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Bio degradation of organic wastes by Pleurotus eous mushroom

Authors:
  • Adhiparasakthi Agricultural College

Abstract and Figures

Composting is an important strategy for the management of organic wastes to get quality end product with better microbial community. Various fungi and bacteria including mushrooms were selected for degradation of agro wastes. The studies were conducted on the following aspects viz., collection and utilization of locally available agricultural residues as substrates, addition of inorganic and organic additives to substrates along with Pleurotus eous (APK1) mushroom for decomposition. The substrates like sugarcane trash, sugarcane bagasse, groundnut shell, lawn grass and coir waste and additives such as urea, gypsum, calcium carbonate, calcium hydroxide, groundnut oil cake, gingelly oil cake and neem oil cake were used in this study. The components viz., carbon, nitrogen and crude fibre with each additive were analysed after incubation period. Sugarcane trash with urea decomposed highly with minimum C: N ratio. Crude fibre content was less in groundnut shell with urea.
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~ 1542 ~
Journal of Pharmacognosy and Phytochemistry 2020; 9(3): 1542-1545
E-ISSN: 2278-4136
P-ISSN: 2349-8234
www.phytojournal.com
JPP 2020; 9(3): 1542-1545
Received: 20-03-2020
Accepted: 24-04-2020
S Karpagavalli
Associate Professor, Department
of Plant Pathology,
Adhiparasakthi Agricultural
College G.B. Nagar, Kalavai,
Tamil Nadu, India
K Ganeshkumar
Research Students, Department
of Plant Pathology,
Adhiparasakthi Agricultural
College G.B. Nagar, Kalavai,
Tamil Nadu, India
K Jayaprakash
Research Students, Department
of Plant Pathology,
Adhiparasakthi Agricultural
College G.B. Nagar, Kalavai,
Tamil Nadu, India
Corresponding Author:
S Karpagavalli
Associate Professor, Department
of Plant Pathology,
Adhiparasakthi Agricultural
College G.B. Nagar, Kalavai,
Tamil Nadu, India
Bio degradation of organic wastes by Pleurotus
eous mushroom
S Karpagavalli, K Ganeshkumar and K Jayaprakash
Abstract
Composting is an important strategy for the management of organic wastes to get quality end product
with better microbial community. Various fungi and bacteria including mushrooms were selected for
degradation of agro wastes. The studies were conducted on the following aspects viz., collection and
utilization of locally available agricultural residues as substrates, addition of inorganic and organic
additives to substrates along with Pleurotus eous (APK1) mushroom for decomposition. The substrates
like sugarcane trash, sugarcane bagasse, groundnut shell, lawn grass and coir waste and additives such as
urea, gypsum, calcium carbonate, calcium hydroxide, groundnut oil cake, gingelly oil cake and neem oil
cake were used in this study. The components viz., carbon, nitrogen and crude fibre with each additive
were analysed after incubation period. Sugarcane trash with urea decomposed highly with minimum C: N
ratio. Crude fibre content was less in groundnut shell with urea.
Keywords: Pleurotus eous, agricultural residues (substrates), organic additives, inorganic additives,
carbon, nitrogen and crude fibre
Introduction
Crop residues are the noneconomic plant parts that are left in the field after harvest. The
harvested refuses include straws, stubble, stover and hulms of different crops. Crop remains
are also from thrashing sheds or that are discarded during crop processing. This includes
wastes like groundnut shell, oil cakes, rice husks and cobs of maize, sorghum and cumbu.
India’s total amount of agro-industrial residues reaches 600 million tonnes, they were crop
based residues (generated in field) and processing based residues (generated during wood and
industrial processing) (Mande, 2005) [4]. The greatest potential as a biomass resource appears
to be from the field residues of sorghum, maize, soyabean, cotton, sugarcane etc. these
residues need composting before being used as manure. Compost provides a stable organic
matter that enhance the soil quality and crop production. Compost application improved the
soil properties by creating suitable conditions for root development and consequently
promoting higher yield and higher quality of crops. Composting is brought about by a variety
of organisms like bacteria, fungi and actinomycetes. Fungi grow in nature on all sorts of
vegetable wastes and they play an important part in maintaining the natural cycle. Among the
fungi mushrooms are capable of breaking down organic material than other microorganisms.
