1887

Abstract

A group of seven Chilean isolates presumptively belonging to Vibrio tapetis was isolated from diseased fine flounders (Paralichthys adspersus) and red conger eel (Genypterus chilensis) experimentally reared in Quintay (Chile). All isolates were confirmed as members of V. tapetis on the basis of matrix-assisted laser desorption ionization time-of-flight MS, 16S rRNA gene sequencing, DNA–DNA hybridization values and G+C content. The ERIC-PCR and REP-PCR patterns were homogeneous among those isolates recovered from the same host (red conger or fine flounders), but distinct from the type strains V. tapetis subsp. tapetis CECT 4600 and V. tapetis subsp. britannicus CECT 8161. On the basis of atpA, rpoA, rpoD, recA and pyrH gene sequence similarities (99.7–100 %) and clustering in the phylogenetic trees, the red conger isolates (Q20, Q047, Q48 and Q50) were confirmed as representing V. tapetis subsp. tapetis . However, they differed from V. tapetis subsp. tapetis CECT 4600 in their lipase, alpha quimiotripsin and non-acid phosphatase production. On the other hand, the fine flounder isolates (QL-9, QL-35 and QL-41) showed rpoD, recA and pyrH gene sequence similarities ranging from 91.6 to 97.7 % with the type strains of the two V. tapetis subspecies (CECT 4600 and CECT 8161) and consistently clustered together as an independent phylogenetic line within V. tapetis . Moreover, they could be differentiated phenotypically from strains CECT 4600 and CECT 8161 by nine and three different biochemical tests, respectively. In conclusion, the presence of V. tapetis in diseased red conger eel and fine flounder was demonstrated, extending the known host range and geographical location for this pathogen. Furthermore, this study demonstrates that the three isolates from fine flounder represent a novel subdivision within V. tapetis , for which the name V. tapetis subsp. quintayensis subsp. nov. is proposed and with QL-9 (=CECT 8851=LMG 28759) as the type strain. Although QL-9 was isolated from kidney of diseased fine flounder specimens, the challenge assays showed that it was non-pathogenic for this species.

Loading

Article metrics loading...

/content/journal/ijsem/10.1099/ijsem.0.001705
2017-04-03
2019-10-15
Loading full text...

Full text loading...

/deliver/fulltext/ijsem/67/3/716.html?itemId=/content/journal/ijsem/10.1099/ijsem.0.001705&mimeType=html&fmt=ahah

