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Communications in Soil Science and
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Ferti-irrigation Effect of Paper Mill
Effluent on Agronomical Practices of
Phaseolus vulgaris (L.) in Two Seasons
Vinod Kumara & A. K. Chopraa
a Agro-ecology and Pollution Research Laboratory, Department
of Zoology and Environmental Science, Faculty of Life Sciences,
Gurukula Kangri University, Haridwar (Uttarakhand), India
Accepted author version posted online: 13 Jun 2014.Published
online: 21 Aug 2014.
To cite this article: Vinod Kumar & A. K. Chopra (2014) Ferti-irrigation Effect of Paper Mill Effluent on
Agronomical Practices of Phaseolus vulgaris (L.) in Two Seasons, Communications in Soil Science and
Plant Analysis, 45:16, 2151-2170, DOI: 10.1080/00103624.2014.929698
To link to this article: http://dx.doi.org/10.1080/00103624.2014.929698
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Communications in Soil Science and Plant Analysis, 45:2151–2170, 2014
Copyright © Taylor & Francis Group, LLC
ISSN: 0010-3624 print / 1532-2416 online
DOI: 10.1080/00103624.2014.929698
Ferti-irrigation Effect of Paper Mill Effluent
on Agronomical Practices of Phaseolus vulgaris (L.)
in Two Seasons
VINOD KUMAR AND A. K. CHOPRA
Agro-ecology and Pollution Research Laboratory, Department of Zoology and
Environmental Science, Faculty of Life Sciences, Gurukula Kangri University,
Haridwar (Uttarakhand), India
The potential of agrobased paper mill effluent (PME) as ferti-irrigant was assessed.
Ferti-irrigation responses to 5, 10, 25, 50, 75, and 100% of PME doses on Phaseolus
vulgaris L., cv. Annapurna, in the rainy and summer seasons were investigated. The
fertigant concentrations produced changes in electrical conductivity (EC), pH, organic
carbon (OC), sodium (Na+), potassium (K+), calcium (Ca2+), magnesium (Mg2+), total
Kjeldahl nitrogen (TKN), phosphate (PO43–), sulfate (SO42–), iron (Fe2+), cadmium
(Cd), chromium (Cr), copper (Cu), manganese (Mn), and zinc (Zn) of the soil in both
seasons. The agronomic performances of P. vulgaris increased from 5 to 25% in both
seasons compared to controls. The accumulation of metals increased in soil and P. vul-
garis from 5 to 100% PME concentrations in both seasons. The contamination factor
(Cf) of various metals was in order of Cr >Mn >Cu >Cd >Zn for soil and Mn >Zn
>Cu >Cd >Cr for P. vulgaris in both seasons after fertigation with PME. Therefore,
PME can be used to improve the soil fertility and yield of P. vulgaris after appropriate
dilution.
Keywords Ferti-irrigation, paper mill effluent, Phaseolus vulgaris, metals, rainy
season, summer season
Introduction
The pulp and paper industry is considered one of the most polluting industries all over
the world (Kamalakar, Sharma, and Melkania 1991; Chaudhary et al. 2002; Kumar and
Chopra 2012). During the chemical pulping and paper-making process, a huge quantity
of effluent is released from these mills (Singh, Marwaha, and Khanna 1996; Dutta and
Biossya 1999; Pokhrel and Viraraghavan 2004). At present, there are 666 pulp and paper
mills in India, of which 632 units are agro-residue- and recycled-fiber-based units. They
generate a huge amount of wastewater (black liquor) having high chemical oxygen demand
(COD) and biochemical oxygen demand (BOD) values (Elisa, Vanderlei, and Nelson 1991;
Fazeli et al. 1998). In India agro-based pulp and paper mills are one of the most pollut-
ing industries; in addition, they are high consumers of raw water and discharge a huge
Received 21 February 2013; accepted 20 March 2014.
Address correspondence to Vinod Kumar, Agro-ecology and Pollution Research Laboratory,
Department of Zoology and Environmental Science, Faculty of Life Sciences, Gurukula Kangri
University, Haridwar 249404 (Uttarakhand), India. E-mail: drvksorwal@gmail.com
Color versions of one or more of the figures in the article can be found online at www.
tandfonline.com/lcss
2151
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2152 V. Kumar and A. K. Chopra
amount of chlorinated lignosulfonic acids, chlorinated resin acids, chlorinated phenols,
and chlorinated hydrocarbon in the effluent (Srivastava 1991; Singh et al. 2002).
Applications of industrial wastes as fertilizer and soil amendment have become pop-
ular in agriculture (Baruah and Das 1997; Dutta and Biossya 1998). Effluents from these
industries contain appreciable amounts of metals such as copper, cadmium, iron, lead,
manganese, nickel, and zinc (Sauve, Henderson, and Allen 2000). Irrigation with such
effluents increases organic carbon content and metal accumulation in soil and agricultural
crops (Hati et al. 2007; Chopra, Pathak, and Parasad 2009). Application of wastewater
in agricultural fields may be a viable method of disposal and would sustain agriculture
in nonirrigated areas, where availability of fresh water is scarce. In agriculture, irrigation
water can affect soil characteristics and agricultural crop growth (Ramana et al. 2002).
Water effects on soils and crops are of more concern to people when the irrigant is wastew-
ater, which may contain elements capable of inducing adverse effects on the soil and the
agricultural products. The various elements introduced via paper mill wastewater irrigation
affect not only the crop growth and soil properties but also their relative mobility in the soil
profile (Kumar and Chopra 2010). Long-term irrigation with effluents increases accumu-
lation of metals in soil and increases chances of their entrance in food chain (Chopra,
Pathak, and Parasad 2009). Amounts of metals mobilized in the soil environment are
functions of pH, clay content, organic matter, cation exchange capacity, and other soil
properties, making each soil unique in terms of pollution management (Howe and Wagner
1996).
