BIOREMEDIATION

Bioremediation

Definitions:

- the use of either naturally occurring or deliberately introduced microorganisms or other

forms of life to consume and break down environmental pollutants, in order to clean up a
polluted site.
- is a process that uses mainly microorganisms, plants, or microbial or plant enzymes to
detoxify contaminants in the soil and other environments.
- is an option that enhances the efficacy of the natural biodegradation process of the ocean.
It uses decomposers and green plants, or their enzymes, to improve the condition of
contaminated environment (Atlas & Hazen, 2011).
- uses micro-organisms to reduce pollution through the biological degradation of pollutants
into non-toxic substances. This can involve either aerobic or anaerobic micro-organisms
that often use this breakdown as an energy source.
- is an ecologically sound and state of the art technique that employs natural biological
processes to completely eliminate toxic contaminants. Any process that uses micro
organisms, fungi, green plants or their enzymes to return the natural environment altered
by contaminants to its original condition.
- Geological characteristics of polluted site(s) including soil type, pollutant depth and type,
site location relative to human habitation and performance characteristics of each
bioremediation technique should be incorporated in deciding the most suitable and
efficient method to effectively treat polluted sites.

Bioremediation Approaches

Bioremediation approaches can be applied either in situ or ex situ depending on the nature of

contaminant and site conditions. 

-In situ treatment

is more attractive and cost effective as it is not or less disruptive and does not involve excavation
and transport of contaminated soils. In situ bioremediation techniques have been successfully
used to treat chlorinated solvents, dyes, heavy metals, and hydrocarbons polluted sites (Folch et
al. 2013; Kim et al. 2014; Frascari et al. 2015; Roy et al. 2015). Notably, the status of electron
acceptor, moisture content, nutrient availability, pH and temperature are amongst the important
environmental conditions that need to be suitable for a successful in situ bioremediation to be
achieved (Philp and Atlas 2005). Unlike ex situ bioremediation techniques, soil porosity strongly
influences the application of in situ bioremediation to any polluted site.

*The commonly used in situ approaches include:

 - Natural attenuation

Natural attenuation processes involve contaminant attenuation to harmless products through

natural processes, such as microbial degradation, volatilization, sorption and immobilization. The
natural attenuation process is contaminant specific and commonly employed for petroleum
hydrocarbon contaminated sites. However, natural attenuation may not be a suitable option for
several other contaminants such as persistent organic pollutants. Although natural attenuation has
proven to be a successful approach to treat petroleum contaminants (benzene, toluene,
ethylbenzene and xylene), it may not work if the site does not have the contaminant degrading
microorganisms or nutrients.

- Biostimulation

The microbial transformation of contaminants in soils depends on the availability of nutrients

(carbon, nitrogen, phosphorus and potassium), favourable environmental conditions (pH,
electrical conductivity, aeration, temperature) and the nature of contaminant itself and its
bioavailability. Some contaminants such as persistent organic pollutants (e.g., PAHs, PCBs,
lindane, dichlorodiphynyltrichloroethane) are extremely insoluble in water and tend to strongly
sorb to organic matter in soils thereby decreasing their availability to microbes. The use of
biosurfactants can enhance the bioavailability of such pollutants. The addition of slow release
fertilizers or organic waste and manures can supply the nutrients and stimulate the indigenous
microbes to transform the contaminants.

The addition of natural organic substrates such as mulch and manure has shown to remove
perchlorate through stimulation of anaerobic degradation by microbes (USEPA, 2005).
Perchlorate reducing bacteria are ubiquitous, have the ability to reduce perchlorate to chloride
under anaerobic conditions using perchlorate as a terminal electron acceptor for growth and
energy in the presence of electron donor (Waller et al., 2004). The bioremediation process using
glycerine‐diammonium phosphate (DAP) successfully treated over 160,000 tonnes of soil from a
1000 acre Bermite site from Los Angeles, California containing 0.59–8.4 mg perchlorate/kg soil
to non‐detectable levels within seven month period, which is considered to be a safe and
economical treatment ($35 per tonne). The former Bermite site was used to manufacture various
explosives and related products including perchlorate during 1934–1987 (Evans et al., 2008).

