ARTICLE IN PRESS

Wastewater treatment in molasses-based alcohol distilleries for COD

and color removal: A review
Y. Satyawali, M. Balakrishnan
TERI University, Darbari Seth Block, India Habitat Centre, Lodhi Road, New Delhi 110 003, India

Abstract

Molasses-based distilleries are one of the most polluting industries generating large volumes of high strength wastewater. Different
processes covering anaerobic, aerobic as well as physico-chemical methods have been employed to treat this effluent. Anaerobic
treatment is the most attractive primary treatment due to over 80% BOD removal combined with energy recovery in the form of biogas.
Further treatment to reduce residual organic load and color includes various: (i) biological methods employing different fungi, bacteria
and algae, and (ii) physico-chemical methods such as adsorption, coagulation/precipitation, oxidation and membrane filtration.
This work presents a review of the existing status and advances in biological and physico-chemical methods applied to the treatment of
molasses-based distillery wastewater. Both laboratory and pilot/industrial studies have been considered. Furthermore, limitations in the
existing processes have been summarized and potential areas for further investigations have been discussed.

Keywords: Decolorization; Distillery; Molasses; Spentwash

1. Introduction photosynthesis by blocking sunlight and is therefore

Ethanol manufacture from molasses generates large
volumes of high strength wastewater that is of serious
environmental concern. The effluent is characterized by
extremely high chemical oxygen demand (COD)
(80,000–100,000 mg/l) and biochemical oxygen demand
(BOD) (40,000–50,000 mg/l), apart from low pH, strong
odor and dark brown color (Central Pollution Control
Board (CPCB) 1994, 2003). In India, which is the second-
largest producer of ethanol in Asia with a projected annual
production of about 2300 million liters in 2006–07
(Subramanian et al., 2005), alcohol distilleries are rated
as one of the 17 most polluting industries. Apart from high
organic content, distillery wastewater also contains nu-
trients in the form of nitrogen (1660–4200 mg/l), phos-
phorus (225–3038 mg/l) and potassium (9600–17,475 mg/l)
(Mahimairaja and Bolan, 2004) that can lead to eutrophi-
cation of water bodies. Further, its dark color hinders
Corresponding author. Tel.: +91 11 2468 2100; fax: +91 11 2468 2144.
E-mail address: malinib@teri.res.in (M. Balakrishnan).
deleterious to aquatic life (FitzGibbon et al., 1998). Studies
on water quality of a river contaminated with distillery
effluent displayed high BOD values of 1600–21,000 mg/l
within a 8 km radius (Baruah et al., 1993). Adequate
treatment is therefore imperative before the effluent is
discharged. In addition to pollution, increasingly stringent
environmental regulations are forcing distilleries to im-
prove existing treatment and also explore alternative
methods of effluent management. For instance, Indian
distilleries were stipulated to achieve zero discharge of
spentwash to inland surface water by December 2005
(Uppal, 2004).
In an earlier review on this subject, Sheehan and
Greenfield (1980) discussed treatment options practiced
in the 1970s. More recently, Wilkie et al. (2000) have
examined characteristics and anaerobic treatment of
effluent obtained from different feedstock used for ethanol
manufacture. This review focuses on the advances in
molasses-based distillery wastewater treatment in the
last two decades and the emerging technologies in this
field.
 ARTICLE IN PRESS

2. Process description manufacturing process and handling and storage (God-

Alcohol manufacture in distilleries consists of four main
steps viz. feed preparation, fermentation, distillation and
packaging (Fig. 1).
2.1. Feed preparation
Ethanol can be produced from a wide range of feedstock.
These include sugar-based (cane and beet molasses, cane
juice), starch-based (corn, wheat, cassava, rice, barley) and
cellulosic (crop residues, sugarcane bagasse, wood, muni-
cipal solid wastes) materials. Details of sugar, starch and
lignocellulosic biomass-based feedstock and their pre-
treatment steps have been reviewed earlier (Wilkie et al.,
2000). In general, sugar-based feedstock containing readily
available fermentable sugars are preferred since starch and
cellulosic substrates involve an additional pre-treatment
step to convert starch into fermentable sugars. Thus, cane
juice is a commonly used substrate in Brazil (Tano and
Buzato, 2003) while Indian distilleries almost exclusively
use sugarcane molasses. Overall, nearly 61% of world
ethanol production is from sugar crops (Berg, 2004).
The composition of molasses varies with the variety of
cane, the agro climatic conditions of the region, sugar
Yeast Pre-fermenter
Diluted CO2
molasses Fermenter
bole, 2002). Table 1 summarizes the chemical composition
of beet and cane molasses. Molasses is diluted to about
20–25 brix (measurement of sugar concentration in a
solution) and its pH adjusted, if required, before fermenta-
tion. In India, about 90% of the molasses produced in cane
sugar manufacture is consumed in ethanol production
(Billore et al., 2001).
2.2. Fermentation
Yeast culture is prepared in the laboratory and
propagated in a series of fermenters, each about 10 times
larger than the previous one. The feed is inoculated with
about 10% by volume of yeast (Saccharomyces cerevisiae)
inoculum. This is an anaerobic process carried out under
controlled conditions of temperature and pH wherein
reducing sugars are broken down to ethyl alcohol and
carbon dioxide. The reaction is exothermic. To maintain
the temperature between 25 and 32 1C plate heat exchan-
gers are used; alternatively some units spray cooling water
on the fermenter walls. Fermentation can be carried out in
either batch or continuous mode (CPCB, 2003). Fermenta-
tion time for batch operation is typically 24–36 h with an
efficiency of about 95%. Continuous operation, involving
higher sugar concentration and an osmotolerant variety of
yeast, is faster (16–24 h fermentation time) but the
efficiency is marginally lower (T.R. Sreekrishnan, pers.
comm.). The resulting broth contains 6–8% alcohol. The
sludge (mainly yeast cells) is separated by settling and

Analyzer Table 1
column Composition of cane and beet molasses (Curtin, 1983; Chen and Chou,
1993; Godbole, 2002)
Spentwash
Property Cane molasses Beet molasses

Rectification Brix (%) 79.5 79.5

column 85–92b
Specific gravity 1.41 1.41
Spentlees 1.38–1.52a
Total solids (%) 75.0 77.0
75–88a
Alcohol
Total sugars (%) 46.0 48.0
44–60a
50–90b
Crude protein (%) 3.0 6.0
Blending & Dehydration 2.5–4.5b
maturation (molecular sieve) Total fat (%) 0.0 0.0
Total fiber (%) 0.0 0.0
Ash (%) 8.1 8.7
7–15b
Industrial Calcium (%) 0.8 0.2
Potable Power
alcohol alcohol Phosphorus (%) 0.08 0.03
alcohol
Potassium (%) 2.4 4.7
Sodium (%) 0.2 1.0
Chlorine (%) 1.4 0.9
Sulfur (%) 0.5 0.5
Bottling
a
Godbole (2002).
Fig. 1. Process description. b
Chen and Chou (1993).
 ARTICLE IN PRESS
discharged from the bottom, while the cell free fermenta-
tion broth is sent for distillation.
Apart from yeast, the bacterium Zymomonas mobilis has
also been investigated for ethanol production (Tao et al.,
2005). The organism follows a simple catabolic pathway
(Toma et al., 2003) and has advantages over S. cerevisiae
because of its higher sugar uptake rate, lower biomass
yields and higher ethanol production (Lin and Tanaka,
2006). Immobilization of Zymomonas cells has been tried
on support materials such as pectin because immobilized
whole cells retain higher microbial activity (Siva Kesava
et al., 1996). However, formation of byproducts such as
acetoin, glycerol, acetate, and lactate under anaerobic
conditions is a drawback associated with Z. mobilis.
During its growth on sucrose medium, Z. mobilis also
leads to formation of levan, a polymer of fructose units.
Further, like yeast, Zymomonas is unable to convert
complex carbohydrates like cellulose, hemicellulose and
starch to ethanol. To overcome these constraints, genetic
manipulation of Z. mobilis has been attempted by several
workers. For instance, the hydrolytic and isomerase genes
from recombinant Escherichia coli have been transferred to
Z. mobilis, resulting in utilization of xylose, mannose,
lactose and arabinose as the carbon source (Gunasekaran
and Raj, 1999). Earlier, Picataggio et al. (1996) reported
the development of genetically altered Zymomonas strain
to ferment pentoses and glucose, obtained by hydrolysis of
hemicellulose and cellulose, to produce ethanol. Here, Z.
mobilis was transformed with combination of E. coli genes
for xylose isomerase, xylulokinase, transaldolase, transke-
tolase, L-arabinose isomerase, L-ribulokinase, and L-ribu-
lose-5-phosphate 4-epimerase. In another work aimed at
xylose utilization combined with enhanced ethanol yield, a
strain derived from Z. mobilis ATCC31821 was developed
(Zhang, 2000). The strain comprised of exogenous genes
encoding xylose isomerase, xylulokinase, transaldolase and
transketolase.
2.3. Distillation
Distillation is a two-stage process and is typically carried
out in a series of bubble cap fractionating columns. The
first stage consists of the analyzer column and is followed
by rectification columns. The cell free fermentation broth
Table 2
Sample quantities and characteristics of wastewater streams generated in an Indian distillery (S. Majumdar, pers. comm.)
Parameter Specific wastewater Color
generation (kl/kl alcohol)
Spent wash 14.4 Dark brown
Fermenter cleaning 0.6 Yellow
Fermenter cooling 0.4 Colorless
Condenser cooling 2.88 Colorless
Floor wash 0.8 Colorless
Bottling plant 14 Hazy
Other 0.8 Pale yellow
(wash) is preheated to about 90 1C by heat exchange with
the effluent (‘‘spentwash’’) and then sent to the degasifying
section of the analyzer column. Here, the liquor is heated
by live steam and fractionated to give about 40–45%
alcohol. The bottom discharge from the analyzer column is
the spentwash. The alcohol vapors are led to the
rectification column where by reflux action, 96% alcohol
is tapped, cooled and collected. The condensed water from
this stage, known as ‘‘spentlees’’ is usually pumped back to
the analyzer column.
2.4. Packaging
Rectified spirit (96% ethanol by volume) is marketed
directly for the manufacture of chemicals such as acetic
acid, acetone, oxalic acid and absolute alcohol. Denatured
ethanol for industrial and laboratory use typically contains
60–95% ethanol as well as between 1% to 5% each of
methanol, isopropanol, methyl isobutyl ketone (MIBK),
ethyl acetate, etc. (Skerratt, 2004).
For beverages, the alcohol is matured and blended with
malt alcohol (for manufacture of whisky) and diluted to
requisite strength to obtain the desired type of liquor. This
is bottled appropriately in a bottling plant. Anhydrous
ethanol for fuel-blending applications (‘‘power alcohol’’)
requires concentration of the ethanol to 499.5 wt% purity.
The ethanol dehydration is typically done using molecular
sieves; however, pervaporation has also been employed in
Brazil and India for this purpose (Mitsui & Co., 2003).
3. Wastewater generation and characteristics
Table 2 lists the major wastewater streams generated at
different stages in the alcohol manufacturing process.
Table 3 summarizes the typical characteristics of spentwash
generated in Indian distilleries using sugarcane molasses.
Values for beet molasses-based effluent are given for
comparison. The main source of wastewater generation is
the distillation step wherein large volumes of dark brown
effluent (termed as spentwash, stillage, slop or vinasse) is
generated in the temperature range of 71–81 1C (Yeoh,
1997; Nandy et al., 2002; Patil et al., 2003). The
characteristics of the spentwash depend on the raw
material used (Mall and Kumar, 1997); also, it is estimated
pH Suspended solids BOD (mg/l) COD (mg/l)
(mg/l)
4.6 615 36,500 82,080
3.5 3000 4000 16,500
6.3 220 105 750
9.2 400 45 425
7.3 175 100 200
7.6 150 10 250
8.1 100 30 250
 ARTICLE IN PRESS

