Sotirios Vasileiadis

Soil bacterial diversity screening using single 16S rRNA gene V regions coupled with multi-million read generating sequencing technologies.

The novel multi-million read generating sequencing technologies are very promising for resolving the immense soil 16S rRNA gene bacterial diversity. Yet they have a limited maximum sequence length screening ability, restricting studies in screening DNA stretches of single 16S rRNA gene hypervariable (V) regions. The aim of the present study was to assess the effects of properties of four consecutive V regions (V3-6) on commonly applied analytical methodologies in bacterial ecology studies. Using an in silico approach, the performance of each V region was compared with the complete 16S rRNA gene stretch. We assessed related properties of the soil derived bacterial sequence collection of the Ribosomal Database Project (RDP) database and concomitantly performed simulations based on published datasets. Results indicate that overall the most prominent V region for soil bacterial diversity studies was V3, even though it was outperformed in some of the tests. Despite its high performance during most tests, V4 was less conserved along flanking sites, thus reducing its ability for bacterial diversity coverage. V5 performed well in the non-redundant RDP database based analysis. However V5 did not resemble the full-length 16S rRNA gene sequence results as well as V3 and V4 did when the natural sequence frequency and occurrence approximation was considered in the virtual experiment. Although, the highly conserved flanking sequence regions of V6 provide the ability to amplify partial 16S rRNA gene sequences from very diverse owners, it was demonstrated that V6 was the least informative compared to the rest examined V regions. Our results indicate that environment specific database exploration and theoretical assessment of the experimental approach are strongly suggested in 16S rRNA gene based bacterial diversity studies.

Recycled paper-paperboard for food contact materials: Contaminants suspected and migration into foods and food simulant

Contaminant residues in food packaging is a new challenge of our time, as it may pose a threat for consumers. Higher levels of contaminants were observed in food packaging made by recycled materials, even if little information is available for some groups of contaminants. The present study proposes a procedure for analyzing three different groups of organic contaminants in recycled paper and paperboard. Seventeen commercial samples were analyzed for the presence of bisphenol A (BPA), bis (2-ethylhexyl) phthalate (DEHP), nonylphenol monoethoxylate (NMP) and nonylphenol di-ethoxilate (NDP). Not all the samples contained all the contaminants; BPA was the only substance present in all the samples. The concentrations detected were quite high and, in most of the cases, in agreement with results reported in previous studies. Substance migration tests from spiked/non-spiked samples for two dry foods and Tenax® food simulant were undertaken. BPA migration quotients were always lower than 1%, whereas the migration quotients of DEHP were higher than 2.0%. The highest nonylphenols migration quotients were 6.5% for NMP and 8.2% for NDP. Tenax® simulates well the contaminants migration from paperboard to dry food, in some cases being even more severe than the food.

Soil microbial diversity patterns of a lowland spring environment

The Po river plain lowland springs represent unique paradigms of managed environments. Their current locations used to be swamps that were drained 6-7 centuries ago, and they have been in constant use ever since. Our aims were to identify the effects of land use on the microbial communities of these soils, look for associated diversity drivers, and assess the applicability of ecology theories with respect to identified patterns. We screened the microbial diversity across a land use transect via high-throughput sequencing of partial 16S rrRNA gene amplicons. Land use had a major effect on soil properties and microbial community structures. Total organic carbon and pH were major diversity drivers for Bacteria, and pH was important for Archaea. We identified the potential contribution of soil amendments to the indigenous microbial communities, and also gained insights into potential roles of taxa in the organic carbon turnover. Verrucomicrobia coincided with the higher values of the recalcitrant organic carbon. Actinobacteria and Acidobacteria correlated with the more labile organic carbon. Finally, the higher diversity found in the soils less enzymatically active and relatively poorer in nutrients, may be explained to an extent by niche-based theories such as the resource heterogeneity hypothesis and Connells intermediate disturbance hypothesis.

