An Outlook of Vermicomposting and its Scope in Future.pdf
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JCBPS; Section D; November 2016 – January 2017, Vol. 7, No. 1; 049-065. E- ISSN: 2249 –1929
Journal of Chemical, Biological and Physical Sciences
An International Peer Review E-3 Journal of Sciences
Available online atwww.jcbsc.org
Section D: Environmental Sciences
CODEN (USA): JCBPAT Review Article
49 J. Chem. Bio. Phy. Sci. Sec. D, November 2016 – January 2017; Vol.7, No.1; 049-065.
An Outlook of Vermicomposting and its Scope in
Future
1*
Abul Kalam and
2
Syed Rehan Ahmad
1
Department of Microbiology, Bidhannagar Govt College, Salt Lake City, Kolkata, India
2
Department of Microbiology, Shri Venkateshwara University, Gajraula, U.P, India.
Received: 27 October 2016; Revised: 01 December 2016; Accepted: 07 December 2016
Abstract: Vermicompost is the item or procedure of fertilizing the soil utilizing
different worms, typically red wigglers, white worms, and different night crawlers, to
make a heterogeneous blend of disintegrating vegetable or nourishment squander,
bedding materials, and vermicast, additionally called worm castings, worm humus or
worm fertilizer, is the finished result of the breakdown of natural matter by an
earthworm. These castings have been appeared to contain lessened levels of
contaminants and a higher immersion of supplements than do natural materials before
vermicomposting. Containing water-dissolvable supplements, vermicompost is an
amazing, supplement rich natural compost and soil conditioner. This procedure of
delivering vermicompost is called vermicomposting. The present article deal with An
Outlook of Vermicomposting and its scope in future.
Keywords: Vermicompost, Bioteks, Biotechnology
INTRODUCTION
Vermicompost is the end-product of the breakdown of organic matter by some species of earthworm.
Vermicompost is a nutrient-rich, organic fertilizer and soil conditioner. The process of producing
vermicompost is called vermicomposting. It is the excreta of earthworm, which is rich in humus.
Earthworms eat cow dung or farm yard manure along with other farm wastes and pass it through their
body and in the process convert it into vermicomposting,
So Vermicompost = worm castings + humus + microorganisms + partially decomposed bedding
and plant materials.]
Journal of Chemical, Biological and Physical Sciences
An International Peer Review E-3 Journal of Sciences
Available online atwww.jcbsc.org
Section D: Environmental Sciences
CODEN (USA): JCBPAT Review Article
49 J. Chem. Bio. Phy. Sci. Sec. D, November 2016 – January 2017; Vol.7, No.1; 049-065.
An Outlook of Vermicomposting and its Scope in
Future
1*
Abul Kalam and
2
Syed Rehan Ahmad
1
Department of Microbiology, Bidhannagar Govt College, Salt Lake City, Kolkata, India
2
Department of Microbiology, Shri Venkateshwara University, Gajraula, U.P, India.
Received: 27 October 2016; Revised: 01 December 2016; Accepted: 07 December 2016
Abstract: Vermicompost is the item or procedure of fertilizing the soil utilizing
different worms, typically red wigglers, white worms, and different night crawlers, to
make a heterogeneous blend of disintegrating vegetable or nourishment squander,
bedding materials, and vermicast, additionally called worm castings, worm humus or
worm fertilizer, is the finished result of the breakdown of natural matter by an
earthworm. These castings have been appeared to contain lessened levels of
contaminants and a higher immersion of supplements than do natural materials before
vermicomposting. Containing water-dissolvable supplements, vermicompost is an
amazing, supplement rich natural compost and soil conditioner. This procedure of
delivering vermicompost is called vermicomposting. The present article deal with An
Outlook of Vermicomposting and its scope in future.
Keywords: Vermicompost, Bioteks, Biotechnology
INTRODUCTION
Vermicompost is the end-product of the breakdown of organic matter by some species of earthworm.
Vermicompost is a nutrient-rich, organic fertilizer and soil conditioner. The process of producing
vermicompost is called vermicomposting. It is the excreta of earthworm, which is rich in humus.
Earthworms eat cow dung or farm yard manure along with other farm wastes and pass it through their
body and in the process convert it into vermicomposting,
So Vermicompost = worm castings + humus + microorganisms + partially decomposed bedding
and plant materials.]
An Outlook … Kalam and Ahmad.