Table 3
Characteristics of spentwash generated from various feedstock (Pathade, 1999; Wilkie et al., 2000; Mahimairaja and Bolan, 2004)

Characteristics Feedstock

Cane molasses Beet molasses

Pathade (1999) Mahimairaja and Bolan (2004) Wilkie et al. (2000)

COD (mg/l) 65,000–130, 000 104,000–134,400 91,100

BOD (mg/l) 30,000–70,000 46,100–96,000 44,900
COD/BOD ratio 2.49 1.95
Total solids (mg/l) 30,000–100,000
Total suspended solids (mg/l) 350
Total dissolved solids (mg/l) 80,000 79,000–87,990
Total nitrogen (mg/l) 1000–2000 1660–4200 3569
Total phosphorus (mg/l) 800–1200 225–3038 163
Potassium (mg/l) 8000–12,000 9600–17,475 10,030
Sulfur as SO4 (mg/l) 2000–6000 3240–3425 3716
pH 3–5.4 3.9–4.3 5.35

that 88% of the molasses constituents end up as waste (Jain Spentwash

et al., 2002). Molasses spentwash has very high levels of
BOD, COD, COD/BOD ratio as well as high potassium,
phosphorus and sulfate content (Table 3). In addition, cane
molasses spentwash contains low molecular weight com-
pounds such as lactic acid, glycerol, ethanol and acetic acid
(Wilkie et al., 2000).
Cane molasses also contains around 2% of a dark brown
pigment called melanoidins that impart color to the
spentwash (Kalavathi et al., 2001). Melanoidins are low
and high molecular weight polymers formed as one of the
final products of Maillard reaction, which is a non-
enzymatic browning reaction resulting from the reaction
of reducing sugars and amino compounds (Martins and
van Boekel, 2004). This reaction proceeds effectively at
temperatures above 50 1C and pH 4–7. The structure of
melanoidins is still not well known (Rivero-Pérez et al.,
2002). Only 6–7% degradation of the melanoidins is
achieved in the conventional anaerobic–aerobic effluent
treatment process (Gonzalez et al., 2000). Due to their
antioxidant properties, melanoidins are toxic to many
microorganisms involved in wastewater treatment (Siria-
nuntapiboon et al., 2004a). Apart from melanoidins,
spentwash contains other colorants such as phenolics,
caramel and melanin. Phenolics are more pronounced in
cane molasses wastewater whereas melanin is significant in
beet molasses (Godshall, 1999).
4. Effluent treatment
Till the early 1970s, land disposal was practiced as one of
the main treatment options, since it was found to enhance
yield of certain crops. For example, in Brazil, vinasse
generated from sugarcane juice fermentation is mainly used
as a fertilizer due to its high nitrogen, phosphorus and
organic content. Its use is further reported to increase
sugarcane productivity; furthermore under controlled
conditions, the effluent is capable of replacing application
Biomethanation
Biocomposting Aerobic Solar Evaporation /
treatement drying incineration
Dilution Tertiary treatment
Surface water
discharge
Irrigation
Fig. 2. Spentwash treatment options.
of inorganic fertilizers (Cortez and Peréz, 1997; Rodrı́guez,
2000). However, for the high strength molasses-based
spentwash, the odor, putrefaction and unpleasant land-
scape due to unsystematic disposal are concerns in land
application. In addition, this option is subject to land
availability in the vicinity of the distillery; also, it is
essential that the disposal site be located in a low–medium
rainfall area (Sheehan and Greenfield, 1980). More recent
investigations have indicated that land disposal of distillery
effluent can lead to groundwater contamination (Joshi,
1999). Deep well disposal is another option but limited
underground storage and specific geological location limits
this alternative. Other disposal methods like evaporation of
spentwash to produce animal feed and incineration of
spentwash for potash recovery have also been practiced
(Sheehan and Greenfield, 1980; Wilkie et al., 2000).
Fig. 2 presents the options currently employed for
molasses spentwash treatment. The salient features of the
existing options as well as the advances in this field are
discussed in the following sections.
 ARTICLE IN PRESS
4.1. Biological treatment
4.1.1. Anaerobic process
The high organic content of molasses spentwash makes
anaerobic treatment attractive in comparison to direct
aerobic treatment. Therefore, biomethanation is the
primary treatment step and is often followed by two-stage
aerobic treatment before discharge into a water body or on
land for irrigation (Nandy et al., 2002). Aerobic treatment
alone is not feasible due to the high energy consumption
for aeration, cooling, etc. Moreover, 50% of the COD is
converted to sludge after aerobic treatment (Sennitt, 2005).
In contrast, anaerobic treatment converts over half of the
effluent COD into biogas (Wilkie et al., 2000). Anaerobic
treatment can be successfully operated at high organic
loading rates; also, the biogas thus generated can be
utilized for steam generation in the boilers thereby meeting
the energy demands of the unit (Nandy et al., 2002).
Further, low nutrient requirements and stabilized sludge
production are other associated benefits (Jiménez et al.,
2004).
The performance and treatment efficiency of anaerobic
process can be influenced both by inoculum source and
feed pre-treatment. In particular, thermal treatment of
wastewaters can result in rapid degradation of organic
matter leading to lower hydraulic residence time (HRT),
higher loading rate and BOD reduction. Moreover the
methane content and calorific value of biogas produced
from thermophilic systems was higher (Vlissidis and
Zouboulis, 1993). On the contrary, in shake flask studies
with an initial COD loading of 20,000 mg/l using biogas
plant sludge, Dhar et al. (1998a) observed a COD removal
of just 6.5% with thermally pre-treated spentwash whereas
untreated influent displayed 30.4% COD removal. Sig-
nificant improvement was observed using inoculum from
anaerobic lagoon with 27.2% COD reduction with
thermally pre-treated wastewater and 51% reduction with
untreated wastewater. Thermal pre-treatment changes the
biodegradability of wastewater; thus, it acts as an entirely
new feedstock for which the inoculum has to be
acclimatized afresh. Further, the temperature of thermal
pre-treatment is also important. After 150 d adaptation
period, wastewater treated at lower temperature (170 1C)
showed 66% COD reduction (Dhar et al., 1998b). This was
nearly twice the removal obtained with treatment at 230 1C.
Further, addition of micronutrients (iron, boron and
molybdenum) eliminated the long adaptation periods.
Anaerobic lagoons are the simplest option for the
anaerobic treatment of distillery spentwash. Subba Rao
(1972) reported that employing two anaerobic lagoons in
series resulted in final BOD levels up to 600 mg/l. However,
large area requirement, odor problem and chances of
ground water pollution restrict its usage (Pathade, 1999).
Though anaerobic lagoons are still employed in Indian
distilleries, high rate anaerobic reactors are more popular
(Lata et al., 2002). These reactors offer the advantage of
separating the hydraulic retention time (HRT) from solids
retention time (SRT) so that slow growing anaerobic
microorganisms can remain in the reactor independent of
wastewater flow. Table 4 summarizes the performance of
various anaerobic reactors covering both laboratory
studies and pilot/commercial scale operations for treatment
of molasses-based distillery wastewaters.
4.1.1.1. Suspended bed reactor. Upflow anaerobic sludge
blanket (UASB) reactor is the most popular high rate
digester that has been utilized for anaerobic treatment of
various types of industrial wastewaters (Akunna and
Clark, 2000; Syutsubo et al., 1997). Treatment by a UASB
reactor resulted in 75% COD removal in sugarcane
molasses spentwash and 90% COD reduction in whisky
pot ale (Goodwin and Stuart, 1994; Sánchez Riera et al.,
1985). However, dilution is required before treatment due
to the presence of some inhibitory substances such as sulfur
compounds, potassium and calcium ions and free hydrogen
ions left in solution after pH correction (Sánchez Riera
et al., 1985). Wolmarans and de Villiers (2002) have
reported a similar COD removal efficiency of greater than
90% over three seasons in a UASB plant treating distillery
wastewater.
Most of the practical UASB systems are operated under
mesophilic conditions; however, thermophilic operation
results in higher methanogenic activity. Mesophilically
grown sludge utilized in thermophilic UASB as a seeding
material leads to prompt start up and stable operation with
85% COD removal efficiency at a loading of 30 kg COD/
m3 d (Syutsubo et al., 1997). There are also reports on the
cultivation of thermophilic granular sludge for the seeding
of thermophilic UASB reactor. Wiegant et al. (1985)
reported the cultivation of thermophilic sludge on sucrose
for a period of 4 months. The system after adaptation was
able to take high COD loadings (86.4 kg/m3 d) and resulted
in 60% COD removal efficiency. In another study in a
thermophilic UASB reactor, Harada et al. (1996) reported
39–67% COD removal, with a corresponding BOD
removal of over 80%. The results suggested that the
wastewater contained high concentration of refractile
compounds; this, in turn, affected the microbial population
in the sludge granules. Generally, the predominant genera
of methanogens in granular sludge are Methanobacterium,
Methanobrevibacter, Methanothrix and Methanosarcina
(Bhatti et al., 1997); however, the predominance of
Methanothrix in granular sludge is most essential for the
establishment of a high performance UASB process. In this
study, abundance of Methanosarcina sp. was observed
whereas Methanothrix sp. was present to a lesser extent
thereby indicating that the latter are more sensitive to
refractile compounds (Harada et al., 1996).
4.1.1.2. Fixed bed reactor. This involves immobilization
of microorganisms on some inert support to limit the loss
of biomass and enhance the bacterial activity per unit of
reactor volume. Moreover it provides higher COD removal
at low HRT and better tolerance to toxic and organic
 ARTICLE IN PRESS

Table 4
Performance of various anaerobic reactors for molasses distillery wastewater

Reactor configuration COD loading HRT (Days) % COD % BOD References

(kg COD/ reduction reduction
m3day)

UASB 24 75 Sánchez Riera et al. (1985)

UASBa 15 2.1 90 Goodwin and Stuart (1994)
UASBb 18 490 Wolmarans and de Villiers (2002)
Thermophilic UASB Up to 86.4 60 Wiegant et al. (1985)
Thermophilic UASB Up to 28 39–67 480 Harada et al. (1996)
Thermophilic UASB Up to 30 0.3 87 Syutsubo et al. (1997)
Two-stage anaerobic treatmenta Blonskaja et al. (2003)
Anaerobic filter 2.5–5.1 10–19 54
UASB 0.6–2.5 20–39 93
Downflow fixed film reactor 14.2–20.4 3.3–2.5 85–97 60–73 Bories et al. (1988)
Two-phase thermophilic process 65 85 Yeoh (1997)
Acidogenesis 4.6–20.0 2
Methanogenesis 15.2
Diphasic (upflow) fixed film 22 3 71.8 Seth et al. (1995)
reactor (clay brick granules
support)
Diphasic (upflow) fixed film 21.3 4 67.1 Goyal et al. (1996)
reactor (granular activated
carbon support)
Upflow anaerobic filter (UAF)a 20 76 Tokuda et al. (1999)
Hybrid baffled reactor 20 77 Boopathy and Tilche (1991)
Downflow fluidized bed reactor 17 kg TOC/m3 d 0.35 75–95% TOC Garcı́a-Bernet et al. (1998)
with ground perliteb
Downflow fluidized bed reactor 4.5 3.3–1.3 85 Garcia-Calderon et al., (1998)
with ground perliteb
Downflow filter 8 55–85 Athanasopoulos (1987)
Two-stage bioreactor(anaerobic) 7 71 86 Vlissidis and Zouboulis (1993)
Ist stage (upflow sludge bed 11
reactor)
IInd stage (batch operated 0.10
bioreactor, flocculator,
precipitator)
Anaerobic contact filter (in series) 4 73–98 Vijayaraghavan and Ramanujam (2000)
Granular bed anaerobic baffled 4.75 82–90 90 Akunna and Clark (2000)
reactor (GRABBR)a
Upflow blanket filter 9–11 11–12 70 Bardiya et al. (1995)
a
Malt whisky wastewater.
b
Winery wastewater.

