Evaluation of the production of paralytic shellfish poisoning toxins by extracellular bacteria isolated from the toxic dinoflagellate Alexandrium minutum.

The purpose of the study was to determine if paralytic shellfish poisoning (PSP) toxins are present in extracellular bacteria isolated from a toxic strain of the dinoflagellate Alexandrium minutum. A quantitative analysis was carried out of viable culturable bacteria attached to the surface of dinoflagellates and of bacteria present in dinoflagellate culture medium. A numerical taxonomy study was undertaken for presumptive identification of bacteria attached to the surface of dinoflagellates. Members of the following genera were detected on the cell surface of A. minutum: Cellulophaga, Marinomonas, Pseudoalteromonas, and Vibrio. The presence of intracellular PSP toxins in bacteria isolated from the cell surface of dinoflagellates was analysed by high-performance liquid chromatography with fluorescence detection (HPLC-FLD). Compounds that eluted at the same time as the standards of the PSP toxins GTX-2, GTX-3, GTX-4, dcGTX-2, and dcGTX-3 were present in some of the bacterial cell extracts. Natural fluorescent bacterial compounds, coeluting with some PSP toxins, were also detected. The results obtained showed that the fluorescent compounds, identified as putative PSP toxins by HPLC-FLD, did not correspond to any PSP analogue. This allowed us to reject the hypothesis that extracellular bacteria attached to the surface of dinoflagellates produce PSP toxins.

Effect of bacterial epiflora on egg hatching of the Atlantic sardine (Sardina pilchardus).

The aim of this work was to study the influence of bacterial epiflora on egg hatching of the sardine (Sardina pilchardus) obtained from a natural environment (Ría de Vigo, Spain) during the spawning season of the sardine (from January to June). Total bacteria, viable bacteria or the presence of specific potential pathogens for eggs, such as Pseudoalteromonas piscicida and Tenacibaculum (Flexibacter) ovolyticus, did not affect the viability of sardine eggs. Additionally, no relationship was observed between the presence of Vibrio spp., pathogenic for fish larvae, and the egg hatching. This was probably because the amount of bacteria associated with the eggs were between 10(2) and 10(4) orders lower than those found so far on the eggs of different fish species in rearing systems. Therefore, epiphytic bacteria did not affect the wild sardine eggs and, hence, in the area studied, it is probably not an important factor affecting annual recruitment success of this pelagic fish species.

Bacteria associated with sardine (Sardina pilchardus) eggs in a natural environment (Ría de Vigo, Galicia, northwestern Spain).

Abstract The present study was undertaken to describe the epiflora of the eggs of an important fishing species collected in a coastal zone. Microflora associated with sardine (Sardina pilchardus) eggs collected in the Ría de Vigo was examined from January to June 2000. The count was carried out in three different ways: a total direct count by epifluorescence, a heterotrophic bacteria count on marine agar (MA) and a total vibrio count on thiosulfate citrate bile sucrose (TCBS). It was observed that the counts of total bacteria by epifluorescence were always higher by 2-3 logarithms than the bacterial counts on MA, and by 3-4 logarithms than the count of vibrios on TCBS. In both cases the differences were statistically significant. Throughout the sampling period only a slight variation was observed in the counts undertaken, and in the measured physicochemical parameters. For the qualitative study, 250 strains isolated from MA and 81 strains recovered on TCBS were identified. Members of the genera Vibrio, Pseudoalteromonas, Pseudomonas and Moraxella were found to dominate on the culturable adherent microflora of sardine eggs and Aeromonas, Tenacibaculum (Flexibacter), Flavobacterium and Cytophaga spp. were present in minor amounts. Vibrio anguillarum and Vibrio fischeri, pathogens of fish larvae, as well as Tenacibaculum ovolyticum, a pathogen of fish eggs, were detected.