Recent Research in Science and Technology 2011, 3(4): 169-176

ISSN: 2076-5061
www.recent-science.com
BOTANY

EFFECT OF DISTILLERY SPENT WASH (DSW) AND FERTILIZER ON

GROWTH AND CHLOROPHYLL CONTENT OF SUGARCANE (SACCHARUM
OFFICINARUM L.) PLANT
P.Rath1, G.Pradhan1, M.K.Misra2
1Department
2PG

of Botany, Aska Science College, Berhampur University, Aska, Ganjam 761111, Odisha
Department of Botany, Berhampur University, Berhampur, 760007, Odisha

Abstract
A field experiment was conducted with distillery spent wash (DSW) collected from Aska Co-operative Sugar Industries Ltd., Aska, Odisha,
India and other two types of treatments viz; inorganic fertilizer and cow dung to observe a comparative effect on growth and chlorophyll
content using sugarcane variety Co1274 as the test crop. The experiment was formulated with four treatments (50% DSW, 100% DSW,
fertilizer & fertilizer + cow dung) each with three replicates and one set of control. The growth parameters such as height of the plant, length
and girth of stem, breadth of leaves, number of leaves and number of tillers per plant, leaf area index and total chlorophyll content of
sugarcane plant were estimated during different growth periods in all treatments. All the parameters showed an increasing trend from the
control except 100% distillery spent wash, where a declining trend over the control in all parameters was observed. Maximum growth and
chlorophyll content was observed in 50% distillery spent wash as compared with two different types of fertilizers.
Keywords: Distillery spent wash (DSW), Sugarcane growth, Height, Stem girth, Chlorophyll content
Introduction
One of the most important environmental problems faced
by the world is management of wastes. Different industries
create a variety of waste water pollutants; which are difficult and
costly to treat. Waste water characteristics and levels of
pollutants vary significantly from industry to industry. The use of
industrial waste as soil amendment has generated interest in
recent time. The waste water produced continuously could cater
the needs of irrigated crops [1].
Production of ethyl alcohol in distilleries based on
sugarcane molasses constitutes a major industry in Asia and
South America. The worlds total annual production of alcohol
from sugarcane molasses is more than 13 million m3. The
aqueous distillery effluent stream known as spent wash is a dark
brown highly organic effluent and is approximately 12-15 times
by volume of product alcohol [2]. The disposal of distillery spent
wash is of serious concern due to its large volume and high
biological oxygen demand (BOD) and chemical oxygen demand
(COD). Due to high concentration of organic load, distillery spent
wash is a potential source of renewable energy. The effluent
does not contain any toxic heavy metals as it is a waste from
plant materials. It contains high amount of nutrients such as
nitrogen, phosphorous, potassium, sulphur and a large amount
of micronutrients. The land application of distillery spent wash
often benefits water pollution control and utilization for
agricultural production [3]. So it can be applied directly to the
land as irrigation water as it helps in restoring and maintaining
soil fertility, increasing soil microflora, improving physical and
chemical properties of soil leading to better water retaining
capacity of the soil. The effluent is ideal for sugarcane, maize,
wheat and rape seed production [4]. It has been reported that
waste water from different industries produced continuously
could cater the needs of irrigated crops [5]. Thus the distillery
spent wash will not only prevent waste from being an
environmental hazard but also served as an additional potential
source of fertilizer for agricultural use. Diluted spent wash
increased the growth of shoot length, leaf number per plant, leaf
area and chlorophyll content of peas [6]. It was also reported