The metals copper (Cu), iron (Fe), nickel (Ni), zinc (Zn), and other trace elements are
essential for proper functioning of biological systems, and their deficiency, or excess, could
lead to a number of disorders (Kimberly and William 1999). Metals are capable of forming
insoluble complex compounds with soil organic matter, and contents of cadmium (Cd),
Cu, Ni, manganese (Mn), and Zn are dependent on pH of soil solution and soil organic
matter (Gomathi and Oblisami 1992; Sauve, Henderson, and Allen 2000). Soil type is
one of the most important factors to determine metal content of food plants (Itanna 2002;
Madyiwa et al. 2002). However, metals content in plants can be affected by other factors
such as the application of fertilizers, sewage sludge, or irrigation with wastewater (Devkota
and Schmidt 2000; Frost and Ketchum 2000). Contamination of agricultural soils with
metals can pose long-term environmental problems and is not without health implications
(Ferguson 1990; Chopra, Pathak, and Parasad 2009).
French bean (Phaseolus vulgaris L.) is cultivated during the rainy and summer seasons
in India, one sown at the end of February for the summer crop and at the end of August
for the rainy season crop. During the past decade, cultivation of P. vulgaris has increased
due to its short growing period, biannual growing habit, and high acceptance (Carai et al.
2009).
Some crops have greater potential yields with wastewater irrigation, which reduces the
need for chemical fertilizers and results in net cost savings to farmers (Kannan and Upreti
2008). It is important to understand the specificity of crop–effluent interaction for appro-
priate applications in irrigation (Kumar and Chopra 2012; Ramana et al. 2002). In some
studies, characteristics of effluent of industries and agronomic properties of various crop
plants have been determined (Sundri and Kanakarani 2001;Hatietal.2007; Kannan and
Upreti 2008). Most studies were conducted on few agronomic stages with limited param-
eters in various crops, but there are few reports on comprehensive agronomic studies at
various agronomic stages of these plants (Kaushik et al. 2005). Use of industrial effluents
on cultivation of P
. vulgaris is receiving attention (Nwoko et al. 2007) but additional infor-
mation is needed on how this crop responds to various concentrations of different types
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Ferti-irrigation Effect of Paper Mill Effluent 2153
of effluents. The investigation was undertaken to study ferti-irrigation effect of paper mill
effluent on agronomical practices of Phaseolus vulgaris (L.) in two seasons.
Materials and Methods
Experimental Design
The study was conducted at the Experimental Garden of the Department of Zoology and
Environmental Sciences, Faculty of Life Sciences, Gurukula Kangri University Haridwar,
India (29◦5510.81 N and 78◦0708.12 E), to determination effects of fertigation with
paper mill effluent (PME) on P. vulgaris. The crop was cultivated in the summer and rainy
seasons in 2008 and 2009. Poly bags (dia. 30 cm) were used. The experiment was replicated
six times in each season. A distance of 30 cm between bags and 60 cm between treatments
was maintained. Holes were punched in bags for aeration and drainage.
Effluent Collection and Analysis
The effluent samples were provided from the Star Paper Mill, Saharanpur (Uttar Pradesh),
which produces paper from agricultural waste or residues. Effluent waste was collected
from a settling tank installed on the campus, by the paper mill, to reduce biological oxy-
gen demand (BOD) and solids from the paper mill in plastic container, and was brought
to the laboratory and analyzed for total dissolved solids (TDS), pH, electrical conductiv-
ity (EC), dissolved oxygen (DO), BOD, COD, chlorides (Cl−), bicarbonates (HCO3−),
carbonates (CO32–), sodium (Na+), potassium (K+), calcium (Ca2+), magnesium (Mg2+),
total Kjeldahl nitrogen (TKN), nitrate (NO32–), phosphate (PO43– ), sulfate (SO42–), cad-
mium (Cd), chromium (Cr), copper (Cu), iron (Fe), manganese (Mn), zinc (Zn), standard
plate count (SPC), and most probable number (MPN) following standard methods (APHA
2005; Chaturvedi and Sankar 2006) before use as fertigant.
Soil Preparation, Filling of Bags, Sampling, and Analysis
The loamy soil was collected from a depth of 0–15 cm. Each bag (30 ×30 cm) was filled
with 5 kg of soil, which had been air dried and sieved to remove debris, and mixed with
equal quantity of cow manure. The soils in each bag were fertigated twice a week with
500 mL of PME with 5, 10, 25, 50, 75, and 100%, along with well water as the con-
trol. The soil was analyzed prior to planting and after harvest for various physicochemical
parameters such as soil texture, bulk density (BD), water-holding capacity (WHC), EC, pH,
OC, Na+,K
+,Ca
2+,Fe
2+,Mg
2+,PO
43–,SO
42–, TKN, Cd, Cr, Cu, Mn, and Zn determined
following standard methods cited in Chaturvedi and Sankar (2006).
Sowing of Seeds, Irrigation Pattern, and Collection of Crop Parameter Data
Seed of P. vulgaris were sown at the end of February 2008 and 2009 for the summer crop
and at the end of August 2008 and 2009 for the rainy season crop. Seed of P. vulgaris,
cv. Annapurna, were procured from Indian Council of Agriculture Research (ICAR), Pusa,
New Delhi; sterilized with 0.01% mercuric chloride; and soaked in water for 12 h. Seven
seeds were sown in each poly bag with 7.5 cm between plants, thinned to five plants, and
replicated six times. The agronomic parameters at different stages (0–90 days) were deter-
mined following standard methods for seed germination, shoot length, root length, number
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2154 V. Kumar and A. K. Chopra
of flowers, number of pods, and crop yield (Chandrasekar, Subramani, and Saravana 1998);
dry weight (Milner and Hughes 1968); chlorophyll content (Porra 2002); relative toxicity
(RT) (Chapagain 1991); leaf area index (LAI) (Denison and Russotti 1997); and harvest
index (HI) (Sinclair 1998). The nutrient quality of crop was determined by using the fol-
lowing parameters: crude protein (4.204 Anonymous 1980), crude fiber (4.601 Anonymous
1980) and the total carbohydrates in dry matter were determined by the anthrone reagent
method (Cerning and Guilhot 1973).
