Antonie van Leeuwenhoek

, Volume 93, Issue 1–2, pp 111–122 | Cite as

Virulence and antibiotic susceptibility of Aeromonas spp. isolated from drinking water

  • Denise de Oliveira Scoaris
  • Jean Colacite
  • Celso V. Nakamura
  • Tânia Ueda-Nakamura
  • Benício A. de Abreu Filho
  • Benedito P. Dias Filho
Original Paper
First Online:
Received:
Accepted:

Abstract

Aeromonas isolates from tap water, mineral water, and artesian well water were investigated for their ability to produce different potential virulence factors or markers such as hemolysins, cytotoxins, phospholipase, DNase, hydrophobicity and their ability to adhere to epithelial cells and to abiotic surfaces. The susceptibility to antibiotics of Aeromonas isolates was also examined. Majority of the isolates displayed hemolytic activity against sheep erythrocytes, while only 7 of the 23 Aeromonas strains displayed DNase activity and 4 of the 23 Aeromonas strains tested were regarded as positive for phospholipase production. Most of the isolates showed cytotoxic activities in culture filtrate dilutions at titer of 1/8 or lower. No general relation between the strain isolated and the ability to interact with epithelial cells could be established. Using the bacterial adherence to hydrocarbons method, most of the strains were classified as highly hydrophilic. All five Aeromonas jandaei strains isolates, 9 of the 12 Aeromonas sp strains and four of the five Aeromonas hydrophila were multidrug resistant. The most active antimicrobial was ciprofloxacin (susceptible in 100% of the isolates), and the least active antibiotic was ampicillin (resistance in 92% of the isolates). The majority of the isolates tested were not killed by chlorine at 1.2 mg/l. Whether the high tolerance to chlorine of Aeromonas isolates can be linked to greater virulence is not know.

Keywords

Aeromonas Adherence Cytotoxicity Hydrophobicity Antimicrobial resistance 
This is a preview of subscription content, log in to check access.

Notes

Acknowledgments

This work was supported by the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Fundação Araucária, Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), and Programa de Pós-graduação em Microbiologia da Universidade Estadual de Londrina.

