Stenotrophomonas maltophilia AHL-Degrading Strains Isolated from Marine Invertebrate Microbiota Attenuate the Virulence of Pectobacterium carotovorum and Vibrio coralliilyticus
Many Gram-negative aquacultural and agricultural pathogens control virulence factor expression through a quorum-sensing (QS) mechanism involving the production of N-acylhomoserine (AHL) signalling molecules. Thus, the interruption of QS systems by the enzymatic degradation of signalling molecules, known as quorum quenching (QQ), has been proposed as a novel strategy to combat these infections. Given that the symbiotic bacteria of marine invertebrates are considered to be an important source of new bioactive molecules, this study explores the presence of AHL-degrading bacteria among 827 strains previously isolated from the microbiota of anemones and holothurians. Four of these strains (M3-1, M1-14, M3-13 and M9-54-2), belonging to the species Stenotrophomonas maltophilia, were selected on the basis of their ability to degrade a broad range of AHLs, and the enzymes involved in their activity were identified. Strain M9-54-2, which showed the strongest AHL-degrading activity, was selected for further study. High-performance liquid chromatography–mass-spectrometry confirmed that the QQ enzyme is not a lactonase. Strain M9-54-2 degraded AHL accumulation and reduced the production of enzymatic activity in Pectobacterium carotovorum CECT 225T and Vibrio coralliilyticus VibC-Oc-193 in in vitro co-cultivation experiments. The effect of AHL inactivation was confirmed by a reduction in potato tuber maceration and brine shrimp (Artemia salina) mortality caused by P. carotovorum and Vibrio coralliilyticus, respectively. This study strengthens the evidence of marine organisms as an underexplored and promising source of QQ enzymes, useful to prevent infections in aquaculture and agriculture. To our knowledge, this is the first time that anemones and holothurians have been studied for this purpose.
KeywordsQuorum quenching N-Acylhomoserine lactones Stenotrophomonas Acylase
José Carlos Reina is supported by an FPU fellowship from the Spanish Ministry of Education, Culture and Sport (FPU15/01717). The authors wish to thank Michael O’Shea for proofreading the manuscript.
This study was funded by the Spanish Ministry of the Economy and Competitiveness [AGL2015-68806-R].
Compliance with Ethical Standards
Conflicts of Interest
The authors declare that they have no conflict of interest.
- Chan KG, Atkinson S, Mathee K, Sam CK, Chhabra SR, Cmara M, Koh CL, Williams P (2011) Characterization of N-acylhomoserine lactone-degrading bacteria associated with the Zingiber officinale (ginger) rhizosphere: co-existence of quorum quenching and quorum sensing in Acinetobacter and Burkholderia. BMC Microbiol 11:51CrossRefGoogle Scholar
- Crépin A, Barbey C, Beury-Cirou A, Hélias V, Taupin L, Reverchon S, Nasser W, Faure D, Dufour A, Orange N, Feuilloley M, Heurlier K, Burini JF, Latour X (2012) Quorum sensing signaling molecules produced by reference and emerging soft-rot bacteria (Dickeya and Pectobacterium spp.). PLoS One 7:e35176CrossRefGoogle Scholar
- Dong YH, Zhang LH (2005) Quorum sensing and quorum-quenching enzymes. J Microbiol 43:101–109Google Scholar
- Food and Agriculture Organization (FAO) (2016) The state of world fisheries and aquaculture 2016. Food and Agriculture Organization of the United Nations, Rome, ItalyGoogle Scholar