Extraction of Metals and their Analysis
For metal analysis, 5–10 mL PME and 0.5–1.0 g of air-dried soil or plants were digested in
tubes with 3 mL of concentrate nitric acid (HNO3) digested in an electrically heated block
for 1 hr at 145 ◦C. To this mixture 4 mL of perchloric acid (HClO4) was added and heated to
240 ◦C for 1 h. The mix was cooled and filtered through Whatman No. 42 filter paper (GE
Healthcare, Little Chalfont, UK), made up to 50 mL, and used for analysis. Metals were
analyzed using an atomic absorption spectrophotometer (Perkin-Elmer, Analyst 800 AAS,
GenTech Scientific Inc., Arcade, N.Y.) following methods of APHA (2005) and Chaturvedi
and Sankar (2006). The contamination factor (Cf) for metals accumulated in PME irrigated
soil and P. vulgaris was calculated following Håkanson (1980).
Data Analysis
Data were analyzed with SPSS (ver. 12.0, SPSS Inc., Chicago, Ill., USA). Data were sub-
jected to one-way analysis of variance (ANOVA). Mean standard deviation and coefficient
of correlation (r-value) of soil and crop parameters with effluent concentrations were cal-
culated with MS Excel (ver. 2003, Microsoft Redmond Campus, Redmond, Wash.) and
graphs were produced with Sigma Plot (ver. 12.3, Systat Software, Inc., Chicago, Ill.).
Results and Discussion
Characteristics of Effluent
Values of physicochemical and microbiological parameters varied over PME concentra-
tion (Table 1). The PME effluent was alkaline (pH 8.48). The alkaline nature of the PME
might be due to presence of high concentrations of organic acids. The BOD, COD, Cl−,
Ca2+,Fe
2+, TKN, SO42–, MPN, and SPC were above the prescribed limits of the Indian
Irrigation Standards (BIS 1991). High BOD and COD might be due to presence of high
oxidizable organic matter and rapid consumption of dissolved inorganic materials. The
greater bacterial load (SPC and MPN) in PME might be due to presence of more dissolved
solids and organic matter in effluent as earlier reported by Kumar and Chopra (2010). The
TKN, PO43–,K
+,Ca
2+, and Mg2+in effluent were greater than the prescribed standards
(Table 1). In the present study, the contents of BOD, COD, TKN, Cl−,SO
42–, and PO43–
were more in PME then the content of BOD (29.00 mgL−1), COD (248.00 mgL−1), total
nitrogen (12.04 mgL−1), chlorides (100.02 mgL−1), sulfate (12.00 mgL−1), and phosphate
(0.75 mgL−1) in paper mill effluent reported by Patterson et al. (2008). In the case of met-
als, the contents of Cd, Cr, Cu, Fe2+, Mn, and Zn were greater than permissible limits for
industrial effluent (BIS 1991). The contents of these metals in paper mill effluent were also
greater then the contents of Zn (14.18 mgL−1), Cd (2.26 mgL−1) and Cu (7.17 mgL−1)in
paper mill effluent reported by Howe and Wagner (1999).
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Table 1
Physicochemical and microbiological characteristics of paper mill effluent (PME)
Effluent concentration (%)
Parameter 0 (BWW)a5 10255075100
BISbfor
irrigation water
TDS (mg L−1) 221.5 1362.00 1786.00 2245.00 2511.00 2879.00 3984.00 1900
EC (dS m−1) 0.3 2.12 2.72 3.47 3.69 4.84 6.12 —
pH 7.5 7.54 7.65 7.90 7.95 8.12 8.48 5.5−9.0
DO (mg L−1) 8.24 5.36 4.22 3.53 2.36 1.68 NIL —
BOD (mg L−13.8 70.39 129.00 307.50 618.00 923.50 1226.50 100
COD (mg L−1) 5.9 145.00 283.00 710.00 1421.00 2123.00 2832.50 250
Cl −(mg L−1) 15.7 64.50 104.25 213.75 322.00 632.50 839.50 500
HCO3−(mg L−1) 282.0 264.96 275.67 305.93 371.40 415.74 546.00 —
CO32– (mg L−1) 105.8 119.70 130.41 159.99 173.43 180.30 194.43 —
Na+(mg L−1) 9.7 34.87 62.51 128.25 246.12 367.60 4815 —
K+(mg L−1) 5.5 19.42 23.06 41.29 77.99 99.54 126.75 —
Ca2+(mg L−1) 23.5 60.47 70.68 140.70 249.99 344.87 439.50 200
Mg2+(mg L−1) 12.2 19.24 22.08 34.10 50.38 62.19 74.22 —
TKN (mg L−1) 24.3 33.02 37.46 51.12 69.62 81.19 92.54 100
NO32– (mg L−1) 25.2 50.03 64.43 110.91 218.29 288.46 378.50 100
PO43– (mg L−1) 0.04 8.75 18.37 41.79 80.81 123.87 160.25 —
SO42– (mg L−1) 17.6 51.31 79.12 176.05 323.14 478.70 633.50 1000
Fe2+(mg L−1) 0.3 0.70 1.64 3.98 7.83 11.72 15.25 1.0
Cd (mg L−1) 0.06 0.32 0.64 1.54 3.09 4.65 6.42 15
Cr (mg L−1) 0.1 1.30 1.98 1.77 2.14 2.75 2.98 2.00
Cu (mg L−1) 0.04 1.04 1.59 1.98 2.34 2.98 3.57 3.00
Mn (mg L−1) 0.02 0.88 1.29 1.82 2.51 2.96 3.24 1.00
Zn (mg L−1) 0.04 0.12 0.83 1.30 1.96 2.29 2.78 2.00
SPC (SPC mL−1)633.8×1045.3 ×1057.4 ×1064.6 ×1072.4 ×1083.6 ×1010 10000
MPN (MPN 100 mL−1)2.6×1014.9 ×1036.8 ×1038.4 ×1034.6 ×1046.6 ×1054.6 ×1065000
aBWW, bore well water.
bBIS, Bureau of Indian Standards. Least squares means analysis.
