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Vermicomposting of de-oiled cake of Brassica juncea by Eisenia foetida.

Rapeseed-mustard is an important oil seed crop and grown on 34.19 million hectares area with a production of63.09 million tons in the world during 2012-13. In India it covered 19.29% acreage area and 11.127% of production in this period (http:/ /www.drmr.res.in/about_rm.html, retrieved on 12/ 1/2015). The oil seeds contain Glucosinolates. When oil seeds are processed to remove oil these glucosinolates converted into thiocyanates, nitriles, isothiocyanates, epithionitriles etc. by the action of myrosinases. These products are toxic to mammalian species because they interfere with uptake of iodine causing goiter. Its de-oiled cake is commonly used as cattle feed but it is not safe due to above facts. Harmful effects of glucosinolates are reported by many workers (Duncan, 1991; Barrett et al, 1997; Papas et al, 1979; Fenwick and Cruits, 1980). Therefore we tried a different approach for management of this waste through vermicomposting. Earthworm's gizzard has numerous of enzymes having capacity to rupture a number of complex molecules.

Vermicomposting is a proven technology to convert a variety of plant wastes into a best fertilizer which supports plant growth (Benitez et al., 1999; Atiyeh et al., 2002). Eisenia foetida species of earthworm is widely accepted for vermicomposting of agricultural waste.

MATERIALS AND METHODS

In the present experiment we used de-oiled cake of Mustard (Brassica juncea), earthworms species Eisenia foetida and buffalo dung. Mustard cake was obtained from Kukarvada, Dist: Mehsana and other materials were procured locally from sadra and Randheja village of District Gandhinagar, Gujarat. Before addition to vermicompost pits the cake was chopped into small pieces (about 1 inches long). For vermicomposting pits from bricks were prepared on above ground of our campus. These pits were 45cm long, 45cm wide and 21cm high.

Treatments

Above materials were mixed in different combinations to obtain four treatments. Control comprises Buffalo dung and Eisenia foetida only; Treatment T1 comprises Buffalo dung and Brassica juncea cake without earthworms whereas treatment T2 contains earthworms alongwith Buffalo dung and Brassica juncea cake. Last treatment T3 contains Brassica juncea cake and Eisenia foetida alone without dung. To minimize the experimental error all the above treatments were replicated thrice.

Filling of pits

A total of six kilogram of material was filled in each pit. In pits of control and T3 treatments 6kg buffalo dung and Brassica juncea cake were filled alone whereas in T1 and T2 the material was filled in layers. In layered treatments a layer of 3Kg Buffalo dung was kept at bottom of pit followed by one layer of 1Kg mustard cake which was covered by 2Kg of buffalo dung. Fermenting material was kept moist and turned after every seven days. Earthworms in respective pits were added after second turning in the amount of 18g except T1. To prevent moisture loss all pits were covered with gunny bags. The material was analysed at 30, 60 and 90 days of filling to find out the changes in various chemical properties. At maturity (90 days) the Earthworms were removed from pits and samples were analyzed for various parameters along with yield.

Analysis of vermicomposting material

The samples at different time intervals were analysed for physico-chemical parameters including pH and electrical conductivity (Richard's, 1945), total organic carbon (Walkely and Black, 1965), nutrients like phosphorus, potassium and sodium (Jackson, 1967), and for cellulose, hemicellulose and lignin content (Thimmaiah, 1999). FTIR (Fourier-Transform Infrared Spectroscopy) analysis was also performed at Center of Excellence, Vapi, Gujarat.

Statistical analysis

Completely Randomized Block Design was followed and significance level was find out to check the significance in obtained data at 5% level following procedure described in Gomez and Gomez, (1984).

RESULTS AND DISCUSSION

Yield of vermicompost

During vermicomposting earthworm ingest the substrate, digest it, absorbs nutrients and discard undigested part as castings. If the substrate is easy to digest, contains simple forms, then more nutrients will be absorbed and eventually less part will be discarded with lower yield of product. In present experiment two substrates were used viz. buffalo dung and mustard cake. Compared to mustard cake buffalo dung is easy to degrade because it is a partially degraded product of buffalo so it contains simple forms of compounds which can be digested easily by earthworm's gut flora. Due to this fact we would obtain lesser amount of product in this case. Our results are in tune with this fact and lowest yield is recorded in control treatment containing buffalo dung and earthworms (Table 1). As a portion of buffalo dung is replaced by mustard cake ([T.sub.1] and [T.sub.2]) yield of vermicompost increased significantly and highest yield was obtained with mustard cake alone ([T.sub.3]) which differs significantly from rest of the treatments (Table 1). Weight loss of vermicomposting material at final stage was also reported by Twana and Fauziah, 2012; Senthilkumar and Kavimani, 2012.

