Bio degradation of textile dyes in fluidized bed bioreactor

Abstract
The bio degradation of toxic textile dyes with fluidized bed bio reactor has been
studied in this review. Azo compounds constitute the largest and the most diverse group of
synthetic dyes and are widely used in a number of industries such as textile, food,
cosmetics and paper printing. They are generally recalcitrant to biodegradation due to their
xenobiotic nature. However microorganisms, being highly versatile, have developed
enzyme systems for the decolorization and mineralization of textile dyes under certain
environmental conditions. Reductive cleavage of azo bond, leading to the formation of
aromatic amines, is the initial reaction during the bacterial metabolism of azo dyes. Dye
decolorization by several mixed and pure bacterial cultures have been studied. Only few
aerobic bacterial strains that can utilize textile dyes as growth substrates have been isolated.
These organisms generally have a narrow substrate range. This review is focused on the
bacterial decolorization of azo dyes and mineralization of aromatic amines, as well as the
application of these processes for the treatment of textile-dye-containing wastewaters in
fluidized bed bioreactor. A different micro organism which has the tendency to decolorize
the dye on the basis of physio chemical properties like temperature, c.o.d, b.o.d has also
been studied for better understanding of the application point of view.
Keywords: aerobic/anaerobic bioreactor, degradation, dyes, decolourization,
fluidized bed bioreactor, textile effluent

Introduction:

Environmental pollution is an emerging threat and of great

concern

in todays context pertaining to its effect on the ecosystem. Water pollution is

one of the greatest concerns now a day. Industrial effluents often contain various
toxic metals, harmful gases, and several organic and inorganic compounds.
The worldwide rise in population and the industrialization during the last
few decades have resulted in ecological unbalance and degradation of the natural resources.
One of the most essential natural resources, which have been the worst

victim of

population explosion and growing industrialization, is water. Huge quantity of wastewater

generated from human settlement and industrial sectors accompany the disposal system
either as municipal wastewater or industrial wastewater.
Over 5 million chemical substances produced by industries have been
identified and about 12000 of these are marketed which amount to around half of the total
production. Due to discharge of toxic effluents long-term consequence of exposure can
cause cancer, delayed nervous damage, malformation in urban children, mutagenic
changes, neurological disorders etc. Thus it is imperative to purify and recycle wastewater
in view of reduced Availability and deteriorating water quality.
Textile industries are among the most polluting industries in
terms of the volume and the complexity of treatment of its efuents discharge. Wastewaters
generated by textile industries are known to contain large amounts of toxic aromatic
compounds, especially azo dyes. It is well known that some azo dyes and the degradation
products, such as aromatic amines, are highly carcino genic. Textile mill efuents are also
characterized by high levels of color caused by residual dyes. The colored wastewater
released into the ecosystem is also a dramatic source of aesthetic pollution and perturbation
in the aquatic life. Moreover, textile wastewaters are known to exhibit various pHs (either
alkaline or acidic, depending on the process used), hot temperature, high biological and
chemical (COD) oxygen demands and high concentrations of suspended solids.
Conventional methods for dealing with textile wastewater consist of various combinations

of biological, physical and chemical methods. Adsorption and precipitation processes are
very time- consuming and costly with low efciency. Chemical degradation by oxidative
agents such as chlorine constitutes the most relevant and effective method, but it may
produce some very toxic secondary products, such as organ chlorine compounds. Photo
oxidation by UV/H2O2 or UV/TiO2 needs additional chemicals and, therefore, may also
cause a secondary pollution. Although biodegradation process is cheaper than other
methods, it is less effective because of the toxicity of the dyes that induces an inhibiting
effect on bacterial development.
Textile effluents can seep into the aquifer and pollute the underground
water, or where it is discharged without proper treatment into water bodies, the pollutants
cannot be confined within specific boundaries. They can therefore affect aquatic life in
enormous ways. Metal effluents can have ecological impacts on water bodies leading to
increased nutrient load especially if they are essential metals. These metals in effluent may
increase fertility of the sediment and water column and consequently lead to
euthrophication. Dyes in water bodies also affect photosynthetic organisms and
consequently impact negatively on the food chain. The reason for this global effect also lies
in the weather systems and the biogeochemical cycling of elements, which aid in rapid
dispersal of the pollutants
Azo dyes are synthetic organic compounds characterized by the
presence of one or more azo bonds (N=N) in association with one or more aromatic
systems. Azo dyes account for the majority of all textile dyestuffs produced and are
currently the most commonly used synthetic dyes in textile, food, paper-making, and
cosmetic industries. Release of azo dyes into the environment from the effluents of dye
utilizing industries has become a major concern in wastewater treatment because azo dyes
may be mutagens or carcinogens.

