The effects of sugar mill effluent on hybrid cultivar of Faba Bean (Vicia faba L.) and soil properties CO 3 -2

Abstract

In this study, the ferti-irrigational response of hybrid cultivar of Vicia faba (var. VH-82-1) with sugar mill effluent was investigated. Five different doses of sugar mill effluent (20, 40, 60, 80 and 100%) were used for ferti-irrigation of V. faba along with borewell water as control. The study revealed that sugar mill effluent had significant (P<0.01) effect on EC, pH, OC, HCO 3 -, of the soil. Insignificant (P>0.05) changes in moisture content, WHC and bulk density of the soil were observed after ferti-irrigation with sugar mill effluent. The agronomic parameters namely shoot length, root length, number of flowers, pods, dry weight, chlorophyll content, leaf area index (LAI), crop yield and harvest index (HI) of V. faba were gradually increased from 20 to 40% and decreased from 60 to 100% concentration of sugar mill effluent as compared to the control. The biochemical components namely crude proteins, crude fiber and total carbohydrates were noted maximum with 40% concentration of sugar mill effluent. The contamination factor of heavy metals was in order of Cr>Zn>Cd>Mn>Cu for soil and Cr>Cu>Cd>Mn>Zn for V. faba after fertigation with sugar mill effluent. The translocation of various heavy metals in different parts of V. faba were in order of leaves>shoot>root> fruits for Cu, Mn and Zn; root>shoot>leaves>fruit for Cd; and shoot>root>leaves>fruit for Cr after sugar mill effluent irrigation. Thus, sugar mill effluent can be used with appropriate dilution for the maximum yield of V. faba.

Full text

International Journal of Biotechnology Research Vol. 1(6), pp. 091-102, July 2013
Available online at http://academeresearchjournals.org/journal/ijbr
ISSN 2328-3505 ©2013 Academe Research Journals
Full Length Research Paper
The effects of sugar mill effluent on hybrid cultivar of
Faba Bean (Vicia faba L.) and soil properties
Vinod Kumar* and A.K. Chopra
Department of Zoology and Environmental Science, Faculty of Life Sciences, Gurukula Kangri University,
Haridwar-249404 (Uttarakhand), India.
Accepted 10 July, 2013
In this study, the ferti-irrigational response of hybrid cultivar of Vicia faba (var. VH-82-1) with sugar mill
effluent was investigated. Five different doses of sugar mill effluent (20, 40, 60, 80 and 100%) were used
for ferti-irrigation of V. faba along with borewell water as control. The study revealed that sugar mill
effluent had significant (P<0.01) effect on EC, pH, OC, HCO3-, CO3-2, Na+, K+, Ca2+, Mg2+, TKN, PO43-, SO42-,
Fe2+, Cd, Cr, Cu, Mn and Zn of the soil. Insignificant (P>0.05) changes in moisture content, WHC and
bulk density of the soil were observed after ferti-irrigation with sugar mill effluent. The agronomic
parameters namely shoot length, root length, number of flowers, pods, dry weight, chlorophyll content,
leaf area index (LAI), crop yield and harvest index (HI) of V. faba were gradually increased from 20 to
40% and decreased from 60 to 100% concentration of sugar mill effluent as compared to the control.
The biochemical components namely crude proteins, crude fiber and total carbohydrates were noted
maximum with 40% concentration of sugar mill effluent. The contamination factor of heavy metals was
in order of Cr>Zn>Cd>Mn>Cu for soil and Cr>Cu>Cd>Mn>Zn for V. faba after fertigation with sugar mill
effluent. The translocation of various heavy metals in different parts of V. faba were in order of
leaves>shoot>root> fruits for Cu, Mn and Zn; root>shoot>leaves>fruit for Cd; and
shoot>root>leaves>fruit for Cr after sugar mill effluent irrigation. Thus, sugar mill effluent can be used
with appropriate dilution for the maximum yield of V. faba.
Key words: Sugar mill effluent, Vicia faba, irrigation, agronomical practices, heavy metals, contamination
factor.
INTRODUCTION
Irrigation with wastewater is a common practice in urban
and suburban areas (Rath et al., 2011; Kumar and
Chopra, 2012). Effluent generated from various sources
like municipal, household, small and big industries are
the important sources of wastewater generation (Ramana
et al., 2002; Nath et al., 2007; Kumar, 2010). Use of
wastewater in agriculture reduces the irrigation water cost
as it is available without paying any cost, and also
reduces the fertilizer cost as it is rich in various nutrients
required for plant growth (Bharagava et al., 2008; Kannan
and Upreti, 2008). Besides this, effluent contains heavy
metals which accumulates in plant and vegetable parts,
and cause various health effects (Hati et al., 2007;
Pandey et al., 2008). Long term irrigation with effluents
increases organic carbon and heavy metals accumulation
in soil, and increase the chances of their entrance in food
chain, and this ultimately causes significant
geoaccumulation, bioaccumulation and biomagnifications
(Muchuweti et al., 2006; Chopra et al., 2009). The nature
of the soil is one of the most important factors in
determining the heavy metal content of food plants
(Itanna, 2002). However, the heavy metals content in
plants can also be affected by other factors such as the
*Corresponding author. E-mail: drvksorwal@gmail.com. Tel:
+91-1334-249091.