that the water holding capacity, cation exchange capacity,

increases the availability of nitrogen, phosphorus, potassium,
copper, zinc, iron, manganese; but with reduced biochemical
oxygen demand (BOD) with addition of sewage sludge to a
course textured sandy and calcareous soil [7]. An increase in
the soil organic matter by 1% with sugar factory effluent applied
to soils was observed in Cuba [8]. Many workers reported an
adverse effect of higher concentration of different types of
industrial effluents in the growth rate of different crops [9, 10, 11,
12].
There have been studies related to the application of
distillery spent wash to agriculture in India as well as other parts
of the world. Spent wash at the rate of 35-50 m3 ha-1 was
recommended as optimum dose for higher sugarcane yield in
Brazil and in Australia. In Sau Paulo, Brazil, the crop productivity
was 2-10 times higher as compared to the untreated lands.
Distillery spent wash was found to increase the cane yield in
sugarcane and decrease the potassium fertilizer need in a study
conducted by Caroni Research Station, Trinidad and Tobago. In
Philippines, spent wash application at the rate of 80-240 m3 ha-1
in addition to chemical fertilizers increased the cane yield by 1012 percent and sugar yield by 13-16 percent compared to
normal irrigation. In Cuba, spent wash application at the rate of
90-150 m3 ha-1 increased the potassium content of the soil, with
increased cane yield and sugar recovery [13]. In a study
conducted in Kiev, Ukraine has shown increased yield of
grasses, maize and fodder beet by 45-100% using distillery
effluent. In India, extensive studies on distillery spent wash have
been carried out successfully with respect to various crops in
different agro-climatic regions [13]. Sugarcane (Saccharum
officinarum L.) has a worldwide significance as a major source of
food (sugar) and by-products which are economically more
important.
The present investigation aims to study the effect of
different concentration of distillery spent wash, inorganic fertilizer
and farmyard manure which are usually used by the local
farmers, on the growth and chlorophyll content of sugarcane
plant at Aska, Ganjam district, Odisha, India.

Corresponding Author, Email: [email protected], Tel.:- +919937576887

P.Rath et al./Rec Res Sci Tech 3 (2011) 169-176

Materials & Method
The field experiment was conducted with two different
doses of distillery spent wash (50% & 100%), fertilizer used by
the local farmers and fertilizer combined with organic manure
using Co 1274 sugarcane variety (Saccharum officinarum L.) as
test crop. The experimental field was divided into five sets of
three equal parts of size 5 m3 m each. Before plantation the
land was ploughed, labelled and divided into ridges and furrows
with uniform distance. The sugarcane test crop was collected
from the local farmer for planting. The setts were treated with
Bovistin and planted in the experimental field. After plantation
with regular watering once in a week the plants were allowed to
grow for three months. After a period of three months different
growth parameters and chlorophyll contents were recorded and

the fields were treated with different conditions (50 %, 100 %

effluent, fertilizer and fertilizer + cow dung). Out of the five sets
of fields with three replicates, one set was kept as control
without any treatment. Other four sets of fields were treated with
different conditions on every month and all the fields were
irrigated with water on every week. Biometric observations were
made randomly by selecting five plants in the net plot area of
individual treatments on every month.
Distillery spent wash application
The distillery spent wash samples were collected from the
Aska Co-operative Sugar Industries, Ltd., Ganjam district,
Odisha. The physicochemical properties of spent wash were
analyzed by standard methods of APHA [14] (Table 1).

Table 1- Chemical composition of DSW collected from Aska Co-operative Sugar Industries Ltd., Aska, Odisha.
Chemical parameters
pH
Electrical conductivity ( s)
Total solids
Total Dissolved Solids
Total Suspended Solids
Settleable Solids
Chemical Oxygen Demand(COD)
Biological Oxygen Demand(BOD)
Carbonate
Bicarbonate
Total Phosphorus
Total Potassium
Calcium
Magnesium
Sulphate
Sodium
Chlorides
Iron
Manganese
Zinc
Copper
Cadmium
Lead
Chromium
Nickel
Ammonical Nitrogen
Total Phosphorus
Total Potassium
Sulphur
Raw effluent was diluted to 50% v/v with water. The 50% and
concentrated (100%) effluents were taken for experimental
studies.
Fertilizer application
In this experiment the fertilizer doses used by the local
farmers were adopted for sugar cane crop. The inorganic
fertilizer was applied to the demarcated plots at four different
times i.e. NPK at the rate of 250 kg ha-1 before planting the setts;
urea at the rate of 125 kg ha-1 in two doses at 30 days and 90
days after plantation; the last dose of inorganic fertilizer, urea at
the rate of 60 kg ha-1+ Potash @ 62 kg ha-1 after 180 days of
plantation.

