Temporal variability of the microbial food web (viruses to ciliates) under the influence of the Black Sea Water inflow (N. Aegean, E. Mediterranean)

Τhe entire pelagic microbial food web was studied during the winter-spring period in the frontal area of the North Aegean Sea. Abundance of viruses, heterotrophic bacteria, cyanobacteria, auto- and hetero-trophic flagellates, and ciliates, as well as bacterial production, were measured at three stations (MD1, MD2, MD3) situated along a N-S transect between the area directly influenced by the inflowing Black Sea water and the area covered by the Levantine water. Samples were collected in December 2009, and January, March, April, and May 2011. Station MD1 exhibited the highest values of abundance and integrated biomass of all microbial groups and bacterial production during all months, and MD3 the lowest. Bacteria dominated the total integrated biomass at all stations and months, followed by cyanobacteria, auto-, hetero-trophic flagellates and ciliates. On a temporal scale, the microbial food web was less important in March as all microbial parameters at all stations showed the lowest values. After the phytoplankton bloom in March, the heterotrophic part of the microbial food web (mainly) strongly increased, though the intensity of the phenomenon was diminished from North to South. Pico-sized plankton was found to be heterotrophic whereas nanoplankton was autotrophic. It seems that the influence of the Black Sea water on station MD1, permanent throughout the study period of early winter to late spring, was reflected in all microbial populations studied, and produced a more productive pelagic food web system, with potential consequences for the upper trophic levels

Temporal variability of the microbial food web (viruses to ciliates) under the influence of the Black Sea Water inflow (N. Aegean, E. Mediterranean)

Τhe entire pelagic microbial food web was studied during the winter-spring period in the frontal area of the North Aegean Sea. Abundance of viruses, heterotrophic bacteria, cyanobacteria, auto- and hetero-trophic flagellates, and ciliates, as well as bacterial production, were measured at three stations (MD1, MD2, MD3) situated along a N-S transect between the area directly influenced by the inflowing Black Sea water and the area covered by the Levantine water. Samples were collected in December 2009, and January, March, April, and May 2011. Station MD1 exhibited the highest values of abundance and integrated biomass of all microbial groups and bacterial production during all months, and MD3 the lowest. Bacteria dominated the total integrated biomass at all stations and months, followed by cyanobacteria, auto-, hetero-trophic flagellates and ciliates. On a temporal scale, the microbial food web was less important in March as all microbial parameters at all stations showed the lowest values. After the phytoplankton bloom in March, the heterotrophic part of the microbial food web (mainly) strongly increased, though the intensity of the phenomenon was diminished from North to South. Pico-sized plankton was found to be heterotrophic whereas nanoplankton was autotrophic. It seems that the influence of the Black Sea water on station MD1, permanent throughout the study period of early winter to late spring, was reflected in all microbial populations studied, and produced a more productive pelagic food web system, with potential consequences for the upper trophic levels

Spatial variability in Antarctic surface snow bacterial communities

It was once a long-held view that the Antarctic was a pristine environment with low biomass, low biodiversity and low rates of microbial activity. However, as the intensity of scientific investigation has increased, so these views have started to change. In particular, the role and impact of human activity toward indigenous microbial communities has started to come under more intense scrutiny. During the Subglacial Lake Ellsworth exploration campaign in December 2012, a microbiological survey was conducted to determine the extent and likelihood of exogenous input into the subglacial lake system during the hot-water drilling process. Snow was collected from the surface to represent that used for melt water production for hot-water drilling. The results of this study showed that snow used to provide melt water differed in its microbiological composition from that of the surrounding area and raised the question of how the biogeography of snow-borne microorganisms might influence the potential outcome of scientific analyses. In this study, we investigated the biogeography of microorganisms in snow around a series of Antarctic logistic hubs, where human activity was clearly apparent, and from which scientific investigations have been undertaken. A change in microbial community structure with geographical location was apparent and, notably, a decrease in alpha diversity at more remote southern latitudes. Soil-related microorganisms dominated microbial assemblages suggesting terrestrial input, most likely from long-range aeolian transport into continental Antarctica. We also observed that relic DNA was not a major issue when assessing snow samples. Overall, our observations might have profound implications for future scientific activities in Antarctica, such as the need to establish “no-go” protected areas, the need for better characterization of field sites and improved protocols for sterilization and verification of ice drilling equipment

