Diversity and Cyclical Seasonal Transitions in the Bacterial Community in a Large and Deep Perialpine Lake

High-throughput sequencing (HTS) was used to analyze the seasonal variations in the bacterioplankton community composition (BCC) in the euphotic layer of a large and deep lake south of the Alps (Lake Garda). The BCC was analyzed throughout two annual cycles by monthly samplings using the amplification and sequencing of the V3–V4 hypervariable region of the 16S rRNA gene by the MiSeq Illumina platform. The dominant and most diverse bacterioplankton phyla were among the more frequently reported in freshwater ecosystems, including the Proteobacteria, Cyanobacteria, Bacteroidetes, Verrucomicrobia, Actinobacteria, and Planctomycetes. As a distinctive feature, the development of the BCC showed a cyclical temporal pattern in the two analyzed years and throughout the euphotic layer. The recurring temporal development was controlled by the strong seasonality in water temperature and thermal stratification, and by cyclical temporal changes in nutrients and, possibly, by the remarkable annual cyclical development of cyanobacteria and eukaryotic phytoplankton hosting bacterioplankton that characterizes Lake Garda. Further downstream analyses of operational taxonomic units associated to cyanobacteria allowed confirming the presence of the most abundant taxa previously identified by microscopy and/or phylogenetic analyses, as well as the presence of other small Synechococcales/Chroococcales and rare Nostocales never identified so far in the deep lakes south of the Alps. The implications of the high diversity and strong seasonality are relevant, opening perspectives for the definition of common and discriminating patterns characterizing the temporal and spatial distribution in the BCC, and for the application of the new sequencing technologies in the monitoring of water quality in large and deep lakes.

[1]  Rob Knight,et al.  Microbial Biogeography of Public Restroom Surfaces , 2011, PloS one.

[2]  A. Klindworth,et al.  Evaluation of general 16S ribosomal RNA gene PCR primers for classical and next-generation sequencing-based diversity studies , 2012, Nucleic acids research.

[3]  B. Satinsky,et al.  Bacterial Biogeography across the Amazon River-Ocean Continuum , 2017, Front. Microbiol..

[4]  Zhi-Luo Deng,et al.  Co-occurrence Analysis of Microbial Taxa in the Atlantic Ocean Reveals High Connectivity in the Free-Living Bacterioplankton , 2016, Front. Microbiol..

[5]  Ubiquity and quantitative significance of bacterioplankton lineages inhabiting the oxygenated hypolimnion of deep freshwater lakes , 2016, bioRxiv.

[6]  N. Salmaso,et al.  Planktic Tychonema (Cyanobacteria) in the large lakes south of the Alps: phylogenetic assessment and toxigenic potential. , 2016, FEMS microbiology ecology.

[7]  N. Salmaso,et al.  Expansion of bloom-forming Dolichospermum lemmermannii (Nostocales, Cyanobacteria) to the deep lakes south of the Alps: Colonization patterns, driving forces and implications for water use , 2015 .

[8]  Danilo Ercolini,et al.  High-Throughput Sequencing and Metagenomics: Moving Forward in the Culture-Independent Analysis of Food Microbial Ecology , 2013, Applied and Environmental Microbiology.

[9]  Susan Holmes,et al.  phyloseq: An R Package for Reproducible Interactive Analysis and Graphics of Microbiome Census Data , 2013, PloS one.

[10]  Yu Hen Hu,et al.  A decade of seasonal dynamics and co-occurrences within freshwater bacterioplankton communities from eutrophic Lake Mendota, WI, USA , 2012, The ISME Journal.

[11]  R. Knight,et al.  UniFrac: a New Phylogenetic Method for Comparing Microbial Communities , 2005, Applied and Environmental Microbiology.

[12]  E. Plummer,et al.  A Comparison of Three Bioinformatics Pipelines for the Analysis ofPreterm Gut Microbiota using 16S rRNA Gene Sequencing Data , 2015 .

[13]  J. Pernthaler,et al.  Ecology and Distribution of Thaumarchaea in the Deep Hypolimnion of Lake Maggiore , 2015, Archaea.

[14]  Ben Nichols,et al.  VSEARCH: a versatile open source tool for metagenomics , 2016, PeerJ.

