Skip to main content
SpringerLink
Log in

No effect of N-acyl homoserine lactones disruption by lactonase enzyme on the virulence of Vibrio anguillarum towards sea bass (Dicentrarchus labrax) and brine shrimp (Artemia franciscana)

  • Published:
Aquaculture International Aims and scope Submit manuscript

Abstract

Quorum sensing (QS) is a bacterial communication system in which bacteria coordinate the expression of certain genes in response to the presence of small signal molecules. Some signal molecules called autoinducers are found to regulate several phenotypes of bacteria. The molecules used for the QS system of Vibrio anguillarum are supposed to regulate their pathogenic genes. However, it is not well understood how exactly the virulence gene of V. anguillarum is regulated by the QS molecules to date. To address this question, one of the QS molecules of the NB10 strain of V. anguillarum called N-acyl homoserine lactones (AHLs) was disrupted by a lactonase enzyme to verify its role on their virulence. The effects of AHLs degradation in vitro were investigated by measuring its effect on growth and virulence factors production (lipase, phospholipase, and caseinase) and in vivo by verifying its effect on sea bass and Artemia larvae infection. Lactonase enzyme degraded AHLs of NB10; yet that degradation had no effect on the cell growth and production of the tested virulence factors. Similarly, challenge tests demonstrated that larval mortality was not significantly lower by lactonase treatment. Therefore, these observations indicated that virulence gene expression of V. anguillarum and its virulence are not regulated by the AHL-mediated QS system; either different molecules or other factors are associated with the virulence regulation of V. anguillarum towards the tested life stages of sea bass and Artemia.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  • Akinbowale OL, Peng H, Barton MD (2007) Diversity of tetracycline resistance genes in bacteria from aquaculture sources in Australia. J Appl Microbiol 103:2016–2025

    Article  CAS  PubMed  Google Scholar 

  • Abuaita BH, Withey JH (2009) Bicarbonate induces Vibrio cholerae virulence gene expression by enhancing ToxT activity. Infect Immun 77:4111–4120

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Allison D, Ruiz B, SanJose C, Jaspe A, Gilbert P (1998) Extracellular products as mediators of the formation and detachment of Pseudomonas fluorescens biofilms. FEMS Microbiol Lett 167:179–184

    Article  CAS  PubMed  Google Scholar 

  • Beck von Bodman S, Farrand SK (1995) Capsular polysaccharide biosynthesis and pathogenicity in Erwinia stewartii require induction by an N-acyl homoserine lactone autoinducer. J Bacteriol 177:5000–5008

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Buchholtz C, Nielsen KF, Milton DL, Larsen JL, Gram L (2006) Profiling of acylated homoserine lactones of Vibrio anguillarum in vitro and in vivo: influence of growth conditions and serotype. Syst Appl Microbiol 29:433–445

    Article  CAS  PubMed  Google Scholar 

  • Bruhn JB, Dalsgaard I, Nielsen KF, Buchholtz C et al (2005) Quorum sensing signal molecules (acylated homoserine lactones) in Gram-negative fish pathogenic bacteria. Dis Aquat Org 65:43–52

    Article  CAS  PubMed  Google Scholar 

  • Buch C, Sigh J, Nielsen J, Larsen JL, Gram L (2003) Production of acylated homoserine lactones by different serotypes of Vibrio anguillarum both in culture and during infection of rainbow trout. Syst Appl Microbiol 26:338–349

    Article  CAS  PubMed  Google Scholar 

  • Costa JM, Loper JE (1997) EcbI and EcbR: homologs of LuxI and LuxR affecting antibiotic and exoenzyme production by Erwinia carotovora sub sp. Beta vasculorum. Can J Microbiol 43:1164–1171

    Article  CAS  PubMed  Google Scholar 

  • Croxatto A, Lauritz J, Chen C, Milton DL (2007) Vibrio anguillarum colonization of rainbow trout integument requires a DNA locus involved in exopolysaccharide transport and biosynthesis. Environ Microbiol 9:370–382

    Article  CAS  PubMed  Google Scholar 

  • Defoirdt T, Bossier P, Sorgeloos P, Verstraete W (2005) The impact of mutations in the quorum sensing systems of Aeromonas hydrophila, Vibrio anguillarum and Vibrio harveyi on their virulence towards gnotobiotically cultured Artemia franciscana. Environ Microbiol 7:1239–1247

    Article  CAS  PubMed  Google Scholar 

  • Dierckens K, Rekecki A, Laureau S, Sorgeloos P et al (2009) Development of a bacterial challenge test for gnotobiotic sea bass (Dicentrarchus labrax) larvae. Environ Microbiol 11:526–533

    Article  CAS  PubMed  Google Scholar 

  • Donabedian H (2003) Quorum sensing and its relevance to infectious disease. J Inf Secur 46:207–214