Chemical properties

Data presented in Table 2 and Table 3 reveals that pH, electrical conductivity, total organic carbon, available P, K and Na contents varies with treatments and except organic carbon, contents of all other parameters increased at maturity than that of initial. Further data show that at various stages of analysis the effect of treatments was significant and at some stages it was non-significant. At maturity of vermicompost the effect of treatments was found significant for pH, available phosphorus and sodium. Intestinal calcium secretion and excretion of ammonia by earthworms raise the pH of vermicompost and make it alkaline (Ismail, 1997; Ismail, 2005; Edwards and Bohlen, 1996).

Mineralization of organic substrates like cellulose, hemicellulose, lignin etc. during vermicomposting by the activities of earthworms and other microorganisms of dung releases mineral salts (Gupta and Garg, 2008; Khawairakpam and Bhargava, 2009; Kaviraj and Sharma, 2003) which results into increased values of electrical conductivity (Neuhauser et al, 1988). Organic carbon is being utilized by microorganisms as energy source and the respiratory activities of earthworms and microorganisms may results in reduced amount of organic carbon in final product (Curry et al, 1995).

Available phosphorus, potassium and sodium

Microbial mineralization of organic phosphorus and activities of earthworms gut phosphatase resulted in increased amount of available phosphorus in product (Tripathi and Bharadwaj, 2004; Lee, 1992). Acid production is claimed during microbial degradation (Premuzic et al., 1998) which causes solubilization of bound phosphorus, sodium and potassium. Mineralization, transformation and mobilization of nutrients from unavailable forms to plant available forms is being supported by a number of workers (Suthar, 2007; Adi and Noor, 2009; Lazcana et al., 2008; Sen and Chandra, 2009; Ghosh et al., 1999). Solubilization of insoluble potassium due to acid production (Adi and Noor, 2009) and increased amount of potassium in vermicompost due to higher mineralization rate of microbial activity is being well documented (Delgado et al., 1995; Benitez et al., 1999; Kaviraj and Sharma, 2003; Suthar, 2007).

Lignin, hemicellulose, cellulose

Cellulosic material present in substrates are decomposed by microorganisms and earthworms by secreting extracellular enzymes (Sinsabaugh et al., 1992). Digestion of cellulose due to cellulolytic activity in the gut of earthworms and cellulose loss in product is well documented (Lattaud et al., 1997a; Lattaud et al., 1997b; Scheu, 1993; Urba'sek and Pizil, 1991; Vinceslas-Akpa and Loquet, 1997; Zhang et al, 2000; Zhang et al, 1993). Results of present investigation are in line with above workers and showed significant decrease in amount of cellulose and hemicellulose at maturity of vermicompost compared to their initial contents (Table 4).

FTIR (Fourier Transform Infrared Spectrometer)

To prove the degradation of substrate during vermicomposting FTIR data were obtained for a wave number range of 4000-450 [cm.sup.-1]. and presented in Fig 1-7 and details of corresponding groups present is given in Tables 5-8. When Infrared radiation is passed through sample, a portion is absorbed by the sample and remaining is transmitted. This absorption and transmittance spectrum over a wide range of wave number creates a molecular fingerprinting because each molecule gives peak at a specific wave number. Obtained peaks are than compared with spectra library of pure substances or components (Smidt and Schwanninger, 2005). Use of FTIR is very common to describe composting process (Huang et al., 2006, Smidt et al., 2005; Tseng et al., 1996). A close examination of FTIR data obtained for Buffalo dung compared with buffalo dung + Eisenia foetida (T5, Fig.1-2); Brassica juncea compared to Brassica juncea + Eisenia foetida (T-6, Fig. 3-4); Brassica juncea + dung compared with Composted Brassica juncea +dung (T-7 and Fig. 5-6) and Composted Brassica juncea +dung+ Eisenia foetida (T-8, Fig. 7) clearly reveal that they are different in presence of compounds or groups which show that during vermicomposting some groups or compounds are lost and some are newly formed, hence degradation occurs.

CONCLUSION

Results of present investigation clearly direct that de-oiled cake of Brassica juncea can be successfully converted into nutrient rich vermicompost through the activities of Eisenia foetida species of earthworms. In 90 days, biomass, total organic carbon, cellulose, and hemicellulose are reduced by 36.67 to 73.33%, 8.46 to 18.94%, 38.52 to 63.89%, and 11.59 to 45.02%; whereas available phosphorus, potassium and sodium are increased by 64 to 84.85%, 75.44 to 92.00%, and 70.83 to 96.67% respectively. Further the FTIR data confirm the degradation process.

