Basic biological

processes
Introduction
1. Introduction
Basic, yes. But for whom?

For us! In our microbial manipulation!

Microbes may disagree - .

 Suggested topics:
Henze Østgaard
1997 2005 Subject Presented by .
3.1 5.1 Intro Kjetill Ø
3.2 5.2 Hydrolysis Marit
3.3 5.3 Aerobic heterotrophic - trivial ?
3.4 5.4 Nitrification Tu
3.5 5.5 Denitrification Andrea
5.6 Sulfur circus - complex ?
3.6 5.7 Phosphate accumul. Torunn
3.7 5.8 Anaerobic fermentation - after lunch
5.9 Toxicity & inhibition - process?
5.10 Predation Øystein
3. Aerobic heterotrophic growth
Compare with
anaerobic scheme:

Note high yield; Aerobic catabolism

Ysx ≈ 0.50 (wght/wght) Water +

≈ 0.65 (COD/COD)
 Metabolic diversity
Metabolic classification of selected basic biological processes:
Energy
source e-donor C source e-acceptor Comment

Photo-

Chemo- organo- organo* O2 5.3 Aerobic heterotroph

litho- NH4+ CO2 O2 5.4 Nitrification

Chemo- organo- organo* NO3- 5.5 Denitrification

+
litho- NH4 CO2 NO2- Anammox

Chemo- organo- organo** SO42-, S0 5.6 S cirkus SRB

litho- H2S CO2/(org.**) O2/(NO3-) SOB

Chemo- no external organo** no external 5.7 Bio P uptake PHB

internal internal O2/NO3- growth

Chemo- no external organo* no external 5.8 Anaerobic fermentstion

* including 5.2 Hydrolysis. ** lacking ability of hydrolysis.

6. Sulfur circus
Too many redox steps:
H2S → S0 (S8) → H2SO4

Too many compounds:

SO2, SO32-, S2O32-, SO42-

Connected to Fe2+/Fe3+.

Activated sludge S cycle: →

8. Anaerobic fermentation
See case in next
session:
9. Toxicity and inhibition
Everything is toxic!*
Kuenen, on crossing of
Monod curves; - may be
competitive advantage! * It is only a question of dose →

Modelling by ”switch”
1,2

1 Monod
0,8
Ks Monod
my

function →0 0,6
Switch
Ks+Cs 0,4

Cs 0,2 Switch
Monod K +C → 1 0
s s 0 20 40 60 80 100 120
Cs
Hydrolysis
 Marit’s file was not available
NITRIFICATION

Tu Anh Vo
 How does the nitrogen cycle work?
 Step 1- Nitrogen Fixation- Special bacteria convert the
nitrogen gas (N2 ) to ammonia (NH3) which the plants can
use.
 Step 2- Nitrification- Nitrification is the process which
converts the ammonia into nitrite ions which the plants can
take in as nutrients.
 Step 3- Ammonification- After all of the living
organisms have used the nitrogen, decomposer bacteria
convert the nitrogen-rich waste compounds into simpler
ones.
 Step 4- Denitrification- Denitrification is the final step in
which other bacteria convert the simple nitrogen compounds
back into nitrogen gas (N2 ), which is then released back
into the atmosphere to begin the cycle again.
How does the nitrogen cycle work?
Nitrification
Nitrification is a microbial process by
which reduced nitrogen compounds
(primarily ammonia) are sequentially
oxidized to nitrite and nitrate.
The process is performed by
chemolithotrophic bacteria that are
capable of growing on reduced
inorganic nitrogen compounds

It involves the transfer of eight

electrons to the electron
transport chain which fuels the
synthesis of ATP.
Inorganic nitrogen

Ammonia, nitrite, and nitrate are all highly soluble in water.