Abbreviations: BD, Bulk density; LAI, Leaf area index; HI,
Harvest index; RT, Relative toxicity; WHC, Water holding
capacity.

Kumar and Chopra 092
application of fertilizers, sewage sludge or irrigation with
wastewater (Frost and Ketchum, 2000; Muchuweti et al.,
2006). Presently, India has nearly 650 sugar mill that
produce about 15 million tons of sugar and 13 million
tons of molasses (Baskaran et al., 2009). Sugar mills
account in the industries which discharge huge amount of
effluent per day without any or partial treatment during
the crushing season (Baruah et al., 1993; Ezhilvannan et
al., 2011). Sugar mill effluent contains high magnitude of
pollution load, and caused adverse effect on soil and
biological system (Arindam and Prasad, 1999).
Vicia faba (Faba bean) is cultivated in winter in India. It
is extensively used as green vegetable and its seeds as
pulse and also has medicinal values (Singh et al., 2012;
Gutierrez et al., 2006). The green pods are mildly diuretic
and contain a substance that reduces the blood sugar
level (Martin et al., 1991; Singh et al., 2012). The dried
mature pod is used in the treatment of diabetes and in
the treatment of ulcers (Sprent et al., 1977; Bond et al.,
1985). Most crops give higher potential yields with
wastewater irrigation; reduce the need for chemical
fertilizers, resulting in net cost savings to farmers. So it is
important to understand the specificity of crop-effluent
liaison for their appropriate application in irrigation
practices (Kumar, 2010). In recent past, various studies
have been carried out on the characteristics of effluent of
industries, and the agronomical properties of various crop
plants (Ramana et al., 2002; Hati et al., 2007; Bharagava
et al., 2008; Kannan and Upreti, 2008; Kumar, 2010). But
much attention has not been paid so far on the use of
industrial effluents on the cultivation of agricultural crops
like V. faba. Keeping in view the reuse of wastewater as
irrigant and the economic importance of V. faba, the
present investigation was undertaken to study the ferti-
irrigation response of hybrid cultivar of faba bean (Vicia
faba L.) with sugar mill effluent.
MATERIALS AND METHODS
Experimental design
A field study was conducted in the Experimental Garden
of the Department of Zoology and Environmental
Sciences, Faculty of Life Sciences, Gurukula Kangri
University Haridwar, India (29°55'10.81'' N and 78°96
07'08.12'' E), during the year 2011 and 2012. Eighteen
plots (each plot had an area of 9 m2) were selected for six
treatments of sugar mill effluent namely 0% (control), 20,
40, 60, 80 and 100% for the cultivation of V. faba. The
experiment was conducted under completely randomized
designed and replicated three times.
Effluent collection and analysis
R.B.N.S. sugar mill Laksar, Haridwar (29°44'46"N
78°1'46"E) was selected for the collection of effluent
samples. The treated effluents were collected from outlet
of the effluent treatment plant situated in the campus of
the sugar mill. The effluent was brought to the laboratory
and analyzed for various physico-chemical and
microbiological parameters namely: TDS, pH, electrical
conductivity (EC), dissolved oxygen (DO), biochemical
oxygen demand (BOD), chemical oxygen demand
(COD), chlorides (Cl-), bicarbonates (HCO3-), carbonates
(CO3-2), sodium (Na+), potassium (K+), calcium (Ca2+),
magnesium (Mg2+), total Kjeldahl nitrogen (TKN), nitrate
(NO32-), phosphate (PO43-), iron (Fe2+), cadmium (Cd),
chromium (Cr), copper (Cu), manganese (Mn), zinc (Zn),
standard plate count (SPC) and most probable number
(MPN) following standard methods (APHA, 2005) and
further used for fertigation of V. faba.
Sowing of seeds, irrigation pattern and soil analysis
The seeds of V. faba (var. VH-82-1) were procured from
Indian council of agriculture research (ICAR), Pusa, New
Delhi and sterilized with 0.01% mercuric chloride and
were soaked for 12 h. Seeds were sown in 10 rows with a
distance of 30.0 cm between rows, while distance
between the seeds was 15 cm. The thinning was done
manually after 15 days of germination to maintain the
desired plant spacing and to avoid competition between
plants. Plants in each plot were fertigated twice in a
month with 50 gallons of sugar mill effluent with 20, 40,
60, 80 and 100% treatments along with bore well water
as the control. The soil was analyzed before sowing and
after harvesting of crop for various physico-chemical
parameters namely: bulk density (BD), water holding
capacity (WHC), soil texture, EC, pH , Cl-, OC, Na+, K+,
Ca2+, Mg2+, Fe2+, TKN, PO43- and SO42-, Cd, Cr, Cu, Mn
and Zn using standard methods (Chaturvedi and Sankar,
2006).
