Restricted accessResearch articleFirst published online 2022-09
Fecal Carriage of Extended-Spectrum β-Lactamases and pAmpC Producing Enterobacterales in an Iranian Community: Prevalence,Risk Factors,Molecular Epidemiology,and Antibiotic Resistance
This study aimed to determine the prevalence and risk factors associated with intestinal carriage of extended-spectrum β-lactamases producing Enterobacterales (ESBL-PE) and plasmid-mediated AmpC β-lactamase producing Enterobacterales (AmpC-PE) in healthy children in Ardabil, Iran. A total of 305 fecal samples were collected. Isolates underwent antimicrobial susceptibility testing, phenotypic and genotypic identification of β-lactamase production, and epidemiologic molecular typing. In total, 21.5%, 1.5%, and 1.2% of volunteers were extended-spectrum β-lactamase (ESBL)-, AmpC-, and simultaneous ESBL/AmpC-PE carriers, respectively. Escherichia coli was the predominant ESBL producing bacterium (70.2%) found in ESBL-PE colonized subjects. Beyond ESBL positive isolates, blaCTX-M group genes were the most common type (75.6%) and blaTEM (non-blaTEM-1 and non- blaTEM-2) were in the second place (25.6%). Among blaCTX-M genes, blaCTX-M-1 (55.3%) and blaCTX-M-15 (55.3%) were the most predominant types with equal prevalence. Some isolates were multi-enzyme producers. blaCIT and blaDHA genes were common AmpC type enzyme encoding genes found in AmpC-PE isolates. Most isolates produced both enzymes at the same time. The number of students in the classes was statistically associated with ESBL-PE intestinal carriage (p < 0.05). Moreover, 46 (65.7%), 3 (60%), 4 (100%), and 98 (39.8%) ESBL-, AmpC-, ESBL/AmpC, and non-ESBL/AmpC-PE isolates were multidrug-resistant, respectively. Overall, regardless of β-lactam antibiotics, 62% and 59.5% of isolates were resistant to co-trimoxazole and tetracycline, respectively. The majority of ESBL producing E. coli isolates (69.2%) belonged to phylogroup A. According to Enterobacterial repetitive intergenic consensus polymerase chain reaction, there was no clonal relatedness between isolates. This study showed a high rate of multi-resistant ESBL-PE intestinal carriage among healthy individuals in Iran.
Get full access to this article
View all access options for this article.
References
1.
MurrayP.R., RosenthalK.S, Pfaller M.A(eds). 2016. Medical microbiology, 8th ed. Elsevier, Philadelphia, pp. 251–264.
2.
MansouriS., and AbbasiS.. 2010. Prevalence of multiple drug resistant clinical isolates of extended-spectrum β-lactamase producing Enterobacteriaceae in southeast Iran. Iran J. Med. Sci. 35: 101–108.
3.
PelegA.Y., and HooperD.C.. 2010. Hospital acquired infections due to gram negative bacteria. N. Engl. J. Med. 362:1804–1813.
4.
ArzanlouM., ChaiW.C, and VenterH.. 2017. Intrinsic, adaptive and acquired antimicrobial resistance in gram negative bacteria, Essays Biochem. 61:49–59.
5.
RuhE., J.Zakka, K.Hoti, et al. 2019. Extended spectrum β-lactamase, plasmid-mediated AmpC β-lactamase, fluoroquinolone resistance, and decreased susceptibility to carbapenems in Enterobacteriacae: fecal carriage rates and associated risk factors in the community of northern Cyprus. Antimicrob. Resist. Infect. Control. 8:98.
6.
AraqueM., and LabradorI.. 2018. Prevalence of fecal carriage of CTX-M-15 β-lactamase producing Escherichia coli in healthy children from a rural Andean village in Venezuela. Osong. Public Health Res. Perspect. 9:9–15.
7.