References

  1. American Public Health Association (19th) (1995) Standard methods for the examination of water and wastewater. APHA, WashingtonGoogle Scholar
  2. Anguita J, Rodríguez Aparício LB, Naharro G (1993) Purification, gene cloning, amino acid sequence analysis, and expression of an extracellular lipase from an Aeromonas hydrophila human isolate. Appl Environ Microbiol 59:2411–2417PubMedGoogle Scholar
  3. Baddour MD (1992) Extraintestinal Aeromonas infections. Mayo Clin Proc 67:496–498PubMedGoogle Scholar
  4. Baskin H, Dogan Y, Bahar IH, Yulug N (2002) Effect of subminimal inhibitory concentrations of three fluoroquinolones on adherence of uropathogenic strains of Escherichia coli. Int J Antimicrob Agents 19:79–82PubMedCrossRefGoogle Scholar
  5. Borrell N, Acinas SG, Figueras MJ, Martínez-Murcia AJ (1997) Identification of Aeromonas clinical isolates by restriction fragment length polymorphism of PCR-amplified 16S rRNA genes. J Clin Microbiol 35(7):1671–1674PubMedGoogle Scholar
  6. Bott TR, Miller PC (1983) Mechanisms of biofilm formation on aluminium tubes. J Chem Technol Biotechnol 3B:177–187Google Scholar
  7. Boussard P, Devleeschouwer MJ, Dony J (1989) Prevention of the adhesion of Pseudomonas aeruginosa to human buccal epithelial cells. Int J Pharmacol 53:253–256CrossRefGoogle Scholar
  8. Burke V, Cooper MJ, Robison J, Gracey M, Lesmana M, Echeverria P, Janda JM (1984b) Hemagglutination patterns of Aeromonas spp. in relation to biotype and source. J Clin Microbiol 19:39–43Google Scholar
  9. Burke V, Robinson J, Gracey M, Peterson D, Partridge K (1984a) Isolation of Aeromonas hydrophila grown a metropolitan water supply: seasonal correlation with clinical isolates. Appl Environ Microbiol 48:361–366PubMedGoogle Scholar
  10. Cahil M (1990) Virulence factors in motile Aeromonas species. J Appl Bacteriol 69:1–16Google Scholar
  11. Carrello A, Silburn KA, Budden JR, Chang BJ (1988) Adhesion of clinical and environmental Aeromonas isolates to HEp-2 cells. J Med Microbiol 26:19–27PubMedGoogle Scholar
  12. Cascon A, Yugueros J, Temprano A, Sanchez M, Hernanz C, Luengo JM, Naharro G (2000) A major secreted elastase is essential for pathogenicity of Aeromonas hydrophila. Infect Immun 68:3233–3241PubMedCrossRefGoogle Scholar
  13. Cerca N, Martins S, Sillankorva S, Jefferson KK, Gerald B, Píer GB, Rosario Oliveira R, Joana Azeredo J (2005), Effects of growth in the presence of subinhibitory concentrations of dicloxacillin on Staphylococcus epidermidis and Staphylococcus haemolyticus biofilms. Appl Environ Microbiol 71:8677–8682PubMedCrossRefGoogle Scholar
  14. Chamorey E, Fore M, Drancourt M (1999) An in-vitro evaluation of the activity of chlorine against environmental and nosocomial isolates of Aeromonas hydrophila. J Hosp Infect 41:45–49PubMedCrossRefGoogle Scholar
  15. Chang MC, Chang JC, Chen JP (1993) Cloning and nucleotidesequence of an extracellular a-amylase gene from Aeromonas hydrophila MCC-1. J Gen Microbiol 139:3215–3223PubMedGoogle Scholar
  16. Chang MC, Chang SY, Chen SL, Chuang SM (1992) Cloning and expression in Escherichia coli of the gene encoding an extracellular deoxyribonuclease (DNase) from Aeromonas hydrophila. Gene 122:175–180PubMedCrossRefGoogle Scholar
  17. Chen JP, Nagayama F, Chang MC (1991) Cloning and expression of a chitinase gene from Aeromonas hydrophila in Escherichia coli. Appl Environ Microbiol 57:2426–2428PubMedGoogle Scholar
  18. Chopra AK, Xu X-J, Ribardo D, Gonzalez M, Kuhl K, Petersen JW, Houston CW (2000) The cytotoxic enterotoxin of Aeromonas hydrophila induces proinflammatory cytokine production and activates arachidonic acid metabolism in macrophages. Infect Immun 68:2808–2818PubMedCrossRefGoogle Scholar
  19. Costerton JW, Cheng KJ, Geesy GG, Ladd TI, Nickel JC, Dasgupta M, Marie TJ (1987) Bacterial biofilm in nature and disease. Ann Rev Microbiol 41:435–464CrossRefGoogle Scholar
