The increasing number of carbapenem-resistant Acinetobacter baumannii clinical isolates is a major concern, which restricts therapeutic options for treatment of serious infections caused by this emerging pathogen. The aim of this work is to assess the antimicrobial resistance profile and identify the molecular mechanisms involved in carbapenem resistance in A. baumannii isolated from different clinical sources in Mansoura University Hospitals, Egypt. Antimicrobial susceptibility testing has shown that resistance to carbapenem has dramatically increased (98%) with concomitant elevated levels of resistance to quinolones, trimethoprim/sulfamethoxazole, and aminoglycosides. Polymyxin B and colistin are considered the last resort. Random amplified polymorphic DNA (RAPD) typing method revealed great diversity among A. baumannii isolates. Coexistence of diverse intrinsic and acquired carbapenem-hydrolyzing β-lactamases has been detected in the tested isolates: Ambler class A: blaKPC (56%) and blaGES (48%), and Ambler class B: blaNDM (30%), blaSIM (28%), blaVIM (20%), and blaIMP (10%). Most isolates (94%) carried blaOXA-23-like and blaOXA-51-like simultaneously. blaOXA-23-like was preceded by ISAba1 providing a potent promoter activity for its expression. Sequencing analysis revealed that ISAba1 has been also inserted in carbapenem resistance-associated outer membrane protein (OMP) (carO) gene in three isolates, two of which were clonal based on RAPD typing, leading to interruption of its expression as confirmed by SDS-PAGE analysis of OMP fractions. Carbapenem resistance genes are widely distributed among A. baumannii clinical isolates from different clinical sources. Therefore, enhanced infection control measures, effective barriers, and rational use of antimicrobials should be enforced in hospitals for minimizing the widespread resistance to carbapenems and all other antibiotics.
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References
1.
PelegA.Y., SeifertH., and PatersonD.L.. 2008. Acinetobacter baumannii: emergence of a successful pathogen. Clin. Microbiol. Rev. 21:538–582.
2.
BonninR.A., NordmannP., and PoirelL.. 2013. Screening and deciphering antibiotic resistance in Acinetobacter baumannii: a state of the art. Expert Rev. Anti Infect. Ther. 11:571–583.
3.
PerezF., HujerA.M., HujerK.M., DeckerB.K., RatherP.N., and BonomoR.A.. 2007. Global challenge of multidrug-resistant Acinetobacter baumannii. Antimicrob. Agents Chemother. 51:3471–3484.
4.
KimY.A., ParkY.S., YoukT., LeeH., and LeeK.. 2018. Abrupt increase in rate of imipenem resistance in Acinetobacter baumannii complex strains isolated from general hospitals in Korea and correlation with carbapenem administration during 2002–2013. Ann. Lab. Med. 38:179–181.
5.
DafopoulouK., TsakrisA., and PournarasS.. 2018. Changes in antimicrobial resistance of clinical isolates of Acinetobacter baumannii group isolated in Greece, 2010–2015. J. Med. Microbiol. 67:496–498.
6.
EvansB.A., and AmyesS.G.. 2014. OXA beta-lactamases. Clin. Microbiol. Rev. 27:241–263.
7.
VilaJ., MartiS., and Sanchez-CespedesJ.. 2007. Porins, efflux pumps and multidrug resistance in Acinetobacter baumannii. J. Antimicrob. Chemother. 59:1210–1215.
8.
PoirelL., and NordmannP.. 2006. Carbapenem resistance in Acinetobacter baumannii: mechanisms and epidemiology. Clin. Microbiol. Infect. 12:826–836.
9.
TurtonJ.F., WardM.E., WoodfordN., KaufmannM.E., PikeR., LivermoreD.M., and PittT.L.. 2006. The role of ISAba1 in expression of OXA carbapenemase genes in Acinetobacter baumannii. FEMS Microbiol. Lett. 258:72–77.
10.
ColleJ.G., MilesR.S., and WattB.. 1996. Tests for identification of bacteria. In ColleJ.G., FraserA.G., MarimonB.P., SimmonA., Mackie and MacCartney (ed.), Practical medical microbiology. 14th ed. Churchill Livingstone, Edinburgh, pp. 151–179.
11.
BartualS.G., SeifertH., HipplerC., LuzonM.A., WisplinghoffH., and Rodriguez-ValeraF.. 2005. Development of a multilocus sequence typing scheme for characterization of clinical isolates of Acinetobacter baumannii. J. Clin. Microbiol. 43:4382–4390.
12.