Study of crop parameters
Agronomical parameters of V. faba from germination to
maturity (0-90 days) were determined following standard
methods (Chandrasekar et al., 1998 for seed
germination, relative toxicity, shoot length, root length,
number of flowers number of pods and crop yield),
(Milner and Hughes, 1968 for dry weight), (Porra, 2002
for chlorophyll content), (Denison and Russotti, 1997 for
leaf area index, LAI) and (Sinclair, 1998 for Harvest
index, HI). The nutrients quality of V. faba were
determined by using the following parameters: crude
protein (4.204 AOAC, 1980), crude fiber (4.601 AOAC,
1980) and the total carbohydrate in dry matter were
determined by the anthrone reagent method (Cerning
and Guilhot, 1973).
Heavy metals analysis
For heavy metal extraction, 10 ml sample of sugar mill
effluent, and 1.0 g of air dried soil or plants were taken in

Int. J. Biotechnol. Res. 093
Table 1. Physico-chemical and microbiological characteristics of control (Bore well water) and R.B.N.S. sugar mill effluent.
Parameter
Borewell water
(Control )
Effluent concentration (%)
BISa for
irrigation water
20
40
60
80
100
TDS (mg L -1)
278.50±10.75
1285.00±5.77
1898.00±3.65
2542.00±3.83
3625.50±4.43
4868.00±12.65
1900
EC (dS cm-1)
0.43±19
2.26±0.27
3.44±0.40
4.76±0.44
6.83±0.27
8.65±0.86
-
pH
7.46±0.24
7.64±10
7.80±0.09
8.26±10
8.85±17
8.98±0.41
5.5-9.0
BOD (mg L -1)
2.86±0.66
182.64±3.21
414.63±4.44
824.70±3.43
1228.55±7.79
1664.56±5.97
100
COD (mg L -1)
5.88±1.37
231.08±4.29
569.50±4.43
1134.25±7.93
1697.50±10.50
2285.80±7.75
250
Cl - (mg L -1)
15.68±2.50
138.34±5.88
331.80±2.61
648.32±3.02
940.31±4.64
1275.42±11.62
500
HCO3- (mg L -1)
282.00±13.95
319.97±3.15
337.11±5.89
372.74±4.21
503.37±8.37
659.00±12.49
-b
CO3 -2 (mg L -1)
105.75±5.91
130.81±3.18
166.03±6.93
173.57±4.63
198.91±8.41
229.75±5.06
-
Na+ (mg L -1)
9.65±1.25
33.61±2.83
78.74±3.32
136.61±5.04
203.67±8.97
251.50±12.04
-
K+ (mg L -1)
5.54±2.25
39.53±2.44
93.14±4.51
177.37±4.05
259.83±3.39
336.00±7.83
-
Ca2+ (mg L -1)
45.76±4.16
108.07±4.61
243.24±3.56
442.84±4.29
650.35±1.56
884.20±9.31
200
Mg2+ (mg L -1)
12.15±1.50
40.97±2.38
82.62±2.85
121.33±8.61
166.53±3.47
243.50±7.19
-
TKN (mg L -1)
1.45±5.08
44.72±2.03
89.75±3.23
132.43±3.61
173.60±4.61
231.45±9.93
100
PO43- (mg L -1)
0.06±0.01
35.67±2.58
74.66±2.59
148.71±3.77
224.45±2.50
296.82±9.32
-
SO42- (mg L -1)
67.22±2.34
110.21±4.40
256.38±2.53
468.32±2.45
684.53±4.37
893.76±9.18
1000
Fe2+ (mg L -1)
0.35±0.06
4.98±1.75
7.38±2.80
15.18±2.77
18.85±2.07
26.58±4.27
1.0
Cd (mg L -1)
BDLc
0.74±0.02
3.87±0.11
7.20±0.12
10.36±0.16
14.96±0.13
15
Cr (mg L -1)
BDL
0.29±0.03
0.74±0.04
1.84±0.10
3.24±0.25
7.86±0.06
2.00
Cu (mg L -1)
0.10±0.02
0.34±0.01
1.38±0.1
2.05±0.03
3.11±0.03
4.56±0.14
3.00
Zn (mg L -1)
0.45±0.03
2.13±0.26
3.72±0.37
7.17±0.81
9.72±0.42
12.63±1.54
15
Mn ( mg L -1)
0.16±0.01
1.12±0.08
2.65±0.13
4.23±0.24
6.56±0.44
8.97±0.96
1.00
SPC (SPC ml -1)
4.53×102±12
6.74×104±48
6.54×105±56
9.86×106±98
4.85×107±78
9.45×108±86
10000
MPN (MPN100 ml -1)
3.29 ×101±15
6.88×103±34
7.55×103±42
5.98×104±65
8.43×105±86
7.74×106±100
5000
Mean ± of four values; aBIS - Bureau of Indian standard; b- Not defined in standard; cBDL - Below detection limit.
digestion tubes with 3 ml of concentrate HNO3 and
digested in an electrically heated block for 1 h at 145°C. 4
ml of HClO4 was added to this mixture and heated to
240°C for 1 h. The mixture was cooled and filtered
through Whatman # 42 filter paper and aliquot was made
to 50 ml with double distilled water and used for analysis.
