Analysis of biofilm genes and quorum sensing genes of carbapenem resistant <i>Acinetobacter baumannii</i> in the wounds of diabetic foot patients

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

Volume 40 Issue 12

Dec. 2024

Turn off MathJax

Article Contents

Article Navigation > CHINESE JOURNAL OF BURNS AND WOUNDS > 2024 > 40(12): 1166-1175

Xu J,Han XC,He L,et al.Analysis of biofilm genes and quorum sensing genes of carbapenem resistant Acinetobacter baumannii in the wounds of diabetic foot patients[J].Chin J Burns Wounds,2024,40(12):1166-1175.DOI: 10.3760/cma.j.cn501225-20240715-00269.

Citation:

Xu J,Han XC,He L,et al.Analysis of biofilm genes and quorum sensing genes of carbapenem resistant Acinetobacter baumannii in the wounds of diabetic foot patients[J].Chin J Burns Wounds,2024,40(12):1166-1175.DOI: 10.3760/cma.j.cn501225-20240715-00269.

Citation:

PDF( 2855 KB)

Analysis of biofilm genes and quorum sensing genes of carbapenem resistant Acinetobacter baumannii in the wounds of diabetic foot patients

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, Tianjin Medical University Chu Hsien-I Memorial HospitalInstitute of Endocrinology,Tianjin300134, China
2.
Department of Endocrinology/Rheumatology and Immunology, the Sixth Division Hospital, Xinjiang Production and Construction Corps, Wujiaqu831300, China
3.
Department of Infectious Medicine,the Sixth Division Hospital, Xinjiang Production and Construction Corps, Wujiaqu831300, China
4.
National Health Commission Key Laboratory of Hormones and Development, Tianjin Key Laboratory of Metabolic Diseases, Clinical Laboratory, Tianjin Medical University Chu Hsien-I Memorial HospitalInstitute of Endocrinology, Tianjin300134, China
5.
Institute of Antibiotics, Huashan Hospital, Fudan University, Shanghai200040, China

Funds:

General Program of National Natural Science Foundation of China 81973614

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

More Information

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

Abstract

Abstract

Objective To investigate the biofilm genes and quorum sensing genes of carbapenem resistant Acinetobacter baumannii (CRAB) in the wounds of diabetic foot patients. Methods This study was a retrospective observational study. The 233 strains of Acinetobacter baumannii were cultured from 177 inpatients (128 males and 49 females, aged (56±10) years) with diabetic foot admitted to the Department of Diabetic Foot of Tianjin Medical University Chu Hsien-I Memorial Hospital from October 2020 to September 2023. Two hundred and thirty-three Acinetobacter baumannii strains were detected by bacterial culture from the diabetic foot wounds of the aforementioned patients. All Acinetobacter baumannii strains were identified using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, followed by analysis of their resistance rate using kinetic turbidimetric assay by a fully automated microbiological analysis system. Ten CRAB strains (from 10 patients, 9 males and 1 female, aged (63±13) years) and 10 carbapenem sensitive Acinetobacter baumannii (CSAB; from 10 patients, 8 males and 2 females, aged (63±9) years) strains were randomly selected, and the whole DNA genome was extracted and sequenced. The antibiotic resistance genes were annotated using a comprehensive antibiotic resistance gene database, and a phylogenetic tree was drawn to analyze the homologous relationship between CRAB and CSAB. The 7 housekeeping genes of Acinetobacter baumannii was entered into the PubMLST website to analyze the multi-locus sequence typing of CRAB and CSAB. All the measured genes were put into the PubMLST website to search for the biofilm genes bap, csuA, csuB, csuA/B, csuC, csuD, csuE, pgaA, pgaB, pgaC, pgaD, bfmR, bfmS, ompA carried by each Acinetobacter baumannii, as well as the quorum sensing genes abaI and abaR, and flagellar gene pilA. The differences in carrying these genes between CRAB and CSAB were compared. The biofilm genes and quorum sensing genes carried by CRAB and CSAB carrying oxacillinase (OXA) resistance gene bla_OXA were analyzed. Gross observation was performed to check if there was gel-like membrane-like substance in the diabetic foot wounds infected with CRAB and CSAB, and if so, 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 pan-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 fully drug-resistant Acinetobacter baumannii was detected. Among 233 strains of Acinetobacter baumannii, the resistance rate to carbapenem antibiotics exceeded 80%; the resistance rate of cefoperazone/sulbactam was relatively low, at 37%; the resistance rates to the 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%; the resistance rates to tigecycline and colistin did not exceed 1%. The phylogenetic tree showed that 10 CRAB strains were highly homologous, while 10 CSAB strains had low homology. The analysis of multi-locus sequence typing showed that 10 CRAB strains were all the same type; among the 10 CSAB strains, except 1 strain without typing, the remaining 9 CSAB strains had 7 types. Eight of 10 CRAB strains contained complete biofilm genes and quorum sensing genes. The biofilm 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 (P<0.05). The main bla_OXA categories carried by CRAB were bla_OXA-23-like (specifically Bla_OXA-167) and bla_OXA-51-like (specifically bla_OXA-66), both of which had carbapenase activity. Eight of 10 CRAB strains carried both bla_OXA-66 and bla_OXA-167, and all of them had relatively complete quorum sensing genes and biofilm genes. The main bla_OXA categories carried by CSAB were bla_OXA-51-like and bla_OXA-213-like. Although they had carbapenemase activity, clinical drug sensitivity test showed that they were all sensitive to carbapenem antibiotics. 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 terms of multi-locus sequence typing, carrying biofilm genes, quorum sensing genes, and bla_OXA gene, leading to differences in antibiotic resistance between the two.
- Diabetic foot,
- Biofilms,
- Quorum sensing,
- Acinetobacter baumannii,
- Carbapenem,
- Drug resistance