The most common Bio-reactors are (1) Aerated lagoon, (2) Oxidation Ditch, (3)
Activated
Oxidation

sludge

system,

(4) Anaerobic

digestion

system,

(5)

pond, (6) Trickling filters, (7) Rotating discs biological reactors, (8)

Basket type

bioreactors, (9) Hollow fiber membrane bioreactor, and (10) Fluidized bed

bioreactors.
Aerobic processes have several advantages, including a large range of
wastewater that can be treated, high degree of BOD removal, acceptability of toxic
conditions, simultaneous nitrogen and phosphorous removal, better chlorinated organic
contaminants degradation, low solids retention time, and feasible small plants.
BIOLOGICAL TREATMENT

- Treatment Methods

The objectives of biological treatment of wastewater are to remove

the non-settleable colloidal solids and to degrade other organic matter. For industrial
wastewater, the objective is to remove or reduce the concentration of organic and inorganic
compounds. Because many of these compounds are toxic to microorganisms, pretreatment
may be required. With proper analysis and environmental control, almost all wastewaters
can be treated biologically. The removal of carbonaceous BOD is accomplished
biologically using a variety of microorganisms, mainly bacteria. The microorganisms are
used to convert the organic matter (colloidal and dissolved) into various gases and into cell
tissue. Unless the cell tissue that is produced from the organic matter is removed from this
solution, treatment has not been accomplished. The only treatment that has been achieved
is the conversion of a portion of the organics by bacteria to gaseous end products. There are
three major groups of biological processes:

Aerobic.

Anaerobic.

Combination of aerobic and anaerobic.

Considerable research efforts have been devoted to optimizing color

removal from effluents. Several physical and chemical treatment methods have been then
suggested but have not been widely applied because of the high cost and secondary

pollution that can be generated by the excessive use of chemicals. Microbial decolorization
of dye wastewater has been proposed as less expensive environmentally intrusive
alternative. In the natural environment, azo dyes can be transformed or degraded by a
variety of microorganisms, including aerobic and anaerobic bacteria and fungi. Bacterial
degradation of azo dyes is often initiated by an enzymatic biotransformation step that
involves cleavage of azo linkages with the aid of azo reductase using reduced coenzyme as
the electron donor, whereas fungal degradation of azo dyes was reported to be catalyzed by
extra cellular enzymes, ligninolytic peroxidases. Peroxides oxidize the phenolic ring of the
dyes to produce a carbonium ion on the carbon bearing the azo linkage. Water attacks the
carbonium ion to cause hydrolytic cleavage of the azo linkage. The use of fungi is a matter
of great concern as these organisms are able to degrade a wide variety of recalcitrant
organo pollutants, including various types of dyes The major enzymes associated with
lignin degradation are laccase, lignin peroxidase (LiP), and manganese peroxidase (MnP).
Some white rot fungi produce all three enzymes whereas others produce only one or two of
them.
Immobilization of living microorganisms has been described by several
investigators to be useful in biological wastewater treatment. It is widely known that
Immobilized cells offer a lot of advantages: reusability of the same biocatalyst, control of
reactions, and the non contamination of products. The purpose of this study was to
investigate the potential of micro organism to decolorize azo dyes like [orange G (OG),
trypan blue (TB), azorubine (AR), and methyl red (MR)] in fluidized bed bioreactor. The
effect of various physicochemical factors such as dye concentration, pH, and temperature
on the azo dyes decolorization and b.o.d, c.o.d were analyzed.