CortivoG.D., GutberletA., FerreiraJ.A., FerreiraL.E., DeglmannR.C., WestphalG.A., and FrançaP.H.C.D.. 2015. Antimicrobial resistance profiles and oxacillinase genes in carbapenem-resistant Acinetobacter baumannii isolated from hospitalized patients in Santa Catarina, Brazil. Rev. Soc. Bras. Med. Trop. 48:699–705.
13.
TurtonJ.F., WoodfordN., GloverJ., YardeS., KaufmannM.E., and PittT.L.. 2006. Identification of Acinetobacter baumannii by detection of the blaOXA-51-like carbapenemase gene intrinsic to this species. J. Clin. Microbiol. 44:2974–2976.
14.
WangX., ChenT., YuR., LüX., and ZongZ.. 2013. Acinetobacter pittii and Acinetobacter nosocomialis among clinical isolates of the Acinetobacter calcoaceticus-baumannii complex in Sichuan, China. Diagn. Microbiol. Infect. Dis. 76:392–395.
15.
WenH., WangK., LiuY., TayM., LauroF.M., HuangH., WuH., LiangH., DingY., GivskovM., ChenY., YangL.. 2014. Population dynamics of an Acinetobacter baumannii clonal complex during colonization of patients. J. Clin. Microbiol. 52:3200–3208.
16.
HerasJ., DominguezC., MataE., PascualV., LozanoC., TorresC., and ZarazagaM.. 2015. GelJ—a tool for analyzing DNA fingerprint gel images. BMC Bioinformatics. 16:270.
17.
BauerA.W., KirbyW.M., SherrisJ.C., and TurckM.. 1966. Antibiotic susceptibility testing by a standardized single disk method. Am. J. Clin. Pathol. 45:493–496.
18.
Clinical and Laboratory Standards Institute (CLSI). Performance Standards for Antimicrobial Susceptibility Testing. 27th ed. CLSI document M100-S27 (ISBN 1-56238-805-3).
19.
YongD., LeeK., YumJ.H., ShinH.B., RossoliniG.M., and ChongY.. 2002. Imipenem-EDTA disk method for differentiation of metallo-beta-lactamase-producing clinical isolates of Pseudomonas spp. and Acinetobacter spp. J. Clin. Microbiol. 40:3798–3801.
20.
KhalilM.A.F., ElgamlA., and El-MowafyM.. 2017. Emergence of multidrug-resistant New Delhi metallo-beta-lactamase-1-producing Klebsiella pneumoniae in Egypt. Microb. Drug Resist. 23:480–487.
21.
TsakrisA., IkonomidisA., PournarasS., TzouvelekisL.S., SofianouD., LegakisN.J., and ManiatisA.N.. 2006. VIM-1 metallo-beta-lactamase in Acinetobacter baumannii. Emerg. Infect. Dis. 12:981–983.
22.
PoirelL., RevathiG., BernabeuS., and NordmannP.. 2011. Detection of NDM-1-producing Klebsiella pneumoniae in Kenya. Antimicrob. Agents Chemother. 55:934–936.
23.
MendesR.E., KiyotaK.A., MonteiroJ., CastanheiraM., AndradeS.S., GalesA.C., PignatariA.C., and TufikS.. 2007. Rapid detection and identification of metallo-beta-lactamase-encoding genes by multiplex real-time PCR assay and melt curve analysis. J. Clin. Microbiol. 45:544–547.
24.
DallenneC., Da CostaA., DecreD., FavierC., and ArletG.. 2010. Development of a set of multiplex PCR assays for the detection of genes encoding important beta-lactamases in Enterobacteriaceae. J. Antimicrob. Chemother. 65:490–495.
25.
El-ShazlyS., DashtiA., ValiL., BolarisM., and IbrahimA.S.. 2015. Molecular epidemiology and characterization of multiple drug-resistant (MDR) clinical isolates of Acinetobacter baumannii. Int. J. Infect. Dis. 41:42–49.
26.
WoodfordN., EllingtonM.J., CoelhoJ.M., TurtonJ.F., WardM.E., BrownS., AmyesS.G., and LivermoreD.M.. 2006. Multiplex PCR for genes encoding prevalent OXA carbapenemases in Acinetobacter spp. Int. J. Antimicrob. Agents. 27:351–353.
27.
HigginsP.G., LehmannM., and SeifertH.. 2010. Inclusion of OXA-143 primers in a multiplex polymerase chain reaction (PCR) for genes encoding prevalent OXA carbapenemases in Acinetobacter spp. Int. J. Antimicrob. Agents. 35:305.
28.
SegalH., GarnyS., and ElishaB.G.. 2005. Is IS(ABA-1) customized for Acinetobacter? FEMS Microbiol. Lett. 243:425–429.
