Vibrionaceae

Vibrionaceae nigripulchritudo by Le Roux
Description: 

Bacteria from the genus Vibrio are dominant culturable species of the marine ecosystems. Some are human human pathogens (for instance V. cholerae) but the vast majority are species responsible for marine animal diseases.

These bacteria can colonize numerous habitats, thrive as free, planktonic living organisms, live in biofilms, or form various associations with their hosts, from symbiosis to commensalism and pathogeny. This adaptability is linked to their capacity to generate genomic diversity (i.e. genomic plasticity), which implies material genetic acquisition through tranformation (by intake of naked DNA), conjugation (a plasmid is transfered from a donor to a recipient cell), or transduction (a bacteriophage injects its DNA into a bacterial cell). In certain cases, exogenous DNA can establich itself as an independent replicon (plasmid, episome), in other cases, it is integrated into the genome of the recipient (transposon, integron, integrative conjugative element). Acquisition of new genetic resources can modify the bacterium genotype, confering a selective advantage (antibiotic resistance, virulence).

Vibrio genome comprises two chromosomes. Chromosome I encodes genes involved in essential cell functions, whereas Chromosome II would be important for genomic variability and hence phenotypic variations. Hence, in Vibrio, evolution of virulence is tightly linked to the genome structural feature.

Vibrionaceae encompasses 141 species (http://www.bacterio.cict.fr/) distributed into five genera: Vibrio (109), Photobacterium (23), Salinivibrio (4), Enterovibrio (4) and Grimontia (1) (Sawabe et al., 200710)(Thompson et al., 200914)(Thompson et al., 20046). Currently, 320 genomes (22 completed) are available on Genbank.

Genetic resources: 

Available individuals

  • V. nigripulchritudo : 20 mutants (Δ chromosomic genes, transcomplementation of the nigritoxin, plasmids curing)
  • V. coralliilyticus : 3 mutants (Δ metalloprotease genes)
  • V. splendidus : 49 mutants (Δ metalloprotease genes, Δ quorum sensing genes, Δ other putative virulence genes...)
  • V. parahaemolyticus : 2 mutants (hemolysin, component of the system of type III secretion system)
  • V. tasmaniensis : cloning of toxins and metalloproteases

Work in progress :

  • P. angustum and differents strains of group V. splendidus

Toolbox

PLASMIDS :

 

E.coli FOR STRAINS MANIPULATION :

 

Culture: 

The bacterial strains and plasmids used in this study are described in Tables X and Y. V. splendidus strain LGP32 was isolated from the hemolymph of oysters suffering from outbreaks of summer mortalities, and demonstrated to be pathogenic for oysters and clams (23). V. cholerae and E. coli strains were grown in Luria-Bertani (LB) or, in case of P3813, Mueller-Hinton (MH) broth, at 37°C. V. splendidus strains were grown in LB-NaCl 0.5M, marine broth (MB) or marine agar (MA) at 20°C. All media were from Difco. Antibiotics were used at the following concentrations : ampicillin (Ap), 100 ug/ml ; chloramphenicol (Cm), 12,5ug/ml ; erythromycin (Erm), 200 ug/ml ; kanamycin (Km), 25 ug/ml ; nalidixic acid (Nal), 30 ug/ml ; spectinomycin (Sp), 50 ug/ml ; tetracycline (Tc), 15 ug/ml. Thymidine (dT) and diaminopimelate (DAP) were supplemented when necessary to a final concentration of 0.3mM. Induction of ccdB expression under the control of PBAD promoter was achieved by the addition of 0,2% L-arabinose to the growth media and repressed by 1% D-glucose.

Contact: 

Frederique Le Roux - Head of the GV team Equipe émergente Ifremer/UPMC: "Génomique des Vibrio"- Station Biologique, Place Georges Teissier,  CS 90074, 29688 Roscoff, France
 

