CONTENTS

1) ABSTRACT 04

2) INTRODUCTION 05

3) BIOREMEDIATION 07
3.1) CLASSES OF BIOREMEDIATION 07
3.2) FACTORS RESPONSIBLE FOR MICROBIAL BIOREMEDIATION 08
3.3) TYPES OF POLLUTANTS 09

4) BIOFILMS 10
4.1) Formation of Biofilms 10
4.2) Extracellular Polymeric Substances (EPS) 12

5) BIOFILMS IN BIOREMEDIATION 14

6) BIOFILMS AND ORGANIC POLLUTANTS 17

6.1) Polycyclic Aromatic Hydrocarbons (PAHs) 17
6.2) Polychlorinated Ethenes (PCEs) 18
6.3) Polychlorinated Biphenyls (PCB) 19
6.4) Nitroaromatic Compounds 19
6.5) Pharmaceutical & Personal Care Products (PPCPs) 20

7) LIMITATIONS OF BIOFILMS 21

8) CONCLUSION 22

9) REFERENCES 23

1
LIST OF FIGURES AND TABLES
FIG 1) Factors Influencing Bioremediation 08
FIG 2) Mycoplasma pneumonae biofilm on glass, 50 degree tilt,
20K magnification, by scanning electron microscopy (electron
micrograph scanned by Steven Distelhorst, Miami University) 10
FIG 3) Steps involved in biofilm formation 11
FIG 4) Bacillus subtilis cells grown on nutritive agar plates on
(a) complex medium (LB) and
(b) biofilm-promoting minimal medium. 12
FIG 5) Polycyclic Aromatic Hydrocarbons: structures of some examples 17
FIG 6) Reductive dechlorination of polychlorinated ethenes by catabolic
organohalogen respirations. Two physiologically distinguishable
groups of bacteria mediate this process. 18
FIG 7) Structure of Polychlorinated Biphenyls 19
FIG 8) Nitroaromatic compounds: structures of some examples. 20
FIG 9) Structure of Triclosan 20

TABLE 1) Major properties of biofilms helpful in bioremediation

and the characteristics associated with it 15

2
ABSTRACT
The process of using organisms to neutralize or remove contaminants or pollutants from a
media is known as bioremediation. It is a cost effective and environment friendly process
that uses microorganisms to decontaminate and degrade a wide variety of contaminants
into less harmful products. As compared to free-floating planktonic cells, microorganisms
that exist inside biofilms are more advantageous for bioremediation as they have greater
tolerance to pollutants, environmental stress and ability to degrade very harsh pollutants
through diverse catabolic pathways.
This paper highlights the beneficial use of microbial biofilms in bioremediation of
environmental pollutants.

3
INTRODUCTION
The introduction of substances in the natural environment which are harmful or toxic to it
is known as pollution. These substances which cause pollution are known as pollutants.
Pollutants can take many forms, they can be chemical in nature or they can be in the form
of energy (like noise, heat or light), they can be foreign substances or naturally occurring
substances.

The first form of pollution was air pollution and it started in the prehistoric times when
man created the first fires. According to an article in the journal of Science in 1983, soot
found on the ceilings of prehistoric caves provides enough evidence of the high levels of
pollution that was associated with insufficient ventilation of open fires.

Pollution became a popular issue in America after the World War 2, due to radioactive
fallout from atomic testing and warfare. Then a non-nuclear event, The Great Smog of
1952 in London killed at least four thousand people. This prompted some of the first
major modern environmental legislation, The Clean Air Act of 1956.

For decades, industrialization has created waste all around the world, resulting in
contaminated deposits in soil and their consequent far-reaching diffusion into the aquatic
environment. Apart from causing environmental distress (such as global warming, ocean
acidification, acid rain, biomagnification of heavy metals etc.) pollution also leads to
adverse effects on human health.

Environmental remediation are processes by which the pollutants or contaminants of

environmental media such as soil, ground water, sediment or surface water are removed.
Traditional methods of remediation include dredging, capping, surfactant enhanced
aquifer remediation (SEAR), etc. These processes mainly include removal of
contaminants or sediments to designated waste sites using various means. These process
reduce the risk of exposure to indigenous organism but they are very expensive processed
which only dilute the problem. Even though these processes do provide some initial
relief, the pollutants still remain in the environment, causing harm in the long run.

