Brazilian Journal of Microbiology

, Volume 50, Issue 1, pp 175–183 | Cite as

Characterization of multidrug-resistant Gram-negative bacilli isolated from hospitals effluents: first report of a blaOXA-48-like in Klebsiella oxytoca, Algeria

  • Khadidja Yousfi
  • Abdelaziz Touati
  • Brigitte Lefebvre
  • Philippe Garneau
  • Soumia Brahmi
  • Alima Gharout-Sait
  • Josée Harel
  • Sadjia BekalEmail author
Environmental Microbiology - Research Paper


The antibiotic susceptibility profile and antimicrobial resistance determinants were characterized on Gram-negative bacilli (GNB) isolated from Algerian hospital effluents. Among the 94 isolates, Enterobacteriaceae was the predominant family, with Escherichia coli and Klebsiella pneumoniae being the most isolated species. In non-Enterobacteriaceae, Acinetobacter and Aeromonas were the predominant species followed by Pseudomonas, Comamonas, Pasteurella, and Shewanella spp. The majority of the isolates were multidrug-resistant (MDR) and carried different antimicrobial resistance genes including blaCTX-M, blaTEM, blaSHV, blaOXA-48-like, blaOXA-23, blaOXA-51, qnrB, qnrS, tet(A), tet(B), tet(C), dfrA1, aac(3)-IIc (aacC2), aac(6′)-1b, sul1, and sul2. The qacEΔ1-sul1 and intI2 signatures of class 1 and class 2 integrons, respectively, were also detected. Microarray hybridization on MDR E. coli revealed additional resistance genes (aadA1 and aph3strA, tet30, mphA, dfrA12, blacmy2, blaROB1, and cmlA1) and classified the tested strains as commensals, thus highlighting the potential role of humans in antibiotic resistance dissemination. This study is the first report of blaOXA-48-like in Klebsiella oxytoca in Algeria and blaOXA-23 in A. baumannii in Algerian hospital effluents. The presence of these bacteria and resistance genes in hospital effluents represents a serious public health concern since they can be disseminated in the environment and can colonize other hosts.


Multidrug resistance Hospital effluents Gram-negative bacilli Algeria 



The authors would like to thank Nancy Cloutier, Isabelle Robillard, Simon Wang, Karine Desjardins, Dominique Paquette, Annie Vezina, and François Robillard for their Lab assistance and Dr. Valentine Usongo for reviewing the manuscript.