FullText(HTML)

References(55)

References

[1]	吴静,郭立新.中国糖尿病地图[M].北京:人民卫生出版社,2022.
[2]	ChenL, SunS, GaoY, et al. Global mortality of diabetic foot ulcer: a systematic review and meta-analysis of observational studies[J]. Diabetes Obes Metab, 2023,25(1):36-45. DOI: 10.1111/dom.14840.
[3]	王宁, 鞠上. 糖尿病足溃疡难愈合机制研究进展[J].中华烧伤与创面修复杂志,2022,38(11):1085-1089. DOI: 10.3760/cma.j.cn501225-20220227-00038.
[4]	陈耀楠,王丹钰,袁倩,等. 糖尿病足溃疡慢性伤口的形成机制及新型敷料的研究进展[J]. 中华糖尿病杂志,2023,15(2):199-203. DOI: 10.3760/cma.j.cn115791-20220419-00173.
[5]	郭庆娇, 欧阳静, 饶佳琴, 等. 糖尿病患者糖尿病足溃疡复发风险预测模型的构建及初步验证[J].中华烧伤与创面修复杂志,2023,39(12):1149-1157. DOI: 10.3760/cma.j.cn501225-20231101-00166.
[6]	中华医学会糖尿病学分会. 中国2型糖尿病防治指南(2020年版)[J].中华糖尿病杂志,2021,13(4):315-409. DOI: 10.3760/cma.j.cn115791-20210221-00095.
[7]	BusSA, LaveryLA, Monteiro-SoaresM, et al. Guidelines on the prevention of foot ulcers in persons with diabetes (IWGDF 2019 update)[J]. Diabetes Metab Res Rev, 2020,36Suppl 1:e3269. DOI: 10.1002/dmrr.3269.
[8]	WangA, LvG, ChengX, et al. Guidelines on multidisciplinary approaches for the prevention and management of diabetic foot disease (2020 edition)[J/OL]. Burns Trauma, 2020,8:tkaa017[2024-07-15]. https://pubmed.ncbi.nlm.nih.gov/32685563/. DOI: 10.1093/burnst/tkaa017.
[9]	NelsonRE, HyunD, JezekA, et al. Mortality, length of stay, and healthcare costs associated with multidrug-resistant bacterial infections among elderly hospitalized patients in the United States[J]. Clin Infect Dis, 2022,74(6):1070-1080. DOI: 10.1093/cid/ciab696.
[10]	DuF, MaJ, GongH, et al. Microbial infection and antibiotic susceptibility of diabetic foot ulcer in China: literature review[J]. Front Endocrinol (Lausanne), 2022,13:881659. DOI: 10.3389/fendo.2022.881659.
[11]	中国两网监测云-全国细菌耐药监测网2024-07-15https://carss.cn/sys/Htmls/dist/index.html 中国两网监测云-全国细菌耐药监测网[EB/OL].[2024-07-15]. https://carss.cn/sys/Htmls/dist/index.html.
[12]	XuJ, ChenW, HeL, et al. Most postoperative reserved "normal" metatarsal stumps of diabetic foot osteomyelitis are infected but have healing potential[J]. Front Endocrinol (Lausanne), 2023,14:1165305. DOI: 10.3389/fendo.2023.1165305.
[13]	SennevilleE, GachetB, BlondiauxN, et al. Do anti-biofilm antibiotics have a place in the treatment of diabetic foot osteomyelitis?[J]. Antibiotics (Basel), 2023,12(2):317.DOI: 10.3390/antibiotics12020317.
[14]	MeaHJ, YongP, WongEH. An overview of Acinetobacter baumannii pathogenesis: motility, adherence and biofilm formation[J]. Microbiol Res, 2021,247:126722. DOI: 10.1016/j.micres.2021.126722.
[15]	YamabeK, ArakawaY, ShojiM, et al. Enhancement of Acinetobacter baumannii biofilm growth by cephem antibiotics via enrichment of protein and extracellular DNA in the biofilm matrices[J]. J Appl Microbiol, 2022,133(3):2002-2013. DOI: 10.1111/jam.15712.
[16]	TammaPD, AitkenSL, BonomoRA, et al. Infectious diseases society of america guidance on the treatment of ampC β-lactamase-producing enterobacterales, carbapenem-resistant Acinetobacter baumannii, and Stenotrophomonas maltophilia infections[J]. Clin Infect Dis, 2022,74(12):2089-2114. DOI: 10.1093/cid/ciab1013.