50 J. Chem. Bio. Phy. Sci. Sec. D, November 2016 – January 2017; Vol.7, No.1; 049-065.
Vermiculture has been the primary focus at Maharashtra Agricultural Bioteks in India, an
organization which has initiated both commercial and educational ventures to promote Vermiculture.
In 1985, Maharashtra Agricultural Bioteks was formed and established a small plant to manufacture
vermicompost from agricultural waste. In 1991-92, Maharashtra Bioteks and the India Department of
Science and Technology promoted the adoption of vermicompost technology in 13 states in India. The
group has also established a vermicompost unit with Chitrakoot Gramodaya University, Madhya
Pradesh which produces five tons of vermicompost per month.
Advantages of vermicompost:
1. Vermicompost is rich in all essential plant nutrients.
2. Provides excellent effect on overall plant growth, encourages the growth of new
Shoots / leaves and improves the quality and shelf life of the produce.
3. Vermicompost is free flowing, easy to apply, handle and store and does not have bad odor.
4. It improves soil structure, texture, aeration, and water holding capacity and prevents soil
erosion.
5. Vermicompost is rich in beneficial micro flora such as a fixers, P- solubilizers,
6. Cellulose decomposing micro-flora etc in addition to improve soil environment.
7. Vermicompost contains earthworm cocoons and increases the population and
Activity of earthworm in the soil.
8. It contains valuable vitamins, enzymes and hormones like auxins, gibberellins etc
9. It prevents nutrient losses and increases the use efficiency of chemical fertilizers.
10. Vermicompost is free from pathogens, toxic elements, weed seeds etc.
11. Vermicompost minimizes the incidence of pest and diseases.
12. It enhances the decomposition of organic matter in soil.
Tool of Vermicomposting (Earthworm):
Earthworms belong to the phylum Annelida; class Oligochaeta, which consists of over 7000 species.
Their bodies are long & tube like, tapering on both ends and commonly ranging length from one to six
inches. Certain Australian earthworms are several feet long. Earthworms are function as either a male
or a female during reproduction. Self-fertilization does not occur.
Internal anatomy of an earthworm (lateral section): Internal anatomy of an earthworm (ateral
section): small, long, cylindrical animal without legs or hard body parts.
Mouth cavity: Entrance to the digestive tract of an earthworm.
Brain: Brain of an earthworm.
Pharynx: Part of the digestive tract of an earthworm just after its mouth.
Esophagus: Part of the digestive tract of an earthworm between the pharynx and the crop.
Dorsal blood vessel: Blood vessel situated in the front part of an earthworm.
Seminal receptacle: Pocket related to the semen of an earthworm.
Seminal vesicles: Small hollow organs that carry the semen of an earthworm.
50 J. Chem. Bio. Phy. Sci. Sec. D, November 2016 – January 2017; Vol.7, No.1; 049-065.
Vermiculture has been the primary focus at Maharashtra Agricultural Bioteks in India, an
organization which has initiated both commercial and educational ventures to promote Vermiculture.
In 1985, Maharashtra Agricultural Bioteks was formed and established a small plant to manufacture
vermicompost from agricultural waste. In 1991-92, Maharashtra Bioteks and the India Department of
Science and Technology promoted the adoption of vermicompost technology in 13 states in India. The
group has also established a vermicompost unit with Chitrakoot Gramodaya University, Madhya
Pradesh which produces five tons of vermicompost per month.
Advantages of vermicompost:
1. Vermicompost is rich in all essential plant nutrients.
2. Provides excellent effect on overall plant growth, encourages the growth of new
Shoots / leaves and improves the quality and shelf life of the produce.
3. Vermicompost is free flowing, easy to apply, handle and store and does not have bad odor.
4. It improves soil structure, texture, aeration, and water holding capacity and prevents soil
erosion.
5. Vermicompost is rich in beneficial micro flora such as a fixers, P- solubilizers,
6. Cellulose decomposing micro-flora etc in addition to improve soil environment.
7. Vermicompost contains earthworm cocoons and increases the population and
Activity of earthworm in the soil.
8. It contains valuable vitamins, enzymes and hormones like auxins, gibberellins etc
9. It prevents nutrient losses and increases the use efficiency of chemical fertilizers.
10. Vermicompost is free from pathogens, toxic elements, weed seeds etc.
11. Vermicompost minimizes the incidence of pest and diseases.
12. It enhances the decomposition of organic matter in soil.
Tool of Vermicomposting (Earthworm):
Earthworms belong to the phylum Annelida; class Oligochaeta, which consists of over 7000 species.