shock loadings. In anaerobic contact filters, various
packing materials, viz. polyurethane, clay brick, granular
activated carbon (GAC), polyvinyl chloride (PVC) plastic
media have been employed resulting in 67–98% reduction
in COD (Bories et al., 1988; Seth et al., 1995; Goyal et al.,
1996; Vijayaraghavan and Ramanujam, 2000).
GAC as support media is relatively expensive but
because of its adsorptive properties, it contributes towards
improved process stability. The interference by sulfate,
unionized sulfite and total hydrogen sulfide in anaerobic
filters is reported to be negligible (Vijayaraghavan and
Ramanujam, 2000). It was observed that the percentage
sulfate removal increased with increasing HRT from 2 to
5 d. This may be due to the utilization of sulfate as a
nutrient by microorganisms present in anaerobic contact
filter and their conversion to sulfide by sulfate reducing
bacteria (SRB) under anaerobic conditions. However at
higher sulfate concentration (426 mg/l), the removal
decreased, possibly due to low SRB population in
comparison to methanogens. Also the removal of sulfide
was explained by stripping of hydrogen sulfide from liquid
to vapor phase by the carbon dioxide and methane
generated during the anaerobic process.
In another study, Tokuda et al. (1999) performed
anaerobic treatment of undiluted whisky pot ale using an
upflow anaerobic filter (UAF) packed with special support
type (Pelia 4555, Herding GmbH, Germany) which
resulted in 76% COD removal. The pilot system also
consisted of a decanter, dephosphatation or magnesium
ammonium phosphate (MAP) (MgNH4PO4) reactor,
denitrification reactor, nitrification reactor and sedimenta-
tion tank for the reduction of nitrogen and phosphate.
Downflow filter using plastic PVC as support material
has been employed for the treatment of beet molasses
 ARTICLE IN PRESS
wastewater (Athanasopoulos, 1987). The system resulted in
55–85% reduction in COD. Also, though high sulfide
concentration (4250 mg) was inhibitory to the system, it
was not toxic at higher loadings (44 kg COD/m3 d)
probably due to high stripping of H2S.
4.1.1.3. Fluidized bed reactor. Fluidized bed reactors
contain an appropriate media such as sand, gravel or
plastics for bacterial attachment and growth. The reactors
can be operated either in the upflow or downflow modes,
with fluidization being realized by applying high fluid
velocities, normally by effluent recycling. Studies on
distillery effluent treatment in a downflow fluidized bed
system using ground perlite (an expanded volcanic rock)
resulted in 75–95% reduction in carbon content (Garcı́a-
Bernet et al., 1998). The utilization of ground perlite as a
carrier material was advantageous in the downflow
configuration because it requires low fluidization velocities
which preclude the possibility of clogging.
4.1.1.4. Two-stage processes and hybrid reactors. A two-
stage process with an anaerobic filter followed by a UASB
reactor was investigated by Blonskaja et al. (2003). The
acidogenic and methanogenic phases were clearly separated
ensuring better conditions for the methanogens. COD
reduction was 54% and 93% in the first and second stage,
respectively. In another study on a two-phase thermophilic
system, 65% COD reduction combined with a three-fold
increase in biogas yield over a single phase system was
observed (Yeoh, 1997). Boopathy and Tilche (1991) studied
the anaerobic digestion of 3–12 times diluted beet molasses
wastewater, without pH adjustment, in a hybrid anaerobic
baffled reactor (HABR). Additional nitrogen and phos-
phorus were provided in the form of urea (0.007 g/g of
COD) and diammonium hydrogen phosphate (0.0006 g/g
of COD). The reactor consisted of three chambers and a
final settler. 77% COD removal at a loading rate of
20 kg COD/m3 d was obtained.
Several variations of the UASB reactor have been
investigated for distillery wastewater treatment. In large-
scale operations, highly variable process wastewater flows
makes it difficult to maintain suitable inlet UASB flow rate;
further, prevention of the loss of low density granules is
also important. To overcome these problems, Akunna and
Clark (2000) used granular-bed anaerobic baffled reactor
(GRABBR). The reactor consisted of 10 equal compart-
ments, each of which was further divided into two with
suitable baffles. Acidogenesis was found to be predominant
in the compartments near the inlet and methanogenesis in
those located near the outlet. 82–90% COD reduction was
observed at a HRT of 4 d. Yet another modification is
upflow blanket filter (UBF) in which the packing is limited
to 5–10% height of the reactor. This configuration resulted
in 70% COD removal in sugarcane molasses distillery
spentwash (Bardiya et al., 1995).
Vlissidis and Zouboulis (1993) have investigated the
thermophilic anaerobic treatment of wastewater from the
processing of beet molasses. The process consisted of two
stages: anaerobic digestion in upflow sludge bed reactor
followed by coagulation–flocculation with lime. The HRT
was 11 d in the bioreactor and 2.5 h in the flocculator–pre-
cipitator tank. On an average, the overall treatment scheme
resulted in 86% BOD and 71% COD removal. Biogas
produced in the anaerobic reactor had a methane content
of 76%. This configuration was reported to be efficient in
treating undiluted wastewaters.
4.1.2. Aerobic treatment
The post-anaerobic treatment stage effluent still has high
organic loading and is dark brown in color, hence it is
generally followed by a secondary, aerobic treatment. Solar
drying of biomethanated spentwash is one option but the
large land area requirement limits this practice. Further, in
India, solar drying beds become non-functional during the
rainy season (Nandy et al., 2002). The other treatment
options that have been demonstrated for biomethanated
distillery effluent are described below.
 Aquaculture: Post-biomethanted effluent has been used
for pisciculture near Chennai city in southern India. The
biodigested effluent, which is a rich growth medium, is
directed to bioconversion ponds after which it is spread
in about 6 ha of fishponds. The BOD is reduced to
nearly zero and the initiative yields about 50 tons per
hectare per year of fish (Vorion Chemicals & Distilleries
Ltd., 1999).
 Constructed wetlands (CWs): Billore et al. (2001) have
demonstrated a four-celled horizontal subsurface flow
(HSF) CW for the treatment of distillery effluent after
anaerobic treatment. The post-anaerobic treated effluent
had a BOD of about 2500 mg/l and a COD of nearly
14,000 mg/l. A pre-treatment chamber filled with gravel
was used to capture the suspended solids. All the cells
were filled with gravel up to varying heights and cells
three and four supported the plants Typha latipholia and
Phragmites karka respectively. The overall retention
time was 14.4 d and the treatment resulted in 64% COD,
85% BOD, 42% total solids and 79% phosphorus
content reduction. In another study, a laboratory scale
CW employing T. latipholia was used to treat diluted
distillery effluent (Trivedy and Nakate, 2000). A root
zone of 1.5  0.3  0.3 m, filled with 75% sand and
gravel and 25% soil was used and the diluted effluent
was applied after 4 weeks of planting. The system
resulted in 76% COD reduction in 7 d which increased
marginally to 78% COD reduction in 10 d. The BOD
reduction was 22% and 47% on days 7 and 10,
respectively. In yet another instance, a distillery in
northern India is presently employing CW for polishing
the effluent prior to land discharge for irrigation in the
surrounding paddy fields (M.K. Pilania, pers. comm.).
The effluent is initially subjected to primary treatment
which includes settling and anaerobic digestion in
a structured media attached growth (SMAG)-type
 ARTICLE IN PRESS
anaerobic reactor. The primary treated effluent, with a
COD of 28,000–35,000 mg/l, is subjected to two-stage
aeration to bring down the COD to 400 mg/l. There-
after, it is directed to a CW before final discharge.
 Biocomposting: The spentwash, either directly, or after
biomethanation is sprayed in a controlled manner on
sugarcane pressmud. The latter is the filter cake
obtained during juice clarification in the manufacture
of sugar. Biocomposting is an aerobic, thermophilic
process resulting in a product rich in humus which is
thus used as a fertilizer. This is a popular option
adopted by several Indian distilleries attached to sugar
mills with adequate land availability.
The most common post-biomethanation step is the
activated sludge process wherein research efforts are
targeted at improvements in the reactor configuration
and performance. For instance, aerobic sequencing
batch reactor (SBR) is reported to be a promising solu-
tion for the treatment of effluents originating from
small wineries (Torrijos and Moletta, 1997). The treatment
system consisted of a primary settling tank, an interme-
diate retention trough, two storage tanks and an aerobic
treatment tank. A start up period of 7 d was given to
the aerobic reactor and the system resulted in 93% COD
and 97.5% BOD removal. Another configuration that
has been examined is the rotating biological reactor
(RBR) (de Bazúa et al., 1991). The system consisted of
300 l anaerobic fluidized bed reactor coupled with a 3000 l
RBR. Both the reactors were tested with diluted raw
vinasse with a COD of 60–70 g/l. At a HRT of 2 d
and COD loading up to 20 kg COD m3/day, the anaerobic
unit resulted in 70% COD removal while a lower
46% COD removal was obtained in the aerobic step.
Further testing in the aerobic system was planned with
the effluent from anaerobic reactor but no results were
reported.
The activated sludge process and its variations utilize
mixed cultures. To enhance the efficiency of aerobic
systems, several workers have focused on treatment by
pure cultures. Further, aerobic treatment has also been
examined as a precursor to anaerobic treatment. In studies
on both beet spentwash and molasses, aerobic pre-
treatment of beet spentwash with Penicillium decumbens
resulted in about 74% reduction in phenolics content and
40% reduction in color (Jiménez et al., 2003). Anaerobic
digestion without aerobic pre-treatment resulted in a sharp
drop in COD removal efficiencies with decreasing HRT.
The organic matter removal was marginally higher for beet
molasses previously fermented with P. decumbens. The
anaerobic reaction followed first-order kinetics and the rate
constant decreased on increasing the organic loading with
untreated molasses; however, it remained almost constant
with pre-treated molasses (Jiménez et al., 2003, 2004).
Geotrichum candidum is another species that resulted in
partial elimination of phenolic inhibitors such as gentisic
acid, gallic acid, quercetin, p-coumaric acid, etc., thereby
enhancing the effectiveness of anaerobic process (Borja
et al., 1993).
The following sections discuss pure culture studies on
molasses distillery wastewater targeting both COD reduc-
tion and effluent decolorization.
4.1.2.1. Fungal treatment. White rot fungus secreting
ligninolytic enzymes are capable of degrading xenobiotics
and organopollutants. Phanerochaete chrysosporium and
Trametes versicolor are the most widely studied among
these (Gonzalez et al., 2000). P. chrysosporium JAG 40
resulted in 80% decolorization of diluted synthetic
melanoidin (absorbance unit of 3.5 at 475 nm), as well as
with 6.25% anaerobically digested spentwash (Kumar et
al., 1998; Dahiya et al., 2001a). T. versicolor produces a
47 kDa extracellular enzyme identified as peroxidase which
is involved in mineralization of melanoidins. The fungus
resulted in 82% decolorization of 12.5% anaerobical-