[17]	ChandranS, ManokaranY, VijayakumarS, et al. Enhanced bacterial killing with a combination of sulbactam/minocycline against dual carbapenemase-producing Acinetobacter baumannii[J]. Eur J Clin Microbiol Infect Dis,2023,42(5):645-651. DOI: 10.1007/s10096-023-04583-z.
[18]	HillyerT, BeninBM, SunC,et al. A novel strategy to characterize the pattern of β-lactam antibiotic-induced drug resistance in Acinetobacter baumannii[J]. Sci Rep,2023,13(1):9177. DOI: 10.1038/s41598-023-36475-9.
[19]	WatkinsRR, BonomoRA. Sulbactam-durlobactam: a step forward in treating carbapenem-resistant Acinetobacter baumannii (CRAB) infections[J]. Clin Infect Dis, 2023,76(Suppl 2):S163-S165. DOI: 10.1093/cid/ciad093.
[20]	LeeCR, LeeJH, ParkM, et al. Biology of Acinetobacter baumannii: pathogenesis, antibiotic resistance mechanisms, and prospective treatment options[J]. Front Cell Infect Microbiol, 2017,7:55. DOI: 10.3389/fcimb.2017.00055.
[21]	KarageorgopoulosDE, FalagasME. Current control and treatment of multidrug-resistant Acinetobacter baumannii infections[J]. Lancet Infect Dis, 2008,8(12):751-762. DOI: 10.1016/S1473-3099(08)70279-2.
[22]	MonemS, Furmanek-BlaszkB, ŁupkowskaA, et al. Mechanisms protecting Acinetobacter baumannii against multiple stresses triggered by the host immune response, antibiotics and outside-host environment[J]. Int J Mol Sci,2020,21(15):5498. DOI: 10.3390/ijms21155498.
[23]	IovlevaA, McElhenyCL, FowlerEL, et al. In vitro activity of sulbactam-durlobactam against colistin-resistant and/or cefiderocol-non-susceptible, carbapenem-resistant Acinetobacter baumannii collected in U.S. hospitals[J]. Antimicrob Agents Chemother, 2024,68(3):e0125823. DOI: 10.1128/aac.01258-23.
[24]	GiannellaM, VialeP. Treating carbapenem-resistant Acinetobacter baumannii infections[J]. Lancet Infect Dis, 2023,23(9):994-995. DOI: 10.1016/S1473-3099(23)00203-7.
[25]	MillerWR, AriasCA. ESKAPE pathogens: antimicrobial resistance, epidemiology, clinical impact and therapeutics[J]. Nat Rev Microbiol, 2024,22(10):598-616. DOI: 10.1038/s41579-024-01054-w.
[26]	IovlevaA, MustaphaMM, GriffithMP, et al. Carbapenem-resistant Acinetobacter baumannii in U.S. hospitals: diversification of circulating lineages and antimicrobial resistance[J]. mBio, 2022,13(2):e0275921. DOI: 10.1128/mbio.02759-21.
[27]	BulachD, CarterGP, LiL, et al. The whole-genome molecular epidemiology of sequential isolates of Acinetobacter baumannii colonizing the rectum of patients in an adult intensive care unit of a tertiary hospital[J]. Microbiol Spectr, 2023,11(6):e0219123. DOI: 10.1128/spectrum.02191-23.
[28]	HamidianM, NigroSJ. Emergence, molecular mechanisms and global spread of carbapenem-resistant Acinetobacter baumannii[J]. Microb Genom, 2019,5(10):e000306. DOI: 10.1099/mgen.0.000306.
[29]	GuD, WuY, ChenK, et al. Recovery and genetic characterization of clinically-relevant ST2 carbapenem-resistant Acinetobacter baumannii isolates from untreated hospital sewage in Zhejiang province, China[J]. Sci Total Environ, 2024,916:170058. DOI: 10.1016/j.scitotenv.2024.170058.