Owing to their extremely toxic nature even at trace concentrations, these environmental
pollutants are a big threat to our life and environment. As persistent pollutants have
accumulated in the environment for decades and new types are being created every
second, the incentive to apply an appropriate solution to efficiently decontaminate and
reduce the exposure in the environment is even greater.

4
Bioremediation is an emerging technology used to treat contaminated media like water,
soil and surface material, by altering environmental conditions to stimulate growth of
microorganisms which degrade the target pollutants. This biological process for treating
toxic deposits is much better than chemical and physical methods economically and also
in terms of its efficiency. The potential of biofilm communities for bioremediation
process has recently been realized.

A biofilm is a syntrophic association of microorganisms in which the cells stick to each

other and often also to a surface. These cells may include a single species of bacteria to a
combination of different species of bacteria, protozoa, fungi and algae. These adherent
cells become embedded within a slimy extracellular matrix that is composed of
extracellular polymeric substance (EPS). This EPS provides the microorganisms within
the biofilm extra strength to survive under stress conditions under which their free
floating counter-parts might degenerate.

This paper discusses biofilms in relation to their suitability in application of

bioremediation of persistent organic pollutants (POPs).

5
BIOREMEDIATION
The process of using organisms to neutralize or remove contaminants or pollutants from a
media is known as bioremediation. This form of waste remediation uses no toxic
chemicals, although the organisms used can be harmful under certain, very specific,
conditions.

Bioremediation is beneficial over traditional remediation process for three main reasons,
among many other:

1. Chemical free: One of the drawbacks of using chemicals to treat and breakdown
pollutants is that the chemicals eventually make their way into the water supply.
2. Recycling: bioremediation allows for the waste to be recycled. Once the waste is
treated and the contamination neutralized or removed, the waste itself can be
recycled.
3. Economically beneficial: bioremediation, for the most part, involved promoting
the growth of microorganisms already present in the waste to be treated. So it cuts
the cost of the various chemicals used in the traditional remediation process. There
is also no translocation of the waste required as bioremediation can be done in-
situ. There are no involvements of specialized machines which also cuts the cost
of the entire process substantially.

CLASSES OF BIORMEDIATION
Bioremediation is divided into two classes based on the general application of the
organisms being used. These two classes are:

1. In-situ Bioremediation: When the contaminated waste is treated right at its points
of origin, the bioremediation process is known as in-situ. There is no translocation
of the medium involved, and the entire process takes place at the site of the
pollution. In situ treatment prevents the spreading of contamination during the
displacement and translocation of the contaminated material.
2. Ex-situ Bioremediation: When the contaminated waste is removed and
transported into a separate, off-site treatment area, the remediation process is
known as ex-situ. This process helps to contain and control the bioremediation
products, as well as, it makes the area that was contaminated available for further
use.

6
FACTORS RESPONSIBLE FOR MICROBIAL BIOREMEDIATION
The use of microbes for bioremediation is known as microbial bioremediation. A major
advantage of this bioremediation method is that it allows for the contamination to be
treated, neutralized or removed and then produces a waste product that is more easily
disposed off. And in some cases, there is no need for the disposal at all since the product
produced is not harmful or waste.

The factors responsible for this kind of remediation process are:

1. Presence of suitable microorganisms: The presence of a population of the

suitable microorganisms that can biodegrade all of the contaminants is very
important for the bioremediation process. If such microorganisms are absent, they
can be introduced into the polluted environment as well, but only after doing a
thorough research of the effects of its introduction into the environment.
Otherwise, the bioremediation process is done ex-site.
2. Oxygen: there should be enough oxygen present to support aerobic biodegradation
(about 2% oxygen in the gas phase in soil or 0.4 mg/L in water.)
3. Water: the moisture content in soil is very important for the microbes to grow and
biodegrade the pollutants. Soil moisture should be between 50 to 70 % of the
water holding capacity of soil.
4. Nutrients: All microbes need nutrients to grow. The polluted environment or
media should not lack the essential nutrients for the growth of the microorganisms
like nitrogen, phosphorous, sulfur and other nutrients to support good microbial
growth.
5. Temperature: The temperature needs to be appropriate for the specific
microorganisms that need to grow for the bioremediation process.

7
 6. pH: the pH also needs to be appropriate for the growth of specific
microorganisms.