References

  1. Paillard C, Maes P, Oubella R. Brown ring disease in clams. Annu Rev Fish Dis 1994;4:219–240 [CrossRef]
    [Google Scholar]
  2. Paillard C, Percelay L, Le Pennec M, Picard DL. Origine pathogene de l’anneau brun chez Tapes phillipinarum (Mollusque, ivalve). CR Acad Sci 1989;309:235–241
    [Google Scholar]
  3. Borrego JJ, Castro D, Luque A, Paillard C, Maes P et al. Vibrio tapetis sp. nov., the causative agent of the brown ring disease affecting cultured clams. Int J Syst Bacteriol 1996;46:480–484 [CrossRef]
    [Google Scholar]
  4. Reid HI, Duncan HL, Laidler LA, Hunter D, Birkbeck TH. Isolation of Vibrio tapetis from cultivated Atlantic halibut (Hippoglossus hippoglossus L.). Aquaculture 2003;221:65–74 [CrossRef]
    [Google Scholar]
  5. Jensen S, Samuelsen OB, Andersen K, Torkildsen L, Lambert C et al. Characterization of strains of Vibrio splendidus and V. tapetis isolated from corkwing wrasse Symphodus melops suffering vibriosis. Dis Aquat Organ 2003;53:25–31 [CrossRef][PubMed]
    [Google Scholar]
  6. López JR, Balboa S, Núñez S, de La Roca E, de La Herran R et al. Characterization of Vibrio tapetis strains isolated from diseased cultured wedge sole (Dicologoglossa cuneata Moreau). Res Vet Sci 2011;90:189–195 [CrossRef][PubMed]
    [Google Scholar]
  7. Declercq AM, Chiers K, Soetaert M, Lasa A, Romalde JL et al. Vibrio tapetis isolated from vesicular skin lesions in Dover sole Solea solea. Dis Aquat Organ 2015;115:81–86 [CrossRef][PubMed]
    [Google Scholar]
  8. Rodríguez JM, López-Romalde S, Beaz R, Alonso MC, Castro D et al. Molecular fingerprinting of Vibrio tapetis strains using three PCR-based methods: ERIC-PCR, REP-PCR and RAPD. Dis Aquat Organ 2006;69:175–183 [CrossRef][PubMed]
    [Google Scholar]
  9. Balboa S, Romalde JL. Multilocus sequence analysis of Vibrio tapetis, the causative agent of brown ring disease: description of Vibrio tapetis subsp. britannicus subsp. nov. Syst Appl Microbiol 2013;36:183–187 [CrossRef][PubMed]
    [Google Scholar]
  10. Valdebenito S, Avendaño-Herrera R. Phenotypic, serological and genetic characterization of Flavobacterium psychrophilum strains isolated from salmonids in Chile. J Fish Dis 2009;32:321–333 [CrossRef][PubMed]
    [Google Scholar]
  11. Pazos F, Santos Y, Macias AR, Nunez S, Toranzo AE. Evaluation of media for the successful culture of Flexibacter maritimus. J Fish Dis 1996;19:193–197 [CrossRef]
    [Google Scholar]
  12. Clinical and Laboratory Standard Institute (CLSI) Methods for Antimicrobial Disk Susceptibility Testing of Bacteria Isolated from Aquatic Animals. Approved Guideline. CLSI Document M42-A (ISBN 1-56238-611-5) Wayne, Pennsylvania: Clinical and Laboratory Standard Institute; 2006
    [Google Scholar]
  13. Versalovic J, Koeuth T, Lupski JR. Distribution of repetitive DNA sequences in eubacteria and application to fingerprinting of bacterial genomes. Nucleic Acids Res 1991;19:6823–6831 [CrossRef][PubMed]
    [Google Scholar]
  14. Maier T, Klepel S, Renner U, Kostrzewa M. Fast and reliable MALDI-TOF MS–based microorganism identification. Nat Methods 2006;3: [CrossRef]
    [Google Scholar]
  15. Edwards U, Rogall T, Blöcker H, Emde M, Böttger EC. Isolation and direct complete nucleotide determination of entire genes. characterization of a gene coding for 16S ribosomal RNA. Nucleic Acids Res 1989;17:7843–7853 [CrossRef][PubMed]
    [Google Scholar]
  16. Kim O-S, Cho Y-J, Lee K, Yoon S-H, Kim M et al. Introducing EzTaxon-e: a prokaryotic 16S rRNA gene sequence database with phylotypes that represent uncultured species. Int J Syst Evol Microbiol 2012;62:716–721 [CrossRef]
    [Google Scholar]
  17. Tamura K, Peterson D, Peterson N, Stecher G, Nei M et al. MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol 2011;28:2731–2739 [CrossRef][PubMed]
    [Google Scholar]
  18. Kimura M. A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences. J Mol Evol 1980;16:111–120 [CrossRef][PubMed]
    [Google Scholar]
  19. Saitou N, Nei M. The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 1987;4:406–425[PubMed]
    [Google Scholar]
  20. Gonzalez JM, Saiz-Jimenez C. A fluorimetric method for the estimation of G+C mol% content in microorganisms by thermal denaturation temperature. Environ Microbiol 2002;4:770–773 [CrossRef][PubMed]
    [Google Scholar]
  21. Lasa A, Avendaño-Herrera R, Estrada JM, Romalde JL. Isolation and identification of Vibrio toranzoniae associated with diseased red Conger eel (Genypterus chilensis) farmed in Chile. Vet Microbiol 2015;179:327–331 [CrossRef][PubMed]
    [Google Scholar]
  22. Urdiain M, López-López A, Gonzalo C, Busse HJ, Langer S et al. Reclassification of Rhodobium marinum and Rhodobium pfennigii as Afifella marina gen. nov. comb. nov. and Afifella pfennigii comb. nov., a new genus of photoheterotrophic Alphaproteobacteria and emended descriptions of Rhodobium, Rhodobium orientis and Rhodobium gokarnense. Syst Appl Microbiol 2008;31:339–351 [CrossRef][PubMed]
    [Google Scholar]
  23. Avendaño-Herrera R. Enfermedades infecciosas del cultivo de salmónidos en Chile y el mundo Puerto Varas: NIVA; 2011
    [Google Scholar]
  24. Tapia-Cammas D, Yañez A, Arancibia G, Toranzo AE, Avendaño-Herrera R. Multiplex PCR for the detection of Piscirickettsia salmonis, Vibrio anguillarum, Aeromonas salmonicida and Streptococcus phocae in Chilean marine farms. Dis Aquat Organ 2011;97:135–142 [CrossRef][PubMed]
    [Google Scholar]
  25. Balboa S, Bastardo A, Romalde JL. Disentangling the population structure and evolution of the clam pathogen Vibrio tapetis. Microb Ecol 2014;67:145–154 [CrossRef][PubMed]
    [Google Scholar]
  26. Sawabe T, Ogura Y, Matsumura Y, Feng G, Amin AR et al. Updating the Vibrio clades defined by multilocus sequence phylogeny: proposal of eight new clades, and the description of Vibrio tritonius sp. nov. Front Microbiol 2013;4:414 [CrossRef][PubMed]
    [Google Scholar]
  27. Stackebrandt E, Goebel BM. Taxonomic note: a place for DNA-DNA reassociation and 16S rRNA sequence analysis in the present species definition in bacteriology. Int J Syst Evol Microbiol 1994;44:846–849 [CrossRef]
    [Google Scholar]
  28. Fuentes EN, Ruiz P, Valdes JA, Molina A. Catabolic signaling pathways, atrogenes, and ubiquitinated proteins are regulated by the nutritional status in the muscle of the fine flounder. PLoS One 2012;7:e44256 [CrossRef][PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/ijsem/10.1099/ijsem.0.001705
Loading
/content/journal/ijsem/10.1099/ijsem.0.001705
Loading

Data & Media loading...

Supplements

Supplementary File 1

PDF

Most Cited This Month

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error