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ABSTRACT In recent years integrated system of composting, with bioinoculants and subsequent vermicomposting, to overcome the problem of lignocellulosic waste degradation of different crop residues and waste industrial
by-products is receiving worldwide attention of scientists. However, there appears no report available on the technical viability of the approach on waste
by-products of sugar-cane industry. Therefore, in the present study, the waste
by-products of sugar-cane industry viz. bagasse (b), press-mud (p), and trash
(t) have been subjected to bioinoculation followed by vermicomposting with
a view to shorten stabilization time, and improve the product quality. The
substrate (press-mud alone and in combination of other by-products of sugarprocessing industries) was predecomposed for 30 days by inoculating it with
Pleurotus sajorcaju, Trichoderma viridae, Aspergillus niger, and Pseudomonas striatum
in different combinations. This was followed by vermicomposting for 40 days
using Drawida willsi species of the native earthworms. Results indicated that
the combinations of both the systems reduced the overall time required for
composting to 20 days and accelerated the degradation process of waste byproducts of sugar-processing industry, thereby producing a nutrient-enriched
compost product.
KEYWORDS bioinoculants, Drawida willsi, integrated system of composting, microbial
activity, sugar-cane industries, vermicompost
INTRODUCTION
Address correspondence to Shweta,
Vermiculture Research Station,
Department of Zoology, Dharam
Samaj College (Dr. B. R. A. University),
Aligarh, 202001, India. E-mail:
kmshweta3@yahoo.com
of Economics and Statistics, India produces on an average 270 million tons of sugar cane per year. During
the production process considerable amounts of byproducts such as press-mud, bagasse, and sugar-cane
trash are produced. Part of these by-products can be
utilized for the production of molasses and alcohol;
however, there still remains a considerable amount of
waste products to be disposed. Therefore, there is considerable economic interest in the technology and development processes for effective utilization of these
wastes (Zhang, 2000). As a result, emphasis is now on
aerobic composting, which converts wastes into organic
manure rich in plant nutrients and humus (Singh and
Sharma, 2002). Even though recycling organic wastes
has been known since biblical times, yet there are many
aspects that needs to be improved. One of these aspects
includes reduction in overall time required for composting. Various studies have shown that by-products
of sugar-cane processing are ideal substrates for breeding of earthworms (Pramanik et al., 2007; Manna et al.,
2003) and gives a product rich in chelating and phytohormonal elements that has a high content of microbial agents and stabilized humic substances (Atiyeh
et al., 2001). Furthermore, combining vermicomposting with composting also accelerated the composting
process, thus reducing the time required for composting (Frederickson et al., 1997; Nedgwa and Thompson,
2001). But because some epigeic earthworm species
require predecomposed waste to be vermicomposted,
it would be desirable to decrease the predecomposition time period of the waste initially with certain efficient microflora. Lignin is the most recalcitrant material present in by-products of sugar industries and
decomposes only at the later stage of decomposition
(Manna et al., 2003). The rate of decomposition could
be enhanced by treating the wastes initially with certain efficient microflora (Singh and Sharma, 2002).
Pleurotus sajorcaju, Trichoderma viridae, and Aspergillus
niger are known for degrading hemicelluloses and cellulose, respectively. These fungi, which produce cellulase enzyme, were used as effective inoculants for
composting (Banitez et al., 2000). However, the inoculation of phosphate-solublizing bacteria Pseudomonas
striata may help to solubilize phosphorus and to increase its availability to plants (Eilan, 2001). Therefore,
these microflora could be used as inoculants during
predecomposition of the wastes, which may reduce the
time of composting. The present work is based on this
rationale.
159
METHODS
Microbial Source
The fungus strains P. sajorcaju, A. niger, and T. viridae were procured from IMTECH, Chandigarh, and
the bacterial strain P. striatum was obtained from Indian Agricultural Research Institute, New Delhi. The
fungal cultures were maintained by subculturing them
on potato dextrose agar whereas bacteria was subcultured on Pikovaskaya medium.