29.
LimanskyA.S., MussiM.A., and VialeA.M.. 2002. Loss of a 29-kilodalton outer membrane protein in Acinetobacter baumannii is associated with imipenem resistance. J. Clin. Microbiol. 40:4776–4778.
30.
LaemmliU.K.1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 227:680–685.
31.
ZekaA.N., PoirelL., SipahiO.R., BonninR.A., ArdaB., OzinelM., UlusoyS., BorC., and NordmannP.. 2014. GES-type and OXA-23 carbapenemase-producing Acinetobacter baumannii in Turkey. J. Antimicrob. Chemother. 69:1145–1146.
32.
CorreaA., Del CampoR., Escandon-VargasK., PerenguezM., Rodriguez-BanosM., Hernandez-GomezC., PallaresC., PerezF., AriasC.A., CantonR., and VillegasM.V.. 2018. Distinct genetic diversity of carbapenem-resistant Acinetobacter baumannii from Colombian Hospitals. Microb. Drug Resist. 24:48–54.
33.
KarampatakisT., AntachopoulosC., TsakrisA., and RoilidesE.. 2017. Molecular epidemiology of carbapenem-resistant Acinetobacter baumannii in Greece: an extended review (2000–2015). Future Microbiol. 12:801–815.
34.
ChangY., LuanG., XuY., WangY., ShenM., ZhangC., ZhengW., HuangJ., YangJ., and JiaX.. 2015. Characterization of carbapenem-resistant Acinetobacter baumannii isolates in a Chinese teaching hospital. Front. Microbiol. 6:910.
35.
ChenY., GaoJ., ZhangH., and YingC.. 2017. Spread of the blaOXA-23-containing Tn2008 in carbapenem-resistant Acinetobacter baumannii isolates grouped in CC92 from China. Front. Microbiol. 8:163.
36.
BrownS., YoungH.K., and AmyesS.G.. 2005. Characterisation of OXA-51, a novel class D carbapenemase found in genetically unrelated clinical strains of Acinetobacter baumannii from Argentina. Clin. Microbiol. Infect. 11:15–23.
37.
HeritierC., PoirelL., FournierP.E., ClaverieJ.M., RaoultD., and NordmannP.. 2005. Characterization of the naturally occurring oxacillinase of Acinetobacter baumannii. Antimicrob. Agents Chemother. 49:4174–4179.
38.
HeritierC., PoirelL., LambertT., and NordmannP.. 2005. Contribution of acquired carbapenem-hydrolyzing oxacillinases to carbapenem resistance in Acinetobacter baumannii. Antimicrob. Agents Chemother. 49:3198–3202.
39.
RobledoI.E., AquinoE.E., SanteM.I., SantanaJ.L., OteroD.M., LeonC.F., and VazquezG.J.. 2010. Detection of KPC in Acinetobacter spp. in Puerto Rico. Antimicrob. Agents Chemother. 54:1354–1357.
40.
BonninR.A., NordmannP., PotronA., LecuyerH., ZaharJ.R., and PoirelL.. 2011. Carbapenem-hydrolyzing GES-type extended-spectrum beta-lactamase in Acinetobacter baumannii. Antimicrob. Agents Chemother. 55:349–354.
41.
BonninR.A., PoirelL., NaasT., PirsM., SemeK., SchrenzelJ., and NordmannP.. 2012. Dissemination of New Delhi metallo-beta-lactamase-1-producing Acinetobacter baumannii in Europe. Clin. Microbiol. Infect. 18:E362–E365.
42.
BoulangerA., NaasT., FortineauN., FigueiredoS., and NordmannP.. 2012. NDM-1-producing Acinetobacter baumannii from Algeria. Antimicrob. Agents Chemother. 56:2214–2215.
43.
FigueiredoS., PoirelL., PapaA., KoulouridaV., and NordmannP.. 2008. First identification of VIM-4 metallo-beta-lactamase in Acinetobacter spp. Clin. Microbiol. Infect. 14:289–290.
44.
RiccioM.L., FranceschiniN., BoschiL., CaravelliB., CornagliaG., FontanaR., AmicosanteG., and RossoliniG.M.. 2000. Characterization of the metallo-beta-lactamase determinant of Acinetobacter baumannii AC-54/97 reveals the existence of bla(IMP) allelic variants carried by gene cassettes of different phylogeny. Antimicrob. Agents Chemother. 44:1229–1235.
45.