Heavy metals were analyzed using an atomic absorption
spectrophotometer (PerkinElmer-800) following methods
of Chaturvedi and Sankar (2006). The contamination
factor for heavy metals accumulated in sugar mill effluent
irrigated soil and V. faba was calculated following
Håkanson (1980).
Statistical analysis
Data were subjected to two-way ANOVA by using SPSS
(ver. 12.0, SPSS Inc., Chicago, Ill). Duncan’s multiple
range test was also performed to determine the
difference between applications that were significant or
non significant. Mean standard deviation and coefficient
of correlation (r-value) of soil and crop parameters with
effluent concentrations were calculated with MS Excel
(ver. 2003, Microsoft Redmond Campus, Redmond, WA)
and graphs were plotted with Sigma plot (ver. 12.3,
Systat Software, Inc., Chicago, IL).
RESULTS AND DISCUSSION
Characteristics of effluent
The mean±SD values of physico-chemical and
microbiological parameters of sugar mill effluent are
given in Table 1. The results revealed that it was
yellowish in color with odor of sugar, alkaline in nature
having pH (8.98). The alkaline nature of the sugar mill
effluent might be due to the presence of high
concentrations of alkalis used in sugar production.
Among various parameters of effluent (100%), TDS
(4868 mg L-1), BOD (1664.56 mg L-1), COD (2285.80 mg
L-1), Cl – (1275.42 mg L-1), TKN (231.45 mg L-1), Ca2+
(884.20 mg L-1), Fe (26.58 mg L-1), Zn (12.63 mg L-1), Cu
(4.56 mg L-1), Mn (8.97 mg L-1), MPN (7.74×106 MPN100
ml-1), and SPC (9.45×108 SPC ml-1) were found beyond
the prescribed limit of Indian irrigation standards (BIS,
1991). High BOD and COD might be due to the presence
of high utilizable organic matter and rapid consumption of
dissolved inorganic materials (Kumar, 2010). The higher
bacterial load (SPC and MPN) in sugar mill effluent might

Kumar and Chopra 094
Table 2. Physico-chemical characteristics of soil after irrigation with R.B.N.S. sugar mill effluent.
Parameter
Borewell
water
(Control)
Effluent concentrations (%)
F-
calculated
CD
20
40
60
80
100
Soil moisture (%)
67.38±5.21
65.42±4.16
64.33±3.76
63.27±4.10
61.45±4.69
60.54±3.27
3.22 NS
4.78
WHC (%)
46.23±3.44
45.84±3.01
44.61±2.69
43.28±2.99
42.52±3.43
41.57±4.32
1.74 NS
5.01
BD (gm cm-3)
1.45±0.18
1.44±0.15
1.43±0.12
1.41±0.13
1.39±0.13
1.39±0.16
0.41 NS
0.12
pH
7.55±0.30
7.74±0.41
7.83±0.40
8.10a±0.37
8.16a±0.17
8.28a±0.22
3.54*
0.45
EC (dS m-1)
2.07±0.10
2.85a±0.10
3.00a±0.10
3.89a±0.13
4.36a±0.09
5.40a±0.13
52.34**
0.18
OC (mg Kg -1)
0.45±0.08
4.88a±0.13
7.77a±0.64
9.46a±0.83
12.59a±1.82
15.98a±1.64
55.43**
1.50
HCO3- (mg Kg -1)
293.64±4.16
346.58a±6.69
384.62a±3.02
441.64a±6.71
463.97a±5.88
498.36a±4.82
123.29**
7.64
CO3-2 (mg Kg -1)
229.65±5.45
238.30a±3.02
243.96a±4.29
271.08a±5.76
283.41a±3.90
297.12a±8.22
108.92**
7.57
Na+ (mg Kg -1)
18.81±2.48
30.72a±3.99
32.75a±3.79
40.71a±4.71
45.23a±3.45
53.70a±4.59
49.35**
5.13
K+ (mg Kg -1)
155.34±4.26
205.25a±4.06
213.88a±2.56
221.33a±3.69
231.07a±3.75
238.29a±6.58
120.67**
7.3
Ca2+ (mg Kg -1)
15.36±4.73
32.81a±3.20
69.72a±3.60
131.56a±4.36
170.62a±3.43
234.24a±4.32
108.14**
6.79
Mg2+ (mg Kg -1)
1.70±0.48
5.46a±0.62
12.47a±1.47
16.84a±1.21
24.61a±2.12
33.61a±3.08
145.36**
2.74
TKN (mg Kg -1)
32.21±3.34
86.28a±4.59
156.75a±4.45
266.92a±3.47
347.28a±4.34
390.78a±6.70
448.74**
6.59
PO43- (mg Kg -1)
53.00±2.58
62.63a±3.44
117.68a±3.47
152.29a±5.17
216.75a±6.49
278.10a±3.39
474.58**
6.39
SO42- (mg Kg -1)
74.37±2.07
84.20a±4.80
97.41a±3.80
123.60a±3.57
189.02a±4.84
237.98a±6.60
91.01**
