Yolaine Delgado Gómez, Rodolfo Umaña Castro, Stefany Solano González, María V. Iglesias Rodríguez, Eudalys Ortiz Guilarte, Carlos Álvarez Valcárcel, Gladys M. Lugioyo Gallardo


Light emitted by luminescent bacteria is sensitive to several toxic compounds; therefore, some luminous species have been used to evaluate the quality of water environments. In the present study we carried out the phenotypic characterization and molecular identification of a marine luminescent isolate (CBM-784) from the NW Cuban coast. The identification of the CBM-784 luminous isolate was based on phenotypic and genotypic characteristics. Phenotypically, the CBM-784 strain revealed the following characteristics: Gram negative, positive to oxidase and catalase reactions,
bioluminescent, and facultative anaerobic respiration. The bacterial strain produces enzymes with gelatinase, lysine carboxylase and amylase activity. Taken together, these assays indicated that CBM-784 showed a high phenotypic similarity to the Vibrio harveyi ATCC 14126 strain. On the basis of 16S rRNA gene sequencing, CBM-784 was closely related to Vibrio harveyi and Vibrio rotiferianus (94% similarity). Sequence analysis of gyrB gene, has shown that CBM-784 shares taxonomic position with Vibrio campbellii and Vibrio harveyi isolates, with 95% of bootstrap value. In addition, sequence analysis of pyrH gene, grouped this isolate to the Vibrio harveyi cluster with a strong bootstrap support (99%). The multilocus sequence analysis and phenotypic characterization of CBM-784 indicated that this strain have a strong relation to Vibrio harveyi.

Palabras clave

luminescent bacteria; Vibrionaceae; Vibrio harveyi; Mutilocus sequence analysis

Texto completo:



Alsina, M., Blanch, A.R. 1994. A set of keys for biochemical identification of environmental Vibrio species. Journal of Applied Bacteriology, 76: 79-85.

Altschul, S.F., Gish, W., Miller, W., Myers, E.W., Lipman, D.J. 1990. Basic local alignment search tool. Journal of Molecular

Biology, 21: 403-410.

Ast, J.C., Dunlap, P.V. 2005. Phylogenetic resolution and habitat specificity of the Photobacterium phosphoreum species group. Environmental Microbiology, 7: 1641-1654.

Bagordo, F., Serio, F., Lugoli, F., Grassi, T., Idolo, A., Gabutti, G., De Donno, A. 2012. Phenotypic characterization of culturable marine luminous bacteria isolated from coastal waters of the southern Adriatic Sea (Otranto, Italy). Ciencias Marinas, 38: 599-608.

Balcázar, J.L., Vendrell, D., de Blas, I., Ruiz-Zarzuela, I., Girone´s, O., Mu´zquiz, J.L. 2007. Quantitative detection of Aeromonas salmonicida in fish tissue by real-time PCR using selfquenched, fluorogenic primers. Journal of Medical Microbiology, 56: 323-328.

Baumann, P., Baumann, L. 1981. The marine Gram-negative eubacteria: genera Photobacterium, Beneckea, Alteromonas, Pseudomonas, & Alcaligenes. The Prokaryotes. A Handbook on Habitats, Isolation, & Identification of Bacteria. Starr, M.P., Stolp, H., Trüper, H.G., Balows, A., Schlegel, H.G. (eds.), pp 1302–1331. Springer-Verlag, Berlin.

Cano-Gómez, A., Høj, L., Owens, L., Andreakis, N. 2011. Multilocus sequence analysis provides basis for fast and reliable identification of Vibrio harveyi-related species and reveals previous misidentification of important marine pathogens. Syst Appl Microbiol, 34: 561-565.

Cano-Gómez, A., Høj, L., Owens, L., Baillie, B.K., Andreakis, N. 2015. A multiplex PCR-based protocol for identification and quantification of Vibrio harveyi-related species. Aquaculture, 437: 195-200.