[15]  Pelin Yilmaz,et al.  The SILVA ribosomal RNA gene database project: improved data processing and web-based tools , 2012, Nucleic Acids Res..

[16]  Vincent J. Denef,et al.  Are Oligotypes Meaningful Ecological and Phylogenetic Units? A Case Study of Microcystis in Freshwater Lakes , 2017, Front. Microbiol..

[17]  P. Legendre,et al.  Ecologically meaningful transformations for ordination of species data , 2001, Oecologia.

[18]  Michael Zeder,et al.  A small population of planktonic Flavobacteria with disproportionally high growth during the spring phytoplankton bloom in a prealpine lake. , 2009, Environmental microbiology.

[19]  Y. Igarashi,et al.  Distinct Network Interactions in Particle-Associated and Free-Living Bacterial Communities during a Microcystis aeruginosa Bloom in a Plateau Lake , 2017, Front. Microbiol..

[20]  W. Liesack,et al.  Opitutus terrae gen. nov., sp. nov., to accommodate novel strains of the division 'Verrucomicrobia' isolated from rice paddy soil. , 2001, International journal of systematic and evolutionary microbiology.

[21]  W. Vyverman,et al.  Bacterial Community Composition in Lake Tanganyika: Vertical and Horizontal Heterogeneity , 2005, Applied and Environmental Microbiology.

[22]  Marti J. Anderson,et al.  A new method for non-parametric multivariate analysis of variance in ecology , 2001 .

[23]  Luis Pedro Coelho,et al.  Structure and function of the global ocean microbiome , 2015, Science.

[24]  I. Jasser,et al.  Picocyanobacteria: The Smallest Cell-Size Cyanobacteria , 2017 .

[25]  Simon Garnier,et al.  Default Color Maps from 'matplotlib' , 2015 .

[26]  Brian C. Thomas,et al.  The human gut and groundwater harbor non-photosynthetic bacteria belonging to a new candidate phylum sibling to Cyanobacteria , 2013, eLife.

[27]  David P. Hamilton,et al.  Derivation of lake mixing and stratification indices from high-resolution lake buoy data , 2011, Environ. Model. Softw..

[28]  M. Höfle,et al.  Isolation of Novel Ultramicrobacteria Classified as Actinobacteria from Five Freshwater Habitats in Europe and Asia , 2003, Applied and Environmental Microbiology.

[29]  O. White,et al.  Environmental Genome Shotgun Sequencing of the Sargasso Sea , 2004, Science.

[30]  Gianluca Corno Effects of nutrient availability and Ochromonas sp. predation on size and composition of a simplified aquatic bacterial community. , 2006, FEMS microbiology ecology.

[31]  S. Toshchakov,et al.  Structure of the archaeal community in the Black Sea photic zone , 2015, Microbiology.

[32]  N. Salmaso,et al.  Diversity and distribution of cyanobacterial toxins in the Italian subalpine lacustrine district , 2012 .

[33]  T. Hansen Bergey's Manual of Systematic Bacteriology , 2005 .

[34]  J. Humbert,et al.  Planctomycetes in Lakes: Poor or Strong Competitors for Phosphorus? , 2013, Applied and Environmental Microbiology.

[35]  A. Eiler,et al.  Composition of freshwater bacterial communities associated with cyanobacterial blooms in four Swedish lakes. , 2004, Environmental microbiology.

[36]  Xi Xiao,et al.  Use of High Throughput Sequencing and Light Microscopy Show Contrasting Results in a Study of Phytoplankton Occurrence in a Freshwater Environment , 2014, PloS one.

[37]  S. Amalfitano,et al.  Grazing-induced Synechococcus microcolony formation: experimental insights from two freshwater phylotypes. , 2016, FEMS microbiology ecology.

[38]  V. Blinov,et al.  Bacterial community composition in the water column of the deepest freshwater Lake Baikal as determined by next-generation sequencing. , 2016, FEMS microbiology ecology.

[39]  Nico Salmaso,et al.  Limnological research in the deep southern subalpine lakes: synthesis, directions and perspectives , 2010 .

[40]  M. Figueras,et al.  Population dynamics and ecology of Arcobacter in sewage , 2014, Front. Microbiol..

[41]  R. Stocker,et al.  Zooming in on the phycosphere: the ecological interface for phytoplankton–bacteria relationships , 2017, Nature Microbiology.

