Environmental Microorganisms

Environmental
Engineering Lab
레 넉 투안
Contents

• General classification

• Microorganisms in the surface waters and

wastewaters

• Applications of Microorganisms to environmental

treatment
Overview: Classification

Classification by
molecular and cellular
relationships:

Bacteria
Prokaryote
Archaea Phylogenetic tree of life

Eukarya Eukaryote
Cell structure

Internal structure of microbial cells.

(a) Diagram of a prokaryote.
(b) Diagram of a eukaryote.
Nucleic acids

Base-pairing in DNA Double helix structure of DNA

Microorganisms in their natural environment

 They don’t live in pure cultures. They live in communities.

 Their activities can greatly affect (change) the chemical and physical properties of
their habitats, e.g. the removal of nutrients and the excretion of waste products.

 Populations in microbial communities can interact in various ways -harmful and/or

beneficial.
Role of microorganisms in geochemical nutrient cycles

• Microorganisms play a role as:

•PRIMARY PRODUCERS
•BIODEGRADERS AND CONSUMERS
• Critical role in cycles of many elements;
• Carbon and and Oxygen cycle
• Nitrogen cycle
• Sulfur cycle
• Phosphorus cycle
How do they grow: requirements?

• Nutrients
• Carbon, Nitrogen, Phosphorus, Sulfur
• Many chemicals supply these
• Micronutrients/ trace metals/ vitamins
• Electron acceptors - usually O2
• Converts / burns carbon substrate to CO2
Energy and biomass * GROWTH
Wastewater treatment: Wastewater treatment: principles
 O2 consumption GROWTH - CELL DIVISION
INCREASE IN BIOMASS
2.0µm
ORGANIC POLLUTANT
AND NUTRIENTS
(C,P,N,O,Fe,S……)

SINGLE
BACTERIUM
CO2
Controlled release of energy evolved
Slow Burning!
Somes microorganisms found in natural waters, wastewaters, and wastewater
treatment processes

Bacteria Archaea Protozoa

Algae Virus
Fungi
 Pathogens and parasites found in domestic wastewater

The three categories of pathogens encountered in the environment are (Leclerc

et al., 2002):

• Bacterial pathogens: Some of these pathogens (e.g., Salmonella,

Shigella) are enteric bacteria. Others (e.g., Legionella, Mycobacterium
avium, Aeromonas) are indigenous aquatic bacteria.
• Viral pathogens: These are also released into aquatic environments but
are unable to multiply outside their host cells. Their infective dose is
generally lower than for bacterial pathogens.
• Protozoan parasites: These are released into aquatic environments as
cysts or oocysts, which are quite resistant to environmental stress and to
disinfection, and do not multiply outside their hosts.
Eg: Giardia lamblia Balantidium coli…
Indicator microorganisms

The total coliform: group belongs to the family enterobacteriaceae and

includes the aerobic and facultative anaerobic, gram-negative, nonspore-
forming, rod-shaped bacteria that ferment lactose with gas production within 48
hours at 35C. This group includes Escherichia coli, Enterobacter, Klebsiella,
and Citrobacter. These coliforms are discharged in high numbers in human and
animal feces, but not all of them are of fecal origin.

Fecal Coliforms: Fecal coliforms or thermotolerant coliforms include all

coliforms that can ferment lactose at 44.5oC. The fecal coliform group
comprises bacteria such as Escherichia coli or Klebsiella pneumonae. The
presence of fecal coliforms indicates the presence of fecal material from warm-
blooded animals.

Escherichia Coli: Several strains of E. coli, many of which are harmless, are
found in the gastrointestinal tract of humans and warm-blooded animals. There
are several categories of E. coli strains, however, that bear virulence factors
and cause diarrhea.
Rivers: problems for surface water resource

• Water quality can be impaired by natural

elements normally found in soil and rock. These
natural elements can be considered
contaminates that can harm humans:
- Metals: arsenic, mercury and lead
- Turbidity: suspended particles of clay soil
- Microorganisms: parasites, bacteria,
protozoa and toxic blue-green algae.

• Faecal matter from confined animal facilities

and feedlots, as well as from animals walking or
defecting in streams, can contaminate water
with bacteria such as E. coli or microbial
pathogens (Cryptosporidium)

• Factory farming of hogs or cattle, where liquid

waste is deliberately forced into the ground
through boreholes can lead to water
contamination.
Municipal wastewater effluents.