2155
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2156 V. Kumar and A. K. Chopra
Characteristics of Soil
Physicochemical characteristics of the soil changed due to irrigation with PME
(Tables 2–7). At harvest (90 days after sowing) there was no significant change in the soil
texture (loamy; 40% sand, 40% silt, 20% clay). Irrigation with 100% PME had the most
reduction in WHC and BD and increase in EC, OC, Na+,K
+,Ca
2+,Mg
2+,Fe
2+, TKN,
PO43–,SO
42–, Cd, Cr, Cu, Mn, and Zn in both seasons (Tables 5 and 6). Values of WHC
and BD were insignificantly changed by the different concentrations of PME in both the
cultivated seasons. Values of WHC and BD were reduced from their initial (control) values
45.58% and 1.42 gm cm−3to 39.88, 38.80%, and 1.40 gm cm−3, respectively, with 100%
PME concentration. Seasons affected OC and TKN; interaction of PME concentration and
the seasons affected EC, pH, OC, and TKN but not WHC and BD (Table 2).
Water-holding capacity (WHC) is related to the number and size distribution of soil
pores, soil moisture content, textural class, structure, salt content, and organic matter. The
BD of soil changes with application of organic manure to soil that substantially modifies
and lowers the soil bulk density. It is used for determining the amount of pore space and
water storage capacity of the soil. Organic matter supplied through the distillery effluent
and other kinds of wastes, such as sludge, can lower the BD and WHC (Kumar and Chopra
2012).
Table 2
ANOVA for effect of PME on soil characteristics
Source WHC BD EC pH OC TKN
Season (S) ns ns ns ns ∗∗
PME concentration (C) ns ns ∗∗ ∗ ∗∗ ∗∗
Interaction S ×Cnsns
∗ ∗ ∗∗ ∗∗
Notes. ns, ∗,and∗∗ denote nonsignificant or significant at P≤0.05 or P≤0.01 by ANOVA,
respectively.
Table 3
ANOVA for effect of PME on concentrations of cations and anions
Source Na+K+Ca2+Mg2+Fe2+PO43– SO42–
Season (S) ∗∗ ∗ ∗ ∗ ∗ ∗
PME concentration (C) ∗∗ ∗ ∗ ∗ ∗∗ ∗∗ ∗∗
Interaction S ×C∗∗ ∗∗ ∗∗ ∗∗ ∗∗ ∗∗ ∗∗
Notes. ∗and ∗∗ denote significant at P≤0.05 or P≤0.01 by ANOVA, respectively.
Table 4
ANOVA for effect of PME on concentrations of metals
Source Cd Cr Cu Mn Zn
Season (S) ∗∗∗ns ∗
PME concentration (C) ∗∗ ∗∗ ∗∗ ∗ ∗∗
Interaction S ×C∗∗ ∗∗ ∗∗ ∗∗ ∗∗
Notes. ∗and ∗∗ denote significant at P≤0.05 or P≤0.01 by ANOVA, respectively.
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Ferti-irrigation Effect of Paper Mill Effluent 2157
Table 5
Effects of PME concentration and season interaction on physicochemical characteristics
of a loamy soil before and after irrigation of P. vulgaris in both seasons
Season ×
%PME
EC
(dS m−1)pH
OC
(mg kg−1)
Na+
(mg kg−1)
K+
(mg kg−1)
Ca2+
(mg kg−1)
Mg2+
(mg kg−1)
Rainy
0 2.06 7.50 0.43 17.56 154.09 14.11 1.68
5 2.42ns 8.09ns 1.53∗22.22ns 162.04ns 34.59ns 3.80ns
10 2.60ns 8.23ns 2.95∗24.34∗171.09ns 37.55ns 6.98ns
25 2.72∗8.33ns 4.17∗∗ 26.50∗184.61∗73.10∗9.83∗
50 2.95∗8.44∗5.11∗∗ 29.08∗209.95∗103.73∗12.14∗
75 3.09∗8.68∗7.61∗∗ 30.62∗∗ 217.78∗∗ 132.08∗15.38∗
100 3.17∗∗ 8.73∗8.93∗∗ 34.91∗∗ 225.97∗∗ 148.99∗∗ 22.20∗
Summer
0 2.09 7.52 0.45 17.68 155.29 14.17 1.69
5 2.48ns 8.10ns 1.66∗24.92ns 164.54ns 37.09ns 4.05ns
10 2.67ns 8.37ns 3.18∗26.84∗175.59ns 45.48ns 7.23ns
25 2.79∗8.55ns 4.35∗∗ 29.00∗189.11∗77.60∗11.08∗
50 3.07∗8.68∗5.29∗∗ 31.58∗218.45∗109.23∗14.39∗
75 3.17∗8.72∗7.88∗∗ 33.12∗∗ 227.28∗∗ 138.58∗17.63∗
100 3.24∗∗ 8.84∗9.15∗∗ 37.41∗∗ 236.47∗∗ 156.99∗∗ 24.45∗
Notes. ns, ∗,and∗∗ denote nonsignificant or significant at P≤0.05 or P≤0.01 by ANOVA,
respectively. Least squares means analysis.
Fertigation with 100% effluent concentration decreased BD (1.40%), pH (16.66–
17.28%), and WHC (13.26–15.61%) and increased EC (53.88–55.02%), OC (1933.33–
1976.74%), Na+(98.80–111.59%), K+(46.64–52.27%), Ca2+(955.91–1007.90%), Mg2+
(1221.42–1346.74%), TKN (818.79–832.07%), PO43– (146.55–155.61%), SO42– (70.02–
84.65%), Fe2+(118.25–127.75%), Cd (287.32–344.59%), Cr (695.23–761.36%), Cu
(326.60–378.36%), Mn (534.61–607.40%), and Zn (118.26–180.95%) in the soil in both
seasons.