TYPES OF POLLUTANTS
For the effective use of bioremediation on contaminated waste, it is very important to
study the nature of the pollutants in the waste. As a material or substance breaks down it
nature, it produces Figure 1: Factors influencing bioremediation
many different
substances. As it moves through
its stages of decomposition,
varying levels of toxic substances may be produced.

Humans are capable of producing an enormous amount of pollutants is a well-known

fact. Remedying these pollutants with unnatural or man-made substances has proven not
to be the correct solution. When waste is bioremediated it can be recycled, or at least
stored in such a way that it causes no additional damage to the environment. This helps
prevent additional pollution from waste, or the product that treats it. Bioremediation is
one of the most cost effective and safe solutions we have now to manage polluted waste.

8
BIOFILMS
Biofilms may be defined as
assembly of microorganisms
comprising of different or
same microbial species,
attached to a biological or an inert surface and encased in a self-synthesized matrix.
Biofilms are not just microbial slime layers but biological systems. Microorganisms
organize themselves into coordinated functional community. The biofilm microbes can
share nutrients and are protected from harmful influences in the environment such as
desiccation, antibiotics and a host’s immune response. Biofilms have a three-dimensional
structure and they represent a community lifestyle for microorganisms. This is why they
are also described as “cities
for microbes”. Figure 2: Mycoplasma pneumonae biofilm on glass, 50 degree tilt,
20K magnification, by scanning electron microscopy (electron
Biofilms can form on both
micrograph scanned by Steven Distelhorst, Miami University)
living and non-living
surfaces. They can attach to
various surfaces such as a tooth or a rock and may include a single species or a diverse
group of microorganisms. A biofilm usually begins to form when a free-swimming
microorganism attaches to a surface.

FORMATION OF BIOFILMS

9
Initially, the first microorganism of a biofilm sticks to to a surface through weak Van der
Waals forces and hydrophobic properties. If this microorganism is not instantaneously
detached from the surface, it can anchor itself more enduringly using cell adhesion
structures such as pili.

Figure 3: Steps involved in biofilm formation

The hydrophobic nature of the microorganism also affects the formation of biofilm. A
microorganism which is highly hydrophobic will have reduced repulsion from the
substratum while a hydrophilic microorganism will be repulsed by the substratum.
Some microorganism species are not able to attach to a surface on their own due to
inadequate motility but are instead able to adhere themselves to the earlier microbe
colonists which are already attached to the surface. Non-motile microorganisms cannot
identify surfaces or combine together as easily as motile microorganisms.
Throughout surface colonization, the microbial cells are able to communicate using
quorum sensing (QS) products which regulates further development and modification of
the biofilm. Once colonization has commenced, the biofilm grows by a combination of
recruitment and cell division.
In the final stage of biofilm formation, also known as dispersion, the biofilm is
established and it only changes in shape or size.
10
Biofilms go through vigorous changes during their conversion from free living organisms
to sessile biofilm cells. These changes include a significant increase in their resistance
towards biological, chemical and physical stress and the specific production of secondary
metabolites.

EXTRACELLULAR POLYMERIC SUBSTANCES (EPS)

Biofilms are an aggregate of bacteria, protozoa and fungi enclosed in a matrix consisting
of a mixture of polymeric substances, commonly referred to as extracellular polymeric
substances (EPS). The EPS component which encases the cells is secreted by the cells
itself. EPS is typically a polymeric conglomeration of extracellular polysaccharides,
proteins, lipids and DNA. This matrix is so strong that under certain specific conditions,
biofilms can be fossilized.
EPS varies in composition between
species to species and this
composition is dependent on the
surface on which the biofilm is
attached, growth conditions and
environmental stress, among others.
Bacterial biofilms and EPS
production can undergo changes in
composition and structure based
upon the environmental conditions.
EPS is usually very strongly hydrated with internal water channels, however,
hydrophobic EPS also occur (e.g. EPS containing cellulose which is produced by a range
of microorganisms). The water channels present within some biofilms help in the
distribution of nutrients and signalling molecules.
This matrix also facilitates the
Figure 4: Bacillus subtilis cells grown on nutritive agar
communication among the cells
plates on (a) complex medium (LB) and (b) biofilm-
encased within it with the help of promoting minimal medium.
biochemical signals as well as gene
exchange. The EPS matrix also acts
as an external digestive system by trapping extracellular enzymes and keeping them in
close proximity.
EPS supports the high density of microbial biomass which facilitates the mineralization
process by sustaining ideal pH conditions, redox potential in the vicinity of the cells and