Their bodies are long & tube like, tapering on both ends and commonly ranging length from one to six
inches. Certain Australian earthworms are several feet long. Earthworms are function as either a male
or a female during reproduction. Self-fertilization does not occur.
Internal anatomy of an earthworm (lateral section): Internal anatomy of an earthworm (ateral
section): small, long, cylindrical animal without legs or hard body parts.
Mouth cavity: Entrance to the digestive tract of an earthworm.
Brain: Brain of an earthworm.
Pharynx: Part of the digestive tract of an earthworm just after its mouth.
Esophagus: Part of the digestive tract of an earthworm between the pharynx and the crop.
Dorsal blood vessel: Blood vessel situated in the front part of an earthworm.
Seminal receptacle: Pocket related to the semen of an earthworm.
Seminal vesicles: Small hollow organs that carry the semen of an earthworm.
An Outlook … Kalam and Ahmad.
51 J. Chem. Bio. Phy. Sci. Sec. D, November 2016 – January 2017; Vol.7, No.1; 049-065.
Crop: Bulge of the esophagus of an earthworm.
Gizzard: Pocket used as the stomach of an earthworm.
Nephridia: Organ of an earthworm that performs the functions of kidneys.
Lateral heart: Blood-pumping organ of an earthworm.
Digestive System: The earthworm has a simple digestive system compared to other creatures. It
possesses an esophagus, which serves the same purpose as it does in higher animals: to carry food.
However, the worm esophagus leads to the crop and gizzard rather than to a stomach (which
earthworms do not possess). The crop and gizzard combined serve the purpose of a human stomach.
Gizzard’s main function is to grind and digest tough food through some digestive enzymes. The crop
holds food while the gizzard grinds it down mechanically to increase the surface area for the chemical
processing. Actually mineral matter accumulates in the gizzard, and ultimately the gizzard grinds the
particles. The gizzard leads into the intestine, which digests and absorbs the nutrients and it follows
the length of the earthworm’s body to the anus.
This passage from mouth -> esophagus -> crop -> gizzard -> anus constitutes the entire digestive
system.
Vermicast: Vermicast is produced by the feeding action of earthworms. Earthworms ingest organic
matter, fragmenting and grinding it into a finely divided peat like material with high porosity,
aeration, drainage and water holding capacity. Mucus type substance coated on each particle that
increases aeration in the soil, provides excellent water retention properties and improves the drainage
in heavy soils. This process enhances microbial activity and accelerates the rate of decomposition.
This leads to a humification effect where unstable organic matter or decomposing plant and animal
matter is oxidized and stabilized. Microbially, vermicast contains a far more diverse microbial
population (such as Azotobacter, PGPR, PSB, Actinomycetes) than other composts.
Microorganisms play an important part in soil fertility, they not only mineralize complex substances
into plant available nutrients but bacteria in the earth worms' digestive system also synthesize a whole
series of biologically active substances including plant growth regulators. Earth worms promote the
production of plant hormones, auxins, gibberellins and cytokinins from organic waste dramatically.
These hormones are dose significant and play a fundamental role in plant metabolism as well as plant
growth, development and crop quality significantly. Vermicast is humus rich. The breakdown of
organic material by earth worms accelerates the humification of organic matter.
The humic and fulvic acids produced in this process have been proven to stimulate plant growth.
Humic acids are large complex molecules. Partial oxidation of humic acids allows bonding sites for
plant nutrients including calcium and magnesium. Humic acids are produced by the breakdown of
organic matter by microorganisms. They are generally negatively charged so attracting positive ions,
for example calcium. Humic substances promote the conversion of a number of elements into forms
available to plants, of particular importance is phosphate. Phosphate reacts with other minerals in the
soil, (particularly iron and aluminium) and becomes locked or unavailable to plants. Humic acids help
substitute iron and aluminium with other elements e.g. calcium making phosphate plant available.
As a fertiliser, vermicast contains nutrients in a form that are readily taken up by plants, such as
nitrates, exchangeable phosphorous, soluble potassium, calcium and magnesium. A typical
breakdown of vermicast is as follows:
pH N P K S Mg Ca CEC (Me/100g) Organic Matter (humus)
6.6 2.3 3.0 0.6 0.1 0.65 8.6 52.75 20%
51 J. Chem. Bio. Phy. Sci. Sec. D, November 2016 – January 2017; Vol.7, No.1; 049-065.
Crop: Bulge of the esophagus of an earthworm.
Gizzard: Pocket used as the stomach of an earthworm.
Nephridia: Organ of an earthworm that performs the functions of kidneys.
