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Applied and Environmental Microbiology, July 2003, p. 3932-3937, Vol. 69, No. 7
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.7.3932-3937.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Dominance of Vibrio fischeri in Secreted Mucus outside the Light Organ of Euprymna scolopes: the First Site of Symbiont Specificity

Spencer V. Nyholm{dagger} and Margaret J. McFall-Ngai*

Pacific Biomedical Research Center, Kewalo Marine Laboratory, University of Hawaii, Honolulu, Hawaii 96813

Received 20 December 2002/ Accepted 10 April 2003

Previous studies of the Euprymna scolopes-Vibrio fischeri symbiosis have demonstrated that, during colonization, the hatchling host secretes mucus in which gram-negative environmental bacteria amass in dense aggregations outside the sites of infection. In this study, experiments with green fluorescent protein-labeled symbiotic and nonsymbiotic species of gram-negative bacteria were used to characterize the behavior of cells in the aggregates. When hatchling animals were exposed to 103 to 106 V. fischeri cells/ml added to natural seawater, which contains a mix of approximately 106 nonspecific bacterial cells/ml, V. fischeri cells were the principal bacterial cells present in the aggregations. Furthermore, when animals were exposed to equal cell numbers of V. fischeri (either a motile or a nonmotile strain) and either Vibrio parahaemolyticus or Photobacterium leiognathi, phylogenetically related gram-negative bacteria that also occur in the host's habitat, the symbiont cells were dominant in the aggregations. The presence of V. fischeri did not compromise the viability of these other species in the aggregations, and no significant growth of V. fischeri cells was detected. These findings suggested that dominance results from the ability of V. fischeri either to accumulate or to be retained more effectively within the mucus. Viability of the V. fischeri cells was required for both the formation of tight aggregates and their dominance in the mucus. Neither of the V. fischeri quorum-sensing compounds accumulated in the aggregations, which suggested that the effects of these small signal molecules are not critical to V. fischeri dominance. Taken together, these data provide evidence that the specificity of the squid-vibrio symbiosis begins early in the interaction, in the mucus where the symbionts aggregate outside of the light organ.

* Corresponding author. Mailing address: Pacific Biomedical Research Center, Kewalo Marine Laboratory, University of Hawaii, 41 Ahui St., Honolulu, HI 96813. Phone: (808) 539-7310. Fax: (808) 599-4817. E-mail: mcfallng{at}hawaii.edu.

{dagger} Present address: Stanford University, Hopkins Marine Station, Pacific Grove, CA 93950.

Applied and Environmental Microbiology, July 2003, p. 3932-3937, Vol. 69, No. 7
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.7.3932-3937.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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