In the textile industries, the efuents are principally treated by biological

processes together with physico-chemical processes. Biological processes provide a low

cost and efficient alternative for simultaneous color and organic matter removal. The
biological processes (anaerobic and aerobic) can be divided into two systems according to
the nature of the biomass: suspended biomass systems (most manufacturing units)
and adhered biomass systems (biolm). Biolm is composed of cells, extracellular
polymers and organic and inorganic materials adhered to the solid surface, forming a thick,
voluminous layer. The xing of the microorganisms to the surface is the result of physical,
chemical and biological phenomena, the main factors which affect the formation and
maintenance of the biolm being: the support characteristics, the species of
microorganisms, the liquid phase and the uid dynamics. The development of the biolm is
determined by the balance between the growth and detachment of the cells (Drury et al.,
1993). This detachment is a function of friction forces between particles and shear stresses
caused by the displacement of liquid and gas (Chang et al., 1991). The biolm resistance is
a function of the extracellular polymer production.. (Ulson de Souza et al) The internal
structure of the biolm is still little known An understanding of the biolm structure, the
biodegradation kinetics occurring within the biolm and the impact of the biolm structure
on this kinetics is of fundamental importance for the optimization of the design and
operation of bioreactors with biolm (Bishop, 1997; Zhan et al., 2006). An important
parameter is the biolm diffusion which is inuenced by lm density, age, thickness, and
porosity, along with the presence of lamentous organisms, microbial speciation and
electrostatic interactions (Characklis and Marshall,1990).
Immobilized cells on the surface offer some advantages over cultures in
suspension such as: a greater variation in population; less sensitivity to environmental
variationstemperature, pH, toxic substances; higher growth rate and faster utilization of
the substrate in relation to free biomass. This is attributed to physiological modications
which the xed cells undergo, favoring changes in the cellular environment, due to
the increase in the local concentration of nutrients and enzymes, or the selective effect of
the extracellular polymeric matrix in relation to inhibitory or toxic substances
(Bishop,1997). One type of bioreactor with xed biomass and mobile supports is the
uidized bed reactor, in which particles covered in biolm are uidized by the recirculation
of liquid. These bioreactors are employed in aerobic processes (Lazarova and Manem,

1994), anaerobic processes (La Motta and Cascante, 1996) and denitrication processes
(Coelhoso et al., 1992; Green et al., 1995).
Fluidized bioreactors offer advantages over conventional processes
such as: they achieve a high concentration of biomass; a higher organic load can be
applied; hydraulic retention time is reduced; sludge production is lower; installations are
more compact; area of mass transfer between phases is large; and beds do not clog. On the
other hand there are some problems which inhibit their applicability on a large industrial
scale such as: control of the bed expansion, thickness of the biolm and oxygen distribution
system (Lazarova and Manem, 1994). Many support media are used: coal, sand, and
polymeric materials (PVC, polystyrene, OSBGoptimum support for growth, French
patent no. 8703611, March 1987) (Bergamasco and Tavares, 1998; Tavares et al., 1995).

Characteristics of aerobic bioreactors.

A large range of waste water can be treated. Purification and resettling
required. Can handle low to high CODs. Suitable for both cold and warm effluent.
Acceptable to toxic presence of toxic materials to certain extent. Neutralization of alkaline
wastewater required. Operated in continuous mode with less stability and control.
High oxygen requirement. Degree of BOD removal is also high. Simultaneous nitrogen
and phosphorous(nutrients) removal is possible. Posses high degradation rate to
Chlorinated organic contaminants. When carrier material is used leads to clogging danger.
Volumetric loading rates and solids retention time is low. Maintenance required for aeration
systems, sludge treatment. Has odour problems if open systems used. Sludge production is
high. Investment cost low

to medium. High

costs for aeration

(power),

nutrients,

sludge disposal. Small plants are possible.