Lateral heart: Blood-pumping organ of an earthworm.
Digestive System: The earthworm has a simple digestive system compared to other creatures. It
possesses an esophagus, which serves the same purpose as it does in higher animals: to carry food.
However, the worm esophagus leads to the crop and gizzard rather than to a stomach (which
earthworms do not possess). The crop and gizzard combined serve the purpose of a human stomach.
Gizzard’s main function is to grind and digest tough food through some digestive enzymes. The crop
holds food while the gizzard grinds it down mechanically to increase the surface area for the chemical
processing. Actually mineral matter accumulates in the gizzard, and ultimately the gizzard grinds the
particles. The gizzard leads into the intestine, which digests and absorbs the nutrients and it follows
the length of the earthworm’s body to the anus.
This passage from mouth -> esophagus -> crop -> gizzard -> anus constitutes the entire digestive
system.
Vermicast: Vermicast is produced by the feeding action of earthworms. Earthworms ingest organic
matter, fragmenting and grinding it into a finely divided peat like material with high porosity,
aeration, drainage and water holding capacity. Mucus type substance coated on each particle that
increases aeration in the soil, provides excellent water retention properties and improves the drainage
in heavy soils. This process enhances microbial activity and accelerates the rate of decomposition.
This leads to a humification effect where unstable organic matter or decomposing plant and animal
matter is oxidized and stabilized. Microbially, vermicast contains a far more diverse microbial
population (such as Azotobacter, PGPR, PSB, Actinomycetes) than other composts.
Microorganisms play an important part in soil fertility, they not only mineralize complex substances
into plant available nutrients but bacteria in the earth worms' digestive system also synthesize a whole
series of biologically active substances including plant growth regulators. Earth worms promote the
production of plant hormones, auxins, gibberellins and cytokinins from organic waste dramatically.
These hormones are dose significant and play a fundamental role in plant metabolism as well as plant
growth, development and crop quality significantly. Vermicast is humus rich. The breakdown of
organic material by earth worms accelerates the humification of organic matter.
The humic and fulvic acids produced in this process have been proven to stimulate plant growth.
Humic acids are large complex molecules. Partial oxidation of humic acids allows bonding sites for
plant nutrients including calcium and magnesium. Humic acids are produced by the breakdown of
organic matter by microorganisms. They are generally negatively charged so attracting positive ions,
for example calcium. Humic substances promote the conversion of a number of elements into forms
available to plants, of particular importance is phosphate. Phosphate reacts with other minerals in the
soil, (particularly iron and aluminium) and becomes locked or unavailable to plants. Humic acids help
substitute iron and aluminium with other elements e.g. calcium making phosphate plant available.
As a fertiliser, vermicast contains nutrients in a form that are readily taken up by plants, such as
nitrates, exchangeable phosphorous, soluble potassium, calcium and magnesium. A typical
breakdown of vermicast is as follows:
pH N P K S Mg Ca CEC (Me/100g) Organic Matter (humus)
6.6 2.3 3.0 0.6 0.1 0.65 8.6 52.75 20%
An Outlook … Kalam and Ahmad.
52 J. Chem. Bio. Phy. Sci. Sec. D, November 2016 – January 2017; Vol.7, No.1; 049-065.
Vermicast vs. Chemical Fertilizers in Soil
Criteria for
Comparison
Chemical Fertilizers Vermicast
Macro nutrient
contents
Mostly contains only one (N
in urea) or at the most two (N
& P in DAP) nutrients
Contains all i.e. nitrogen (N), phosphorus (P)
& potassium (K) in sufficient quantities
Secondary nutrient
contents
Not available Calcium, magnesium & sulphur is available in
required quantities
Micro nutrient
contents
Not available Zinc, boron, manganese, iron , copper,
molybdenum and chlorine also present
pH balancing Disturb soil pH to create
salinity and alkalinity
conditions
Helps in the control of soil pH and checks the
salinity and alkalinity in soil
Organic carbon Not available Very high organic carbon and humus contents
improves soil characteristics
Moisture retention
capacity
Reduces moisture retention
capacity,
Increases moistures retention capacity of the
soil
Soil Texture Damages soil texture to reduce
aeration
Improves soil texture for better aeration
Beneficial bacteria
& fungi
Reduces biological activities
and thus the fertility is
impaired
Very high biological life improves the soil
fertility and productivity on sustainable basis
Vermicompost Technology (Preparation in Flow Chart)
a. Keep the 100 kg. Raw materials (organic matter) in raised cemented chamber.
b. Take 16 kilograms of cattle dung and mix it with 25 liters of water.
c. Mix this mixture with 100 Kilograms of raw material.
d. Keep this mixture moist and stir once every week for five weeks. It will be partially decomposed.
e. Make four raised chambers measuring 3ft. x 3ft. x 3 ft. with cemented floor.
f. Fix an asbestos sheet six foot above for protection from rains.
