Applied Microbiology and Biotechnology

, Volume 97, Issue 1, pp 329–340 | Cite as

Diversity and antibiotic resistance of Acinetobacter spp. in water from the source to the tap

  • Carlos Narciso-da-Rocha
  • Ivone Vaz-Moreira
  • Liselott Svensson-Stadler
  • Edward R. B. Moore
  • Célia M. ManaiaEmail author
Applied microbial and cell physiology


Acinetobacter spp. are ubiquitous bacteria in the environment. Acinetobacter spp. isolated from a municipal drinking water treatment plant and from connected tap water were identified to the species level on the basis of rpoB gene partial sequence analysis. Intraspecies variation was assessed based on the analysis of partial sequences of housekeeping genes (rpoB, gyrB, and recA). Antibiotic resistance was characterized using the disk diffusion method and isolates were classified as wild or non-wild type (non-WT), according to the observed phenotype. The strains of Acinetobacter spp. were related to 11 different validly published species, although three groups of isolates, presenting low rpoB sequence similarities with previously described species, may represent new species. Most of the isolates were related to the species A. johnsonii and A. lwoffii. These two groups, as well as others related to the species A. parvus and A. tjernbergiae, were detected in the water treatment plant and in tap water. Other strains, related to the species A. pittii and A. beijerinckii, were isolated only from tap water. Most of the isolates (80 %) demonstrated wild type (WT) to all of the 12 antibiotics tested. Non-WT for tetracycline, meropenem, and ceftazidime, among others, were observed in water treatment plant or in tap water samples. Although, in general, this study suggests a low prevalence of acquired antibiotic resistance in water Acinetobacter spp., the potential of some species to acquire and disseminate resistance via drinking water is suggested.


Drinking water Antibiotic resistance ECOFF gyrrecrpo



The authors gratefully acknowledge the workers from the water treatment plant for their kind collaboration on sample collections and the people that kindly allowed the sampling of water from their taps.

This study was financed by Fundação para a Ciência e a Tecnologia (projects PTDC/AMB/70825/2006, PTDC/AAC-AMB/113840/2009 and IVM grant SFRH/BD/27978/2006). CNR was supported by ERASMUS placement agreement. LS-S and ERBM were supported, in part, by funding of the ALF-medel för forskning Västra Götaland Region project ALFGBG-11574 and the FoU-Västra Götaland Region project VGFOUREG-157801.