- Bioventing

Bioventing is an in-situ bioremediation process that promotes aerobic biodegradation of organic

contaminants in the unsaturated (vadose) zone. The method is suitable for sites contaminated
with fuel compounds like gasoline, diesel and jet fuel as well as other biodegradable chemicals.
Typically, these pollutants will be biodegraded in aerobic conditions by
indigenous heterotrophic microorganisms naturally occurring in the soil. Thus, in order to
promote microbial degradation, air or poor oxygen is delivered to anaerobic and permeable
polluted soil zones at a low flow rate so the oxygen supply rate meets the demand by the
microorganisms and minimizes volatilization of contaminants.

- Bioaugmentation

Introducing specific microorganisms to decontaminate the soils when indigenous microbes are
not efficient is considered a more acceptable approach to remediate the contaminated soils.
However, the strains for bioaugmentation should ideally have (i) superior ability to degrade the
target contaminants, (ii) easy to cultivate, (iii) fast growth, (iv) tolerance to the high
concentration of contaminant and (v) ability to survive in a wide range of environmental
conditions/stressors. Bioaugmentation has been proven to be successful for a wide range of
pollutants including pesticides such DDT, lindane, endosulfan, pentachlorophenol (PCP),
polyaromatic hydrocarbons (PAHs) and total petroleum hydrocarbons (Abhilash et al., 2011;
Saez et al., 2014; Wang et al., 2014; Chen et al., 2015; Kuppusamy et al., 2016a,b). However,
predation, competition and toxins in soils can negatively affect the survival of introduced
microbes. In such cases, bioaugmentation using immobilized cells in carrier materials or
preadapted strains to the problem soil conditions may prove to be advantageous regarding
enhancing their survival in soils.

- Ex situ approaches

involve excavation and removal of contaminated soil for treatment either on the site or
transportation to a suitable place before treatment.

One of the major advantages of ex situ bioremediation techniques is that they do not require
extensive preliminary assessment of polluted site prior to remediation; this makes the
preliminary stage short, less laborious and less expensive. Due to excavation processes
associated with ex situ bioremediation, pollutant inhomogeneity as a result of depth, non-uniform
concentration and distribution, can easily be curbed by effectively optimizing some process
parameters (temperature, pH, mixing) of any ex situ technique to enhance bioremediation
process. These techniques allow modifications of biological, chemical and physico-chemical
conditions and parameters necessary for effective and efficient bioremediation. Importantly, the
great influence of soil porosity, which governs transport processes during remediation, can be
reduced when polluted soils are excavated. Ex situ bioremediation techniques are unlikely to be
used in some sites such as under buildings, inner city and working sites (Philp and Atlas 2005).
On the other hand, the excavation features of ex situ bioremediation tend to disrupt soil structure;
as a result, polluted and surrounding sites alike experience more disturbances. Moderate to
extensive engineering required for any ex situ bioremediation techniques implies that more
workforce and capital are required to construct any of the technique. In most cases, these
techniques require large space for operation. Generally, ex situ bioremediation techniques tend to
be faster, easier to control and can be used to treat wide range of pollutants (Prokop et al. 2000).