- Furushita M, Okamoto A, Maeda T, Ohta M, Shiba T (2005) Isolation of multidrug-resistantStenotrophomonas maltophilia from cultured yellowtail (Seriola quinqueradiata) from a marine fish farm. Appl Environ Microbiol 71:5598–5600Google Scholar
- Plate-based assay for swimming motility in Pseudomonas aeruginosa. Methods Mol Biol 1149:59–65Google Scholar
- Ha D-G, Kuchma SL, O'Toole GA (2014c) Plate-based assay for swarming motility in Pseudomonas aeruginosa. Methods Mol Biol 1149:67–72Google Scholar
- Hughes C, Fenical W (2012) Antibacterials from the sea. Chemistry (Easton) 42:12512–12525Google Scholar
- Jeffries CD, Holtmian DF, Guse DG (1957) Rapid method for determining the activity of microorganisms on nucleic acids. J Bacteriol 73:590–591Google Scholar
- Marchler-Bauer A, Lu S, Anderson JB, Chitsaz F, Derbyshire MK, DeWeese-Scott C, Fong JH, Geer LY, Geer RC, Gonzales NR, Gwadz M, Hurwitz DI, Jackson JD, Ke Z, Lanczycki CJ, Lu F, Marchler GH, Mullokandov M, Omelchenko MV, Robertson CL, Song JS, Thanki N, Yamashita RA, Zhang D, Zhang N, Zheng C, Bryant SH (2011) CDD: a conserved domain database for the functional annotation of proteins. Nucleic Acids Res 39:D225–D229CrossRefGoogle Scholar
- Marchler-Bauer A, Derbyshire MK, Gonzales NR, Lu S, Chitsaz F, Geer LY, Geer RC, He J, Gwadz M, Hurwitz DI, Lanczycki CJ, Lu F, Marchler GH, Song JS, Thanki N, Wang Z, Yamashita RA, Zhang D, Zheng C, Bryant SH (2015) CDD: NCBI’s conserved domain database. Nucleic Acids Res 43:D222–D226CrossRefGoogle Scholar
- Marchler-Bauer A, Bo Y, Han L, He J, Lanczycki CJ, Lu S, Chitsaz F, Derbyshire MK, Geer RC, Gonzales NR, Gwadz M, Hurwitz DI, Lu F, Marchler GH, Song JS, Thanki N, Wang Z, Yamashita RA, Zhang D, Zheng C, Geer LY, Bryant SH (2017) CDD/SPARCLE: functional classification of proteins via subfamily domain architectures. Nucleic Acids Res 45:D200–D203CrossRefGoogle Scholar
- Mcclean KH, Winson MK, Fish L, Taylor A, Chhabra SR, Camara M, Daykin M, John H, Swift S, Bycroft BW, Stewart GS, Williams P (1997) Quorum sensing and Chromobacterium violaceum: exploitation of violacein production and inhibition for the detection of N-acyl homoserine lactones. Microbiology 143:3703–3711CrossRefGoogle Scholar
- Mukherjee P, Roy P (2016) Genomic potential of Stenotrophomonas maltophilia in bioremediation with an assessment of its multifaceted role in our environment. Front Microbiol 7:1–14Google Scholar
- Pikovskaya RI (1948) Mobilization of phosphorus in soil in connection with the vital activity of some microbial species. Mikrobiologiya 17:362–370Google Scholar
- Quiñones B, Dulla G, Lindow SE (2005) Exopolysaccharide production, motility, and virulence in Pseudomonas syringae. Society 18:682–693Google Scholar
- Torres M, Reina JC, Fuentes-Monteverde JC, Fernandez G, Rodriguez J, Jiménez C, Llamas I (2018) AHL-lactonase expression in three marine emerging pathogenic Vibrio spp. reduces virulence and mortality in brine shrimp (Artemia salina) and Manila clam (Venerupis philippinarum). PLoS One 13:1–23Google Scholar
- Yates EA, Philipp B, Buckley C, Atkinson S, Chhabra SR, Sockett RE, Goldner M, Dessaux Y, Cámara M, Smith H, Williams P (2002) N-acylhomoserine lactones undergo lactonolysis in a pH-, temperature-, and acyl chain length-dependent manner during growth of Yersinia pseudotuberculosis and Pseudomonas aeruginosa. Infect Immun 70:5635–5646CrossRefGoogle Scholar
- Zaman SB, Hussain MA, Nye R, Mehta V, Mamun KT, Hossain N (2017) A review on antibiotic resistance: alarm bells are ringing. Cureus 9:e1403Google Scholar