Amount (mg l-1)

7.23
28700
35340
27240
9980
9860
30520
15300
Negligible
12200
28.36
6500
920
753.25
5100
420
5626
6.3
1429
1.09
0.265
0.036
0.19
0.067
0.145
636.25
29.28
7300
75.6
Organic manure (cow dung)
In addition to the fertilizer dose the organic manure- cow
dung was applied to the experimental plots. The chemical
composition of cow dung is presented in Table 2. Total organic
carbon was measured by using the method by Nelson &
Sommers [15]. Total nitrogen was determined by Bremmer &
Mulvaney[16]. Total available phosphorus was determined by
using the method given by Bansal & Kapoor [17]. Total
potassium and calcium was determined by flame photometer
[17]. The pH and electrical conductivity (EC) were determined by
the method of Garg et al. [18]. Twenty kg of cow dung dissolved
and mixed in 20 litre of water and applied to the fields (plots)
over the fertilizers used.

P.Rath et al./Rec Res Sci Tech 3 (2011) 169-176

Table 2. Chemical composition of cow-dung used in the experiment at Aska,Ganjam, Odisha.
Chemical Parameters
kg-1)

Amount

Total Calcium (g
Mg (g kg-1)
Total Potassium (g kg-1)

1.29 0.3
0.194 0.26
4.69 0.068

Total Phosphorus (g kg-1)

3.9 0.04

Total Nitrogen(g

kg-1)

C:N ratio
Total organic carbon (g kg-1)

5.9 0.19
81.2 2.56
480.65 4.2

pH

7.6 0.03

Electrical conductivity (ds m-1)

1.96 0.02

Each value is the mean of five replicates SE

Soil analysis
The soil samples from the experimental site were collected,
air dried, powdered and analyzed for physico-chemical
properties. The physical properties like soil pH was estimated by

pH meter and chemical characteristics was determined by

following the methods of Chapman [19] and Wilde et al. [20] and
presented in Table 3.

Table 3. Properties of the soil of the experimental site, Aska, Ganjam (Odisha).
Nutrients

Amount (ppm)

pH (1:2) solution

6.9

Organic carbon (%)

8.97

Available nitrogen

345

Available phosphorus

131

Available potassium
Calcium

79
139

Magnesium
Sodium

226
84

Available sulphur

231

Iron
Manganese

192
199

Copper

04

The growth parameters such as height of the plant, length and

breadth of leaf, girth of the stem, number of leaves and number
of tillers per plant and leaf area were recorded as per the
standard procedure. From leaf area leaf area index was
calculated [21]. The chlorophyll content of the sugarcane plant
was determined by using method of Arnon [22] and expressed
as mg g-1 fresh wt.
Data analysis
Data relating to different growth parameters and total
chlorophyll content was analyzed by one way ANOVA. Results
of different conditions were computed over control and their
difference among treatments was tested through least significant
difference (LSD). All the parameters are statistically significant.
Results and discussion
Distillery spent wash is the unwanted liquid waste produced
during the production of alcohol and it is one of the most
important environmental issues. The distillery spent wash with
its characteristic unpleasant odor poses a serious threat to the
water quality around the world [23]. The ever increasing amount
of distillery spent wash and its disposal has stimulated the need
for developing new technologies to process this effluent

efficiently and economically including growth and yield of

different crops in agriculture [24]. It was found that the growth
and chlorophyll content of the sugarcane in different conditions
of treatment showed an increasing trend over the control.
Average height of the sugarcane plant after 210 days showed
an increase of 13.45% in the 50% DSW treated plot over the
control (Fig. 1). However, the growth showed negative trend in
100% DSW.) in all parameters. The average length of leaves of
the test crop after 210 days of plantation showed an increase of
11.22% in 50% DSW treated plants over control and a negative
trend in 100% DSW (Fig. 2). The average breadth of the leaves
after 210 days of plantation showed an increase of 46.96% in
the 50% DSW plants over control (Fig. 3) and a negative trend
in 100% DSW.
Similarly the average girth of the stem, average number of
leaves per plant, average number of tillers per plant, average
leaf area index and chlorophyll content of the test crop after 210
days of plantation showed an increase of 16.79%, 21.18%,
57.5%, 62.84% and 28.99% respectively in 50% DSW treated
plants over control (Fig. 4, 5, 6, 7, 8) and a negative trend in
100% DSW. The ANOVA showed that there was significant
difference in all the stages of growth of the plant except in 90

P.Rath et al./Rec Res Sci Tech 3 (2011) 169-176

days (Table 4, 5,6,7,8,9,10, & 11). All fertilizer and fertilizer+cow

dung treated plants showed an increasing trend over control.

