Biofilm and quorum sensing genes analysis of carbapenem resistant Acinetobacter baumannii in diabetic foot wound

Xu Jun; Han Xiaocui; He Lu; Feng Shuhong; Sun Dongjian; Cao Chen; Liu Xijiao; Zhang Yanyan; Ding Baixing; Chang Bai

doi:10.3760/cma.j.cn501225-20240715-00269

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Article Navigation > CHINESE JOURNAL OF BURNS AND WOUNDS > > Accepted Manuscript

Xu Jun,Han Xiaocui,He Lu,et al.Biofilm and quorum sensing genes analysis of carbapenem resistant Acinetobacter baumannii in diabetic foot wound[J].Chin J Burns Wounds,2024,40(12):1-9.DOI: 10.3760/cma.j.cn501225-20240715-00269.

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Biofilm and quorum sensing genes analysis of carbapenem resistant Acinetobacter baumannii in diabetic foot wound

doi: 10.3760/cma.j.cn501225-20240715-00269

1.
National Health Commission Key Laboratory of Hormones and Development, Tianjin Key Laboratory of Metabolic Diseases, Department of Diabetic Foot, Chu HsienI Memorial HospitalTianjin Institute of Endocrinology, Tianjin Medical University, Tianjin300134, China
2.
Department of Endocrinology, the Sixth Division Hospital, Xinjiang Production and Construction Corps, Wujiaqu831300, China
3.
National Health Commission Key Laboratory of Hormones and Development, Tianjin Key Laboratory of Metabolic Diseases, Clinical Laboratory, Chu HsienI Memorial HospitalTianjin Institute of Endocrinology, Tianjin Medical University, Tianjin300134, China
4.
Institute of Antibiotics, Huashan Hospital, Fudan University, Shanghai200040, China

Funds:

General Program of National Natural Science Foundation of China 819724

Tianjin Key Medical Discipline (Speciaty) Construction Project TJYXZDXK-032A

More Information

Corresponding author: Chang Bai, Email: changbai1972@126.com
Received Date: 2024-07-15
Available Online: 2024-12-02

Abstract

Abstract

Objective To investigate the biofilm genes and quorum sensing genes of carbapenem resistant Acinetobacter baumannii (CRAB) in the wounds of diabetic foot and their effects on antibiotic resistance. Methods This study was an experimental study. The 233 strains of Acinetobacter baumannii cultured from 177 inpatients (128 males and 49 females, aged (56±10) years) with diabetic foot admitted to the Department of Diabetic Foot of Chu HsienI Memorial Hospital & Tianjin Institute of Endocrinology of Tianjin Medical University from October 2020 to September 2023. All Acinetobacter baumannii were subjected to antibiotic sensitivity testing using broth microdilution method, followed by identification and analysis of their resistance rate using a fully automated antibiotic sensitivity analyzer. Randomly select 10 strains of CRAB (from 10 patients, 9 males and 1 femles, aged (63±13) years) and 10 strains of carbapenem sensitive Acinetobacter baumannii (CSAB; from 10 patients, 8 males and 2 femles, aged (63±9) years) for whole genome sequencing. The antibiotic resistance genes were annotated using a comprehensive antibiotic resistance gene database, and a phylogenetic tree was draw to analyze the homologous relationship between CRAB and CSAB. The 7 housekeeping genes of Acinetobacter baumannii was entered into the website https://pubmlst.org to analyze the multilocus sequence typing between CRAB and CSAB. All the measured genes were putted into the PubMLST website to search for the biofilm genes bap, csuA, csuB, csuA/B, csuC, csuD, csuE, pagA, pagB, pagC, pagD, bfmR, bfmS, ompA carried by each Acinetobacter baumannii, as well as the quorum sensing genes abaI, abaR, and flagellar gene pilA. The differences in carrying these genes between CRAB and CSAB were compared. The correlation between biofilm genes, quorum sensing genes and Oxacillinase (OXA) resistance gene blaOXA from CRAB and CSAB was analyzed. Whether there was gel-like membrane-like substance in the wound of diabetic foot infected with CRAB and CSAB was observed by gross observation, and if there was, the microstructure was observed by scanning electron microscope. Results Among the detected Acinetobacter baumannii, the positive detection rates of CSAB, CRAB, multi-drug resistant Acinetobacter baumannii, and extensively-drug resistant Acinetobacter baumannii were 16.7% (39/233), 83.3% (194/233), 95.3% (222/233), and 34.3% (80/233), respectively, and no pan-drug resistant Acinetobacter baumannii was detected. Among 233 strains of Acinetobacter baumannii, the resistance rate of carbapenem antibiotics exceeded 80%; the resistance rates of cefoperazone/sulbactam was relatively low, at 37%; the resistance rates to other cephalosporin antibiotics (cefotaxime, ceftazimide, cefotetan, and cefuroxime) were more than 80%; the resistance rates to all penicillin antibiotics were greater than 80%; the resistance rates to quinolone antibiotics were all over 60%; the resistance rate to minocycline was only 12.0%; the resistance rates to tigecycline and colistin did not exceed 1%. The phylogenetic tree showed that 10 strains of CRAB were highly homologous, while 10 strains of CSAB had low homology. The analysis of multilocus sequence typing showed that 10 strains of CRAB strains were all the same type; except 1 strain without typing, the remaining 9 strains in 10 strains of CSAB had 7 types. Eight strains in 10 strains of CRAB contained complete biofilm genes and quorum sensing genes. The biofilm genes and quorum sensing genes from the strains of CSAB were incomplete and none carried the bap gene. Neither CRAB nor CSAB carried the flagellar gene pilA. Compared with that carried by CRAB, biofilm genes bap, csuA, csuC, and csuD and quorum sensing genes abaI and abaR carried by CSAB were significantly decreased (all P values <0.05). The main blaOXA categories carried by CRAB were BlaOXA-23-like (specifically BlaOXA-167) and BlaOXA-51-like (specifically BlaOXA-66), both of which had carbapenase activity. Eight of ten CRAB strains carried both blaOXA-66 and bla-OXA167, and all of them had relatively complete quorum sensing genes and biofilm genes. The main blaOXA categories carried by CSAB were BlaOXA-51-like and BlaOXA-213-like. Although they had carbapenemase activity, they were all sensitive to carbapenems in clinical drug sensitivity test. The biofilm genes carried by 10 strains of CSAB were incomplete, and only 3 strains of CSAB had complete quorum sensing genes. Gel-like and membrane-like substance could be seen in wounds infected with CRAB, which were biofilm; No gel-like and membrane-like substance was found in the wound infected with CSAB. Conclusions CRAB and CSAB in diabetic foot wounds are significantly different in multilocus sequence typing, carrying biofilm genes and quorum sensing genes, and the correlation with blaOXA gene, leading to differences in antibiotic resistance between the two.
- Diabetic foot,
- Biofilms,
- Quorum sensing,
- Acinetobacter baumannii,
- Carbapenem,
- Durg resistance

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References(35)

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