microbiology in activated sludge processes · 2008-09-17 · wastewater treatment biological...
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©Sara Hallin
Microbiology in
activated sludge
processes
Sara Hallin
SLU
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Wastewater treatment
Biological treatment was introduced in the 1950’s.
The activated sludge processes dominates and is the
largest biotechnology industry in the world.
500 l sewage per person and day in Sweden!
(1,6 x 109 m3/yr)
Historically….
Sanitation problems 1850-1930
Environmental problems 1930-2000
Recycling 2000-
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Microorganisms in activated sludge processes
•Microorganisms are the activated sludge process!
•Bacteria, fungi, viruses, protozoa, algea and metazoa…
•Microorganisms enter the treatment plant from the sewage system.
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Activated sludge processes
• Aeration basin and settling tank.
• The concept: Microorganisms (biomass)
grow in the aeration tank and degrade
organic matter while consuming
oxygen.
• Floc formation is essential for settling.
• The retention time for microorganisms
exceed that for the sewage.
• Nitrogen removal: Biological
• Phosphorus removal: Chemically or
biologically.
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Life on earth
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Litotrophs Organotrophs Heterotrophs
Autotrophs
All organisms
Chemical energy Light energy
Chemotrophs Phototrophs
Autotrophs Mixotrophs
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Metabolism
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Aerobic respiration
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Oxidation:
KOLFÖRENING KOLDIOXID + ELEKTRONER + VÄTEJONER
Reduktion:
SYRE + ELEKTRONER + VÄTEJONER VATTEN
Fullständig reaktion:
KOLFÖRENING + SYRE KOLDIOXID + VATTEN
BIOKEMISKT BUNDEN ENERGI
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Fermentation:
GLUKOS ETANOL + KOLDIOXID
BIOKEMISKT BUNDEN ENERGI
Fermentation av socker till etanol och koldioxid. En del av kolet i
sockret har oxiderats till koldioxid medan en del har reducerats till
etanol (vanlig sprit).
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Aerob Respiration(organotrof)
H20
Fermentation(organotrof)
ATP
8 NADH 2 GTP
6 CO2
Glukos
2 ATP2 NADH
Glukolys
2 Pyruvat
Citronsyracykeln2 FADH
O2
NADH
FADHATP
ATP
Fermentationsreaktioner
Pyruvat
AcCoA
Laktat
Acetate
Etanol
AcetaldehydNADH
NAD+
NADH
NAD+ATP
Redox
+
Glukos
2 ATP2 NADH
Glukolys
2 Pyruvat
-
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Biochemical events in activated
sludge
CO2+H2O+SO42-
PO43-+NO3
-
Organics
(C, H, O, N, P, S)
+
Oxygen
+
Biomass
Mineralised products
Energy (ATP)
Reducing Power
Precursors
Catabolism
Anabolism
Biomass Waste
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Organiskt material bryts ner av mikroorganismer i luftningsbassängen.
Slammet (biomassa och organiskt material)avskiljs från det renade
vattnet i sedimenteringsbassängen..
Traditionell ASP
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NH4+ NO2
- NO3-
N2O
NO
NO2-
Organically bound nitrogen
org-NH2
N2
Nitrogen fixation
Nitrification
Assimilation
AssimilationMineralization/
Ammonification
ATMOSPHERE
SOIL/WATER
Denitrification
Dissimilatory nitrate reduction to ammonium
Microbiological reactions in the N cycle
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Nitrification in water treatment
Nitrification lowers BOD load in recipient
Nitrification coupled to denitrificationlowers N load in recipient
Nitrogen removal systems:- natural wetlands- constructed wetlands- wastewater treatment plants
Process problems with inhibition: Microbial sensors for detectingnitrification inhibitors
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Nitrification
Nitrosomonas
Nitrification involves several reactions andtwo different bacteria:
1. Dissociation of ammonium
2. Oxidation of ammoniaAmmonia monooxygenase (AMO):
Integral membrane proteinCan oxidize methaneCometabolize halogenatedcompoundsInhibited by acetylene
3. Oxidation of hydroxylamineHydroxylamine oxidoreductase (HAO):
Soluble, periplasmic protein
4. Oxidation of nitrite
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Ammonia oxidation:
NH3 + 1,5O2 NO2- + H+ + H2O
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Periplams
OH-
Nitrite oxidation: NO2- + ½ O2 NO3
-
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Carbon metabolism
Cell constituents
Growth
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Energetic constraints...
1. ATP and NADPH (reducing power) requirements in Calvin cycle
2. NAD(P)H formed by reverse e- flow:
Energetic constraints are severe, particularly for NO2- oxidizers.
Many NO2- also grow chemoorganotrophically on glucose or other
substrates.