Fig. 1. Height of the sugarcane plant at different growth periods with different treatments.

Table 4. Statistical analysis of the height of the sugarcane plant at different days of growth and different treatments
Days after plantation
90
120
150
180
210

F value for different

Significance level
treatments
0.54
NS
4.62
0.05p
3.32
0.05p
3.45
0.05p
3.99
0.05p
NS Not Significant

LSD
23.96
23.43
22.92
19.88
28.89

Fig. 2 . Length of leaves of sugarcane plant at different growth periods with different treatments.

Table 5. Statistical analysis of the length of leaves of sugarcane plant at different days of growth and different treatments.
Days After Plantation
90
120
150
180
210

F value for different

treatments
0.46
3.04
5.27
3.17
6.1

Significance level

LSD

NS
0.05p
0.05p
0.05p
0.05p

24.46
12.98
12.40
18.64
11.01

NS Not Significant

P.Rath et al./Rec Res Sci Tech 3 (2011) 169-176

Fig. 3. Breadth of leaves of sugarcane plant at different growth periods with different treatments.

Table 6. Statistical analysis of breadth of leaves of the sugarcane plant at different days of growth and different treatments
Days after plantation
F value for different
Significance level
LSD
treatments
90
0.88
NS
0.67
120
4.4
0.05p
0.85
150
8.02
0.05p
0.89
180
7.8
0.05p
0.83
210
16.5
0.05p
0.76
NS Not Significant

Fig. 4. Girth of the stem of sugarcane plant at different growth period with different treatment

Table 7. Statistical analysis of the girth of the stem of the sugarcane plant at different days of growth and different treatment
Days after plantation
F value for different
Significance level
LSD
treatments
90
1.16
NS
0.83
120
4.97
0.05p
0.78
150
7.68
0.05p
0.47
180
4.42
0.05p
0.50
210
6.14
0.05p
0.70
NS Not Significant

Ritesh Vernekar et al./Rec Res Sci Tech 3 (2011) 169-176

Figure .5. Number of leaves per sugarcane plant at different growth periods with different treatments

Table 8. Statistical analysis of number of leaves per sugarcane plant at different days of growth and different treatment
Days after plantation
F value for different
Significance level
LSD
treatments
90
0.184
NS
2.09
120
11.57
0.05p
2.15
150
7.02
0.05p
1.82
180
4.36
0.05p
2.33
210
6.14
0.05p
2.04
NS - Not significant
Fig. 6. Numbers of tillers per sugarcane plant at different growth periods with different treatments

Table 9 .Statistical analysis of number of tillers per sugarcane plant at different days of growth and different treatment.
Days after plantation
F value for different
Significance level
LSD
treatments
90
0.45
NS
1.15
120
3.49
0.05p
2.70
150
7.02
0.05p
2.13
180
3.29
0.05p
3.49
210
7.9
0.05p
2.10
NS Not Significant

P.Rath et al./Rec Res Sci Tech 3 (2011) 169-176

Figure. 7. Leaf area index of sugarcane crop at different growth periods with different treatments

analysis Table 10. Statistical of leaf area index of sugarcane plant at different days of growth and different treatment
Days after plantation
F value for different
Significance level
LSD
treatments
90
0.63
NS
2.11
120
6.51
0.05p
2.03
150
12.07
0.05p
2.06
180
15.55
0.05p
1.88
210
25.83
0.05p
1.74
NS Not Significant
Fig. 8. Chlorophyll content of sugarcane leaf at different growth periods with different treatments