7.04
Fe2+ (mg Kg -1)
2.65±0.81
4.23a±0.16
5.39a±0.27
7.60a±0.24
8.31a±0.14
9.86a±0.73
91.91**
0.76
Cd (mg Kg -1)
0.55±0.03
1.58a±0.05
2.87a±0.11
3.75a±0.13
5.78a±0.12
6.89a±0.11
39.63***
0.53
Cr (mg Kg -1)
0.32±0.02
0.59a±0.01
0.96a±0.06
1.95a±0.08
3.45a±0.07
4.98a±0.23
3.03***
0.018
Cu (mg Kg -1)
0.77±0.34
1.76a±0.39
1.98a±0.56
2.88a±0.41
4.75a±0.77
5.89a±0.57
166.45**
0.73
Mn (mg Kg -1)
0.79±0.13
1.97a±0.11
2.36a±0.20
2.97a±0.11
4.55a±0.13
6.98a±0.31
144.98**
0.25
Zn (mg Kg -1)
0.85±0.04
2.64a±0.05
3.13a±0.09
6.28a±0.05
8.27a±0.05
10.78a±0.03
13.87**
0.08
Mean ± of four values; Significant F - **P>1% level, *P>5% level; NS - Not Significant; CD - Critical difference.
be due to the presence of more dissolved solids and
organic matter in effluent as earlier reported by Kumar
and Chopra (2012). The alkaline pH (8.05) and higher
total solids (2395.00 mg L-1), EC (12.8 dS m-1), Na
(3200.00 mg L-1) and COD (142.00 mg L-1) indicated that
there is higher inorganic and organic load in the sugar
mill effluent of Co-operative Sugar Mill, Rohtak, Haryana,
India, as reported by Kaushik et al. (1996).
Characteristics of soil
The mean ± SD values of various physico-chemical
characteristics are given in Table 2. During the present
study, no change in soil texture occurred with the
application of all the concentrations of sugar mill effluent
throughout the period of the experiment. The soil
characteristics changed on irrigation with different
concentrations (20 to 100%) of the effluent. Among
different concentrations of sugar mill effluent, irrigation
with 100% effluent concentration showed maximum
decrease in moisture content, WHC, BD, increase in EC,
OC, HCO3-, CO3-2, Na+, K+, Ca2+, Mg2+, TKN, PO43-, SO42-
, Fe2+, Cd, Cr, Cu, Mn and Zn of the soil.
The ANOVA analysis indicated that sugar mill effluent
showed insignificant (P>0.05) effect on moisture content,
WHC and BD of the soil. All concentrations (20 to 100%)
of sugar mill effluent showed significant (P<0.01) effect
on EC, OC, HCO3-, CO3-2, Na+, K+, Ca2+, Mg2+, TKN,
PO43-, SO42-, Fe2+, Cd, Cr, Cu, Mn and Zn in the V. faba
cultivated soil (Table 2). pH of the soil was found to be
significantly (P<0.05) different with sugar mill effluent
concentrations. The r value of the soil parameters
namely: pH (r = +0.93), EC (r = +0.98), OC (r = +0.92),
HCO3- (r = +0.90), CO3-2 (r = +0.96), Na+ (r = +0.94), K+ (r
= +0.73), Ca2+ (r = +0.97), Mg2+(r = +0.90), TKN (r =
+0.97), PO43- (r = +0.85), SO42- (r = +0.95) Fe2+ (r =
+0.93), Cd (r = +0.88), Cr (r = +0.86), Cu (r = +0.83), Mn
(r = +0.85) and Zn (r = +0.80) showed their positive
correlation with different concentrations of sugar mill
effluent (Table 3). Moisture content (r = -0.89), BD (r = -
0.92) and WHC (r = -0.84) of the soil were recorded to be
negatively correlated with sugar mill effluent
concentrations (Table 3). The concentration of heavy
metals, Cd, Cr, Cu, Mn and Zn in the soil were increased
as per the effluent concentration increased (Table 2). The
contamination factor indicated the contamination rate of
heavy metals in the soil after sugar mill effluent irrigation.
The contamination factor of various heavy metals in the
soil was in order of Cr>Zn>Cd>Mn>Cu after irrigation
with sugar mill effluent (Figure 1). Total average organic
carbon in the soil irrigated with sugar mill effluent was
higher than the soil irrigated with bore well water. The

Int. J. Biotechnol. Res. 095
Table 3. Pearson correlation (r) value of R.B.N.S. sugar mill
effluents to soil characteristics.