Microbiology: lessons from a first attempt at Lake Ellsworth

During the attempt to directly access, measure and sample Subglacial Lake Ellsworth in 2012–2013, we conducted microbiological analyses of the drilling equipment, scientific instrumentation, field camp and natural surroundings. From these studies, a number of lessons can be learned about the cleanliness of deep Antarctic subglacial lake access leading to, in particular, knowledge of the limitations of some of the most basic relevant microbiological principles. Here, we focus on five of the core challenges faced and describe how cleanliness and sterilization were implemented in the field. In the light of our field experiences, we consider how effective these actions were, and what can be learnt for future subglacial exploration missions. The five areas covered are: (i) field camp environment and activities, (ii) the engineering processes surrounding the hot water drilling, (iii) sample handling, including recovery, stability and preservation, (iv) clean access methodologies and removal of sample material, and (v) the biodiversity and distribution of bacteria around the Antarctic. Comparisons are made between the microbiology of the Lake Ellsworth field site and other Antarctic systems, including the lakes on Signy Island, and on the Antarctic Peninsula at Lake Hodgson. Ongoing research to better define and characterize the behaviour of natural and introduced microbial populations in response to deep-ice drilling is also discussed. We recommend that future access programmes: (i) assess each specific local environment in enhanced detail due to the potential for local contamination, (ii) consider the sterility of the access in more detail, specifically focusing on single cell colonization and the introduction of new species through contamination of pre-existing microbial communities, (iii) consider experimental bias in methodological approaches, (iv) undertake in situ biodiversity detection to mitigate risk of non-sample return and post-sample contamination, and (v) address the critical question of how important these microbes are in the functioning of Antarctic ecosystems

Buzz off! An evaluation of ultrasonic acoustic vibration for the disruption of marine micro-organisms on sensor-housing materials

Biofouling is a process of ecological succession which begins with the attachment and colonization of micro-organisms to a submerged surface. For marine sensors and their housings, biofouling can be one of the principle limitations to long-term deployment and reliability. Conventional antibiofouling strategies using biocides can be hazardous to the environment, and therefore alternative chemical-free methods are preferred. In this study, custom-made testing assemblies were used to evaluate ultrasonic vibration as an antibiofouling process for marine sensor-housing materials over a 28-day time course. Microbial biofouling was measured based on (i) surface coverage, using fluorescence microscopy and (ii) bacterial 16S rDNA gene copies, using Quantitative polymerase chain reaction (PCR). Ultrasonic vibrations (20 KHz, 200 ms pulses at 2-s intervals; total power 16·08 W) significantly reduced the surface coverage on two plastics, poly(methyl methacrylate) and polyvinyl chloride (PVC) for up to 28 days. Bacterial gene copy number was similarly reduced, but the results were only statistically significant for PVC, which displayed the greatest overall resistance to biofouling, regardless of whether ultrasonic vibration was applied. Copper sheet, which has intrinsic biocidal properties was resistant to biofouling during the early stages of the experiment, but inhibited measurements made by PCR and generated inconsistent results later on

A multi-parametric assessment of decontamination protocols for the subglacial Lake Ellsworth probe

Direct measurement and sampling of pristine environments, such as subglacial lakes, without introducing contaminating microorganisms and biomolecules from the surface, represents a significant engineering and microbiological challenge. In this study, we compare methods for decontamination of titanium grade 5 surfaces, the material extensively used to construct a custom-made probe for reaching, measuring and sampling subglacial Lake Ellsworth in West Antarctica. Coupons of titanium were artificially contaminated with Pseudomonas fluorescens bacteria and then exposed to a number of decontamination procedures. The most effective sterilants were (i) hydrogen peroxide vapour, and (ii) Biocleanse™, a commercially available, detergent-based biocidal solution. After each decontamination procedure the bacteria were incapable of proliferation, and showed no evidence of metabolic activity based on the generation of adenosine triphosphate (ATP). The use of ultraviolet irradiation or ethyl alcohol solution was comparatively ineffective for sterilisation. Hydrogen peroxide vapour and ultraviolet irradiation, which directly damage nucleic acids, were the most effective methods for removing detectable DNA, which was measured using 16S rRNA gene copy number and fluorescence-based total DNA quantification. Our results have not only been used to tailor the Ellsworth probe decontamination process, but also hold value for subsequent engineering projects, where high standards of decontamination are required