    CAS  Google Scholar 

  • Dong YH, JL X, Li XZ, Zhang LH (2000) AiiA, an enzyme that inactivates the acylhomoserine lactone quorum-sensing signal and attenuates the virulence of Erwinia carotovora. P Natl Acad Sci USA 97:3526–3531

    Article  CAS  Google Scholar 

  • Dong YH, Wang LH, JL X, Zhang HB et al (2001) Quenching quorum-sensing-dependent bacterial infection by an N-acyl homoserine lactonase. Nature 411:813–817

    Article  CAS  PubMed  Google Scholar 

  • Dong YH, Gusti AR, Zhang Q, JL X, Zhang LH (2002) Identification of quorum-quenching N-acyl homoserine lactonases from Bacillus species. Appl Environ Microbiol 68:1754–1759

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Dong YH, Zhang XF, JL X, Zhang LH (2004) Insecticidal Bacillus thuringiensis silences Erwinia carotovora virulence by a new form of microbial antagonism, signal interference. Appl Environ Microbiol 70:954–960

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Eberl L, Winson MK, Sternberg C, Stewart GSAB et al (1996) Involvement of N-acyl-L-homoserine lactone autoinducers in controlling the multicellular behaviour of Serratia liquefaciens. Mol Microbiol 20:127–136

    Article  CAS  PubMed  Google Scholar 

  • Gilk SD, Cockrell DC, Luterbach C, Hansen B et al (2013) Bacterial colonization of host cells in the absence of cholesterol. PLoS Pathog 9

  • Gotoh K, Kodama T, Hiyoshi H, Izutsu K et al (2010) Bile acid induced virulence gene expression of Vibrio parahaemolyticus reveals a novel therapeutic potential for bile acid sequestrants. PLoS One 5(10):e13365

    Article  PubMed  PubMed Central  Google Scholar 

  • Hsiao A, Liu Z, Joelsson A, Zhu J (2006) Vibrio cholerae virulence regulator-coordinated evasion of host immunity. P Natl Acad Sci USA 103:14542–14547

    Article  CAS  Google Scholar 

  • LaSarre B, Federle MJ (2013) Exploiting quorum sensing to confuse bacterial pathogens. Microbiol Mol Biol Rev 77(1):73–111

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lee SJ, Park SY, Lee JJ, Yum DY et al (2002) Genes encoding the N-acyl homoserine lactone-degrading enzyme are widespread in many subspecies of Bacillus thuringiensis. Appl Environ Microbiol 68:3919–3924

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Li X, Yang Q, Dierckens K, Milton DL, Defoirdt T (2014) RpoS and indole signaling control the virulence of Vibrio anguillarum towards gnotobiotic sea bass (Dicentrarchus labrax) larvae. PLoS One 9(10):e111801

    Article  PubMed  PubMed Central  Google Scholar 

  • Lin YH, JL X, Hu J, Wang LH et al (2003) Acyl-homoserine lactone acylase from Ralstonia sp. strain XJ12B represents a novel and potent class of quorum-quenching enzymes. Mol Microbiol 47:849–860

    Article  PubMed  Google Scholar 

  • Ma L, Chen J, Liu R, Zhang XH, Jiang YA (2009) Mutation of rpoS gene decreased resistance to environmental stresses, synthesis of extracellular products and virulence of Vibrio anguillarum. FEMS Microbiol Ecol 70:130–136

    Article  PubMed  Google Scholar 

  • McGee K, Horstedt P, Milton DL (1996) Identification and characterization of additional flagellin genes from Vibrio anguillarum. J Bacteriol 178:5188–5198

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Milton DL (2006) Quorum sensing in vibrios: complexity for diversification. Int J Med Microbiol 296:61–71

    Article  CAS  PubMed  Google Scholar 

  • Molina L, Constantinescu F, Michel L, Reimmann C et al (2003) Degradation of pathogen quorum-sensing molecules by soil bacteria: a preventive and curative biological control mechanism. FEMS Microbiol Ecol 45:71–81

    Article  CAS  PubMed  Google Scholar 

  • Natrah FMI, Ruwandeepika HAD, Pawar S, Karunasagar I et al (2011) Regulation of virulence factors by quorum sensing in Vibrio harveyi. Vet Microbiol 154:124–129

    Article  CAS  PubMed  Google Scholar 

  • Ni N, Li M, Wang J, Wang B (2009) Inhibitors of bacterial quorum sensing. Med Res Rev 29:65–124

    Article  CAS  PubMed  Google Scholar 

  • Norqvist A, Norrman B, Wolf-Watz H (1990) Identification and characterization of a zinc metallo protease associated with invasion by the fish pathogen Vibrio anguillarum. Infect Immun 58:3731–3736