ACKNOWLEDGEMENTS

The experiment was carried out at Department of Biogas Research Center and Department of Microbiology, Gujarat Vidyapith, Sadara, District- Gandhinagar using its laboratory facility hence the help received from the Head of Department is acknowledged.

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Vikram C. Solanki * and Prateek Shilpkar

Biogas Research and Extension Centre and Department of Microbiology, Gujarat Vidyapith, At & Po--Sadara, District--Gandhinagar, Gujarat--382320, India.

(Received: 23 January 2016; accepted: 03 March 2016)

* To whom all correspondence should be addressed. Tel.: +91 9913443451; E-mail: vcsolankicr@gmail.com

Caption: Graph No. : 4.1 FT- IR analysis of Buffalo dung only

Caption: Graph No. : 4.3 FT- IR analysis of Brassica juncas * Eisenia foetida

Caption: Graph No. : 4.4 FT- IR analysis of Brassica juncea only

Caption: Graph No : 4.5 FT- IR analysis of Srassica/uncea ? Buffalo dung

Caption: Graph No : 4.6 FT- IR analysis of Srassica /uncea * Buffalo dung (mature)

Caption: Graph No. : 4.7 FT- IR analysis of Srassica /uncea * Eisenia toot'd a ? Buffalo dung
Table 1. Effect of treatments on yield of
vermicompost (kg)

Treatments        Yield (kg)

Control            1.60 (a)
[T.sub.1]          2.60 (b)
[T.sub.2]          2.25 (a,b)
[T.sub.3]          3.80 (c)
SEM ([+ or -])     0.33
CD 5%              0.75

Control: Buffalo dung + Eisenia foetida; [T.sub.1]:
Buffalo dung + Brassica juncea cake; [T.sub.2]:
Buffalo dung + Brassica juncea cake + Eisenia
foetida; and [T.sub.3]: Brassica juncea cake +
Eisenia foetida

Alphabet (a,b,c) shows significance and
nonsignificance of treatments at 5% level of
significance. Treatments with same alphabet
are non-significant and with different alphabet
aresignificant.

Table 2. Effect of treatments on pH, EC, Total organic carbon (TOC)
contents at various stages (0, 30, 60 and 90 days) of vermicomposting

                                       pH

Treatments          0            30        60        90

Control          6.25 (a,b)   6.64 (a,b)   6.90    7.46 (a)
[T.sub.1]        6.13 (a)     6.33 (a)     6.76    7.43 (a)
[T.sub.2]        6.33 (a,b)   6.53 (a,b)   6.73    7.83 (a,b)
[T.sub.3]        6.56 (b)     6.78 (b)     7.10    7.96 (b)
SEM([+ or -])    0.14         0.14         0.16    0.19
CD5%             0.31         0.31          NS     0.45

                      EC (dS[m.sup.-1])

Treatments       0      30       60       90

Control         0.21   0.21   0.23 (a)   0.37
[T.sub.1]       0.27   0.31   0.38 (b)   0.47
[T.sub.2]       0.26   0.28   0.33 (b)   0.46
[T.sub.3]       0.21   0.31   0.37 (b)   0.41
SEM([+ or -])   0.03   0.05   0.04       0.05
CD5%             NS     NS    0.09        NS

                           TOC (%)

Treatments        0      30      60      90

Control         14.72   13.99   13.25   13.11
[T.sub.1]       15.24   14.36   14.01   13.95
[T.sub.2]       15.31   14.99   13.97   12.41
[T.sub.3]       14.98   14.44   13.30   12.15
SEM([+ or -])    1.82    1.93    2.57    1.69
CD5%              NS      NS      NS      NS

Control: Buffalo dung + Eisenia foetida; [T.sub.1]: Buffalo
dung + Brassica juncea cake;[T.sub.2]: Buffalo dung + Brassica
juncea cake + Eisenia foetida; and [T.sub.3]: Brassica juncea
cake + Eisenia foetida

Alphabet (a,b) shows significance and non-significance of
treatments at 5% level of significance. Treatments with
same alphabet are non-significant and with different
alphabet are significant.