ly–aerobically treated effluent. Of this, 90% color was
removed biologically and the rest by adsorption on the
mycelium (Dehorter and Blondeau, 1993; Benito et al.,
1997). In addition, treatment by Trametes species I-62
(CECT 20197) detoxifies the effluent by degrading furan
derivatives as observed by gas chromatography analysis
(Gonzalez et al., 2000).
Decolorization of melanoidin pigment has also been
reported by extracellular H2O2 and peroxidase produced
by Coriolus hirsutus (Miyata et al., 1998). For 6.25%
anaerobically digested spentwash, this species showed
71–75% reduction in color and 90% reduction in COD
(Kumar et al., 1998). Further, Coriolus versicolor Ps4a
decolorizes the effluent by decomposition of melanoidins
and not by the partial transformation of chromophores.
The decolorizing activity was attributed to an intracellular
enzyme which is induced in the presence of melanoidin
pigment (Aoshima et al., 1985). Treatment of biodigested
distillery wastewater by C. versicolor has also been
investigated by Chopra et al. (2004). The fungus was able
to reduce both COD and color up to 53% in 8 d; however,
glucose and peptone were required as additional nutrient
sources.
In another study, Raghukumar et al. (2004) used a
marine fungus, Flavodon flavus for the combined decolor-
ization and detoxification of 10% molasses spentwash. It
was suspected that Maillard reaction also resulted in the
formation of pyrogenic compounds like polycyclic aro-
matic hydrocarbons (PAHs) that are toxic to estuarine fish.
Treatment by F. flavus detoxified the effluent by 68%
reduction in PAH and resulted in 73% color removal. The
fungus was more effective in decolorizing raw molasses
spentwash than the anaerobically and aerobically treated
streams. This was possibly due to changes in the chemical
structure of the melanoidin pigments during anaerobic and
aerobic treatment. However, the oxygen demand of the
fungus was reportedly high.
The effects of filamentous fungi have also been studied
on distillery wastewater. These are comparatively slow
 ARTICLE IN PRESS
growing species and more susceptible to infection but the
production of a series of extracellular hydrolytic enzymes
makes it easier for them to grow on starch and cellulose
substrates. Among the filamentous species, Aspergillus sp.
is the most popular (Friedrich, 2004). A. niveus and A. niger
resulted in 60–69% reduction in color and 75–95% COD
removal; also, the treated effluent enhanced the seedling
growth in Zea mays (Angayarkanni et al., 2003; Miranda et
al., 1996). Immobilized fungal isolate of A. niger UM2
resulted in a 72% decolorization of diluted synthetic
melanoidin (absorbance unit of 3.5 at 475 nm) and 80%
decolorization of 50% biodigested effluent (Patil et al.,
2003). Similarly, a thermophilic strain of A. fumigatus G-2-
6 decolorized 75% of melanoidin pigment solution at
45 1C. Gel filtration chromatography revealed that large
molecular weight fractions of melanoidins, in particular,
were degraded rapidly (Ohmomo et al., 1987).
Apart from white-rot and filamentous fungi, yeast has
also been investigated for distillery wastewater treatment.
Yeast is characterized by quick growth and is less
susceptible to contamination by other microorganisms;
further, yeast produces biomass with high nutritive value
(Friedrich, 2004). The yeast Citeromyces WR-43-6 resulted
in high and stable removal efficiency in both color intensity
and organic matter. The removal efficiencies for diluted
spentwash (absorbance unit of 3.5 at 475 nm) were 75% for
color intensity and 76% for BOD (Sirianuntapiboon et al.,
2004a). Shojaosadati et al. (1999) optimized the growth
conditions for single cell protein (SCP) production and
COD reduction by the use of Hansenula sp. in sugar beet
stillage. They concluded that production of SCP from
stillage is one of the most promising options. Besides, the
yeast was also found to utilize lactate and acetate that are
inhibitory to ethanol production. As a result, the treated
effluent could be used as dilution water for fermentation
thereby reducing the residual stillage volume by 70%.
Another strain of Hansenula anomala J 45-N-5 and I-44
isolated from soil, resulted in 74% reduction in total
organic carbon (TOC) (Moriya et al., 1990). S. cerevisiae
also provides promising results on a larger scale. The use of
pure culture of S. cerevisiae resulted in 82.7% decoloriza-
tion in the 10% anaerobically treated distillery effluent
along with 84% reduction in COD (Selim et al., 1991;
Rajor et al., 2002). It was also reported that the nitrogen
present in the distillery effluent was sufficient for the
growth of yeast.
The dye decolorizing fungus G. candidum Dec 1
immobilized on polyurethane foam resulted in 80%
removal in color in diluted molasses solution (40–50 g/l)
(Kim and Shoda, 1999). In case of pure culture experi-
ments, Candida utilis and Trichoderma viridiae each showed
less than 65% reduction in COD whereas C. utilis and A.
niger together resulted in 89% COD removal (Nudel et al.,
1987). This reduction was from sugarcane stillage-based
media with an initial COD of 40–75 g/kg. The fungus
Mycelia sterilia D90 resulted in 91% decolorization of raw
spentwash. The color intensity (in terms of absorbance unit
at 475 nm) was originally 47 for the raw spentwash but it
was diluted to a value of 3.5 before use (Sirianuntapiboon
et al., 1988). However, a lower color removal of 60–65%
was obtained for the wastewater from anaerobic and
aerobic ponds. This was possibly due to either the
formation of some toxic compounds during anaerobic
and aerobic treatment or the inability of the strain to
attack the color causing compound due to a change in their
structure during anaerobic and aerobic treatment.
4.1.2.2. Bacterial treatment. Treatment of distillery was-
tewater by the use of Pseudomonas putida followed by
Aeromonas sp. in a two-stage bioreactor resulted in COD
as well as color reduction (Ghosh et al., 2002). P. putida
produces hydrogen peroxide which is a strong decolorizing
agent. Since the organism cannot use spentwash as a source
of carbon, 1% w/v glucose supplement was provided along
with 12.5% spentwash. Aeromonas sp. utilizes the carbo-
naceous compounds present in spentwash as the sole
carbon source, thereby eventually reducing the effluent
COD by 66% in a 24 h period. P. putida also resulted in
44% COD removal accompanied by 60% color reduction.
In another study on predigested distillery effluent with
Aeromonas formicans, 57% COD reduction and 55%
decrease in color was observed after 72 h (Jain et al.,
2000). The color removal efficiency increased up to 68%
using the bacteria immobilized on calcium alginate beads;
however, the COD reduction remained unchanged with
longer incubation period of up to 96 h.
P. fluorescence immobilized on porous cellulose carrier
resulted in 66% color removal with non-sterile diluted
spentwash (absorbance unit of 3.5 at 475 nm) and 90%
decolorization with sterile samples at 30 1C over a 4 d
period (Dahiya et al., 2001b). The decolorization efficiency
was further increased to 94% with cellulose carrier coated
with collagen. These immobilized cells could be reused but
the efficacy of color removal was reduced. In another
study, three different bacterial strains Xanthomonas fragar-
iae, Bacillus megaterium and Bacillus cereus were used both
in free form as well as after immobilization on calcium
alginate beads for the treatment of 33% predigested
distillery effluent (Jain et al., 2002). B. cereus resulted in
maximum COD (81%) and color (75%) reduction in free
form. The reduction efficiencies increased marginally with
immobilization.
The decolorization activity of acetogenic bacteria has
been reported for the first time by Sirianuntapiboon et al.
(2004b). Acetogenic bacteria is capable of oxidative
decomposition of melanoidins thereby removing low
molecular weight compounds in untreated molasses spent-
wash and almost all the low and high molecular weight
compounds in anaerobically treated molasses spentwash.
Nearly 76% decolorization, which is possibly due to a
sugar oxidase, has been observed. The nitrifying bacteria
Nitrosococcus oceanus is capable of detoxifying the
spentwash accompanied by a reduction in the chloride
content (Arora et al., 1992). However, no explanation was
 ARTICLE IN PRESS
provided for this observation. The treated wastewater leads
to better growth of rice plant due to adequate nitrogen
content and can therefore be used as a low cost fertilizer. In
yet another investigation, two aerobic bacterial strains TA
2 and TA 4 have been isolated from sites contaminated
with anaerobically treated distillery effluent (Asthana et al.,
2001). These bacteria, which were identified to be Gram-
negative and Gram-positive, respectively, resulted in 66%
and 62% BOD reduction in anaerobically treated spent-
wash. However the reduction in BOD was found to be
higher (80%) when the two were used together; further the
combination resulted in 76% color removal after 72 h.
More recently, the decolorization of four types of
synthetic melanoidins i.e., glucose–glutamic-acid (GGA),
glucose–aspartic-acid (GAA), sucrose–glutamic acid
(SGA), and sucrose–aspartic-acid (SAA), were investigated
using three different isolates, viz. Bacillus thuringiensis,
Bacillus brevis and Bacillus sp. (Kumar and Chandra,
2006). The degree of decolorization of the melanoidins
separately by each isolate was in the 1–31% range;
however, when used collectively, these isolates resulted in
up to 50% decolorization due to the enhanced effect of
coordinated metabolic interactions. The results also
indicated that the GAA polymer was the most recalcitrant
among the melanoidins tested.
The biodegradability of spentwash can be enhanced by
enzymatic pre-treatment prior to the aerobic step (Sangave
and Pandit, 2006a). After 24 h of treatment with Gram-
positive bacterium ASN6, the COD reduction of cellulase
pre-treated spentwash was 28.8% in comparison to 18.3%
for untreated effluent. This was explained by the fact that
pre-treatment affected the metabolic value (microbial
acceptability) by generating intermediate hydrolysis pro-
ducts from the parent cellulosic compounds present in the
spentwash. The biodegradability was further enhanced by
combined ultrasound and enzymatic pre-treatment result-
ing in 62.2% COD reduction after 36 h as compared to
39.4% COD removal for the untreated effluent (Sangave
and Pandit, 2006b). The enhancement in biodegradability
was attributed to molecular transformation of effluent
constituents by ultrasound pre-treatment.
4.1.2.3. Algal treatment. The treatment of anaerobically
treated 10% distillery effluent using the microalga Chlor-
ella vulgaris followed by Lemna minuscula resulted in 52%
reduction in color (Valderrama et al., 2002). In another
study, Kalavathi et al. (2001) examined the degradation of
5% melanoidin by the marine cyanobacterium Oscillatoria
boryana BDU 92181. The organism was found to release
hydrogen peroxide, hydroxyl ions and molecular oxygen
during photosynthesis resulting in 60% decolorization of
distillery effluent. In addition, this study suggested that
cyanobacteria could use melanoidin as a better nitrogen
source than carbon. Further, cyanobacteria also excrete
colloidal substances like lipopolysaccharides, proteins,
polyhydroxybutyrate (PHB), polyhydroxy-alkanoates
(PHA), etc. These compounds possess COO and ester
sulphate (OSO3 ) groups that can form complexes with
cationic sites thereby resulting in flocculation of organic
matter in the effluent. It was observed that the strain
Oscillatoria resulted in almost complete color removal
(96%) whereas Lyngbya and Synechocystis were less