The 10 to 100% PME concentrations affected EC, OC, TKN, K+,Ca
2+,PO
43–,SO
42,
Zn, and Cd in P. vulgaris cultivated soil in both seasons. The 5% PME concentration
affected EC and TKN in both seasons. The 25 to 100% PME concentrations affected Na+,
Mg2+,Fe
2+, Cu, and Cr contents in the soil in both seasons. Soil pH was affected by the
50, 75, and 100% PME concentrations whereas Mn was affected by the 75 and 100% PME
concentrations. The soil Na+was affected by the 5% PME in summer season (Tables 5 and
6). The EC, OC, Na+,K
+,Ca2+,Mg
2+,Fe
2+, TKN, PO43–,SO
42– Zn, Cd, Cu, Mn, and
Cr positively correlated with PME concentration in both seasons (Table 7). In the present
study, more irrigation of P. vulgaris considerably increased the contents of OC, Na+,K
+,
Ca2+,Mg
2+,Fe
2+, TKN, PO43–,SO
42– Zn, Cd, Cu, Mn, and Cr in soil. Season, PME con-
centration, and the their interaction affected the cations Na+,K
+,Ca
2+,Mg
2+, and Fe2+
and anions PO43– and SO42– of the soil (Tables 2 and 3).
Kaushik et al. (2005) reported that distillery effluent increased EC, pH, total organic
carbon (TOC), total Kjeldahl nitrogen (TKN), and available phosphorus (P), exchangeable
Na, K, Ca, and Mg in soil. Effluent irrigation generally adds PO43–,HCO
3−,Cl
−,Na+,
Ca2+,K
+,Mg
2+, Zn, Cd, Cr, Cu, Ni, and Mn to the soil (Patterson et al. 2008; Chopra,
Pathak, and Parasad 2009).
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Table 6
Effects of PME concentration and season interaction on physicochemical characteristics of a loamy soil before and after irrigation of
P. vulgaris in both seasons
TKN PO43– SO42– Fe2+Cd Cr Cu Mn Zn
Season ×%PME (mg kg−1)(mgkg
−1)(mgkg
−1)(mgkg
−1)(mgkg
−1)(mgkg
−1)(mgkg
−1)(mgkg
−1)(mgkg
−1)
Rainy
0 30.96 51.75 73.12 2.63 0.71 0.84 2.03 0.26 1.04
5 53.33ns 64.90ns 75.86ns 3.21∗1.34∗1.57∗3.25∗1.08∗1.27∗
10 61.69∗∗ 71.37ns 78.84ns 3.44∗2.06∗2.07∗3.46∗1.15∗1.39∗
25 121.34∗∗ 92.94∗83.62∗5.08∗∗ 2.17∗∗ 3.39∗∗ 5.67∗∗ 1.31∗∗ 1.52∗∗
50 177.27∗∗ 105.72∗∗ 94.86∗5.26∗∗ 2.23∗∗ 4.48∗∗ 5.96∗∗ 1.42∗∗ 1.67∗∗
75 244.32∗∗ 115.11∗∗ 111.38∗∗ 5.58∗∗ 2.42∗∗ 5.68∗∗ 6.22∗∗ 1.55∗∗ 2.15∗∗
100 284.46∗∗ 127.59∗∗ 124.32∗∗ 5.74∗∗ 2.75∗∗ 6.78∗∗ 8.66∗∗ 1.65∗∗ 2.27∗∗
Summer
0 31.86 52.85 74.62 2.63 0.74 0.88 2.08 0.27 1.05
5 55.83ns 67.40ns 78.36ns 3.46∗2.02∗1.78∗3.68∗1.19∗1.36∗
10 64.19∗∗ 78.87∗86.34∗3.69∗2.11∗2.56∗3.81∗1.45∗1.49∗
25 127.84∗∗ 99.44 89.12∗5.33∗∗ 2.19∗∗ 4.23∗∗ 5.95∗∗ 1.56∗∗ 1.62∗∗
50 184.77∗∗ 112.22∗∗ 108.36 5.51∗∗ 2.36∗∗ 5.64∗∗ 6.18∗∗ 1.78∗∗ 1.79∗∗
75 256.82∗∗ 127.61∗∗ 123.87∗∗ 5.83∗∗ 2.97∗∗ 6.75∗∗ 7.42∗∗ 1.87∗∗ 2.33∗∗
100 296.96∗∗ 135.09∗∗ 137.79∗∗ 5.99∗∗ 3.29∗∗ 7.58∗∗ 9.95∗∗ 1.91∗∗ 2.95∗∗
Notes. ns, ∗,and∗∗ denote nonsignificant or significant at P≤0.05 or P≤0.01 by ANOVA, respectively. Least squares means analysis.
2158
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Ferti-irrigation Effect of Paper Mill Effluent 2159
Table 7
Coefficient of correlation (r) between PME and soil characteristics in both seasons
PME soil characteristics Season r value
PME versus soil WHC Rainy −0.94
Summer −0.95
PME versus soil BD Rainy −0.95
Summer −0.96
PME versus soil EC Rainy +0.91
Summer +0.89
PME versus soil pH Rainy +0.89
Summer +0.89
PME versus soil OC Rainy +0.89
Summer +0.89
PME versus soil Na+Rainy +0.95
Summer +0.93
PME versus soil K+Rainy +0.96
Summer +0.96
PME versus soil Ca2+Rainy +0.98
Summer +0.98
PME versus soil Mg2+Rainy +0.97
Summer +0.98
PME versus soil TKN Rainy +0.99
Summer +0.99
PME versus soil PO43– Rainy +0.95
Summer +0.95
PME versus soil SO42– Rainy +0.99
Summer +0.99
PME versus soil Fe2+Rainy +0.88
Summer +0.87
PME versus soil Cd Rainy +0.87
Summer +0.87
PME versus soil Cr Rainy +0.87
Summer +0.87
PME versus soil Cu Rainy +0.96
Summer +0.97
PME versus soil Mn Rainy +0.97
Summer +0.97
PME versus soil Zn Rainy +0.97
Summer +0.97
The soil pH is an important parameter as many nutrients are available to plants only
within a particular pH range. A pH range of 6.0 to 8.3 enhances nutrient availability for
plants, and a pH below 6.0 and above 8.3 inhibits the availability of nutrients for plants
(Charman and Murphy 1991). In the present study pH of the soil was between 6.22 and
6.25 after irrigation with 100% PME, which may increase various soil nutrients.
