Electrochemical detection of the toxic dinoflagellate Alexandrium ostenfeldii with a DNA-biosensor

The steady rise of observations of harmful or toxic algal blooms throughout the world in the past decades constitute a menace for coastal ecosystems and human interests. As a consequence, a number of programs have been launched to monitor the occurrence of harmful and toxic algae. However, the identification is currently done by microscopic examination, which requires a broad taxonomic knowledge, expensive equipment and is very time consuming. In order to facilitate the identification of toxic algae, an inexpensive and easy-to-handle DNA-biosensor has been adapted for the electrochemical detection of the toxic dinoflagellate Alexandrium ostenfeldii. The detection of the toxic algae is based on a sandwich hybridisation, which is carried out on a disposable sensor chip. A set of two probes for the species specific identification of A. ostenfeldii was developed. The specificity of the probes could be shown in dot-blot hybridisation and with the DNA-biosensor. The sensitivity of the DNA-biosensor was optimised with respect to hybridisation temperature and NaCl-concentration and a significant increase of the sensitivity of the DNA-biosensor could be obtained by a fragmentation of the rRNA prior to the hybridisation and by adding a helper oligonucleotide, which binds in close proximity to the probes to the hybridisation

Molecular sensor-based monitoring of toxic algae

Planktonic algae are the most abundant photosynthetic organisms. They are the basis of the marine food web, and changes in phytoplankton communities generally provide an early indication for climate-driven modifications of marine food webs and the whole ecosystem. Harmful algal blooms (HAB) are a subset of planktonic algal species that have negative impacts on humans and aquatic environments. The formation of HABs depends on several multifactorial climate and environmental drivers that influence timing and frequency of these algal blooms. There is some evidence that the occurrence of HAB might become more intense, widespread, frequent and unexpected in future decades due to climate variability. In North ocean regions the global climate change is reflected by changing environmental parameter, such as increasing sea surface temperature, modified water mass stratification and rising freshwater content in the Arctic realm. Within this study the occurrence, abundance and dispersal of toxic algae species is analysed in Nordic seas and the Arctic Ocean, based on molecular detection with a nucleic acid biosensor. Alongside with this it will be assessed whether biosensor systems can serve as an early warning system for human health policies in the field of water resource management

Insight into protist diversity in Arctic sea ice and melt-pond aggregate obtained by pyrosequencing

Protists in the central Arctic Ocean are adapted to the harsh environmental conditions of its various habitats. During the Polarstern cruise ARK-XXVI/3 in 2011, at one sea-ice station, large aggregates accumulated at the bottom of the melt ponds. In this study, the protist assemblages of the bottom layer of the sea-ice and melt-pond aggregate were investigated using flow cytometry and 454-pyrosequencing. The objective is to provide a first molecular overview of protist biodiversity in these habitats and to consider the overlaps and/or differences in the community compositions. Results of flow cytometry pointed to a cell size distribution that was dominated by 3–10 µm nanoflagellates. The phylogenetic classification of all sequences was conducted at a high taxonomic level, while a selection of abundant (≥1% of total reads) sequences was further classified at a lower level. At a high taxonomic level, both habitats showed very similar community structures, dominated by chrysophytes and chlorophytes. At a lower taxonomic level, dissimilarities in the diversity of both groups were encountered in the abundant biosphere. While sea-ice chlorophytes and chrysophytes were dominated by Chlamydomonas/Chloromonas spp. and Ochromonas spp., the melt-pond aggregate was dominated by Carteria sp., Ochromonas spp. and Dinobryon faculiferum. We suppose that the similarities in richness and community structure are a consequence of melt-pond freshwater seeping through porous sea ice in late summer. Differences in the abundant biosphere nevertheless indicate that environmental conditions in both habitats vary enough to select for different dominant species

Limited sinking of Phaeocystis during a 12 days sediment trap study

There is a controversy discussion about the contribution of the genus Phaeocystis to the vertical carbon export with evidence for and against sedimentation of Phaeocystis. So far, the presence of Phaeocystis in sinking matter was investigated with methods depending on morphological features (microscopy) and fast degradable substances (biochemical analyses). In this study, we determine the occurrence and abundance of Phaeocystis antarctica in short-term sediment traps and the overlying water column during a 12-day time period in the Atlantic sector of the Southern Ocean with 454-pyrosequencing and microscopy counting. In the sediment trap samples, we only found few sequences belonging to Phaeocystis, which was not reflecting the situation in the water column above. The cell counts showed the same results. We conclude that Phaeocystis cells are not generally transported downwards by active sinking or other sinking processes

Molecular analyses of gut contents: elucidating the feeding of co-occurring salps in the Lazarev Sea from a different perspective

The diet of Antarctic salps was elucidated by investigating their gut content using “Automated Ribosomal Intergenic Spacer Analysis” and 454-pyrosequencing. Salp samples were collected during the Lazarev Krill Study in the Western Weddell Sea (summer 2005/06 and 2007/08, fall 2004, and winter 2006). Two salp species, Salpa thompsoni and Ihlea racovitzai, both occur in the Southern Ocean, can overlap geographically and seasonally. Here we provide evidence that despite the non-selective feeding mechanism, the two co-occuring salp species might have different niches within one given habitat. ARISA-patterns of 93 salp gut content samples revealed strong differences between the two salp species, even at the same sampling site. These differences were confirmed by 454-pyrosequencing of the V4-18S rDNA of ten salps. The pyrosequencing data indicate that flagellates, in particular dinophyceae constitute a high proportion of the sequence reads identified in the gut content of both salp species. However, within the dinophyceae differences in the read composition were detected between the two salp species. This supports the findings of a previous study where fatty acid signatures indicate a flagellate based diet of salps, even though microscopic analyses identified diatoms as the dominant component of salp gut contents

Uncovering hidden biodiversity in the Cryptophyta: Clone library studies at the Helgoland Time Series Site in the Southern German Bight indentifies the cryptophycean clade potentially responsible for the majority of its genetic diversity during the spring bloom.

Cryptophyceae are important group in marine phytoplankton, but little is known about the occurrence and distribution of individual species. Recently, with use of molecular probes and microarray technology, it has been shown that species related to teleaulax spp. or Chroomonas spp. (clades 4 or 6) contributed the most to cryptophycean biomass in the north Sea. for the microarray study, the single probe (clade 4/6) recognizes members of both clades 4 and 6 and thus cannot separate them. Therefore, it was unknown as to whether the cryptophyte community was composed of clade 4, clade 6 or both of them. Here, we addressed this question and increased the genetic diversity of our investigations of cryptophycean diversity in the north Sea by sequencing 18S rRnA clone libraries made from fractionated water samples to examine specifically the picoplanktonic fraction because that fraction was studied in detail in the earlier microarray study. We focused on samples from the spring phytoplankton bloom in 2004 because the microarray signals were the strongest at this time. Excluding chimeric sequences, we detect- ed nine cryptophycean oTUs, seven of which fell into the teleaulax/Plagioselmis branch, whereas two grouped with Geminigera spp. our results indicate that these oTUs, affiliated with clade 4, may be an important component of cryptophyte community during spring bloom in the north Se