疾病监测  2016, Vol. 31 Issue (3): 225-228

扩展功能

文章信息

王闻卿, 丁明, 苏靖华, 傅慧琴, 赵冰, 张勇琪, 黄红, 章红红, 朱林英, 朱渭萍, 傅益飞, 孙乔, 许学斌
WANG Wen-qing, DING Ming, SU Jing-hua, FU Hui-qin, ZHAO Bing, ZHANG Yong-qi, HUANG Hong, ZHANG Hong-hong, ZHU Lin-ying, ZHU Wei-ping, FU Yi-fei, SUN Qiao, XU Xue-bin
上海市浦东新区腹泻病例中气单胞菌的流行和耐药特征研究
Prevalence and antibiotic resistance of Aeromonasfrom diarrheal patients in Pudong New Area, Shanghai
疾病监测, 2016, 31(3): 225-228
Disease Surveillance, 2016, 31(3): 225-228
10.3784/j.issn.1003-9961.2016.03.011

文章历史

收稿日期: 2015-11-26
上海市浦东新区腹泻病例中气单胞菌的流行和耐药特征研究
王闻卿 , 丁明, 苏靖华, 傅慧琴, 赵冰, 张勇琪, 黄红, 章红红, 朱林英, 朱渭萍, 傅益飞, 孙乔, 许学斌    
上海市浦东新区疾病预防控制中心, 上海 200136
摘要: 目的 了解上海市浦东新区腹泻病例中气单胞菌的流行及抗生素耐药特征。方法 2014年,在辖区内11家临床机构采集腹泻病例样本进行包括气单胞菌在内的腹泻多病原菌的检测,应用改良K-B法检测气单胞菌分离菌株对12种抗生素纸片的耐药性。结果 基于腹泻病例的实验室多病原诊断从1846份腹泻样本中共确认73例气单胞菌感染病例:单一型感染42例(57.6%),其中嗜水气单胞菌(嗜水)8例、维隆气单胞菌温和生物变种(温和)26例、豚鼠气单胞菌(豚鼠)8例;混合型感染31例(42.4%),总感染阳性率为4.0%。感染优势菌型为温和(P<0.05),感染峰值为年度6-8月间(9.0%),病例的男女性别比差异无统计学意义,病例的年龄组以0~4岁和40~91岁人群居多。有65.8%(48/73)的气单胞菌对至少1种抗生素产生耐药性,以阿莫西林/克拉维酸(38.4%)和四环素(34.2%)的耐药率为最高,发现的3株多重耐药(MDR)株均为温和气单胞菌,其中1株对6种抗生素多重耐药(包括头孢噻肟、环丙沙星和左氧氟沙星)。结论 浦东新区和食源性感染性腹泻相关的气单胞菌病例存在较高的混合感染现象,发病有显著的季节特征,5岁以下幼儿和中老年人较为易感,温和气单胞菌是优势菌型,其对抗生素的多重耐药水平有升高趋势。
关键词: 气单胞菌    食源性感染性腹泻    混合感染    多重耐药    
Prevalence and antibiotic resistance of Aeromonasfrom diarrheal patients in Pudong New Area, Shanghai
WANG Wen-qing , DING Ming, SU Jing-hua, FU Hui-qin, ZHAO Bing, ZHANG Yong-qi, HUANG Hong, ZHANG Hong-hong, ZHU Lin-ying, ZHU Wei-ping, FU Yi-fei, SUN Qiao, XU Xue-bin    
Pudong New District Center for Disease Control and Prevention, Shanghai, Shanghai 200136, China
Corresponding author: WANG Wen-qing, Email: wwq95@163.com
This study was supported by Young Medical Talent Training Program of Health System of Pudong New Area, Shanghai (No. PWRq2014-26)
Abstract: Objective To investigate the distribution and antibiotic resistance of Aeromonas isolated from the patients with acute diarrhea in Pudong New Area, Shanghai. Methods In 2014, Eleven hospitals were selected as sentinels in Pudong. The stool samples of diarrheal patients in 11 sentinel hospitals in Pudong were collected for the detection of diarrheal pathogens, including Aeromonas. The isolates were further detected for the susceptibility to 12 antibiotics with K-B method. Results A total of 73 Aeromonas infections were detected in 1846 samples (4.0%), including 42 single infections(57.6%) 8 A. hydrophila infections, 26 A. veronii bv. sobria infections, 8 A. caviae infections, and 31 co-infections with other pathogens(42.4%). A. veronii bv. sobria was the predominant pathogen (P<0.05). The infection peak was during June-August (9.0%), the positive rates between males and females had no statistical significance, and the cases in age groups 0~4 years and 40-91 years accounted for 65.8%(48/73). Most Aeromonas strains were resistant to at least one antibiotic. The resistance rate to amoxicillin-clavulanic acid was highest(38.4%), followed by that to tetracycline(34.2%).Three MDR strains of A.veronii bv. sobria were detected, including 1 strain resistant to cefotaxime, ciprofloxacin, levofloxacin and other 3 antibiotics. Conclusion Aeromonas caused diarrhea had obviously seasonal/foodborne infection characteristics. High proportion of co-infections occurred. People aged<5 years and >40 years were susceptible. A.veronii bv.sobria was the predominant pathogen with increased MDR level.
Key words: Aeromonas    Foodborne infectious diarrhea    Co-infection    MDR    