[30]	PakharukovaN, MalmiH, TuittilaM, et al. Archaic chaperone-usher pili self-secrete into superelastic zigzag springs[J]. Nature, 2022,609(7926):335-340. DOI: 10.1038/s41586-022-05095-0.
[31]	RomeroM, MayerC, HeebS, et al. Mushroom-shaped structures formed in Acinetobacter baumannii biofilms grown in a roller bioreactor are associated with quorum sensing-dependent Csu-pilus assembly[J]. Environ Microbiol, 2022,24(9):4329-4339. DOI: 10.1111/1462-2920.15985.
[32]	AhmadI, NadeemA, MushtaqF, et al. Csu pili dependent biofilm formation and virulence of Acinetobacter baumannii[J]. NPJ Biofilms Microbiomes, 2023,9(1):101. DOI: 10.1038/s41522-023-00465-6.
[33]	de BreijA, GaddyJ, van der MeerJ, et al. CsuA/BABCDE-dependent pili are not involved in the adherence of Acinetobacter baumannii ATCC19606(T) to human airway epithelial cells and their inflammatory response[J]. Res Microbiol, 2009,160(3):213-218. DOI: 10.1016/j.resmic.2009.01.002.
[34]	ChoiAH, SlamtiL, AvciFY, et al. The pgaABCD locus of Acinetobacter baumannii encodes the production of poly-beta-1-6-N-acetylglucosamine, which is critical for biofilm formation[J]. J Bacteriol, 2009,191(19):5953-5963. DOI: 10.1128/JB.00647-09.
[35]	WuHJ, XiaoZG, LvXJ, et al. Drug-resistant Acinetobacter baumannii: from molecular mechanisms to potential therapeutics (Review)[J]. Exp Ther Med, 2023,25(5):209. DOI: 10.3892/etm.2023.11908.
[36]	YangCH, SuPW, MoiSH,et al. Biofilm formation in Acinetobacter baumannii: genotype-phenotype correlation[J]. Molecules, 2019,24(10):1849. DOI: 10.3390/molecules24101849.
[37]	HardingCM, HennonSW, FeldmanMF. Uncovering the mechanisms of Acinetobacter baumannii virulence[J]. Nat Rev Microbiol, 2018,16(2):91-102. DOI: 10.1038/nrmicro.2017.148.
[38]	MendesSG, ComboSI, AllainT, et al. Co-regulation of biofilm formation and antimicrobial resistance in Acinetobacter baumannii: from mechanisms to therapeutic strategies[J]. Eur J Clin Microbiol Infect Dis, 2023,42(12):1405-1423. DOI: 10.1007/s10096-023-04677-8.
[39]	KimSY, KimMH, KimSI, et al. The sensor kinase BfmS controls production of outer membrane vesicles in Acinetobacter baumannii[J]. BMC Microbiol, 2019,19(1):301. DOI: 10.1186/s12866-019-1679-0.
[40]	TiwariV, PatelV, TiwariM. In-silico screening and experimental validation reveal L-Adrenaline as anti-biofilm molecule against biofilm-associated protein (Bap) producing Acinetobacter baumannii[J]. Int J Biol Macromol, 2018,107(Pt A):1242-1252. DOI: 10.1016/j.ijbiomac.2017.09.105.
[41]	UpmanyuK, KumarR, Rizwanul HaqueQM, et al. Exploring the evolutionary and pathogenic role of Acinetobacter baumannii biofilm-associated protein (Bap) through in silico structural modeling[J]. Arch Microbiol,2024,206(6):267.DOI: 10.1007/s00203-024-03992-8.
[42]	MohamadTS, RahmanJK, AhmedAA, et al. Down-regulation of abaI, abaR, Bap and OmpA genes in Acinetobacter baumannii by ethanol extract of Glycyrrhiza glabra after toxicity assessment[J]. Cell Mol Biol (Noisy-le-grand),2023,69(12):194-200. DOI: 10.14715/cmb/2023.69.12.31.
[43]	LiouML, SooPC, LingSR, et al. The sensor kinase BfmS mediates virulence in Acinetobacter baumannii[J]. J Microbiol Immunol Infect, 2014,47(4):275-281. DOI: 10.1016/j.jmii.2012.12.004.
