Optimising AD

Prof JAMES CHONGRoyal Society Industry FellowDepartment of Biology

james.chong@york.ac.uk@insanity_onehttps://goo.gl/c17LxT York 14 August 2019

Wastewater treatmentraw

water

primary sludge

secondary activated sludge

digestate

biogas

AD

final effluent

1% of allUK electricity!

Anaerobic digestionORGANIC POLYMERS

proteins, polysaccharides, lipids

OLIGOMERS & MONOMERSamino acids, sugars, fatty acids

INTERMEDIATESpropionate, butyrate, alcohols

hydrolysis (acidogenesis)

acetogenesis

fermentation

4H2 + CO2 CH4 + 2H2O

4HCOO- + 4H+ CH4 + 3CO2 + 2H2O

CH3OH + H2 CH4 + H2O

methanogenesisCH3COO- + H2O CH4 + HCO3

-

4RCH3 + 2H2O 4RH + 3CH4 + CO2

LON

DON= 107= 1mL AD

measure

understand

modelpredict

manipulate

who?what?why?

Increase solids destructionIncrease gas output

Understand YW’s process

Understand AD microbiology

(YW’s) AD challenges

4 years’ funding from Royal Society to cover 50% salaryAssociated indirect costs covered by University of York

Additional project costs covered by Yorkshire Water

The Industry Fellowship scheme aims to enhance knowledge transfer in science and technology between those in industry and those in academia. It provides opportunities for an academic scientist to work on a collaborative project with industry, or someone employed in industry to work on a collaborative project with a university department or not-for-profit research organisation. It is anticipated that fellows will establish personal and corporate links between the two sectors in the UK as a foundation for their long-term future development.

Royal Society Industry Fellowship

AD Tour de Yorkshire

Bridlington

Naburn

Esholt

Hull

Goole

Sutton

(Mitchell Laithes)

Caldervale

Lundwood

Sandall

Wombwell

Aldwarke

Blackburn Meadows

Woodhouse Mill

Staveley

Old Whittington

Observed speciesA

LLW

LW

_F

WW SA

OW

OW

_F

ST

WM

BM

SB

R_

D1

BR

_D

2H

UG

O_

D1

GO

_D

2E

S_

D1

ES

_D

2E

S_

D3

ES

_D

4C

AS

UN

AB

0

500

1000

1500

2000

2500

3000

ch

ao

1

AL

LW

LW_F

WW

SA

OW

OW_F

ST

WM

BMS

BR_D1

BR_D2

HU

GO_D1

GO_D2

ES_D1

ES_D2

ES_D3

ES_D4

CA

SU

NAB

AL

LW

LW

_F

WW SA

OW

OW

_F

ST

WM

BM

SB

R_

D1

BR

_D

2H

UG

O_

D1

GO

_D

2E

S_

D1

ES

_D

2E

S_

D3

ES

_D

4C

AS

UN

AB

0

500

1000

1500

2000

ob

serv

ed

sp

ecie

s

AL

LW

LW_F

WW

SA

OW

OW_F

ST

WM

BMS

BR_D1

BR_D2

HU

GO_D1

GO_D2

ES_D1

ES_D2

ES_D3

ES_D4

CA

SU

NAB

AL

LW

LW

_F

WW SA

OW

OW

_F

ST

WM

BM

SB

R_

D1

BR

_D

2H

UG

O_

D1

GO

_D

2E

S_

D1

ES

_D

2E

S_

D3

ES

_D

4C

AS

UN

AB

5

6

7

8

9

Sh

an

no

n in

dex

AL

LW

LW_F

WW

SA

OW

OW_F

ST

WM

BMS

BR_D1

BR_D2

HU

GO_D1

GO_D2

ES_D1

ES_D2

ES_D3

ES_D4

CA

SU

NAB

THP

ES_D2

ES_D4

ES_D1

ES_D3

GO_D1

GO_D2

ST

LW_F

OW_F

LW

SU

SA

WM

OW

BMS

AL

WW

CA

NAB

HU

BR_D1

BR_D20

20

40

60

80

10

0

Pro

teobacte

ria

Bacte

roid

ete

s

Pla

ncto

mycete

s

Firm

icute

s

Verru

com

icro

bia

WW

E1

Spiro

chaete

s

Actin

obacte

ria

Chlo

rofle

xi

Synerg

iste

tes

Therm

oto

gae

Arc

haea

NK

B19

OP

9

Lentis

phaera

e

OP

8

Acid

obacte

ria

oth

er (<

0.5

%)