Chiu, H.-H., Chou, H.-H., Jean, W.-D., Shieh, W.-Y. 2007. Isolation and Characterization of marine luminous bacteria from shallow coastal waters of Taiwan. Journal of Microbiology, Immunology and Infection, 40: 14-23.

Drews, G. 2000. The roots of microbiology and the influence of Ferdinand Cohn on microbiology of the 19th century. FEMS

Microbiol Rev, 24: 225-249.

Dunlap, P.V., Urbanczyk, H. 2013. Luminous bacteria. The Prokaryotes. Springer Berlin Heidelberg. Dupy, B., Daube, G., Popoff, M.R., Colem, S.T. 1997. Clostridium perfringens urease genes are plasmid borne. InfecImmun, 65: 2313-2320.

Edgar, R.C. 2010. Search and clustering orders of magnitude faster than BLAST. Bioinformatics, 26: 2460-2461.

Farto, R., Montes, M., Pérez, M.J., Nieto, T.P., Larsen, J.L., Pedersen, K. 1999. Characterization by numerical taxonomy and ribotyping of Vibrio splendidus biovar I and Vibrio scophthalmi strains associated with turbot cultures. J. Appl. Bacteriol, 86: 796-804.

Girotti, S., Ferri, E.N., Fumo, M.G., Maiolini, E. 2008. Monitoring of environmental pollutants by bioluminescent bacteria (Review). Analytica Chimica Acta Article: 2-29.

Gómez-Gil, B., Roque, A., Chimetto, L., Moreira, A.P.B., Lang, E., Thompson, F.L. 2012. Vibrio alfansensis sp. nov., isolated from marine organisms. International Journal of Systematic and Evolutionary Microbiology, 62: 2955-2961.

Harrigan, W.F., McCance, M.E. 1968. Métodos de laboratorio en microbiología. pp 426. Editorial Academia, España.

Ishiguro, N., Sato, G. 1979. The distribution of plasmids determining citrate utilization in citrate-positive variants of Escherichia coli from humans, domestic animals, feral birds and environments. J. Hyg, 83: 331-237.

Lal, M.T.B.M., Ransangan, J. 2013. Taxonomic classification of Vibrio harveyi using 16S rDNA and atpA gene sequencing method. International Journal of Research in Pure and Applied Microbiology, 3: 17- 24.

Lane, D.J. 1991. 16S/23S rRNA sequencing. In nucleic acid techniques in bacterial systematics. Goodfellow, E.S.M. (ed.)

pp. Wiley, New York.

López-Caballero, M.E., Torres, M.D.A., Sánchez-Fernández, J.A., Moral, A. 2002. Photobacterium phosphoreum isolated as a luminescent colony from spoiled fish, cultured in a model system under controlled atmospheres. Eur. Food Res. Technol, 215: 390-395.

Lunder, T., Sorum, H., Holstad, G., Steigerwalt, A.G., Mowinckel, P., J, B.D. 2000. Phenotypic and genotypic characterization of Vibrio viscosus sp. nov. and Vibrio wodanis sp. nov., isolated from Altantic salmon (Salmo salar) with ‘winter ulcer’. Int. J. Syst. Evol. Microbiol, 50: 427-450.

Makemson, J.C., Fulayfil, N.R., Landry, W., Van Ert, L.M., Wimpee, C.F., Widder, E.A., Case, J.F. 1997.

Shewanella woodyi sp. nov., an exclusively respiratory luminous bacterium isolated from the Alboran Sea. Int. J. Syst. Bacteriol, 47: 1034-1039.

Mohammad, T., Mohamad, B., Ransangan, J. 2013. Taxonomic

classification of Vibrio harveyi using 16S rDNA and atpA gene sequencing method. International Journal of Research in Pure and Applied Microbiology, 3: 17-24.

Nealson, K.H., Hastings, J.W. 1979. Bacterial bioluminescence: its control and ecological significance. Microbiol Rev, 43: 496-518.

