Microbial Biodiversity and Molecular Approach

Biodiversity is given by the variety of species on Earth resulting from billions of years of evolution. Molecular-phylogenetic studies have revealed that the main diversity of life is microbial and it is distributed among three domains: Achaea, Bacteria, and Eukarya. The functioning of whole biosphere depends absolutely on the activities of the microbial world. Due to their versatility microbes are the major natural providers of ecological services as well play major role in semi-artificial systems such as sewage treatment plants, landfills, and in toxic waste bioremediation. As for other organisms many pressures and drivers are causing decrease of microbial biodiversity. Several publications document the effect of chemical pollutants e.g. Polycyclic Aromatic Hydrocarbons (PAHs), of atmospheric pollution, of temperature change and of fertilization on microbial community structure. These studies are now possible because sequencing technologies are in ongoing revolution allowing massive de novo sequencing producing millions of bases in a single day. Metagenomics, metatranscriptomics, metaproteomics and single-cell sequencing are approaches providing a view not only of the community structure (species phylogeny, richness, and distribution) but also of the functional (metabolic) potential of a community because virtually about all genes are captured and sequenced. Unfortunately, although microrganisms are very important for the functioning of whole biosphere public knowledge, awareness and political actions did not yet deal with microbes when biodiversity and its decrease are highlighted.JRC.DDG.H.5-Rural, water and ecosystem resource

Effect of environmental factors and an emerging parasitic disease on gut microbiome of wild salmonid fish

The gastrointestinal tract (GIT) of fish supports a dynamic microbial ecosystemthat is intimately linked to host nutrient acquisition, epithelial development,immune system priming, and disease prevention, and we are far from understandingthe complex interactions among parasites, symbiotic gut bacteria, and host fitness.Here, we analyzed the effects of environmental factors and parasitic burdens on themicrobial composition and diversity within the GIT of the brown trout (Salmo trutta).We focused on the emerging dangerous salmonid myxozoan parasite Tetracapsuloidesbryosalmonae, which causes proliferative kidney disease in salmonid fish, todemonstrate the potential role of GIT micobiomes in the modulation of host-parasiterelationships. The microbial diversity in the GIT displayed clear clustering accordingto the river of origin, while considerable variation was also found among fish fromthe same river. Environmental variables such as oxygen concentration, water temperature,and river morphometry strongly associated with both the river microbialcommunity and the GIT microbiome, supporting the role of the environment in microbialassemblage and the relative insignificance of the host genotype and gender.Contrary to expectations, the parasite load exhibited a significant positive relationshipwith the richness of the GIT microbiome. Many operational taxonomic units(OTUs; n 202) are more abundant in T. bryosalmonae-infected fish, suggesting thatbrown trout with large parasite burdens are prone to lose their GIT microbiome homeostasis.The OTUs with the strongest increase in infected trout are mostly nonpathogenicaquatic, anaerobic sediment/sludge, or ruminant bacteria. Our results underscorethe significance of the interactions among parasitic disease, abiotic factors,and the GIT microbiome in disease etiology.IMPORTANCE Cohabiting microorganisms play diverse and important roles in thebiology of multicellular hosts, but their diversity and interactions with abiotic and bioticfactors remain largely unsurveyed. Nevertheless, it is becoming increasinglyclear that many properties of host phenotypes reflect contributions from the associatedmicrobiome. We focus on a question of how parasites, the host genetic background,and abiotic factors influence the microbiome in salmonid hosts by using ahost-parasite model consisting of wild brown trout (Salmo trutta) and the myxozoanTetracapsuloides bryosalmonae, which causes widely distributed proliferative kidneydisease. We show that parasite infection increases the frequency of bacteria fromthe surrounding river water community, reflecting impaired homeostasis in the fishgut. Our results also demonstrate the importance of abiotic environmental factorsand host size in the assemblage of the gut microbiome of fish and the relative insignificanceof the host genotype and gender.</p

Phylogenetic and Functional Metagenomic Profiling for Assessing Microbial Biodiversity in Environmental Monitoring