52 J. Chem. Bio. Phy. Sci. Sec. D, November 2016 – January 2017; Vol.7, No.1; 049-065.
Vermicast vs. Chemical Fertilizers in Soil
Criteria for
Comparison
Chemical Fertilizers Vermicast
Macro nutrient
contents
Mostly contains only one (N
in urea) or at the most two (N
& P in DAP) nutrients
Contains all i.e. nitrogen (N), phosphorus (P)
& potassium (K) in sufficient quantities
Secondary nutrient
contents
Not available Calcium, magnesium & sulphur is available in
required quantities
Micro nutrient
contents
Not available Zinc, boron, manganese, iron , copper,
molybdenum and chlorine also present
pH balancing Disturb soil pH to create
salinity and alkalinity
conditions
Helps in the control of soil pH and checks the
salinity and alkalinity in soil
Organic carbon Not available Very high organic carbon and humus contents
improves soil characteristics
Moisture retention
capacity
Reduces moisture retention
capacity,
Increases moistures retention capacity of the
soil
Soil Texture Damages soil texture to reduce
aeration
Improves soil texture for better aeration
Beneficial bacteria
& fungi
Reduces biological activities
and thus the fertility is
impaired
Very high biological life improves the soil
fertility and productivity on sustainable basis
Vermicompost Technology (Preparation in Flow Chart)
a. Keep the 100 kg. Raw materials (organic matter) in raised cemented chamber.
b. Take 16 kilograms of cattle dung and mix it with 25 liters of water.
c. Mix this mixture with 100 Kilograms of raw material.
d. Keep this mixture moist and stir once every week for five weeks. It will be partially decomposed.
e. Make four raised chambers measuring 3ft. x 3ft. x 3 ft. with cemented floor.
f. Fix an asbestos sheet six foot above for protection from rains.
An Outlook … Kalam and Ahmad.
53 J. Chem. Bio. Phy. Sci. Sec. D, November 2016 – January 2017; Vol.7, No.1; 049-065.
g. Cover it with a perforated cover (Metallic) having two mm holes.
h. Keep the partially decomposed matter in these chambers at least 1inch below the top.
i. Put the mixture of 4000 there earthworm (Eudrilus eugeniae: Eisenia feotida: Perionyx excavatus=
1:1:1) over the surface of partially decomposed organic matter.
j. Spray little water once in three days.
k. These worms will eat the partially decomposed organic matter and put out the excreta as
vermicompost.
l. After every ten days by hand remove the vermicompost (excreta).
m. The vermicompost is sieved by use of one mm sieve and spread in thin layer for air drying.
n. Matter which could not be sieved is placed back on partially decomposed organic matter for
breakdown by earthworms.
o. In about one month all the partially decomposed organic matter gets converted into vermicompost.
p. Well air dried vermicompost is Packed in 1 to 5 Kg in polythene bags, and airtight.
DESCRIPTION OF VAT
Different types of vat are used.
53 J. Chem. Bio. Phy. Sci. Sec. D, November 2016 – January 2017; Vol.7, No.1; 049-065.
g. Cover it with a perforated cover (Metallic) having two mm holes.
h. Keep the partially decomposed matter in these chambers at least 1inch below the top.
i. Put the mixture of 4000 there earthworm (Eudrilus eugeniae: Eisenia feotida: Perionyx excavatus=
1:1:1) over the surface of partially decomposed organic matter.
j. Spray little water once in three days.
k. These worms will eat the partially decomposed organic matter and put out the excreta as
vermicompost.
l. After every ten days by hand remove the vermicompost (excreta).
m. The vermicompost is sieved by use of one mm sieve and spread in thin layer for air drying.
n. Matter which could not be sieved is placed back on partially decomposed organic matter for
breakdown by earthworms.
o. In about one month all the partially decomposed organic matter gets converted into vermicompost.
p. Well air dried vermicompost is Packed in 1 to 5 Kg in polythene bags, and airtight.
DESCRIPTION OF VAT
Different types of vat are used.
An Outlook … Kalam and Ahmad.
