Numerical modeling of microplastic interaction with fine sediment under estuarine conditions.

Microplastic (MP) pollution is an important challenge for human life which has consequently affected the natural system of other organisms. Mismanagement and also careless handling of plastics in daily life has led to an accelerating contamination of air, water and soil compartments with MP. Under estuarine conditions, interactions with suspended particulate matter (SPM) like fine sediment in the water column play an important role on the fate of MP. Further studies to better understand the corresponding transport and accumulation mechanisms are required. This paper aims at providing a new modeling approach improving the MP settling velocity formulation based on higher suspended fine sediment concentrations, as i.e. existent in estuarine turbidity zones (ETZ). The capability of the suggested approach is examined through the modeling of released MP transport in water and their interactions with fine sediment (cohesive sediment/fluid mud). The model results suggest higher concentrations of MP in ETZ, both in the water column as well as the bed sediment, which is also supported by measurements. The key process in the modeling approach is the integration of small MP particles into estuarine fine sediment aggregates. This is realized by means of a threshold sediment concentration, above which the effective MP settling velocity increasingly approaches that of the sediment aggregates. The model results are in good agreement with measured MP mass concentrations. Moreover, the model results also show that lighter small MP particles can easier escape the ETZ towards the open sea.

Low numbers of microplastics detected in drinking water from ground water sources.

Microplastic particles have been detected in various natural habitats and the digestive tracts of several species. These particles have also been reported in commercially available seafood, salt or bottled water starting discussions on potential implications for human health. To be able to assess the related risks, exposure concentrations and pathways need to be known. Here, we analysed ground water and drinking water for the presence of microplastics (>20 µm) using FTIR imaging. Samples were taken at different positions within the drinking water supply chain. Determined concentrations ranged from 0 to 7 microplastics m-3 raw water or drinking water with an overall mean of 0.7 microplastics m-3. These particles were identified as polyethylene, polyamide, polyester, polyvinylchloride or epoxy resin and between 50 and 150 µm in size. Plastic is a resistant and durable material, still, the abrasion of plastic equipment used during water purification or transport is a likely explanation for the plastic particles detected in water samples.

Identification of microplastic in effluents of waste water treatment plants using focal plane array-based micro-Fourier-transform infrared imaging.

The global presence of microplastic (MP) in aquatic ecosystems has been shown by various studies. However, neither MP concentrations nor their sources or sinks are completely known. Waste water treatment plants (WWTPs) are considered as significant point sources discharging MP to the environment. This study investigated MP in the effluents of 12 WWTPs in Lower Saxony, Germany. Samples were purified by a plastic-preserving enzymatic-oxidative procedure and subsequent density separation using a zinc chloride solution. For analysis, attenuated total reflection Fourier-transform infrared spectroscopy (ATR-FT-IR) and focal plane array (FPA)-based transmission micro-FT-IR imaging were applied. This allowed the identification of polymers of all MP down to a size of 20 µm. In all effluents MP was found with quantities ranging from 0 to 5 × 101 m-3 MP > 500 µm and 1 × 101 to 9 × 103 m-3 MP < 500 µm. By far, polyethylene was the most frequent polymer type in both size classes. Quantities of synthetic fibres ranged from 9 × 101 to 1 × 103 m-3 and were predominantly made of polyester. Considering the annual effluxes of tested WWTPs, total discharges of 9 × 107 to 4 × 109 MP particles and fibres per WWTP could be expected. Interestingly, one tertiary WWTP had an additionally installed post-filtration that reduced the total MP discharge by 97%. Furthermore, the sewage sludge of six WWTPs was examined and the existence of MP, predominantly polyethylene, revealed. Our findings suggest that WWTPs could be a sink but also a source of MP and thus can be considered to play an important role for environmental MP pollution.

Comparison of molecular species identification for North Sea calanoid copepods (Crustacea) using proteome fingerprints and DNA sequences.

Calanoid copepods play an important role in the pelagic ecosystem making them subject to various taxonomic and ecological studies, as well as indicators for detecting changes in the marine habitat. For all these investigations, valid identification, mainly of sibling and cryptic species as well as early life history stages, represents a central issue. In this study, we compare species identification methods for pelagic calanoid copepod species from the North Sea and adjacent regions in a total of 333 specimens. Morphologically identified specimens were analysed on the basis of nucleotide sequences (i.e. partial mitochondrial cytochrome c oxidase subunit I (COI) and complete 18S rDNA) and on proteome fingerprints using the technology of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). On all three molecular approaches, all specimens were classified to species level indicated by low intraspecific and high interspecific variability. Sequence divergences in both markers revealed a second Pseudocalanus species for the southern North Sea identified as Pseudocalanus moultoni by COI sequence comparisons to GenBank. Proteome fingerprints were valid for species clusters irrespective of high intraspecific variability, including significant differences between early developmental stages and adults. There was no effect of sampling region or time; thus, trophic effect, when analysing the whole organisms, was observed in species-specific protein mass spectra, underlining the power of this tool in the application on metazoan species identification. Because of less sample preparation steps, we recommend proteomic fingerprinting using the MALDI-TOF MS as an alternative or supplementary approach for rapid, cost-effective species identification.

