猪源气单胞菌分离鉴定及耐药谱检测
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摘要
为调查气单胞菌在我国猪粪便中流行情况及耐药谱,2017年3～7月从河南省4个地区,共采集370份猪粪样品,通过缓冲液蛋白胨水增菌培养后,使用添加有氨苄青霉素的气单胞菌基础培养基选择培养,分离气单胞菌。可疑菌株通过形态特征、生化试验和16S rDNA序列比对进一步鉴定。微量稀释法测定分离菌株对17种抗菌药物耐药谱。结果显示,共分离到11株气单胞菌,分离率为2.97%,其中豚鼠气单胞菌5株(占45.45%),杀鲑气单胞菌2株(占18.18%),中间气单胞菌、嗜水气单胞菌、水生气单胞菌和贝斯特润气单胞菌各1株(各占9.09%);阿莫西林/克拉维酸菌耐药率为100%,磺胺甲恶唑、氟苯尼考耐药率81.82%,阿米卡星、美罗培南敏感率均为100%,除了其中1株气单胞菌,其他菌株对头孢噻呋、头孢曲松、庆大霉素、强力霉素、磷霉素、恩诺沙星均敏感,各菌株均呈多重耐药性。表明我国健康猪粪便中携带气单胞菌,如果造成水源或者食物污染,可能有引起人类或者动物感染的风险,应引起重视。
In order to assess the incidence and antimicrobial resistance profile of Aeromonas spp.isolated from pig feces,we collected 370 samples of pig feces from farms located at Hebi,Xinxiang,Shangqiu and Nanyang,Henan province for isolation of Aeromonas spp.using Aeromonas medium with ampicillin selective supplement after the enriched culture in buffered peptone water.Aeromonasisolates were confirmed by morphological characteristics,biochemical tests and 16 Sr DNA sequence alignment.Antibiogram of Aeromonas isolates to 17 antimicrobial agents was done using the microdilution methods.The results of the study indicated that 11 strains of Aeromonas were isolated,the faecal carriage rate of Aeromonas was2.97%.Identification of isolates revealed 6 phenotypic species with percentage distribution as follows:Aeromonas caviae 5(45.45 %),Aeromonas salmonsalis 2(18.18% %),Aeromonas media 1(9.09%),Aeromonas hydrophila 1(9.09%),Aeromonas aquatic 1(9.09%),and Aeromonas bestiarum 1(9.09%).All Aeromonas isolates in the study were absolutely resistant to amoxicillin/clavulanic acid,and susceptible to amikacin and meropenem,while the resistance rate of sulfamethoxazole or florfenicol was 81.82%.All Aer omonasisolates except one isolate were susceptible to ceftiofur,ceftriaxone,gentamicin,doxycycline,fosfomycin,and enrofloxacin.All Aeromonas isolates were multi-drug resistant.This study suggested that healthy pig feces in China may carry Aeromonas,of which some may possess virulence,and once these isolates pollute water or food/feed,they can cause the risk to infections of human or animals.
In order to assess the incidence and antimicrobial resistance profile of Aeromonas spp.isolated from pig feces,we collected 370 samples of pig feces from farms located at Hebi,Xinxiang,Shangqiu and Nanyang,Henan province for isolation of Aeromonas spp.using Aeromonas medium with ampicillin selective supplement after the enriched culture in buffered peptone water.Aeromonasisolates were confirmed by morphological characteristics,biochemical tests and 16 Sr DNA sequence alignment.Antibiogram of Aeromonas isolates to 17 antimicrobial agents was done using the microdilution methods.The results of the study indicated that 11 strains of Aeromonas were isolated,the faecal carriage rate of Aeromonas was2.97%.Identification of isolates revealed 6 phenotypic species with percentage distribution as follows:Aeromonas caviae 5(45.45 %),Aeromonas salmonsalis 2(18.18% %),Aeromonas media 1(9.09%),Aeromonas hydrophila 1(9.09%),Aeromonas aquatic 1(9.09%),and Aeromonas bestiarum 1(9.09%).All Aeromonas isolates in the study were absolutely resistant to amoxicillin/clavulanic acid,and susceptible to amikacin and meropenem,while the resistance rate of sulfamethoxazole or florfenicol was 81.82%.All Aer omonasisolates except one isolate were susceptible to ceftiofur,ceftriaxone,gentamicin,doxycycline,fosfomycin,and enrofloxacin.All Aeromonas isolates were multi-drug resistant.This study suggested that healthy pig feces in China may carry Aeromonas,of which some may possess virulence,and once these isolates pollute water or food/feed,they can cause the risk to infections of human or animals.
引文
[1] Janda J M,Abbott S L.The genus Aeromonas:Taxonomy,pathogenicity,and infection[J].Clin Microbiol Rev,2010,23(1):35-73.
[2] Kimura M,Araoka H,Yoneyama A.Aeromonas caviae is the most frequent pathogen amongst cases of Aeromonas bacteremia in Japan[J].Scand J Infect Dis,2013,45(4):304-309.
[3] Igbinosa I H,Igumbor E U,Aghdasi F,et al.Emerging Aeromonas species infections and their significance in public health[J].Sci World J,2012:1-13.DOI:10.1100/2012/625023.