MosesA., F.Bwanga, Y.Boum, and BaziraJ.. 2014. Prevalence and genotypic characterization of extended spectrum β-lactamases produced by gram negative bacilli at a tertiary care hospital in rural south western Uganda. Br. Microbiol. Res. J. 4:1541–1550.
8.
GhafourianS., SadeghifardN., SoheiliS., and SekawiZ.. 2015. Extended spectrum β-lactamases: definition, classification and epidemiology. Curr. Issues Mol. Biol. 17:11–21.
9.
NaasT., PoirelL., and NordmannP.. 2008. Minor extended spectrum β-lactamases. Clin. Microbiol .Infect. 14,(Suppl. 1)::42–52.
10.
CastanheiraM., SimnerP.J, and BradfordP.A.. 2021. Extended spectrum β-lactamases: an update on their characteristics, epidemiology and detection. JAC Antimicrob. Resist. 16:3.
11.
Pérez-PérezF.J., and HansonN.D.. 2002. Detection of plasmid-mediated AmpC β-lactamase genes in clinical isolates by using multiplex PCR. J. Clin. Microbiol. 40:2153–2162.
12.
KawamuraK., N.Nagano, M.Suzuki, Wachino Ji, K.Kimura, and ArakawaY.. 2017. ESBL-producing Escherichia coli and its rapid rise among healthy people. Food Saf. 5:122–150.
13.
WoertherP.L., C.Burdet, E.Chachaty, and AndremontA.. 2013. Trends in human fecal carriage of extended spectrum β-lactamases in the community: toward the globalization of CTX-M. Clin. Microbiol. Rev. 26:744–758.
14.
KammiliN., N.Cherukuri, PalvaiS, et al.2014. Molecular epidemiology of extended spectrum β-lactamase producing Escherichia coli and Klebsiella pneumoniae in a tertiary care hospital. Indian J. Med. Microbiol. 32:205–207.
15.
RashidM., ModiS., SarwatT., ChanderY., RastogiV., and ManochaH.. 2015. Carriage of ESBL and AmpC-positive Enterobacteriacae in gastrointestinal tract of healthy community subjects and hospitalized patients and detection of blaCTX-M gene in ESBL positive isolates. Ind. Med. Gaz. 149: 212–218.
16.
KaarmeJ., RiedelH., SchaalW., YinH., NeveusT., and MelhusA.. 2018. Rapid increase in carriage rates of Enterobacteriaceae producing extended spectrum β-lactamases in healthy preschool children, Sweden. Emerg. Infect. Dis. 24:1874–1881.
17.
MahonC., and Lehman D.(eds). 2015. Textbook of diagnostic microbiology, 5th ed. Saunders, Philadelphia,, PA, pp. 420–454.
18.
Clinical and Laboratory Standards Institute (CLSI). 2022. Performance Standards for Antimicrobial Susceptibility Testing, 32nd ed. CLSI supplement M100 (ISBN 978-1-68440-134-5 [Print]; ISBN 978-1-68440-135-2 [Electronic]). Clinical and Laboratory Standards, Institute, USA.
19.
SchwalbeR., Steele-MooreL., and Goodwin A.C(eds). 2007. Antimicrobial susceptibility testing protocols, 1st ed. CRC Press, New York.
20.
The European Committee on Antimicrobial Susceptibility Testing. 2022. Breakpoint tables for interpretation of MICs and zone diameters. Version 12.0. Available at www.eucast.org
21.
PoulouA., GrivakouE., VrioniG., et al.2014. Modified CLSI extended spectrum β-lactamase (ESBL) confirmatory test for phenotypic detection of ESBLs among Enterobacteriacae producing various β-lactamases. J. Clin. Microbiol. 52:1483–1489.
22.
GuptaG., TakV., and MathurP.. 2014. Detection of AmpC β-lactamases in gram-negative bacteria. J. Lab. Phys. 6:1–6.
23.
