Improvement of passive treatment of mine influenced water using genomics

Transcription

1 Improvement of passive treatment of mine influenced water using genomics Susan Baldwin, Marcus Taupp, Parissa Mirjafari, Maryam Khooshnoodi, Steven Hallam Chemical and Biological Engineering, Department of Microbiology and Immunology, University of BriFsh Columbia

2 ObjecFves: Passive treatment is one way of cleaning up mine influenced water so that it can be re used or discharged safely into the environment. The word passive means that treatment of the water is accomplished with natural geochemical and biological processes. This is done by construcfng a system of subsurface and surface flow ponds where the mine water percolates through organic rich material containing microbes that help to remove metals and other chemicals such as sulphate. Some metals and chemicals are removed when the water flows in a verfcal direcfon beneath the surface where there is no oxygen, and other metals are removed when water flows through surface ponds with plants. Because the biological mechanisms taking place inside these passive treatment systems are so complex, we have a hard Fme understanding when they are working or not. To make passive treatment systems more reliable, we need a way of monitoring which microbes are indicafve of a successful treatment system. We are anempfng to accomplish this by sequencing the genomes of the microbes in operafng passive treatment systems.

3 Trail anaerobic bioreactor: ABR1 60% pulp mill biosolids 35% sand 5% cow manure ',(+*5)&,+/"6$%& '/(*&,+0"1$%&!"#$%&'(&)"(*+(#,$-"(&!"#$%&'(&)"*)+*#,$-"*&./0123& A?B5*?:" '#!" CD5*?:" '!!" &!" %!" $!" #!" CD5*?:" '#!" '!!" &!" %!" $!" #!"!" '$()*+(!," A?B5*?:" '$!" '$!"!#(-*.(!%" #$(/01(!%" '2(/03(!%"!#(456(!%" 4$#+& #'(758(!%" '!(9+:(!%" #;(<=>(!%" '$()*+(!,"!#(-*.(!%" #$(/01(!%" '2(/03(!%"!#(456(!%" #'(758(!%" '!(9+:(!%" #;(<=>(!%"

4 Overall removal:!"#$%&'"(')(#*$+"(&,(&-)')./)*& 3"(')(#*$+"(&*)45'+"(&$'*"66&#7)&87"%)&696#).& )!" /:" (!" ;+" '!" &!" %!" $!" #!"!" *+,-." /01"#" /01"$" 2,3+."#" 2,3+."$" 45673" 28+9" 0#*)$.&)1,+(2&

5 The Mount Polley Mine pilot scale treatment pond.

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7 Samples are taken from the treatment system

8 Methodology: Extract samples from the anaerobic bioreactors Extract edna (environmental genomic DNA) Metagenomics Measure water quality from the sites where samples were extracted Correlate metagenomes with environmental condifons

9 Build libraries:

10 Phylum/Class Acidobacteria Actinobacteria Bacteroidetes Chlorobi Cyanobacteria Deferribacteres Fibrobacteres Firmicutes Nitrospirae Planctomycetes!-Proteobacteria "-Proteobacteria #-Proteobacteria $-Proteobacteria Spirochaetes Verrucomicrobia Lentisphaerae Candidate divisions Order/family/genus/ environmental group Acidobacteriales total Bacteroides unclassified Bacteroidales Cytophaga sp. rel. cluster Sphingobacteria Chlorobaculum total Synergistes Fibrobacter Bacillales Mollicutes Clostridia-Veillonellaceae Clostridia-Ruminococcus et rel. Clostridia-Acetobacterium et rel. unclassified Clostridiales Nitrospira total Rhizobiales Rhodobacter Rhodocista Sphingomonas total Acidithiobacillus Aeromonas Rickettsiella Cellvibrio Xanthomonadales Geobacter Desulfobacterales Desulfovibrionales Pelobacter Treponema total total TG3 other Taxon distribution (%): Silage Compost Molasses + hay Wetland biosolids Bacteria that degrade cellulose materials FermentaFve and acetogenic bacteria Sulphate reducing bacteria

11 The comparison organism (assembled confgs from wetland fosmid library; minimum confg length: 2000 bp) were aligned to the reference genome (Methanocorpusculum labreanum Z). The result list the genes of the reference organism in chromosomal order and display hits on the comparison organisms accordingly. Percent protein sequence identity Bidirectional best hit Unidirection al best hit

12 assembled confg 1382 annotafon: 40,789 bp length (marked red in upper window: 16S rdna); lower window: gene organizafon comparison with four different methanogenic reference genomes

13 Batch tests of organic material 450 Sulphate Concentration (mg/l) Time (day) 40% wood chips+30% Hay +30 Manure+ Inoculum 70% Wood chips +15% Hay +15%Manure+Inoculum

14 ConFnuous column tests < 100 mg/l sulfate Synthetic mine drainage, 500 mg/l sulfate

15 Changes in organic maner characterizafon Soluble carbohydrates Recalcitrant components

16 Biosorption Chemical sorption of metal cations via complex formation with cellular ligands(l) M 2 + M 2+ M 2+ As(CH3) Metal (oxidized soluble) 3 (AsO 3 4 ) MO 2+ 2 (As(OH) 3 )MO 2 Metal (reduced insoluble) 2L 2L 2L PO 4 3 transporter s As(OH) 3 M 2+ AsO 3 4 M 2+ Biomineralization Formation of insoluble metal precipitates H2S + AsO 4 3 As2S3 Orpiment Concomitant reducfon of sulfate and arsenate H 2 S + M 2+ MS 2H + M 2+ +CO 3 2 MCO 3 +2OH M(OH) 2 HPO 2 4 +M 2+ MHPO 4 H2S+Fe 2+ FeS pyrite FeS+As FeAsS arsenopyrite Biotransformation Reduction of high valence metal to lower valence insoluble species.