Composting provides a medium favourable for the mushroom spawn to predominate. With this
idea the research work was carried out by using Pleurotus eous (APK 1) as biodegrading agent
for decomposition of agricultural wastes (substrates) along with organic and inorganic
additives for the estimation of carbon, nitrogen and crude fibre.
Materials and Methods
Preparation of mother spawn and bed spawn: spores of Pleurotus eous (APK 1) were
collected directly from the fruiting bodies and inoculated on Potato Dextrose Agar (PDA)
medium. The inoculated Petri plates were incubated at 15 oC for three days. The mycelium of
P. eous appeared on the Petri plates were used as inoculum. Half cooked sorghum grains were
mixed with the calcium carbonate (CaCO3) @ 20g/ kg. Calcium carbonate was added to
absorbs excess moisture and neutralize the pH. The prepared grains were filled in to
polypropylene bags and sterilized at 15 lbs pressure for one hour. The mycelium of P. eous
appeared in the Petri plate was inoculated into sterilized cooled spawn bags. Mouth of the bags
were close with non-absorbent cotton plug and then incubated at 22 24 oC in a dark place.
The mycelium completely spread through the grains in about two weeks from this fully grown
mother spawn bags, bed spawns were prepared by inoculating few grains with mycelium of P.
eous to sterilized cooled sorghum grains in polypropylene bags and incubated as described
earlier. Bed spawn bags were used for decomposition.
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Journal of Pharmacognosy and Phytochemistry http://www.phytojournal.com
Substrates used for decomposition
The following substrates were collected from Adhiparasakthi
Agricultural College campus and in and around G.B. Nagar,
Kalavai.
1. Sugarcane trash
2. Sugarcane bagasse
3. Groundnut shell
4. Coir waste and
5. Lawn grass (Zoisia tenuifolia)
Additives used for decomposition
1. Urea
2. Gypsum
3. Calcium carbonate
4. Calcium hydroxide
5. Gingelly oilcake
6. Groundnut oilcake
7. Neem oilcake
Urea and gypsum purchased from fertilizer shop, calcium
carbonate and calcium hydroxide from scientific companies
and oil cakes were also obtained from local market of
Kalavai.
Preparation of beds for decomposition
The substrates and additives were added layer by layer in
polythene bags (60 x30 cm) with 40 per cent moisture
content.
In each bag one kg of substrates was added in five layers
along with 2g of additive per layer. Pleurotus eous added in
each layer at the rate of 10g and the bags were tied and
incubated at room temperature. Few holes were made in bags
to provide aeration. Three replications were maintained for
each substrate with each additive. Samples are taken from
each bag on 75 days after incubation. Dried the substrates and
powdered and then used for estimation of C: N ratio and
crude fibre.