RuppéE., HemS., LathS., et al.2009. CTX-M β-lactamases in Escherichia coli from community acquired urinary tract infections, Cambodia. Emerg. Infect. Dis. 15:741–748.
24.
SaladinM., CaoV.T, LambertT., et al.2002. Diversity of CTX-M β-lactamases and their promoter regions from Enterobacteriaceae isolated in three Parisian hospitals. FEMS Microbiol. Lett. 209:161–168.
25.
ArletG., RouveauM., and PhilipponA.. 1997. Substitution of alanine for aspartate at position 179 in the SHV-6 extended spectrum β-lactamase. FEMS Microbiol. Lett. 152:163–167.
26.
LavollayM., MamloukK., FrankT., et al. 2006. Clonal dissemination of a CTX-M-15 β-lactamase producing Escherichia coli strain in the Paris area, Tunis, and Bangui. Antimicrob. Agents Chemother. 50:2433–2438.
27.
StewardC.D., RasheedJ.K, HubertS.K, et al.2001. Characterization of clinical isolates of Klebsiella pneumoniae from 19 laboratories using the National Committee for Clinical Laboratory Standards extended spectrum β-lactamase detection methods. J. Clin. Microbiol. 39:2864–2872.
28.
ChmelnitskyI., CarmeliY., LeavittA., SchwaberM.J, and Navon-VeneziaS.. 2005. CTX-M-2 and a new CTX-M-39 enzyme are the major extended spectrum β-lactamases in multiple Escherichia coli clones isolated in Tel Aviv, Israel. Antimicrob. Agents Chemother. 49:4745–4750.
29.
HuangS., DaiW., SunS., ZhangX., and ZhangL.. 2012. Prevalence of plasmid-mediated quinolone resistance and aminoglycoside resistance determinants among carbapeneme non-susceptible Enterobacter cloacae. PLoS One. 7:e47636.
30.
MendonçaN., LeitãoJ., ManageiroV., FerreiraE., and CaniçaM.. 2007. Spread of extended spectrum β-lactamase CTX-M producing Escherichia coli clinical isolates in community and nosocomial environments in Portugal. Antimicrob Agents Chemother. 51:1946–1955.
31.
ClermontO., DhanjiH., UptonM., et al.2009. Rapid detection of the O25b-ST131 clone of Escherichia coli encompassing the CTX-M-15 producing strains. J. Antimicrob. Chemother. 64:274–277.
32.
ClermontO., LavollayM., VimontS., et al.2008. The CTX-M-15 producing Escherichia coli diffusing clone belongs to a highly virulent B2 phylogenetic subgroup. J. Antimicrob. Chemother. 61:1024–1028.
33.
LavillaS., González-LópezJ.J, SabatéM., et al.2007. Prevalence of qnr genes among extended spectrum β-lactamase producing enterobacterial isolates in Barcelona, Spain. J. Antimicrob. Chemother. 61:291–295.
34.
MomtazH., KarimianA., MadaniM., et al.2013. Uropathogenic Escherichia coli in Iran: serogroup distributions, virulence factors and antimicrobial resistance properties. Ann. Clin. Microbiol. Antimicrob. 12:8.
35.
ClermontO., ChristensonJ.K, DenamurE., and GordonD.M.. 2013. The Clermont Escherichia coli phylo-typing method revisited: improvement of specificity and detection of new phylo-groups. Environ. Microbiol. Rep. 5:58–65.
36.
LescatM., ClermontO., WoertherP.L, et al.2013. Commensal Escherichia coli strains in Guiana reveal a high genetic diversity with host-dependent population structure. Environ. Microbiol. Rep. 5:49–57.
37.
VersalovicJ., KoeuthT., and LupskiJ.R.. 1991. Distribution of repetitive DNA sequences in eubacteria and application to fingerprinting of bacterial genomes. Nucleic Acids Res. 19:6823–6831.
38.