Fluidized bed bioreactor in Wastewater Treatment

Over
immobilization

the

conventional

type

free-culture

bio-reactors

the

cell bioreactors like CSTR, PFR, fluidized bed, air lift type, etc. has the

following advantages like

continuous

reactor

operation

at

any

desired

liquid

throughput without risk of cell washout,

substrates,

higher

growth

rate

gives

protection

of cells from

high concentration

of

cells

toxic
in

the

reactor, easy cell-treated water separation, enhanced gas- liquid mass transfer rate,
plug flow operation by maintaining the immobilized cells as a stationary
fluidized bed bioreactors

are superior in performance due to immobilization

of cells on solid particles reduce

extremely

small,

lack

phase.The

the time of treatment, volume

of clogging

of reactor

is

of bio-mass and removal of phenol even at

lower concentrations .
A fluidized
treatment

bed

bioreactor

(FBB)

is capable

of achieving

in low retention time because of the high biomass concentration. FBB offers

distinct mechanical advantages, which allow small and high surface area media
to be used for biomass growth. Fluidization overcomes operating problems such
as

bed

clogging and the high-pressure drop, which would occur if small and high

surface area media were employed

in packed-bed

operation.

Rather than clog

with new biomass growth, the fluidized bed simply expands. Thus for a comparable
treatment efficiency, the required

bioreactor volume is greatly reduced. A further

advantage is the possible elimination of the secondary

be weighed

against the medium-biomass Separator.

clarifier, although

this must

Conventional FBB are

operated in two different ways. In a bioreactor with a heavy (matrix particle

density larger than that of liquid) biomass support (e.g. silica sand, coal), fluidization is
commonly

conducted

with

an

upward

co

current

flow

of

gas

and liquid

through a bed of particles. Under fluidization conditions, the bed is fluidized with an
upward flow of a liquid counter to the net gravitational force of the particles. Once
fluidized, each particle provides a large surface area for biofilm formation and growth. The
support media eventually become covered with biofilm and the vast available growth
surface afforded by the media results in a biomass concentration approximately an order of
magnitude greater than that maintained in a suspended growth system.
A practical problem, which occurs in the operation of an FBB, is the
excessive growth of biomass on support media. This can lead to the channeling of

bioparticles in fluidized beds since biomass loading can increase to such extent
that the bioparticles

began to be carried over from a bioreactor. The problem of over

expansion of fluidized bed due to biomass growth has generally been solved by the
removal of heavily biomass- laden particles from

bioreactor, followed by the addition

of biomass-free particles. However this solution complicates operation of a bioreactor and

introduces the need for additional
vibrating

screen

equipment external to the bioreactor, such as a

or an incinerator. The

nutrients

for

microbial

growth

are

transported first from bulk phase to the surface of the biofilm, and then transported to
the

inner

regions

of

the

biofilm

via diffusion. The limiting mass transport rate

controls the performance of the biofilm reactor.

Literature survey:
Literature survey is carried out to study

the different physio

chemical properties which affects the degradation process of toxic textile dye. The different
type of dyes and their percentage removal has also been studied. The type of reactor and
type of organism used can be compared and the performance has also been studied. T
Youssef Zeroual et al has been worked with Decolorization of Some Azo
Dyes by Immobilized Geotrichum sp. Biomass in Fluidized Bed Bioreactor It has been
shown that immobilized Geotrichum sp. biomass could effectively decolorize at high level
azo dyes in fluidized bed bioreactor. The obtained results reveal that this bioprocess
facilitates reproducible decolorization of azo dye with high yield. The use of Geotrichum
sp. entrapped in polyacrylamide gel in repeated batch fluidized bioreactor has proven not to
be interesting because the decolorization system lost more than 70% of its decolorization
efficiency at the end of the third cycle of treatment.
Sanjay Seshadri et al (1994 has studied The primary objective of this
research was to investigate the potential for removal of Azo dyes found in wastewater using
a two-stage anaerobic/aerobic reactor system. That

paper focusses on the removals

achieved in the first-stage anaerobic fluidized-bed reactor. It was found that Azo dye