  1. Bartual SG, Seifert H, Hippler C, Luzon MA, Wisplinghoff H, Rodriguez-Valera F (2005) Development of a multilocus sequence typing scheme for characterization of clinical isolates of Acinetobacter baumannii. J Clin Microbiol 43:4382–4390CrossRefGoogle Scholar
  2. Bergogne-Berezin E, Towner KJ (1996) Acinetobacter spp. as nosocomial pathogens: microbiological, clinical, and epidemiological features. Clin Microbiol Rev 9:148–165Google Scholar
  3. Bhargava N, Sharma P, Capalash N (2010) Quorum sensing in Acinetobacter: an emerging pathogen. Crit Rev Microbiol 36:349–360CrossRefGoogle Scholar
  4. Dhakephalkar PK, Chopade BA (1994) High levels of multiple metal resistance and its correlation to antibiotic resistance in environmental isolates of Acinetobacter. Biometals 7:67–74CrossRefGoogle Scholar
  5. Dimopoulou ID, Kartali SI, Kartalis GN, Manolas KI, Simopoulos KE, Vargemezis BA, Theodoropoulou-Rodiou G, Bowler IC, Crook DW (2003) Relationship between nosocomial Acinetobacter species occurring in two geographical areas (Greece and the UK). J Hosp Infect 54:207–211CrossRefGoogle Scholar
  6. Eichler S, Christen R, Holtje C, Westphal P, Botel J, Brettar I, Mehling A, Hofle MG (2006) Composition and dynamics of bacterial communities of a drinking water supply system as assessed by RNA- and DNA-based 16S rRNA gene fingerprinting. Appl Environ Microbiol 72:1858–1872CrossRefGoogle Scholar
  7. Figueira V, Vaz-Moreira I, Silva M, Manaia CM (2011) Diversity and antibiotic resistance of Aeromonas spp. In drinking and waste water treatment plants. Water Res 45:5599–5611CrossRefGoogle Scholar
  8. Fishbain J, Peleg AY (2010) Treatment of Acinetobacter infections. Clin Infect Dis 51:79–84CrossRefGoogle Scholar
  9. Gonzalez CJ, Santos JA, Garcia-Lopez ML, Otero A (2000) Psychrobacters and related bacteria in freshwater fish. J Food Prot 63:315–321Google Scholar
  10. Gundi VA, Dijkshoorn L, Burignat S, Raoult D, La Scola B (2009) Validation of partial rpoB gene sequence analysis for the identification of clinically important and emerging Acinetobacter species. Microbiol 155:2333–2341CrossRefGoogle Scholar
  11. Hantsis-Zacharov E, Halpern M (2007) Culturable psychrotrophic bacterial communities in raw milk and their proteolytic and lipolytic traits. Appl Environ Microbiol 73:7162–7168CrossRefGoogle Scholar
  12. Henwood CJ, Gatward T, Warner M, James D, Stockdale MW, Spence RP, Towner KJ, Livermore DM, Woodford N (2002) Antibiotic resistance among clinical isolates of Acinetobacter in the UK, and in vitro evaluation of tigecycline (gar-936). J Antimicrob Chemother 49:479–487CrossRefGoogle Scholar
  13. Hoefel D, Monis PT, Grooby WL, Andrews S, Saint CP (2005) Profiling bacterial survival through a water treatment process and subsequent distribution system. J Appl Microbiol 99:175–186CrossRefGoogle Scholar
  14. Hong PY, Hwang C, Ling F, Andersen GL, LeChevallier MW, Liu WT (2010) Pyrosequencing analysis of bacterial biofilm communities in water meters of a drinking water distribution system. Appl Environ Microbiol 76:5631–5635CrossRefGoogle Scholar
  15. Idzenga D, Schouten MA, van Zanten AR (2006) Outbreak of Acinetobacter genomic species 3 in a Dutch intensive care unit. J Hosp Infect 63:485–487CrossRefGoogle Scholar
  16. Jukes TH, Cantor CR (1969) Evolution of protein molecules. In: Munro HN (ed) Mammalian protein metabolism, edn. Academic Press, New York, pp 21–132Google Scholar
  17. Juni E (2005) Proteobacteria, part b, the gammaproteobacteria. In: Brenner DJ, Krieg NR, Stanley JT (eds) Bergey's manual of systematic bacteriology, 2nd edn. Springer, New YorkGoogle Scholar
  18. Kappstein I, Grundmann H, Hauer T, Niemeyer C (2000) Aerators as a reservoir of Acinetobacter junii: an outbreak of bacteraemia in paediatric oncology patients. J Hosp Infect 44:27–30CrossRefGoogle Scholar
  19. Kim MH, Hao OJ, Wang NS (1997) Acinetobacter isolates from different activated sludge processes: characteristics and neural network identification. FEMS Microbiol Ecol 23:217–227CrossRefGoogle Scholar
  20. Kormas KA, Neofitou C, Pachiadaki M, Koufostathi E (2010) Changes of the bacterial assemblages throughout an urban drinking water distribution system. Environ Monit Assess 165:27–38CrossRefGoogle Scholar
  21. Krawczyk B, Lewandowski K, Kur J (2002) Comparative studies of the Acinetobacter genus and the species identification method based on the recA sequences. Mol Cell Probes 16:1–11CrossRefGoogle Scholar
