Inhibitory effects of Skeletonema costatum on mercury methylation by Geobacter sulfurreducens PCA.

Algae and mercury (Hg) are ubiquitous in marine environments. In this study, we investigated the effects of a typical marine algae of diatom Skeletonema costatum on Hg methylation by an iron-reducing bacterium of Geobacter sulfurreducens (G. sulfurreducens) PCA. In the absence of Skeletonema costatum, the bacterial MeHg production rate maximized at 104.06 ±â€¯11.7 ng L-1 h-1 with a high Hg level, while the highest methylation efficiency was achieved at a low Hg concentration. The existence of Skeletonema costatum greatly inhibited the capability of G. sulfurreducens PCA to methylate Hg. With the increase in algal biomass, there was a significant mitigation of MeHg formation and Hg0 release, leaving a considerable proportion of immobilized Hg2+ species (up to 47%) associated with algal cell materials. These results suggest that marine algae are crucial in determining the bioavailability of Hg contaminants and the methylating potential of G. sulfurreducens PCA.

A high resolution melting method for the molecular identification of the potentially toxic diatom Pseudo-nitzschia spp. in the Mediterranean Sea.

The aim of this study was to develop and validate a high resolution melting (HRM) method for the rapid, accurate identification of the various harmful diatom Pseudo-nitzschia species in marine environments. Pseudo-nitzschia has a worldwide distribution and some species are toxic, producing the potent domoic acid toxin, which poses a threat to both human and animal health. Hence, it is important to identify toxic Pseudo-nitzschia species. A pair of primers targeting the LSU rDNA of the genus Pseudo-nitzschia was designed for the development of the assay and its specificity was validated using 22 control DNAs of the P. calliantha, P. delicatissima/P. arenysensis complex and P. pungens. The post-PCR HRM assay was applied to numerous unidentified Pseudo-nitzschia strains isolated from the northwestern Adriatic Sea (Mediterranean Sea), and it was able to detect and discriminate three distinct Pseudo-nitzschia taxa from unidentified samples. Moreover, the species-specific identification of Pseudo-nitzschia isolates by the HRM assay was consistent with phylogenetic analyses. The HRM assay was specific, robust and rapid when applied to high numbers of cultured samples in order to taxonomically identify Pseudo-nitzschia isolates recovered from environmental samples.

Chitinase producing bacteria with direct algicidal activity on marine diatoms.

Chitinase producing bacteria can involve extensively in nutrient cycling and energy flow in the aquatic environment through degradation and utilization of chitin. It is well known that diatoms cells are encased by box-like frustules composed of chitin. Thus the chitin containing of diatoms shall be a natural target of chitinase producing bacteria, however, the interaction between these two organismic groups has not been studied thus far. Therefore, in this study, the algicidal mechanism of one chitinase producing bacterium (strain LY03) on Thalassiosira pseudonana was investigated. The algicidal range and algicidal mode of strain LY03 were first studied, and then bacterial viability, chemotactic ability and direct interaction characteristic between bacteria and diatom were also confirmed. Finally, the characteristic of the intracellular algicidal substance was identified and the algicidal mechanism was determined whereby algicidal bacterial cells showed chemotaxis to algal cells, fastened themselves on algal cells with their flagella, and then produced chitinase to degrade algal cell walls, and eventually caused algal lysis and death. It is the first time to investigate the interaction between chitinase producing bacteria and diatoms, and this novel special interaction mode was confirmed in this study, which will be helpful in protection and utilization of diatoms resources.

Iron enrichment stimulates toxic diatom production in high-nitrate, low-chlorophyll areas.

Oceanic high-nitrate, low-chlorophyll environments have been highlighted for potential large-scale iron fertilizations to help mitigate global climate change. Controversy surrounds these initiatives, both in the degree of carbon removal and magnitude of ecosystem impacts. Previous open ocean enrichment experiments have shown that iron additions stimulate growth of the toxigenic diatom genus Pseudonitzschia. Most Pseudonitzschia species in coastal waters produce the neurotoxin domoic acid (DA), with their blooms causing detrimental marine ecosystem impacts, but oceanic Pseudonitzschia species are considered nontoxic. Here we demonstrate that the sparse oceanic Pseudonitzschia community at the high-nitrate, low-chlorophyll Ocean Station PAPA (50 degrees N, 145 degrees W) produces approximately 200 pg DA L(-1) in response to iron addition, that DA alters phytoplankton community structure to benefit Pseudonitzschia, and that oceanic cell isolates are toxic. Given the negative effects of DA in coastal food webs, these findings raise serious concern over the net benefit and sustainability of large-scale iron fertilizations.