Effluent concentration/soil characteristics
r value
Sugar mill effluent versus soil moisture content
-0.89
Sugar mill effluent versus soil WHC
-0.84
Sugar mill effluent versus soil BD
-0.92
Sugar mill effluent versus soil pH
0.93
Sugar mill effluent versus soil EC
0.98
Sugar mill effluent versus soil OC
0.92
Sugar mill effluent versus soil HCO3-
0.90
Sugar mill effluent versus soil CO3-2
0.96
Sugar mill effluent versus soil Na+
0.94
Sugar mill effluent versus soil K+
0.73
Sugar mill effluent versus soil Ca2+
0.97
Sugar mill effluent versus soil Mg2+
0.90
Sugar mill effluent versus soil TKN
0.97
Sugar mill effluent versus soil PO43-
0.85
Sugar mill effluent versus soil SO42-
0.95
Sugar mill effluent versus soil Fe2+
0.93
Sugar mill effluent versus soil Cd
0.88
Sugar mill effluent versus soil Cr
0.86
Sugar mill effluent versus soil Cu
0.83
Sugar mill effluent versus soil Mn
0.85
Sugar mill effluent versus soil Zn
0.80
0
2
4
6
8
10
12
14
16
18
20
Cr Zn Cd Mn Cu
Heavy metals
Contamination factor
Figure 1. Contamination factor of heavy metals in soil after fertigation with sugar mill effluent.
Error bars are standard error of the mean.
more organic carbon in the effluent irrigated soil might be
due to the high organic nature of the effluent (Kaushik et
al., 1996; Ayyasamy et al., 2008).
Recent studies by Kaushik et al. (2005) reported that
the distillery effluent irrigation increased the EC, pH, OC,
TKN, P, Na, K, Ca, Mg of the soil. Kannan and Upreti
(2008) found that distillery effluent increased the nutrients
and toxic chemicals in the soil when used for agricultural
irrigation. Effluent irrigation generally adds significant
quantities of salts to the soil environment, such as
sulfates, phosphates, bicarbonates, chlorides of the
cations sodium, calcium, potassium and magnesium;

Kumar and Chopra 096
70
75
80
85
90
95
100
020 40 60 80 100
Distillery effluent concentration (%)
Seed germination (%)
Seed germination
Figure 2. Seed germination of V. faba after fertigation with sugar mill effluent. Error
bars are standard error of the mean.
0
20
40
60
80
100
120
140
160
020 40 60 80 100
Distillery effluent concentration (%)
Relative toxicity (%)
Relative toxicity
Figure 3. Relative toxicity of sugar mill effluent against seed germination of V. faba. Error
bars are standard error of the mean.
they stimulate the growth at lower concentration but
inhibit at higher concentration (Kumar, 2010).
Effect on germination of V. faba
The maximum seed germination (94.00%) of V. faba was
recorded with control (bore well water), while the
minimum seed germination (80.00%) was noted with
100% concentration of sugar mill effluent. The
germination of V. faba was decreased with the increase
in sugar mill effluent concentrations and it was found to
be negatively correlated (r = -0.98) with sugar mill effluent
concentrations. ANOVA analysis showed that increase of
the concentration from 20 to 100% of sugar mill effluent
showed significant (P<0.05) effect on seed germination of
V. faba (Figure 2). Relative toxicity (RT) of sugar mill
effluent was increased when the concentration of sugar
mill effluent increased. The maximum RT (117.50%) was
recorded with 100% concentration of sugar mill effluent
(Figure 3). RT (r = +0.73) of sugar mill effluent was
observed to be positively correlated with different
concentrations of sugar mill effluent.
This type of germination pattern of V. faba is likely due
to the presence of toxicants in the higher concentration of

effluent which may inhibit the germination at higher
concentrations as observed earlier for T. foenum-
graecum crop (Kumar and Chopra, 2012). The findings
are very much in accordance with those of Pandey et al.
(2007) who reported that the germination percentage of
Triticum aestivum, Pisum sativm and Abelmoschus
esculentus decreases with increasing effluent
concentrations. Ramana et al. (2002) observed that
distillery effluent showed inhibitory effect on seed
germination at higher concentrations.
Effect on vegetative growth of V. faba
The maximum shoot length (136.84 cm), root length
(18.26 cm) chlorophyll content (4.88 mg/g.f.wt) and
LAI/plant (5.64) of V. faba were recorded with 40%
concentration of sugar mill effluent at vegetative growth
stage, that is, at 45 days. The minimum shoot length
(105.23 cm), root length (11.42 cm) chlorophyll content
(3.94 mg/g.f.wt) and LAI/plant (4.72) of V. faba were
recorded with control (borewell water). At 100% shoot
length (114.26 cm), root length (13.48 cm) chlorophyll
content (4.26 mg/g.f.wt) and LAI/plant (4.86) of V. faba
were higher than the control but lower than 40%
concentration.