Table11.Statistical analysis of chlorophyll content of leaves of of sugarcane plant at different days of growth and different treatment
Days after plantation
F value for different
Significance level
LSD
treatments
90
0.39
NS
0.57
120
4.02
0.05p
1.27
150
5.29
0.05p
1.17
180
3.28
0.05p
1.21
210
3.18
0.05p
1.26
NS Not Significant
The spent wash treated soil is enriched with the plant nutrients
such as nitrogen, phosphorus and potassium. Subsequent use
of spent wash for irrigation enriches the soil fertility without any
adverse effect and hence spent wash (50%) can be
conveniently used for the irrigation of sugarcane crop without
any external (either organic or inorganic) fertilizer application.
Spent wash increases the soil enzymatic activity [25] which
enriches the plant growth and development. The declining
tendency of growth parameters in higher concentration may be

due to the presence of higher amount of organic matter and

biological oxygen demand (BOD) which might have lead to
depletion of O2 and accumulation of CO2 in the soil.
The increase in chlorophyll content in 50% distillery spent wash
treatment crop suggests that synthesis of chlorophyll is
accelerated in low concentration. Similar findings have been
reported by many workers [26, 27, 28]. The increase in
chlorophyll content may be due to lack of heavy metals and in

P.Rath et al./Rec Res Sci Tech 3 (2011) 169-176

the effluent and probably the availability of Fe and Mg which are
necessary for the synthesis of chlorophyll. Reduction of
chlorophyll pigment in higher concentration (100%) may be
associated with mineral ions. Some of the possible reasons for
the decrease of pigment contents may be due to the formation
of enzyme chlorophyllase which is responsible for chlorophyll
degradation [29, 30].
Conclusion
From the experimental observations it may be concluded that
distillery spent wash at 50% v/v concentration may serve as a
good liquid fertilizer for sugarcane crop which is more effective
than that of the fertilizer used by the local farmers as well as the
inorganic fertilizer plus the organic manure ( cow dung ).

12.
13.
14.
15.

16.
Acknowledgements
Authors are thankful to the Aska Co-operative Sugar Industries
Ltd and P.K. Panda, Principal, Aska Science College for
providing necessary facilities. We are thankful to the sugarcane
growers who helped during the experimental study. University
Grants Commission, New Delhi is thankfully acknowledged for
providing financial assistance in the form of a minor research
project to the senior author.
References
1. Kuntal, M. H., A. K. Biswal, K. Bandyopadhyaya and K.
Mishra. 2004. Effect of post methanation effluent on soil
physical properties under a Soyabean-Wheat system in a
vertisol. J. Plant Nutri. & Soil Sci. 167( 5):584-590
2. Chakarbarty, R. N. 1980. Management of sugar and
alcohol industry wastes- pollution control and conservation
of energy and fertilizer. Proc. Symp. Disposal of sugar and
distillery waste, U. P. Water Pollution Control Board,
Lucknow.
3. Suganya, K., Rajannan, G. 2009. Effect of one time postsown and pre-sown application of distillery spent wash on
the growth and yield of maize crop. Botany Research
International 2(4):288-294.
4. Diangan Jr., M. Perez, T. and Claveria, R. 2008. Analysis
of land application as a method of disposal of distillery
effluent Int. J. Env. Health 2: 258- 271.
5. Swaminathan, K. and Vaidheeswaran, P. 1991. Effect of
dyeing factory effluent on seed germination and seedling
development of groundnut (Arachis hypogea). J. Env.
Biol. 12(3): 253- 258.
6. Rani, R. and Srivastata, M. M. 1990. Ecophysiological
responses of Pisum sativum and Citrus maxima to distillery
effluents. Int. J. Eco. Environ. Sci. 16-23.
7. Badawy, S. H. and Elmataium, R. A. 1999. Effect of
irradiated and non-irradiated sewage sludge application on
some nutrient-heavy metal content of soil and tomato plant.
1st Congress on Recent Technology on Agriculture.
Bulletin Faculty Agriculture. University of Cairo, Cairo. pp.
728-744.
8. Valdes, E., Obaya, M, C. and Ramos, J. 1996. Ecology
and the Sugar Industry. Revista 30:214-229.
9. Shomasheka, R. K. and Siddaramaiah, K. 1993. Effect of
soap and detergents factory effluent on soils and crop
plants. J. Ind. Pollut 9(2):77-84
10. Dutta, S. K. and Boissya, C. L. 1997. Effect of paper mill
effluent on germination of rice (Oryza sativa. L var. masun )
and growth behaviour of its seedlings. J. Ind. Pollut. 13 (1):
41-47.
11. Karunyal, S., Renuga, G. and Paliwal, K. 1993. Effect of
tannery effluent on seed germination, leaf area, biomass

17.
18.