11
localized solute concentration. This is possible because of the controlled circulation of
fluids within the biofilm and its unique architecture.
The phenomenon of mass transport within this dense EPS is affected by its structure
which is determined by the local availability of the substrates. Solute transport in biofilms
is driven by two factors:
1. Convective transport within pores and water channels: The transmission of
heat due to bulk movement of molecules in fluids such as gases and liquids is
known as convection. It takes place through diffusion, advection or both.
2. Diffusion in the denser aggregates of cells: The movement of molecules from an
area of high concentration to an area of low concentration.
The microorganisms living inside a biofilm usually have significantly different properties
from the free floating microorganism of the same kind. The free living form of bacteria
inside a biofilms are known as planktonic cells. These are single-cells that may float or
swim in a liquid medium. The microbial cells growing inside a biofilms are
physiologically distinct from their planktonic cells as the dense and sheltered
environment of the biofilm allows them to communicate and cooperate in various ways.

12
BIOFILMS IN BIOREMEDIATION
For the past century, microbial communities have been utilized to neutralize, degrade and
mineralize many synthetic compounds in waste-water activated sludge. Even though
indigenous free-living planktonic microbial populations are capable of metabolising
persistent toxins and oxidizing heavy metal contaminants, their low abundance and
activity coupled with a lack of contact to the contaminants and limitations in available
nutrients in the environment inhibit these processes from happening at a sufficiently rapid
rate. Also, their survival in such a polluted environment is less likely due to lesser
protection, low metabolic activity and low bioavailability of the pollutant in the liquid
phase.
On the other hand, anchored and sessile microorganisms situated inside a biofilm
community afford structure and protection because of their development in a self-
produced and complex polymeric matrix. The ability of microbes to assemble in these
sessile biofilms structures permits for many benefits compared to their planktonic forms,
such as the ability to communicate and exchange genetic material, protection from the
surrounding environment, nutrient availability (from the environment and each other) and
persistence in different metabolic states. In addition, there is a genetic diversification in a
single species biofilm community and biofilms can also harbour diverse species of both
aerobic and anaerobic microorganisms, permitting them to metabolically supplement
each other and endure in the presence of varying nutrients. This metabolic range,
durability and persistence makes them very promising agents for bioremediation.
The array of features in a biofilm community exemplifies a sophisticated and sturdy
mechanism similar to different types of cells functioning as an organ. Encased within the
EPS, the biofilm structure grows and incorporates water channels which allows the
transport of nutrients and electron acceptor such as oxygen or other more reduced
molecular compounds. Additionally, the EPS contains surfactants which facilitate in
dissolving or emulsifying hydrophobic or other recalcitrant substrates that would

13
otherwise be inaccessible. These properties of biofilms has been well recorded in
biofilms found in diverse geographic regions such as streambeds, tidal flats, corroded
pipes and even sites of infection.
When biofilms are applied for bioremediation and biotransformation purposes, their
tolerance towards toxic and hazardous chemicals is of particular interest. This is because
the polluting chemicals can be present in such a high concentration in the environment
that it could be hazardous for the survival for their planktonic counter-parts. Biofilms
have a major advantage of increased tolerance towards environmental stress such as
decrease in nutrition, predation, exposure to chemicals (e.g. antibiotics or pollutants in
high concentration), change in pH, change in temperature etc., over free-floating
microorganisms. This high tolerance doesn’t only occur in the environment but can also
be observed in the biofilms in human body. The antibiotic concentration required to kill
free-living bacteria is often 100 to 1000 times lesser than that required for biofilms.

Table 1: Major properties of biofilms helpful in bioremediation and the characteristics associated with it.

PROPERTY
Tolerance towards environmental
stress (e.g. toxic chemicals, pH
change, predation etc.)
Communication (quorum sensing)
Exchange of genetic material
Metabolic diversity and symbiosis
Redox and electron acceptor
diversity
Varying growth rates in the biofilm
Porous physical structure with water
channels
Surfactants
Micro-colony and gradient
formation
CHARACTERISTIC
Extensive genetic diversification of biofilm
microorganisms.
Critical cell density for biofilm formation
Horizontal transfer of genetic material between
species, DNA sharing
Utilizing waste products and accumulated products
from the environment or other organisms
Different metabolic functions with respect to
electron-acceptor reduction
Inducing biofilm persistence due to different
metabolic states in the biofilm
Allow for transport of nutrients, electron acceptors,
and waste products
Aid in solubilizing hydrophobic or recalcitrant
substrates
Redox potential and nutrient cycling because of
aerobic and anaerobic processes