Cyt c2e-Cyt c
O2
NAD(P)+e-
e-
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Nitrosomonas europeae
The nitrifying bacteria
Key genera: Nitrosomonas, Nitrobacter
Phylogenetically or Proteobacteria(except Nitrospira)
NH3 oxidizers
NitrosomonasNitrosococcusNitrosospira(NitrosolobusNitrosovibrio)
NO2- oxidizers
NitrobacterNitrospinaNitrococcusNitrospira
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Denitrifikation är en andningsprocess som där nitrat
omvandlas till kvävgas i flera komplicerade steg
inom en och samma bakteriecell:
nitrat nitrit kväveoxid lustgas kvävgas
(NO3- NO2
- NO N2O N2)
Styrning och reglering av avloppsreningsverk
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Denitrification: anaerobic respiration
Organic compound CO2
Biosynthesis
Carbon flow
NO3-, (NO2
-, N2O)
Electron flowATP
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Denitrification
Denitrification is a respiratory process where oxidizednitrogen compounds (NO3
-, NO2) are reduced stepwise togaseous end products (NO, N2O, N2):
Cytoplasma
Periplasma
NO3-
NO2- NO3
-
H+
NADH2 NAD+
Proton motive
force
2e- 2e- 2e-
NO2- NO
NO N2O
N2O N2
e-nar nor
nir nos
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Archaea:• Extreme halophiles, Halobacterium
Proteobacteria (α, β, γ):• Phototrophic bacteria, Rhodobacter• Budding bacteria, Hyphomicrobium, Blastobacter• Helical bacteria, Azospirillum, Campylobacter• Nitrogen fixing bacteria, Rhizobium• Ammonia oxidizing bacteria, Nitrosomonas• Gram negative cocci, Paracoccus, Neisseria• Chemoautotrophs, Thiobacillus, Beggiatoa• Others, Alcaligenes, Pseudomonas, Moraxella, Flavobacterium
Gram positives:• Spore forming, Bacillus• Non-spore forming, Corynebacterium
More than 50 genera and 130 species ofdenitrifying bacteria…
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Redox
+
Glukos
2 ATP2 NADH
Glukolys
2 Pyruvat
2 GTP
6 CO2
Citronsyracykeln
8 NADH
2 FADH
ATPNADH
FADHATP
NO3- NO2
- NO N2O N2
-
O2
NH4+
H20
ATP
DenitrifikationNitrifikation
NO2-
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Nitrifierande bakterier Denitrifierande bakterier
•Nitrifierare finns i mark och vatten
•Bara några få arter
•Nitrifikation är två energigivande processer som utförs av två olika grupper av bakterier
•Nitrifierare växer långsamt
•Denitrifierare finns nästan överallt
•Många bakteriesläkten
•Denitrifikation är en alternativ andningsprocess i frånvaro av syre
•Denitrifierare är växer oftast snabbt
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Efterdenitrifikation för biologisk kväveavskiljning
Fördenitrifikation för biologisk kväveavskiljning
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Effekt av extern kolkällapå kvävereningen
0
20
40
60
80
100
0 10 20 30 40 50 60
Nitro
ge
n r
edu
ction
(%)
Time (days)
R
E
a
0
5
10
15
0 10 20 30 40 50 60
Denitri
ficati
on r
ate
(mg
N2O
-N g
-1 V
SS
h-1
)
Time (days)
R
E
b
1. Kvävereningsgrad (%): 2. Denitrifikastionskapacitet:
Tid (dagar) Tid (dagar)
R = Fördenitrifikation utan extern kolkällaE = Fördenitrifikation med etanoltillsats
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Intermittent dosering av etanol
i en fördenitrifikationsprocess
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NH3 NO2- NO3
-
N2O
NO
NO2-
Organic nitrogenorg-NH2
N2
N-fixation
Nitrification
Mineralization
AIR
SOIL/WATER
Denitrification
Dissimilatory nitratreduction to ammonium (DNRA)
N-cycle
Assimilation
Assimilation
N2H2
Anammox
NH2OH
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Anoxic Ammonia Oxidation: Anammox
Uncharacterized organisms from wastewater treatment or nitrate rich sludge have beeen shown to oxidize ammonia to N2 (1995).
The reaction is nitrate or nitrite dependent.
5NH4+ + 3NO2
- 4N2 + 6H2O + 2H+
The organisms grow autotrophically using CO2.
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Ekeby Constructed Wetland
Co
mm
un
itie
s i
n s
ed
ime
nts
Denitrification in a wetland for N-removal
Total area: 36 ha
Flow: ~45000m3/day
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Wa
ter-
flo
w p
ath
s
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11.113.04103.2 NR CTPDA
Multiple regressions:
De
nit
rifi
ca
tio
n r
ate
s
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µg
N/g
DW
/h
Pla
nts
aff
ec
t d
en
itri
fic
ati
on
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Pla
nt
eff
ec
ts i
n
se
dim
en
t
(Ruiz et al., 2008, submitted)
• Typha and Fragmites select nosZ communities
• Seasonal differences
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Biological phosphorus removal
ANAEROBIC AEROBIC
Energy
Phosphate
Short chainfatty acids
EnergyPhosphateO2
CO2+H2O
Energy consumption for uptake of soluble organics.ATP and PO43- is released.