[42]  P. Servais,et al.  Bacterial Community Composition in Three Freshwater Reservoirs of Different Alkalinity and Trophic Status , 2014, PloS one.

[43]  Duccio Cavalieri,et al.  MICCA: a complete and accurate software for taxonomic profiling of metagenomic data , 2015, Scientific Reports.

[44]  A. Eiler,et al.  Diurnal variations in the auto- and heterotrophic activity of cyanobacterial phycospheres (Gloeotrichia echinulata) and the identity of attached bacteria , 2006 .

[45]  H. D. Laughinghouse,et al.  On the use of high‐throughput sequencing for the study of cyanobacterial diversity in Antarctic aquatic mats , 2016, Journal of phycology.

[46]  A. Walsby,et al.  Stratification by cyanobacteria in lakes: a dynamic buoyancy model indicates size limitations met by Planktothrix rubescens filaments. , 2005, The New phytologist.

[47]  Philip Abraham,et al.  An Improved Method for High Quality Metagenomics DNA Extraction from Human and Environmental Samples , 2016, Scientific Reports.

[48]  J. Pernthaler,et al.  Succession of bacterial grazing defense mechanisms against protistan predators in an experimental microbial community , 2005 .

[49]  C. Bernard,et al.  Appendix 2: Cyanobacteria Associated With the Production of Cyanotoxins , 2017 .

[50]  J. Pernthaler,et al.  Bloom of Filamentous Bacteria in a Mesotrophic Lake: Identity and Potential Controlling Mechanism , 2004, Applied and Environmental Microbiology.

[51]  Rudolf Amann,et al.  Comparative 16S rRNA Analysis of Lake Bacterioplankton Reveals Globally Distributed Phylogenetic Clusters Including an Abundant Group of Actinobacteria , 2000, Applied and Environmental Microbiology.

[52]  Paramvir S. Dehal,et al.  FastTree 2 – Approximately Maximum-Likelihood Trees for Large Alignments , 2010, PloS one.

[53]  Jonathan Friedman,et al.  Inferring Correlation Networks from Genomic Survey Data , 2012, PLoS Comput. Biol..

[54]  F. Pomati,et al.  Sedimentary DNA Reveals Cyanobacterial Community Diversity over 200 Years in Two Perialpine Lakes , 2016, Applied and Environmental Microbiology.

[55]  H. Grossart,et al.  Enrichment and cultivation of pelagic bacteria from a humic lake using phenol and humic matter additions. , 2010, FEMS microbiology ecology.

[56]  W. Van Criekinge,et al.  Bacterial Diversity Assessment in Antarctic Terrestrial and Aquatic Microbial Mats: A Comparison between Bidirectional Pyrosequencing and Cultivation , 2014, PloS one.

[57]  N. Salmaso,et al.  Global expansion of toxic and non-toxic cyanobacteria: effect on ecosystem functioning , 2015, Biodiversity and Conservation.

[58]  D. Fontaneto,et al.  Distribution patterns and environmental correlates of Thaumarchaeota abundance in six deep subalpine lakes , 2015, Aquatic Sciences.

[59]  N. Salmaso Interactions between nutrient availability and climatic fluctuations as determinants of the long-term phytoplankton community changes in Lake Garda, Northern Italy , 2011, Hydrobiologia.

[60]  K. Šimek,et al.  Limnohabitans curvus gen. nov., sp. nov., a planktonic bacterium isolated from a freshwater lake. , 2010, International journal of systematic and evolutionary microbiology.

[61]  Donald A. Jackson PROTEST: A PROcrustean Randomization TEST of community environment concordance , 1995 .

[62]  L. McNoe,et al.  Characterization of the cyanobacteria and associated bacterial community from an ephemeral wetland in New Zealand , 2016, Journal of phycology.

[63]  C. Borrego,et al.  Phylogenetic characterization and quantification of ammonia-oxidizing archaea and bacteria from Lake Kivu in a long-term microcosm incubation. , 2013, International microbiology : the official journal of the Spanish Society for Microbiology.

[64]  Z. Quan,et al.  Analysis of the bacterial communities associated with different drinking water treatment processes , 2013, World Journal of Microbiology and Biotechnology.