54 J. Chem. Bio. Phy. Sci. Sec. D, November 2016 – January 2017; Vol.7, No.1; 049-065.
Container method: Suitable for household purpose. Container of 1 m x 1 m x 0.75 m should contain
about 1000 worms. Container can be used provided it has good drainage.
Heap (Bed) Method:
Apart from this there are other types of bin that are used as commercial farm units. These are (a) Can-
O-Worms™ (b) Worm-A-Way® (c) Worm-A-Roo™.
(a) Can-O-Worms™
(b) Worm-A-Way®
(c) Worm-A-Roo™
Substrate (Raw Materials): Materials that is required for vermicomposting-
(A) 100 Kilograms of organic matter such as-
Suitable for both commercial and small farm units. Size of the shed varies
depending upon the availability of raw material and production requirement.
Small unit - 8m x 4m x 4m (10 tons production)
Big unit - 30m x 8m x 4m (2sheeds 100 tones production)
- Most popular
- Enclosed tier system
- Bottom catch tray & spigot
- Stackable mesh trays
- Worms migrate vertically
- asy to harvest castings
- $130.00 incl. shipping
-Plastic
-Ventilated
-Several sizes
-Lightweight
-$90-$100 incl. Worms & shipping
-Double bin system
-Plastic
-“Migration device”
-Lightweight
-$140-$170 incl. Supplies, worms, and shipping
54 J. Chem. Bio. Phy. Sci. Sec. D, November 2016 – January 2017; Vol.7, No.1; 049-065.
Container method: Suitable for household purpose. Container of 1 m x 1 m x 0.75 m should contain
about 1000 worms. Container can be used provided it has good drainage.
Heap (Bed) Method:
Apart from this there are other types of bin that are used as commercial farm units. These are (a) Can-
O-Worms™ (b) Worm-A-Way® (c) Worm-A-Roo™.
(a) Can-O-Worms™
(b) Worm-A-Way®
(c) Worm-A-Roo™
Substrate (Raw Materials): Materials that is required for vermicomposting-
(A) 100 Kilograms of organic matter such as-
Suitable for both commercial and small farm units. Size of the shed varies
depending upon the availability of raw material and production requirement.
Small unit - 8m x 4m x 4m (10 tons production)
Big unit - 30m x 8m x 4m (2sheeds 100 tones production)
- Most popular
- Enclosed tier system
- Bottom catch tray & spigot
- Stackable mesh trays
- Worms migrate vertically
- asy to harvest castings
- $130.00 incl. shipping
-Plastic
-Ventilated
-Several sizes
-Lightweight
-$90-$100 incl. Worms & shipping
-Double bin system
-Plastic
-“Migration device”
-Lightweight
-$140-$170 incl. Supplies, worms, and shipping
An Outlook … Kalam and Ahmad.
55 J. Chem. Bio. Phy. Sci. Sec. D, November 2016 – January 2017; Vol.7, No.1; 049-065.
1. Crop residues 5. Hotel refuse
2. Weed biomass 6. Waste from agro-industries
3. Vegetable waste 7. Biodegradable portion of urban and rural wastes.
4. Leaf litter 8. House organic waste
(B) Cattle dung about 16 Kilograms
(C) Earthworms species:
(a.) Eudrilus eugeniae
(b). Eisenia feotida
(c). Perionyx excavatus. About 4000.
PROCEDURE
Partial Decomposing of Raw Material:
a. Keep the 100 kg. Raw materials (organic matter) in raised cemented chamber.
b. Take 16 kilograms of cattle dung and mix it with 25 liters of water.
c. Mix this mixture with 100 Kilograms of raw material.
d. Keep this mixture moist and stir once every week for five weeks. It will be partially
decomposed.
Making of vermicompost from partially decomposed raw material:
a. Make four raised chambers measuring 3ft. x 3ft. x 3 ft. with cemented floor.
b. Fix an asbestos sheet six foot above for protection from rains.
c. Keep the partially decomposed matter in these chambers at least one inch below the top.
d. Cover it with a perforated cover (Metallic) having two mm holes.
e. Put the mixture of 4000 there earthworm (1:1:1) over the surface of partially decomposed
organic matter.
f. Keep it moist by little water spray. These worms will eat the partially decomposed organic
matter and put out the excreta as vermicompost. Spray little water once in three days.