  20. Dal Sasso M, Culici M, Bovio C, Braga PC (2003) Gemifloxacin: effects of sub-inhibitory concentrations on various factors affecting bacterial virulence. Int J Antimicrob Agents 21:325–333PubMedCrossRefGoogle Scholar
  21. Diener M, Egleme C, Rummel W (1991) Phospholipase C-induced secretion and its interaction with carbachol in the rat colonic mucosa. Eur J Pharmacol 299:267–276CrossRefGoogle Scholar
  22. Edberg SC, Gallo P, Kontnick C (1996) Analysis of the virulence characteristics of bacteria isolated from bottled, water cooler, and tap water. Microb Ecol Health Dis 9:67–7712CrossRefGoogle Scholar
  23. Fainstein V, Weaver S, Bodey GP (1982) In vitro susceptibilities of Aeromonas hydrophila against new antibiotics. Antimicrob Agents Chemother 22:513–514PubMedGoogle Scholar
  24. Fiore AE, Michalski JM, Russell RG, Sears CL, Kaper JB (1997) Cloning, characterization, and chromosomal mapping of a phospholipase (lecithinase) produced by Vibrio cholerae. Infect Immun 65:3112–3117PubMedGoogle Scholar
  25. Focareta T, Manning PA (1991) Distinguishing between the extracellular DNases of Vibrio cholerae and development of a transformation system. Mol Microbiol 5:2547–2555PubMedCrossRefGoogle Scholar
  26. Francki KT, Chang BJ (1994) Variable expression of O-antigen and the role of lipopolysaccharide as an adhesin in Aeromonas sobria. FEMS Microbiol Lett 122:97–101PubMedCrossRefGoogle Scholar
  27. Gavín R, Merino S, Altarriba M, Canals R, Shaw JG, Toma´s JM (2003) Lateral flagella are required for increased cell adherence, invasion and biofilm formation by Aeromonas spp. FEMS Microbiol Lett 224:77–83PubMedCrossRefGoogle Scholar
  28. Gavín R, Rabaan AA, Merino S, Toma´s JM, Gryllos I (2002) Lateral flagella of Aeromonas species are essential for epithelial cell adherence and biofilm formation. Mol Microbiol 43:383–397PubMedCrossRefGoogle Scholar
  29. Ghanem EH, Missa ME, Eraki HM (1993) Aeromonas-associated gastroenteritis in Egypt. Zen bl Mikrobiol 148:441–447Google Scholar
  30. Hazenbos WL, Van den Berg BM, Von’t Wout JW, Mooi FR, Van Furth R (1994) Virulence factors determine attachment an ingestion of nonopsonized Bordetella pertusis by human monocytes. Infect Immun 62:4818–4824PubMedGoogle Scholar
  31. Herson DS, McGonigle B, Pouger MA, Baker KH (1987) Attachment as a factor in the protection of Enterobacter cloacae from chlorination. Appl Environ Microbiol 53:1178–1180PubMedGoogle Scholar
  32. Imzilm B, Lafdal YMO, Jana M (1996) Effect of wastewater stabilization ponds on antimicrobial susceptibility and haemolysin occurrence among motile Aeromonas strains. World J Microbiol Biotech 12:385–390CrossRefGoogle Scholar
  33. Ivanovska I (2003) Phospholipase as a factor of pathogenicity in microorganisms. J Mol Catal B 22:357–361CrossRefGoogle Scholar
  34. Janda JM (1991) Recent advances in the study of the taxonomy, pathogenicity and infectious syndromes associated with genus Aeromonas. Clin Microbiol Rev 4:397–410PubMedGoogle Scholar
  35. Janda JM, Bottone EJ (1981) Pseudomonas aeruginosa enzyme profiling: predictor of potential invasiveness and use as an epidemiological tool. J Clin Microbiol 14:55–60PubMedGoogle Scholar
  36. Johnson WM, Lior H (1981) Cytotoxicity and suckling mouse reactivity of Aeromonas hydrophila isolated from human sources. Can J Microbiol 27:1019–1027PubMedCrossRefGoogle Scholar
  37. King GE, Werner SB, Kizer KW (1992) Epidemiology of Aeromonas infections in California. Clin Infect Dis 15:449–452PubMedGoogle Scholar
  38. Kirov SM (2003) Bacteria that express lateral flagella enable dissection of the multifunctional roles of flagella in pathogenesis. FEMS Microbiol Lett 224:151–159PubMedCrossRefGoogle Scholar
  39. Kirov SM, Sanderson K, Dickson TC (1998) Characterisation of a type IV pilus produced by Aeromonas caviae. J Med Microbiol 47:527–531PubMedGoogle Scholar
  40. Koehler JM, Ashdown LR (1993) In vitro susceptibilities of tropical strains of Aeromonas species from Queensland, Australia, to 22 antimicrobial agents. Antimicrob Agents Chemother 37:905–907PubMedGoogle Scholar