Ammonia exists in two forms: un-ionized NH3, and ionized
NH4. with different concentration depend on pH, temperature
and salinity.
The sum of the two (NH4+ + NH3) is called total ammonia
or simply ammonia.
Inorganic nitrogen compounds: NH4+ - N (ionized ammonia
nitrogen), NH3–N (un-ionized ammonia nitrogen), NO2–N
(nitrite nitrogen) and NO3–N (nitrate nitrogen).
The oxidation of ammonia (NH3)
to nitrite (NO2-) and nitrate (NO3-)

Two phylogenetically distinct groups of bacteria:

Autotrophic bacteria
 Ammonia oxidizing bacteria obtain their energy by
catabolizing un-ionized ammonia to nitrite - Nitrosomonas,
Nitrosococcus, Nitrosospira, Nitrosolobus, and Nitrosovibrio.
 Nitrite oxidizing bacteria oxidize nitrite to nitrate,
Nitrobacter, Nitrococcus, Nitrospira, and Nitrospina.

Heterotrophic bacteria have been shown to oxidize NH3 (and

some organic N) to NO3- - Arthrobacter, Thiosphaera
 The oxidation of ammonia (NH3)
to nitrite (NO2-) and nitrate (NO3-)

Autotrophic bacteria - Nitrification is a two-step process,

normally carried out sequentially and the first step has a
higher kinetic reaction rate than the second step

Heterotrophic bacteria

Most energy is produce and most of the oxygen is consumed

Factors affecting nitrification

• pH

• Alkalinity

• Temperature

• Oxygen

• Salinity

• Light
 Factors affecting nitrification
The most recent results suggest that the optimum range of pH
for nitrification can range from 7.0 to 9.0 .The optimum pH for
Nitrosomonas ranges from 7.2 to 7.8 and from 7.2 to 8.2 for
Nitrobacter.

Alkalinity is a measure of the buffering capacity of an aquatic

system. Protons released during the oxidation of ammonia effect
the buffer capacity. If the alkalinity is not high enough (at least
1.5 – 2mmol/l at the end of nitrification), a significant decrease
in pH can occur
Temperature plays a significant role in the nitrification
reaction rate in suspended growth systems as it does in all
chemical and biological kinetic reactions
Factors affecting nitrification

For every gram of ammonianitrogen oxidized to nitrate-

nitrogen, 4.57 g of oxygen is required. Nitrosomonas very
little at DO levels above 2.0 mg/L, but Nitrobacter exhibited a
reduced growth rate at DO levels of less than 4 mg/L.
(Dissolved Oxygen - DO)
There is very limited information on the impact of
salinity on nitrification. Salinity is similar to both
temperature and pH, in that nitrifying bacteria can
acclimate to almost any salinity range, given sufficient
time.
Light has been shown to inhibit growth of the bacteria
and can encourage the growth of algae on the biomedia.
Thank you
Andrea Bagi, 2008
 Conversion of nitrate into atmospheric nitrogen

Ared + NO3- Aox + 0.5 N2

 Also known as dissimilatory nitrate reduction

 Anoxic organotrophic
microorganisms

-No oxygen (or very low levels)

-Organic carbon as substrate
and C-source
 Energy yielding process

 Substrate removal rate

 Energy sources (substrate)

 Temperature („the higher the better”)

 Oxygen (electron acceptor)

 pH (7-9)

 Nutrients (NH4-N, P)
 Endogenous denitrification in clarifier
◦ Floating sludge

 Measuring the metabolic activity

◦ Nitrogen Uptake Rate (NUR)
 Enhanced biological
phosphorous removal
-EBPR

Torunn Forberg
 Phosphorous (P)
• Main component of many detergents

• Fertilizers (organic and inorganic)

contain high amounts of P

• P input critical in eutrophication

 Wastewater treatment
• P can be removed chemically..