effective resulting in 81 and 26% color reduction,
respectively (Patel et al., 2001). The consortium of the
three strains showed a maximum decolorization of 98%.
This was attributed to adsorption in the initial stages
followed by degradation of organic compounds which
dominated in the subsequent stages.
4.2. Physico-chemical treatment
Sugarcane molasses spentwash after biological treatment
by both anaerobic and aerobic method can still have a
BOD of 250–500 mg/l (Mall and Kumar, 1997). Also, even
though biological treatment results in significant COD
removal, the effluent still retains the dark color (Inanc et
al., 1999). The color imparting melanoidins are barely
affected by conventional biological treatment such as
methane fermentation and the activated sludge process
(Migo et al., 1993). Further, multistage biological treat-
ment reduces the organic load but intensifies the color due
to re-polymerization of colored compounds (Peña et al.,
2003). In this context, various physico-chemical treatment
options have been explored.
4.2.1. Adsorption
Activated carbon is a widely used adsorbent for the
removal of organic pollutants from wastewater but the
relatively high cost restricts its usage. Decolorization of
synthetic melanoidin using commercially available acti-
vated carbon as well as activated carbon produced from
sugarcane bagasse was investigated by Bernardo et al.
(1997). The adsorptive capacity of the different activated
carbons was found to be quite comparable. Chemically
modified bagasse using 2-diethylaminoethyl chloride hy-
drochloride and 3-chloro-2-hydroxypropyltrimethylammo-
nium chloride was capable of decolorizing diluted
spentwash (Mane et al., 2006). 0.6 g of chemically modified
bagasse in contact with 100 ml 1:4 (v/v) spentwash:water
solution resulted in 50% decolorization after 4 h contact
with intermittent swirling.
Significant decolorization was observed in packed bed
studies on anaerobically treated spentwash using commercial
activated charcoal with a surface area of 1400 m2/g (Chandra
and Pandey, 2000). Almost complete decolorization (499%)
was obtained with 70% of the eluted sample, which also
displayed over 90% BOD and COD removal. In contrast,
other workers have reported adsorption by activated carbon
to be ineffective in the treatment of distillery effluent (Sekar
and Murthy, 1998; Mandal et al., 2003). Adsorption by
commercially available powdered activated carbons resulted
in only 18% color removal; however, combined treatment
using coagulation–flocculation with polyelectrolyte followed
 ARTICLE IN PRESS
by adsorption resulted in almost complete decolorization
(Sekar and Murthy, 1998).
Low cost adsorbents such as pyorchar (activated carbon
both in granular and powdered form, manufactured from
paper mill sludge) and bagasse flyash have also been
studied for this application. Ramteke et al. (1989) reported
color removal up to 98% with pyorchar. However, to
achieve the same level of color removal, larger doses of the
indigenously prepared powdered and granular pyorchar
were required in comparison to commercial activated
carbon. Mall and Kumar (1997) compared the color
removal using commercial activated carbon and bagasse
flyash. 58% color removal was reported with 30 g/l of
bagasse flyash and 80.7% with 20 g/l of commercial
activated carbon. Since the bagasse flyash has high carbon
content and the adsorbed organic material further in-
creases its heating value, the spent adsorbent can be used
for making fire briquettes. Yet another adsorbent that has
been examined is the natural carbohydrate polymer
chitosan derived from the exoskeleton of crustaceans.
Lalov et al. (2000) studied the treatment of distillery
wastewater using chitosan as an anion exchanger. At an
optimum dosage of 10 g/l and 30 min contact time, 98%
color and 99% COD removal was observed.
4.2.2. Coagulation and flocculation
Inanc et al. (1999) reported that coagulation with alum
and iron salts was not effective for color removal. They
explored lime and ozone treatment with anaerobically
digested effluent. The optimum dosage of lime was found
to be 10 g/l resulting in 82.5% COD removal and 67.6%
reduction in color in a 30 min period. These findings are in
disagreement with those of Migo et al. (1993) who used a
commercial inorganic flocculent, a polymer of ferric
hydroxysulfate with a chemical formula [Fe2 (OH)n
(SO4)3 n/2]m for the treatment of molasses wastewater.
The treatment resulted in around 87% decolorization for
biodigested effluents; however an excess of flocculent
hindered the process due to increase in turbidity and
TOC content. FeCl3 and AlCl3 were also tested for
decolorization of biodigested effluent and showed similar
removal efficiencies. About 93% reduction in color and
76% reduction in TOC were achieved when either FeCl3 or
AlCl3 was used alone. The process was independent of
chloride and sulfate ion concentration but was adversely
affected by high fluoride concentration. However in the
presence of high flocculent concentration (40 g/l), addition
of 30 g/l CaO enhanced the decolorization process resulting
in 93% color removal. This was attributed to the ability of
calcium ions to destabilize the negatively charged mela-
noidins; further, formation of calcium fluoride (CaF2) also
precipitates the fluoride ions.
Almost complete color removal (98%) of biologically
treated distillery effluent has been reported with conven-
tional coagulants such as ferrous sulfate, ferric sulfate and
alum under alkaline conditions (Pandey et al., 2003). The
best results were obtained using Percol 47, a commercial
organic anionic polyelectrolyte, in combination with
ferrous sulfate and lime. The combination resulted in
99% reduction in color and 87 and 92% reduction in COD
and BOD, respectively. Similar findings have also been
reported by Mandal et al. (2003).
Coagulation studies on spentwash after anaerobic–aero-
bic treatment have also been conducted using bleaching
powder followed by aluminum sulfate (Chandra and Singh,
1999). The optimum dosage was 5 g/l bleaching powder
followed by 3 g/l of aluminum sulfate that resulted in 96%
removal in color, accompanied by up to 97% reduction in
BOD and COD.
Non-conventional coagulants namely wastewater from
an iron pickling industry which is rich in iron and chloride
ions and titanium ore processing industry containing
significant amounts of iron and sulfate ions have also been
examined (Pandey et al., 2003). The iron pickling waste-
water gave better results with 92% COD removal,
combined with over 98% color removal. Though the
titanium processing wastewater exhibited similar color
removal levels, the COD and BOD reductions were
perceptibly lower.
4.2.3. Oxidation process
Ozone destroys hazardous organic contaminants and has
been applied for the treatment of dyes, phenolics,
pesticides, etc. (Peña et al., 2003). Oxidation by ozone
could achieve 80% decolorization for biologically treated
spentwash with simultaneous 15–25% COD reduction. It
also resulted in improved biodegradability of the effluent.
However, ozone only transforms the chromophore groups
but does not degrade the dark colored polymeric com-
pounds in the effluent (Alfafara et al., 2000; Peña, et al.,
2003). Similarly, oxidation of the effluent with chlorine
resulted in 497% color removal but the color reappeared
after a few days (Mandal et al., 2003). Ozone in
combination with UV radiation enhanced spentwash
degradation in terms of COD; however, ozone with
hydrogen peroxide showed only marginal reduction even
on a very dilute effluent (Beltran et al., 1997).
In another study, Sangave and Pandit (2004) employed
sonication of distillery wastewater as a pre-treatment step
to convert complex molecules into a more utilizable form
by cavitation. Samples exposed to 2 h ultrasound pre-
treatment displayed 44% COD removal after 72 h of
aerobic oxidation compared to 25% COD reduction shown
by untreated samples. These results are contrary to those of
Mandal et al. (2003) who concluded ultrasonic treatment to
be ineffective for distillery spentwash treatment.
A combination of wet air oxidation and adsorption has
been successfully used to demonstrate the removal of
sulfates from distillery wastewater. Studies were done in a
counter current reactor containing 25 cm base of small
crushed stones supporting a 20 cm column of bagasse ash
as an adsorbent (Gaikwad and Naik, 2000). The waste-
water was applied from the top of the reactor and air was
supplied at the rate of 1.0 l/min. The treatment removed
 ARTICLE IN PRESS
57% COD, 72% BOD, 83% TOC and 94% sulfates. Wet
air oxidation has been recommended as part of a combined
process scheme for treating anaerobically digested spent-
wash (Dhale and Mahajani, 2000). The post-anaerobic
effluent was thermally pre-treated at 150 1C under pressure
in the absence of air. This was followed by soda-lime
treatment, after which the effluent underwent a 2 h wet
oxidation at 225 1C. 95% color removal was obtained in
this scheme. Another option is photocatalytic oxidation
that has been studied using solar radiation and TiO2 as the
photocatalyst (Kulkarni, 1998). Use of TiO2 was found to
be very effective as the destructive oxidation process leads
to complete mineralization of effluent to CO2 and H2O. Up
to 97% degradation of organic contaminants was achieved
in 90 min.
Pikaev et al. (2001) studied combined electron beam and
coagulation treatment of distillery slops from distilleries
processing grain, potato, beet and some other plant
materials. Humic compounds and lignin derivatives con-
stitute the major portion of this dark brown wastewater.
The distillery wastewater was diluted with municipal
wastewater in the ratio of 3:4, irradiated with electron
beam and then coagulated with Fe2(SO4)3. The optical
absorption in UV region was decreased by 65–70% after
this treatment. The cost was found to be less than the
existing method wherein the effluent was transported about
20 km via pipeline to a facility for biological treatment
followed by sedimentation. The treatment cost was
0.45–0.65 US$/m3 which dropped to 0.25 US$/m3 using
combined electronic-beam and coagulation method.
4.2.4. Membrane treatment
Pre-treatment of spentwash with ceramic membranes
prior to anaerobic digestion is reported to halve the COD
from 36,000 to 18,000 mg/l (Chang et al., 1994). The total
membrane area was 0.2 m2 and the system was operated at
a fluid velocity of 6.08 m/s and 0.5 bar transmembrane
pressure. In addition to COD reduction, the pre-treatment
also improved the efficiency of the anaerobic process
possibly due to the removal of inhibiting substances.
Kumaresan et al. (2003) employed emulsion liquid
membrane (ELM) technique in a batch process for
spentwash treatment. Water–oil–water type of emulsion
was used to separate and concentrate the solutes resulting
in 86% and 97% decrease in COD and BOD, respectively.
Electrodialysis has been explored for desalting spentwash
using cation and anion exchange membranes resulting in
50–60% reduction in potassium content (de Wilde, 1987).
In another study, Vlyssides et al. (1997) reported the
treatment of vinasse from beet molasses by electrodialysis
using a stainless steel cathode, titanium alloy anode and
4% w/v NaCl as electrolytic agent. Up to 88% COD
reduction at pH 9.5 was obtained; however, the COD
removal percentage decreased at higher wastewater
feeding rates.
In addition, reverse osmosis (RO) has also been
employed for distillery wastewater treatment. A unit in
western India is currently processing effluent obtained after
anaerobic digestion, followed by hold-up in a tank
maintained under aerobic conditions, in a RO system