Treatment details for each bag
1. Sugarcane trash
Sugarcane trash + Pleurotus eous spawn + urea
Sugarcane trash + Pleurotus eous spawn + gypsum
Sugarcane trash + Pleurotus eous spawn + calcium carbonate
Sugarcane trash + Pleurotus eous spawn + calcium hydroxide
Sugarcane trash + Pleurotus eous spawn + ground nut oil
cake
Sugarcane trash + Pleurotus eous spawn + gingelly oil cake
Sugarcane trash + Pleurotus eous spawn + neem oil cake
2. Sugarcane bagasse
Sugarcane bagasse + Pleurotus eous spawn + urea
Sugarcane bagasse + Pleurotus eous spawn + gypsum
Sugarcane bagasse + Pleurotus eous spawn + calcium
carbonate
Sugarcane bagasse + Pleurotus eous spawn + calcium
hydroxide
Sugarcane bagasse + Pleurotus eous spawn + ground nut oil
cake
Sugarcane bagasse + Pleurotus eous spawn + gingelly oil
cake
Sugarcane bagas se+ Pleurotus eous spawn + neem oil cake
3. Groundnut shell
Groundnut shell+ Pleurotus eous spawn + urea
Groundnut shell + Pleurotus eous spawn + gypsum
Groundnut shell + Pleurotus eous spawn + calcium carbonate
Groundnut shell + Pleurotus eous spawn + calcium hydroxide
Groundnut shell + Pleurotus eous spawn + ground nut oil
cake
Groundnut shell + Pleurotus eous spawn + gingelly oil cake
Groundnut shell + Pleurotus eous spawn + neem oil cake
4. Coir waste
Coir waste + Pleurotus eous spawn + urea
Coir waste + Pleurotus eous spawn + gypsum
Coir waste + Pleurotus eous spawn + calcium carbonate
Coir waste + Pleurotus eous spawn + calcium hydroxide
Coir waste + Pleurotus eous spawn + ground nut oil cake
Coir waste + Pleurotus eous spawn + gingelly oil cake
Coir waste + Pleurotus eous spawn + neem oil cake
5. Lawn grass
Lawn grass + Pleurotus eous spawn + urea
Lawn grass + Pleurotus eous spawn + gypsum
Lawn grass + Pleurotus eous spawn + calcium carbonate
Lawn grass + Pleurotus eous spawn + calcium hydroxide
Lawn grass + Pleurotus eous spawn + ground nut oil cake
Lawn grass + Pleurotus eous spawn + gingelly oil cake
Lawn grass + Pleurotus eous spawn + neem oil cake
Carbon estimation (Walkley and Black, 1934) [7].
The following reagents viz., 1 N potassium dichromate,
concentrated sulphuric acid, 85 per cent orthophosphoric acid,
phenylamine indicator and 0.5 N ferrous ammonium sulphate
were used for carbon estimation.
Organic carbon content of the sample was estimated as per the
method described by Walkley and Black (1934) [7]. Hundred
mg of the sample was added with 20 ml of 1 N potassium
dichromate and 20 ml of concentrated sulphuric acid. The
content was digested for 30 min. a blank was also run
simultaneously. Then 250 ml of distilled water followed by 1
ml of 85 per cent ortho phosphoric acid and 0.5 ml of
diphenylamine indicator were added. The content of the flask
was back titrated with 0.5 N ferrous ammonium sulphate
solution. The titration was continued until the appearance of a
brilliant green colour, indicating the end point. The organic
content was expressed in percentage using the formula.
Calculation
Where
BV, is blank value
SV, sample value and
W, weight of the sample
Nitrogen estimation (Amma,1989)
The following reagents viz., concentrated sulphuric acid, Salt
mixture of potassium sulphate (25g), copper sulphate (5g) and
metallic selenium (0.5g), boric acid 2 per cent containing
bromocresol green methyl red indicator, sodium hydroxide 40
per cent solution, 0.1 N sodium carbonate and 0.002 N
hydrochloric acid.
Digestion
Transferred 250 mg of dried plant sample in to 100 ml
Kjeldhal flask and added with 200 mg of salt mixture
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Journal of Pharmacognosy and Phytochemistry http://www.phytojournal.com
(digestion mixture), 10 g of potassium sulphate + 1 g copper
sulphate (10:1). Then added 3 ml of concentrated sulphuric
acid (let the content react for I hour). Transferred the Kjeldhal
flask to digestion unit. Transferred the heat briskly for
subsequent digestion and continued the heating until the
digest become clear (colourless digestion mixture will be
obtained). Cooled the Kjeldhal flask.
Distillation
Transferred the whole plant digest material to distillation
assembly. Rinsed the Kjeldhal flask thrice with 5 ml of
distilled water. Added 10 ml of 40 per cent sodium hydroxide.