[4] Pablos M,Huys G,Cnockaert M,et al.Identification and epidemiological relationships of Aeromonas isolates from patients with diarrhea,drinking water and foods[J].Int J Food Microbiol,2011,147(3):203-210.
[5] Evangelopoulou G,Filioussis G,Kritas S,et al.Isolation and antimicrobial testing of Aeromonas spp.,Citrobacter spp.,Cronobacter spp.,Enterobacter spp.,Escherichia spp.,Klebsiella spp.,and Trabulsiella spp.from the gallbladder of pigs[J].Polish J Microbiol,2015,64(2):185-188.
[6] Igbinosa I H,Igbinosa E O,Okoh A I.Antibiogram characterization and putative virulence genes in Aeromonas species isolated from pig fecal samples[J].Environ Sci Pollut Res Int,2016,23(12):12199-12205.
[7] Gray S J,Stickler D J.Some observations on the faecal carriage of mesophilic Aeromonas species in cows and pigs[J].Epidemiol Infect,1989,103(3):523-537.
[8] Fontes M C,Saavedra M J,Martins C,et al.Phylogenetic identification of Aeromonas from pigs slaughtered for consumption in slaughterhouses at the North of Portugal[J].Int J Food Microbiol,2011,146(2):118-222.
[9] 东秀珠,蔡妙英.常见细菌系统鉴定手册[M].北京:科学出版社,1999:117-118,370.
[10] Weisburg W G,Barns S M,Pelletier D A,et al.16S ribosomal DNA amplification for phylogenetic study[J].J Bacteriol,1991,173(2):697-703.
[11] 李莉,陈颖,张超,等.水产动物气单胞菌鉴定方法研究进展[J].水产科学,2015,34(2):128-132.
[12] Beaz-Hidalgo R,Latif-Eugenín F,Hossain M J,et al.Aeromonas aquatica sp.nov.,Aeromonas finlandiensis sp.nov.and Aeromonas lacus sp.nov.isolated from Finnish waters associated with cyanobacterial blooms[J].Syst Appl Microbiol,2015,38(3):161-168.
[13] Fontes M C,Saavedra M J,Martins C,et al.Phylogenetic identification of Aeromonas from pigs slaughtered for consumption in slaughterhouses at the North of Portugal[J].Int J Food Microbiol,2011,146 (2):118-122.
[14] 方海英,李医,陈晓,等.肠道外感染气单胞菌属的临床分布与耐药性分析[J].中华医院感染学杂志,2015,25(7):1454-1456.
[2] Kimura M,Araoka H,Yoneyama A.Aeromonas caviae is the most frequent pathogen amongst cases of Aeromonas bacteremia in Japan[J].Scand J Infect Dis,2013,45(4):304-309.
[3] Igbinosa I H,Igumbor E U,Aghdasi F,et al.Emerging Aeromonas species infections and their significance in public health[J].Sci World J,2012:1-13.DOI:10.1100/2012/625023.
[4] Pablos M,Huys G,Cnockaert M,et al.Identification and epidemiological relationships of Aeromonas isolates from patients with diarrhea,drinking water and foods[J].Int J Food Microbiol,2011,147(3):203-210.
[5] Evangelopoulou G,Filioussis G,Kritas S,et al.Isolation and antimicrobial testing of Aeromonas spp.,Citrobacter spp.,Cronobacter spp.,Enterobacter spp.,Escherichia spp.,Klebsiella spp.,and Trabulsiella spp.from the gallbladder of pigs[J].Polish J Microbiol,2015,64(2):185-188.
[6] Igbinosa I H,Igbinosa E O,Okoh A I.Antibiogram characterization and putative virulence genes in Aeromonas species isolated from pig fecal samples[J].Environ Sci Pollut Res Int,2016,23(12):12199-12205.
[7] Gray S J,Stickler D J.Some observations on the faecal carriage of mesophilic Aeromonas species in cows and pigs[J].Epidemiol Infect,1989,103(3):523-537.
[8] Fontes M C,Saavedra M J,Martins C,et al.Phylogenetic identification of Aeromonas from pigs slaughtered for consumption in slaughterhouses at the North of Portugal[J].Int J Food Microbiol,2011,146(2):118-222.
[9] 东秀珠,蔡妙英.常见细菌系统鉴定手册[M].北京:科学出版社,1999:117-118,370.
[10] Weisburg W G,Barns S M,Pelletier D A,et al.16S ribosomal DNA amplification for phylogenetic study[J].J Bacteriol,1991,173(2):697-703.
[11] 李莉,陈颖,张超,等.水产动物气单胞菌鉴定方法研究进展[J].水产科学,2015,34(2):128-132.
[12] Beaz-Hidalgo R,Latif-Eugenín F,Hossain M J,et al.Aeromonas aquatica sp.nov.,Aeromonas finlandiensis sp.nov.and Aeromonas lacus sp.nov.isolated from Finnish waters associated with cyanobacterial blooms[J].Syst Appl Microbiol,2015,38(3):161-168.
[13] Fontes M C,Saavedra M J,Martins C,et al.Phylogenetic identification of Aeromonas from pigs slaughtered for consumption in slaughterhouses at the North of Portugal[J].Int J Food Microbiol,2011,146 (2):118-122.
[14] 方海英,李医,陈晓,等.肠道外感染气单胞菌属的临床分布与耐药性分析[J].中华医院感染学杂志,2015,25(7):1454-1456.