  41. Krovacek K, Dumontet S, Eriksson E, Baloda B (1995) Isolation and virulence profiles of Aeromonas hydrophyla implicated in an outbreak of food poisoning in Sweden. Microbiol Immunol 39:655–661PubMedGoogle Scholar
  42. Kuhn I, Allestam G, Huys G, Janssen P, Kersters K, Krovacek K, Stenstrom T (1997) Diversity, persistence and virulence of Aeromonas strains isolated from drinking water distribution system in Sweden. Appl Environ Microbiol 63:2708–2715PubMedGoogle Scholar
  43. Le Chevallier MW, Evans TM, Seidler R, Daily OP, Merrel BR, Rollins DM, Joseph SW (1982) Aeromonas sobria in chlorinated drinking water supplies. Microbiol Ecol 8:325–333CrossRefGoogle Scholar
  44. Leake ES, Gristina AG, Wright MJ (1982) Use of chemotaxis chambers for studying in vitro bacterial colonization of biomaterials. J Clin Microbiol 15:320–323PubMedGoogle Scholar
  45. Ljungh A (1987) Aeromonas-toxins and other virulence factors. Experientia 43:367–368CrossRefGoogle Scholar
  46. McCarter L (1999) The multiple identities of Vibrio parahaemolyticus. J Mol Microbiol Biotechnol 1:51–57PubMedGoogle Scholar
  47. Merino S, Aguilar A, Nogueras MM, Regue M, Swift S, Tomas JM (1999) Cloning, sequencing, and role in virulence of two phospholipases (A1 and C) from mesophilic Aeromonas sp. serogroup O:34. Infect Immun 67:4008–4013PubMedGoogle Scholar
  48. Merino S, Rubires X, Aguilar A, Tomás JM (1996) The O:34-antigen lipopolysaccharide as an adhesin in Aeromonas hydrophila. FEMS Microbiol Lett 139:97–101PubMedGoogle Scholar
  49. Merino S, Rubires X, Knochel X, Tomás JM (1995) Emerging pathogens: Aeromonas spp. Int J Food Microbiol 28:157–168PubMedCrossRefGoogle Scholar
  50. Motyl MR, McKinley G, Janda JM (1985) In vitro susceptibilities of Aeromonas hydrophila, Aeromonas sobria, and Aeromonas caviae to 22 antimicrobial. Agents Antimicrob Agents Chemother 28:151–153Google Scholar
  51. Nacescu N, Israil A, Cedru C, Caplan D (1992) Hemolytic properties of some Aeromonas strains. Roum Arch Microbiol Immunol 51:147–156PubMedGoogle Scholar
  52. National Committee for Clinical Laboratory Standard (NCCLS) (2003) Performance standards for antimicrobial disk susceptibility tests. Approved standard, 8th ed. NCCLS document M2-A8. NCCLS, Wayne, Pa.Google Scholar
  53. National Committee for Clinical Laboratory Standard (NCCLS) (2004) Performance standards for antimicrobial disk susceptibility testing. Fourteenth informational supplement. NCCLS document M100-514. NCCLS, Wayne, Pa.Google Scholar
  54. Oliveira-Garcia D, Dall Agnol M, Rosales M, Azzuz ACGS, Alcântara N, Martinez MB, Giron JA (2003) Fimbriae and adherence of Stenotrophomonas maltophilia to epithelial cells and to abiotic surfaces. Cell Microbiol 5:625–636PubMedCrossRefGoogle Scholar
  55. Ørmen Ø, Østensvik Ø (2001) The occurrence of aerolysin-positive Aeromonas spp. and their cytotoxicity in Norwegian water sources. J Appl Microbiol 90:797–802PubMedCrossRefGoogle Scholar
  56. Polak A (1992) Virulence of Candida albicans mutants. Mycoses 35:9–16PubMedCrossRefGoogle Scholar
  57. Potomski J, Burke V, Watson I, Gracey M (1987) Purification of cytotoxic enterotoxin of Aeromonas sobria by use of monoclonal antibodies. J Med Microbiol 23:171–177PubMedGoogle Scholar
  58. Price MF, Wilkinson ID, Gentry LO (1982) Plate method for detection of phospholipase activity in Candida albicans. Sabouraudia 20:7–14PubMedGoogle Scholar
  59. Quinn DM, Atkinson HM, Bretag AH, Tester M, Trust TJ, Wong CYF, Flower RFLP (1994) Carbohydrate-reactive, pore-forming outer membrane proteins of Aeromonas hydrophila. Infect Immun 62:4054–4058PubMedGoogle Scholar
  60. Rabaan AA, Gryllos I, Toma´s JM, Shaw JG (2001) Motility and the polar flagellum are required for Aeromonas caviae adherence to HEp-2 cells. Infect Immun 69:4257–4267PubMedCrossRefGoogle Scholar