Experimental Setup
Press-mud ( p) alone and in combination with
bagasse (b) and sugarcane trash (t) were used as substrate for the predecomposition studies. Finely chopped
substrate was pasteurized by dipping overnight in 0.1%
formalin. Predecomposition of the sterilized industrial
wastes of sugar mills was done in pits (1 m 1 m
1 m). The experiments were conducted with 60 kg of
substrate during April to June 2009. Pure cultures of
P. sajorcaju, T. viridae, and A. niger (500 g mycelium
per ton substrate) and P. striatum (50 ml/kg substrate
having 106 cells per ml) were inoculated in different
combinations:
P. sajorcaju (P)
P. sajorcaju + T. viridae (PT)
P. sajorcaju + T. viridae + A. niger (PTA)
P. sajorcaju + T. viridae + A. niger + P. striata (PTAS)
For mesophilic aerobic digestion, turning was done
manually every 4 days and the temperature was not
allowed to exceed 26 C. The substrate with different
treatments was predecomposed, in triplicates, for 30
days and then subjected to vermicomposting for 40
days.
Composting and Vermicomposting of Sugar-Cane By-products
TABLE 1 Chemical Analysis of Sugar-Cane Waste By-products (Press-Mud Alone and in Combination)
Substrate
Parameter
Total organic carbon (TOC)
Total Kjeldahl Nitrogen (TKN)
Total Phosphorus (TP)
Total Potassium (TK)
Cellulose
Hemicellulose
Lignin
Pressmud (p)
Press-mud +
sugarcane trash (ps)
Press-mud +
bagasse (pb)
Press-mud + sugarcane
trash + bagasse (psb)
55.06 0.10
0.85 0.19
0.53 0.04
1.72 0.01
62.69 0.01
23.07 0.02
10.07 0.02
51.21 0.20
0.80 0.10
0.58 0.04
1.28 0.01
68.72 0.01
24.07 0.01
22.37 0.02
56.05 0.20
0.82 0.20
0.53 0.02
1.60 0.01
65.73 0.02
25.07 0.01
16.70 0.02
56.07 0.01
0.81 0.01
0.55 0.02
1.62 0.01
68.69 0.02
26.71 0.02
18.37 0.01
Note. All values are given in percentage. All values are mean of three replicates.
Vermiculturing
Statistical Analysis
For subsequent vermicomposting of the predecomposed waste, the earthworms Drawida willsi Michaelsen
were cultured in cow dung employing the windrows
method.
All the results reported are the means of three replicates. One-way analysis of variance (ANOVA) was done
using the INDOSTAT program. The objective of statistical analysis was to determine any significant differences among the parameters analyzed in different
treatments during the composting process.
Vermicomposting
The predecomposed substrates were vermicomposted, in the same pits as used for predecomposition,
for a period of 40 days. Moisture was maintained to
about 60% of the water holding capacity, 50 earthworms (Drawida willsi) were introduced, in each pit,
in the predecomposed sugar-processing industries byproducts, viz. press-mud ( p), press-mud + sugarcane
trash 1:1 (ps), press-mud + bagasse 1:1 (pb), press-mud
+ sugarcane trash + bagasse 1:1:1 (psb), and treated with
different combinations of bioinoculants. Sampling was
done at an interval of 10 days. Composite samples
(about 100 g) were collected from three sites in each
pit. The earthworms and cocoons were removed manually and vermicompost was chemically analyzed.
Compost Analysis
Total Kjeldahl nitrogen (TKN) and total organic
carbon (TOC) of the predecomposed bioinoculated
residue and the vermicompost were estimated by using
micro-Kjeldahl method (Singh and Pradhan, 1981) and
Walkey and Blacks rapid titration method (1934), respectively. Total phosphorus (TP) was determined spectrophotometrically whereas total potassium (TK) was
detected by the flame emission technique. Cellulose,
hemicellulose, and lignin were fractionated sequentially
by Duttas method (1981) (Table 1).