Total average organic-matter content in the soil irrigated with effluent was greater than
the soil irrigated with bore well water. The more organic matter in effluent-irrigated soil
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2160 V. Kumar and A. K. Chopra
0
2
4
6
8
10
12
Cd Cr Cu Mn Zn
Metals
Contamination factor (Cf)
Cf in rainy season
Cf in summer season
Figure 1. Contamination factor of the metals in soil after irrigation with PME. Error bars are
standard error of the mean.
might be due to the high organic nature of the effluent. Kumar and Chopra (2012) found
the organic content in the soil irrigated with distillery effluent to be greater than in the soil
irrigated with bore well water. Average values of TKN, PO43–, and K+in the soil irrigated
with effluent were found to be greater than in soil irrigated with bore well water. The high
amount of TKN, PO43–, and K+in the soil was due to irrigation with distillery effluents rich
in TKN, PO43–, and K+. The contents of Na+and SO42– were greater in the soil irrigated
with PME, indicating a link between soil Na+and SO42– and greater EC in the PME.
The Cd, Cr, Cu, Mn, and Zn contents in the soil increased as effluent concentration
increased (Table 6). Season, PME concentration, and their interaction affected the Cd, Cr,
Cu, Mn, and Zn in soil (Table 4). The concentration of Cu was maximum whereas that of
Mn was lowest after PME irrigation in both seasons. The contamination factor (Cf) of the
metals indicated that Cr was greatest whereas Zn was lowest in both seasons after irrigation
with 100% PME. The Cf of metals were in the order of Cr >Mn >Cu >Cd >Zn after
irrigation with PME in both seasons (Figure 1). The concentrations of metals Fe, Zn, Mn,
Cu, Pb and Cd were greater in soil irrigated with effluent than in soil irrigated with control
water. Thus, fertigation with distillery effluent increased nutrients as well as metals content
in soil.
Effect on Germination
At 0–15 days after sowing, the best germination (96 and 94%) was with control (BWW)
and the least (82 and 78%) was due to treatment with 100% PME (Figure 2). Germination
was negatively correlated (r =–0.93) with PME concentrations in both seasons. The anal-
ysis of variance (ANOVA) indicated that season had no effect on seed germination and
relative toxicity. The PME concentration and their interaction with season affected seed
germination of P. vulgaris but not relative toxicity (Table 8).
The maximum RT (117.07 and 120.51%) of PME against germination was for the
100% PME (Figure 3) and it was positively correlated (r =+0.52 and r =+0.54) with
PME concentrations in both seasons. The findings are very much in accordance with Malla
and Mohanty (2005), who reported that the germination of green gram (Phaseolus anreus
L.) was decreased as concentration of the paper mill effluent increased from 0 to 100%.
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Ferti-irrigation Effect of Paper Mill Effluent 2161
0
20
40
60
80
100
120
140
05 10255075100
PME concentrations (%)
Seed germination (%)
Germination in rainy season
Germination in summer season
Figure 2. Seed germination of P. vulgaris after irrigation with PME. Error bars are standard error of
the mean.
Table 8
ANOVA for effect of PME on germination and vegetative growth of P. vulgaris
Seed
germination
Relative
toxicity
Shoot
length
Root
length
Dry
weight
Chlorophyll
content LAI
Season (S) ns ns ns ns ns ns ns
PME concentration
(C)
∗ns ∗ns ns ns ∗
Interaction
S×C∗ns ∗ns ns ns ∗
Notes. ns and ∗denote nonsignificant or significant at P≤0.05 by ANOVA, respectively.
0
20
40
60
80
100
120
140
160
0 5 10 25 50 75 100
PME concentration (%)
Relative toxicity (%)
Relative toxicity in rainy season
Relative toxicity in summer season
Figure 3. Relative toxicity of PME against seed germination of P. vulgaris. Error bars are standard
error of the mean.
In the present investigation, the greater concentration of PME did not support seed
germination. The greater concentration of PME lowered emergence of P. vulgaris likely
due to presence of high salt content in the effluent at these concentrations. Seeds take
up water during germination and hydrolyze stored food material to activate enzymatic
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2162 V. Kumar and A. K. Chopra
systems. During germination, salts can inhibit germination. The mechanism of inhibition
of germination by sodium chloride (NaCl) may be related to radicle emergence due to
insufficient water absorption or to toxic effects on the embryo. Seeds that absorb an insuf-
ficient amount of water can accumulate a large amount of Cl−when the osmotic pressure
of the substrate is increased by salt concentration. As a result, the seeds emerge slowly and
at greater concentrations do not germinate (Patterson et al. 2008). High concentrations are
usually most damaging to young plants but not necessarily at germination, although high
salt concentration can slow germination by several days or completely inhibit it. Because
soluble salts move readily with water, evaporation moves salts to the soil surface where
they accumulate and harden the soil surface, which delays germination (Thompson et al.
2001; Kaushik et al. 2005).
Effect on Vegetative Growth
Vegetative growth at 45 days was affected in both seasons (Table 8). Average least shoot
length (20.45 and 23.69 cm), root length (10.48 and 12.20 cm), dry weight (6.23 and
7.46 g), chlorophyll content (2.78 and 3.05 mg g−1fresh weight; f.wt), and LAI plant−1
(2.28 and 2.36) of P. vulgaris were with control while moderate shoot length (32.15 and
34.86 cm), root length (12.47 and 12.75 cm), dry weight (7.68 and 9.86 g), chlorophyll con-
tent (3.84 and 3.92 mg g−1fresh weight), and LAI plant−1(3.74 and 3.82) of P. vulgaris
were with 100% PME in both seasons.
Maximum shoot length (36.58 and 38.95 cm), root length (16.42 and 17.86 cm), dry
weight (11.47 and 13.56 g), chlorophyll content (4.67 and 5.24 mg g−1f.wt), and LAI
plant−1(4.96 and 5.12) of P. vulgaris were due to treatment with the 25% concentration
of PME in both seasons. For all the parameters, the lower value is for the rainy season and
the higher value is for the summer season. The ANOVA indicated that PME concentration
affected shoot length and LAI/plant of P. vulgaris (Table 8). Season had no effect on
shoot length, root length, dry weight, chlorophyll content, and LAI of P. vulgaris.The
interaction of season and PME concentrations only affected shoot length and LAI of P.
vulgaris (Table 8).