• More “environmentally friendly” to do it

biologically

• Based on microorganisms who

accumulate P
 Phosphorous accumulating
organisms (PAO)
• PAO bacteria can store P as poly-P
granules

• Under good conditions (feast) they store

organic polymers, PHA or PHB, and
release phosphate

• Famine – stored PHA is used for energy

and phosphate is accumulated
 General set up
Anaerobic/Aerobic

University of Cape Town (UCT)

N2
Metabolism of PAO
“Feast” “Famine”
 Who are these PAO?
• Acinetobacter?

• Rhodocyclus related “Accumulibacter

phosphatis”

• Molecular techniques reveal

phylogenetically diverse communities
EBPR’s big problem, the GAO (?)

• EBPR systems are unstable/unreliable

• Glycogen accumulating bacteria (GAO)

• Not quite sure what happens.. means

chemicals are often still used as backup
removal of P
EBPRs big problem, the GAO
Picture from: “Putative glycogen-
accumulating organisms belonging to the
Alphaproteobacteria identified through
rRNA-based stable isotope probing”
Rikke Louise Meyer , Aaron Marc Saunders and
GAO! Linda Louise Blackall

Other ref:
“The microbiolgy of phosphorous removal in
activated sludge processes – the current state of
play” Robert J. Seviour and Simon McIlroy

“Miljøbioteknologi” Del I Basis Kap. 5

Østgaard (2005)
PAO
38

Predation in wastewater
treatment
• Definition
• Important groups
• Effects of predation on wastewater treatment
39

Predation
• Predation (def.): The consumption of one organism
by another, where the consumed organism is alive
when the consumer first attacks it (Begon et al 1990)
40

Dominating grups:
• Protozoa: • Metazoa:
- Flagellates - Nematoda
- Rhizopods - Rotifera
- Actinopods - ”Water bears” (Tardigrada)
- Ciliates: - Higher microfauna
free living
crawling
attached (stalked)
41

Flagellates
• Flagellates: 1-8 flagellas, small (5-40
µm)
– Some osmotrophs, some phagotrophs
– Examples: Pleuromonas sp. (image),
Bodo sp.
Monosiga sp.
42

Rhizopoda
• Big (50-400 µm), flexible
• Moving by pseudopodia
• Amoebae (upper image)
• Thecoamoebae (lower image)
43

Actinopoda
• 40-100 µm
• Thin, needlelike
pseudopodia
• Uses the pseudopodia to
capture prey
44

Ciliates
• Varying size (5-1000 µm)
• Cilia for capturing food and
locomotion
• Free living: Paramecium sp.,
Paramecium sp.
Spirostum sp.
• Crawling: Aspidisca sp.
• Attached: Vorticella sp.,
Carchesium sp.
• Characteristic protozoa of
activated sludge

Vorticella sp.
Aspidisca sp.
45

Metazoa
• Nematodes (500-3000 µm)
• Rotifera (40-500 µm)
• ”Water bears”, Tardigrada
(Vannbjørn) (100-1500 µm)
• Other metazoans: Insects,
(birds, foxes)
46

Effects of predation in
wastewater treatment
• Increased flocculation of bacteria
• Removal of dispersed bacteria
• Increase of food/microorganism (F/M) ratio by
reducing the number of bacteria
• Direct uptake of substrates
• Sludge reduction
• Indicator organisms
47

Litterature
Begon, M., Harper, J.L., Townsend, C.R., (1990): ”Ecology: individuals,
populations and communities”, Oxford: Blackwell Scientific Publications
The Biology of Activated Sludge: http://www.tvt-bio.com/micro2.html
Østgaard, K., (2005): ”Miljøbioteknologi” Del I: Basis.
Wanner, J., (1997): ”Microbial community analysis: The key to the design
of biological wastewater treatment system” Cambridge, University
Press
Luxmy, B.S., Nakajima, F., Yamamoto, K., (2000): ”Predator grazing effect
on bacterial size distribution and floc size variation in membrane-
separation activated sludge” Water Science and Technology, Vol 42
Nos 3-4, pp 211-217