In the environment, particularly in soils and sediments, native biofilms constantly

perform bioremediation which is a part of global self-purification system and a part of
global nutrient cycling process. However, it is important to influence the kinetics of

14
biodegradation of organic contaminants and the immobilization of toxic metals to
improve the results and obtain the best outcome.
The significance of supporting biofilm growth to improve detoxification as well as the
longevity of microbial diversity and abundance has just recently been actively
appreciated and thus applied.
The characterization and isolation of bacteria like those of phylum Chloroflexi
(Dehalococcoides, Dehalobium chlorocoercia and o-17) capable of halorespiration (the
use of halogenated molecular compounds as terminal electron acceptor in anaerobic
respiration) has shed light onto the potential use of microbial processes for the
degradation of halogenated organic compounds. Furthermore, members of the genus
Dehalococcoids have been acknowledged as valuable inhabitants of a biofilm reactor
community for reductive dechlorination of trichloroethene.
Microbial communities can not only be utilized for bioremediation of organic pollutants
but they can also be applied towards metals. Passive oxidation of iron and arsenic by
biofilms was effective at gold-quarts mining sites, selenium has been reduced and
consequently concentrated via biofilms on tubes containing nutrients and in coal mine
drainage regions, biofilm enzymes have been engaged as biomarkers for stream water
quality. These examples demonstrate the diverse potential of biofilm-mediated
bioremediation and pollution monitoring set-ups.

15
BIOFILMS AND ORGANIC POLLUTANTS
Some of the most widely spread and most persistent pollutant found in the environment
today are organic compounds which are created for industrial applications. These are
known as persistent organic compounds or POPs and they include polychlorinated
biphenyl (PCBs), polychlorinated ethenes (PCEs), polycyclic aromatic hydrocarbons
(PAHs), polychlorinated dibenzo-p-dioxins and difurans (PCDD/Fs), etc. Most POPs are
hydrophobic in nature which leads them to accumulate in the food chain to toxic levels
and act as carcinogens and endocrine disruptors.
In addition to these contaminants, other xenobiotic substances such as pharmaceutical
and personal care products (PPCPs), fertilizers, herbicides and pesticides have also been
produced and dispersed in substantial quantities, polluting water and sediments world-
wide.
There are microorganisms that exist in the environment which have the capabilities to
metabolize and mineralize many of these xenobiotic substances. Bioremediation process
involves actively taking advantage of these naturally existing microbial biofilms to
engineer systems that can decontaminate and recycle these pollutants in the environment
more effectively.

1) POLYCYCLIC AROMATIC HYDROCARBONS (PAHs):

16
PAHs are consumed by benthic organisms and they make their way up on the food chain,
bioaccumulating in the fatty tissue of organisms with potential carcinogenic and
mutagenic effects.
PAHs have low solubility which leads to their low bioavailability in soil and sediments.
This poses to be a large obstacle, slowing down the process of bioremediation.
Introduction of surfactants could be a solution to this problem as it causes pseudo-
solubilisation of the insoluble PAH crystals but studies have shown that this pseudo-
solubilisation does not directly enhance the bioavailability of the PAHs.
Biofilm formation is the principle Figure 5: Polycyclic aromatic hydrocarbons: structures of
mechanism by microorganisms to some examples.
overcome mass transfer limitation
for insoluble crystallized PAHs. Studies have shown that mixed biofilms formed with
activated sludge inoculum reaches a higher biodegradation effectiveness on mixed PAH
substrate than if it is combined with a surfactant.
It is the co-metabolic mechanism of biofilms that enables the biodegradation of multiple
PAHs simultaneously. There are hundreds of PAH compounds that exist and a mixture of
these compounds is often found in the polluted environment, so biofilms are very
important when it comes to biodegradation and bioremediation of PAHs.

2) POLYCHLORINATED ETHENES (PCEs):

Chlorinated ethenes are another class of POPs that often threaten drinking water sources.
Dehalococcoides are a group of bacteria that are the only known microorganisms which
reduce PCE by specializing in
reductive dechlorination.