References: 
  1. Le Roux F., Gay M., Lambert C., Waechter M., Poubalanne S., Chollet B., Nicolas J.L., Berthe F.C. (2002) Comparative analysis of Vibrio splendidus related strains isolated during Crassostrea gigas mortality events. Aquatic Living Resources, 15 : 251-258.
  2. Gay M., Berthe F.C., Le Roux F. (2004). Screening of Vibrio isolates towards the development of an experimental infection model in the Pacific oyster, Crassostrea gigas. Disease of Aquatic Organisms, 59 : 49-56.
  3. Gay M., Renault T., Pons A. M., Le Roux F. (2004). Two Vibrio splendidus-related strains collaborate to kill Crassostrea gigas, taxonomy and host alterations. Disease of Aquatic Organisms. 62 :1-16.
  4. Faury N., Saulnier D., Thompson F. L., Gay M., Swings J., Le Roux F. (2004). Vibrio crassostreae sp.nov., isolated from the hemolymph of oysters (Crassostrea gigas). International Journal of Systematic and Evolutionnary Microbiology. 54 : 2137-2140.
  5. Le Roux F., Gay M., Lambert C., Nicolas J.L., Gouy M., Berthe F.C. (2004). Phylogenetic study and identification of Vibrio splendidus related strains based on gyrB gene sequences. Disease of Aquatic Organisms, 58 : 143-150.
  6. Thompson FL, Iida T, Swings J: Biodiversity of vibrios. Microbiol Mol Biol Rev 2004, 68(3):403-431.
  7. Le Roux F., Goubet A., Thompson F. L., Faury N., Gay M., Swings J., Saulnier D. (2005) Vibrio gigantis sp. nov., isolated from the haemolymph of cultured oysters (Crassostrea gigas) International Journal of Systematic and Evolutionnary Microbiology 55 :2251-2255.
  8. Goarant C, Y Reynaud, D Ansquer, S de Decker, D Saulnier & F le Roux. (2006) Molecular epidemiology of Vibrio nigripulchritudo, a pathogen of cultured penaeid shrimp (Litopenaeus stylirostris) in New Caledonia Systemic and Applied Microbiology, 29(7) :570-80.
  9. Le Roux F, Binesse J, Saulnier D, Mazel D. (2007). Construction of a Vibrio splendidus mutant lacking the metalloprotease gene vsm by use of a novel counterselectable suicide vector. Appl Environ Microbiol. 73(3):777-84.
  10. Sawabe T., Kita-Tsukamoto K. & Thompson F. L. (2007). Inferring the evolutionary history of vibrios by means of multilocus sequence analysis. J Bacteriol. 189, 7932–7936.
  11. Binesse J. , Delsert C., Champomier-Vergès M-C., Saulnier D., Zagorec M., Mazel D. and Le Roux F. (2008) The Metalloprotease Vsm is the Main Toxic Factor for Vibrio splendidus Secretome ; Appl Environ Microbiol. 74 :7108-7117.
  12. Reynaud Y., Saulnier D., Mazel D., Goarant C. and Le Roux F. (2008) Identification of a plasmid associated with virulence in Vibrio nigripulchritudo, a pathogen of the shrimp Litopenaeus stylirostris ; Appl Environ Microbiol 74 :3038-3047.
  13. Le Roux F., Zouine M., Chakroun N., Binesse J., Saulnier D., Goarant C Bouchier C., Zidane N., Ma L., Rusniok C., Buchrieser C., Polz MF., Mazel D. Genome sequence of Vibrio splendidus: a dominant group of bacterioplankton presenting a large genotypic diversity (2009). Environ Microbiol. 11(8):1959-70.
  14. Thompson C.C., Vicente A.C., Souza R.C., Vasconcelos A.T., Vesth T., Alves N. Jr., Ussery D.W., Lida T., Thompson F.L. (2009) Genomic taxonomy of vibrios. BMC Evol Biol. 9:258.
  15. Duperthuy M, Binesse J, Le Roux F, Romestand B, Caro A, Got P, Givaudan A, Mazel D, Bachère E, Destoumieux-Garzón D. (2010) The major outer membrane protein OmpU of Vibrio splendidus contributes to host antimicrobial peptide resistance and is required for virulence in the oyster Crassostrea gigas. Environ Microbiol. 12(4):951-63.
  16. Labreuche Y, Le Roux F, Henry J, Zatylny C, Huvet A, Lambert C, Soudant P, Mazel D, Nicolas JL. (2010) Vibrio aestuarianus zinc metalloprotease causes lethality in the Pacific oyster Crassostrea gigas and impairs the host cellular immune defenses. Fish Shellfish Immunol. 29(5):753-8.
  17. Duperthuy M, Schmitt P, Garzónd E, Caro A, Rosab R, Le Roux F, Lautrédou-Audouy N, Got P, Romestand B, de Lorgeril J, Kieffer-Jaquinod S, Bachère E, Destoumieux-Garzón D. (2011) Use of OmpU porins for attachment and invasion of Crassostrea gigas immune cells by the oyster pathogen Vibrio splendidus. Proc Natl Acad Sci U S A.108 (7): 2993-8.
  18. Le Roux F, Labreuche Y., Davis B.M., Iqbal N., Mangenot S., Goarant C., Mazel D., Waldor M.K. (2011) Virulence of an emerging pathogenic lineage of Vibrio nigripulchritudo is dependant on two plasmids. Environ. Microbiol. 13 (2):296-306.
  19. Le Roux F, Davis B.M., Waldor M.K. (2011) A conserved small RNA governs replication and incompatability in diverse plasmids in marine bacteria. Nucleic Acids Res. 39 (3): 1004-13.
  20. O. Santos E, Alves N, Dias G, Mazotto AM, Vermelho A, Vora G, Wilson B, Beltran V, Bourne D, Le Roux F, Thompson F. (2011) Genomics and proteomics of the coral pathogen Vibrio coralliilyticus reveal a vast virulence repertoire. ISME J 5(9):1471-83.
  21. Cordero OX, Windschutte H, Kirkup B, Proehl S, Hussain F, Le Roux F, Mincer T,  Polz MF. Antibiotic production and resistance leads to social cohesion within bacterial populations in the wild. Science, September 2012.
  22. Labreuche Y, Pallandre L, Ansquer D, Herlin J, Wapotro B, Le Roux F. (2012) Pathotyping of Vibrio isolates by multiplex PCR reveals a risk of virulent strain spreading in New Caledonian shrimp farms. Microb Ecol. 63(1):127-38.
  23. Goudenège D, Labreuche Y., Krin E., Ansquer D., Mangenot S., Calteau A., Médigue C., Mazel D., Polz MF, Le Roux F. Comparative genomics of pathogenic lineages of Vibrio nigripulchritudo identifies virulence associated traits. ISMEJ, in press.