Antimicrobial resistance and resistance genes of Escherichia coli from pets
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摘要:目的
了解广州市宠物源大肠埃希菌Escherichia coli耐药性和耐药基因携带情况。
方法2016年7月至2017年7月从广州市4家宠物医院采集健康或患病犬猫样品共319份,其中,健康动物127份,患病动物192份。采用选择性培养基分离大肠埃希菌,利用基质辅助激光解析串联飞行时间质谱仪(MALDI-TOF MS)鉴定菌种;采用琼脂稀释法测定大肠埃希菌对11种抗菌药物的敏感性,利用PCR和测序检测耐药基因的携带情况。
结果319份样品共分离得到大肠埃希菌203株,其中,患病动物源109株,健康动物源94株。203株大肠埃希菌中有179株至少对1种抗生素耐药;对氨苄西林耐药率最高(76.85%),对头孢噻肟、四环素、多西环素和磺胺甲噁唑−甲氧苄啶耐药率均高于50%;对阿米卡星最为敏感,耐药率仅为10.84%。患病动物源大肠埃希菌对11种抗菌药物的耐药率均高于健康动物源,除阿米卡星、氟苯尼考和磷霉素外,对其他药物的耐药性均差异极显著(P < 0. 01)。耐药基因检测结果显示, floR 检出率最高(检出率为34.97%),blaCTX-M-9G、blaCTX-M-1G、fosA3、rmtB和blaCMY-2检出率分别为22.66%、20.19%、17.73%、10.34%和1.48%,未检测到blaCTX-M-2G和blaCTX-M-25G。
结论广州地区宠物源大肠埃希菌耐药状况严峻,且常携带多种重要耐药基因。应当加强对宠物源细菌耐药性的监测。
Abstract:ObjectiveTo investigate antimicrobial resistance and resistance genes of Escherichia coli from pet animals in Guangzhou.
MethodFrom July 2016 to July 2017, 319 samples were collected from cats and dogs at four animal hospitals in Guangzhou, including 127 samples from healthy animals and 192 samples from diseased animals. E. coli strains were isolated using the selective media, and were identified by MALDI-TOF mass spectrometry. The susceptibilities of E. coli isolates to 11 antimicrobial agents were determined by the agar dilution method. The presence of resistance genes was determined by PCR and sequencing.
ResultA total of 203 E. coli strains were isolated from 319 samples, including 109 E. coli isolates from diseased animals and 94 E. coli isolates from healthy animals. Among the 203 strains, 179 strains were resistant to at least one antimicrobial agent. The isolates showed the highest resistance rate against ampicillin (76.85%). The resistance rates against cefotaxime, tetracycline, doxycycline and sulfamethoxazole-trimethoprim were above 50%. The isolates were the most susceptible to amikacin with the resistance rate of only 10.84%. The isolates from diseased animals showed higher resistance rates against all antimicrobial agents compared with those from healthy animals. The resistance rates against all antimicrobial agents except amikacin, florfenicol and fosfomycin were significantly higher in the isolates from diseased animals compared with healthy animals (P<0.01). The detection results of resistance genes showed thatfloR had the highest detection rate (34.97%), the detection rates of blaCTX-M-9G, blaCTX-M-1G, fosA3, rmtB and blaCMY-2 genes were 22.66%, 20.19%, 17.73%,10.34% and 1.48% respectively. The genes blaCTX-M-2G and blaCTX-M-25G were not detected.
ConclusionThe antimicrobial resistance of E. coli isolates from pet animals in Guangzhou has become a serious problem, and some isolates carry several important resistance genes. Antimicrobial resistance in pet animals requires strict monitoring.