Environmental management decisions have to base on holistic understanding of biodiversity and functional capability in ecosystems. Environmental metagenomics is an emerging and powerful approach allowing rapidly and reliably determine and compare microbial biodiversity and functional profiles. Advances in next generation sequencing technologies and bioinformatic tools allow set up analysis pipelines which are useful for well designed and targeted monitoring exercises already today. In the paper we demonstrate how direct sequencing of the total community DNA and analysis of the data are applicable to assess anthropogenic impact on the coastal marine ecosystems.JRC.H.1-Water Resource

Impact of Salinity on the Gastrointestinal Bacterial Community of Theodoxus fluviatilis

Differences in salinity are boundaries that act as barriers for the dispersal of most aquatic organisms. This creates distinctive biota in freshwater and brackish water (mesohaline) environments. To test how saline boundaries influence the diversity and composition of host-associated microbiota, we analyzed the microbiome within the digestive tract of Theodoxus fluviatilis, an organism able to cross the freshwater and mesohaline boundary. Alpha-diversity measures of the microbiome in freshwater and brackish water were not significantly different. However, the composition of the bacterial community within freshwater T. fluviatilis differed significantly compared with mesohaline T. fluviatilis and typical bacteria could be determined for the freshwater and the mesohaline digestive tract microbiome. An artificial increase in salinity surrounding these freshwater snails resulted in a strong change in the bacterial community and typical marine bacteria became more pronounced in the digestive tract microbiome of freshwater T. fluviatilis. However, the composition of the digestive tract microbiome in freshwater snails did not converge to that found within mesohaline snails. Within mesohaline snails, no cardinal change was found after either an increase or decrease in salinity. In all samples, Pseudomonas, Pirellula, Flavobacterium, Limnohabitans, and Acinetobacter were among the most abundant bacteria. These bacterial genera were largely unaffected by changes in environmental conditions. As permanent residents in T. fluviatilis, they may support the digestion of the algal food in the digestive tract. Our results show that freshwater and mesohaline water host-associated microbiomes respond differently to changes in salinity. Therefore, the salinization of coastal freshwater environments due to a rise in sea level can influence the gut microbiome and its functions with currently unknown consequences for, e.g., nutritional physiology of the host

Towards understanding the abundance of non-pollen palynomorphs : A comparison of fossil algae, algal pigments and sedaDNA from temperate lake sediments

Given the increased interest in non-pollen palynomorphs (microscopic objects other than pollen identified from pollen slides) in palaeoecological studies, it is necessary to seek a deeper understanding of the reliability of these results. We combined quantitative information of algal pigments and sedimentary ancient DNA (sedaDNA) of phylotaxonomical resolution to validate the richness and abundance of fossil algae in the sediment of a small temperate lake. For the first time, fossil and sedaDNA algae data were combined in a composite data-set and used to reconstruct algae turnover rates over the last 14,500 years. This comparison serves as both an example of how fossil algae can be used to answer various research questions and as a method of revealing to what extent we can rely on palaeoecological interpretations based solely on the fossil algae record from pollen slides. (C) 2017 Elsevier B.V. All rights reserved.Peer reviewe

Using Microcystin Gene Copies to Determine Potentially-Toxic Blooms, Example from a Shallow Eutrophic Lake Peipsi

Global warming, paired with eutrophication processes, is shifting phytoplankton communities towards the dominance of bloom-forming and potentially toxic cyanobacteria. The ecosystems of shallow lakes are especially vulnerable to these changes. Traditional monitoring via microscopy is not able to quantify the dynamics of toxin-producing cyanobacteria on a proper spatio-temporal scale. Molecular tools are highly sensitive and can be useful as an early warning tool for lake managers. We quantified the potential microcystin (MC) producers in Lake Peipsi using microscopy and quantitative polymerase chain reaction (qPCR) and analysed the relationship between the abundance of the mcyE genes, MC concentration, MC variants and toxin quota per mcyE gene. We also linked environmental factors to the cyanobacteria community composition. In Lake Peipsi, we found rather moderate MC concentrations, but microcystins and microcystin-producing cyanobacteria were widespread across the lake. Nitrate (NO3−) was a main driver behind the cyanobacterial community at the beginning of the growing season, while in late summer it was primarily associated with the soluble reactive phosphorus (SRP) concentration. A positive relationship was found between the MC quota per mcyE gene and water temperature. The most abundant variant—MC-RR—was associated with MC quota per mcyE gene, while other MC variants did not show any significant impact