Table 3. Changes in available phosphorus, available
potassium and available sodium at various stages
(0, 30, 60 and 90 days) of vermicomposting

                             Available Phosphorus (%)

Treatments            0             30            60          90

Control           0.023 (a)      0.028 (a)     0.033 (a)   0.041 (a)
[T.sub.1]         0.025 (a)      0.029 (a)     0.034 (a)   0.041 (a)
[T.sub.2]         0.033 (a,b)    0.038 (a,b)   0.048 (b)   0.061 (b)
[T.sub.3]         0.044 (b)      0.051 (b)     0.061 (c)   0.074 (c)
SEM([+ or -])     0.005          0.006         0.003       0.004
CD5%              0.012          0.014         0.007       0.009

                    Available Potassium (%)

Treatments         0      30      60      90

Control          0.053   0.075   0.085   0.099
[T.sub.1]        0.054   0.066   0.079   0.098
[T.sub.2]        0.057   0.072   0.086   0.100
[T.sub.3]        0.050   0.074   0.089   0.096
SEM([+ or -])    0.007   0.008   0.006   0.007
CD5%              NS      NS      NS      NS

                        Available Sodium (%)

Treatments         0      30         60           90

Control          0.024   0.032   0.036 (a,b)   0.041 (a)
[T.sub.1]        0.022   0.027   0.031 (a)     0.039 (a)
[T.sub.2]        0.030   0.035   0.043 (b)     0.059 (b)
[T.sub.3]        0.021   0.026   0.032 (a,b)   0.040 (a)
SEM([+ or -])    0.005   0.004   0.005         0.005
CD5%              NS      NS     0.011         0.011

Control: Buffalo dung + Eisenia foetida; [T.sub.1]: Buffalo
dung + Brassica juncea cake; [T.sub.2]: Buffalo dung + Brassica
juncea cake + Eisenia foetida; and [T.sub.3]: Brassica juncea
cake + Eisenia foetida

Alphabet (a,b,c) shows significance and non-significance of
treatments at 5% level of significance. Treatments with
same alphabet are non-significant and with different
alphabet are significant.

Table 4. Changes in lignin, cellulose and hemicellulose
at various stages (0, 30, 60 and 90 days) of vermicomposting

                                    Lignin (%)

Treatments            0          30           60          90

Control            2.71 (a)    3.12 (a)    3.56 (a)     4.25 (a)
[T.sub.1]         16.32 (b)   20.25 (b)   24.36 (b)    31.25 (b)
[T.sub.2]         17.74 (c)   22.13 (c)   25.70 (c)    31.25 (b)
[T.sub.3]         30.02 (d)   36.25 (d)   40.39 (d)    48.36 (c)
SEM ([+ or -])    0.39         0.54        0.27         0.74
CD5%              0.91         1.25        0.61         1.71

                                  Cellulose ([micro]g)

Treatments            0           30          60          90

Control           26.24 (a)    19.71 (a)   16.01 (a)   14.65 (a)
[T.sub.1]         25.60 (a)    22.27 (b)   17.95 (b)   15.74 (b)
[T.sub.2]         31.68 (b)    23.29 (c)   19.55 (c)   14.04 (a)
[T.sub.3]         33.48 (c)    28.29 (d)   18.30 (b)   12.09 (c)
SEM ([+ or -])     0.65         0.30        0.41        0.45
CD5%               1.50         0.68        0.94        1.04

                                Hemicellulose (%)

Treatments            0          30          60          90

Control            2.71 (a)    2.10 (a)    1.71 (a)    1.49 (a)
[T.sub.1]         16.99 (b)   16.12 (b)   15.26 (b)   15.02 (b)
[T.sub.2]         15.54 (c)   13.69 (c)   11.25 (c)    9.64 (c)
[T.sub.3]         29.99 (d)   25.63 (d)   26.25 (d)   22.36 (d)
SEM ([+ or -])     0.38        0.42        0.33        0.45
CD5%               0.88        0.96        0.76        1.03

Control: Buffalo dung + Eisenia foetida; [T.sub.1]: Buffalo
dung + Brassica juncea cake; [T.sub.2]: Buffalo dung + Brassica
juncea cake + Eisenia foetida; and [T.sub.3]: Brassica juncea
cake + Eisenia foetida

Alphabet (a,b,c,d) shows significance and non-significance of
treatments at 5% level of significance. Treatments with same
alphabet are non-significant and with different alphabet
are significant

Table 5. Specific functional group/compound present in control
of raw buffalo dung and dung composted with Eisenia foetida

Raw buffalo dung

Adsorption          Functional group
frequency           or compound
([cm.sup.-1])

3420.85             Moisture
3011.51             Aromatic C-H structure
2933.28             C-H with methyl and
                    methylene
2226.58             C[O.sub.2]
1487.17             Aromatic monomer
1423.99             C-N stretching/-OH bond
                    of carboxyl
1296.03             Phosphate ester
1052.37             Ethanol
848.80              Aromatic carbon compound