  1. 1.
    Biyela PT, Lin J, Bezuidenhout CC (2004) The role of aquatic ecosystems as reservoirs of antibiotic resistant bacteria and antibiotic resistance genes. Water Sci Technol 50(1):45–50CrossRefGoogle Scholar
  2. 2.
    Kümmerer K (2004) Resistance in the environment. J Antimicrob Chemother 54(2):311–320CrossRefGoogle Scholar
  3. 3.
    Korzeniewska E, Harnisz M (2013) Beta-lactamase-producing Enterobacteriaceae in hospital effluents. J Environ Manag 123:1–7CrossRefGoogle Scholar
  4. 4.
    Pauwels B, Verstraete W (2006) The treatment of hospital wastewater: an appraisal. J Water Health 4(4):405–416CrossRefGoogle Scholar
  5. 5.
    Anssour L, Messai Y, Derkaoui M, Alouache S, Estepa V, Somalo S, Torres C, Bakour R (2014) ESBL, plasmidic AmpC, and associated quinolone resistance determinants in coliforms isolated from hospital effluent: first report of qnrB2, qnrB9, qnrB19, and bla CMY-4 in Algeria. J Chemother 26(2):74–79CrossRefGoogle Scholar
  6. 6.
    Yousfi K, Touati A, Lefebvre B, Fournier É, Côté JC, Soualhine H, Walker M, Bougdour D, Tremblay C, Bekal S (2016) A novel plasmid, pSx1, harboring a new Tn1696 derivative from extensively drug-resistant Shewanella xiamenensis encoding OXA-416. Microb Drug Resist 23(4):429–436CrossRefGoogle Scholar
  7. 7.
    Farmer JJ, Davis BR, Hickman-Brenner FW et al (1985) Biochemical identification of new species and biogroups of Enterobacteriaceae isolated from clinical specimens. J Clin Microbiol 21(1):46–76Google Scholar
  8. 8.
    Clinical and Laboratory Standards Institute (CLSI) (2014) Performance standards for antimicrobial susceptibility testing. Twenty-Fourth Informational Supplement M100-S24. CLSI, WayneGoogle Scholar
  9. 9.
    Rasheed JK, Jay C, Metchock B, Berkowitz F, Weigel L, Crellin J, Steward C, Hill B, Medeiros AA, Tenover FC (1997) Evolution of extended-spectrum beta-lactam resistance (SHV-8) in a strain of Escherichia coli during multiple episodes of bacteremia. Antimicrob Agents Chemother 41(3):647–653CrossRefGoogle Scholar
  10. 10.
    Monstein HJ, Ostholm-Balkhed A, Nilsson MV, Nilsson M, Dornbusch K, Nilsson LE (2007) Multiplex PCR amplification assay for the detection of bla SHV, bla TEM and bla CTX-M genes in Enterobacteriaceae. APMIS 115(12):1400–1408CrossRefGoogle Scholar
  11. 11.
    Woodford N, Ellington MJ, Coelho JM et al (2006) Multiplex PCR for genes encoding prevalent OXA carbapenemases in Acinetobacter spp. Int J Antimicrob Agents 27(4):351–353CrossRefGoogle Scholar
  12. 12.
    Brown S, Young HK, Amyes SG (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(1):15–23CrossRefGoogle Scholar
  13. 13.
    Cattoir V, Poirel L, Rotimi V, Soussy CJ, Nordmann P (2007) Multiplex PCR for detection of plasmid-mediated quinolone resistance qnr genes in ESBL-producing enterobacterial isolates. J Antimicrob Chemother 60(2):394–397CrossRefGoogle Scholar
  14. 14.
    Robicsek A, Strahilevitz J, Sahm DF, Jacoby GA, Hooper DC (2006) qnr prevalence in ceftazidime-resistant Enterobacteriaceae isolates from the United States. Antimicrob Agents Chemother 50(8):2872–2874CrossRefGoogle Scholar
  15. 15.
    Park CH, Robicsek A, Jacoby GA, Sahm D, Hooper DC (2006) Prevalence in the United States of aac(6′)-Ib-cr encoding a ciprofloxacin-modifying enzyme. Antimicrob Agents Chemother 50(11):3953–3955CrossRefGoogle Scholar
  16. 16.
    Gow SP, Waldner CL, Harel J, Boerlin P (2008) Associations between antimicrobial resistance genes in fecal generic Escherichia coli isolates from cow-calf herds in western Canada. Appl Environ Microbiol 74(12):3658–3666CrossRefGoogle Scholar
  17. 17.
    Maynard C, Fairbrother JM, Bekal S, Sanschagrin F, Levesque RC, Brousseau R, Masson L, Lariviere S, Harel J (2003) Antimicrobial resistance genes in enterotoxigenic Escherichia coli O149:K91 isolates obtained over a 23-year period from pigs. Antimicrob Agents Chemother 47(10):3214–3221CrossRefGoogle Scholar
  18. 18.
    Ploy MC, Denis F, Courvalin P, Lambert T (2000) Molecular characterization of integrons in Acinetobacter baumannii: description of a hybrid class 2 integron. Antimicrob Agents Chemother 44(10):2684–2688CrossRefGoogle Scholar
  19. 19.
    Jakobsen L, Garneau P, Kurbasic A et al (2011) Microarray-based detection of extended virulence and antimicrobial resistance gene profiles in phylogroup B2 Escherichia coli of human, meat and animal origin. J Med Microbiol 60:1502–1511CrossRefGoogle Scholar
  20. 20.
    Bonnet C, Diarrassouba F, Brousseau R, Masson L, Topp E, Diarra MS (2009) Pathotype and antibiotic resistance gene distributions of Escherichia coli isolates from broiler chickens raised on antimicrobial-supplemented diets. Appl Environ Microbiol 75(22):6955–6962CrossRefGoogle Scholar
  21. 21.
    Clermont O, Bonacorsi S, Bingen E (2000) Rapid and simple determination of the Escherichia coli phylogenetic group. Appl Environ Microbiol 66(10):4555–4558CrossRefGoogle Scholar
  22. 22.