  22. Kuo HY, Yang CM, Lin MF, Cheng WL, Tien N, Liou ML (2010) Distribution of blaOXA-carrying imipenem-resistant Acinetobacter spp. in 3 hospitals in Taiwan. Diagn Microbiol Infect Dis 66:195–199CrossRefGoogle Scholar
  23. La Scola B, Gundi VA, Khamis A, Raoult D (2006) Sequencing of the rpoB gene and flanking spacers for molecular identification of Acinetobacter species. J Clin Microbiol 44:827–832CrossRefGoogle Scholar
  24. Clinical and Laboratory Standards Institute (2007) Performance standards for antimicrobial susceptibility testing m100-s17. 27: 2Google Scholar
  25. Leclerc H, Moreau A (2002) Microbiological safety of natural mineral water. FEMS Microbiol Rev 26:207–222CrossRefGoogle Scholar
  26. Li D, Li Z, Yu J, Cao N, Liu R, Yang M (2010) Characterization of bacterial community structure in a drinking water distribution system during an occurrence of red water. Appl Environ Microbiol 76:7171–7180CrossRefGoogle Scholar
  27. Loret JF, Greub G (2010) Free-living amoebae: biological by-passes in water treatment. Int J Hyg Environ Health 213:167–175CrossRefGoogle Scholar
  28. Marshall C, Richards M, Black J, Sinickas V, Dendle C, Korman T, Spelman D (2007) A longitudinal study of Acinetobacter in three Australian hospitals. J Hosp Infect 67:245–252CrossRefGoogle Scholar
  29. McKeon DM, Calabrese JP, Bissonnette GK (1995) Antibiotic resistant gram-negative bacteria in rural groundwater supplies. Water Res 29:1902–1908CrossRefGoogle Scholar
  30. Medina J, Formento C, Pontet J, Curbelo A, Bazet C, Gerez J, Larranaga E (2007) Prospective study of risk factors for ventilator-associated pneumonia caused by Acinetobacter species. J Crit Care 22:18–26CrossRefGoogle Scholar
  31. Montealegre MC, Maya JJ, Correa A, Espinal P, Mojica MF, Ruiz SJ, Rosso F, Vila J, Quinn JP, Villegas MV (2012) First identification of OXA-72 carbapenemase from Acinetobacter pittii in Colombia. Antimicrob Agents Chemother. doi: 10.1128/AAC.05628-11
  32. Norton CD, LeChevallier MW (2000) A pilot study of bacteriological population changes through potable water treatment and distribution. Appl Environ Microbiol 66:268–276CrossRefGoogle Scholar
  33. Nowak A, Kur J (1995) Genomic species typing of acinetobacters by polymerase chain reaction amplification of the recA gene. FEMS Microbiol Lett 130:327–332CrossRefGoogle Scholar
  34. Pavlov D, de Wet CM, Grabow WO, Ehlers MM (2004) Potentially pathogenic features of heterotrophic plate count bacteria isolated from treated and untreated drinking water. Int J Food Microbiol 92:275–287CrossRefGoogle Scholar
  35. Pielou EC (1966) The measurement of diversity in different types of biological collections. J Theor Biol 13:131–144CrossRefGoogle Scholar
  36. Poirel L, Mansour W, Bouallegue O, Nordmann P (2008) Carbapenem-resistant Acinetobacter baumannii isolates from Tunisia producing the OXA-58-like carbapenem-hydrolyzing oxacillinase OXA-97. Antimicrob Agents Chemother 52:1613–1617CrossRefGoogle Scholar
  37. Poitelon JB, Joyeux M, Welte B, Duguet JP, Prestel E, Lespinet O, DuBow MS (2009) Assessment of phylogenetic diversity of bacterial microflora in drinking water using serial analysis of ribosomal sequence tags. Water Res 43:4197–4206CrossRefGoogle Scholar
  38. Quinteira S, Grosso F, Ramos H, Peixe L (2007) Molecular epidemiology of imipenem-resistant Acinetobacter haemolyticus and Acinetobacter baumannii isolates carrying plasmid-mediated OXA-40 from a Portuguese hospital. Antimicrob Agents Chemother 51:3465–3466CrossRefGoogle Scholar
  39. Regalado NG, Martin G, Antony SJ (2009) Acinetobacter lwoffii: bacteremia associated with acute gastroenteritis. Travel Med Infect Dis 7:316–317CrossRefGoogle Scholar
  40. Revetta RP, Pemberton A, Lamendella R, Iker B, Santo Domingo JW (2010) Identification of bacterial populations in drinking water using 16S rRNA-based sequence analyses. Water Res 44:1353–1360CrossRefGoogle Scholar
  41. Seifert H, Dijkshoorn L, Gerner-Smidt P, Pelzer N, Tjernberg I, Vaneechoutte M (1997) Distribution of Acinetobacter species on human skin: comparison of phenotypic and genotypic identification methods. J Clin Microbiol 35:2819–2825Google Scholar
  42. Shannon CE, Weaver W (1949) The mathematical theory of communication. University of Illinois Press, UrbanaGoogle Scholar
  43. Simões LC, Simões M, Vieira MJ (2010) Influence of the diversity of bacterial isolates from drinking water on resistance of biofilms to disinfection. Appl Environ Microbiol 76:6673–6679CrossRefGoogle Scholar
  44. Spence RP, Towner KJ, Henwood CJ, James D, Woodford N, Livermore DM (2002) Population structure and antibiotic resistance of Acinetobacter DNA group 2 and 13TU isolates from hospitals in the UK. J Med Microbiol 51:1107–1112Google Scholar