[65]  N. Salmaso,et al.  Long-term trends and fine year-to-year tuning of phytoplankton in large lakes are ruled by eutrophication and atmospheric modes of variability , 2012, Hydrobiologia.

[66]  J. Stockner,et al.  Freshwater Picocyanobacteria: Single Cells, Microcolonies and Colonial Forms , 2012 .

[67]  N. Salmaso,et al.  Frequent recombination shapes the epidemic population structure of Planktothrix (Cyanoprokaryota) in Italian subalpine lakes , 2013, Journal of phycology.

[68]  D. Sigee Freshwater Microbiology: Biodiversity and Dynamic Interactions of Microorganisms in the Aquatic Environment , 2005 .

[69]  Philip Hugenholtz,et al.  A renaissance for the pioneering 16S rRNA gene. , 2008, Current opinion in microbiology.

[70]  J. Ravel,et al.  Evaluation of Methods for the Extraction and Purification of DNA from the Human Microbiome , 2012, PloS one.

[71]  in chief George M. Garrity Bergey’s Manual® of Systematic Bacteriology , 1989, Springer New York.

[72]  A. Eiler,et al.  Flavobacteria Blooms in Four Eutrophic Lakes: Linking Population Dynamics of Freshwater Bacterioplankton to Resource Availability , 2007, Applied and Environmental Microbiology.

[73]  H. Bürgmann,et al.  Estimating Bacterial Diversity for Ecological Studies: Methods, Metrics, and Assumptions , 2015, PloS one.

[74]  R. Stepanauskas,et al.  Comparative single-cell genomics reveals potential ecological niches for the freshwater acI Actinobacteria lineage , 2014, The ISME Journal.

[75]  T. Jurczak,et al.  First report of cyanobacterial paralytic shellfish toxin biosynthesis genes and paralytic shellfish toxin production in Polish freshwater lakes , 2017 .

[76]  R. Stepanauskas,et al.  Metabolic potential of a single cell belonging to one of the most abundant lineages in freshwater bacterioplankton , 2012, The ISME Journal.

[77]  W. Carmichael,et al.  Isolation and characterization of hepatotoxic microcystin homologs from the filamentous freshwater cyanobacterium Nostoc sp. strain 152 , 1990, Applied and environmental microbiology.

[78]  George M. Garrity,et al.  The Archaea and the deeply branching and phototrophic bacteria , 2001 .

[79]  Cedric E. Ginestet ggplot2: Elegant Graphics for Data Analysis , 2011 .

[80]  H. Oh,et al.  Algae-bacteria interactions: Evolution, ecology and emerging applications. , 2016, Biotechnology advances.

[81]  M. Simona,et al.  A synoptic study of phytoplankton in the deep lakes south of the Alps (lakes Garda, Iseo, Como, Lugano and Maggiore) , 2003 .

[82]  Manuel Porcar,et al.  Unveiling Bacterial Interactions through Multidimensional Scaling and Dynamics Modeling , 2015, Scientific Reports.

[83]  Victor Markowitz,et al.  Complete genome sequence of Pirellula staleyi type strain (ATCC 27377T) , 2009, Standards in genomic sciences.

[84]  D. Debroas,et al.  Temporal dynamics and phylogenetic diversity of free-living and particle-associated Verrucomicrobia communities in relation to environmental variables in a mesotrophic lake. , 2013, FEMS microbiology ecology.

[85]  N. Salmaso,et al.  Morpho-Functional Groups and phytoplankton development in two deep lakes (Lake Garda, Italy and Lake Stechlin, Germany) , 2007, Hydrobiologia.

[86]  T. Fenchel The microbial loop -25 years later , 2008 .

[87]  N. Salmaso,et al.  Biogeography of bloom-forming microcystin producing and non-toxigenic populations of Dolichospermum lemmermannii (Cyanobacteria). , 2017, Harmful algae.

[88]  T. Nõges,et al.  Contrasting seasonal and interannual environmental drivers in bacterial communities within a large shallow lake: evidence from a seven year survey , 2015 .

[89]  Haneul Kim,et al.  Sediminibacterium goheungense sp. nov., isolated from a freshwater reservoir. , 2014, International journal of systematic and evolutionary microbiology.

