BIOREMEDIATION

Page 173
 Introduction
• This chapter describes the types of contaminant that can be released
into the environment, their structure, fate, and the consequences of the
pollution.
• Strategies for the biotreatment and bioremediation techniques used for
synthetic compounds, petrochemicals, metals, and gaseous wastes are
discussed.
 Synthetic Compounds
• Common synthetic compounds are;
• Pesticides, herbicides, polychlorobiphenyls (PCBs, used in
hydraulic fluids), triclhoroethane (used for wood treatment),
dioxins and dibenzofurans.
• Synthetic compound degrades very slowly (half-lives measured
in years)
 Petrochemical Compounds
• Commonly comes from crude oil and its derivatives (natural gas,
shale oil and tars)
• Compounds in crude oil;
• Methane, aliphatic n-alkanes, monocyclic aromatics; benzene,
polycyclic hydrocarbons; naphthalene, heterocyclic
hydrocarbons, tars and bitumen.
 Inorganic Wastes
• Metals and other inorganic compounds.
• Highly concentrated metals could be toxic towards living
organism.
• Example;
• Copper is an essential micronutrient for plants, but an excess of
copper element could cause inhibition of photosynthesis and
pigment synthesis, and damage to plasma membrane
(Marschner, 1995).
BIOREMEDIATION
STRATEGIES
Genetically Stimulation of
Indigenous indigenous
manipulated microbial
Bioaugmentation Phytoremediation
microorganism
organism growth

Bioremediation strategies
Indigenous micro-organism
• Hydrocarbon-utilizing bacteria and fungi such as cyanobacteria is found in the
soils to degrade hydrocarbons.
• Soil with high level of hydrocarbons contain more microorganisms; less diversity
• Fate of the organic compound depends is affected by two factors:
• Growth and metabolism of microorganism
• Organic compound itself
• Chemical structure of organic compound
• Availability or solubility
• Photochemistry

• Simpler aliphatic or monocyclic aromatics compound degrade easily degrade;

complex structure such as PAHs are hard to degrade.
• Adding surfactant to the soil improves the availability of hydrocarbons and
improves degradation of microorganisms.
Terms for the degradation of organic
compound
• Detoxification : toxic compounds converted into non toxic compounds without
being mineralized
• Example: hydrolysis, dehalogenation, ether cleavage

• Activation : Non-toxic compounds are converted into toxic compounds.

• Example: dichlorination of TCE to produce DCE (harmful) and Vinyl chloride

• Co-metabolism : Breakdown of compound which is not used as a source of energy.

• Degradation of the secondary compound (substrates) depends on the presence of the
primary compound.

• Gratuitous metabolism : Degradation of compound by an unrelated enzyme.

• Due to enzyme not specific with the substrates
STIMULATION OF INDIGENOUS MICROBIAL
GROWTH
• Rate of biodegradation depends on the microbial growth rate
• Factors affecting the microbial growth rate:
• Presence of biodegradable organic material
• Alternative growth substance is needed if the microbes cannot use pollutants as a
energy source.
• Presence of nitrogen and phosphorus
• Add a supplement such as nitrogen and phosphate containing fertilizer
• Oxygen level
• Aerobic degradation of hydrocarbon is faster than anaerobic
• A soil with open structure encourage oxygen transfer
• Temperature
• Each microorganism has a optimum temperature for growth rate
• Low temperature decreases microbial growth rate
• pH level
• pH of the soil depends on the organic compound contain in the soil.
• Adding nitrogen or phosphorus fertilizers can reduce the pH
• Soil moisture
• Type and number of microorganism present
• Presence of heavy metals
• Degradation of xenobiotics (man-made chemicals) depend on the enzymatic
capability of enzymes to degrade the compound.
• Degradation rate is low and period of adaptation is needed
• High level of heavy metals inhibit microbial growth
Bioaugmentation
• Addition of selected organisms to contaminated sites in order to supplement the
indigenous microbial population.
• Factors that causes failure of bioaugmentation.
• Limiting population of organism
• Concentration of contaminant is not sufficient to support growth
• Contain substance that inhibit growth
• Predation by protozoa
• The added microorganisms used some other substance in the environment
• The added microorganisms not able to penetrate through the soil
Modification in bioaugmentation
• Using activated soils
• Contain indigenous microbial populations recently exposed to contaminants
• Introduce naturally developed populations not cultured outside the soil (mixed culture)
• Use fungi as an agent
• The hyphae can penetrate the soil to consume the pollutant
• It can grow under low water condition
• Plasmid-assisted bioaugmentation
• Plasmids contain information of the biodegradation pathway
• The selected bacterial donor containing the desired plasmid is added into the soil
• The donor disappear, but the plasmid was transferred to other microorganism in the soil
which express the genes.
Genetically manipulated organism
• Creation of superbug by genetic manipulation
• Superbug is a genetically modified bacteria to enhance its ability
• Example of superbug:
• Pseudomonas putida is a multiplasmid-containing microorganism that can degrade
lighter alkanes and aromatics.