Shweta et al.
161
20 days
22.86 0.19*
26.36 0.02*
25.12 0.10*
27.14 0.01*
24.00 0.10*
21.36 0.19*
26.20 0.01*
27.18 0.19*
27.79 0.20*
27.00 0.17*
20.76 0.17*
26.12 0.16*
27.02 0.11*
28.00 0.12*
28.02 0.11*
24.86 0.12*
25.20 0.20*
26.78 0.04*
29.28 0.01*
29.26 0.01
10 days
14.86 0.12
17.86 0.52*
18.96 0.12*
18.20 0.10*
18.01 0.11*
12.36 0.10
14.86 0.10*
17.96 0.07*
18.20 0.20
18.20 0.24*
14.20 0.20
16.86 0.21*
17.86 0.10
17.90 0.14*
17.80 0.04
15.98 0.02
17.90 0.06*
18.30 0.12*
19.28 0.02*
19.20 0.01*
14.30 0.18
18.30 0.05*
18.32 0.16*
18.76 0.05*
18.76 0.05*
15.30 0.16
18.20 0.15*
19.00 0.10*
19.89 0.10*
19.90 0.11*
15.30 0.17
17.30 .01*
17.86 0.31*
18.01 0.20*
18.98 0.01*
14.60 0.01
18.78 0.01
19.12 0.07*
20.01 0.02*
20.01 0.02*
30 days
Significant
Note. All values are mean and standard deviation of three replicates
( p < .01).
p = press-mud; s =sugar-cane trash; b = bagasse; P = Pleurotus sajorcaju; T = Trichoderma viridae; A = Aspergillus niger; S = Pseudomonas striatum.
p (control)
p+ P
p+ PT
p+ PTA
p+ PTAS
ps (control)
ps + P
ps + PT
ps + PTA
ps + PTAS
pb (control)
pb + P
pb + PT
pb + PTA
pb + PTAS
psb (control)
psb + P
psb + PT
psb + PTA
psb + PTAS
Treatment
15.30 0.01
15.30 0.18
14.39 0.05
13.20 0.05
13.68 0.05
14.30 0.01
16.60 0.18*
17.40 0.05*
16.42 0.06*
16.88 0.05*
15.60 0.01
14.39 0.05
14.00 0.06
13.02 0.15
13.62 0.02
12.65 0.15
13.60 0.02
13.24 0.12
12.62 0.03
12.00 0.04
70 days
TABLE 2 Percentage of Decrease in Total Organic Carbon (TOC) in Sugar-Cane Waste By-products (Press-Mud Alone and in Combination) during Microbial Predecomposition
(030 Days) and Subsequent Vermicomposting (70 Days)
162
20 days
0.95 0.20
1.45 0.20*
1.48 0.25*
1.58 0.10*
1.58 0.15*
0.98 0.10
1.45 0.15
1.45 0.01
1.59 0.10*
1.56 0.15*
0.95 0.20
1.45 0.15*
1.44 0.10*
1.59 0.20*
1.52 0.25*
0.98 0.10
1.52 0.02*
1.56 0.02*
1.61 0.01*
1.60 0.01*
10 days
0.80 0.01
0.84 0.05*
0.83 0.10*
0.84 0.20*
0.84 0.10*
0.80 0.20
0.81 0.10
0.80 0.18
0.84 0.20*
0.84 0.28*
0.81 0.20
0.81 0.01
0.80 0.03
0.84 0.05*
0.83 0.01
0.81 0.01
0.82 0.20
0.83 0.20*
0.84 0.05*
0.84 0.50*
Significant
Note. All values are mean and standard deviation of three replicates.