*The commonly used ex situ bioremediation approaches include

- Land farming

Land farming is amongst the simplest bioremediation techniques owing to its low cost
and less equipment requirement for operation. In most cases, it is regarded as ex situ
bioremediation, while in some cases, it is regarded as in situ bioremediation technique. This
debate is due to the site of treatment. Pollutant depth plays an important role as to whether land
farming can be carried out ex situ or in situ. In land farming, one thing is common, polluted soils
are usually excavated and/or tilled, but the site of treatment apparently determines the type of
bioremediation. When excavated polluted soil is treated on-site, it can be regarded as in situ;
otherwise, it is ex situ as it has more in common with other ex situ bioremediation techniques. It
has been reported that when a pollutant lies <1 m below ground surface, bioremediation might
proceed without excavation, while pollutant lying >1.7 m needs to be transported to ground
surface for bioremediation to be effectively enhanced (Nikolopoulou et al. 2013). Generally,
excavated polluted soils are carefully applied on a fixed layer support above the ground surface
to allow aerobic biodegradation of pollutant by autochthonous microorganisms (Philp and
Atlas 2005; Paudyn et al. 2008; Volpe et al. 2012; Silva-Castro et al. 2015). Tillage, which
brings about aeration, addition of nutrients (nitrogen, phosphorus and potassium) and irrigation
are the major operations, which stimulate activities of autochthonous microorganisms to enhance
bioremediation during land farming. Nevertheless, it was reported that tillage and irrigation
without nutrient addition in a soil with appropriate biological activity increased heterotrophic and
diesel-degrading bacterial counts thus enhancing the rate of bioremediation; dehydrogenase
activity was also observed to be a good indicator of biostimulation treatment and could be used
as a biological parameter in land farming technology (Silva-Castro et al. 2015). Similarly, in a
field trial, Paudyn et al. (2008) reported >80 % contaminant (diesel) removal by aeration using
rototilling approach at remote Canadian Arctic location over a 3-year study period; this further
demonstrates that in land farming technique, aeration plays crucial role in pollutant removal
especially at cold regions. Land farming is usually used for remediation of hydrocarbon-polluted
sites including polyaromatic hydrocarbons (Silva-Castro et al. 2012; Cerqueira et al. 2014); as a
result, biodegradation and volatilization (weathering) are the two remediation mechanisms
involved in pollutant removal. Land farming system complies with government regulations, and
can be used in any climate and location (Besaltatpour et al. 2011). The construction of a suitable
land farming design with an impermeable liner minimizes leaching of pollutant into
neighbouring areas during bioremediation operation (da Silva et al. 2012). Over all, land farming
bioremediation technique is very simple to design and implement, requires low capital input and
can be used to treat large volume of polluted soil with minimal environmental impact and energy
requirement (Maila and Colete 2004).

Although the simplest bioremediation technique, land farming like other ex situ bioremediation
techniques has some limitations, which include: large operating space, reduction in microbial
activities due to unfavourable environmental conditions, additional cost due to excavation, and
reduced efficacy in inorganic pollutant removal (Khan et al. 2004; Maila and Colete 2004).
Moreover, it is not suitable for treating soil polluted with toxic volatiles due to its design and
mechanism of pollutant removal (volatilization), especially in hot (tropical) climate regions.
These limitations and several others make land farming-based bioremediation time consuming
and less efficient compared to other ex situ bioremediation techniques.

- Biopiles

Biopile-mediated bioremediation involves above-ground piling of excavated polluted

soil, followed by nutrient amendment, and sometimes aeration to enhance bioremediation by
basically increasing microbial activities. The components of this technique are: aeration,
irrigation, nutrient and leachate collection systems, and a treatment bed. The use of this
particular ex situ technique is increasingly being considered due to its constructive features
including cost effectiveness, which enables effective biodegradation on the condition that
nutrient, temperature and aeration are adequately controlled (Whelan et al. 2015). The
application of biopile to polluted sites can help limit volatilization of low molecular weight
(LMW) pollutants; it can also be used effectively to remediate polluted extreme environments
such as the very cold regions (Dias et al. 2015; Gomez and Sartaj 2014; Whelan et al. 2015). In
line with this, Gomez and Sartaj (2014) studied the effects of different application rates (3 and
6 ml/m3) of microbial consortia, and mature compost (5 and 10 %) on total petroleum
hydrocarbon (TPH) reduction in field-scale biopiles at low temperature conditions, using
response surface methodology (RSM) based on factorial design of experiment (DoE) tone. At the
end of the study period (94 days), 90.7 % TPH reduction in the bioaugmented and biostimulated
setups were obtained compared to the control setups with 48% average TPH removal. The high
percentage of TPH reduction was attributed to synergistic interaction between bioaugmentation
and biostimulation, thus demonstrating the flexibility of biopiles for bioremediation. Similarly,
Dias et al. (2015) reported 71 % reduction in total hydrocarbon concentration, and a shift in
bacterial structure over 50-day study period following pretreatment of contaminated soil samples
prior to biopile formation, and subsequent biostimulation with fishmeal. The feasibility of
biopiles towards bioremediation of different soil samples including clay and sandy soil has been
reported (Chemlal et al. 2013; Akbari and Ghoshal 2014). The flexibility of biopile allows
remediation time to be shortened as heating system can be incorporated into biopile design to
increase microbial activities and contaminant availability thus increasing the rate of
biodegradation (Aislabie et al. 2006). Furthermore, heated air can be injected into biopile design
to deliver air and heat in tandem, in order to facilitate enhanced bioremediation. In another study,
Sanscartier et al. (2009) reported that humidified biopile had a very low final TPH concentration
compared to heated and passive biopiles as a result of optimal moisture content, reduced
leaching, minimal volatilization of less degradable contaminants. In addition, it was reported that
biopile could be used to treat large volume of polluted soil in a limited space. Biopile setup can
easily be scaled up to a pilot system to achieve similar performance obtained during laboratory
studies (Chemlal et al. 2013). Important to the efficiency of biopile is sieving and aeration of
contaminated soil prior to processing (Delille et al. 2008). Bulking agents such as straw, saw
dust, bark or wood chips and other organic materials have been added to enhance remediation
process in a biopile construct (Rodríguez-Rodríguez et al. 2010).