[44]	KimHJ, KimNY, KoSY, et al. Complementary regulation of BfmRS two-component and AbaIR quorum sensing systems to express virulence-associated genes in Acinetobacter baumannii[J]. Int J Mol Sci, 2022,23(21):13136. DOI: 10.3390/ijms232113136.
[45]	LiY, WangB, LuF, et al. Synergistic inhibitory effect of polymyxin B in combination with ceftazidime against robust biofilm formed by Acinetobacter baumannii with genetic deficiency in AbaI/AbaR quorum sensing[J]. Microbiol Spectr, 2022,10(1):e0176821. DOI: 10.1128/spectrum.01768-21.
[46]	PaluchE, Rewak-SoroczyńskaJ, JędrusikI, et al. Prevention of biofilm formation by quorum quenching[J]. Appl Microbiol Biotechnol, 2020,104(5):1871-1881. DOI: 10.1007/s00253-020-10349-w.
[47]	LawSKK, TanHS. The role of quorum sensing, biofilm formation, and iron acquisition as key virulence mechanisms in Acinetobacter baumannii and the corresponding anti-virulence strategies[J]. Microbiol Res, 2022,260:127032. DOI: 10.1016/j.micres.2022.127032.
[48]	LiH, LiuF, ZhangY, et al. Evolution of carbapenem-resistant Acinetobacter baumannii revealed through whole-genome sequencing and comparative genomic analysis[J]. Antimicrob Agents Chemother, 2015,59(2):1168-1176. DOI: 10.1128/AAC.04609-14.
[49]	HackelMA, TsujiM, YamanoY, et al. In vitro activity of the siderophore cephalosporin, cefiderocol, against a recent collection of clinically relevant Gram-negative bacilli from North America and Europe, including carbapenem-nonsusceptible isolates (SIDERO-WT-2014 study)[J]. Antimicrob Agents Chemother, 2017,61(9):e00093-17. DOI: 10.1128/AAC.00093-17.
[50]	LeeYL, KoWC, HsuehPR. Geographic patterns of Acinetobacter baumannii and carbapenem resistance in the Asia-Pacific Region: results from the Antimicrobial Testing Leadership and Surveillance (ATLAS) program, 2012-2019[J]. Int J Infect Dis, 2023,127:48-55. DOI: 10.1016/j.ijid.2022.12.010.
[51]	NigroSJ, HallRM. Does the intrinsic oxaAb (blaOXA-51-like) gene of Acinetobacter baumannii confer resistance to carbapenems when activated by ISAba1?[J]. J Antimicrob Chemother, 2018,73(12):3518-3520. DOI: 10.1093/jac/dky334.
[52]	AnggrainiD, KemalRA, HadiU, et al. The susceptibility pattern and distribution of blaOXA-23 genes of clinical isolate Acinetobacter baumannii in a tertiary hospital, Indonesia[J]. J Infect Dev Ctries,2022,16(5):821-826. DOI: 10.3855/jidc.15902.
[53]	AbouelhassanY, NicolauDP, AbdelraoufK. Defining optimal sulbactam regimens for treatment of Acinetobacter baumannii pneumonia and impact of blaOXA-23 on efficacy[J]. J Antimicrob Chemother, 2024,79(9):2306-2316. DOI: 10.1093/jac/dkae229.
[54]	NeupaneL, SahAK, RayamajheeB, et al. Detection of blaoxa-23 gene from carbapenem-resistant Acinetobacter baumannii[J]. J Nepal Health Res Counc,2023,20(4):899-905. DOI: 10.33314/jnhrc.v20i4.4257.
[55]	LiuS, HuangG, GongY, et al. Rapid and accurate detection of carbapenem-resistance gene by isothermal amplification in Acinetobacter baumannii[J/OL]. Burns Trauma,2020,8:tkaa026[2024-07-15]. https://pubmed.ncbi.nlm.nih.gov/32905076/. DOI: 10.1093/burnst/tkaa026.