Heated the flask filled with redistilled water to boil and the
boiling steam was passed in to distillation flask. Kept a 100
ml beaker containing 20 ml of 2 per cent boric acid with
double indicator below the delivery end of the condenser in
the distillation set. Distilled the content and collected the
liberated ammonia in boric acid. Continued the distillation
until release free of ammonia about 30 ml distillate was
collected (tested by litmus paper near the out let of the
condenser which was turn blue as ammonia was present).
Titrated the ammonia collected boric acid with N/10 sulphuric
acid. End point was appearance of wine red colour.
Calculation
Where
S, is ml of standard acid used in the titration of the sample
distillate
B, is the ml of standard acid used in the titration of the blank
distillate
N, is the normality of the acid used in the titration of the
distillate
ME, is the milligram equivalent weight of the nitrogen i.e.
0.014
W, is the mass of plant sample taken in gram for the
preparation of the sample digest.
Estimation of crude fibre
Procedure
Extracted 2g of ground material with ether or petroleum ether
to remove fat (initial boiling temperature 35 38 oC and final
temperature 52 oC) if fat content is below 1 per cent extraction
may be omitted. After extraction with ether boiled 2g of dried
material with 200ml of sulphuric for 30min. with bumping
chips. Filtered through muslin cloth and washed with boiling
water until washing were no longer acidic. Boiled with 200ml
of sodium hydroxide solution for 30min. Filtered through
muslin cloth again and washed with 25ml of boiling 1.25 per
cent sulphuric acid, three portions 50 ml of water and 25 ml
of alcohol. Removed the residue and transferred to ashing
dish (pre weighed dish w1). Dried the residue for 2 hours at
130 ± 2 oC. Cooled the dish in a desicator and weighed (w2).
Ignited for 30 min. at 600 ± 15 oC. Cooled in a desiccator and
reweighed (w3).
Calculation
Percentage of crude fibre in ground sample
Statistical analysis
Data of the experiments were analysed by Factorial
Completely Randamized Block Design (CRD) using data
entry module for Ag Res Statistical Software© 1994 Pascal
International Software Solutions, version 3.01 for data entry
and version 7.01 for analysis.
Results and Discussion
Organic manures are highly regarded as good source of
material to maintain soil health and increasing soil organic
carbon content. Various agro-wastes are converted in to
organic manure by composting in which biodegradation with
microorganisms especially mushrooms are highly used.
Species of oyster mushrooms are able to degrade cellulose,
lignin and hemicellulose through their enzyme production. In
this study, the oyster mushroom, Pleurotus eous (APK 1) was
used to degrade various agro-wastes (substrates). Dried and
powdered samples of substrates were used from each bag for
the estimation of C: N ratio and crude fibre.
Estimation of C: N ratio on biodegradation of various
agro-wastes by Pleurotus eous mushroom
Experiment was conducted to study C: N ratio of agro- wastes
with additives on biodegradation by P. eous and results are
summarized in the Table 1.
Carbon: Nitrogen ratio was estimated in all substrates with
each additives on 75 days after incubation. Among the
substrates sugarcane trash with urea additive was decomposed
much with C: N ratio of 23. 08: 1 followed by lawn grass
25.10: 1 and sugar cane bagasse 25.90: 1.
Table 1: Estimation of C:N ratio on biodegradation of various agro-wastes by Pleurotus eous mushroom
S. No.
Additives
Carbon: Nitrogen ratio (%)
Substrates
Sugarcane trash
Groundnut shell
Lawn grass
Coir waste
1.
Urea
23.80: 1a
29.20: 1 f
25.10: 1 b
32.30: 1 h
2.
Gypsum
27.40: 1d
32.20: 1 h
29.20: 1 f
36.40: 1 k
3.
Calcium carbonate
30.30: 1 g
36.13: 1 j
33.20: 1 i
40.20: 1 n
4.
Calcium hydroxide
33.26: 1 i
39.30: 1 m
38.20: 1 l
44.31: 1 p
5.
Groundnut oilcake
39.40: 1 m
44.20: 1 p
44.35: 1 p
49.26: 1 t
6.