Raw buffalo dung    Dung composted with Eisenia foetida

Adsorption          Adsorption      Functional group
frequency           frequency       or compound
([cm.sup.-1])       ([cm.sup.-1])

3420.85             3397.69         Heteroaromatic compound
3011.51                             containing N-H group
2933.28             2352.87         C[O.sub.2]
                    1049.32         C-O starching vibration in
2226.58                             alcohol and phenol
1487.17             829.79          Aromatic compound region
1423.99

1296.03
1052.37
848.80

Table 6. Specific functional group /compound present in control of
Brassica juncea and Brassica juncea composted with Eisenia foetida

Raw Brassica juncea

Adsorption       Functional group
frequency        or compound
([cm.sup.-1])

3421.91          Two N-H group
                 (primary amine and amide)
3009             Heteroaromatic like pyridine,
                 parazine pyrols, furan
2924.11          C-H with methyl and methylene
2352.96          C[O.sub.2]
1637.94          C-O,C-N,N-O
1425.55          C-N stretching
1287.52          Aromatic/ heteromatic carbon
848.80           Aromatic carbon compound

Brassica juncea composted with Eisenia foetida

Adsorption       Functional group
frequency        or compound
([cm.sup.-1])

3396.59          Heteroaromatic compound containing
                 N-H group
3007.63          Heteroaromatic like pyridine,
                 parazine pyrols, furan
2927.48          Asymmetric/symmetric stretching
                 of methylene
2208.24          Weak (C=N) stretching bond alkaline
                 molecule, C=C.
1420.35          Aromatic/ heteroaromatic ,
                 C-N stretching
1036.22          C-O bond, C-O stretching vibrations
                 in alcohol and phenol

Table 7. Specific functional group/compound present in control
of Brassica juncea+dung and Brassica juncea composted with dung

Control of Brassica juncea + dung

Adsorption       Functional group
frequency        or compound
([cm.sup.-1])

3410.48          Two N-H group(primary amine
                 and amide)
3006.49          Heteroaromatic like pyridine,
                 parazine pyrols, furan
2924.32          Asymmetric/symmetric
                 stretching of methylene
1937.33          C=C-C[H.sub.2]
1637.09          C-O,C-N,N-O
1425.48          C-N stretching
1488.76          Aromatic/ heteroaromatic compound
848.80           Aromatic carbon compound

Composted Brassica juncea +dung

Adsorption       Functional group
frequency        or compound
([cm.sup.-1])

3404.52          Heteroaromatic compound containing
                 N-H group, Two N-H group(primary
                 amine and amide)
2925.54          Asymmetric/symmetric stretching
                 of methylene
1054.53          Quinones
3002.60          Aromatic C-H stretching
1420, 1485.66    Aromatic/ heteroaromatic, C-N stretching
1296             C-O,C-N, phosphate ester
874.38,          Interaction between C=S and N,
888.58,          Si-H Bend.
841.33

Table 8. Specific functional group /compound present in control
of dung+ Brassica juncea and Brassica juncea composted
with Eisenia foetida +dung

Control of Brassica juncea + dung

Adsorption       Functional group
frequency        or compound
([cm.sup.-1])

3410.48          Two N-H group(primary amine
                 and amide)
3006.49          Heteroaromatic like pyridine,
                 parazine pyrols, furan
2924.32          Asymmetric/symmetric
                 stretching of methylene
1937.33          C=C-C[H.sub.2]
1637.09          C-O,C-N,N-O
1425.48          C-N stretching
1488.76          Aromatic/ heteroaromatic compound
848.80           Aromatic carbon compound

Composted Brassica juncea +dung+ Eisenia foetida

Adsorption       Functional group
frequency        or compound
([cm.sup.-1])

3401.85          Two N-H group(primary amine and
                 amide) Heteroaromatic compound
                 containing N-H group
2925.09          Asymmetric/symmetric stretching of
                 methylene.
2997.91          C-H with methyl and methylene
2085.70          C=C,C=N
1654.56          Quinone
1420, 1485.7     Aromatic / heteroaromatic C-N stretching.
1297             C-C,C-O,C-N, phosphate ester
1037             C-O stretching vibration in alcohol a
                 and phenol
832              C-C,C-O,C-N, furan ring
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Author:Solanki, Vikram C.; Shilpkar, Prateek
Publication:Journal of Pure and Applied Microbiology
Article Type:Report
Date:Jun 1, 2016
Words:4385
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