The microbiota of the body wall lesions of the echinoid Tripneustes gratilla, initiated by the grazing action of the parasitic gastropod Vexilla vexillum, was investigated with a pluridisciplinary approach. Parasitised sea urchins showed body wall lesions strongly infected by bacteria that progressed through the test and reached the coelomic cavity after ca. 1 mo. We report here on the bacterial community observed in lesions of echinoids collected in situ and on the bacteria that successively appeared during laboratory experiments. Two Alphaproteobacteria, a CFB (Cytophaga-Flavobacterium-Bacteroides) bacterium, 3 Vibrio species and Exiguobacterium aestuarii were identified in the field-collected lesions by 16S rDNA sequencing. The last 4 bacteria were cultured and each induced the disease when inoculated on scalpel-made wounds, with 100% of the individuals infected within 2 d. Scalpel-induced scarifications tended to heal within 3 wk, while gastropod-induced lesions evolved into disease, suggesting a role of Vexilla vexillum in the development of the infection. Denaturing gradient gel electrophoresis (DGGE) and sequencing suggest that (1) bacteria associated with healthy integument were not present in the lesions and were thus not responsible for their infection, (2) Alphaproteobacteria with close phylogenetic affiliation with other bacteria involved in several diseases affecting marine invertebrates were present, and (3) these Alphaproteobacteria were present from the beginning of the infection and appeared earlier in the infection than other bacteria such as CFB bacteria.
KEY WORDS: Echinoderm · DGGE · Bacteria · Infection · Lesion · Gastropod
Resale or republication not permitted without written consent of the publisherDis Aquat Org 77: [73][74][75][76][77][78][79][80][81][82] 2007 gastropod grazes the body surface, which progressively turns black as the infection by microorganisms develops. Infections then progress through the test, often causing fatal perforations (Vaïtilingon et al. 2004). The present study examines this infection that leads to the death of T. gratilla. Microscopic, microbiological and molecular techniques were used to characterise and identify the microbial community associated with the lesions. The role of the gastropod in the infection is also investigated. This is the first report of a bacterial infection affecting sea urchins in the Indian Ocean. (Linnaeus, 1758) and Vexilla vexillum (Gmelin, 1791) individuals were collected by hand at low tide on the barrier reef off Toliara, Madagascar, from January to February 2004 and in February 2005. The collection site, Ambatobe (23°25' 00'' S, 43°39' 23'' E), was located on the boulder tract of the seaward slope of the reef flat. Eight lesions (named 'field-collected lesions' hereafter) were sampled on the reef, of which 4 were fixed in non-acetic Bouin's fluid for scanning electron microscopy (SEM) and 4 in absolute ethanol for molecular analyses. Other specimens were brought to the laboratory and left in aquaria with running s...