( p < .01).
p (control)
p+ P
p+ PT
p+ PTA
p+ PTAS
ps (control)
ps + P
ps + PT
ps + PTA
ps + PTAS
pb (control)
pb + P
pb + PT
pb + PTA
pb + PTAS
psb (control)
psb + P
psb + PT
psb + PTA
psb + PTAS
Treatment
0.95 0.20
1.45 0.30*
1.48 0.01*
1.50 0.18*
1.52 0.18*
0.96 0.02
1.45 0.01*
1.45 0.02*
1.42 0.01
1.56 0.02*
0.95 0.01
1.45 0.01*
1.44 0.20*
1.51 0.18*
1.50 0.15*
0.98 0.01
1.56 0.10*
1.57 0.20*
1.62 0.25*
1.60 0.20*
30 days
0.90 0.20
1.42 0.18*
1.42 0.15*
1.40 0.10*
1.40 0.18*
0.92 0.05
1.40 0.05*
1.40 0.10*
1.38 0.10
1.38 0.20
0.93 0.20
1.40 0.02*
1.38 0.02*
1.36 0.01*
1.38 0.01*
0.96 0.02
1.40 0.01
1.42 0.01*
1.40 0.00
1.40 0.02
70 days
TABLE 3 Percentage of Total Kjeldahl Nitrogen (TKN) in Sugar-Cane Waste By-products (Press-Mud Alone and in Combination) during Microbial Predecomposition (030 Days)
and Subsequent Vermicomposting (70 Days)
163
TK
1.68 0.20*
1.70 0.20*
1.74 0.15*
1.74 0.10*
1.74 0.10*
1.68 0.20*
1.70 0.21*
1.70 0.20*
1.74 0.20*
1.76 0.15*
1.65 0.10*
1.78 0.10*
1.78 0.15*
1.78 0.10*
1.81 0.10*
1.68 0.10*
1.72 0.10*
1.74 0.15*
1.74 0.10*
1.80 0.10*
TP
0.53 0.01
0.53 0.02*
0.52 0.01*
0.50 0.02*
0.52 0.01*
0.58 0.01*
0.56 0.02*
0.58 0.15*
0.56 0.10*
0.56 0.01*
0.58 0.02
0.56 0.02*
0.54 0.01*
0.54 0.02*
0.55 0.01*
0.58 0.01
0.59 0.01
0.58 0.02*
0.58 0.01*
0.59 0.01*
Significant
TP
TK
1.80 0.10
1.93 0.10*
1.92 0.01*
1.91 0.02
1.90 0.02
1.80 0.10
1.93 0.15*
1.92 0.10*
1.91 0.10*
1.90 0.12*
1.80 0.15
1.92 0.10
1.91 0.15*
1.93 0.10*
1.93 0.10*
1.74 0.12
1.93 0.10*
1.93 0.10*
1.92 0.15*
1.91 0.10
20 days
0.56 0.10*
0.70 0.15*
0.70 0.10*
0.74 0.15*
0.98 0.10*
0.60 0.15*
0.78 0.10*
0.80 0.12*
0.81 0.12*
0.98 0.10
0.60 0.10*
0.70 0.15
0.78 0.10*
0.78 0.10*
0.98 0.15*
0.62 0.15
0.74 0.10*
0.80 0.10*
0.88 0.10*
0.98 0.15*
Note. All values are mean and standard deviation of three replicates.
( p < .01).