(B.P. Agrawal, pers. comm.). 290 m3/d of RO treated
effluent is mixed with 300 m3/d of fresh water and used in
the process for various operations like molasses dilution
(290 m3/d), make-up water for cooling tower (178 m3/d),
fermenter washing (45 m3/d), etc. Yet another unit in
southern India is employing disc and tube RO modules for
direct treatment of the anaerobically digested spentwash
(M. Prabhakar Rao, pers. comm.). The permeate is
discharged while the concentrate is used for biocomposting
with sugarcane pressmud. In a recent study, Nataraj et al.
(2006) reported pilot trials on distillery spentwash using a
hybrid nanofiltration (NF) and RO process. Both the NF
and RO stages employed thin film composite (TFC)
membranes in spiral wound configuration with module
dimensions of 2.5 inches diameter and 21 inches length. NF
was primarily effective in removing the color and colloidal
particles accompanied by 80%, 95% and 45% reduction in
total dissolved solids (TDS), conductivity and chloride
concentration, respectively, at an optimum feed pressure of
30–50 bar. The subsequent RO operation at a feed pressure
of 50 bar resulted in 99% reduction each in COD,
potassium and residual TDS.
4.2.5. Evaporation/combustion
Molasses spentwash containing 4% solids can be
concentrated to a maximum of 40% solids in a quintu-
ple-effect evaporation system with thermal vapor recom-
pression (Bhandari et al., 2004; Gulati, 2004). The
condensate with a COD of 280 mg/l can be used in
fermenters. The concentrated mother liquor is spray dried
using hot air at 180 1C to obtain a desiccated powder with a
calorific value of around 3200 kcal/kg. The powder is
typically mixed with 20% agricultural waste and burnt in a
boiler. The use of recirculating fluidized bed (RCFB)
incinerator is recommended to overcome the constraints
due to stickiness of spentwash and its high sulfate content
(Alappat and Rane, 1995). Combustion is also an effective
method of on-site vinasse disposal as it is accompanied by
production of potassium-rich ash (Cortez and Peréz, 1997)
that can be used for land application.
5. Discussion
A range of biological and physico-chemical methods
have been investigated for the treatment of wastewater
from molasses-based distilleries. Because of the very high
COD, anaerobic treatment with biogas recovery is
employed extensively as the first treatment step. Anaerobic
lagoons are still used; however, most Indian distilleries
employ high rate digesters wherein the HRT is decoupled
from the SRT thereby retaining the slow growing
anaerobic microorganisms in the reactor even at high
wastewater flow. Biomethanation reduces the organic
pollution load and brings down BOD to 80–95% of the
 ARTICLE IN PRESS
original value; however, the biodigested effluent still
contains BOD in the range of 5000–10,000 mg/l (Pathade,
1999). Moreover, anaerobic treatment is a slow process and
typically requires long start-up periods. In addition, the
problem of color associated with this effluent not only
remains unsolved (Patil et al., 2003) but actually gets
aggravated since the color causing melanoidin pigment
intensifies under anaerobic conditions (Peña et al., 2003).
Therefore anaerobically treated effluent is darker in color
compared to untreated spentwash and needs several-fold
dilution by fresh water prior to discharge. The current
practice of using diluted, biodigested spentwash for
irrigation (‘‘ferti-irrigation’’) by a large number of Indian
distilleries is reportedly causing gradual soil darkening
(Kumar et al., 1998). Yet another limitation is the need to
dilute the spentwash prior to biomethanation itself. Most
distilleries follow 1–3-fold dilution to ensure proper
operation of the biomethanation plant. Since 8–15 l
spentwash is generated per liter of ethanol, dilution
increases the load on the effluent treatment plant besides
consuming considerable amount of fresh water for this
purpose.
Biological treatment using aerobic processes like acti-
vated sludge, biocomposting etc. is presently practiced by
various molasses-based distilleries. Due to the large
volumes generated, only a part of the total spentwash gets
consumed in biocomposting. Biocomposting utilizes su-
garcane pressmud as the filler material; thus it is typically
employed by distilleries attached to sugar mills. Since sugar
manufacturing is a seasonal operation, pressmud avail-
ability is often a constraint. Further, biocomposting
requires large amount of land; also, it cannot be carried
out during the rainy season.
Though aerobic treatment like the conventional acti-
vated sludge process leads to significant reduction in COD,
the process is energy intensive and the color removal is still
inadequate. Thus several pure cultures of fungi, bacteria
and algae have been investigated specifically for their
ability to decolorize the effluent as discussed earlier. In all
instances, supplementation with either nitrogen or carbon
source is almost always necessary because the microbial
species are not able to utilize the spentwash as the sole
carbon source. Further, high dilution (typically up to 1:10
fold for untreated spentwash and 1:16–1:2 fold for
biomethanated spentwash) is required for optimal micro-
bial activity. In addition, these studies are mostly limited to
laboratory scale investigations and no pilot/commercial
scale operations are reported as yet.
Physico-chemical treatment, viz. adsorption, coagula-
tion/flocculation, oxidation processes, membrane treat-
ment have been examined with particular emphasis on
effluent decolorization. Though these techniques are
effective for both color removal as well as reduction in
organic loading, sludge generation and disposal is a
constraint in coagulation/flocculation and adsorption.
Also, the cost of chemicals, adsorbents and membranes is
a deterrent to the adoption of these methods (Rajor et al.,
2002). Membrane operations like microfiltration/ultrafil-
tration for spentwash treatment are characterized by
significant membrane fouling that limits its applicability
(Jain and Balakrishnan, 2004). Decolorization through
chemical treatment with ozone and chlorine leads to
temporary color reduction because of transformation of
the chromophore groups so these are not preferred
solutions.
Thus, solutions for effective management of molasses-
based distillery wastewaters are still evolving. Some of the
gaps are highlighted below:
 Biomethanation of spentwash is well established com-
mercially; however, there is scope for several operational
improvements. These include adapting the system to
treat the spentwash without any dilution, ensuring
shorter start-up periods and degrading refractile com-
ponents to improve the anaerobic treatment efficiency.
Also, a better understanding of the re-polymerization of
color-causing compounds during anaerobic digestion
would assist in the subsequent decolorization steps.
 Biocomposting with sugarcane pressmud is increasingly
being adopted by a number of sugar complexes as a
method for disposing the biodigested spentwash. How-
ever, pressmud availability is limited; thus, alternative
materials like rice husk, wood chips, bagasse pith etc.
have been suggested (CPCB, 1994). This requires
exploring readily available local filler materials that
can be utilized for this purpose. Also, the issue of
manure quality, possibly to match the requirements of
local crops, can be addressed.
 The structure and characteristics of the color-causing
components (melanoidins) is still not fully understood.
This has consequently hindered the development of an
appropriate process scheme for their removal.
 It is established that several microorganisms (bacteria,
fungi, algae), especially in pure cultures, display a
limited ability to decolorize the spentwash. A better
understanding of the microbial enzymes/activities re-
sponsible for the degradation of melanoidins would
contribute to enhancing the efficiency of the decoloriza-
tion process.
 Investigations on pure culture aerobic systems that can
result in both COD and color removal have been
confined exclusively to laboratory scale set-ups. The
issues of appropriate system design, including scale-up,
have not been addressed. In this context, systems like
membrane bioreactors that have lower sludge produc-
tion can be considered. Also, issues like minimizing
nutrient supplementation, avoiding feed dilution and
operation under non-sterile conditions should be exam-
ined. These points are particularly significant in
translating these studies into field applications.
 Adsorbents like activated carbon that result in almost
complete decolorization are not cost effective for
treating the enormous volumes of spentwash typically
generated in a distillery. Thus, there is scope for
 ARTICLE IN PRESS
examining low cost adsorbents, including wastes gener-
ated in other industrial processes/operations. In this
context, appropriate sludge disposal methods should
also be examined.
 Membrane processes like nanofiltration and RO can
result in significant color removal thereby permitting
reuse of the treated effluent. Since spentwash is a
complex, multicomponent stream that is known to cause
considerable fouling, an understanding of the compo-
nents those are primarily responsible for this phenom-
enon would assist in appropriate feed pre-treatment, for
the efficient operation of the membrane system.
6. Conclusion
The review indicates that a comprehensive treatment
scheme for molasses distillery wastewater leading to
effective removal of both organics and color is not
currently available. The properties of the color-causing
melanoidins and their transformation during anaerobic
treatment are also not fully understood. Biological treat-
ment, especially with pure cultures, appears promising and
possibly cost-effective for color removal; however, the
initiatives are mainly confined to laboratory trials. Physico-
chemical methods are capable of both color and organic
load reduction; consequently, in spite of the cost, even
high-end options like membrane filtration are being field-
tested. There is thus an urgent need to address the
limitations in the existing methods and to develop
integrated treatment processes that provide a complete
solution to the treatment of wastewater from molasses-
based distilleries.
Acknowledgement
Y. Satyawali gratefully acknowledges the financial
support in the form of Junior Research Fellowship
provided by University Grants Commission, New Delhi,
India.
References
Akunna, J.C., Clark, M., 2000. Performance of a granular-bed anaerobic
baffled reactor (GRABBR) treating whisky distillery wastewater.
Bioresource Technology 74 (3), 257–261.
Alappat, B.J., Rane, V.C., 1995. An algorithm to calculate the
performance details of an RCFB incinerator with heat recovery for
the treatment of distillery spentwash. International Journal of Energy
Research 19, 329–336.
Alfafara, C.G., Migo, V.P., Amrante, J.A., Dallo, R.F., Matsumara, M.,
2000. Ozone treatment of distillery slop waste. Water Science and
Technology 42 (3–4), 193–198.
Angayarkanni, J., Palaniswami, M., Swaminathan, K., 2003. Biotreat-
ment of distillery effluent using Aspergillus niveus. Bulletin of
Environmental Contamination and Toxicology 70 (2), 268–277.
Aoshima, I., Tozawa, Y., Ohmomo, S., Ueda, K., 1985. Production of
decolorizing activity for molasses pigment by Coriolus versicolor Ps4a.
Agricultural and Biological Chemistry 49 (7), 2041–2045.
Arora, M., Sharma, D.K., Behera, B.K., 1992. Upgrading of distillery
effluent by Nitrosococcus oceanus for its use as a low-cost fertilizer.
Resources, Conservation and Recycling 6 (4), 347–353.
Asthana, A.K., Misra, S.K., Chandra, R., Guru, R., 2001. Treatment of