Energy is conserved as poly-phosphate granules. Uptake ofPO4
3-.Consumption of storedproducts (PHB).
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Biological phosphorus removal
PO43-
O2
CO2+H2O EnergyEnergyPO4
3-Organics
Deni
trif
icat
ion
ANAEROBIC REACTOR AEROBIC REACTOR SEDIMENTATION
Recirculation of NO3-
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PHB synthesis & degradation
Acetoacetate ß-hyroxybutyryl-CoA
ß-hydroxybutyrate
Acetoacetyl-CoA
Poly-ß-hyroxybutyrate
(PHB)
Acetic acid
Acetyl-CoA
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Flocken är fundamentet i aktivt slam.
• Snabb och effektiv sedimentering
• Icke sedimenterbara partiklar adorberas
till flocken.
• Biologisk aktivitet är koncentrerad
till flocken.
Sedimentering
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• Slemproducerande bakterier “klistrar” ihop flocken
• Filamentbildande bakterier är flockens “armering”.
• Bakterier är en förutsättning för flockbildning.
• Flockbildande förmåga är självreglerande i systemet.
Flockar skapas av flera olika sorters
mikroorganismer.
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Sedimenteringsproblem som beror på mikroorganismer
Problem Orsak Konsekvens
1. Dispergerad tillväxt Inga riktiga flockar bildas
då mikroorganismerna
förekommer som enskilda
celler
Utgående vatten är
grumligt, dålig
sedimentering
2. Mikroflockar Små, svaga flockar som
lätt slås sönder. Uppstår
då BOD i inkommande
vatten är låg i förhållande
till SS-halten
Utgående vatten är
grumligt, lågt SVI
3. Flytslam Spontan denitrifikation i
sedimenteringsbassängern
a vid långa uppehållstider
Slamtäcke i
sedimenteringsbassänger
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Sedimenteringsproblem som beror på mikroorganismer
Problem Orsak Konsekvens
4. Viskös slamsvällning Mycket extracellulära
polymerer. Vanligt i verk
för industriellt
avloppsvatten med låg
halt av fosfor eller kväve
Dålig sedimentering, slam
i utgående vatten
5. Filamentös
slamsvällning
Mycket filamentbildande
bakterier som
sammanbinder flockar
eller skapar flockar med
hålrum, t ex Microthrix
Högt SVI, och i svåra fall
slamflykt, men klar
vattenfas
6. Skumning Hydrofoba, skumbildande
bakterier tex Nocardia
och Microthrix
Stabilt skum på ytan av
bassänger, slam i
utgående vatten,
skumning i rötkamrarna,
hälsorisk
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Exempel på slamsvällning
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Filamentous bacteria (“Microthrix parvicella”)in activated sludge flocs
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Filamentbildande bakterier
Slamsvällning Skumbildning
”Microthrix parvicella” Microthrix parvicella”
”Nostocoida limicola” Nocardia eller Nocardia amarae-liknande organismer (NALO)
Typ 021N Typ 021N
Typ 0803 Aktinomyceter
Thiothrix
Det finns ca 30 kända
filamentbildare i ASP
10 st står för 90 % av
incidenterna
Geografiska skillnader
Få har "riktiga" namn
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Vad skall man göra?
Lär känna slammet för att hålla koll på filamenten:
1. Bestäm slamvolymindex (SVI)
SVI>150 indikerar risk!
SÄSVSVI 1000
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Vad skall man göra?
2. Mikroskopera slammet regelbundet och bestäm
a) filamenthalt
b) typ av filament
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Filamentbildande mikroorganismer i aktivslamflockar
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Vad skall man göra?
De vanligaste sätten att förska hålla nere filamenthalten är att:
1. Minska slamåldern
2. Öka eller minska luftningen.
3. Införa selektorer/kontaktzoner.
4. Bekämpa med toxiska ämnen.
Ingen av dessa metoder är generellt tillförlitliga.
Kunskapen om filamentbildande bakteriers svar olika parametrar är
bristfällig.
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Minskad filamenthalt efter ozonbehandling
Total Filament Growth
Aug Sep Oct Nov Dec Jan
Jen
kin
s S
cale
1
2
3
4
5
6
Control
Experimental
Extended Filament Growth
Sep Oct Nov Dec Jan
Jen
kin
s S
cale
0
1
2
3
4
5
6
Control
Experimental
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Bättre slamkvalitet – oförändrad N-rening efter ozonbehandling
SQI
Sep Oct Nov Dec Jan Feb
ml/g
0
100
200
300
400
500
600
Control
Experimental
NH4 After Final Settler
Aug Sep Oct Nov Dec Jan Feb
mg
/l0,0
0,5
1,0
1,5
2,0
2,5
3,0
3,5
4,0
Control
Experimental