Although biopile systems conserve space compared to other field ex situ bioremediation
techniques, including land farming, robust engineering, cost of maintenance and operation, lack
of power supply especially at remote sites, which would enable uniform distribution of air in
contaminated piled soil via air pump are some of the limitations of biopiles. More so, excessive
heating of air can lead to drying of soil undergoing bioremediation, which will result in
inhibition of microbial activities, and promote volatilization rather than biodegradation
(Sanscartier et al. 2009).

- Windrows

As one of ex situ bioremediation techniques, windrows rely on periodic turning of piled polluted
soil to enhance bioremediation by increasing degradation activities of indigenous and/or transient
hydrocarbonoclastic bacteria present in polluted soil. The periodic turning of polluted soil,
together with addition of water bring about increase in aeration, uniform distribution of
pollutants, nutrients and microbial degradative activities, thus speeding up the rate of
bioremediation, which can be accomplished through assimilation, biotransformation and
mineralization (Barr 2002). Windrow treatment when compared to biopile treatment, showed
higher rate of hydrocarbon removal; however, the higher efficiency of the windrow towards
hydrocarbon removal was as a result of the soil type, which was reported to be more friable
(Coulon et al. 2010). Nevertheless, due to periodic turning associated with windrow treatment, it
may not be the best option to adopt in remediating soil polluted with toxic volatiles. The use of
windrow treatment has been implicated in CH4 (greenhouse gas) release due to development of
anaerobic zone within piled polluted soil, which usually occurs following reduced aeration
(Hobson et al. 2005).
Categories of bioremediation techniques:

SOIL, AIR, AND WATER

Soil

Industrial soils can be polluted by a variety of sources, such as chemical spillages, or the
accumulation of heavy metals from industrial emissions. Agricultural soils can become
contaminated due to pesticide use or via the heavy metals contained within agricultural
products.

A visible example of where bioremediation has been used to good effect can be found in
London’s Olympic Park. The grounds that held the 2012 Olympics had previously been
heavily polluted, after hundreds of years of industrial activity. Bioremediation cleaned 1.7
million cubic metres of heavily polluted soil to turn this brownfield site into one containing
sports facilities surrounded by 45 hectares of wildlife habitats. Groundwater polluted with
ammonia was cleaned using a new bioremediation technique that saw archaeal microbes
breaking down the ammonia into harmless nitrogen gas. The converted park marked the
London 2012 Olympic and Paralympic Games as the “greenest” and most sustainable games
ever held, only possible with bioremediation techniques.

While some soil cleaning techniques require the introduction of new microbes,
‘biostimulation’ techniques increase natural degradation processes by stimulating the growth
of microbes already present. Natural biodegradation processes can be limited by many
factors, including nutrient availability, temperature, or moisture content in the soil.
Biostimulation techniques overcome these limitations, providing microbes with the resources
they need, which increases their proliferation and leads to an increased rate of degradation.