19.
20.
21.
22.
23.

24.
25.
26.
27.

28.

29.

30.

and mineral content of some plants. Bioresource Technol

47: 218-218.
Mathur, A. C., Davis, J. B.1987. Palm oil mill effluent. A
review of methods proposed for its treatment. Trop. Sci.
2:233-262
Kanimozhi, R. and Vasudevan, N. 2010. An over view of
waste water treatment in distillery industry. Int. J. Env.
Engineering 2:159-184
APHA, 1992. Standard Methods for Examination of Waste
Water Analysis. 17th ed., Washington.
Nelson, D. W. and Sommers, L. E. 1982. Total carbon and
organic carbon and organic matter. In: A. L. Page, R. H.
Miller and D. R. Keeney (eds.). Method of Soil Analysis.
American Society of Agronomy, Meddison, 11: 539-579
Bremmer, J. M, and Mulvarey, R. G. 1982. Nitrogen Total.
In: A. L Page, R.H. Miller and D. R. Keeney, (eds.). Method
of Soil Analysis, American Society of Agronomy Madison.
pp. 575-624.
Bansan, S. and Kapoor, K. K. 2000. Vermicomposting of
crop residues and cattle dung with Eisenia foetida.
Bioresources Technol. 73:95-98.
Garg, V. K., Yadav, Y. K., Sheoran, Chand, S. & Kausik, P.
2006. Live stocks excreta management through
vermicomoposting using an epigeic earthworm Eisenia
foetida. Environmentalist 26:269-276.
Chapman, S, B (ed.).1976. Methods in Plant Ecology,
Oxford: Black Well Scientific Publication. pp. 411- 466.
Wild, S. A., R. B. Coreg, J. S. Iyer & G. K. Voigt. 1979. Soil
and Plant Analysis for Tree Culture (5th ed ) Oxford & IBH.,
New Delhi
Kemp, C. D. 1960. Methods of estimating the leaf area of
grasses from linear measurements. Ann. Bot. (N.S.) 24:
4491 4499.
Arnon D. I. 1994. Copper enzymes in isolated chloroplasts,
polyphenol oxidase in Beta vulgaris. Plant Physiol 24: 115.
Joshi, H. C., Karla, N., Choudhury, A. and Deb, D. L. 1994.
Environmental issues related with distillery effluent
utilization in agriculture in India. Asia Pacific Journal of
Environment Development 1:92 103.
Sarayu Mohana, Bhavik, K, Acharya and Datta Madamwar.
2009. Distillery spent wash treatment technologies and
potential applications. J. Hazardous Material 163(1):12-25.
Kalaiselvi, P., Mahimairaja, S. 2009. Effect of
biomethanated spent wash on soil enzymatic activities.
Botany Research International. 2(4): 267-272.
Srivastava, N. and Sahai, R. 1987. Effect of distillery waste
in the performance of Cicer arietinum L. Environ. Pollut.
43. 91-102.
Kurunyal, S, Renuha, G, and Palliwal, K. 1993. Effect of
tannery on the seed germination, leaf area, biomass and
mineral content of some plants. Bioresource Technol. 47:
215-218.
Orhue, E. R., Osaigbova, A. U. and Vwioko, D. E. 2005.
Growth of maize (Zea mays L.) and changes in some
chemical properties of an Utisol amended with brewery
effluent. Afri. J. Biotechnol. 4(9): 973-978.
Majumdar, S., S. Ghose, B. R. Glick and Dumbroff, 1991.
Activities of chlorophyllase, phosphoenol pyruvate
carboxylase and ribulose 1, 5-bi-phosphate-carboxylase in
the primary leaves of Soyabean during senescence and
drought. Physiol. Plant 81: 473-480.
Krishna, K and Leelavathi. 2002. Toxicity of sugar factory
effluent to germination, vigour index and chlorophyll
content of paddy. Nature Environ. Pollut. Tech., 1(3): 249253.