1.0

0.0

0 20 40 60 80 100 0.0 1.0

THP

feed

mes

op

hili

c

alkalinity

CO

D

-0.4 -0.2 0.0 0.2 0.4-0.2

-0.1

0.0

0.1

0.2

0.3

PC1 [27.6%]

PC

2 [

22.8

%]

AL

LW

LW_F

WW

SA

OW

OW_F

ST

WM

BMS

BR_D1

BR_D2

HU

GO_D1

GO_D2

ES_D1

ES_D2

ES_D3

ES_D4

CA

SU

NAB

THP

feedTHP

feed

Correlating chemistry and biology

Additional equipment

x10

= System-60

42 m2

System-60 space

Additional space

Yumechris Amekan(4 year Indonesian Gvt PhD)

Kimberley Barnes(4 year RS Industry PhD)

Atalie Scriverner(RS Industry Summer Studentship)

Matthew Dowson(RS Industry Summer Studentship)

Abdulrahman Khan(4 year Saudi Arabia Gvt PhD)

Sam Hart(RS Industry Summer Studentship)

Additional staff

Annabel Cansdale(MRes student)

Diagnosing problems

normal

foaming

Metabolomics

Sample microbes

Extract DNA

Fragment DNA

218 million pieces!

The world’s worst jigsaw

Build contigs

Metabolism reconstruction

Alessi, Barnes & Chong, in prep

from: Apple Maps, (1/9/17)

1

2

34

from: Apple Maps, (22/5/19)

4 x 1,858m3

Sample pipeline

May Jun JulAug Sep Oct Nov Dec Jan Feb Mar Aprsample

pool subsetof samples

nanopore sequence(1 x pre-release

PromethION flow cell)

DNA extraction(bead beat)

barcodeindividualsamples

short read sequence(Illumina)

Anna Alessi Sally James

30tds d-1 = 4,500m3 CH4 d-1

Nanopore sequencing

512 nanopore channels10-20 Gb / 48 hours

Assembly statssequence

(Gb)# contigs total length

(bp)mean length

(bp)

max length

(bp)

N50

Illumina 203 3,090,644 2,494,872,734 807 539,079 940

Nanopore 60 78,439 1,689,879,228 21,544 1,406,516 38,925

Nanopore + Illumina

203+60 78,439 1,717,415,419 21,895 1,420,398 39,579

1 day 5 hours, 7024 cores105 weeks computeFree (£2.5 million)

14 days, 96 cores192 weeks computeFree* (£6000+£2000)

https://www.cloudsolutions.co.uk/case-study/university-of-york-advance-environmental-research-using-google-cloud-with-cts-sprint/https://edu.google.com/intl/en_uk/why-google/case-studies/university-of-york-gcp/

1 2 3 4 f

CLUSTardCansdale & Chong, in prep

FCB group“bacterium belonging to the WWE1 candidate division […] this bacterium might derive most of its carbon and energy

from the fermentation of amino acids, […] “CandidatusCloacamonas acidaminovorans” is probably a syntrophic

bacterium that is present in many anaerobic digesters”

Pelletier et al (2008) J Bacteriol 190:2572-9DOI: 10.1128/JB.01248-07

Annabel Cansdale

1 2 3 4 5 6 7 8 9 10 11

Methane cm3/ g COD

predicted

1

10

100

1 2 3 4 5 6 7 8 9 10 11

Cluster_k99_599777

Cluster_k99_14128

Cluster_k99_513758

Cluster_k99_227437

Machine learning

Tao et al (2019) Applied Energy 247:670-81

In situ biomethanisation

Microbial changes

Tao et al (2019) Applied Energy 247:670-81

University of SouthamptonUniversity of Sheffield

United UtilitiesLutra

LAMP

Loop-mediated isothermal amplification65°C, 30’

Methanoculleus

Methanothermobacter

Methanococcus

Methanobrevibacter

1 +ve control2-5 mesophilic AD6,7 thermophilic AD8,9 salt marsh10 garden soil11,12 stoolC -ve control (not d)

Alessi & Chong in prep

Future goals

consistency

improvement

enhancement

process

pilot

lab

same performance at all sitesat all times

better than current bestgas output

biomethanisation?higher value products?

James RobsonOliver Noble

Yumechris AmekanKim Barnes

Annabel CansdaleAbdulrahman Khan

Nathan Innard*Anna Alessi

*Henry Nicholls*Farah Shahi

Sam HartCharlotte Wringe

Luna Yuan

Mark BentleySteve Howarth

Peter AshtonSally James

Lesley GilbertJohn Davey

Katherine Newling

Kelly Redeker

ThanksJon Brigg

Tom TaylorPhil Blaen

Rich KershawJason ThorntonGraham Weston

Site Ops at:NaburnEsholt

Blackburn Meadows