• Survival of such strain must be questioned

• Issue of releasing into the environment have to be addressed in terms of safety,
containment and public perception
 Bioaugmen-
tation

Bioremedia-
Biosparging
tion on land
Bioremediation
techniques in situ

Land farming Bioventing

Bioremediation on land
• Depends on the:
• Area contaminated
• Properties of the compound
• Concentration of the contaminants
• Required time to complete the bioremediation

• The contaminant can be treated on site or off site

• Pump-and-treat technique is used for water soluble contaminants
• Commonly used in underground water remediation
• Water is introduce to the contaminated areas and removed at another site to be treated
• Frequently used in metal-contaminant treatment
Example of pump-and-treat technique
Land Farming
• Simplest on-site treatment
• Mixing the soil by ploughing/mechanical tilling to increase the oxygen level in the
soil and distribute the contaminants evenly
• Nutrients are added to increase the biodegradation
• Typically 4 to 6 moths to complete the biodegradation process
• Suitable for shallow contamination of the soil surface.
• Treatment area is lined and dammed to retain any contaminated leachate
Bioventing
• Combine increased oxygen supply with vapor extraction
• A vacuum is applied at some depth and a hole is drilled at the site to allow the
airflow
• This will draw the air down into the soil and sweep out any volatile organic
compound
• Nutrient supplies can be provided y running nutrients into trenches
• Only effective for reasonably volatile compound; soil is permeable
 Biosparging
• Increase the biological activity of soil by
increasing the supply of oxygen; sparging air
into the soil.
• The expense of the treatment has limited its
application but on-site generation of oxygen
has reduce its cost
• Hydrogen peroxide is used to replace pure
oxygen but it can be toxic at low
concentration to microorganism
 Biopile
process

Bioremediation
Bioreactors techniques ex Composting
situ

Novel
technologies
Composting
• Composting material such as straw, bark and wood chips are mixed with the
contaminated soil and pile into heaps
• A rise in temperature to 60°C and above, causes by microbial activity
• Higher temperature encourages growth of thermophilic bacteria
• Expensive to be applied at highly contaminated materials
Biopile process
• Soil is heaped into piles within a
lined area to prevent leaching
• The piles are covered with
polythene and liquid nutrients
applied to the surface
• Aeration is improved by applying
suction to the base of the pile
• Any leachate form is collected by
pipes at the base and recycled
 Bioreactor
• Soil extracted from contaminated site can be treated as a solid waste or liquid
leachate in bioreactors
• Easy to control the parameters (pH, temperature, oxygen supply)
• Solid waste slurries can be treated by solid bed, fluidized bed and stirred tank
• Bioreactors for wastewater treatment can be used for treating liquid leachates
• Difference is liquid leachates are low concentration, low loads on system, slow
degradation rates and slow biomass growth
• Immobilized system is the best option

• Activated sludge system is used to remove xenobiotics.

 Bioreactors
• Gives control of the parameters such as temperature, pH,
mixing, and oxygen supply, which can improve degradation
rates.
• Types of bioreactors used as biotreatment;
• Solid-bed, fluidized-bed, stirred-tank, activated sludge, trickling
filter, rotating biological contactor, up-flow anaerobic-sludge
blanket, biopile and rotating drum.
Novel technologies
• Use of specificity of antibody/antigen reaction to remove specific pollutants from
contaminated waters
• Use nanoparticles that will transform wide variety of environmental contaminants
• Permeable reactive barrier used to treat acid mine drainage
• The porous material in the barrier act as a immobilization matrix for
microorganisms to degrade the contaminants
PHYTOREMEDIATION
• Phytoremediation is the use of plants for the removal of contaminants & metals from
soil & water or to render them harmless.

