Ny viden om bakteriers skæbne i renseanlæg. Renser de virkeligt så godt, som vi tror? Per Halkjær Nielsen

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1 Ny viden om bakteriers skæbne i renseanlæg. Renser de virkeligt så godt, som vi tror? Per Halkjær Nielsen CENTER FOR MICROBIALCOMMUNITIES DEPARTMENT OF BIOTECHNOLOGY, CHEMISTRY AND ENVIRONMENTAL ENGINEERING AALBORG UNIVERSITY, DENMARK Modern activatedsludge wastewater treatmentplants with N and P-removal Aalborg Vest Primary settling Process tanks Clarifier Effluent Wastewater Settling Agriculture Return sludge Energy (Methane) Sludge handling Landfill Incineration Reuse/recovery Anaerobic digestion Dewatering 1

2 Top 10 largest plants in the world: 3-8 miope Århus Deer Island Treatment plant, Boston Environmental Biotechnology: manipulation of microbial communities Nitrification Ammonium oxidizers (AOB) Nitrate oxidizers (NOB) Removal of micropollutant Denitrification Denitrifiers Anammox Biological P-removal Polyphosphate accumulating organisms (PAOs) Competitors (GAOs) Anaerobic bacteria Fermenters Iron- and sulfate reducers Removal of pathogens Green house gases N 2 O CH 4 2

3 A holistic approach to study WWTPs and AD 16S rrna amplicon sequencing MiDAS midasfieldguide.org Metagenomics In situ studies The Microbial Database for Activated Sludge Since WWTPs (2-32 samples) samples Design Operational data Microscopy FISH 16S rrna amplicon sequencing 3

4 Identification of bacteria by 16S rrnaamplicon Sequencing Sampling DNA Extraction Sample prep. Sequencing Bioinformatics Microthrix C10_SB1A Tetrasphaera Dechloromonas B45 Trichococcus Rhodoferax P2CN44 Fodinibacter Propionicimonas Hyphomicrobium Simplicispira Gordonia Sphingopyxis Ferruginibacter more Relative abundance [%] Bacteria in nutrient removing WWTP WWTP 2 WWTP 1 WWTP 3? Many common bacteria Few common bacteria 8 4

5 Bacteria in nutrient removing WWTP Many common bacteria WWTP 2 WWTP 1 WWTP 3 9 Each plant has a stable community over time ( ) Albertsen et al., in prep. 16S amplicons 2x250 bp, V1-3 Illumina platform 5

6 Global wastewater treatment system microbiome Very similar bacterial genera/species worldwide Countries cluster separately Primarily due to differences in relative abundances 14 countries >100 samples Marta Nierychlo >230 abundant / process important bacteria 6

7 02/11/2016 Manually curated SILVA 16S rrna taxonomy - proposes a name for all abundant genus-level taxa Link between identity and the available information on phenotypic properties Abundance information based on 10-year-long MiDAS survey McIlroy et al. (2015) MiDAS: the field guide to the microbes of activated sludge. Database, Vol Marta NierychloSimon J. McIlroy Mads Albertsen Bianca McIlroy C ENTER FOR MICROBIAL C OMMUNITIES AALBORG UNIVERSITY Linking identity with function MiDAS overview 7

8 What happens with the influent bacteria? Process tank: Active (important for the treatment), survive inactive or die off Effluent: Most similar to process tank of influent? Pathogens? Incoming bacteria Primary settling Process tanks Settling Effluent Return sludge Energy (Methane) Anaerobic digestion Wastewater treatment in Denmark: no regulation or standards of effluent quality 8

9 Wastewater treatment in Denmark: no regulation or standards of effluent quality By using cultureindependent methods total count is at least 100 greater Sampling in 14 full-scale treatment plants Jannie M. Kristensen 2 Flow proportional samples 1 Grab sample A sample set representing the average microbial community in each plant Influent (+/- primary settling) Aeratedprocess tank Effluent 9

10 1. Influent microbial community composition Top 1 5 Similar over time for all 14 plants 2. Is the Influent community similar to process tanks? Influent Process tank 10

11 Overview: Influent, process tank and effluent Composition in influent among plants more similar than expected Do specific genera takea shortcut through the plants? Some genera are very abundant in effluents compared to process tanks Influent Supernatant Effluent Activated sludgeflocs Excess Biomass Activated sludgeflocs 11

12 Conclusions 1. The influent community composition is surprisingly similar between plants over time and only a few genera makeup a large amount of the biomass 2. Activated sludge: bacteria from influent are mostly not actively growing, but they still account for 3-8 % of the biomass. 3. Some bacteria are taking a shortcut through some plants with the water flow some of them might bepathogenic 4. Is this a problem? Wedon t know yet! May impact the way wastewater treatment plants should berun in the future 5. These studies can only becarried out by using the novel DNA sequencing technologies The MiDAS team Simon J. McIlroy Marta Nierychlo Mads Albertsen Bianca McIlroy Jannie M. Kristensen Rasmus Kirkegaard Martin H. Andersen Søren Karst Mikkel Stokholm-Bjerregaard Marianne Stevenson 12

Aalborg Universitet. Published in: Database. DOI (link to publication from Publisher): /database/bax016. Publication date: 2017

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