Toxigenic effects of diatoms on grazers, phytoplankton and other microbes: a review.

Traditionally, diatoms have been regarded as providing the bulk of the food that sustains the marine food chain and important fisheries. However, this view was challenged almost two decades ago on the basis of laboratory and field studies showing that when copepods, the principal predators of diatoms, feed on certain diatom diets, they produce abnormal eggs that either fail to develop to hatching or hatch into malformed (i.e. teratogenic) nauplii that die soon afterwards. Over the years, many explanations have been advanced to explain the causes for reproductive failure in copepods and other marine and freshwater invertebrates including diatom toxicity, or nutritional deficiency and poor assimilation of essential compounds in the animal gut. Here we review the literature concerning the first possibility, that diatoms produce cytotoxic compounds responsible for growth inhibition and teratogenic activity, potentially sabotaging future generations of grazers by inducing poor recruitment. The cytotoxic compounds responsible for these effects are short chain polyunsaturated aldehydes (PUAs) and other oxygenated fatty acid degradation products such as hydroxides, oxo-acids, and epoxyalcohols (collectively termed oxylipins) that are cleaved from fatty acid precursors by enzymes activated within seconds after crushing of cells. Such toxins are suggested to have multiple simultaneous functions in that they not only deter herbivore feeding but some also act as allelopathic agents against other phytoplankton cells, thereby affecting the growth of competitors, and also signalling population-level cell death and termination of blooms, with possible consequences for food web structure and community composition. Some oxylipins also play a role in driving marine bacterial community diversity, with neutral, positive or negative interactions depending on the species, thereby shaping the structure of bacterial communities during diatom blooms. Several reviews have already been published on diatom-grazer interactions so this paper does not attempt to provide a comprehensive overview, but rather to consider some of the more recent findings in this field. We also consider the role of diatom oxylipins in mediating physiological and ecological processes in the plankton and the multiple simultaneous functions of these secondary metabolites.

Biocidal performance of acrylated glyphosate in a model photopolymerizable coating formulation.

Acrylated glyphosate was blended into a model polyolacrylate formulation and copolymerized. The resulting copolymer retains herbicidal activity similar to that of the monomer as indicated by the results of biological tests. No release of biocide from the coating was observed. The potential value of these biologically active acrylic formulations as biofouling compositions has been demonstrated by field trials.

Proliferaciones algales de la diatomea toxigénica Pseudo-Nitzschia (Bacillariophyceae) en el Golfo de Nicoya, Costa Rica / Algal blooms of the toxigenic diatom Pseudo-Nitzschia (Bacillariophyceae) in the Golfo de Nicoya, Costa Rica

En el mes de noviembre de 2001, se aisló de una marea roja cerca de la Isla San Lucas, las diatomeas Pseudo-Nitzschia pungens f. pungens seguida por Skeletonema costatum, Chaetoceros lorenzianus y en menor concentración Thalassiosira spp. las cuales fueron caracterizadas por microscopía electrónica de rastreo y transmisión, siendo esta la primera vez que se describe la presencia de estas especies produciendo proliferaciones en Costa Rica. Actualmente existe un aumento en el reporte de mareas rojas donde predominan las diatomeas, y los conteos celulares indican el aumento en el número de las especies conocidas como Pseudo-nitzschia pungens f. multiseries. A estas se les atribuye la producción del ácido domóico, un aminoácido de bajo peso molecular. Las intoxicaciones amnésicas por mariscos (IAM) que afectan a los humanos, se producen por la ingesta de mariscos contaminados con este ácido. En Costa Rica, hasta la fecha, solo se han reportado casos de intoxicación paralítica por mariscos contaminados por los dinoflagelados Pyrodinium bahamense var. compressum y Gymnodinium catenatum, sin embargo existe la posibilidad de producirse intoxicaciones humanas por la ingesta de mariscos y peces contaminados con toxinas amnésicas las cuales son hidrosolubles y termoestables. Debe entonces considerarse el peligro potencial de aparición de casos de intoxicaciones amnésicas y se sugiere la inclusión de estas especies en los programas de monitoreo permanente para tomar las medidas preventivas de salud pública