[Vibrio infections from food and sea water. Introducing the "VibrioNet"]. / Vibrio-Infektionen durch Lebensmittel und Meerwasser. Das Netzwerk "VibrioNet" stellt sich vor.

Vibrio is a genus of bacteria present in surface and coastal waters as well as in marine organisms worldwide. In many countries, pathogenic Vibrio species are a main cause of bacterial diarrhea, which may result from comsumption of contaminated seafood and fish products or from drinking contaminated water. Vibrio infections may also gain in importance in our regions due to global warming and the increase in the world trade of seafood. The research network "VibrioNet" studies pathogenic Vibrios in the marine environment and in seafood consumed by humans as a potential, new emerging zoonotic agent. An assessment of the risk arising from pathogenic non-cholera-vibrios in central Europe is the target of a multidisciplinary research effort. The research network will be strengthened by cooperations with international partners from countries in which Vibrio infections play a major role (Bangladesh, Chile, India, Thailand, and Vietnam).

Populations of heavy fuel oil-degrading marine microbial community in presence of oil sorbent materials.

AIMS: To investigate the feasibility of applying sorbent material X-Oil in marine oil spill mitigation and to survey the interactions of oil, bacteria and sorbent. METHODS AND RESULTS: In a series of microcosms, 25 different treatments including nutrient amendment, bioaugmentation with Alcanivorax borkumensis and application of sorbent were tested. Microbial community dynamics were analysed by DNA fingerprinting methods, RISA and DGGE. Results of this study showed that the microbial communities in microcosms with highly active biodegradation were strongly selected in favour of A. borkumensis. Oxygen consumption measurements in microcosms and gas chromatography of oil samples indicated the fast and intense depletion of linear alkanes as well as high oxygen consumption within 1 week followed by consequent slower degradation of branched and polyaromatic hydrocarbons. CONCLUSION: Under given conditions, A. borkumensis was an essential organism for biodegradation, dominating the biofilm microbial community formation and was the reason of emulsification. SIGNIFICANCE AND IMPACT OF THE STUDY: This study strongly emphasizes the pivotal importance of A. borkumensis as an essential organism in the initial steps of marine hydrocarbon degradation. Interaction with the sorbent material X-Oil proved to be neutral to beneficial for biodegradation and also promoted the growth of yet unknown micro-organisms.

FISH and chips: marine bacterial communities analyzed by flow cytometry based on microfluidics.

To unveil the structure of natural marine pelagic bacterial communities, PCR-based techniques as well as fluorescence in situ hybridizations (FISH) were successfully performed in the past. Using fluorescence microscopes or confocal laser scanning microscopes (CLSM) for the analysis of FISH experiments, it was possible to differentiate bacterial communities, but most attempts to combine flow cytometry and FISH for this purpose have failed till now. Here we present a successful analysis of FISH experiments of natural marine pelagic bacterial communities using a flow cytometer based on microfluidics (Agilent 2100 bioanalyzer). Marine water samples were enriched on polycarbonate filters and hybridized with Cy5 labeled gene probes of different phylogenetic depth. Bacteria were detached from the filters and subsequently analyzed in the Cell Chip of the Agilent 2100 Bioanalyzer. Samples were counter-stained using SYTOX. In all samples the EUB338 positive signals could be clearly differentiated from those of the NON probe. Furthermore a dominance of alpha-protebacteria (as indicated by the probes ALF968 and G rB) could be observed. Microfluidics based flow cytometry is a promising technique for the analysis of natural bacterial communities from the marine environment.

Overview of key phytoplankton toxins and their recent occurrence in the North and Baltic Seas.

The frequency and intensity of harmful algal blooms (HABs) appear to be on the rise globally. There is also evidence of the geographic spreading of toxic strains of these algae. Consequently, methods had to be established and new ones are still needed for the evaluation of possible hazards caused by increased algal toxin production in the marine food chain. Different clinical effects of algae-related poisoning have attracted scientific attention; paralytic shellfish poisoning, diarrhetic shellfish poisoning, and amnesic shellfish poisoning are among the most common. Additionally, cyanobacteria (blue-green algae) in brackish waters often produce neurotoxic and hepatotoxic substances. Bioassays with mice or rats are common methods to determine algal and cyanobacterial toxins. However, biological tests are not really satisfactory because of their low sensitivity. In addition, there is growing public opposition to animal testing. Therefore, there has been increasing effort to determine algal toxins by chemical methods. Plankton samples from different European marine and brackish waters were taken during research cruises and analyzed on board directly. The ship routes covered marine areas in the northwest Atlantic, Orkney Islands, east coast of Scotland, and the North and Baltic seas. The first results on the occurrence and frequency of harmful algal species were obtained in 1997 and 1998. During the 2000 cruise an HPLC/MS coupling was established on board, and algal toxins were measured directly after extraction of the plankton samples. In contrast to earlier cruises, the sampling areas were changed in 2000 to focusing on coastal zones. The occurrence of toxic algae in these areas was compared to toxin formation during HABs in the open sea. It was found that the toxicity of the algal blooms depended on the prevailing local conditions. This observation was also confirmed by monitoring cyanobacterial blooms in the Baltic Sea. Optimal weather conditions, for example, during the summers of 1997 and 2003, favored blooms of cyanobacteria in all regions of the Baltic. The dominant species regarding the HABs in the Baltic was Nodularia spumigena. However, in addition to high concentrations of Nodularia spumigena in coastal zones, other blue-green algae are involved in bloom formation, with changes in plankton communities influencing both toxin profiles and toxicity.