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Keywords:
- pet /
- Escherichia coli /
- antimicrobial resistance /
- resistance gene /
- antibiotic
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表 1 检测耐药基因的引物序列
Table 1 Sequences of primers used to screen resistance genes
基因
Gene引物序列(5′→3′)
Primer sequence片段大小/bp
Segment size参考文献
ReferenceblaCTX-M-1G F:CTTCCAGAATAAGGAATCCC 949 [7] R:CGTCTAAGGCGATAAACAAA blaCTX-M-2G F:ATGATGACTCAGAGCATTCG 857 [8] R:TCAGAAACCGTGGGTTACGA blaCTX-M-9G F:TGACCGTATTGGGAGTTTG 902 [7] R:ATGATGACTCAGAGCATTCG blaCTX-M-25G F:AATGTTCAGGGGATTAGGA 876 [7] R:ATCACTCCACATGGTGAGTA blaCMY-2 F:TGGCCGTTGCCGTTATCTAC 870 [9] R:CCCGTTTTATGCACCCATGA fosA3 F:GCGTCAAGCCTGGCATTT 282 [10] R:GCCGTCAGGGTCGAGAAA floR F:CTGAGGGTGTCGTCATCTAC 673 [9] R:GTCCCGACAATGCTGACTAT rmtB F:ATATCAACGATGCCCTCAC 725 [11] R:AAGTTCTGTTCCGATGGTC 表 2 203株大肠埃希菌耐药率
Table 2 Antimicrobial resistance rates of 203 Escherichia coli isolates
% 药物
Drug健康动物源(n=94)
Strains from healthy animals患病动物源1)(n=109)
Strains from sick animals全部菌株(n=203)
Total strains氨苄西林 AMP 65.96 86.24** 76.85 头孢噻肟 CTX 42.55 67.89** 56.16 头孢他啶 CAZ 12.77 31.19** 22.66 庆大霉素 GEN 28.72 46.79** 38.42 阿米卡星 AMI 7.45 13.76 10.84 四环素 TET 48.94 82.57** 67.00 多西环素 DOX 36.17 66.06** 52.22 氟苯尼考 FFC 35.11 44.95 40.39 环丙沙星 CIP 29.79 59.63** 45.81 磷霉素 FOS 32.98 34.86 33.99 磺胺甲噁唑−甲氧苄啶 SMZ-TMP 38.30 74.31** 57.64 1) “**”表示与健康动物源大肠埃希菌的耐药性差异极显著(P<0.01,卡方检验)
1) “**” indicates significant difference from drug resistance rate of Escherichia coli of healthy animals (P < 0.01, chi-square test)表 3 大肠埃希菌多重耐药情况
Table 3 Multi-drug resistance of Escherichia coli
耐药数
Number of drug resistance健康动物源(n=94)
Strains from healthy animals患病动物源(n=109)
Strains from sick animals所有菌株(n=203)
Total strains菌株数
Number of strains耐药率/%
Drug resisitance rate菌株数
Number of strains耐药率/%
Drug resisitance rate菌株数
Number of strains耐药率/%
Drug resisitance rate0 19 20.21 5 4.59 24 11.82 1 12 12.77 1 0.92 13 6.40 2 11 11.70 5 4.59 16 7.88 3 8 8.51 17 15.60 25 12.32 4 10 10.64 5 4.59 15 7.39 5 5 5.32 7 6.42 12 5.91 6 5 5.32 20 18.35 25 12.32 7 7 7.45 9 8.26 16 7.88 8 4 4.26 19 17.43 23 11.33 9 9 9.57 10 9.17 19 9.36 10 3 3.19 7 6.42 10 4.93 11 1 1.06 4 3.67 5 2.46 表 4 203株大肠埃希菌耐药基因检出率
Table 4 Detection rates of resistance genes from 203 Escherichia coli isolates
耐药基因
Resistance gene健康动物源(n=94)
Strains from healthy animals患病动物源(n=109)
Strains from sick animals所有菌株(n=203)
Total strains菌株数
Number of strains检出率/%
Detection rate菌株数
Number of strains检出率/%
Detection rate菌株数
Number of strains检出率/%
Detection rateblaCTX-M-1G 11 11.70 30 27.52 41 20.19 blaCTX-M-9G 14 14.89 32 29.36 46 22.66 blaCTX-M-2G 0 0 0 0 0 0 blaCTX-M-25G 0 0 0 0 0 0 floR 26 27.66 45 41.28 71 34.97 fosA3 14 14.89 22 20.28 36 17.73 rmtB 6 6.38 15 13.76 21 10.34 blaCMY-2 2 2.13 1 0.92 3 1.48 -
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