Gingelly oil cake
42.67: 1 o
48.12: 1 s
50.41: 1 v
55.53: 1 w
7.
Neem oil cake
46.32: 1 q
56.45: 1 x
57.21: 1 y
61.23: 1 z
8.
Control
83.30: 1 €
75.28: 1 |
78.10: 1 }
81.08: 1~
S T ST Note: { > | >} > ~ > €
SED 0.02962 0.03746 0.08377
CD (0.05) 0.05894 0.07456 0.16672
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Journal of Pharmacognosy and Phytochemistry http://www.phytojournal.com
On biodegradation, coir waste showed minimum ratio of C: N
that indicates the reduced bio-efficiency. Within the additives
urea was effective in degradation of all substrates followed by
gypsum. All other chemical and natural additives showed less
degradation.
This result was coincided with the report of Pandey et al.
(2012) [5]. They reported that sugar bagasse treated with urea
and hot water along with Pleurotus citrinopileatus decreased
the carbon content and increased the nitrogen content at
different stages of growth. It showed the activities of
cellulolytic, hemi cellulolytic and lignolytic enzymes
produced by Pleurotus citrinopileatus. Theradimani et al.
(2018) [6] studied that the inoculation of coir waste with six
Pleurotus spp., antagonists and biofertilizers reduced the
organic carbon content. Maximum reduction in organic
carbon content of coconut coir pith (37.53 per cent) and
maximum increase in total nitrogen (318.5 per cent) was
brought about by the inoculation of Pleurotus djamor. In this
experiment, the C:N ratio also reduced in coconut coir pith as
32.30:1 by Pleurotus eous after 75 days of incubation as
compared to 81.08: 1 in control. Comparison of all substrates,
sugarcane trash with urea additive was decomposed much by
P. eous.
Estimation of crude fibre on biodegradation of various
agro-wastes by Pleurotus eous mushroom
Experiment was conducted to study crude fibre of agro-wastes
with additives on biodegradation by P. eous and results are
summarized in the Table 2.
Crude fibre was an insoluble residue that remains after
decomposition of plant materials. In this experiment
biodegradation with P.eous revealed the content of crude fibre
after 75 days of incubation (decomposition) in various
substrates with additives. It indirectly showed the
decomposition percentage of substrates with the influence of
both inorganic and organic additives. Among the substrates
used, groundnut shell with urea showed the minimum crude
fibre content of 1.60 per cent followed by groundnut shell
with gypsum as 1.80 per cent. Within the substrates
groundnut shell with all additives showed less crude fibre
followed by sugarcane trash and lawn grass. Maximum crude
fibre was obtained from coir waste and sugarcane bagasse. Oil
cakes of groundnut, gingelly and neem showed more fibre in
all substrates due to less decomposition.
The result was corroborated with the report of El-Mandany
(1997) [3] as crude fibre content decreased from 33.64 and
32.29 per cent for urea treated rice straw. Adenipekun and
Dada (2013) [`1] also reported that crude fibre decreased
significantly in cotton waste and cocoa husk from 5.88 to 5.31
per cent and from 39.88 to 34.95 per cent respectively
whereas it was increased in rice straw from 18.42 to 28.08 per
cent after 60 days of incubation with Pleurotus pulmonarius.
In this experiment also groundnut shell with urea followed by
groundnut shell with gypsum showed the minimum crude
fibre content. Higher content of crude fibre was obtained from
coir waste and sugarcane bagasse. Oil cakes of groundnut,
gingelly and neem showed more fibre in all substrates due to
less decomposition.
Table 2: Estimation of crude fibre on biodegradation of various agro-wastes by Pleurotus eous mushroom
S. No.
Additives
Crude fibre (%)
Substrates
Sugarcane trash
Sugarcane bagasse
Groundnut shell
Lawn grass
Coir waste
1.
Urea
20.12 i
23.20 l
1.60 a
21.32 j
22.51 k
2.