• Municipal wastewater effluents or sewage treatment

plants are designed to treat waste that is discharged
by residents, businesses and industries.
• The quality of water that leaves a treatment plant
depends on a variety of factors, such as the type and
efficiency of the treatment and disinfection processes
used, as well as the quantity and types of wastes
received.
• Since treatment processes differ so greatly, municipal
wastewater effluents can consist of residues of human
waste, solids and debris, nutrients, pathogens, toxics,
endocrine disrupting substances, unmetabolized
pharmaceuticals, ingredients from household and
personal care products, and potentially toxic chemicals
and contaminants.
Eutrophication of Surface Waters.c

Natural process in which surface waters receive inputs of nitrates

and phosphates resulting in overnourishment of aquatic ecosystems
(Algae bloom, anaerobic surface water)
• Phosphorus is a key of
nutrient responsible for
eutrophication

The bright green colour results

from bluegreen algae
(Cyanobacteria), which are
growing on phosphorus added to
the near side of the curtain
Microorganisms and Nitrogen

• Nitrogen fixation
Microorganisms and Nitrogen…

• Nitrogen assimilation

Cells convert NO3 or NH4 to proteins and grow until nitrogen becomes
limiting. For each 100 units of carbon assimilated, cells need approximately 10
units of nitrogen (C/N ratio = 10).

• Nitrogen Mineralization
Nitrogen removal

Nitrosomonas, nitrobacter
 Phosphorus (P)

• Organic phosphorus: foodstuff, toilet wastes, animal

wastes…;
• Orthophosphate (PO4-P): fertilizer, products of
phosphorus mineralization;
• Polyphosphate: detergents.
Phosphorus removal

• Chemical precipitation, which is controlled by pH and cations such as

Ca, Fe and Al…
Problems: saturation, clogging.
• Biological phosphorus removal
• Phosphorus assimilation by wastewater microorganisms;
• Enhanced biological phosphorus removal (EBPR);
• Microorganism-mediated enhanced chemical precipitation.
Enhanced biological phosphorus removal (EBPR)

Several microorganisms, called poly P

bacteria or polyphosphate accumulating
organisms (PAOs), have the ability to
accumulate phosphorus in excess of the
normal cell requirement, which is around 1–3
percent of the cell dry weight.

(eg., Acinetobacter, Pseudomonas,

Aeromonas, Moraxella, Klebsiella,
Enterobacter, Tetrasphaera spp.)
Pi release and uptake by a laboratory anaerobic–aerobic
activated sludge. From Hiraishi et al. (1989).
 Enhanced biological phosphorus removal (EBPR)…
Polyhydroxyalkanotes (PHAs) are biodegradable polymers
synthesized by a number of microorganisms when the essential
growth nutrients is limited.
Volatide Fatty
- Under the anaerobic condition, Acid (acetate…)
PAO uptake Volatile fatty acids
(VFAs) mainly acetate and stored in PHA
side the cells as PHA. The energy for
this process is obtained form Anaerobic
condition Glycogen
degradation of polyphosphate P
+ AT
(polyP) and glysosis of glycogen. TCA cycle Poly P
The degradation of poly P results in
the release orthophosphate in the
liquid. o-PO4
Glycogen
- Under the aerobic zone, in the Poly P
Aerobic Cell
absence of any organic compounds, P
PHA is metabolized producing condition AT growth

energy and carbon sources. The PHA

energy is used by PAOs to uptake
orthophosphate in order to recover
O2 CO2 + H2O
and increase the polyP pool in the
cell and to form glycogen Biochemical model (Mino et al., 1987)
 Enhanced biological phosphorus removal (EBPR)…

• Glycogen accumulating organisms (GAOs), are also part of the EBPR

microbial assemblages. (e.g., Candidatus, Competibacter, phosphatis).
Although GAOs carry out carbon transformations similar to those of PAOs,
they do not take up or release phosphorus (Blackall et al., 2002).

• For the successful operation of the EBPR process one must create
conditions for the predominance of PAOs over GAOs. One such condition
is the operation of the EBPR process at pH higher than 7 (Filipe et al.,
2001) since GAOs predominate at lower pHs. Low temperature is another
factor controlling the predominance of PAOs over GAOs. The
psychrophilic PAOs are generally favored at low temperatures, leading to
an increase in EBPR efficiency (Erdal et al., 2003).
Biological waste-water treatment: The activated
sludge process.
 Sludge Microbiology

• Bacteria: Zooglea, Pseudomonas, Flavobacterium, Alcaligenes, Achromobacter,

Corynebacterium, Comomonas, Brevibacterium, Acinetobacter, Bacillus…
• Fungi: Geotrichum, Penicillium, Cephalosporium, Cladosporium, and Alternaria…
• Protozoa: Aequorea victoria, Aspidisca costata, Cryptosporium…
Products from Microorganisms: Overview of range of
examples.

• Various foods and drinks

• Enzymes for varied uses (GM enzymes); biocatalysts
• Engineered proteins ( antibodies )
• Vaccines and antibiotics (secondary metabolites)
• Primary metabolites and bulk chemicals (amino acids (glutamic acid) and
organic acids (acetic acid)
• Pharmaceuticals and novel chiral chemicals
• Recovery of metals in bioleaching
• Biosensors (use of enzymes to specifically detect chemicals in medical)
Thank you for your attention!