GhotaslouR., SadeghiM.R, AkhiM.T, HasaniA., and AsgharzadehM.. 2018. Prevalence and antimicrobial susceptibility patterns of ESBL, Ampc and carbapenemase-producing Enterobactericeae isolated from hospitalized patients in Azerbaijan, Iran. Iran J. Pharm. Res. 17(Suppl.), 79–88.
39.
KayasthaK., DhungelB., KarkiS., et al. 2020. Extended spectrum β-lactamase producing Escherichia coli and Klebsiella species in pediatric patients visiting international friendship children's hospital, Kathmandu, Nepal. Infect. Dis. (Auckl). 13:1178633720909798.
40.
Mughini-GrasL., Dorado-GarcíaA., van DuijkerenE., et al.2019. Attributable sources of community-acquired carriage of Escherichia coli containing β-lactam antibiotic resistance genes: a population-based modelling study. Lancet Planet Health. 3: e357–e369.
41.
YousefipourM., RasoulinejadM., HadadiA., et al.2019. Bacteria producing extended spectrum β-lactamases (ESBLs) in hospitalized patients: prevalence, antimicrobial resistance pattern and its main determinants. Iran. J. Pathol. 14:61–67.
42.
Abdul RahmanE.M., and El-SherifR.H.. 2011. High rates of intestinal colonization with extended spectrum β-lactamase producing Enterobacteriacae among healthy individuals. J. Investig. Med. 59:1284–1286.
43.
Rubee ChanuT., ShahP.K, SoniS., and GhoshA.N.. 2019. Phenotypic detection of extended spectrum, AmpC, metallo β-lactamases and their co-existence in clinical isolates of commonly isolated gram negative bacteria in GKGH hospital, Bhuj. Int. J. Med. Microbiol. Trop Dis. 5:52–56.
44.
BatistaA.D., RodriguesD., FigueirasA., Zapata-CachafeiroM., RoqueF., and HerdeiroM.T.. 2020. Antibiotic dispensation without a prescription worldwide: a systematic review. Antibiotics. 9:786.
45.
AbbasianH., HajimolaaliM., YektadoostA., and ZartabS.. 2019. Antibiotic utilization in Iran 2000–2016: pattern analysis and benchmarking with organization for economic co-operation and development countries. J. Res. Pharm. Pract. 8:162–167.
46.
OuédraogoA.S., SanouS., KissouA., et al.2017. Fecal carriage of Enterobacteriacae producing extended spectrum β-lactamases in hospitalized patients and healthy community volunteers in Burkina Faso. Microb. Drug Resist. 23:63–70.
47.
KizilatesF., YakupogullariY., BerkH., OztoprakN., and OtluB.. 2020. Risk factors for fecal carriage of extended spectrum β-lactamase producing and carbapenem-resistant Escherichia coli and Klebsiella pneumonia strains among patients at hospital admission. Am. J. Infect. Control. S0196-65533063-X.
48.
ReulandE.A., Al NaiemiN., KaiserA.M, et al. 2016. Prevalence and risk factors for carriage of ESBL-producing Enterobacteriacae in Amsterdam. J. Antimicrob. Chemother. 71:1076–1082.
49.
LonchelC.M., MeexC., Gangoue-PiebojiJ., et al.2012. Proportion of extended spectrum β-lactamase producing Enterobacteriacae in community setting in Ngaoundere, Cameron. BMC Infect. Dis. 12:53.
50.
JabalameliL., BeigverdiR., RanjbarH.H, PouriranR., JabalameliF., and EmaneiniM.. 2021. Phenotypic and genotypic prevalence of extended spectrum β-lactamase producing Escherichia coli: a systematic review and meta-analysis in Iran. Microb. Drug. Resist. 27:73–86.
51.