气单胞菌在自然界(尤其是淡、海水)广泛存在,是评价环境卫生学的重要指标菌[1]。已知的常见菌型有嗜水气单胞菌(嗜水)、维隆气单胞菌温和生物变种(温和)和豚鼠气单胞菌(豚鼠),可引发人体的肠道内感染(腹泻)或肠道外感染(伤口感染、败血症、蜂窝组织炎、腹膜炎)等,以不同年龄组的轻或中度腹泻最常见[1]。欧美等发达国家将气单胞菌列为潜在的健康危害风险因素[2]。近年有关产超广谱β-内酰胺酶(ESBLs)气单胞菌的报道再度引发对全球耐药菌防控问题的关注和讨论[1]。我国尚未将气单胞菌纳入国家肠道传染病的常规监测和传报体系,2014年,笔者利用上海市浦东新区开展的基于实验室诊断的腹泻病例多病原菌的监测项目平台,研究气单胞菌腹泻病例的流行和耐药特征,为科学防控本地区的食源性感染性腹泻提供循证依据。

1 材料与方法 1.1 样本来源

将上海市浦东新区辖区内开设肠道门诊的11家医院作为监测点,对符合急性腹泻病例定义且自诉未服用抗生素的病例[3],在知情同意基础上采集粪便拭子置于运送管,当天送达浦东新区疾病预防控制中心的实验室检测。

1.2 病原菌

气单胞菌、沙门菌、志贺菌、5种致泻性大肠埃希菌(EAEC、EPEC、ETEC、EIEC、STEC/EHEC)、小肠结肠炎耶尔森菌、弯曲菌、霍乱弧菌、副溶血弧菌、拟态弧菌、河弧菌、类志贺邻单胞菌。检测的材料方法见文献[3]。

1.3 药敏实验

采用改良K-B法(纸片扩散法),测试气单胞菌对12种抗生素的耐药性,结果判定参照2010年版美国临床实验室标委会(CLSI)标准[4]。抗生素纸片:阿莫西林/克拉维酸(AMC)、头孢吡肟(FEP)、头孢噻肟(CTX)、头孢他啶(CAZ)、亚胺培南(IPM)、阿米卡星(AK)、庆大霉素(GN)、四环素(TE)、环丙沙星(CIP)、左氧氟沙星(LEV)、氯霉素(C)、复方新诺明(SXT)(Oxoid,英国)。

1.4 数据处理与分析

使用Excel 2007软件进行数据处理,SPSS 16.0软件进行统计学分析,率的比较使用χ2检验和Fishers确切概率法,P <0.05判定为差异有统计学意义。

2 结果 2.1 气单胞菌腹泻病例的流行特征

经实验室对1846份腹泻样本进行多病原菌诊断,共甄别73例气单胞菌腹泻病例:单一型感染42例(57.6%),其中嗜水8例、温和26例、豚鼠8例;混合型感染31例(42.4%),总感染(阳性)率为4.0%。气单胞菌腹泻病例在四季皆有分布,以夏季峰值最高(9.0%)(P <0.05);男性和女性病例的阳性率差异无统计学意义(P>0.05);病例分布的年龄组最高依次为0~4(4.8%)、40~59(4.2%)、60~91(4.2%)和20~39岁(3.5%),见表 1