Shoot length, dry weight, chlorophyll content, and LAI plant−1of P. vulgaris were
positively correlated with PME concentrations in both seasons (Table 9). Root length was
positively correlated with PME concentrations in rainy season but negatively correlated in
summer season (Table 9). Nwoko et al. (2007) reported the maximum chlorophyll content
in P. vulgaris at 10% concentration of spent engine oil. Malla and Mohanty (2005) reported
that paper mill effluent irrigation increases chlorophyll and protein contents in Indian mus-
tard plants (Phaseolus anreus L.) at the 25 and 50% PME concentrations followed by a
decrease at 75 and 100% PME. The findings were also supported by Reddy and Borse
(2001), who reported that the growth of Trigonella foenum-gracecum (L.) decreased when
concentration of paper mill increased.
Vegetative growth of P. vulgaris was lowered at greater concentrations of PME. A high
EC indicates greater salt content in the greater PME concentrations, which decreased the
shoot length, root length, dry weight, chlorophyll content, and LAI plant−1of P. vulgaris.
Vegetative growth is associated with development of new shoots, twigs, leaves, and leaf
area. Shoot length, root length, dry weight, and LAI plant−1of P. vulgaris were greater
at 25% of PME. This may be due to maximum uptake of N, P, and K by plants. The
improvement of vegetative growth may be attributed to the role of K in nutrient and sugar
translocation in plants and turgor pressure in plant cells. It is also involved in cell enlarge-
ment and in triggering young tissue or mersitematic growth (Sundri and Kanakarani 2001).
Chlorophyll content was greater due to use of 25% PME in both seasons and is likely due
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Ferti-irrigation Effect of Paper Mill Effluent 2163
Table 9
Coefficient of correlation (r) between PME and P. vulgaris in both seasons
PME P. vulgaris Season r value
PME versus shoot length Rainy +0.66
Summer +0.65
PME versus root length Rainy +0.13
Summer −0.14
PME versus dry weight Rainy +0.23
Summer +0.38
PME versus chlorophyll content Rainy +0.27
Summer +0.27
PME versus LAI Rainy +0.47
Summer +0.50
PME versus no. of flowers plant−1Rainy +0.59
Summer +0.27
PME versus soil no. of pods (I-H) Rainy +0.58
Summer +0.41
PME versus no. of pods (II-H) Rainy +0.48
Summer +0.32
PME versus no. of pods (III-H) Rainy +0.27
Summer +0.27
PME versus CY plant−1(fresh) Rainy +0.10
Summer −0.06
PME versus CY plant−1(dry) Rainy +0.39
Summer +0.31
PME versus HI Rainy +0.47
Summer +0.01
PME versus Cd Rainy +0.99
Summer +0.99
PME versus Cr Rainy +0.95
Summer +0.96
PME versus Cu Rainy +0.99
Summer +0.99
PME versus Mn Rainy +0.98
Summer +0.98
PME versus Zn Rainy +0.99
Summer +0.99
to Fe, Mg, and Mn contents in the PME, which are associated with chlorophyll synthe-
sis (Porra 2002). The 25% PME concentration contains optimum contents of nutrients
required for maximum vegetative growth of P. vulgaris.
Effect on Flowering
Season, PME concentration, and interaction of season and PME concentration had no effect
on number of flowers and number of pods/plant (Table 10). At flowering stage (60 days
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2164 V. Kumar and A. K. Chopra
Table 10
ANOVA for effect of PME on flowering and maturity stage of P. vulgaris
Source
No. of
flowers
plant−1
No. of
pods
(I-H)
No. of
pods
(II-H)
No. of
pods
(III-H)
CY
plant−1
(fresh)
CY
plant−1
(dry) HI
Season (S) ns ns ns ns ns ns ns
PME concentration (C) ns ns ns ns ns ns ns
Interaction S ×C nsnsnsnsnsnsns
Notes. ns, nonsignificant; I-H, II-H, and III-H denote harvest I, harvest II, and harvest III,
respectively.
after sowing) the most flowers (40.00 and 38.00) were with 25% PME in both seasons.
Numbers of flowers decreased as PME concentration decreased (Table 9). Numbers of
flowers plant−1of 30.00 and 32.00 were found in the control and 35.00 to 37.00 with
100% PME in both seasons. Again, for all the parameters the lower value is for the rainy
season and the higher value is for the summer season.
Nitrogen and P are essential for flowering. Too much N can delay or prevent flower-
ing whereas P deficiency is sometimes associated with poor flower production, or flower
abortion. Maximum flowering was with the 25% PME; it might indicate that this concen-
tration contains sufficient N and P. Furthermore, P and K prevent flower abortion so pod
formation occurs (El-Naggar 2005). Flowering of P. vulgaris was lower at greater concen-
trations of PME. This is likely due to increased contents of metals in the soil, which inhibit
uptake of P and K by plants at greater PME concentrations (Pandey, Nautiyal, and Sharma
2008).
Effect on Maturity
Numbers of pods/plant, crop yield/plant, and harvest index (HI) were not affected by
season, PME concentration, and their interaction (Table 10). Pods/plants at harvests I
(10.00 and 11.50), II (12.00 and 12.70), and III (8.00 and 8.90), fresh yield plant−1
(215.60 and 220.40 g), dry yield plant−1(40.30 and 43.50 g), and HI (532.40 and 540.60%)
were with the control while with 100% PME the pods/plants at harvests I (14.00 and
15.40), II (15.0 and 15.60), and III (10.30 and 11.40), fresh yield plant−1(228.00 and
232.00 g), dry yield plant−1(52.00 and 58.00 g), and HI (575.00 and 580.00%) were in
both seasons.
The most pods/plants at harvests I (17.00 and 18.00), II (19.00 and 20.60), and III
(15.00 and 15.90), fresh yield/plant (280.70 and 396.60g), dry yield/plant (75.60 and
85.80 g), and HI (612.70 and 625.00%) were with the 25% PME in both seasons (Table 6).