17
In a study by Chung et al (2008), hydrogen based denitrifying membrane biofilm reactor
(MBfR) was used to reductively dechlorinate TCEs. The TCE acted as an enrichment for
the Dehalococcoides and it was reported
that the bacteria was naturally present at
the beginning and end of the study.

3) POLYCHLORINATED BIPHENYLS (PCB):

PCBs are highly toxic POPs which were banned thirty years ago. Despite their ban, PCBs
can still be found in sediments, soil and even in air all around the world and the
contaminated sites are mainly treated with dredging and capping.
Figure 6: Reductive dechlorination of polychlorinated ethenes by catabolic organohalogen respirations.
Two physiologically distinguishable groups of bacteria mediate this process.

Figure 7: Structure of Polychlorinated Biphenyls

However, a group of microorganisms
called the dechlorinating Chloroflexi (e.g. Dehalococcoides Löffle, D.
lykanthroporepellens, D. chlorocoercia) has been found to have the ability to reductively
dechlorinate highly chlorinated PCBs under anaerobic conditions, therefore leaving less

chlorinated structures which are available for biodegradations aerobically.

An example of this is the anaerobic bacteria D. chlorocoercia DF1. Bioaugmentation of
sediments containing high concentration of PCBs by this bacteria affects doubly flanked
chlorines as well as it also stimulates indigenous dechlorinating microbial communities to
dechlorinate other PCBs. In the presence of a mature biofilm of the same bacterial

18
community, there is significant
microbial degradation of penta-
chlorobiphenyls on PCB oil
droplets.
Mature biofilms of
dehalogenating bacterial are
developed on GAC (granular
activated surface) which does not
affect the adsorption properties o
PCB or the formation of biofilms.
These are then delivered to the
sediments through water
columns. This set-up has greatly benefit the field of PCB bioremediation.

4) NITROAROMATIC COMPOUNDS
Nitroaromatic substances, like dinitrocresol and dinitrotoluene, are another class of
xenobiotic compounds which are resistant to biodegradation. However, bacteria that are
able to use several industrial nitroaromatic compounds, such as nitrophenols,
nitrobenzene and chloronitrobenzebe as nitrogen, carbon and energy sources have been
isolated.
The common pathway used to metabolize nitroaromatic substances is comparable to the
oxidative pathways used for aromatic acids and hydrocarbon metabolism, but with certain
suitable alterations to accommodate the nitro groups. Initial substrate are first converted
to substituted phenols, quinones or catechols by the bacteria. These are then metabolized
to intermediates of tricarboxylic acids (TCA) cycle.
In a study by Kumar et al, toluene was reported to be biodegraded using Burkholderia
vietnamiensis G4 in a laboratory-scale gas phase biofilm membrane reactor. There have
been reports of degradation of dinitrotluene in a mixed culture fluidized bed biofilm
reactor and degradation of toluene in hollow fibre membrane biofilter reactor and
continuously fed biodrum using
several aerobic and anaerobic
bacteria from secondary sludge
wastewater.

5) PHARMACEUTICAL & PERSONAL CARE PRODUCTS (PPCPs):

Figure 8:19Nitroaromatic compounds: structures of some examples.

In addition to the aforementioned polluting substances, there are various other xenobiotic
compounds that fall under the class of PPCPs. Many of these are not correctly regulated
in terms of wastewater (domestic and/or industrial) and are therefore emitted into the
environment as a part of effluent.
Triclosan, a commonly prescribed and industrially used antimicrobial is a particular
concern in this field. This chemical compound can be photo-transformed into dioxin
congeners in water columns. Triclosan being an antimicrobial not only harms the biofilm
communities that degrade dioxin, but it also turns into dioxin resulting in replenishment
of dioxin in nature. The biodegradation of this compound in conventional waste water
treatment facilities is inadequate, which results in the contamination of agricultural land
from the recycled sludge fertilizer. There is very little known about the microbial
communities responsible for the
Figure 9: Structure of Triclosan
trisclosan degradation but hopefully
further study within this filed will
improve the wastewater treatment process.
In terms of other PPCPs in the waste water, the biofilm reactors perform better than
activated sludge in the terms of removing several PPCPs (like diclofenac, trimethoprim,
metoprolol and gemfibrozil).