  45. Thangaraj K, Kapley A, Purohit HJ (2008) Characterization of diverse Acinetobacter isolates for utilization of multiple aromatic compounds. Bioresour Technol 99:2488–2494CrossRefGoogle Scholar
  46. Thomas V, Herrera-Rimann K, Blanc DS, Greub G (2006) Biodiversity of amoebae and amoeba-resisting bacteria in a hospital water network. Appl Environ Microbiol 72:2428–2438CrossRefGoogle Scholar
  47. Thomas V, Loret JF, Jousset M, Greub G (2008) Biodiversity of amoebae and amoebae-resisting bacteria in a drinking water treatment plant. Environ Microbiol 10:2728–2745CrossRefGoogle Scholar
  48. Thomas V, McDonnell G, Denyer SP, Maillard JY (2010) Free-living amoebae and their intracellular pathogenic microorganisms: risks for water quality. FEMS Microbiol Rev 34:231–259CrossRefGoogle Scholar
  49. Tognim MC, Andrade SS, Silbert S, Gales AC, Jones RN, Sader HS (2004) Resistance trends of Acinetobacter spp. In Latin America and characterization of international dissemination of multi-drug resistant strains: five-year report of the sentry antimicrobial surveillance program. Int J Infect Dis 8:284–291CrossRefGoogle Scholar
  50. Towner KJ (2009) Acinetobacter: An old friend, but a new enemy. J Hosp Infect 73:355–363CrossRefGoogle Scholar
  51. Van Looveren M, Goossens H, Steering Group ARPAC (2004) Antimicrobial resistance of Acinetobacter spp. in Europe. Clin Microbiol Infect 10:684–704CrossRefGoogle Scholar
  52. Vanbroekhoven K, Ryngaert A, Wattiau P, Mot R, Springael D (2004) Acinetobacter diversity in environmental samples assessed by 16S rRNA gene PCR-DGGE fingerprinting. FEMS Microbiol Ecol 50:37–50CrossRefGoogle Scholar
  53. Vaz-Moreira I, Egas C, Nunes OC, Manaia CM (2011a) Culture-dependent and culture-independent diversity surveys target different bacteria: a case study in a freshwater sample. Antonie Van Leeuwenhoek 100:245–257CrossRefGoogle Scholar
  54. Vaz-Moreira I, Novo A, Hantsis-Zacharov E, Lopes AR, Gomila M, Nunes OC, Manaia CM, Halpern M (2011b) Acinetobacter rudis sp. nov. isolated from raw milk and raw wastewater. Int J Syst Evol Microbiol 61:2837–2843CrossRefGoogle Scholar
  55. Vaz-Moreira I, Nunes OC, Manaia CM (2011c) Diversity and antibiotic resistance patterns of Sphingomonadaceae isolates from drinking water. Appl Environ Microbiol 77:5697–5706CrossRefGoogle Scholar
  56. Vaz-Moreira I, Nunes OC, Manaia CM (2012) Diversity and antibiotic resistance in Pseudomonas spp. from drinking water. Sci Total Environ 426:366–374CrossRefGoogle Scholar
  57. Villarreal JV, Schwartz T, Obst U (2010) Culture-independent techniques applied to food industry water surveillance—a case study. Int J Food Microbiol 141:S147–S155CrossRefGoogle Scholar
  58. Visca P, Seifert H, Towner KJ (2011) Acinetobacter infection—an emerging threat to human health. IUBMB Life 63:1048–1054CrossRefGoogle Scholar
  59. World Health Organisation (2008) Guidelines for drinking water. World Health Organization, GenevaGoogle Scholar
  60. Xi C, Zhang Y, Marrs CF, Ye W, Simon C, Foxman B, Nriagu J (2009) Prevalence of antibiotic resistance in drinking water treatment and distribution systems. Appl Environ Microbiol 75:5714–5718CrossRefGoogle Scholar
  61. Xu X, Kong F, Cheng X, Yan B, Du X, Gai J, Ai H, Shi L, Iredell J (2008) Integron gene cassettes in Acinetobacter spp. strains from south china. Int J Antimicrob Agents 32:441–445CrossRefGoogle Scholar
  62. Yamamoto S, Bouvet PJ, Harayama S (1999) Phylogenetic structures of the genus Acinetobacter based on gyrB sequences: comparison with the grouping by DNA-DNA hybridization. Int J Syst Bacteriol 49(Pt 1):87–95CrossRefGoogle Scholar
  63. Zhang Y, Marrs CF, Simon C, Xi C (2009) Wastewater treatment contributes to selective increase of antibiotic resistance among Acinetobacter spp. Sci Total Environ 407:3702–3706CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • Carlos Narciso-da-Rocha
    • 1
  • Ivone Vaz-Moreira
    • 1
    • 2
  • Liselott Svensson-Stadler
    • 3
  • Edward R. B. Moore
    • 3
  • Célia M. Manaia
    • 1
    Email author
  1. 1.CBQF, Escola Superior de BiotecnologiaUniversidade Católica PortuguesaPortoPortugal
  2. 2.LEPAE, Departamento de Engenharia Química, Faculdade de EngenhariaUniversidade do PortoPortoPortugal
  3. 3.CCUG, Culture Collection University of Gothenburg and Department of Infectious DiseaseSahlgrenska Academy of the University of GothenburgGothenburgSweden

Personalised recommendations