表1 基于腹泻多病原菌诊断的气单胞菌腹泻病例的流行病学分布特征 Table 1 Distribution of Aeromonas infected patients based on multi-pathogen detection
流行特征病例分布
(%)
单一型气单胞菌病例混合型气单胞菌病例
合计(%)嗜水温和豚鼠合计(%)嗜水温和豚鼠
季节a
(月)
春(35)2.8(9/321)8(88.9)5211(11.1)100
夏(68)9.0(45/498)24(53.3)216621(46.7)4134
秋(911)2.6(16/615)7(43.8)1519(56.2)450
冬(122)0.7(3/421)3(100.0)0300(0)000
合计73428268319184
性别b3.9(36/925)20(55.6)312516(44.4)493
4.0(37/921)22(59.5)514315(40.5)591
合计73428268319184
年龄组c
(岁)
0~4.8(12/250)6(50.0)2226(50.0)123
5~2.6(2/77)1(50.0)1001(50.0)010
20~3.5(23/653)14(60.9)01139(39.1)360
40~4.2(18/433)10(55.6)3708(44.4)350
60~914.2(18/433)11(61.1)2637(38.9)241
合计73428268319184
注:a: χ2=49.4,P < 0.05; b: χ2=0.0,P>0.05; c: χ2=1.2,P>0.05。
2.2 气单胞菌腹泻病例的菌型特征

73例气单胞菌腹泻病例包括42例单一型感染和31例混合型感染病例,温和气单胞菌病例(分别为26例和18例)与其他菌型病例阳性率差异有统计学意义(P <0.05)。混合感染病例的多病原谱分布以2种病原菌的混合病例最多(93.5%),最多的为4种病原菌的混合病例;在混合感染的病原菌中以5种致泻性大肠埃希菌的频率最高(74.2%),其次为弯曲菌和沙门菌,见表 2

表2 气单胞菌腹泻病例的菌型分布特征 Table 2 Characteristics of Aeromonas infection
感染类型嗜水(n=17)温和(n=44)豚鼠(n=12)
单一型(n=42)826a8
混合型(n=31)918b4
混合感染的病原谱3(嗜水+EAEC)6(温和+ETEC)2(豚鼠+EAEC)
2(嗜水+ETEC)3(温和+EAEC)1(豚鼠+ETEC)
1(嗜水+EPEC)2(温和+EPEC)1(豚鼠+EPEC)
1(嗜水+沙门菌)2(温和+沙门菌)
1(嗜水+空肠弯曲菌)2(温和+空肠弯曲菌)
1(嗜水+结肠弯曲菌)1(温和+类志贺邻单胞菌)
1(温和+ETEC+EPEC)
1(温和+ETEC+EPEC+EAEC)
2.3 气单胞菌腹泻株的抗生素耐药特征

73株气单胞菌腹泻株中有48株(65.8%)至少对1种抗生素产生耐药性,对AMC和TE的耐药率最高,分别为38.4%和34.2%,其他10种抗生素的耐药率均 < 10.0%。出现2种以上抗生素耐药的菌型以温和气单胞菌为主(P <0.05),有3株温和气单胞菌对3种及以上抗生素产生了多重耐药(MDR),其中1株耐6种抗生素(AMC+TE+SXT+CTX+CIP+LEV),见表 3

表3 气单胞菌腹泻株的耐药特征 Table 3 ntibiotic resistance of Aeromonas strains
耐药型抗生素种类总耐药率(%,n=73)嗜水(n=17)温和(n=44)豚鼠(n=12)
R-1AMC23.3(17)854
TE19.2(14)1112
CTX2.7(2)011
SXT1.4(1)100
R-2TE+AMC8.2(6)132
AMC+IPM1.4(1)010
C+SXT1.4(1)010
CTX+TE1.4(1)100
SXT+AMC1.4(1)010
TE+SXT1.4(1)010
R-3TE+SXT+AMC2.7(2)020
R-6CTX+CIP+LEV+TE+SXT+AMC1.4(1)010
合计65.8(48)1227a9
注:a: χ2=12.5, P < 0.05。
3 讨论

