奶源异质性耐药大肠杆菌及其耐药亚群特性研究

doi:10.11843/j.issn.0366-6964.2024.12.043

Abstract

Abstract:

The aim of this study was to investigate whether heteroresistance exists in milk-derived Escherichia coli, and to characterize the biology of heteroresistance E. coli and its resistant subgroups, in order to explore possible heteroresistance mechanisms.The minimum inhibitory concentration (MIC) of one E. coli strain named D1 which isolated from raw cow's milk was determined by micro broth dilution method. The heteroresistance of E. coli was initially screened by the K-B paper diffusion method, and confirmed by the population analysis profile (PAP), and the heteroresistance characteristics of E. coli were investigated by resistance stability test and biofilm assay, finally, the mechanism of heterogeneous resistance was analyzed by whole genome sequencing and resequencing. The results showed that this E. coli was intermediary to polymyxin E, resistant to sulfisoxazole, and showed sensitivity to the rest of the antibiotics, and two E. coli-antimicrobial heteroresistance combinations were screened out by K-B paper diffusion assay.PAP confirmation showed that the MIC/maximum noninhibitory concentration (MNIC) ratio of E. coli was 8, and the occurence frequency of the resistant subgroups was 2.45×10^-6, confirming D1 to be a heteroresistance doxycycline-resistant strain. The stability of transmission showed that the resistant subpopulation could not be inherited stably, while there was no growth lag in the growth test, and the biofilm production of the resistant subpopulation was significantly greater than that of the parental strain. Whole genome sequencing revealed that there was a gene mutation between the resistant subpopulation and the parental strain, and the mutant gene txR is a specific transcriptional regulator that may lead to heteroresistance to doxycycline in E. coli. In conclusion, the existence of heteroresistance to doxycycline in E. coli from milk sources suggests that doxycycline antibiotics should be used rationally in the clinic, therefore, more attention should be paid to the occurrence of heteroresistance when using doxycycline in clinical treatment, and effective detection methods need to be used to provide guidance for clinical prevention and control and rational use of antibiotics.

Key words: milk-derived Escherichia coli, heteroresistance, resistant subgroups, whole genome

CLC Number:

S852.612

GAO Jiaojiao, ZHENG Nan, SHAO Wei, CHEN He, MA Xianlan, ZHAO Yankun. Characterization of Heterogeneous Drug-resistant Escherichia coli and Its Drug-resistant Subpopulations from Milk Sources[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(12): 5813-5824.

Figures/Tables 13

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References 35

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抗菌药物 Antimicrobial	MIC折点/(μg·mL^-1) MIC folding point			D1菌株MIC/(μg·mL^-1) D1 Antimicrobial Strain MIC	药敏类型 Type of drug sensitivity
抗菌药物 Antimicrobial	S	I	R	D1菌株MIC/(μg·mL^-1) D1 Antimicrobial Strain MIC	药敏类型 Type of drug sensitivity
氨苄西林 Ampicillin	≤8	16	≥32	1	S
阿莫西林/克拉维酸 Amoxicillin/clavulanic acid	≤8/4	16/8	≥32/16	2/1	S
头孢噻吩 Cephalothin	-	-	-	8	-
头孢噻呋 Ceftiofur	≤2	4	≥8	≤0.25	S
美罗培南 Meropenem	≤1	2	≥4	≤0.25	S
链霉素 Straptomycin	-	-	-	4	-
卡那霉素 Kanamycin	≤16	32	≥64	4	S
庆大霉素 Gentamicin	≤4	8	≥16	0.5	S
四环素 Tetracycline	≤4	8	≥16	≤0.25	S
多西环素 Doxycycline	≤4	8	≥16	0.25	S
氟苯尼考 Florfenicol	≤4	8	≥16	4	S
多粘菌素 E Colistin	-	≤2	≥4	0.5	I
利福昔明 Rifaximin	-	-	-	4	-
环丙沙星 Ciprofloxacin	≤0.25	0.5	≥1	≤0.25	S
磺胺异恶唑 Sulfafurazole	≤256	-	≥512	1 024	R
复方新诺明 Sulfamethoxazole	≤2/38	-	≥4/76	≤0.12/2.4	S

抗生素种类 Antibiotic type	耐药基因 Drug-resistance gene
氨基香豆素抗菌药物 Aminocoumarin antibiotic	gyrB、tolC、baeR、baeS、mdtC、mdtB、mdtA、cpxA
膦酸抗菌药物 Phosphonic acid antibiotic	uhpA、uhpT、murA、ptsI、glpT、mdtG、cyaA
氟喹诺酮类抗菌药物 Fluoroquinolone antibiotic	gadX、mdtF、mdtE、CRP、acrF、acrE、acrS、tolC、parE、parC、emrB、emrA、emrR、evgS、evgA、gyrA、mipA、H-NS、mdtH、acrA、acrB、mdtM、soxR、lamB
大环内酯类抗菌药物 Macrolide antibiotic	gadX、mdtF、mdtE、CRP、tolC、evgS、evgA
头孢菌素 Cephamycin	acrF、acrE、acrS、tolC、marA、H-NS、ompF、acrA、acrB、EC-15、soxR、soxS
噁唑烷酮类抗菌药物 Oxazolidinone antibiotic	mlaF、mlaD
肽抗菌药物 Peptide antibiotic	bacA、pmrF、yojI、ugd、eptA
氨基糖苷类抗菌药物 Aminoglycoside antibiotic	tolC、acrD、baeR、baeS、mipA、kdpE、cpxA
四环素抗菌药物 Tetracycline antibiotic	tolC、evgS、evgA、emrK、emrY、mdfA、lamB
碳青霉烯类抗菌药物 Carbapenem antibiotic	tolC、marA、ompF、soxS
核苷类抗菌药物 Nucleoside antibiotic	leuO、mdtN、mdtO、mdtP

分类 type	耐药基因 drug-resistance gene
耐药结节分化家族(RND) Resistant Nodal Differentiation Family (RND)	acrA、acrB、acrD、acrE、acrF、acrR、acrS、baeR、baeS、cpxA、CRP、evgA、evgS、gadX、H-NS、marA、marR、mdtA、mdtB、mdtC、mdtE、mdtF、soxR、soxS、tolC
主要易化子超家族(MFS) Major facilitator superfamily (MFS)	emrA、emrB、emrK、emrR、emrY、evgA、evgS、H-NS、mdfA、mdtG、mdtH、mdtM、mdtN、mdtO、mdtP、leuO、soxR、soxS、tolC
ATP结合盒家族超家族(ABC) ATP-binding cassette family superfamily (ABC)	msbA、mlaD、mlaF、soxR、soxS、tolC、yojl

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Characterization of Heterogeneous Drug-resistant Escherichia coli and Its Drug-resistant Subpopulations from Milk Sources

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