transformation (probably by azo bond cleavage), for each of the four dyes tested, was
readily accomplished under anaerobic conditions; however, complete mineralization was
not achieved, necessitating a second-stage aerobic reactor.
Adnane Moutaouakkil et al 2003 has been shown that high-efficiency
decolorization of water contaminated by azo dyes with immobilized azo dye-degrading
bacteria in a fluidized bed bioreactor is feasible. The obtained results show that this
technology allows specific and reproducible decolorization of the azo dye MR with high
yield. The specific decolorization rate obtained by immobilizing Enterobacter
agglomerans in polyacrylamide is superior to those obtained by fixing this bacterial strain
on cooper beech or vermiculite. However, the entrapment of E. agglomerans in calcium
alginate gel offers the greatest purifying performance as well as the biggest stability of the
bioreactor. Moreover, the hydrodynamic behavior of the beads, in a fluidized bed
bioreactor, is very satisfactory. The high specific decolorization rate obtained and the
simplicity of the immobilization method mean that alginate would be a suitable
immobilization matrix for using bacterial strains to remove azo dyes from wastewater on an
industrial scale.
Sharma et al2004 did the experiments with Acid blue-15 a complex and
resonance-stabilized triphenylmethane (TPM) textile dye, resistant to transformation, was
decolorized/ degraded in an up-flow immobilized cell bioreactor. A consortium comprised
of isolates belonging to Bacillus sp., Alcaligenes sp. And Aeromonas sp. formed a
multispecies biofilm on refractory brick pieces used as support material. The TPM dye was
degraded to simple metabolic intermediates in the bioreactor with 94% decolorization. The
results of this study show that the developed up flow immobilized cell bioreactor can
enhance the ability of the microbial consortium to decolorize and degrade industrially
important textile dyes
D. Georgiou et al 2006 conducted experiment in Textile wastewater was
treated utilizing a bench-scale fluidized-bed loop reactor with porous carriers colonized by
anaerobic bacteria. Acetic acid solution (enriched with nutrients and trace elements) was

used both as a pH-regulator and as an external substrate addition for the growth of
methanogenic bacteria. The main aim of the treatment was decoloration of the textile
wastewater and transformation of the non-biodegradable azo-reactive dyes to the
degradable, under aerobic biological conditions, aromatic amines. The above process
method is very efficient since it is able to decolorize textile wastewater at very low
hydraulic residence times
Anjali Pandeyet al 2006 reported that Degradation of aromatic amines
depends on their chemical structure and the conditions. It is now known that simple
aromatic amines can be mineralized under methanogenic conditions. Sulfonated aromatic
amines, on the other hand, are resistant and require specialized aerobic microbial consortia
for their mineralization. This review is focused on the bacterial decolorization of azo dyes
and mineralization of aromatic amines, as well as the application of these processes for the
treatment of azo-dye-containing wastewaters.
Anjali Pandey et al found that the presence of dyes imparts an intense
color to effluents, which leads to environmental as well as aesthetic problems. The
treatment of azo-dye-containing wastewater

still presents a technical challenge. As

regulations are becoming even more stringent, there is urgent need for technically feasible
and cost-effective methods. The available literature seems to indicate that anaerobic aerobic
biological methods may be appropriate for the treatment of dye-containing wastewaters.
However, there is a still a need to assess the extent of mineralization of aromatic amines, as
many amines can undergo autoxidation, leading to the formation of soluble recalcitrant
polymers, which may be toxic.

Degradation of many amines, including SAA, requires the presence of specialized

cultures. SAA degraders have a very narrow substrate range. Hence there is a requirement
for developing microbial consortia that harbor genes for the rapid degradation of mixtures
of aromatic amines. Such cultures may have to be used for the bioaugmentation of aerobic
treatment units. Molecular biology techniques may also be used to improve the strains so

that rapid mineralization of aromatic amines can be achieved. Their use, however, requires
caution. It may also be necessary to combine AOP with biological processes to achieve the
required degree of treatment of dye-containing wastewaters so that regulatory standards can
be met
Souza et al 2008 study reveals that the application of aerobic processes with
biofilms is an excellent alternative for reducing COD in industrial textile effluents, without
the use of chemicals compounds, which are utilized in the traditional effluent treatment
processes, making this biofilm technology environmentally friendly