• Advantages use plants for remediation

Aesthetically pleasing option
Has minimal disruption
Has no disruption to topsoil
Effective with low levels of mixed contamination
Offers the possibility of recovery of metals
Inexpensive
• Disadvantages
 Process can be slower than other bioremediation methods
 Taking a number of growing seasons
 Contaminants may reduce the plant growth considerably
 Accumulate pollutants constitute a hazard to wildlife & food chain

• Consideration need to be taken for phytoremediation

 Grow rapidly with high yield per hectare
 Accumulate metal in harvestable parts
 Have profuse root system
 Have high bioconversion factor
• Bioconversion factor is the concentration of pollutant in the plant compared with that in
environment
• Most plant have bioconversion factor less than 1
• Bioconversion for phytoremediation need to be 20 or above to be able to reduce
contamination by 50% in 10 crops
• (Fig. 5.21) clearly shows that the higher the biomass, the more rapid he rate of
phytoremediation
• The plants that accumulate high concentration of metals are known as
hyperaccumulators
• High accumulators must be able to tolerate high levels of metal in their roots & shoots
• Must also be able to take up the metal from the soil at high rates & transfer the metal
from roots to shoots at high rate
• Hyperaccumulation involves adsorption, transport & translocation to area large
quantities of metal can be stored.
 Process in phytoremediation
• Phytoextraction is the uptake of metals & organic pollutants by the roots of
plants & their storage in roots, leaves and stems.
• Plant roots constitute a very large area containing high-affinity chemical
receptors.
• Herbaceous plants are suitable for phytoremediation as they grow rapidly, have
high biomass & can stabilize soils and spoil tips
• Aquatic plants have also been used to take up metals to remove polluted water.
• Trees are a low-cost sustainable & ecologically viable solution for contaminated
land
• The availability of metals & their accumulation varies considerably with trees &
many of investigation have been carried with metal-containing sewage sludge.
• Phytodegradation the uptake & degradation of organic compounds
• Organic compounds can be degraded by plants or sequestered in the vacuole for
degradation later
• Best-known process is the glutathione-S-conjugate transfer system where this
conjugate with organic pollutants
• Organic compound can undergo a number of changes:
Partial transformation in less toxic compound
Partial degradation
Subsequent sequestration
Complete degradation

• Plant contain aliphatic dehalogenases capable degrading TCE

• Plant contain nitroreductases, dehalogenases and laccases can degrade TNT,
RDX and nitroglycerine to carbon dioxide, ammonium and nitrate
• Oxygenation is a common process in pesticide & herbicide degradation which
make molecule more soluble & more suitable for attack.
• Phytovolatilization the volatilization of pollutants into atmosphere
• Some plants can convert metal ions to more volatile species which can reduce
toxicity & aid disposal through stomata
• TCE can be volatilized by poplar, methyl t-butyl ether (MTBE) by eucalyptus,
microorganism associated with plant roots can convert Hg+2 into volatile Hg
• Phytostabilization the transformation of on species of molecule into less-toxic
species or the reduction of mobility
• Green plants have been used to stabilized soils, prevent dispersion of metal
contaminated soil & reduce metal mobility by rhizosphere absorption &
precipitation
• Certain metals & organic contaminants can be concentrated on or in the root
zone without degradation
• Removal of pollutants from the atmosphere which is by taking up the nitrogen
dioxide in atmosphere by Compositae, Mytaceae, Solanaceae & Salicaceae
• Rhizofiltration is the removal of contaminants from flowing water by plant
roots, which can be performed by the roots or microorganism associated with
the roots (rhizosphere) or two combined
• Wetland plants have large population of microorganism associated with their
roots & it is this microbial population is responsible for the sequestration of
heavy metals & breakdown of the organic compounds
• Reed beds has been used to clean up mine leakage containing heavy metals
• Municipal waste water have also been treated using constructed wetlands
• Treatment of wastewater including:
• Water stabilization pond
• Artificial wetland (Typha latifolia)
• Terrestrial section (Salix atrocinerea)
• Rhizostimulation is the stimulation of plant growth by the rhizosphere by
providing better growth conditions or reduction in toxic compunds
• Beneficial free-living soil bacteria are often referred to as growth-promoting
rhizobacteria
• High level of microorganism associated with plant roots is due to high levels of
nutrients that are exuded from the roots which can be up 20% of photosynthetic
yield
• These bacteria can decrease or prevent the effects of pathogens, provide plant
with compound that stimulate growth & help uptake of nutrients
• Phycoremediation is the ultization of macro & microalgae which can be used for
nutrient removal, bioabsorbant, acid & metal wastewater, carbon dioxide
sequestration, biodegradation of xenobiotics & as part of biosensor
• Two microalgal isolates immobilized in such a manner have been shown to
remove ammonium, nitrite and orthophosphate from wastewater using
corrugated raceway bioreactor
• Microalgae such as Chorella vulgaris have been shown to accumulated metals
principally by absorption to the cells surface
Process Summary
METALS & GASEOUS
BIOREMEDIATION
 Metal
Bioadsorption Bioremediation Extracellular Precipitation
- Sulphates removal by using
- Method to remove metals
sulphate-reducing bacteria under
from wastewater
anaerobic condition