Isolation of novel pelagic bacteria from the German bight and their seasonal contributions to surface picoplankton.

We tested new strategies for the isolation of abundant bacteria from coastal North Sea surface waters, which included reducing by several orders of magnitude the concentrations of inorganic N and P compounds in a synthetic seawater medium. Agar plates were resampled over 37 days, and slowly growing colonies were allowed to develop by repeatedly removing all newly formed colonies. A fivefold increase of colonies was observed on plates with reduced nutrient levels, and the phylogenetic composition of the culture collection changed over time, towards members of the Roseobacter lineage and other alpha-proteobacteria. Novel gamma-proteobacteria from a previously uncultured but cosmopolitan lineage (NOR5) formed colonies only after 12 days of plate incubation. A time series of German Bight surface waters (January to December 1998) was screened by fluorescence in situ hybridization (FISH) with isolate-specific and general probes. During spring and early summer, a prominent fraction of FISH-detectable bacteria (mean, 51%) were affiliated with the Cytophaga-Flavobacterium group (CF) of the Bacteroidetes. One Cytophaga sp. lineage with cultured representatives formed almost 20% of the CF group. Members of the Roseobacter cluster constituted approximately 50% of alpha-proteobacteria, but none of the Roseobacter-related isolates formed populations of >1% in the environment. Thus, the readily culturable members of this clade are probably not representative of Roseobacter species that are common in the water column. In contrast, members of NOR5 were found at high abundances (>10(5) cells ml(-1)) in the summer plankton. Some abundant pelagic bacteria are apparently able to form colonies on solid media, but appropriate isolation techniques for different species need to be developed.

Neuroactive compounds produced by bacteria from the marine sponge Halichondria panicea: activation of the neuronal NMDA receptor.

Previous studies revealed that the marine sponge Halichondria panicea habors symbiotic- and commensalic bacteria (Althoff et al., 1998. Marine Biol. 130, 529-536). In the present study the hypothesis was tested whether some of those bacteria synthesize neuroactive compounds. For the first time the effect of bacterial bioactive compounds on the neuronal ionotropic glutamate receptors [iGluR], subtype N-methyl-d-aspartate (NMDA) receptor, was checked. In cortical neurons from rats as cell system the supernatant of two bacterial cultures isolated from H. panicea proved to agonize the NMDA receptor. The response of the NMDA receptor to the bioactive compounds was determined by measuring the intracellular Ca(2+) level. The supernatants of cultures 697 and 698 were found to upregulate the intracellular Ca(2+) level. To validate the specificity of the effects, inhibition studies with Memantine and d-AP5 were performed. The two bacteria were identified by polymerase chain reaction-amplification of the 16S rDNA genes and subsequent sequencing; they displayed highest identity to Antarcticum vesiculatum and to Psychroserpens burtonensis, respectively. Based on these data first experimental evidence is presented indicating that bacteria associated with sponges display neuroactivity by agonizing the NMDA receptor.

Bacteria of the Genus Roseobacter Associated with the Toxic Dinoflagellate Prorocentrum lima.

The dinoflagellate Prorocentrum lima is known to produce diarrhetic shellfish poisons. However, it is yet unclear if the dinoflagellates themselves or the bacteria associated with them produce the toxins. Here we analyze the toxicity as well as the spectrum of bacteria in two cultures of P. lima, namely P. lima-SY and P. lima-ST, which initially derived from the same P. lima strain PL2V. Toxicity tests, applying the Artemia bioassay revealed in both cultures high levels of toxins. The bacteria, associated with the two cultures, were identified by PCR/nucleotide sequence analysis of the 16S rRNA gene. From cultures of P. lima-SY the dominant sequence was found to share a 93.7% similarity with the sequence of Roseobacter algocolus [R. algicola]; the relative abundance was determined to be 83%. In addition three further sequences of bacteria, grouped to the α-Protobacteria have been identified: Paracoccus denitrificans [90.8%], R. algocolus [94.4%] and Rhizobium huakuii [92.6%]. The identification of bacteria in P. lima-ST revealed that most share highest similarity with Bartonella taylorii but with a relatively low score of 87%. In addition to this sequence, two sequences with high similarity to the genus Roseobacter were obtained. The other sequences identified have not been detected in P. lima-SY. Studies with pure bacterial strains, previously isolated from a culture of P. lima-ST and subsequently cultured on agar plates, revealed that none of them was identical to those identified in the dinoflagellate culture itself. An explanation for the change of the spectrum of bacteria in the different cultures can only be expected when axenic cultures from P. lima are available.