[90]  D. Dietrich,et al.  Anatoxin-a producing Tychonema (Cyanobacteria) in European waterbodies. , 2015, Water research.

[91]  C. Callieri,et al.  Long-term trends of epilimnetic and hypolimnetic bacteria and organic carbon in a deep holo-oligomictic lake , 2010, Hydrobiologia.

[92]  J. Humbert,et al.  Spatiotemporal Changes in the Structure and Composition of a Less-Abundant Bacterial Phylum (Planctomycetes) in Two Perialpine Lakes , 2011, Applied and Environmental Microbiology.

[93]  Atsushi Kouzuma,et al.  Exploring the potential of algae/bacteria interactions. , 2015, Current opinion in biotechnology.

[94]  T. Weisse Pelagic Microbes ‐ Protozoa and the Microbial Food Web , 2007 .

[95]  Georgios A. Pavlopoulos,et al.  Metagenomics: Tools and Insights for Analyzing Next-Generation Sequencing Data Derived from Biodiversity Studies , 2015, Bioinformatics and biology insights.

[96]  G. Garrity Bergey’s Manual® of Systematic Bacteriology , 2012, Springer New York.

[97]  Anders F. Andersson,et al.  Freshwater bacterioplankton richness in oligotrophic lakes depends on nutrient availability rather than on species–area relationships , 2011, The ISME Journal.

[98]  W. Fischer,et al.  Crown group Oxyphotobacteria postdate the rise of oxygen , 2017, Geobiology.

[99]  N. Salmaso,et al.  Taxonomic Identification of Cyanobacteria by a Polyphasic Approach , 2017 .

[100]  B. Ibelings,et al.  Toxic cyanobacteria and cyanotoxins in European waters – recent progress achieved through the CYANOCOST Action and challenges for further research , 2017 .

[101]  E. Reavie,et al.  Ice cover extent drives phytoplankton and bacterial community structure in a large north-temperate lake: implications for a warming climate. , 2016, Environmental microbiology.

[102]  F. Barbosa,et al.  Seasonality of freshwater bacterioplankton diversity in two tropical shallow lakes from the Brazilian Atlantic Forest , 2017, FEMS microbiology ecology.

[103]  Philip Hugenholtz,et al.  NAST: a multiple sequence alignment server for comparative analysis of 16S rRNA genes , 2006, Nucleic Acids Res..

[104]  Katherine D. McMahon,et al.  A Guide to the Natural History of Freshwater Lake Bacteria , 2011, Microbiology and Molecular Reviews.

[105]  H. Grossart,et al.  Particle-Associated Differ from Free-Living Bacteria in Surface Waters of the Baltic Sea , 2015, Front. Microbiol..

[106]  James R. Cole,et al.  Ribosomal Database Project: data and tools for high throughput rRNA analysis , 2013, Nucleic Acids Res..

[107]  Anders F. Andersson,et al.  Transitions in bacterial communities along the 2000 km salinity gradient of the Baltic Sea , 2011, The ISME Journal.

[108]  F. Hildebrand,et al.  Diversity of toxin and non-toxin containing cyanobacterial mats of meltwater ponds on the Antarctic Peninsula: a pyrosequencing approach , 2014, Antarctic Science.

[109]  Hongjuan Yuan,et al.  Sediminibacterium salmoneum gen. nov., sp. nov., a member of the phylum Bacteroidetes isolated from sediment of a eutrophic reservoir. , 2008, International journal of systematic and evolutionary microbiology.

[110]  E. Delong,et al.  The Microbial Engines That Drive Earth's Biogeochemical Cycles , 2008, Science.

[111]  Sharon L. Grim,et al.  Oligotyping: differentiating between closely related microbial taxa using 16S rRNA gene data , 2013, Methods in ecology and evolution.

[112]  M. Sogin,et al.  Minimum entropy decomposition: Unsupervised oligotyping for sensitive partitioning of high-throughput marker gene sequences , 2014, The ISME Journal.

[113]  J. Chun,et al.  Introducing EzBioCloud: a taxonomically united database of 16S rRNA gene sequences and whole-genome assemblies , 2017, International journal of systematic and evolutionary microbiology.

[114]  R. Parsons,et al.  Minor revision to V4 region SSU rRNA 806R gene primer greatly increases detection of SAR11 bacterioplankton , 2015 .