Stabili L. , C. Gravili, S. M. Tredici, F. Boero and P. Alifano, Association of a luminous Vibrio sp.

,
taxonomically related to Vibrio harveyi, with Clytia linearis (Thornely, 1900) (Hydrozoa, Cnidaria).
Journal of Experimental Marine Biology and Ecology. 2010. doi:10.1016/j.jembe.2010.10.004;

Loredana Stabili a, b, Cinzia Gravili b, Salvatore Maurizio Tredici b, Ferdinando Boero b and Pietro
Alifano b;

a Istituto Ambiente Marino Costiero, Sezione di Taranto, CNR, 74100 Taranto, Italy;

b Di.S.Te.B.A., University of Salento, 73100 Lecce, Italy;

Received 18 June 2010; revised 1 October 2010; accepted 4 October 2010. Available online 5
November 2010.

Abstract: A previously unknown association between a luminous Vibrio sp., taxonomically related to the
species Vibrio harveyi and a common member of the shallow/mid water communities of the
Mediterranean Sea, the hydrozoan Clytia linearis is described. All the specimens of C. linearis observed
under blue light excitation showed both a natural luminescence appearing as a series of fine dots due to
clytin, and a clear fluorescence on the external side of the perisarc around the colonies due to the
presence of luminous bacteria. Luminous bacteria were isolated from the surface of C. linearis, their
phenotypic characterization as isolates was performed by several morphological, biochemical, and
cultural tests, completed with 16S rDNA sequence analysis. All the isolates were referred to a Vibrio sp.
taxonomically related to V. harveyi. The association of the V. harveyi-related species with C. linearis, as
already suggested for another hydroid, Aglaophenia octodonta, could be explained with the activity of
these bacteria of feeding on the chitinous structures present in these hydroids. Moreover, the adhesion
of the luminous bacterium (here referred to as Vibrio sp. CL1) on C. linearis may contribute to the
survival of this Vibrio species in the marine environment providing a suitable growth habitat.

---------------------------------------------------------------------------------------------------------------------------------

Research Highlights:

►We report the association between a luminous Vibrio and the hydrozoan Clytia linearis. ►Luminous
bacteria were isolated from the surface of C. linearis. ►The characterization of the isolates was
performed by phenotypic–genotypic analyses. ►The luminous bacterium on C. linearis here is referred
to as Vibrio sp. CL1. ►The observed association has ecological as well as epidemiological significance.

From the text of the paper: “... Marine invertebrates and their associated microbiota provide a rich
source of material for studies that can reveal much about both the diversity in host–bacteria
associations and the mechanisms by which symbioses originate and persist ([Douglas, 1994] and
[Ostroumov, 2001]). ...”

---------------------------------------------------------------------------------------------------------------------------------

Keywords: Alien species; Bioluminescence; Clytia linearis; Epibiotic association; Hydrozoa; Vibrio harveyi;
Vibriosis; Vibrios distribution.

-----------------------------------------------------------------------------------------------------------------------------------------

Article Outline:

1. Introduction;
2. Materials and methods;

2.1. Sampling;

2.2. Microscope observation;

2.3. Quantitative analysis of bacteria living on C. linearis surface and phenotypic characterization of
luminous bacterial isolates;

2.4. 16S rRNA gene sequence analysis of bacterial isolates;

GenBank accession;

3. Results;

4. Discussion;

Acknowledgements;

References.

Fig. 1. Map showing the sampling station in the Northern Ionian Sea off the coast of Otranto, Lecce
(Italy).

--------------------------------------------------------------------------------

Fig. 2. Clytia linearis photomicrographs, living material: (a) colony at transmitted light; (b) colony at
epifluorescence. Scales: (a and b) 250 μm.

--------------------------------------------------------------------------------

Fig. 3. Phylogenetic analysis of 16S rRNA gene sequences of the luminous Vibrio species isolated from
Clytia linearis. Taxonomic position of Vibrio sp. CL1 is indicated with respect to several type strains. AO1,
VH1 and MT1 are Vibrio sp. strains previously isolated from the hydrozoans Aglaophenia octodonta and
Ventromma halecioides, and the bryozoan Myriapora truncata, respectively ([Stabili et al., 2006] and
[Stabili et al., 2008]). Bootstrap values ≥ 60 are reported at the branch points.

--------------------------------------------------------------------------------

Table 1.

Viable count of total culturable and luminous bacteria isolated from C. linearis colonies.

--------------------------------------------------------------------------------

Table 2.

The results of analysis of Vibrio sp. CL1 morphological and physiological properties.

r: rod shaped; d: diverse; F: fermentative; nd: no data; +: positive reaction; −: negative reaction; (−):
negative for 25–11%.

Corresponding author. Istituto Ambiente Marino Costiero, Sezione di Taranto, CNR, 74100 Taranto, Italy.
Note to users: The section "Articles in Press" contains peer reviewed accepted articles to be published in
this journal. When the final article is assigned to an issue of the journal, the "Article in Press" version will
be removed from this section and will appear in the associated published journal issue. The date it was
first made available online will be carried over. Please be aware that although "Articles in Press" do not
have all bibliographic details available yet, they can already be cited using the year of online publication
and the DOI as follows: Author(s), Article Title, Journal (Year), DOI. Please consult the journal's reference
style for the exact appearance of these elements, abbreviation of journal names and the use of
punctuation.

There are three types of "Articles in Press":

Accepted manuscripts: these are articles that have been peer reviewed and accepted for publication by
the Editorial Board. The articles have not yet been copy edited and/or formatted in the journal house
style.

Uncorrected proofs: these are copy edited and formatted articles that are not yet finalized and that will
be corrected by the authors. Therefore the text could change before final publication.

Corrected proofs: these are articles containing the authors' corrections and may, or may not yet have
specific issue and page numbers assigned.

--------------------------------------------------------------------------------

Journal of Experimental Marine Biology and Ecology; Article in Press, Corrected Proof - Note to users;

Bibliography of other relevant publications (Ecology, Environment, Aquatic Sciences, Ecosystems,

Biosphere, Biology):
http://www.researchgate.net/profile/Sergei_Ostroumov/blog/3380_Ecology_biology_life_sciences_Bibliography;