Pascual, J., Macián, M.C., Arahal, D.R., Garay, E., Pujalte, M.J. 2010. Multilocus sequence analysis of the central clade of the

genus Vibrio by using the 16S rRNA, recA, pyrH, rpoD, gyrB, rctB and toxR genes. International Journal of Systematic and Evolutionary Microbiology, 60: 154-165.

Podgórska, B., K, P., Pempkowiak, P., Wegrzyn, G. 2007. The use of a novel Vibrio harveyi luminescence mutagenicity assay in testing marine water for the presence of mutagenic pollution. Marine Pollution Bulletin, 54: 792-819.

Podgórska, B., Węgrzyn, G. 2007. A Modified Vibrio harveyi mutagenicity assay based on bioluminescence induction. Letters in Applied Microbiology, 42: 578-582.

Rambaut, A. 2009. FigTree v1.4. 2012-2014. Available via http:// [Online]. Ransangan, J., Mustafa, S. 2009. Identification of Vibrio harveyi isolated from diseased asian seabass lates calcarifer by use of 16s ribosomal DNA sequencing. Journal of Aquatic Animal Health, 21: 150-155.

Sáenz, C.I., Nevárez, G.V. 2010. La bioluminiscencia de

microorganismos marinos y su potencial biotecnológico.

Revista Científica de la Universidad Autónoma de Coahuila,

: 1-7.

Satomi, M., Oikawa, H., Yano, Y. 2003. Shewanella marinintestina sp. nov., Shewanella schlegeliana sp. nov. and Shewanella sairae sp. nov., novel eicosapentaenoic-acid-producing marine bacteria isolated from sea-animal intestines. Int. J. Syst. Evol. Microbiol, 53: 491-499.

Shanware, A., Thakre, N., Pande, S. 2013. Isolation and characterization of novel marine luminescent bacteria from Diu beach, India. Journal of Pharmacy Research, 7: 529-533. Silvestro, D., Michalak, I. 2012. raxmlGUI: a graphical front-end for RAxML. Org. Divers. Evol, 12: 335-337.

Stamatakis, A., Ludwig, T., Meier, H. 2005. RAxML-III: a fast program for maximum likelihood-based inference of large phylogenetic trees. Bioinformatics, 21: 456-463.

Thompson, F.L., Gevers, D., Thompson, C.C., Dawyndt, P., Naser, S., Hoste, B., Munn, C.B., Swings, J. 2005. Phylogeny and molecular identification of Vibrios on the basis of multilocus sequence analysis. Applied and Environmental Microbiology, 71: 5107-5115.

Urbanczyk, H., Ogura, Y., Hayashi, T. 2013. Taxonomic revision of Harveyi clade bacteria (family Vibrionaceae) based on analysis of whole genome sequences. International Journal of Systematic and Evolutionary Microbiology, 63: 2742-2751.

Urbanczyk, H., Ogura, Y., Hayashi, T. 2014. Contrasting inter- and intraspecies recombination patterns in the “Harveyi clade” vibrio collected over large spatial and temporal scales. Rev. Genome Biol. Evol, 7: 71-80.

Vandamme, P., Pot, B., Gillis, M., De Vos, P., Kersters, K., Swings, J. 1996. Polyphasic taxonomy, a consensus approach to bacterial systematics. Microbiol Rev, 60: 407-438.

Venkateswaran, K., Moser, D.P., Dollhopf, M.E., Lies, D.P., Saffarini, D.A., MacGregor, B.J., Ringelberg, D.B., White, D.C., Nishijima, M., Sano, H., Burghardt, J., Stackebrandt, E., Nealson, K.H. 1999. Polyphasic taxonomy of the genus Shewanella and description of Shewanella oneidensis sp. nov. Int. J. Syst. Bacteriol, 49: 705-724.


Enlaces refback

  • No hay ningún enlace refback.
 |  Añadir comentario