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Marine mollusc shell growth has been widely measured using fluorochrome marking. In order to test the efficiency and reliability of calcein staining on Pinctada margaritifera shells and pearls, the present study examined two administration methods, different concentrations and several immersion times. Immersion in a 150 mg L−1 calcein solution for 12 h to 24 h appeared to be the best method for marking P. margaritifera shells. For pearl marking, injection of a 200 mg L−1 calcein solution into the pearl pouch was the optimal method. Calcein marking was then used to measure the influence of food resource levels on the shell growth. Groups of 23-month-old P. margaritifera were fed at three trophic levels for two months. The two highest food levels tested (6000 cell mL−1 and 15 000 cell mL−1) induced uniform growth between the dorsal and ventral sides of shell, whereas the lowest food level (800 cell mL−1) induced greater growth on the dorsal side. Shell deposits from the ventral side were observed using a scanning electron microscope, revealing that the difference of the trophic level over two months had modified the thickness of the aragonite tablets formed. These results showed that the trophic level is a major factor conditioning P. margaritifera development.

Bacterial populations association with phytoplankton cultures used as food for bivalve larvae were enumerated and identified from their partial 16S rDNA gene sequences. Microalgae were provided from different European hatcheries during the larval production season. Average concentration (direct counts) of bacteria ranged from 1.3 x 10(5) to 5.3 x 10(8) mL(-1) while culturable bacteria represented from 10% to >60% of total bacteria. In most cases, three to six representatives of each type of colony were collected on solid medium. The identity of isolates from the same colony type was checked by two different randomly amplified polymorphic DNA (RAPD) typing methods, after which the 16S rDNA gene of one to three isolates by colony type were partially sequenced. Algae harbored a large spectrum of bacteria belonging to the alpha-Proteobacteria, beta-Proteobacteria, gamma-Proteobacteria, Cytophaga- Flavobacterium- Bacteroides (CFB) group, Actinobacteria, and Bacillus. Members of the Roseobacter clade and CFB group were the most abundant. In the majority of cases one strain constituted 50% or more of the culturable bacterial flora. About half of the isolates were common to two hatcheries or at least two microalgal cultures. Several isolates were closely related to bacteria associated with harmful dinoflagellates in culture. Thus, the algal cultures seemed to favor certain bacterial species which belonged to distantly separated groups. As some of them could disturb the development of bivalve larvae, the control of bacterial populations would undoubtedly make it possible to reduce larval losses in bivalve rearing.