p (control)
p+ P
p+ PT
p+ PTA
p+ PTAS
ps (control)
ps + P
ps + PT
ps + PTA
ps + PTAS
pb (control)
pb + P
pb + PT
pb + PTA
pb + PTAS
psb (control)
psb + P
psb + PT
psb + PTA
psb + PTAS
Treatment
10 days
0.60 0.01
0.74 0.01*
0.72 0.02*
0.76 0.02*
0.98 0.02*
0.68 0.01*
0.80 0.02*
0.82 0.15*
0.84 0.10*
0.98 0.01
0.62 0.02
0.78 0.02*
0.82 0.01*
0.82 0.01*
0.98 0.02*
0.63 0.01*
0.84 0.01*
0.92 0.02*
0.92 0.01*
0.99 0.01*
TP
TK
1.88 0.20
1.96 0.20
1.96 0.15
1.98 0.15*
1.96 0.20
1.82 0.25
1.96 0.10
1.96 0.25
1.92 0.25
1.96 0.20*
1.82 0.25
1.96 0.20*
1.98 0.15*
1.98 0.10*
1.97 0.10*
1.81 0.10
1.92 0.15*
1.93 0.20
1.94 0.21
1.96 0.20
30 days
0.70 0.20
1.20 0.20*
1.20 0.15*
1.28 0.10*
1.78 0.15*
0.84 0.10*
1.28 0.20
1.32 0.20*
1.32 0.15*
1.88 0.10*
0.80 0.15
1.20 0.10
1.20 0.15
1.34 0.10*
1.98 0.15*
0.88 0.10
1.24 0.15
1.25 0.20
1.24 0.25
1.88 0.20*
TP
70 days
1.62 0.20
1.80 0.20*
1.88 0.18*
1.84 0.10*
1.84 0.15*
1.60 0.10
1.80 0.15*
1.80 0.15*
1.78 0.20*
1.81 0.20*
1.62 0.15
1.80 0.10*
1.80 0.15*
1.81 0.15*
1.82 0.15
0.60 0.10
1.82 0.10*
1.80 0.10*
1.78 0.15
1.78 0.10
TK
TABLE 4 Percentage of Total Phosphorus (TP) and Total Potassium (TK) in Sugar-Cane Waste By-products (Press-Mud Alone and in Combination) during Microbial Predecompo-
164
70 days
42.63 0.49
30.46 0.10*
24.12 0.18*
20.18 0.25*
22.32 0.20*
44.12 0.41*
28.16 0.41*
23.39 0.20
18.73 0.20*
17.64 0.25*
40.26 0.25*
22.62 0.20*
20.62 0.40*
16.18 0.48*
17.60 0.20
48.64 0.48*
24.67 0.40
20.60 0.40*
16.73 0.45*
16.20 0.40*
30 days
60.69 1.17
45.63 1.10*
41.12 1.00 *
40.02 1.17*
40.02 1.09*
68.72 1.17
56.72 1.02*
44.60 1.02*
41.12 0.41*
41.02 0.40*
65.73 1.02*
57.34 1.17*
45.70 1.10*
43.22 1.02*
42.12 1.12*
68.69 1.10
54.18 1.12*
45.70 1.02*
38.22 1.12*
38.30 1.02*
Significant
Note. All values are mean and standard deviation of three replicates.
( p < .01).
p (control)
p+ P
p+ PT
p+ PTA
p+ PTAS
ps (control)
ps + P
ps + PT
ps + PTA
ps + PTAS
pb (control)
pb + P
pb + PT
pb + PTA
pb + PTAS
psb (control)
psb + P
psb + PT
psb + PTA
psb + PTAS
Treatment
Cellulose (%)
33.07 0.32
22.35 0.18*
21.60 0.20*
18.62 0.25*
20.62 0.20*
35.67 0.41
20.35 0.41*
20.60 0.40*
17.60 0.45*
17.22 0.40*
34.62 0.40
22.32 0.45*
21.65 0.40*
21.22 0.45*
21.22 0.40*
38.67 0.40*
24.25 0.45*
21.20 0.20*
20.13 0.20*
20.10 0.25*
30 days
24.56 0.32
19.15 0.28*
18.97 0.49*
15.54 0.58*
15.50 0.50
28.60 0.58*
18.16 0.48*
17.27 0.40
15.25 0.48*
15.10 0.40*
25.32 0.07
18.16 0.41
16.24 0.40*
14.37 0.41*
14.30 0.48*
29.61 0.49
20.26 0.48*
18.28 0.49*
16.24 0.40*
16.20 0.40
70 days
Hemicellulose (%)
14.16 0.38
11.56 0.40*
10.91 0.45*
10.53 0.48*
10.50 0.45
15.36 0.40
12.16 0.42*
10.91 0.40*
10.51 0.48*
10.00 0.40*
14.12 0.19
11.62 0.15*
10.60 0.15*
10.58 0.20*
10.32 0.28*
15.16 0.38*
13.12 0.49*
12.10 0.40*
11.18 0.40*
10.01 0.48
30 days
Lignin (%)
70 days
8.76 0.20
5.13 0.25*
3.60 0.38*
3.20 0.40*
3.00 0.45
9.26 0.48
5.42 0.58*
3.67 0.49*
3.18 0.45*
3.20 0.48
8.80 0.50*
5.18 0.58*
3.52 0.20*
3.23 0.20
2.81 0.25
9.26 0.29*
5.10 1.15*
3.52 0.25*
3.42 0.25
3.20 0.28