随着分子测序技术在气单胞菌鉴定中的应用,近几年新发现的来自人源和环境源的气单胞菌新菌型数不断增多[5]。与腹泻相关的3种传统气单胞菌(型)病例在全球的分布仍存在较大的差异,这也是导致其未能获得普遍公认的肠道病原菌地位的原因之一。已公认的在儿童腹泻中占有相对重要地位的高发病地区,局限在南亚和东南亚的某些发展中国家[6, 7]。本次研究获得的上海浦东地区气单胞菌腹泻阳性率为4.0%,和国内北京(3.9%)、印度(4.7%)及西班牙(4.0%)的报道相近,说明其在多数国家和地区的发病基线总体较稳定[6, 8, 9]

本次研究尚存在局限性,即未使用病例对照方法求证气单胞菌腹泻的归因,但发现了气单胞菌腹泻也呈现典型的季节性和低年龄组与中老年人群易感的特征,符合通常的食源性感染性疾病的流行规律;虽然与其他肠道病原菌混合感染比例较高,也符合其不同菌型的致病性差异和人群免疫力强弱导致的症状以轻度或中度腹泻为主的临床特征,因此目前已有多国学者认为可将气单胞菌纳入食源性感染性病原菌范畴[7, 10]。国内沿海地区的研究已有证据认为气单胞菌和腹泻存在正相关[11]

本次的研究证实上海浦东地区的气单胞菌腹泻病例中以温和气单胞菌为优势的流行特征。不同于印度、西班牙等国的腹泻病例以豚鼠气单胞菌占优势(45.1%~75.9%)的特征[6, 9]。基于早期的气单胞菌食源性监测数据发现,浦东地区的食源性气单胞菌型分布也是以温和气单胞菌为优势,说明病例存在食源性暴露的潜在风险[12]。由此证明,美国环保总署(EPA)将气单胞菌列入水质污染指示菌名录,进一步提示其具有潜在的和食品相关的致病性。

细菌耐药性问题一直是全球关注的焦点,与人类健康息息相关。有研究显示,气单胞菌能产生头孢菌素酶、金属酶等[1]。自2003年法国首次报道产ESBLs的气单胞菌之后,世界各地陆续报道环境及临床分离的高耐药菌株,提示此类菌的耐药性问题日趋严重[13, 14, 15]。本次的人源气单胞菌耐药特征显示,65.8%的菌株至少对1种抗生素产生耐药性,与我国其他地区的报告一致[16]。耐药以青霉素酶抑制剂和四环素类药物最高,可能与水产品养殖业过程中的不合理使用抗生素添加剂促使菌株耐药性的升高,继而以垂直或水平方式在种群内传播的耐药机制有关[17]。本次确认1株对喹诺酮类(CIP)和三代头孢菌素类(CTX)等多重耐药的温和气单胞菌,其是否为ESBLs尚有待后续研究证实,但国外报道在养殖贻贝行业存在高产ESBLs菌株现象[18]。温和气单胞菌在浦东新区的人源和食源性气单胞菌中均属优势菌型,存在潜在的不断增加的多重耐药优势种群的风险,建议应加强食源性感染性疾病的风险评估,加强农业与食品安全流通领域的多源头协调监测,规范临床合理使用抗生素,降低多重耐药导致的疾病负担和暴发风险。

