Validation of the detection of Alexandrium species using specific RNA probes tested in a microarray format: Calibration of signal using variability of RNA content with environmental conditions.

The dinoflagellate genus Alexandrium contains several toxin producing species and strains, which can cause major economic losses to the shell fish industry. It is therefore important to be able to detect these toxin producers and also distinguish toxic strains from some of the morphologically identical non-toxic strains. To facilitate this DNA probes to be used in a microarray format were designed in silico or developed from existing published probes. These probes targeted either the 18S or 28S ribosomal ribonucleic acid (rRNA) gene in Alexandrium tamarense Group I, Group III and Group IV, Alexandrium ostenfeldii and Alexandrium minutum. Three strains of A. tamarense Group I, A. tamarense Group III, A. minutum and two strains of A. ostenfeldii were grown at optimal conditions and transferred into new environmental conditions changing either the light intensity, salinity, temperature or nutrient concentrations, to check if any of these environmental conditions induced changes in the cellular ribonucleic acid (RNA) concentration or growth rate. The aim of this experiment was the calibration of several species-specific probes for the quantification of the toxic Alexandrium strains. Growth rates were highly variable but only elevated or lowered salinity significantly lowered growth rate for A. tamarense Group I and Group III; differences in RNA content were not significant for the majority of the treatments. Only light intensity seemed to affect significantly the RNA content in A. tamarense Group I and Group III, but this was still within the same range as for the other treatments meaning that a back calibration from RNA to cell numbers was possible. The designed probes allow the production of quantitative information for Alexandrium species for the microarray chip.

Validation of the detection of Pseudo-nitzschia spp. using specific RNA probes tested in a microarray format: Calibration of signal based on variability of RNA content with environmental conditions.

Harmful algal blooms (HAB) occur worldwide and cause health problems and economic damage to fisheries and tourism. Monitoring for toxic algae is therefore essential but is based primarily on light microscopy, which is time consuming and can be limited by insufficient morphological characters such that more time is needed to examine critical features with electron microscopy. Monitoring with molecular tools is done in only a few places world-wide. EU FP7 MIDTAL (Microarray Detection of Toxic Algae) used SSU and LSU rRNA genes as targets on microarrays to identify toxic species. In order to comply with current monitoring requirements to report cell numbers as the relevant threshold measurement to trigger closure of fisheries, it was necessary to calibrate our microarray to convert the hybridisation signal obtained to cell numbers. Calibration curves for two species of Pseudo-nitzschia for use with the MIDTAL microarray are presented to obtain cell numbers following hybridisation. It complements work presented by Barra et al. (2012b. Environ. Sci. Pollut. Res. doi: 10.1007/s11356-012-1330-1v) for two other Pseudo-nitzschia spp., Dittami and Edvardsen (2012a. J. Phycol. 48, 1050) for Pseudochatonella, Blanco et al. (2013. Harmful Algae 24, 80) for Heterosigma, McCoy et al. (2013. FEMS. doi: 10.1111/1574-6941.12277) for Prymnesium spp., Karlodinium veneficum, and cf. Chatonella spp. and Taylor et al. (2014. Harmful Algae, in press) for Alexandrium.

The Partitioning Of Secondary Production Among Species In Benthic Communities

The way in which total secondary production is partitioned amongst species in various macrofauna communities (Amphiura, Venus, Abra, Modiolus) around the British Isles is discussed. When the proportion of total production is plotted for each species, ranked in order of productive importance, curves are produced which are characteristic of particular physical conditions. The shapes of the curves are independent of the actual species involved, but depend on the proportion of individuals in the community which adopt a particular feeding behaviour, and the scope for diversification within trophic groups. The form of these curves correlates closely with bottom currents and associated bed-stresses, since these affect both the nature of the food supply to bottom animals and the nature of the substrate. These observations have important implications for the structure and functioning of benthic communities. Comparison of production partitioning in the meiofauna of mud and sand substrates indicates a remarkable similarity within trophic groups although the partitioning of production between trophic groups is very different. The shapes of production-rank curves again appear to depend on the scope for diversification within trophic groups. In the meiofauna resources are partitioned more equitably than in the macrofauna. There is a marked discontinuity in the lognormal distribution of body sizes within integrated benthic communities at the meiofauna-macrofauna size boundary.

Distribution Of Sublittoral Macrofauna Communities In Bristol Channel In Relation To Substrate

Grab and dredge samples have been collected on a grid of 155 sublittoral stations in the Bristol Channel. The faunal data have been analysed using a hierarchical sorting technique to cluster stations with similar species compositions. At a similarity level of 18%, groups of stations with a species composition similar to the classical Petersen communities were defined. Three of Petersen's communities were recognized in the outer part of the Channel, the Venus, Abra and Modiolus communities. The fauna of the inner part of the Channel is reduced and does not correspond with any previously recognized community type. Possible causes for this faunal reduction are discussed. The substrate distribution and the macrofaunal community distribution are mapped. Side-scan sonograms are shown to be a useful adjunct to the interpretation of faunal distributions.

Identification of a diagnostic marker to detect freshwater cyanophages of filamentous cyanobacteria

Cyanophages are viruses that infect the cyanobacteria, globally important photosynthetic microorganisms. Cyanophages are considered significant components of microbial communities, playing major roles in influencing host community diversity and primary productivity, terminating cyanobacterial water blooms, and influencing biogeochemical cycles. Cyanophages are ubiquitous in both marine and freshwater systems; however, the majority of molecular research has been biased toward the study of marine cyanophages. In this study, a diagnostic probe was developed to detect freshwater cyanophages in natural waters. Oligonucleotide PCR-based primers were designed to specifically amplify the major capsid protein gene from previously characterized freshwater cyanomyoviruses that are infectious to the filamentous, nitrogen-fixing cyanobacterial genera Anabaena and Nostoc. The primers were also successful in yielding PCR products from mixed virus communities concentrated from water samples collected from freshwater lakes in the United Kingdom. The probes are thought to provide a useful tool for the investigation of cyanophage diversity in freshwater environments.