The ANOVA indicated that the concentrations of sugar
mill effluent had significant (P<0.05) effect on shoot
length, root length, chlorophyll content and LAI/plant of V.
faba. Shoot length and chlorophyll content was also
found to be more significantly (P<0.01) affected with
different concentrations of sugar mill effluent. Shoot
length of V. faba was also found significantly (P<0.01)
different with 40 to 100% concentration of sugar mill
effluent. The sugar mill effluent concentration (60 and
80%) also showed significant effect (P<0.01) on LAI of V.
faba.
The maximum growth of V. faba was recorded with
40% concentration of sugar mill effluent and decreased
when the concentration of sugar mill effluent increased. It
may be likely due to the presence of optimum content of
nutrients at 40% concentration of sugar mill effluent.
More irrigation of V. faba plants with higher concentration
of sugar mill effluent decreased the vegetative growth of
V. faba; it is likely due to the presence of high salt content
at these concentrations, which inhibit the growth of V.
faba.
Chlorophyll content was higher due to the use of 40%
sugar mill effluent in both seasons, and is likely due to
Fe, Mg and Mn contents in the sugar mill effluent, which
are associated with chlorophyll synthesis (Porra, 2002).
The findings are supported by Ayyasamy et al. (2008).
Bharagava et al. (2008) also reported that distillery
effluent irrigation increased the chlorophyll and crop yield
in Indian mustard plants (Brassica nigra L.) at the lower
concentrations (25 and 50%) and decreased them at
higher concentrations (75 and 100%) to their respective
controls.
Int. J. Biotechnol. Res. 097
Effect on flowering of V. faba
Among all concentrations of sugar mill effluent, most
flowers (168.00) of V. faba were recorded with 40%
concentration while few flowers (142.00) were noted with
the control. ANOVA analysis showed that sugar mill
effluent concentration significantly (P<0.05) affected the
number of flowers of V. faba. Nitrogen and phosphorus
are essential for flowering. Too much nitrogen can delay,
or prevent, flowering while phosphorus deficiency is
sometimes associated with poor flower production, or
flower abortion. The 40% of sugar mill effluent favored
the flowering of V. faba. This is likely due to this
concentration containing sufficient nitrogen and
phosphorus. Furthermore, nitrogen and phosphorus
prevent flower abortion so pod formation occurs (El-
Naggar, 2005).
Effect on maturity of V. faba
The present study showed that the maximum fresh
yield/plant (150.44 g) and dry yield/plant (34.68 g) of V.
faba was recorded with 40% concentration of sugar mill
effluent while the minimum fresh yield/plant (106.24 g)
and dry yield/plant (18.40 g) of V. faba was recorded with
the control. The 40% concentration of sugar mill effluent
showed significant (P<0.05) effect on fresh yield/plant
and dry yield/plant of V. faba. The maximum fresh
yield/plant and dry yield/plant of V. faba was recorded
with 40% concentration of sugar mill effluent; it is likely
due to the presence of desirable content of nitrogen,
phosphorus and potassium in the sugar mill effluent,
which stimulates the pod formation in V. faba. The
findings are very much in accordance with those of
Ramana et al. (2002).
Translocation of heavy metals in V. faba
ANOVA showed that all concentrations of sugar mill
effluent had significant (P<0.001) effect on the content of
Cd, Cr, Cu, Mn and Zn of V. faba. It is likely due to the
presence of significant quantity of these metals in the
sugar mill effluent and irrigated soil. The content of Cd,
Cr, Cu, Mn and Zn in V. faba was recorded maximum
with 100% sugar mill effluent (Figures 4 to 8). The
translocation of various heavy metals in different parts of
V. faba was in order of leaves>shoot>root>fruits for Cu,
Mn and Zn; root>shoot>leaves>fruit for Cd; and
shoot>root>leaves>fruit for Cr in V. faba after sugar mill
effluent irrigation (Figures 4 to 8). The translocation of Cd
in the root (r = +0.98), shoot (r = +0.99), leaves (r =
+0.94), fruits (r = +0.97), Cr in the root, shoot, fruit (r =
+0.97) and leaves (r = +0.98), Cu in the shoot and leaves
(r = +0.99), root and fruit (r = +0.98), Mn in the root,
shoot, leaves and fruits (r = +0.99) and Zn in the root,
shoot, leaves (r = +0.99) and fruit (r = +0.98), were
recorded to be positively correlated with different

Kumar and Chopra 098
0
2
4
6
8
10
12
020 40 60 80 100
Distillery effluent concentration (%)
Cd content (mg/Kg)
Cd in Shoot Cd in Root
Cd in leaves Cd in fruits
Figure 4. Translocation of Cd in various parts of V. faba after fertigation with sugar
mill effluent. Error bars are standard error of the mean.
0
0.5
1
1.5
2
2.5
3
3.5
4
4.5
020 40 60 80 100
Distillery effluent concentration (%)
Cr content (mg/Kg)
Cr in Shoot Cr in Root
Cr in leaves Cr in fruits
Figure 5. Translocation of Cr in various parts of V. faba after fertigation with
sugar mill effluent. Error bars are standard error of the mean.