Processing of the vermicompost:
a. After every ten days by hand remove the vermicompost (excreta).
b. Keep this vermicompost in heap in order to separate the very small earthworms. In 24 hours
these earthworms comes to the bottom part of vermicompost, which are then removed and
placed on the partially decomposed organic matter.
c. The vermicompost is sieved by use of one mm sieve and spread in thin layer for air drying.
d. Matter which could not be sieved is placed back on partially decomposed organic matter for
breakdown by earthworms.
e. In about one month all the partially decomposed organic matter gets converted into
vermicompost.
55 J. Chem. Bio. Phy. Sci. Sec. D, November 2016 – January 2017; Vol.7, No.1; 049-065.
1. Crop residues 5. Hotel refuse
2. Weed biomass 6. Waste from agro-industries
3. Vegetable waste 7. Biodegradable portion of urban and rural wastes.
4. Leaf litter 8. House organic waste
(B) Cattle dung about 16 Kilograms
(C) Earthworms species:
(a.) Eudrilus eugeniae
(b). Eisenia feotida
(c). Perionyx excavatus. About 4000.
PROCEDURE
Partial Decomposing of Raw Material:
a. Keep the 100 kg. Raw materials (organic matter) in raised cemented chamber.
b. Take 16 kilograms of cattle dung and mix it with 25 liters of water.
c. Mix this mixture with 100 Kilograms of raw material.
d. Keep this mixture moist and stir once every week for five weeks. It will be partially
decomposed.
Making of vermicompost from partially decomposed raw material:
a. Make four raised chambers measuring 3ft. x 3ft. x 3 ft. with cemented floor.
b. Fix an asbestos sheet six foot above for protection from rains.
c. Keep the partially decomposed matter in these chambers at least one inch below the top.
d. Cover it with a perforated cover (Metallic) having two mm holes.
e. Put the mixture of 4000 there earthworm (1:1:1) over the surface of partially decomposed
organic matter.
f. Keep it moist by little water spray. These worms will eat the partially decomposed organic
matter and put out the excreta as vermicompost. Spray little water once in three days.
Processing of the vermicompost:
a. After every ten days by hand remove the vermicompost (excreta).
b. Keep this vermicompost in heap in order to separate the very small earthworms. In 24 hours
these earthworms comes to the bottom part of vermicompost, which are then removed and
placed on the partially decomposed organic matter.
c. The vermicompost is sieved by use of one mm sieve and spread in thin layer for air drying.
d. Matter which could not be sieved is placed back on partially decomposed organic matter for
breakdown by earthworms.
e. In about one month all the partially decomposed organic matter gets converted into
vermicompost.
An Outlook … Kalam and Ahmad.
56 J. Chem. Bio. Phy. Sci. Sec. D, November 2016 – January 2017; Vol.7, No.1; 049-065.
f. After 10 days of drying, vermicompost is collected and put in a heap. The idea is to remove
any remaining earthworm. For this purpose about half Kilogram of cattle dung is put inside
this heap. In about two days all remaining earthworms collect in the cattle dung. This cattle
dung is removed and placed on the recharged matter.
Vermicompost is ready in 2-2.5 months. When it is ready, it’s back, lightweight and has no bed smell.
PACKING
Before packing, the vermicompost is dried in air well but not in sunlight. Because in sunlight moisture
content of vermicompost become very low and as a result market price will be very low.
It is packed in 25-50 kgs polythene bags and airtight, just like that of other inorganic chemical
fertilizers packed in polythene bags. Packing cost per packet is generally 15-20 Rs.
Necessary Precautions
a. For protection of vermicompost preparation chamber from rains, roofing is essential.
b. Covering of vermicompost chamber by perforated cover is essential for proper aeration and
protection from rodents, birds etc.
c. Maintain the moisture at 50-60 % level in the pit.
d. Temperature between 25-28 ºC.
e. Base material (FYM) should be partially decomposed.
f. Proper aeration should be provided without disturbing the worms.
a. G.Vermicompost is dried in air well before packing.
APPLICATION
Vermicompost can be used for all crops (agricultural, horticultural, ornamental and vegetable) at any
stage of crop development.
For agricultural crops: Vermicompost can be applied in agricultural crops by broadcasting when the
seedlings are 12 to 15 cms in height and irrigate the field.
Flower, Vegetables and fruit trees: Apply vermicompost around the base of the plant, at any stage
of development and cover with soil. Water regularly.