参考文献
[1] Janda JM, Abbott SL. The genus Aeromonas:taxonomy, pathogenicity, and infection[J]. Clin Microbiol Rev,2010,23(1):35-73.
[2] Sen K, Rodgers M. Distribution of six virulence factors in Aeromonas species isolated from US drinking water utilities:a PCR identification[J]. J Appl Microbiol,2004,97(5):1077-1086.
[3] Fu YF, Sun Q, Ye CC, et al. Etiology of acute diarrhea in Pudong New Area, Shanghai,2012[J]. Disease Surveillance,2014,29(5):349-353. (in Chinese)傅益飞,孙乔,叶楚楚, 等. 2012年上海市浦东新区急性腹泻门诊病例病原谱特征分析[J]. 疾病监测,2014,29(5):349-353.
[4] Clinical and Laboratory Standards Institute. Methods for antimicrobial dilution and disk susceptibility testing of infrequently isolated or fastidious bacteria; Approved guideline[S]. 2nd ed. Wayne, PA:Clinical and Laboratory Standards Institute,2010.
[5] Beaz-Hidalgo R, Latif-Eugenín F, Hossain MJ, 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.
[6] Sinha S, Shimada T, Ramamurthy T, et al. Prevalence, serotype distribution, antibiotic susceptibility and genetic profiles of mesophilic Aeromonas species isolated from hospitalized diarrhoeal cases in Kolkata, India[J]. J Med Microbiol,2004,53(6):527-534.
[7] Puthucheary SD, Puah SM, Chua KH. Molecular characterization of clinical isolates of Aeromonas species from Malaysia[J]. PLoS One,2012,7(2):e30205.
[8] Zhang Y, Zhang LR, Zhen BJ, et al. Molecular epidemiology characteristics of diarrhea Aeromonas hydrophila from Tongzhou, Beijing, in 2011[J]. Chinese Journal of Microecology,2013,25(12):1451-1453. (in Chinese)张扬,张兰荣,甄博珺,等. 2011年通州区腹泻源性嗜水气单胞菌的分子分型特征[J]. 中国微生态学杂志,2013,25(12):1451-1453.
[9] Pablos M, Remacha MA, Rodríguez-Calleja JM, et al. Identity, virulence genes, and clonal relatedness of Aeromonas isolates from patients with diarrhea and drinking water[J]. Eur J Clin Microbiol Infect Dis,2010,29(9):1163-1172.
[10] Senderovich Y, Ken-Dror S, Vainblat I, et al. A molecular study on the prevalence and virulence potential of Aeromonas spp. recovered from patients suffering from diarrhea in Israel[J]. PLoS One,2012,7(2):e30070.
[11] Chen X, Chen Y, Yang Q, et al. Plesiomonasshigelloides infection in Southeast China[J]. PLoS One,2013,8(11):e77877.
[12] Wang WQ, Su JH, Fu HQ, et al. Monitoring and phenotype characteristics study on food borne Aeromonas species[J]. Chinese Journal of Food Hygiene,2014,26(1):1-5. (in Chinese)王闻卿,苏靖华,傅慧琴,等. 食源性气单胞菌属种水平监测和表型特征研究[J]. 中国食品卫生杂志,2014,26(1):1-5.
[13] Marchandin H, Godreuil S, Darbas H, et al. Extended-spectrum β-lactamase TEM-24 in an Aeromonas clinical strain:acquisition from the prevalent Enterobacteraerogenes clone in France[J]. Antimicrob Agents Chemother,2003,47(12):3994-3995.
[14] Girlich D, Poirel L, Nordmann P. Diversity of clavulanic acid-inhibited extended-spectrumβ-lactamases in Aeromonas spp. from the Seine River, Paris, France[J]. Antimicrob Agents Chemother,2011,55(3):1256-1261.
[15] Wu CJ, Chuang YC, Lee MF, et al. Bacteremia due to extended-spectrum-β-lactamase-producing Aeromonas spp. at a Medical Center in Southern Taiwan[J]. Antimicrob Agents Chemother,2011,55(12):5813-5818.
[16] Li FJ, Wang WQ, Zhu ZQ, et al. Distribution, virulence-associated genes and antimicrobial resistance of Aeromonas isolates from diarrheal patients and water, China[J]. J Infect,2015,70(6):600-608.
[17] Yano Y, Hamano K, Tsutsui I, et al. Occurrence, molecular characterization, and antimicrobial susceptibility of Aeromonas spp. in marine species of shrimps cultured at inland low salinity ponds[J]. Food Microbiol,2015,47:21-27.
[18] Maravi A, Sko ibuši M, Šamani I, et al. Aeromonas spp. simultaneously harbouring blaCTX-M-15, blaSHV-12,blaPER-1 and blaFOX-2, in wild-growing Mediterranean mussel (Mytilusgalloprovincialis) from Adriatic Sea, Croatia[J]. Int J Food Microbiol,2013,166(2):301-308.