TABLE 5 Percentage of Cellulose, Hemicellulose, and Lignin Contents in Predecomposed and Subsequent Vermicomposted Waste of Sugar-Cane Waste By-products
significantly both during predecomposition and subsequent vermicomposting, with maximum degradation
with all four bioinoculants (PTAP) (Table 5). Similar results were also reported by Singh and Sharma
(2002) who reported rapid decomposition of wheat
straw with a mixture of celluloytic fungi, Pleurotus
sajorcaju, Trichoderma reesei, and Aspergillus niger, along
with the nitrogen-fixing bacteria Azatobacter chroococcum. Along with this, the simultaneous activity of microflora present in the gut of earthworms and in the
waste substrate might have intensified cellulolysis and
lignolysis as suggested by Loquiet et al. (1984). The
structure of lignin changes, probably due to microbial
oxidation and demethylation. The microbial cleavage
of the aromatic rings of lignin leads to new polysaccharide and humus in the organic matter (Beyer et al.,
2005).
A noticeable increase in the number of earthworms
as well as the cocoons was observed during vermicomposting (Table 6). This increase in the growth of
earthworms with phosphorus-solubilizing bacteria Pseudomonas striatum suggests the dual role of bacteria, i.e.,
in having been utilized as food material and enriching the substrate with phosphorus through phosphorus
TABLE 6 Changes in Growth and Development of Earthworms
(Drawida willsi) during Vermicomposting of Sugar-Cane Waste
By-products (Press-Mud Alone and in Combination)
Treatment
p (control)
p+ P
p+ PT
p+ PTA
p+ PTAS
ps (control)
ps + P
ps + PT
ps + PTA
ps + PTAS
pb (control)
pb + P
pb + PT
pb + PTA
pb + PTAS
psb (control)
psb + P
psb + PT
psb + PTA
psb + PTAS
Total earthworm
Cocoon
89.0 2.00
118 4.00*
121 4.00*
128 8.00*
128 8.85*
92.6 2.16
119 4.16*
124 4.80*
128 6.56*
130 7.56*
87.6 2.12
118 4.81*
121 6.80*
128 6.00*
128 4.02*
93.2 4.06
128 4.58*
128 3.01*
131 3.80*
133 3.01*
20 2.01
35 3.58*
39 3.58*
42 3.62*
47 2.55
22 4.88
35 2.00*
38 3.00*
41 4.05*
42 4.05*
20 6.05
38 4.08*
39 3.05*
42 4.05*
42 2.05*
24 3.08
39 4.80*
40 5.08*
44 4.08*
44 3.80*
Note. All values are mean and standard deviation of three replicates.
( p < .01).
Significant
CONCLUSION
The chemical analyses of the compost produced by
predecomposing of press-mud with other by-products
of sugar-processing industries, with efficient microbes
followed by vermicomposting point towards the feasibility of integrated system of vermicomposting process
tested in the present study. From the lignin degradation
point of view, the results suggest that this integrated system would be best for lignocellulosic waste treatment of
sugar caneprocessing industry. Reduction in the predecomposition time that is from 40 to 20 days would
enable us to potentially convert these sugar-cane byproduct wastes into value added products in a short
time.
ACKNOWLEDGMENT
The authors are thankful to the Department of
Biotechnology, Ministry of Science and Technology,
Government of India, New Delhi, for financial assistance.
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