0
2
4
6
8
10
12
020 40 60 80 100
Distillery effluent concentration (%)
Cu content (mg/Kg)
Cu in Shoot Cu in Root
Cu in leaves Cu in fruits
Figure 6. Translocation of Cu in various parts of V. faba after fertigation
with sugar mill effluent. Error bars are standard error of the mean.

Int. J. Biotechnol. Res. 099
0
5
10
15
20
25
020 40 60 80 100
Distillery effluent concentration (%)
Mn content (mg/Kg)
Mn in Shoot Mn in Root
Mn in leaves Mn in fruits
Figure 7. Translocation of Mn in various parts of V. faba after fertigation with sugar
mill effluent. Error bars are standard error of the mean.
0
2
4
6
8
10
12
14
16
18
20
020 40 60 80 100
Distillery effluent concentration (%)
Zn content (mg/Kg)
Zn in Shoot Zn in Root
Zn in leaves Zn in fruits
Figure 8. Translocation of Zn in various parts of V. faba after fertigation with sugar mill
effluent. Error bars are standard error of the mean.
concentration of sugar mill effluent. The order of
contamination factor of heavy metals was
Cr>Cu>Cd>Mn>Zn in V. faba after irrigation with sugar
mill effluent concentrations (Figure 9). Chandra et al.
(2009) reported the higher content of Cu, Cd, Cr, Zn, Fe,
Ni and Mn in wheat and mustard plants irrigated with
mixed distillery and tannery effluents. The metals’
contents in various plant parts of V. faba were increased
when the concentration of sugar mill effluent increased.
Effect on biochemical constituents of V. faba
The maximum crude proteins, crude fiber and total
carbohydrates of V. faba were recorded with 40%
concentration of sugar mill effluent (Figure 10). Content
of crude proteins (r = +0.59), crude fiber (r = +0.53) and
total carbohydrates (r = +0.55) of V. faba were noted to
be positively correlated with sugar mill concentration. The
biochemical constituents decreased when the
concentration of sugar mill increased, and it was
recorded maximum with 40% concentration of sugar mill
effluent and it might be due to the optimum uptake of
nutrients at this concentration. The changes in
biochemical constituents were in accordance with
Sukanya and Meli (2004) who reported that the nutrients
quality of wheat decreased when the concentration of

Kumar and Chopra 100
0
2
4
6
8
10
12
14
Cr Cu Cd Mn Zn
Heavy metals
Contamination factor
Shoot Root
Leaves Fruit
Figure 9. Contamination factor of heavy metals in various parts of V. faba after fertigation
with sugar mill effluent. Error bars are standard error of the mean.
0
10
20
30
40
50
60
70
020 40 60 80 100
Distillery effluent concentration (%)
Content (%)
Crude proteins Crude fiber Total carbohydrates
Figure 10. Content of crude proteins, crude fiber and total carbohydrates in V. faba after
fertigation with sugar mill effluent. Error bars are standard error of the mean.
distillery effluent increased.
Conclusions
It was concluded that sugar mill effluent irrigation
increased the EC, pH, OC, HCO3-, CO3-2, Na+, K+, Ca2+,
Mg2+, TKN, PO43-, SO42-, Fe2+, Cd, Cr, Cu, Mn and Zn in
the soil used for the cultivation of V. faba. All effluent
concentrations were better than the control in nutrient
accumulation. The growth performance of V. faba was
gradually increased at lower concentration (that is, from
20 to 40%) and decreased at higher concentrations (that
is, 60 to 80%). It was recorded maximum with 40%
concentration of sugar mill effluent. Thus, there is certain
growth stimulating, as well as inhibiting, substances
present in the sugar mill effluent which are responsible
for this growth pattern. The biochemical components
namely: crude proteins, crude fiber and total
carbohydrates were noted maximum with 40%
concentration of sugar mill effluent. The contamination

factor of heavy metals was in order of Cr>Zn>Cd>Mn>Cu
for soil and Cr>Cu>Cd>Mn>Zn for V. faba after fertigation
with sugar mill effluent. The translocation of various
heavy metals in different parts of V. faba were in order of
leaves>shoot>root>fruits for Cu, Mn and Zn;
root>shoot>leaves>fruit for Cd; and
shoot>root>leaves>fruit for Cr after sugar mill effluent
irrigation. Thus, the sugar mill effluent can be used with
appropriate dilution for the maximum yield of V. faba.
Therefore sugar mill effluent can be used for irrigation
purposes after proper dilution for the maximum yield of V.
faba. Further investigation is required on the agronomical
practices and biochemical changes in V. faba after
irrigation with sugar mill effluent.
ACKNOWLEDGEMENT
The University Grants Commission, New Delhi, India is
acknowledged for providing the financial support in the
form of UGC research fellowship (F.7-70/2007-2009
BSR) to the corresponding author.
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