Micro organism

carrier

substrate

Reactor
type

reference

Mixed culture

Marble chips

Textile
effluent

Immobilized
bioreactor

Bella devassy
tony et
al(2009)

Direct red 28
Textile
effluent

bioreactor

Eltaief khelifi
et al (2009)

Indigo
Textile
effluent

3 phase f.b.b.r

Souza et
al(2007)

Textile
effluent

f.b.b.r

Jane yii wu et
al(2005)

Textile
effluent

Fluidized bio
reactor

Adnane
moutaouakkil
et al (2004)

b.vallismortis,pumilus
cerus bacillus sp.
Mixed culture
Clostridium spp.
Desulfovibrio spp.
Mixed culture

Mixed culture
pure culture

Glass cylinder
filled with
flocor
Small p.v.c
cylinders
Polyphorylated
pva gel in glass
beads
Calcium
alginate beads

Results and recommendations

Dyeing methods and its subsequent removal in a series operation

consisting a combination of unit operations reported above. And hence, in the present
study, emphasis is given to reduction of COD from the effluents generated due to the usage
of three different types of reactive dyeing methods.
D.k sharma et al 2004 study, they examined the decolorization and
degradation of acid blue-15, a commercially available TPM dye, by a bacterial consortium
immobilized on refractory brick pieces in an up-ow cell bioreactor. AB-15 has a more
complex structure than other well studied model TPM dyes, like crystal violet and
malachite green, etc., due to the presence of additional phenyl rings on the basic triphenyl
structure These substituents, although they give intense color and dye-fastness, result in a
resonance-stabilized structure recalcitrant to trans formation in the environment. This
results in biomagnication, leading to toxic responses in target populations. The individual
bacterial isolates showed less than 30% decolorization of 20 mg dye shake-ask studies,
whereas the consortium of these isolates resulted in 70% decolorization of AB-15.
Bella Devassy Tonyet al 2009 In their study, high-efciency
decolorization of azo dye Direct Red 28 was achieved by immobilization of azo dyedegrading mixed bacterial culture SKB-II comprised of ve bacterial isolates, B. pumillus,
B. cereus, B. megaterium, B. vallismortis and B. subtilis, in a packed bed bioreactor. 91 to
72%.
Mahidi harun et al 2008 has reported that
% Efficiency in removal, %
Method
COD Color
Two-phase anaerobic packed bed reactor
78
90
Aerobic/aerobic using sequencing batch reactors
76
94
Aerobic/aerobic sequential treatment system
90
85

Franciscon Elisangela et el 2009 A facultative Staphylococcus arlettae bacterium,

isolated from an activated sludge process in a textile industry, was able to successfully

decolourize four different azo dyes under microaerophilic conditions (decolourization

percentage >97%).
Sangyong kim et al 2003 studied COD Reduction and Decolorization
of Textile Effluent Using a Combined Process. This study showed the effectiveness
of biological pretreatment involving appropriate microorganisms and suitable support
media in a combined

process. The combined process consists of biological

pretreatment, chemical coagulation and electrochemical oxidation. COD and color

were reduced by 95.4% and 98.5% by the combined process, respectively.
Frank P. van der Zee According to the concept of combined anaerobicaerobic
treatment, azo dyes should be removed from the water phase by (anaerobic) reduction
followed by (aerobic) oxidation of the dyes constituent aromatic amines. The anaerobic
aerobic reactor studies reviewed in this paper show that a generally high extent of color
removal can be obtained, and several studies further more provide evidence for removal of
aromatic amines. Combined anaerobicaerobic treatment therefore holds promise as a
method to completely remove azo dyes from wastewater. However, the results of the
reactor studies reveal some possible limitations, both with respect to azo dye reduction and
with respect to the fate of aromatic amines.
The various reactors and micro organisms performance has been
tabulated above. rather than aerobic and anaerobic methods the combination of two
methods has also shows significant improvence in dye degradation, at the same time the
combination of two methods has also been developed now a day fox example fbbr with
Fenton reagent, electro chemical method, photo catalytic method, shows significant colour
removal.

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