color and biochemical oxygen demand of anaerobically treated
distillery effluent by aerobic bacterial strains. Indian Journal of
Environmental Protection 21 (12), 1070–1072.
Athanasopoulos, N., 1987. Anaerobic treatment of beet molasses
alcoholic fermentation wastewater in a downflow filter. Resources
and Conservation 15 (1–2), 147–150.
Bardiya, M.C., Hashia, R., Chandna, S., 1995. Performance of hybrid
reactor for anaerobic digestion of distillery effluent. Journal of Indian
Association for Environment Management 22 (3), 237–239.
Baruah, A.K., Sharma, R.N., Borah, G.C., 1993. Impact of sugar mill and
distillery effluents on water quality of river Gelabil, Assam. Indian
Journal of Environmental Health 35 (4), 288–293.
Beltran, F.J., Encinar, J.M., González, J.F., 1997. Industrial waste-
water advanced oxidation. Part 2. Ozone combination with
hydrogen peroxide or UV radiation. Water Research 31 (10),
2415–2428.
Benito, G.G., Miranda, M.P., de los Santos, D.R., 1997. Decolorization of
wastewater from an alcoholic fermentation process with Trametes
versicolor. Bioresource Technology 61 (1), 33–37.
Berg, C., 2004. World fuel ethanol analysis and outlook /http://
www.distill.com/World-Fuel-Ethanol-A&O-2004.htmlS (accessed
10.07.2006).
Bernardo, E.C., Egashira, R., Kawasaki, J., 1997. Decolorization of
molasses wastewater using activated carbon prepared from cane
bagasse. Carbon 35 (9), 1217–1221.
Bhandari, H.C., Mitra, A.K., Kumar, S., 2004. Crest’s integrated system:
reduction and recycling of effluents in distilleries. In: Tewari, P.K.
(Ed.), Liquid Asset, Proceedings of Indo-EU Workshop on Promoting
Efficient Water Use in Agro-based Industries. TERI Press, New Delhi,
India, pp. 167–169.
Bhatti, Z.I., Furukawa, K., Fujita, M., 1997. Microbial diversity in UASB
reactors. Pure and Applied Chemistry 69 (11), 2431–2438.
Billore, S.K., Singh, N., Ram, H.K., Sharma, J.K., Singh, V.P., Nelson,
R.M., Dass, P., 2001. Treatment of molasses based distillery effluent in
a constructed wetland in central India. Water Science and Technology
44 (11–12), 441–448.
Blonskaja, V., Menert, A., Vilu, R., 2003. Use of two-stage anaerobic
treatment for distillery waste. Advances in Environmental Research 7
(3), 671–678.
Boopathy, R., Tilche, A., 1991. Anaerobic digestion of high strength
molasses wastewater using hybrid anaerobic baffled reactor. Water
Research 25 (7), 785–790.
Bories, A., Raynal, J., Bazile, F., 1988. Anaerobic digestion of high-
strength distillery wastewater (cane molasses stillage) in a fixed-film
reactor. Biological Wastes 23 (4), 251–267.
Borja, R., Martı́n, A., Maestro, R., Luque, M., Durán, M.M., 1993.
Enhancement of the anaerobic digestion of wine distillery wastewater
by the removal of phenolic inhibitors. Bioresource Technology 45 (2),
99–104.
Central Pollution Control Board (CPCB), 1994./http://www.cpcb.nic.in/
dec942.htmS (accessed on 21.07.2006).
Central Pollution Control Board (CPCB), 2003. Environmental Manage-
ment in Selected Industrial Sectors Status and Needs, PROBES/97/
2002–03, CPCB, Ministry of Environment and Forest, New Delhi.
Chandra, R., Pandey, P.K., 2000. Decolorization of anaerobically treated
distillery effluent by activated charcoal adsorption method. Indian
Journal of Environmental Protection 21 (2), 134–137.
Chandra, R., Singh, H., 1999. Chemical decolorization of anaerobically
treated distillery effluent. Indian Journal of Environmental Protection
19 (11), 833–837.
Chang, I.S., Choo, K.H., Lee, C.H., Pek, U.H., Koh, U.C., Kim, S.W.,
Koh, J.H., 1994. Application of ceramic membrane as a pre-treatment
in anaerobic digestion of alcohol-distillery wastes. Journal of
Membrane Science 90 (1–2), 131–139.
 ARTICLE IN PRESS
Chen, J.C.P., Chou, C.C., 1993. Cane Sugar Handbook. Wiley, New
York, pp. 408–409.
Chopra, P., Singh, D., Verma, V., Puniya, A.K., 2004. Bioremediation of
melanoidin containing digested spentwash from cane-molasses dis-
tillery with white rot fungus Coriolus versicolor. Indian Journal of
Microbiology 44 (3), 197–200.
Cortez, L.A.B., Peréz, L.E.B., 1997. Experiences on vinasse disposal: Part
III: combustion of vinasse-]6 fuel oil emulsions. Brazilian Journal of
Chemical Engineering [online] 14 (1) [cited 24 January 2006] Available
from World Wide Web: /http://www.scielo.br/scielo.php?script=
sci_arttext&pid=S0104-66321997000100002&lng=en&nrm=isoS.
ISSN 0104-6632. (accessed 21.07.2006).
Curtin, L.V., 1983. Molasses—general considerations. In: Molasses in
Animal Nutrition. National Feed Ingredients Association, West Des
Moines, IA, pp. 1–11.
Dahiya, J., Singh, D., Nigam, P., 2001a. Decolorization of synthetic and
spentwash melanoidins using the white-rot fungus Phanerochaete
chrysosporium JAG-40. Bioresource Technology 78 (1), 95–98.
Dahiya, J., Singh, D., Nigam, P., 2001b. Decolorization of molasses
wastewater by cells of Pseudomonas fluorescens immobilized on porous
cellulose carrier. Bioresource Technology 78 (1), 111–114.
de Bazúa, C.D., Cabrero, M.A., Poggi, H.M., 1991. Vinasses biological
treatment by anaerobic and aerobic processes: laboratory and pilot-
plant tests. Bioresource Technology 35 (1), 87–93.
de Wilde, F.G.N., 1987. Demineralization of a molasses distillery
wastewater. Desalination 67, 481–493.
Dehorter, B., Blondeau, R., 1993. Isolation of extracellular Mn-dependent
enzyme mineralizing melanoidins from the white rot fungus Trametes
versicolor. FEMS Microbiology Letters 109, 117–122.
Dhale, A.D., Mahajani, V.V., 2000. Treatment of distillery waste after
bio-gas generation: wet oxidation. Indian Journal of Chemical
Technology 7, 11–18.
Dhar, G.M., Thampli, J., Pandit, A.B., Lele, S.S., Joshi, J.B., 1998a.
Overall treatment of thermally pre-treated distillery waste Part I.
Indian Chemical Engineer, Section A 40 (3), 222–231.
Dhar, G.M., Thampli, J., Pandit, A.B., Lele, S.S., Joshi, J.B., 1998b.
Overall treatment of thermally pre-treated distillery waste Part II.
Indian Chemical Engineer, Section A 40 (3), 232–240.
FitzGibbon, F., Singh, D., McMullan, G., Marchant, R., 1998. The effect
of phenolic acids and molasses spentwash concentration on distillery
wastewater remediation by fungi. Process Biochemistry 33 (8),
799–803.
Friedrich, J., 2004. Bioconversion of distillery waste. In: Arora, D.K.
(Ed.), Fungal Biotechnology in Agricultural, Food and Environmental
Applications. Marcel Dekker Inc., New York, pp. 431–442.
Gaikwad, R.W., Naik, P.K., 2000. Technology for the removal of sulfate
from distillery wastewater. Indian Journal of Environmental Protec-
tion 20 (2), 106–108.
Garcı́a-Bernet, D., Buffiére, P., Elmaleh, S., Moletta, R., 1998. Applica-
tion of the down-flow fluidized bed to the anaerobic treatment of wine
distillery wastewater. Water Science and Technology 38 (8–9),
393–399.
Garcia-Calderon, D., Buffiére, P., Moletta, R., Elmaleh, S., 1998.
Anaerobic digestion of wine distillery wastewater in down-flow
fluidized bed. Water Research 32 (12), 3593–3600.
Ghosh, M., Ganguli, A., Tripathi, A.K., 2002. Treatment of anaerobically
digested distillery spentwash in a two-stage bioreactor using Pseudo-
monas putida and Aeromonas sp. Process Biochemistry 37 (8), 857–862.
Godbole, J., 2002. Ethanol from cane molasses, Fuel Ethanol Workshop,
Honululu, Hawaii, November 14, 2002. /http://www.hawaii.gov/
dbedt/ert/new-fuel/files/ethanol-workshop/10-Godbole-DOE-HI.pdfS
(accessed on 9.08.2006)
Godshall, M.A., 1999. Removal of colorants and polysaccharides and the
quality of white sugar. In: Proceedings of sixth International
Symposium Organized by Association Andrew van Hook (AvH),
March 25 1999, Reims, France, pp. 28–35.
Gonzalez, T., Terron, M.C., Yague, S., Zapico, E., Galletti, G.C.,
Gonzalez, A.E., 2000. Pyrolysis/gas chromatography/ mass spectro-
metry monitoring of fungal-biotreated distillery wastewater using
Trametes sp. I-62 (CECT 20197). Rapid Communications in Mass
Spectrometry 14 (15), 1417–1424.
Goodwin, J.A.S., Stuart, J.B., 1994. Anaerobic digestion of malt whisky
distillery pot ale using upflow anaerobic sludge blanket reactors.
Bioresource Technology 49 (1), 75–81.
Goyal, S.K., Seth, R., Handa, B.K., 1996. Diphasic fixed-film biometha-
nation of distillery spentwash. Bioresource Technology 56 (2–3),
239–244.
Gulati, N., 2004. Conservation of resources using evaporation and spray
drying technology for distillery and paper industries. In: Tewari, P.K.
(Ed.), Liquid Asset, Proceedings of Indo-EU Workshop on Promoting
Efficient Water Use in Agro-Based Industries. TERI Press, New Delhi,
India, pp. 163–166.
Gunasekaran, P., Raj, K.C., 1999. Ethanol fermentation technology—
Zymomonas mobilis. Current Science India 77 (1), 56–68.
Harada, H., Uemura, S., Chen, A.C., Jayadevan, J., 1996. Anaerobic
treatment of a recalcitrant wastewater by a thermophilic UASB
reactor. Bioresource Technology 55 (3), 215–221.
Inanc, B., Ciner, F., Ozturk, I., 1999. Color removal from fermen-
tation industry effluents. Water Science and Technology 40 (1),
331–338.
Jain, S., Balakrishnan, M., 2004. Membrane fouling and cleaning in
ultrafiltration of anaerobically treated distillery effluent. In: Euro-
membrane 2004, Hamburg, Germany, 28 September–1 October 2004.
Jain, N., Prajapati, S.K., Minocha, A.K., Verma, C.L., 2000. Batch
studies on the degradation of spentwash from distilleries. Indian
Journal of Environmental Protection 21 (2), 122–126.
Jain, N., Minocha, A.K., Verma, C.L., 2002. Degradation of predigested
distillery effluent by isolated bacterial strains. Indian Journal of
Experimental Biology 40, 101–105.
Jiménez, A.M., Borja, R., Martı́n, A., 2003. Aerobic–anaerobic biode-
gradation of beet molasses alcoholic fermentation wastewater. Process
Biochemistry 38 (9), 1275–1284.
Jiménez, A.M., Borja, R., Martı́n, A., 2004. A comparative kinetic
evaluation of the anaerobic digestion of untreated molasses and
molasses previously fermented with Penicillium decumbens in batch
reactors. Biochemical Engineering Journal 18 (2), 121–132.
Joshi, H.C., 1999. Bio-energy potential of distillery effluents. Bio Energy
News 3 (3), 10–15.
Kalavathi, D.F., Uma, L., Subramanian, G., 2001. Degradation and
metabolization of the pigment- melanoidin in a distillery effluent by
the marine cyanobacterium Oscillatoria boryana BDU 92181. Enzyme
and Microbial Technology 29 (4–5), 246–251.
Kim, S.J., Shoda, M., 1999. Batch decolorization of molasses by
suspended and immobilized fungus of Geotrichum Candidum Dec 1.
Journal of Bioscience and Bioengineering 88 (5), 586–589.
Kulkarni, A.K., 1998. Solar assisted photocatalytic oxidation of distillery
waste. Indian Chemical Engineer 40 (2), 169–172.
Kumar, P., Chandra, R., 2006. Decolourisation and detoxification of
synthetic molasses melanoidins by individual and mixed cultures of
Bacillus spp. Bioresource Technology 97 (16), 2096–2102.
Kumar, V., Wati, L., Nigam, P., Banat, I.M., Yadav, B.S., Singh, D.,
Marchant, R., 1998. Decolorization and biodegradation of anaerobi-
cally digested sugarcane molasses spentwash effluent from bio-
methanation plants by white-rot fungi. Process Biochemistry 33 (1),
83–88.
Kumaresan, T., Sheriffa Begum, K.M.M., Sivashanmugam, P.,
Anantharaman, N., Sundaram, S., 2003. Experimental studies on
treatment of distillery effluent by liquid membrane extraction.
Chemical Engineering Journal 95 (1–3), 199–204.
Lalov, I.G., Guerginov, I.I., Krysteva, A., Fartsov, K., 2000. Treatment of
wastewater from distilleries with chitosan. Water Research 34 (5),
1503–1506.
Lata, K., Kansal, A., Balakrishnan, M., Rajeshwari, K.V., Kishore,
V.V.N., 2002. Assessment of biomethanation potential of selected
industrial organic effluents in India. Resources, Conservation and
Recycling 35 (3), 147–161.
 ARTICLE IN PRESS