Gypsum
22.35 k
25.60 p
1.80 b
23.50 m
24.53 o
3.
Calcium carbonate
24.33 n
27.25 r
2.10 c
26.20 q
27.21 r
4.
Calcium hydroxide
25.43 p
29.45 v
2.50 d
28.80 t
29.40 u
5.
Groundnut oilcake
28.45 s
32.61 {
3.21 e
30.33 v
32.63 z
6.
Gingelly oil cake
30.37 v
34.50 ~
4.16 f
31.60 x
34.05}
7.
Neem oil cake
31.23 w
36.28 □
4.79 g
32.65 y
35.30
8.
Control
33.50 |
38.17 □
7.50 h
34.10 }
36.24 €
S T ST Note: { >l >} > ~ > □ > €
SED 0.02877 0.03639 0.08136
CD (0.05) 0.05725 0.07241 0.16192
Conclusion
Pleurotus eous decomposed all the substrates used in the
study in which sugarcane trash with urea was highly
decomposed with minimum C: N ratio. Crude fibre content
was less in groundnut shell with urea.
Reference
1. Adenipekun CO, Dada OJ. Biodegradation of three
Agricultural Wastes by a white rot fungus Pleurotus
pulmonarius (Fries) Quetlet. Nature and Science. 2013;
11(2):19-25.
2. Amma T. Nitrogen estimation in plant sample. Nitrogen
determination by micro-kjeldhal digestion and distillation
method, 1989, 261-264.
3. El-Mandany HA. Nutritional using of treated rice straw
in goat rations. M.Sc. Thesis. Fac. Of Agric., Zagazig
University, 1997.
4. Mande S. Biomass gasifier-based power plants: potential,
problems and research needs for decentralized rural
electrification. In: Lal B, Reddy MRVP (eds) Wealth
from Waste: Trends and Technologies, New Delhi, 2005.
5. Pandey A, Soccol CR, Mitchell D. New developments in
solid state fermentation: I- Bioprocesses and products.
Process. Biochem. 2012; 35:1153-1169.
6. Theradimani M, Thangeshwari S, Parthasarathy S.
Biological decomposition of coconut coir pith waste.
Pl.Dis. Res. 2018; 33(2):142-147.
7. Walkley A, Black CA. An examination of Degtjareff
method by determining soil organic matter and a
proposed modification of the chromic acid titration
method. Soil Science. 1934; 37:29-38.
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Article
Full-text available
The effect of different white rot fungi including Pleurotus djamor, Pleurotus eous, Pleurotus sajor-caju, Pleurotus florida and other basidiomycetes fungi, Calocybe indica, Hypsizygus ulmarius, antagonist’s organism viz., Trichoderma viride, Pseudomonas fluorescens, nitrogen fixing bacteria viz., Azospirillum and Phosphobacteria on coconut coirpith decomposition was studied. The coirpith inoculated with different organisms reduced organic carbon content and increased total nitrogen content significantly. However, the maximum reduction of C: N ratio, cellulose and lignin was observed in P. djamor, P. sajor-caju and T. viride. The decomposed coirpith had increased nitrogen, phosphorus, potash and calcium content.
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Nitrogen estimation in plant sample. Nitrogen determination by micro-kjeldhal digestion and distillation method
  • T Amma
Amma T. Nitrogen estimation in plant sample. Nitrogen determination by micro-kjeldhal digestion and distillation method, 1989, 261-264.
Nutritional using of treated rice straw in goat rations
  • H A El-Mandany
El-Mandany HA. Nutritional using of treated rice straw in goat rations. M.Sc. Thesis. Fac. Of Agric., Zagazig University, 1997.
Biomass gasifier-based power plants: potential, problems and research needs for decentralized rural electrification
  • S Mande
Mande S. Biomass gasifier-based power plants: potential, problems and research needs for decentralized rural electrification. In: Lal B, Reddy MRVP (eds) Wealth from Waste: Trends and Technologies, New Delhi, 2005.