    Chapman TA, Wu XY, Barchia I, Bettelheim KA, Driesen S, Trott D, Wilson M, Chin JJC (2006) Comparison of virulence gene profiles of Escherichia coli strains isolated from healthy and diarrheic swine. Appl Environ Microbiol 72(7):4782–4795CrossRefGoogle Scholar
  23. 23.
    Lau HY, Huffnagle GB, Moore TA (2008) Host and microbiota factors that control Klebsiella pneumoniae mucosal colonization in mice. Microbes Infect 10(12–13):1283–1290CrossRefGoogle Scholar
  24. 24.
    Allocati N, Masulli M, Alexeyev MF, Di Ilio C (2013) Escherichia coli in Europe: an overview. Int J Environ Res Public Health 10(12):6235–6254CrossRefGoogle Scholar
  25. 25.
    Hrenovic J, Durn G, Goic-Barisic I, Kovacic A (2014) Occurrence of an environmental Acinetobacter baumannii strain similar to a clinical isolate in paleosol from Croatia. Appl Environ Microbiol 80(9):2860–2866CrossRefGoogle Scholar
  26. 26.
    Chen PL, Wu CJ, Tsai PJ, Tang HJ, Chuang YC, Lee NY, Lee CC, Li CW, Li MC, Chen CC, Tsai HW, Ou CC, Chen CS, Ko WC (2014) Virulence diversity among bacteremic Aeromonas isolates: ex vivo, animal, and clinical evidences. PLoS One 9(issue 11):e111213CrossRefGoogle Scholar
  27. 27.
    Partridge SR (2011) Analysis of antibiotic resistance regions in Gram-negative bacteria. FEMS Microbiol Rev 35(5):820–855CrossRefGoogle Scholar
  28. 28.
    Canton R, Gonzalez-Alba JM, Galan JC (2012) CTX-M enzymes: origin and diffusion. Front Microbiol 3:110CrossRefGoogle Scholar
  29. 29.
    Gharout-Sait A, Touati A, Benallaaoua S et al (2012) CTX-M from community-acquired urinary tract, Algeria. Afr J Microbiol Res 6(25):5306–5313Google Scholar
  30. 30.
    Brahmi S, Dunyach-Remy C, Touati A, Lavigne JP (2015) CTX-M-15-producing Escherichia coli and the pandemic clone O25b-ST131 isolated from wild fish in Mediterranean Sea. Clin Microbiol Infect 21(3):e18–e20CrossRefGoogle Scholar
  31. 31.
    Marinescu F, Marutescu L, Savin I, Lazar V (2015) Antibiotic resistance markers among Gram-negative isolates from wastewater and receiving rivers in South Romania. Rom Biotechnol Lett 20(1):10055–10069Google Scholar
  32. 32.
    Maragakis LL, Perl TM (2008) Acinetobacter baumannii: epidemiology, antimicrobial resistance, and treatment options. Clin Infect Dis 46(8):1254–1263CrossRefGoogle Scholar
  33. 33.
    Bakour S, Touati A, Bachiri T, Sahli F, Tiouit D, Naim M, Azouaou M, Rolain JM (2014) First report of 16S rRNA methylase ArmA-producing Acinetobacter baumannii and rapid spread of metallo-beta-lactamase NDM-1 in Algerian hospitals. J Infect Chemother 20(issue 11):696–701CrossRefGoogle Scholar
  34. 34.
    Brahmi S, Touati A, Cadiere A et al (2016) First description of two sequence type 2 Acinetobacter baumannii isolates carrying OXA-23 carbapenemase in Pagellus acarne fished from the Mediterranean Sea near Bejaia, Algeria. Antimicrob Agents Chemother 60(4):2513–2515CrossRefGoogle Scholar
  35. 35.
    Loucif L, Kassah-Laouar A, Saidi M, Messala A, Chelaghma W, Rolain JM (2016) Outbreak of OXA-48-producing Klebsiella pneumoniae involving a sequence type 101 clone in Batna university hospital, Algeria. Antimicrob Agents Chemother 60(12):7494–7497Google Scholar
  36. 36.
    Baba Ahmed-Kazi Tani Z, Arlet G (2014) News of antibiotic resistance among Gram-negative bacilli in Algeria. Pathol Biol 62(3):169–178CrossRefGoogle Scholar
  37. 37.
    Guo X, Xia R, Han N, Xu H (2011) Genetic diversity analyses of class 1 integrons and their associated antimicrobial resistance genes in Enterobacteriaceae strains recovered from aquatic habitats in China. Lett Appl Microbiol 52(6):667–675CrossRefGoogle Scholar
  38. 38.
    Anssour L, Messai Y, Estepa V, Torres C, Bakour R (2016) Characteristics of ciprofloxacin-resistant Enterobacteriaceae isolates recovered from wastewater of an Algerian hospital. J Infect Dev Ctries 10(7):728–734CrossRefGoogle Scholar
  39. 39.
    Horcajada JP, Soto S, Gajewski A, Smithson A, Jimenez de Anta MT, Mensa J, Vila J, Johnson JR (2005) Quinolone-resistant uropathogenic Escherichia coli strains from phylogenetic group B2 have fewer virulence factors than their susceptible counterparts. J Clin Microbiol 43(6):2962–2964CrossRefGoogle Scholar

Copyright information

© Sociedade Brasileira de Microbiologia 2018

Authors and Affiliations

  • Khadidja Yousfi
    • 1
    • 2
  • Abdelaziz Touati
    • 1
  • Brigitte Lefebvre
    • 2
  • Philippe Garneau
    • 3
  • Soumia Brahmi
    • 1
  • Alima Gharout-Sait
    • 1
  • Josée Harel
    • 3
  • Sadjia Bekal
    • 2
    • 4
    Email author
  1. 1.Laboratoire d’Écologie Microbienne, FSNVUniversité de BéjaiaBéjaiaAlgeria
  2. 2.Laboratoire de santé publique du QuébecInstitut national de santé publique du QuébecSainte-Anne-de-BellevueCanada
  3. 3.Faculté de Médecine Vétérinaire, Centre de Recherche en Infectiologie PorcineUniversité de MontréalSt-HyacintheCanada
  4. 4.Département de Microbiologie, Infectiologie et ImmunologieUniversité de MontréalMontréalCanada

Personalised recommendations