  61. Rivero O, Anguita J, Paniagua C, Naharro G (1990) Molecular cloning and characterization of an extracellular protease gene from Aeromonas hydrophila. J Bacteriol 172:3905–3908PubMedGoogle Scholar
  62. Rosenberg K, Gutnick D, Rosenberg E (1980) Adherence of bacteria to hydrocarbons: a simple method for measuring cell-surface hydrophobicity. FEMS Microbiol Lett 9:29–33CrossRefGoogle Scholar
  63. Schneider PF, Riley TV (1991) Cell surface hydrophobicity of Staphylococcus saprophyticus. Epidemiol Infect 106:71–75PubMedGoogle Scholar
  64. Seyfried PL, Frazer DJ (1980) Persistence of Pseudomonas aeruginosa in chlorinated swimming pools. Can J Microbiol 26:350–355PubMedCrossRefGoogle Scholar
  65. Shaw JF, Chang RC, Chuang KH, Yen YT, Wang FG (1994) Nucleotide sequence of novel arylesterase gene from Vibrio mimicus and characterization of the enzyme expressed in E. coli. Biochem J 298:675–680PubMedGoogle Scholar
  66. Shimada T, Sakazaki R, Horigome R, Uesaka Y, Niwano K (1984) Production of cholera-like enterotoxin by Aeromonas hydrophila. Jpn J Med Sci Biol 37:141–144PubMedGoogle Scholar
  67. Singh DV, Sanyal SC (1992) Enterotoxicity of clinical and environmental isolates of Aeromonas spp. J Med Microbiol 36(4):269–272PubMedGoogle Scholar
  68. Skehan P, Storeng R, Scudiero D, Monks A, McMahon J, Vistica D, Warren JT, Bokesch H, Kenney S, Boyd MR (1990) New colorimetric cytotoxicity assay for anti-cancer-drug screening. J Natl Cancer Inst 82:1107–1112PubMedCrossRefGoogle Scholar
  69. Snijder HJ, Dijkstra BW (2000) Bacterial phospholipase A: structure and function of an integral membrane phospholipase (BBA). Mol Cell Biol Lipids 1488:91–101CrossRefGoogle Scholar
  70. Takahashi A, Tanoue N, Nakano M, Hamamoto A, Okamoto K, Fujii Y, Harada N Nakaya Y (2005) A pore-forming toxin produced by Aeromonas sobria activates Ca2+ dependent Cl− secretion. Microb Pathog 38:173–180PubMedCrossRefGoogle Scholar
  71. Thornley JP, Shaw JG, Gryllos I, Eley A (1997) Virulence properties of clinically significant Aeromonas species. Rev Med Microbiol 8:61–72Google Scholar
  72. Thornley JP, Shaw JG, Gryllos IA, Eley A (1996) Adherence of Aeromonas caviae to human cell lines HEp-2 and Caco-2. J Med Microbiol 45:445–451PubMedCrossRefGoogle Scholar
  73. Thornton J, Howard SP, Buckley JT (1988) Molecular cloning of a phospholipid-cholesterol acyltransferase from Aeromonas hydrophila. Sequence homologies with lecithin-cholesterol acyltransferases and other lipases. Biochim Biophys Acta 959:153–159PubMedGoogle Scholar
  74. Titball RW (1993) Bacterial phospholipases C. Microbiol Rev 57:347–366PubMedGoogle Scholar
  75. Turnbull PC, Lee JV, Miliotis MD, Van de Walle S, Koornhof HJ, Jeffery L, Bryant TN (1984) Enterotoxin production in relation to taxonomic grouping and source of isolation of Aeromonas species. J Clin Microbiol 19:175–180PubMedGoogle Scholar
  76. Vranes J (1996) Effect of subinhibitory concentrations of ceftazidime, ciprofloxacin and azithromycin on the hemagglutination and adherence of uropathogenic Escherichia coli strains. Chemotherapy 42:177–185PubMedCrossRefGoogle Scholar
  77. Walsh TR, Payne DJ, MacGowan AP, Bennett PM (1995) A clinical isolate of Aeromonas sobria with three chromosomally mediated inducible beta-lactamases: a cephalosprinase, a penicillinase and a third enzyme, displaying carbapenemase activity. J Antimicrob Chemother 35:271–279PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2007

Authors and Affiliations

  • Denise de Oliveira Scoaris
    • 1
  • Jean Colacite
    • 2
  • Celso V. Nakamura
    • 3
  • Tânia Ueda-Nakamura
    • 3
  • Benício A. de Abreu Filho
    • 3
  • Benedito P. Dias Filho
    • 3
  1. 1.Programa de Pós-graduação em MicrobiologiaUniversidade Estadual de LondrinaLondrinaBrazil
  2. 2.Programa de Pós-graduação em Análises ClínicasUniversidade Estadual de MaringáMaringáBrazil
  3. 3.Departamento de Análises ClínicasUniversidade Estadual de MaringáMaringáBrazil

Personalised recommendations