially sensitive to five antibiotics (1.5%–21.2%) such as cefazolin, and highly resistant to ampicillin and ampicillin/sulbactam (90.0%). ConclusionThe contamination of Aeromonas, which carries multiple virulence genes and has high resistances to several antibiotics, is severe in Ma'anshan. It is necessary to strengthen the surveillance for Aeromonas and its drug resistance. -
Key words:
- Aeromonas /
- Molecular characteristic /
- Drug sensitivity
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表 1 气单胞菌毒力基因检测引物
Table 1. Primers used for detection of virulence genes of Aeromonas
毒力
基因GenBank
序列号引物序列(5′ ~ 3′) 产物长
度(bp)Fla FlaF TCCAACCGTYTGACCTC 608 FlaR GMYTGGTTGCRATGGT Elastase ElaF ACACGGTCAAGGAGATCAAC 513 ElaR CGCTGGTGTTGGCCAGCAGG hlyA hlyAF GGCCGGTGGCCCGAAGATACGGG 597 hlyAR GGCGGCGCCGGACGAGACGGG ast astF TCTCCATGCTTCCCTTCCACT 260 astR GTGTAGGGATTGAAGAAGCCG act actF AGAAGGTGACCACCAAGAACA 232 actR AACTGACATCGGCCTTGAACTC Ascf-G ASCF-GF ATGAGGTCATCTGCTCGCGC 900 ASCF-GR GGAGCACAACCATGGCTGAT 表 2 不同样本中气单胞菌检出情况
Table 2. Detection of Aeromonas in different samples
样品名称 样品数(份) 分离阳性数(份) 阳性率(%) 农村生活用水 127 35 27.6 食物中毒样品 82 10 12.2 海水产品 100 9 9.0 游泳池水 121 7 5.8 城区生活用水 278 5 1.8 包装食品 161 0 0.0 合计 869 66 7.6 表 3 不同样品中气单胞菌的种类分布
Table 3. Species distribution of Aeromonas in different samples
样品名称 维氏气单胞菌(株) 嗜水气单胞菌(株) 简氏气单胞菌(株) 水生气单胞菌(株) 豚鼠气单胞菌(株) 疱氏气单胞菌(株) 舒氏气单胞菌(株) 总计(株) 农村生活用水 18 4 9 3 0 0 1 35 食物中毒样品 3 1 2 0 4 0 0 10 海水产品 0 2 0 5 2 0 0 9 游泳池水 1 5 1 0 0 0 0 7 城区生活用水 2 1 1 0 0 1 0 5 合计 24 13 13 8 6 1 1 66 表 4 66株气单胞菌毒力基因检测情况
Table 4. Detections of virulence genes of Aeromonas strains
毒力
基因维氏气单胞菌(n=24) 嗜水气单胞菌(n=13) 简氏气单胞菌(n=13) 水生气单胞菌(n=8) 豚鼠气单胞菌(n=6) 合计 检出数(株) 检出率(%) 检出数(株) 检出率(%) 检出数(株) 检出率(%) 检出数(株) 检出率(%) 检出数(株) 检出率(%) 检出数(株) 检出率(%) Fla 13 54.2 6 46.2 5 38.5 5 62.5 5 83.3 34 53.1 act 12 50.0 9 69.2 6 46.2 3 37.5 4 66.7 34 53.1 Elastase 6 25.0 8 61.5 7 53.8 4 50.0 3 50.0 28 43.8 hlyA 8 33.3 6 46.2 4 30.8 4 50.0 2 33.3 24 37.5 Ast 9 37.5 6 46.3 4 30.8 3 37.5 3 50.0 23 35.9 Ascf-G 6 25.0 6 46.4 3 23.1 2 25.0 2 33.3 18 28.1 表 5 66株气单胞菌菌的药敏结果情况
Table 5. Drug susceptibility results of Aeromonas strains
药物名称 敏感 中度敏感 耐药 菌株数量(株) 敏感率(%) 菌株数量(株) 敏感率(%) 菌株数量(株) 耐药率(%) 氨苄西林 1 1.5 3 4.5 62 94.0 氨苄西林/舒巴坦 3 4.5 3 4.5 60 90.9 头孢唑啉 33 50.0 18 27.3 15 22.7 头孢替坦 66 100.0 0 0.0 0 0.0 头孢他啶 64 97.0 0 0.0 2 3.0 头孢曲松 65 98.5 0 0.0 1 1.5 氨曲南 64 96.7 0 0.0 2 3.3 亚胺培南 66 100.0 0 0.0 0 0.0 阿米卡星 65 98.5 1 1.5 0 0.0 庆大霉素 66 100.0 0 0.0 0 0.0 妥布霉素 65 98.5 1 1.5 0 0.0 环丙沙星 66 100.0 0 0.0 0 0.0 左旋氧氟沙星 66 100.0 0 0.0 0 0.0 呋喃妥因 66 100.0 0 0.0 0 0.0 甲氨苄啶/磺胺甲恶唑 65 98.5 0 0.0 1 1.5 哌拉西林/他唑巴坦 66 100.0 0 0.0 0 0.0 -
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2019-0151 马鞍山地区环境来源气单胞菌的分子特征和耐药分析.docx
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