Anaerobic Digester Microbial Community, Metadata, and Outcomes. Alison Ling, Barr Engineering in collaboration with Microbe Detectives

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1 Anaerobic Microbial Community, Metadata, and Outcomes Alison Ling, Barr Engineering in collaboration with Microbe Detectives

2 Can we give operators more tools to troubleshoot digesters, especially with regard to methane production?

3 Microbiome and Reactor Performance Design Microbiome Performance Outcomes Operation

4 Outline Intro to Microbiology Study Objectives Study Approach and Some Findings Future Steps and Use of Results

5 Microbiome Feedstocks (WAS, primary sludge, high-strength wastes) Hydrolyzers A diverse microbial community is needed to support methanogenesis Acidogens Methanogens CH 4, CO 2, and Water

6 Feedstocks (WAS, primary sludge, high-strength wastes) Microbe Meet and Greet: Hydrolyzers Complex Organics Hydrolyzers

7 Microbe Meet and Greet: Acidogens and Acetogens Complex Organics Hydrogen Acidogens Organic acids

8 Microbe Meet and Greet: Methanogens Hydrogen Organic acids Methanogens CH 4, CO 2, and Water

9 Microbiome Design SRT Reactor type Mesophilic/thermophilic Microbiome Operation Operating ph and temp Feedstocks COD and nutrient loading Chemical additions VFA:Alkalinity Mixing

10 Microbiome and Reactor Performance Design SRT Reactor type Mesophilic/thermophilic Operation Operating ph and temp Feedstocks COD and nutrient loading Chemical additions VFA:Alkalinity Mixing Microbiome What is important here? Performance Outcomes Biogas production COD removal VSS destruction Other operational issues

11 Objectives Help operators improve operations Optimize methane production Minimize downtime Inform full-scale design

12 Study Overview 21 digesters, 60 samples Sample for microbial community 16S rrna gene sequencing by Illumina MiSeq, QIIME Collect operations, design, and performance data

13 Methods Detail Measurable: Who they are related to Taxonomical (ancestry) classification *Results do not definitively indicate metabolic group

14 Methods Detail Measurable: Who they are related to Taxonomical (ancestry) classification YOU ARE HERE *Results do not definitively indicate metabolic group

15 Methods Detail Measurable: Who they are related to Taxonomical (ancestry) classification Hydrolyzers, Acidogens *Results do not definitively indicate metabolic group

16 Methods Detail Measurable: Who they are related to Taxonomical (ancestry) classification Hydrolyzers, Acidogens Methanogens *Results do not definitively indicate metabolic group

17 Methods Detail Measurable: Who they are related to Taxonomical (ancestry) classification Filaments, H 2 S producers *Results do not definitively indicate metabolic group

18 Methods Detail Interest: What they do Hydrolyzers Acidogens and Acetogens Methanogens

19 Microbiome Hydrolyzers Bacteria Acidogens and Acetogens Archaea Methanogens

20 Microbiome 1 February 1 May 1 June 2 February 2 May 2 June

21 Microbiome 1 February 1 May 1 June 2 February 2 May 2 June

22 Findings Design SRT Reactor type Mesophilic/thermophilic Operation Operating ph and temp Feedstocks COD and nutrient loading Chemical additions VFA:Alkalinity Mixing Microbiome What is important here? Performance Outcomes Biogas production COD removal VSS destruction Other operational issues

23 Findings Archaea and Methane

24 Findings Design SRT Reactor type Mesophilic/thermophilic Operation Operating ph and temp Feedstocks COD and nutrient loading Chemical additions VFA:Alkalinity Mixing Microbiome What is important here? Performance Outcomes Biogas production COD removal VSS destruction Other operational issues

25 Findings Community Groupings

26 Findings Community Groupings

27 Feed Source Municipal more diverse

28 Findings Design SRT Reactor type Mesophilic/thermophilic Operation Operating ph and temp Feedstocks COD and nutrient loading Chemical additions VFA:Alkalinity Mixing Microbiome What is important here? Performance Outcomes Biogas production COD removal VSS destruction Other operational issues

29 Findings Archaea and Type

30 Findings Design SRT Reactor type Mesophilic/thermophilic Operation Operating ph and temp Feedstocks COD and nutrient loading Chemical additions VFA:Alkalinity Mixing Microbiome What is important here? Performance Outcomes Biogas production COD removal VSS destruction Other operational issues

31 Temperature Thermophilic reactors have higher methane production and archaea

32 Findings Design SRT Reactor type Mesophilic/thermophilic Operation Operating ph and temp Feedstocks COD and nutrient loading Chemical additions VFA:Alkalinity Mixing Microbiome What is important here? Performance Outcomes Biogas production COD removal VSS destruction Other operational issues

33 COD Removal Directly correlates to methane production

34 Findings Design SRT Reactor type Mesophilic/thermophilic Operation Operating ph and temp Feedstocks COD and nutrient loading Chemical additions VFA:Alkalinity Mixing Microbiome What is important here? Performance Outcomes Biogas production COD removal VSS destruction Other operational issues

35 % Archaea Diversity Micronutrient Addition Correlates to more archaea.... but less diversity No Nutrients Nutrients Added No Nutrients Nutrients Added

36 Key Performance Indices Methane Potential KPI Based on methanogen abundance Community Stability and Diversity KPI Based on community diversity and similarity to reactor baseline Odor KPI Based on abundance of sulfate-reducing bacteria

37 Future Use of Results Provide additional performance metrics Design SRT Reactor type Mesophilic/thermophilic Operation Operating ph and temp Feedstocks COD and nutrient loading Chemical additions VFA:Alkalinity Mixing Microbiome Bio-methane KPI Bio-stability KPI Bio-odor KPI Performance Outcomes Biogas production COD removal VSS destruction

38 Example Key Performance Indicators (KPIs) From Study

39 Future Use of Results Provide additional performance metrics Design SRT Reactor type Mesophilic/thermophilic Operation Operating ph and temp Feedstocks COD and nutrient loading Chemical additions VFA:Alkalinity Mixing Microbiome Bio-methane KPI Bio-stability KPI Bio-odor KPI Requires accurate and continuous monitoring of operating conditions Performance Outcomes Biogas production COD removal VSS destruction

40 Study Report Including Community groupings and relationships to operations and performance parameters Correlations between operations and microbiome Correlations between microbiome and digester performance Description of KPI and how they are derived

41 Performance Dashboard Bio-Methane KPI Example

42 Performance Dashboard Bio-Diversity KPI Example

43 Performance Dashboard AD Output Example Week or Month Normalized KPI CH4 Production COD Removal VSS Destruction Efficiency KPI Efficiency KPI Efficiency KPI % 78% 71% % 73% 69% % 74% 72%

44 Performance Dashboard Example Wee k or Mont h CH4 Production Normalized KPI Efficiency KPI COD Removal Efficiency KPI VSS Destruction Efficiency KPI % 78% 71% % 73% 69% % 74% 72% Bio-Odor KPI Planned

45 Future Work Additional data from more digesters Futher develop KPIs Work with operators to implement lesons learned