Lin, Y., Tanaka, S., 2006. Ethanol fermentation from biomass resources:
current state and prospects. Applied Microbiology Biotechnology 69,
627–642.
Mahimairaja, S., Bolan, N.S., 2004. Problems and prospects of
agricultural use of distillery spentwash in India. In: Third Australian
and New Zealand Soil Science Societies Joint Conference, Sydney,
Australia, 5–9 December 2004.
Mall, I.D., Kumar, V., 1997. Removal of organic matter from distillery
effluent using low cost adsorbent. Chemical Engineering World XXXII
(7), 89–96.
Mandal, A., Ojha, K., Ghosh, D.N., 2003. Removal of color from
distillery wastewater by different processes. Indian Chemical Engineer
Section B 45 (4), 264–267.
Mane, J.D., Modi, S., Nagawade, S., Phadnis, S.P., Bhandari, V.M., 2006.
Treatment of spentwash using chemically modified bagasse and color
removal studies. Bioresource Technology 97 (14), 1752–1755.
Martins, S.I.F.S., van Boekel, M.A.J.S., 2004. A kinetic model for the
glucose/glycine Maillard reaction pathways. Food Chemistry 90 (1–2),
257–269.
Migo, V.P., Matsumara, M., Rosario, E.J.D., Kataoka, H., 1993.
Decolorization of molasses wastewater using an inorganic flocculant.
Journal of Fermentation and Bioengineering 75 (6), 438–442.
Miranda, M.P., Benito, G.G., Cristobal, N.S., Nieto, C.H., 1996. Color
elimination from molasses wastewater by Aspergillus niger. Bior-
esource Technology 57 (3), 229–235.
Mitsui & Co., 2003. Nano-Research Institute, XNRI (Inc.). /http://
www.xnri.com/English/rd/pdf/ZEOLITE.pdfS (accessed on
21.07.2006).
Miyata, N., Iwahori, K., Fujita, M., 1998. Manganese-independent and
manganese-dependent decolorization of melanoidin by extracellular
hydrogen peroxide and peroxidases from Coriolus hirsutus pellets.
Journal of Fermentation and Bioengineering 85 (5), 550–553.
Moriya, K., Iefuji, H., Shimoi, H., Sato, S.I., Tadenuma, M., 1990.
Treatment of distillery wastewater discharged from beet molasses-
spirits production using yeast. Journal of Fermentation and Bioengi-
neering 69 (2), 138–140.
Nandy, T., Shastry, S., Kaul, S.N., 2002. Wastewater management in cane
molasses distillery involving bioresource recovery. Journal of Envir-
onmental Management 65 (1), 25–38.
Nataraj, S.K., Hosamani, K.M., Aminabhavi, T.M., 2006.
Distillery wastewater treatment by the membrane-based nano-
filtration and reverse osmosis processes. Water Research 40 (12),
2349–2356.
Nudel, B.C., Waehner, R.S., Fraile, E.R., Giulietti, A.M., 1987. The use of
single and mixed cultures for aerobic treatment of cane sugar stillage
and SCP production. Biological Wastes 22 (1), 67–73.
Ohmomo, S., Kaneko, Y., Sirianuntapiboon, S., Atthasampunna, P.,
Nakamura, I., 1987. Decolorization of molasses wastewater by
thermophilic strain Aspergillus fumigatus G-2-6. Agricultural and
Biological Chemistry 51 (12), 3339–3346.
Pandey, R.A., Malhotra, S., Tankhiwale, A., Pande, S., Pathe, P.P.,
Kaul, S.N., 2003. Treatment of biologically treated distillery effluent—
a case study. International Journal of Environmental Studies 60 (3),
263–275.
Patel, A., Pawar, R., Mishra, S., Tewari, A., 2001. Exploitation of marine
cyanobacteria for removal of color from distillery effluent. Indian
Journal of Environmental Protection 21 (12), 1118–1121.
Pathade, G.R., 1999. A review of current technologies for distillery
wastewater treatment. In: Goel, P.K. (Ed.), Advances in Industrial
Wastewater Treatment. Technoscience Publications, Rajasthan, India,
pp. 180–239.
Patil, P.U., Kapadnis, B.P., Dhamankar, V.S., 2003. Decolorization of
synthetic melanoidin and biogas effluent by immobilized fungal isolate
of Aspergillus niger UM2, All India Distiller’s Association (AIDA)
Newsletter 53–56.
Peña, M., Coca, M., González, R., Rioja, R., Garcı́a, M.T., 2003.
Chemical oxidation of wastewater from molasses fermentation with
ozone. Chemosphere 51 (9), 893–900.
Picataggio, S.K., Zhang, M., Eddy, C.K., Deanda, K.A., 1996.
Recombinant Zymomonas for pentose fermentation. United States
Patent 5726053.
Pikaev, A.K., Ponomarev, A.V., Bludenko, A.V., Minin, V.N., Elizar’eva,
L.M., 2001. Combined electronic-beam and coagulation purification of
molasses distillery slops. Features of the method, technical and
economic evaluation of large scale facility. Radiation Physics and
Chemistry 61 (1), 81–87.
Raghukumar, C., Mohandass, C., Kamat, S., Shailaja, M.S., 2004.
Simultaneous detoxification and decolorization of molasses spentwash
by the immobilized whit-rot fungus Flavodon flavus isolated from the
marine habitat. Enzyme and Microbial Technology 35 (2–3), 197–202.
Rajor, R., Singh, R., Mathur, R.P., 2002. Color removal of distillery waste
by Sacchromyces. Indian Journal of Environmental Protection 22 (12),
1241–1252.
Ramteke, D.S., Wate, S.R., Moghe, C.A., 1989. Comparative adsorption
studies of distillery waste on activated carbon. Indian Journal of
Environmental Health 31 (1), 17–24.
Rivero-Pérez, M.D., Pérez-Magariño, S., José, M.L., 2002. Role of
melanoidins in sweet wines. Analytica Chimica Acta 458 (1), 169–175.
Rodrı́guez, J.G.O., 2000. Effects of vinasse on sugarcane (saccharum
officinarum) productivity. Revista de la Facultad de Agronomia,
Universidad del Zulia 17, pp. 318–326 /http://www.redpav-fpolar.-
info.ve/fagroluz/Archivo%20de%20Acrobac/17-4/ra4006%20ab-
stract.pdfS (accessed on 9.08.2006).
Sánchez Riera, F., Córdoba, P., Siñeriz, F., 1985. Use of the UASB
reactor for the anaerobic treatment of stillage from sugarcane
molasses. Biotechnology and Bioengineering 27 (12), 1710–1716.
Sangave, P.C., Pandit, A.B., 2004. Ultrasound pre-treatment for enhanced
biodegradability of the distillery wastewater. Ultrasonics Sonochem-
istry 11 (3–4), 197–203.
Sangave, P.C., Pandit, A.B., 2006a. Enhancement in biodegradability of
distillery wastewater using enzymatic pre-treatment. Journal of
Environmental Management 78, 77–85.
Sangave, P.C., Pandit, A.B., 2006b. Ultrasound and enzyme assisted
biodegradation of distillery wastewater. Journal of Environmental
Management 80 (1), 36–46.
Sekar, D., Murthy, D.V.S., 1998. Color removal of distillery spentwash by
adsorption technique. Indian Chemical Engineer, Section A 40 (4),
176–181.
Selim, M.H., Elshafei, A.M., El-Diwany, A.I., 1991. Production of single
cell protein from yeast strains grown in Egyptian vinasse. Bioresource
Technology 36 (2), 157–160.
Sennitt, T., 2005. Emissions and economics of biogas and power. In: 68th
Annual Water Industry Engineers and Operators’ Conference,
Schweppes Centre, Bendigo, 7 and 8 September 2005.
Seth, R., Goyal, S.K., Handa, B.K., 1995. Fixed film biomethanation of
distillery spentwash using low cost porous media. Resources,
Conservation and Recycling 14 (2), 79–89.
Sheehan, G.J., Greenfield, P.F., 1980. Utilization, treatment and disposal
of distillery wastewater. Water Research 14 (3), 257–277.
Shojaosadati, S.A., Khalilzadeh, R., Jalilzadeh, A., Sanaei, H.R., 1999.
Bioconversion of molasses stillage to protein as an economic treatment
of this effluent. Resources, Conservation and Recycling 27 (1–2),
125–138.
Sirianuntapiboon, S., Somchal, P., Sihanonth, P., Atthasampunna, P.,
Ohmomo, S., 1988. Microbial decolorization of molasses wastewater
by Mycelia sterilia D90. Agricultural and Biological Chemistry 52 (2),
393–398.
Sirianuntapiboon, S., Zohsalam, P., Ohmomo, S., 2004a. Decolorization
of molasses wastewater by Citeromyces sp. WR-43-6. Process
Biochemistry 39 (8), 917–924.
Sirianuntapiboon, S., Phothilangka, P., Ohmomo, S., 2004b. Decoloriza-
tion of molasses wastewater by strain no. BP103 of acetogenic bacteria.
Bioresource Technology 92 (1), 31–39.
Siva Kesava, S., Panda, T., Rakshit, S.K., 1996. Production of ethanol by
immobilized whole cells of Zymomonas mobilis in an expanded bed
bioreactor. Process Biochemistry 31 (5), 449–456.
 ARTICLE IN PRESS
Skerratt, G., 2004. European distilleries: an overview. In: Tewari, P.K.
(Ed.), Liquid Asset, Proceedings of the Indo-EU Workshop on
Promoting Efficient Water Use in Agro-Based Industries. TERI Press,
New Delhi, India, pp. 1–11.
Subba Rao, B., 1972. A low cost waste treatment method for the disposal
of distillery waste (spentwash). Water Research 6 (11), 1275–1282.
Subramanian, K.A., Singal, S.K., Saxena, M., Singhal, S., 2005.
Utilization of liquid biofuels in automotive diesel engines: an Indian
perspective. Biomass and Bioenergy 29 (1), 65–72.
Syutsubo, K., Harada, H., Ohashi, A., Suzuki, H., 1997. An effective
start-up of thermophilic UASB reactor by seeding mesophilically-
grown granular sludge. Water Science and Technology 36 (6–7),
391–398.
Tano, M.S., Buzato, J.B., 2003. Effect of the presence of initial ethanol on
ethanol production in sugarcane juice fermented by Zymomonas
mobilis. Brazilian Journal of Microbiology 34, 242–244.
Tao, F., Miao, J.Y., Shi, G.Y., Zhang, K.C., 2005. Ethanol fermentation
by an acid-tolerant Zymomonas mobilis under non-sterilized condition.
Process Biochemistry 40 (1), 183–187.
Tokuda, M., Fujiwara, Y., Kida, K., 1999. Pilot plant test for the removal
of organic matter, N and P from whisky pot ale. Process Biochemistry
35 (3–4), 267–275.
Toma, M.M., Kalnenieks, U., Berzins, A., Vigants, A., Rikmanis, M.,
Viesturs, U., 2003. The effect of mixing on glucose fermentation by
Zymomonas mobilis continuous culture. Process Biochemistry 38 (9),
1347–1350.
Torrijos, M., Moletta, R., 1997. Winery wastewater depollution by
sequencing batch reactor. Water Science and Technology 35 (1),
249–257.
Trivedy, R.K., Nakate, S.S., 2000. Treatment of diluted distillery waste by
constructed wetlands. Indian Journal of Environmental Protection 20
(10), 749–753.
Uppal, J., 2004. Water utilization and effluent treatment in the Indian
alcohol industry: an overview. In: Tewari, P.K. (Ed.), Liquid Asset,
Proceedings of the Indo-EU Workshop on Promoting Efficient Water
Use in Agro-Based Industries. TERI Press, New Delhi, India, pp.
13–19.
Valderrama, L.T., Del Campo, C.M., Rodriguez, C.M., de- Bashan, L.E.,
Bashan, Y., 2002. Treatment of recalcitrant wastewater from ethanol
and citric acid production using the microalga Chlorella vulgaris
and the macrophyte Lemna minuscula. Water Research 36 (17),
4185–4192.
Vijayaraghavan, K., Ramanujam, T.K., 2000. Performance of anaerobic
contact filter in series for treating distillery spentwash. Bioprocess and
Biosystems Engineering 22 (2), 109–114.
Vlissidis, A., Zouboulis, A.I., 1993. Thermophilic anaerobic digestion of
alcohol distillery wastewaters. Bioresource Technology 43 (2),
131–140.
Vlyssides, A.G., Israilides, C.J., Loizidou, M., Karvouni, G., Mourafeti,
V., 1997. Electrochemical treatment of vinasse from beet molasses.
Water Science and Technology 36 (2–3), 271–278.
Vorion Chemicals & Distilleries Ltd., 1999. Distillery Effluent Treatment
and Gold Fish Culture Project, /http://www.ias.unu.edu/proceedings/
icibs/ibs/info/india/vorion/index.htmS (accessed 9.08.2006).
Wiegant, W.M., Claassen, J.A., Lettinga, G., 1985. Thermophilic
anaerobic digestion of high strength wastewaters. Biotechnology and
Bioengineering 27, 1374–1381.
Wilkie, A.C., Riedesel, K.J., Owens, J.M., 2000. Stillage characterization
and anaerobic treatment of ethanol stillage from conventional and
cellulosic feedstocks. Biomass and Bioenergy 19 (2), 63–102.
Wolmarans, B., de Villiers, G.H., 2002. Start-up of a UASB treatment
plant on distillery wastewater. Water SA 28 (1), 63–68 /http://www.
wrc.org.za/publications/watersa/2002/januaury/1393.pdfS (accessed
9.08.2006).
Yeoh, B.G., 1997. Two-phase anaerobic treatment of cane-molasses
alcohol stillage. Water Science and Technology 36 (6–7), 441–448.
Zhang, M., 2000. Recombinant Zymomonas mobilis with improved xylose
utilization, United States Patent 6566107.