Changes in soil bacterial communities and diversity in response to long-term silver exposure

Silver-induced selective pressure is becoming increasingly important due to the growing use of silver (Ag) as an antimicrobial agent in biomedical and commercial products. With demonstrated links between environmental resistomes and clinical pathogens, it is important to identify microbial profiles related to silver tolerance/resistance. We investigated the effects of ionic Ag stress on soil bacterial communities and identified resistant/persistent bacterial populations. Silver treatments of 50-400 mg Ag kg(-1) soil were established in five soils. Chemical lability measurements using diffusive gradients in thin-film devices confirmed that significant (albeit decreasing) labile Ag concentrations were present throughout the 9-month incubation period. Synchrotron X-ray absorption near edge structure spectroscopy demonstrated that this decreasing lability was due to changes in the Ag speciation to less soluble forms such as Ag(0) and Ag2S. Real-time PCR and Illumina MiSeq screening of 16S rRNA bacterial genes showed β-diversity changes, increasing α-diversity in response to Ag pressure, and immediate and significant reductions in 16S rRNA gene counts with varying degrees of recovery. These effects were more strongly influenced by exposure time than by Ag dose at these rates. Ag-selected dominant OTUs principally resided in known persister taxa (mainly Gram positive), including metal-tolerant bacteria and slow-growing Mycobacteria.

Adaptation of Soil Microorganisms to Trace Element Contamination: A Review of Mechanisms, Methodologies, and Consequences for Risk Assessment and Remediation

The authors provide an updated and integrated view of the adaptation of soil microorganisms to elevated concentrations of trace elements. Starting with a summary of the occurrence of trace elements in soils and their effects on soil microorganisms, the scientific evidence underlying adaptation of microorganisms to trace elements from species to community level is discussed. Insights are given regarding the main physiological processes involved in the resistance of bacteria to toxic elements including the potential importance of horizontal gene transfer in the adaptation process. The review continues with a discussion of how new molecular and biotechnological techniques can enrich this field of study. Scientific evidence is utilized in constructing an illustration of microbial community responses with reference to ecological indicators during various adaptation stages, while the related effects on community biological functionality and resilience are discussed. The authors conclude with an evaluation of the importance of considering adaptation in risk assessment and possible remediation of trace element–contaminated sites.

Manganese and iron as structuring parameters of microbial communities in Arctic marine sediments from the Baffin Bay

The Arctic Baffin Bay between Canada and Greenland is sea ice-covered during the majority of the year, restricting primary production to the summer months. Sediments receive low amounts of mostly terrestrial- and less marine-derived organic matter. To study microbial communities constrained by physicochemical conditions changing with distance from land and ocean depth, we applied high-throughput 16S rRNA gene sequencing and compared sequence diversity with biogeochemical parameters in 40 different sediment samples. Samples originated from seven cores down to 470 cm below seafloor along a shelf-to-basin transect. Bacterial diversity decreased faster with depth in basin than in shelf sediments, suggesting higher organic matter content sustained diversity into greater depths. All samples were dominated by Betaproteobacteria (mostly order Burkholderiales), which were especially abundant in basin sediments with low organic carbon and high Mn and Fe pore water concentrations. Strong statistical correlations between concentrations of reduced Mn and/or Fe and the relative abundances of Betaproteobacteria suggest that this group is involved in metal reduction in Baffin Bay sediments. Dehalococcoidia (phylum Chloroflexi) were abundant in all samples, especially in shelf sediments with high organic content. This study indicates that Mn and/or Fe play important roles structuring microbial communities in Arctic sediments poor in organic matter.

Pesticides water decontamination in oxygen-limited conditions

This study was undertaken to develop a laboratory bioreactor, with a functioning principle similar with that of biobed systems but working in oxygen-limited conditions, suitable for decontaminating wastewater mixtures with pesticides. The system is composed by two cylindrical plastic containers. The first one, where the pesticides solution is collected, is open, whereas the second one, where the biomass is disposed, is closed. The pesticides solution was pumped at the biomass surface and subsequently recollected and disposed in the first container. Four pesticides with different physical-chemical characteristics were tested. The results obtained showed a relatively good capacity of the developed prototype to decontaminate waste water containing the mixture of pesticides. The time of the experiment, the number of cycles that the solution made in the system and the environmental temperature have a significantly influence for the decontamination of acetochlor and chlorpyrifos whereas for the decontamination of terbuthylazine and metalaxyl no significant influence was observed. Even if the present prototype could represent a valid solution to manage the water pesticides residues in a farm and to increase the confidence of bystanders and residents, the practical difficulties when replacing the biomass could represent a limit of the system.