Using Microcystin Gene Copies to Determine Potentially-Toxic Blooms, Example from a Shallow Eutrophic Lake Peipsi

Global warming, paired with eutrophication processes, is shifting phytoplankton communities towards the dominance of bloom-forming and potentially toxic cyanobacteria. The ecosystems of shallow lakes are especially vulnerable to these changes. Traditional monitoring via microscopy is not able to quantify the dynamics of toxin-producing cyanobacteria on a proper spatio-temporal scale. Molecular tools are highly sensitive and can be useful as an early warning tool for lake managers. We quantified the potential microcystin (MC) producers in Lake Peipsi using microscopy and quantitative polymerase chain reaction (qPCR) and analysed the relationship between the abundance of the mcyE genes, MC concentration, MC variants and toxin quota per mcyE gene. We also linked environmental factors to the cyanobacteria community composition. In Lake Peipsi, we found rather moderate MC concentrations, but microcystins and microcystin-producing cyanobacteria were widespread across the lake. Nitrate (NO3−) was a main driver behind the cyanobacterial community at the beginning of the growing season, while in late summer it was primarily associated with the soluble reactive phosphorus (SRP) concentration. A positive relationship was found between the MC quota per mcyE gene and water temperature. The most abundant variant—MC-RR—was associated with MC quota per mcyE gene, while other MC variants did not show any significant impact

Antibiotic Resistance in Campylobacter spp. Isolated from Broiler Chicken Meat and Human Patients in Estonia

Poultry meat is considered the most important source of Campylobacter spp. Because of ris- ing antimicrobial resistance in Campylobacter spp., this study investigated the antimicrobial resistance of Campylobacter isolates from fresh broiler chicken meat originating from the Baltic countries sold in Estonian retail settings. Additionally, human clinical isolates obtained from patients with Campylobacter enteritis in Estonia were analysed. The aim of this study was to investigate the susceptibility of Campylobacter spp. to nalidixic acid, ciprofloxacin, tetracycline, streptomycin, erythromycin and gen- tamicin. The broth microdilution method with the EUCAMP2 panel was used for MIC determination and antimicrobial mechanisms were analysed using WGS data. A total of 46 Campylobacter strains were analysed, of which 26 (42.6%) originated from Lithuanian, 16 (26.2%) from Latvian, and 4 (6.6%) from Estonian fresh broiler chicken meat. In addition, 15 (24.6%) Campylobacter strains of patients with campylobacteriosis were tested. The antimicrobial resistance patterns of Campylobacter spp. isolated from fresh broiler chicken meat samples of Estonian, Latvian and Lithuanian origin collected in Estonian retail, and from patients with Campylobacter enteric infections, were determined. A total of 46 (75%) of the isolates tested were C. jejuni and 15 (25%) were C. coli. Campylobacter resistance was highest to nalidixic acid (90.2% of strains) and ciprofloxacin (90.2%), followed by tetracycline (57.4%), streptomycin (42.6%) and erythromycin (6.6%). All strains were sensitive to gentamicin. Additionally, antimicrobial resistance genes and point mutations were detected in 27 C. jejuni and 8 C. coli isolates previously assigned as resistant with the phenotypic method. A high antibiotic resistance of Campylobacter spp. in Lithuanian- and Latvian-origin broiler chicken meat and Estonian clinical isolates was found. Similar antibiotic resistance patterns were found for broiler chicken meat and human Campylobacter isolates.This work was supported by the Estonian Research Council grant PRG1441. WORLDCOM project of the One Health European Joint Programme (OHEJP) consortium and received funding from the European Union’s Horizon 2020 Research and Innovation programme [grant number 773830].This work was supported by the Estonian Research Council grant PRG1441. WORLDCOM project of the One Health European Joint Programme (OHEJP) consortium and received funding from the European Union’s Horizon 2020 Research and Innovation programme [grant number 773830]