Free-living (FL) and particle-attached (PA) bacterial assemblages in the Northwest Mediterranean Sea were studied using pyrosequencing data of the 16S rRNA. We have described and compared the richness, the distribution of Operational Taxonomic Units (OTUs) within the two fractions, the spatial distribution, and the taxonomic composition of FL and PA bacterial assemblages. The number of OTUs in the present work was two orders of magnitude higher than in previous studies. Only 25% of the total OTUs were common to both fractions, whereas 49% OTUs were exclusive to the PA fraction and 26% to the FL fraction. The OTUs exclusively present in PA or FL assemblages were very low in abundance (6% of total abundance). Detection of the rare OTUs revealed the larger richness of PA bacteria that was hidden in previous studies. Alpha-Proteobacteria dominated the FL bacterial assemblage and gamma-Proteobacteria dominated the PA fraction. Bacteroidetes were important in the PA fraction mainly at the coast. The high number of sequences in this study detected additional phyla from the PA fraction, such as Actinobacteria, Firmicutes, and Verrucomicrobia.

No abstract

Paralytic Shellfish Poisoning (PSP) toxins are annually recurrent along the Massachusetts coastline (USA), which includes many small embayments and salt ponds. Among these is the Nauset Marsh System (NMS), which has a long history of PSP toxicity. Little is known, however, about the bloom dynamics of the causative organism Alexandrium fundyense within that economically and socially important system. The overall goal of this work was to characterize the distribution and dynamics of A. fundyense blooms within the NMS and adjacent coastal waters by documenting the distribution and abundance of resting cysts and vegetative cells. Cysts were found predominantly in three drowned kettle holes or salt ponds at the distal ends of the NMS – Salt Pond, Mill Pond, and Town Cove. The central region of the NMS had a much lower concentration of cysts. Two types of A. fundyense blooms were observed. One originated entirely within the estuary, seeded by cysts in the three seedbeds. These blooms developed independently of each other and of the A. fundyense population observed in adjacent coastal waters outside the NMS. The temporal development of the blooms was different in the three salt ponds, with initiation differing by as much as 30 days. These differences do not appear to reflect the initial cyst abundances in these locations, and may simply result from higher cell retention and higher nutrient concentrations in Mill Pond, the first site to bloom. Germination of cysts accounted for a small percentage of the peak cell densities in the ponds, so population size was influenced more by the factors affecting growth than by cyst abundance. Subsurface cell aggregation (surface avoidance) limited advection of the vegetative A. fundyense cells out of the salt ponds through the shallow inlet channels. Thus, the upper reaches of the NMS are at the greatest risk for PSP since the highest cyst abundances and cell concentrations were found there. After these localized blooms in the salt ponds peaked and declined, a second, late season bloom occurred within the central portions of the NMS. The timing of this second bloom relative to those within the salt ponds and the coastal circulation patterns at that time strongly suggest that those cells originated from a regional A. fundyense bloom in the Gulf of Maine, delivered to the central marsh from coastal waters outside the NMS through Nauset Inlet. These results will guide policy decisions about water quality as well as shellfish monitoring and utilization within the NMS and highlight the potential for “surgical” closures of shellfish during PSP events, leaving some areas open for harvesting while others are closed.

Dinoflagellate blooms in coastal upwelling systems are restricted to times and places with reduced exchange and mixing. The Rías Baixas of Galicia are four bays in the NW Iberian upwelling with these characteristics where harmful algal blooms (HABs) of dinoflagellates are common. These blooms are especially recurrent at the end of the upwelling season, when autumn downwelling amplifies accumulation and retention through the development of a convergence front in the interior of Rías. Because oceanic water enters the Rías during downwelling, it has been hypothesised that dinoflagellate blooms originate by the advection and subsequent accumulation of allochthonous populations. To examine this possibility, we studied the microplankton succession in relation to hydrographic variability in the Ría de Vigo (one of these four bays) along an annual cycle making use of a high sampling frequency. The results indicated that upwelling lasted from May to August, with downwelling prevailing in winter. Microplankton during upwelling, although dominated by diatoms, evidenced a progressive increase in the importance of dinoflagellates, which achieved maximum abundance at the end of the upwelling season. Thus, diatoms characterised the spring bloom, while diatoms and autochthonous dinoflagellates composed the autumn bloom. Diatoms dominated during the first moments of the autumn downwelling and dinoflagellates were more abundant later, after stronger downwelling removed diatoms from the water column. Since the dinoflagellates selected by downwelling belonged to the local community, it is concluded that advection of alien populations is not required to explain these autumn blooms. #