NiumsupP.R., TansawaiU., Na-UdomA., et al.2018. Prevalence and risk factors for intestinal carriage of CTX-M type ESBLs in Enterobacteriacae from a Thai community. Eur. J. Clin. Microbiol. Infect. Dis. 37:69–75.
52.
GajamerV.R., BhattacharjeeA., PaulD., et al.2020. High prevalence of carbapenemase, AmpC β-lactamase and aminoglycoside resistance genes in extended spectrum β-lactamase-positive uropathogens from Northern India. J. Glob. Antimicrob. Resist. 20:197–203.
53.
IsendahlJ., Turlej-RogackaA., ManjubaC., RodriguesA., GiskeC.G, and NauclérP.. 2012. Fecal carriage of ESBL-producing E. coli and K. pneumonia in children in Guinea-Bissau: a hospital-based cross-sectional study. PLoS One. 7:e51981.
54.
D'AngeloR.G., JohnsonJ.K, BorkJ.T, and HeilE.L.. 2016. Treatment options for extended spectrum β-lactamase (ESBL) and AmpC-producing bacteria. Expert. Opin. Pharmacother. 17:953–967.
55.
AfzaliH., FiroozehF., AmiriA., MoniriR., and ZibaeiM.. 2015. Characterization of CTX-M type extended spectrum β-lactamase producing Klebsiella spp in Kashan, Iran. Jundishapur J. Microbiol. 8:1–5.
56.
ZhangH., ZhouY., GuoS., and ChangW.. 2015. High prevalence and risk factors of fecal carriage of CTX-M type extended spectrum β-lactamase producing Enterobacteriacae from healthy rural residents of Taian, China. Front. Microbiol. 6:239.
57.
MirzaeeM., OwliaP., and MansouriS.. 2009. Distribution of CTX-M β-lactamase genes among Escherichia coli strains isolated from patients in Iran. Lab. Med. 40:724–727.
58.
PoirelL., GniadkowskiM., and NordmannP.. 2002. Biochemical analysis of the ceftazidime-hydrolyzing extended spectrum β-lactamase CTX-M-15 and of its structurally related β-lactamase CTX-M-3. J. Antimicrob. Chemother. 50:1031–1034.
59.
LiuX.Q., and LiuY.R.. 2016. Detection and genotype analysis of AmpC β-lactamase in Klebsiella pneumonia from tertiary hospitals. Exp. Ther. Med. 12:480–484.
60.
HuangI.F., LeeW.Y, WangJ.L, et al.2018. Fecal carriage of multidrug-resistant Escherichia coli by community children in southern Taiwan. BMC Gastroenterol. 18:86.
61.
Ouchar MahamatO., TidjaniA., LounnasM., et al.2019. Fecal carriage of extended spectrum β-lactamase-producing Enterobacteriaceae in hospital and community settings in Chad. Antimicrob. Resist. Infect. Control. 8:169.
62.
PatersonD.L., and BonomoR.A.. 2005. Extended-spectrum beta-lactamases: a clinical update. Clin. Microbiol. Rev. 18:657–686.
63.
OnduruO.G., AboudS., NyirendaT.S, RumishaS.F, and MkakosyaR.S.. 2021. Antimicrobial susceptibility testing profiles of ESBL-producing Enterobacterales isolated from hospital and community adult patients in Blantyre, Malawi. IJID. Reg. 6:47–52.
64.
GordonD.M., and CowlingA.. 2003. The distribution and genetic structure of Escherichia coli in Australian vertebrates: host and geographic effects. Microbiology. 149:3575–3586.
65.
TenaillonO., SkurnikD., PicardB., and DenamurE.. 2010. The population genetics of commensal Escherichia coli. Nat. Rev. Microbiol. 8:207–217.
66.
Stoppe, N.d.C., SilvaJ.S, CarlosC., et al. 2017. Worldwide phylogenetic group patterns of Escherichia coli from commensal human and wastewater treatment plant isolates. Front. Microbiol. 8:2512.
Supplementary Material
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