    CAS  PubMed  PubMed Central  Google Scholar 

  • Paillard C, Leroux F, Borrego JJ (2004) Bacterial disease in marine bivalves: review of recent studies: trends and evolution. Aquat Living Resour 17:477–498

    Article  Google Scholar 

  • Phillips I, Casewell M, Cox T, Groot BD et al (2004) Does the use of antibiotics in food animals pose a risk to human health? A critical review of published data. J Antimicrob Chemother 53(1):28–52

    Article  CAS  PubMed  Google Scholar 

  • Rasch M, Buch C, Austin B, Slierendrecht WJ et al (2004) An inhibitor of bacterial quorum sensing reduces mortalities caused by vibriosis in rainbow trout (Oncorhynchus mykiss, Walbaum). Syst Appl Microbiol 27:350–359

    Article  CAS  PubMed  Google Scholar 

  • Rock JL, Nelson DR (2006) Identification and characterization of a hemolysin gene cluster in Vibrio anguillarum. Infect Immun 74:2777–2786

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Smith VJ, Brown JH, Hauton C (2003) Immunostimulation in crustaceans: does it really protect against infection? Fish Shellfish Immunol 15:71–90

    Article  PubMed  Google Scholar 

  • Stork M, Di Lorenzo M, Welch TJ, Crosa JH (2007) Transcription termination within the iron transport-biosynthesis operon of Vibrio anguillarum requires an antisense RNA. J Bacteriol 189:3479–3488

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Subasinghe RP, Bondad-Reantaso MG, McGladdery SE (2001) Aquaculture development, health and wealth. In: Subasinghe RP, Bueno P, Phillips MJ, Hough C et al (eds) Aquaculture in the Third Millennium, Bangkok, 20–25 February 2000

  • Teplitski M, Robinson JB, Bauer WD (2000) Plants secrete substances that mimic bacterial N-acyl homoserine lactone signal activities and affect population density-dependent behaviors in associated bacteria. Mol Plant-Microbe Interact 13:637–648

    Article  CAS  PubMed  Google Scholar 

  • Tinh NTN, Dung NV, Trung CT, Thuy VT (2013) In vitro characterization of a recombinant AHL-lactonase from Bacillus cereus isolated from a striped catfish (Pangasianodon hypophthalmus) pond. Indian J Microbiol 53(4):485–487

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Tinh NTN, Dung NV, Thuy VT, Suong NT (2014) Inhibition of quorum sensing-mediated bioluminescence in Vibrio harveyi by a recombinant AHL-lactonase from Bacillus cereus. J Pure Appl Microbiol 8:353–360

    Google Scholar 

  • Toranzo AE, Magarinos B, Romalde JL (2005) A review of the main bacterial fish diseases in mariculture systems. Aquaculture 246:37–61

    Article  Google Scholar 

  • Xu F, Byun T, Dussen HJ, Duke KR (2003) Degradation of N-acyl homoserine lactones the bacterial quorum-sensing molecules by acylase. J Biotechnol 101:89–96

    Article  CAS  PubMed  Google Scholar 

  • Yang H, Chen J, Yang G, Zhang XH, Li Y (2007) Mutational analysis of the zinc metallo protease EmpA of Vibrio anguillarum. FEMS Microbiol Lett 267:56–63

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This study was carried out under the Scientific Research Fund of Flanders (FWO-Vlaanderen project no 1.5.013.12N) and the Special Research Fund of Ghent University (GOA project no BOF12/GOA/022). In addition, a part of this study was supported by the Vietnamese National Foundation for Science and Technology Development (NAFOSTED), agreement No. 05/2010/HÐ-KHTN. We are also grateful to Artemia Reference Center, Ghent University, Belgium and Research Institute for Aquaculture No. 2, Hochiminh City, Vietnam, for providing technical and logistic supports.

Author information

Authors and Affiliations

  1. Laboratory of Aquaculture & Artemia Reference Center, Ghent University, Coupure Links 653, B-9000, Gent, Belgium

    Mousumi Das, Xuan Li & Peter Bossier

  2. Bangabandhu Sheikh Mujibur Rahman Agricultural University, Gazipur, Bangladesh

    Mousumi Das

  3. Research Institute for Aquaculture No. 2, Hochiminh City, Vietnam

    Nguyen Viet Dung & Nguyen Thi Ngoc Tinh

Authors
  1. Mousumi Das
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Xuan Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Nguyen Viet Dung
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Peter Bossier
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Nguyen Thi Ngoc Tinh
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Peter Bossier.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Das, M., Li, X., Dung, N.V. et al. No effect of N-acyl homoserine lactones disruption by lactonase enzyme on the virulence of Vibrio anguillarum towards sea bass (Dicentrarchus labrax) and brine shrimp (Artemia franciscana). Aquacult Int 26, 363–374 (2018). https://doi.org/10.1007/s10499-017-0214-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10499-017-0214-2

Keywords

Search

Navigation