Microplastics increase the marine production of particulate forms of organic matter

Microplastics are a major environmental challenge, being ubiquitous and persistent as to represent a new component in all marine environments. As any biogenic particle, microplastics provide surfaces for microbial growth and biofilm production, which largely consists of carbohydrates and proteins. Biofilms influence microbial activity and modify particle buoyancy, and therefore control the fate of microplastics at sea. In a simulated 'plastic ocean', three mesocosms containing oligotrophic seawater were amended with polystyrene microbeads and compared to three control mesocosms. The evolution of organic matter, microbial communities and nutrient concentrations was monitored over 12 days. The results indicated that microplastics increased the production of organic carbon and its aggregation into gel particulates. The observed increase of gel-like organics has implications on the marine biological pump as well as the transport of microplastics in the ocean

Nutrient Limitation in Surface Waters of the Oligotrophic Eastern Mediterranean Sea: an Enrichment Microcosm Experiment

International audienceThe growth rates of planktonic microbes in the pelagic zone of the Eastern Mediterranean Sea are nutrient limited, but the type of limitation is still uncertain. During this study, we investigated the occurrence of N and P limitation among different groups of the prokaryotic and eukaryotic (pico-, nano-, and micro-) plankton using a microcosm experiment during stratified water column conditions in the Cretan Sea (Eastern Mediterranean). Microcosms were enriched with N and P (either solely or simultaneously), and the PO4 turnover time, prokaryotic heterotrophic activity, primary production, and the abundance of the different microbial components were measured. Flow cytometric and molecular fingerprint analyses showed that different heterotrophic prokaryotic groups were limited by different nutrients; total heterotrophic prokaryotic growth was limited by P, but only when both N and P were added, changes in community structure and cell size were detected. Phytoplankton were N and P co-limited, with autotrophic pico-eukaryotes being the exception as they increased even when only P was added after a 2-day time lag. The populations of Synechococcus and Prochlorococcus were highly competitive with each other; Prochlorococcus abundance increased during the first 2 days of P addition but kept increasing only when both N and P were added, whereas Synechococcus exhibited higher pigment content and increased in abundance 3 days after simultaneous N and P additions. Dinoflagellates also showed opportunistic behavior at simultaneous N and P additions, in contrast to diatoms and coccolithophores, which diminished in all incubations. High DNA content viruses, selective grazing, and the exhaustion of N sources probably controlled the populations of diatoms and coccolithophores

Mussel farming in Maliakos Gulf and quality indicators of the marine environment: Good benthic below poor pelagic ecological status

Biological and geochemical variables in the water column and sediments were monitored along a transect of a mussel farm located in a transitional environment in Maliakos Gulf, a semi-enclosed gulf in eastern Mediterranean. Analyses of water, sediment and macrofauna samples were used to calculate ecological status indicators in the context of the European Water Framework Directive. The water column ecological status was "Poor" or "Bad" showing little change with distance from the farm, but the ecological status of the benthic communities was found to be "Good," although there were quantitative changes in macrofaunal indices with distance from the farm. © 2015 Elsevier Ltd

Is in-situ burning an acceptable mitigation option after a major oil spill? Impact on marine plankton

Major oil spills can impose a significant environmental hazard on the marine ecosystem, and a promising mitigation measure is in-situ oil burning (ISB). However, our knowledge of the impact of the burned residues and soot deposition on the marine ecosystem is still limited. We investigated the effects of burned oil residue and soot deposition on the marine plankton communities of the oligotrophic Eastern Mediterranean Sea with a mesocosm experiment. Three triplicated treatments were tested: (1) Iranian crude oil was added and burned (Burned treatment); (2) soot was collected and deposited with artificial rain (Soot); and (3) a non-contaminated Control. Results revealed that Low Nucleic Acid heterotrophic bacteria, Synechococcus spp., and pigmented pico-nano Eukaryotes (pnEuk) were negatively affected in the Burned and Soot treatments. Viruses, heterotrophic pnEuk and ciliates (in Soot) were crucial for controlling the High Nucleic Acid bacteria. Ciliates and most dinoflagellates showed a negative response to the burned residues but were less affected or were even favored when exposed to soot. Our results show that ISB affected the structure and dynamics of the plankton food web through burned residues and soot depositions. However, since the effects appeared at least three days after the ignition, ISB could be combined with subsequent burned residue collection to minimize its impact on the pelagic ecosystem