AlkasabyN.M., and El Sayed ZakiM.. 2017. Molecular study of Acinetobacter baumannii isolates for metallo-beta-lactamases and extended-spectrum-beta-lactamases genes in intensive care unit, Mansoura University Hospital, Egypt. Int. J. Microbiol. 2017:3925868.
46.
RoutrayA., LavanyaP., SoniyaR., and MadhavanR.. 2013. Multiplex PCR for genes encoding prevalent OXA and NDM-1 carbapenemases in Acinetobacter. J. Pharm. Res. 7:324–326.
47.
HuangG., PengY., YangY., TangC., and FuY.. 2017. Multilocus sequence typing and molecular characterization of β-lactamase genes among Acinetobacter baumannii isolates in a burn center. Burns. 43:1473–;1478.
48.
KuoS.C., HuangW.C., HuangT.W., WangH.Y., LaiJ.F., ChenT.L., and LauderdaleT.L.. 2018. Molecular epidemiology of emerging blaOXA-23-like- and blaOXA-24-like-carrying Acinetobacter baumannii in Taiwan. Antimicrob. Agents Chemother. 62:e01215–01217.
49.
MohanamL., and MenonT.. 2017. Coexistence of metallo-beta-lactamase-encoding genes in Pseudomonas aeruginosa. Indian J. Med. Res. 146:S46–S52.
50.
DavoodiS., BoroumandM.A., SepehrisereshtS., and PourgholiL.. 2015. Detection of VIM-and IMP-type metallo-beta-lactamase genes in Acinetobacter baumannii isolates from patients in two hospitals in Tehran. Iran. J. Biotechnol. 13:63.
51.
GomaaF.A.M., HelalZ.H., and KhanM.I.. 2017. High prevalence of blaNDM-1, blaVIM, qacE, and qacEΔ1 genes and their association with decreased susceptibility to antibiotics and common hospital biocides in clinical isolates of Acinetobacter baumannii. Microorganisms. 5:18.
52.
CorvecS., PoirelL., NaasT., DrugeonH., and NordmannP.. 2007. Genetics and expression of the carbapenem-hydrolyzing oxacillinase gene blaOXA-23 in Acinetobacter baumannii. Antimicrob. Agents Chemother. 51:1530–1533.
53.
SegalH., JacobsonR.K., GarnyS., BamfordC.M., and ElishaB.G.. 2007. Extended -10 promoter in ISAba-1 upstream of blaOXA-23 from Acinetobacter baumannii. Antimicrob. Agents Chemother. 51:3040–3041.
54.
LeeY., KimC.-K., LeeH., JeongS.H., YongD., and LeeK.. 2011. A novel insertion sequence, ISAba10, inserted into ISAba1 adjacent to the bla OXA-23 gene and disrupting the outer membrane protein gene carO in Acinetobacter baumannii. Antimicrob. Agents Chemother. 55:361–363.
55.
MussiM.A., LimanskyA.S., and VialeA.M.. 2005. Acquisition of resistance to carbapenems in multidrug-resistant clinical strains of Acinetobacter baumannii: natural insertional inactivation of a gene encoding a member of a novel family of beta-barrel outer membrane proteins. Antimicrob. Agents Chemother. 49:1432–1440.
56.
GribunA., NitzanY., PechatnikovI., HershkovitsG., and KatcoffD.J.. 2003. Molecular and structural characterization of the HMP-AB gene encoding a pore-forming protein from a clinical isolate of Acinetobacter baumannii. Curr. Microbiol. 47:434–443.
57.
Catel-FerreiraM., NehmeR., MolleV., ArandaJ., BouffartiguesE., ChevalierS., BouG., JouenneT., and DeE.. 2012. Deciphering the function of the outer membrane protein OprD homologue of Acinetobacter baumannii. Antimicrob. Agents Chemother. 56:3826–3832.
58.
ChenF., WangL., WangM., XieY., XiaX., LiX., LiuY., CaoW., ZhangT., and LiP.. 2018. Genetic characterization and in vitro activity of antimicrobial combinations of multidrug-resistant Acinetobacter baumannii from a general hospital in China. Oncol. Lett. 15:2305–2315.
59.
TownerK., LeviK., VlassiadiM., and GroupA.S.. 2008. Genetic diversity of carbapenem-resistant isolates of Acinetobacter baumannii in Europe. Clin. Microbiol. Infect. 14:161–167.
60.
AkhiM.T., KhaliliY., GhotaslouR., YousefiS., KafilH.S., NaghiliB., and SheikhalizadehV.. 2018. Evaluation of carbapenem resistance mechanisms and its association with Pseudomonas aeruginosa infections in the Northwest of Iran. Microb. Drug Resist. 24:126–135.
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