No abstract

The dilution technique, combined with identification and enumeration of pico-, nanoand microplankton by microscopy, was used to estimate microzooplankton impact on the microbial community in surface waters of a coastal embayment on the NW Iberian upwelling system. Microzooplankton were important consumers of autotrophic and heterotrophic plankton in this system, feeding up to 93% of standing stock and more than 100% of production of several groups. Heterotrophic bacteria and heterotrophic picoflagellates experienced the highest and constant impact, with 75-84% of their standing stocks and 85-102% of their production being channelled through the microbial food web. Pico-and nanophytoplankton were also consumed, although maximum grazing occurred on diatoms during upwelling events, coinciding with highest primary production. Predation on pico-nanoheterotrophs was especially relevant under downwelling conditions, when consumption of total carbon and particularly autotrophic carbon was considerably lower than during upwelling. The results suggest that the existence of a multivorous food web, extending from the microbial loop to the herbivorous food web, could be a major feature in this coastal upwelling system. The microbial loop, which occurs as a permanent background in the system, would contribute to sustain the microbial food web during downwelling, whereas the herbivorous food web could coexist with a microbial food web based on large diatoms during upwelling. The multivorous food web would partially divert diatoms from sinking and hence favour the retention of organic matter in the water column. This could enhance the energy transfer to higher pelagic trophic levels in coastal upwelling systems.

Size-fractionated phytoplankton was studied using pigments and carbon (C) biomass derived from light microscopy at a fixed location on the NW Iberian shelf between May 2001 and June 2002. The annual contributions of pico-, nano-and microphytoplankton to the total autotrophic C biomass were 9.7, 63 and 27.3%, respectively, with unidentified nanoflagellates (63%) and diatoms (22%) being the most important components. The CHEMTAX program was used to determine the chlorophyll a (chl a) contribution of 9 pigment groups in nano-microplankton and 8 in picoplankton. 'Diatoms II' (species containing chl c 3 ), 'haptophytes II' (species of the genus Chrysochromulina containing chl c 2 -MGDG [monogalactosyl diacylglyceride] [14:0/14:0] ) and 'chlorophytes', represented 34, 17 and 16%, respectively, of the total chl a in the nano-microplankton fraction. The major pigment groups in the picoplankton were 'chlorophytes', 'Synechococcus' and 'prasinophytes II' (Order Mamiellales), which accounted for 34, 22 and 10% of the total chl a in this size class. 'Prochlorococcus' was traced using divinyl (DV) chl a and was present seasonally, especially during downwelling and stratified poleward conditions (October to November). The lowest chl a and autotrophic C values throughout the study (152 ± 45 ng chl a l -1 and 31 ± 13 µg C l -1 ) corresponded to the homogenous poleward flow or Iberian Poleward Current (IPC) from December to February. During this period the phytoplankton composition changed abruptly to a diverse pico-nanoplanktonic community that represented ~95% of the total autotrophic C biomass. 'Chlorophytes', 'cryptophytes' and 'haptophytes II' in the nanoplankton, and 'chlorophytes' and 'Synechococcus' in the picoplankton were the main contributers to chl a in IPC waters, as calculated by CHEMTAX.

High-throughput sequencing (HTS) techniques have suggested the existence of a wealth of species with very low relative abundance: the rare biosphere. We attempted to exhaustively map this rare biosphere in two water samples by performing an exceptionally deep pyrosequencing analysis (~500,000 final reads per sample). Species data were derived by a 97% identity criterion and various parametric distributions were fitted to the observed counts. Using the best-fitting Sichel distribution we estimate a total species richness of 1,568–1,669 (95% Credible Interval) and 5,027–5,196 for surface and deep water samples respectively, implying that 84–89% of the total richness in those two samples was sequenced, and we predict that a quadrupling of the present sequencing effort would suffice to observe 90% of the total richness in both samples. Comparing the HTS results with a culturing approach we found that most of the cultured taxa were not obtained by HTS, despite the high sequencing effort. Culturing therefore remains a useful tool for uncovering marine bacterial diversity, in addition to its other uses for studying the ecology of marine bacteria.