LIMITATIONS OF
BIOFILMS
Even though bioremediation processes
using biofilms and biofilm reactors have
many advantages over there traditional
remediation techniques, these are, quite
evidently, not highly favoured. This lack
of success is due to the following reasons:
1. Very few microorganisms present and/or have been identified to date which can be
utilized in these processes.
2. There is a lack of access to the pollutants for microorganisms due to adsorption.
3. Availability of electron donors/acceptors, water activity and co-metabolism act as
activity limiting factors.
Often, the challenges of biodegrading insoluble crystallized persistent organic polymers
(POPs) are the lack of indigenous microbes capable of performing bottleneck processes.

20
Only a limited number of microbial species are known to perform these processes and
their in-situ abundance is low. The bioavailability of hydrophobic compounds is another
limitation.
Another limitation of microbial bioremediation is reliability. Not all pollutants are
amenable to biodegradation by microorganisms or biofilms, particularly the man-made
synthetic recalcitrant substances such as plastics and certain halogenated aromatic
compounds. Also, during certain microbial degradation processes, some metabolic toxic
products could be generated. Moreover, application of some bioremediation in situ
solution might cause broader environmental problems, which has been the case for some
bioremediation strategies. Resuspension of sediment and thus the pollutant present in the
sediment increased the aqueous concentration to harmful levels for the residing microbial
population.
Bioremediation is also, relatively slower than the chemical treatment of pollutants for
degradation. Adequate technical expertise and interdisciplinary approach from different
fields such as environmental microbiology, civil engineering, soil sciences etc. are also
required for successful performance of biofilm mediated bioremediation.

CONCLUSION
Humans are capable of producing an enormous amount of pollutants is a well-known
fact. Remedying these pollutants with unnatural or man-made substances has proven not
to be the correct solution. When waste is bioremediated it can be recycled, or at least
stored in such a way that it causes no additional damage to the environment. This helps
prevent additional pollution from waste, or the product that treats it. Bioremediation is
one of the most cost effective and safe solutions we have now to manage polluted waste.

Biofilm bioremediation systems are continuously drawing more attention for research.
Biofilms have the potential to degrade xenobiotic substances into more manageable
forms. These systems are often more cost effective than conventional remediation efforts
such as dredging, capping incineration or extraction given that the end product is non-
polluting and these strategies might also cause smaller environmental foot prints.

21
However, biofilm mediated bioremediation also poses certain limitations. Further
research needs to be done before biofilm can be adapted into bioremediation processes up
to their full potential. Further study of biofilm communities would enable the
development of better methods for the bioremediation of polluted sites and wastewater.

REFERENCES
1. Applications of biofilms in bioremediation and biotransformation of persistent
organic pollutants, pharmaceuticals/personal care products, and heavy metals,
Sarah J. Edwards & Birthe V. Kjellerup, 2013
2. Application of Biofilms on Remediation of Pollutants – An Overview, Nilanjana
Das, Lakshmi V. Geetanjali Basak, Jaseetha Abdul Salam, Evy Alice Abigail M,
2012

3. Biofilm mediated decontamination of pollutants from the environment, Arindam

Mitra and Suman Mukhopadhyay, 2016

22
4. Biofilm-Mediated Enhanced Crude Oil Degradation by Newly Isolated
Pseudomonas Species, Debdeep Dasgupta, Ritabrata Ghosh, and Tapas K.
Sengupta, 2013

5. Biofilm-Mediated Heavy Metals Bioremediation in PGPR Pseudomonas, Amina

Meliani and Ahmed Bensoltane, 2016
6. Bioreduction of trichloroethene using a hydrogen‐based membrane biofilm
reactor. Environ Sci Technol, Chung J., Krajmalnik‐Brown R., Rittmann
B.E., 2008

7. Bioremediation, Environmental Enquiry, Cornell University and Penn State

University

8. Indoor Air Pollution: A Public Health Perspective, Spengler, John D, Sexton K.

A., 1983

9. Nitroaromatic Compounds: from Synthesis to Biodegradation, Kou-San Ju,

Rebecca E. Parales, 2010

10. Sequential Reductive Dechlorination of meta-Chlorinated Polychlorinated

Biphenyl Congeners in Sediment Microcosms by Two
Different ChloroflexiPhylotypes, Sonja K. Fagervold, Joy E. M. Watts, Harold D.
May, Kevin R. Sowers, 2005

11. Using Biofilms for the Detection and Bioremediation of Arsenic, Faculty of
Worchester Polytechnic Institute, Kayla DeSanty and Mitra Marvasti-Sitterly

23