Water samples were collected during a red tide event in November 2001, near San Lucas Island (Gulf of Nicoya, Costa Rica). Superficial temperature was 27ºC and water was turbid, with no fetid smell. One sample was treated with negative staining and observed using a transmission electron microscope (TEM); another sample was observed using a scanning electron microscope (SEM). Samples had high concentrations of the diatom Pseudo-Nitzschia pungens f. pungens (characterized by two rows of poroids in the external channel), and lower concentrations of Skeletonema costatum (chains joined by external microtubules) and Chaetoceros lorenzianus (oval apertures and long chains, having setae with distinctive transverse rows and spines). This is the first time that the first species was described producing red tides in Costa Rica. However, reports about red tides with high concentration of species like P. pungens (variety multiseries) are increasing. These species have been related to the production of domoic acid, a low molecular weight amino acid which in humans can cause amnesic intoxications with seafood. Previously, Costa Rican reports of toxic accidents only referred to seafood contaminated with Pyrodinium bahamense var. compressum and Gymnodinium catenatum dinoflagellates. The increase in the number of Pseudo-Nitzschia causing harmful algae blooms is of interest for scientists around the world and must be documented. Similarly, some Chaetoceros species have been reported to be harmful to fish. We strongly recommend the establishment of a permanent surveillance program monitoring the presence of these species new at Costa Rican Pacific coast. Since the amnesic toxin is soluble in water and heat-resistant, we want to stress the possibility of having human cases of amnesic intoxication

Domoic acid accumulation in French shellfish in relation to toxic species of Pseudo-nitzschia multiseries and P. pseudodelicatissima.

Within the French phytoplankton monitoring network (REPHY), domoic acid (DA), the toxin responsible for amnesic shellfish poisoning, was first detected in samples collected in 1998. Toxin analysis by the official method [liquid chromatography with diode array detection (LC/DAD)] was performed when Pseudo-nitzschia cell concentration was greater than 1.0 x 10(5) cells/l. LC/DAD results obtained in 1999 and 2000 showed increased DA accumulation in bivalves sampled at different sites along French coasts. The toxin maximum in 1999 was 3.2 microg DA/g of whole tissue, whereas the levels in 2000 (53 microg) were above the sanitary threshold (20 microg DA/g tissue). Phytoplankton samples collected during blooms were observed by both light and scanning electron microscopy (SEM). Identification of phytoplankton species by SEM analyses confirmed the presence of two known DA-producing species, P. pseudodelicatissima and P. multiseries. LC/DAD results for a mass culture of P. multiseries indicated that this species was involved in DA accumulation in French shellfish.

Detection of stable pre-rRNA in toxigenic Pseudo-nitzschia species.

Nucleotide sequence analysis of ribosomal DNA (rDNA) spacer regions is useful for taxonomic comparisons of closely related microorganisms. These regions have been less useful for routine microbial identification and detection, partly because rRNA precursors (pre-rRNAs) in microbial cells are assumed to be too labile to be detectable by high-throughput probe hybridization methods. We characterized the sequence diversity and physiological stability of pre-rRNA in the toxigenic marine diatoms Pseudo-nitzschia australis, P. multiseries, and P. pungens. As with nucleotide sequences of the first internal transcribed spacer (ITS1) reported previously, sequences of ITS2 and the 5' external transcribed spacer (ETS1) exhibited considerable divergence among these species, including large insertions-deletions detectable by PCR-based spacer length analysis. In slot blot hybridization assays on RNA extracted from lysates of Pseudo-nitzschia cells, oligonucleotide probes directed to pre-rRNA spacers generated much stronger signals than did complementary probes directed to the coding strands of the rDNAs, indicating that the pre-rRNA-targeted probes detected multicopy transcripts. A group of probes directed to a discrete 90-base region within the ITS1 pre-rRNA gave no detectable signal, suggesting that this region is degraded early in the rRNA maturation pathway. Other pre-rRNA regions were always detectable and, in marked contrast to prokaryotic systems analyzed in this manner, were stable and abundant in both actively dividing and nondividing cells. Long, multilabeled RNA probes, which would exhibit considerable cross-reactivity if directed to mature rRNA sequences, detected species-specific pre-rRNA sequences from as few as 1,000 cells. Pre-rRNA is a potentially useful molecular target for detecting and identifying Pseudo-nitzschia species and possibly other unicellular eukaryotes as well.