Numbers of pods/plant, crop yield plant−1, and harvest index (HI) of P. vulgaris were
positively correlated with PME concentrations in both seasons (Table 9). The roles of K,
Fe, Mg, and Mn at maturity are important, associated with synthesis of chlorophyll, and
enhance formation of pods at harvest (El-Naggar 2005; Naeem, Iqbal, and Bakhsh 2006).
The K, Fe, Mg, and Mn contents could benefit pod formation and yield as it does for
soybean (Glycine max L.), as reported by Hati et al. (2007). The 25% PME favored pod
formation and crop yield of P. vulgaris. This is likely due to presence of K, Fe, Mg, and
Mn contents in 25% PME; greater PME concentrations lowered pod formation and crop
yield of P. vulgaris.
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Ferti-irrigation Effect of Paper Mill Effluent 2165
Effect on Biochemical Constituents and Micronutrients in P. vulgaris
Season, PME concentration, and the interaction of season and PME concentration affected
all the biochemical constituents such as crude fiber and crude carbohydrates and metals
such as Cd, Cr, Cu, Mn, and Zn in P. vulgaris (Table 11). Maximum crude proteins, crude
fiber, and crude carbohydrates were recorded with 25% PME concentrations in both sea-
sons (Figures 4–6). Content of crude proteins (r =+08 and r =+0.14), crude fiber (r =
+0.14 and r =+0.08), and crude carbohydrates (r =+0.13 and r =+0.08) were noted
positively correlated with PME concentration in both seasons. The 25, 50, 75, and 100%
PME concentrations affected Cd, Cr, Cu, Mn, and Zn contents in P. vulgaris. Increased
irrigation frequency could lead to increases of metals in tissues. The Cd, Cr, Cu, Mn, and
Zn contents in P. vulgaris were greatest with 100% PME (Figures 7 and 8). They were
correlated with concentrations of Cd, Cr, Cu, Mn, and Zn in P. vulgaris and positively
correlated with PME concentrations in both seasons (Table 9).
The contamination factor (Cf) was affected in both seasons (Figure 9). The Cf of
various metals was in order of Mn >Zn >Cu >Cd >Cr in P. vulgaris after irrigation
with PME (Figure 9). The greatest contamination factor was for Mn; the least was for Cr
in P. vulgaris with 100% PME in both seasons. The micronutrient contents were greater
at greater PME concentration and likely inhibited growth of P. vulgaris. The 25% PME
favored vegetative growth, flowering, and maturity of P. vulgaris. This is likely due to
Table 11
ANOVA for effect of PME on concentrations of metals in P. vulgaris
Source Cd Cr Cu Mn Zn
Crude
proteins
Crude
fiber
Crude
carbohydrates
Season (S) ∗∗∗
ns ∗∗ ∗ ∗
PME concentration (C) ∗∗ ∗∗ ∗∗ ∗∗ ∗∗ ∗∗ ∗∗ ∗∗
Interaction S ×C∗∗ ∗∗ ∗∗ ∗∗ ∗∗ ∗∗ ∗∗ ∗∗
Notes. ns, ∗,and∗∗ denote nonsignificant or significant at P≤0.05 or P≤0.01 by ANOVA,
respectively.
0
5
10
15
20
25
0 5 10 25 50 75 100
PME concentration (%)
Crude proteins (%)
Crude proteins in rainy season
Crude proteins in summer season
Figure 4. Crude proteins in P. vulgaris after irrigation with PME. Error bars are standard error of
the mean.
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2166 V. Kumar and A. K. Chopra
0
5
10
15
20
25
30
35
40
45
50
1234567
PME concentration (%)
Crude fiber (%)
Crude fiber in rainy season
Crude fiber in summer season
Figure 5. Crude fiber in P. vulgaris after irrigation with PME. Error bars are standard error of the
mean.
0
2
4
6
8
10
12
14
16
18
20
0 5 10 25 50 75 100
PME concentration
(
%
)
Crude carbohydrates (%)
Crude carbohydrates in rainy season
Crude carbohydrates in summer season
Figure 6. Crude carbohydrates in P. vulgaris after irrigation with PME. Error bars are standard error
of the mean.
0
1
2
3
4
5
6
7
0 5 10 25 50 75 100
PME concentration (%)
Metal content (mg kg–1)
Cd in rainy season Cd in summer season
Cr in rainy season Cr in summer season
Cu in rainy season Cu in summer season
Figure 7. Contents of Cd, Cr and Cu in P. vulgaris after irrigation with PME. Error bars are standard
error of the mean.
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Ferti-irrigation Effect of Paper Mill Effluent 2167
0
0.5
1
1.5
2
2.5
3
3.5
4
4.5
5
0 5 10 25 50 75 100
PME concentration (%)
Metal content (mg kg–1)
Mn in rainy season Mn in summer season
Zn in rainy season Zn in summer season
Figure 8. Contents of Mn and Zn in P. vulgaris after irrigation with PME. Error bars are standard
error of the mean.
0
1
2
3
4
5
6
7
8
Zn Mn Cd Cu Cr
Metals
Contamination factor (Cf)
Cf in rainy season
Cf in summer season
Figure 9. Contamination factors of various metals in P. vulgaris after irrigation with PME. Error
bars are standard error of the mean.
optimal uptake of these micronutrients by crop plants, which supports various biochemical
and physiological processes.
Conclusion
In conclusions, paper mill effluent increased nutrients in the soil and affected the growth
of P. vulgaris in both seasons. The effluent has potential for use as biofertigant in the
form of plant nutrients needed by P. vulgaris crop plant. Therefore, it can be used as agro-
based biofertigant after its appropriate dilution for irrigation purposes for the maximum
yield of this crop. Further studies on the agronomic growth and changes in biochemical
composition of P. vulgaris after PME irrigation are required.
Funding
The University Grants Commission, New Delhi, India, is acknowledged for providing the
financial support in the form of a research fellowship (F.7-70/2007-2009 BSR) to the
corresponding author.
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2168 V. Kumar and A. K. Chopra
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