Passive uptake Active uptake HOW?

- Rapid (5-10 mins) - Slower than - Bacteria uses simple carbon source to
passive uptake generate hydrogen sulphide from sulphate

- The hydrogen sulphide react with any

- Affected by: - Affected by: metal forming insoluble metal sulphide
• Physical • Metabolic
condition inhibitors
such as pH & • Uncouplers
ionic • Temperature
strength
 Trickling
Biofilters
biofilters

Gaseous
Bioremediation
- Contaminants:
Activated • VOCs
• Sulphur Dioxide Bioscrubbers
sludge
• Nitruous oxides
• CFCs
• Greenhouse effect

Membrane
biorectors
Biofilter
• Can be open or closed system
• Built directly into the ground or in a reactor vessel
• The biofilm is maintained by a continual supply of
nutrients & high humidity by humidifying the inlet
air or by adding nutrient at the top of the vessel.
• Most degradation are aerobic
Trickling Biofilter
• Gas contaminants are absorbed in a free liquid
phase prior to biodegradation by microbes
• Offer greater performance than biofilters at
higher contaminant loadings
• The recycled water can be supplemented with
nutrients, maintaining the viability of the
microbial population
Bioscrubber
• The degradation of contaminants is performed by
a suspended consortium of microbes in separate
vessel
• Absorption achieved in packed column, sprat
tower
• Water transferred to separate vessel, where
optimum environmental conditions for
biodegradation are.
 Biofilter Trickling biofilter Bioscrubber Membrane Activated
bioreactors sludge
- To treat emission that - Primarily used to - Used to remove odour - Remove organic - Treat gaseous
have low concentration remove gases with - An improvement from compounds, suspended wastes by
of contaminates and acidic components biofilter & trickling biofilter matter or biodegradable mixing waste
high gas volume micro pollutants gas stream & air

Advantages: Advantages: Advantages: Advantages: Advantages:

- Low installation cost - Average investment - Not necessary to humidify - Compact - High efficiency
- Cost depending on the and operation cost emissions prior to treat - High effluent quality of pollutant
amount of pre- - pH checking is possible them - High rate of removal (up to
treatment of the under certain - Smaller footprint degradation 99% for
emissions require conditions - pH control & nutrient feed - Able to convert from hydrogen
- High DREs for certain - Biological can be automated existing sulphide)
compounds such as decomposition of - Can treat emissions conventional active
aldehyde, organic acids, components; no VOC containing particulate sludge purification.
nitrous oxide residual products matter
Disadvantage: Disadvantage: Disadvantage: Disadvantage: Disadvantage:
- Larger land - Components with poor - More expensive to install - Aeration limitations - Limited
requirement for solubility are difficult - Over feeding can cause - Membrane pollution adsorption by
conventional design to treat excessive biomass growth - High cost bubble size and
- No continuous internal - More expensive than - Expensive and complex residence time
liquid flow to adjust bed biofilter feeding & neutralizing - Corrosions of
pH or to add nutrient - Packing can become systems the pipework
blocked by biomass
 References
• Cakmak, I., Kurz, H., & Marschner, H. (1995). Short‐term effects of boron,
germanium and high light intensity on membrane permeability in boron deficient
leaves of sunflower. Physiologia Plantarum, 95(1), 11-18.
• Gieg, L. M., Kolhatkar, R. V., McInerney, M. J., Tanner, R. S., Harris, S. H., Sublette,
K. L., & Suflita, J. M. (1999). Intrinsic bioremediation of petroleum hydrocarbons in
a gas condensate-contaminated aquifer. Environmental Science & Technology,
33(15), 2550-2560.
• Zhang, W. X. (2003). Nanoscale iron particles for environmental remediation: an
overview. Journal of nanoparticle Research, 5(3), 323-332.