Cleaning up oil-polluted soil is an example of where stimulating microbial growth can be

used to good effect. Research has shown that poultry droppings can be used as a
biostimulating agent, providing nitrogen and phosphorous to the system, which stimulates
the natural growth rate of oil-degrading bacteria. Systems like these may prove cheaper and
more environmentally friendly than current chemical treatment options.

Air

Air is polluted by a variety of volatile organic compounds created by a range of industrial

processes. While chemical scrubbing has been used to clean gases emitted from chimneys,
the newer technique of ‘biofiltration’ is helping to clean industrial gases. This method
involves passing polluted air over a replaceable culture medium containing micro-organisms
that degrade contaminates into products such as carbon dioxide, water or salts. Biofiltration
is the only biological technique currently available to remediate airborne pollutants.
Water

In the UK, access to clean, potable water and modern sanitation is something we take for
granted. However, there are billions of people on Earth for which this is a luxury. The WHO
estimate that each year 842,000 people die as a result of diarrhoeal diseases, many of which
could be prevented if they had access to clean water and proper sanitation. Around 2.6 billion
people lack any sanitation, with over 200 million tons of human waste untreated every year.

Sewage treatment plants are the largest and most important bioremediation enterprise in the
world. In the UK, 11 billion litres of wastewater are collected and treated everyday. Major
components of raw sewage are suspended solids, organic matter, nitrogen and phosphorus.

Wastewater entering a treatment plant is aerated to provide oxygen to bacteria that degrade
organic material and pollutants. Microbes consume the organic contaminants and bind the
less soluble fractions, which can then be filtered off. Toxic ammonia is reduced to nitrogen
gas and released into the atmosphere.

ADVANTAGES AND DISADVAMTAGES OF BIOREMEDIATION

Advantages of bioremediation

Bioremediation is a natural process and is therefore perceived by the public as an acceptable

waste treatment process for contaminated material such as soil. Microbes able to degrade the
contaminant increase in numbers when the contamiant is present; when the contaminant is
degraded, the biodegradative population declines. The residues for the treatment are usually
harmless products and include carbon dioxide, water and cell biomass.

- Theoretically, bioremediation is useful for the complete destruction of a wide variety of

contaminants. Many compounds that are legally considered to be hazardous can be
transformed to harmless products. This eliminates the chance of future liability associated
with treatment and disposal of contaminated material.

- Instead of transferring contaminants from one environmental medium to another, for

example from land to water or air, the complete destruction of target pollutants is possible.

- Bioremediation can often be carried out on site, often without causing a major disruption of
normal activities. This also eliminates the need to transport quantities of waste off site and
the potential threats to human health and the environment that can arise during transportation.
- Bioremediation can prove less expensive than other technologies that are used for clean-up
of hazardous waste.

Disadvantages of bioremediation

Bioremediation is limited to those compounds that are biodegradable. Not all compounds are
susceptible to rapid and complete degradation.
- There are some concerns that the products of biodegradation may be more persistant or
toxic than the parent compound. - Biological processes are often highly specific. Important
site factors required for success include the presence of metabolically capable microbial
populations, suitable environmental growth conditions, and appropriate levels of nutrients
and contaminants.

- It is difficult to extrapolate from bench and pilot scale studies to full-scale field operations.

- Research is needed to develop and engineer bioremediation technologies that are

appropriate for sites with complex mixtures of contaminants that are not evenly dispersed in
the environment. Contaminants may be present as solids, liquids and gases.

- Bioremediation often takes longer than other treatment options, such as excavation and
removal of soil or incineration.

- Regulatory uncertainty remains regarding acceptable performance criteria for

bioremediation. There is no accepted definition of “clean”, evaluating performance of
bioremediation is difficult, and there are no acceptable endpoints for bioremediation
treatments.

Bioremediation is far less expensive than other technologies that are often used to clean
up hazardous waste. There are a number of cost or efficiency advantages to
bioremediation which can be employed in areas that are inaccessible without
excavation.