Generally agricultural use: 3-4 tonnes/ha
Fruit trees: 5-10 kg/tree
Vegetables: 3-4tonnes/ha
Flowers: 500-750 kg/ha
56 J. Chem. Bio. Phy. Sci. Sec. D, November 2016 – January 2017; Vol.7, No.1; 049-065.
f. After 10 days of drying, vermicompost is collected and put in a heap. The idea is to remove
any remaining earthworm. For this purpose about half Kilogram of cattle dung is put inside
this heap. In about two days all remaining earthworms collect in the cattle dung. This cattle
dung is removed and placed on the recharged matter.
Vermicompost is ready in 2-2.5 months. When it is ready, it’s back, lightweight and has no bed smell.
PACKING
Before packing, the vermicompost is dried in air well but not in sunlight. Because in sunlight moisture
content of vermicompost become very low and as a result market price will be very low.
It is packed in 25-50 kgs polythene bags and airtight, just like that of other inorganic chemical
fertilizers packed in polythene bags. Packing cost per packet is generally 15-20 Rs.
Necessary Precautions
a. For protection of vermicompost preparation chamber from rains, roofing is essential.
b. Covering of vermicompost chamber by perforated cover is essential for proper aeration and
protection from rodents, birds etc.
c. Maintain the moisture at 50-60 % level in the pit.
d. Temperature between 25-28 ºC.
e. Base material (FYM) should be partially decomposed.
f. Proper aeration should be provided without disturbing the worms.
a. G.Vermicompost is dried in air well before packing.
APPLICATION
Vermicompost can be used for all crops (agricultural, horticultural, ornamental and vegetable) at any
stage of crop development.
For agricultural crops: Vermicompost can be applied in agricultural crops by broadcasting when the
seedlings are 12 to 15 cms in height and irrigate the field.
Flower, Vegetables and fruit trees: Apply vermicompost around the base of the plant, at any stage
of development and cover with soil. Water regularly.
Generally agricultural use: 3-4 tonnes/ha
Fruit trees: 5-10 kg/tree
Vegetables: 3-4tonnes/ha
Flowers: 500-750 kg/ha
An Outlook … Kalam and Ahmad.
57 J. Chem. Bio. Phy. Sci. Sec. D, November 2016 – January 2017; Vol.7, No.1; 049-065.
Nutrient status of vermicompost
Organic carbon % 20.43 – 30.31
Total nitrogen % 1.80 – 2.05
Phosphorus % 1.32 – 1.93
Potassium % 1.28 – 1.50
Calcium % 3.0 – 4.5
Copper % 0.0027 – 0.0123
Iron % 0.3 – 0.7
Zinc % 0.028 – 0.036
Sulphur % Traces to 0.40
Magnesium % 0.4 – 0.7
Sodium % 0.02 – 0.30
Boron % 0.0034 – 0.0075
Manganese % 0.40
Aluminium % Traces to 0.071
Cobalt, Molybdenum Present in available form
Carbon : Nitrogen 14-15 : 1
pH 6.5-7.5
Vermiwash: Vermin wash is a liquid plant growth regulator, which contains high amount of
enzymes, Vitamins and hormones like auxins, gibberellins etc along with macro and micronutrients
Used as foliar spray.
Apparatus: An apparatus for making a vermiwash, the apparatus comprising:
1. A bucket for holding the particulate matter.
2. One stop cork.
3. Two hanging pot
The stop cork added to the lower most part of the bucket .One hanging pot hangs over the bucket
which contain a hole in such a way so that waterfalls drop by drop in bucket and another hanging pot
hangs lower part of the stop cork so that vermiwash produced in the bucket collected in the pot. Upper
hanging pot contain water one fiftieth of the size of the main container.
Water is poured into this container and allowed to gradually sprinkle on the bucket overnight. This
water percolates through the compost, the burrows of the earthworms and gets collected at the base.
The stop cork is opened the next day morning and the vermiwash is collected. The tap is then closed
and the suspended pot is refilled with water that evening to be collected again the following morning.
Dung pats and hay may be replaced periodically based on need. The entire set up may be emptied and
reset between 10 and 12 months of use.
Substrate (Raw Materials)
1. Broken bricks 4. Partially decomposed cow dung.
2. Pieces of stones. 5. Soil.
3. Sand. 6. 100-200 nos. of earthworms
7. A layer of paddy straw.
57 J. Chem. Bio. Phy. Sci. Sec. D, November 2016 – January 2017; Vol.7, No.1; 049-065.
Nutrient status of vermicompost
Organic carbon % 20.43 – 30.31
Total nitrogen % 1.80 – 2.05
Phosphorus % 1.32 – 1.93
Potassium % 1.28 – 1.50
Calcium % 3.0 – 4.5
Copper % 0.0027 – 0.0123
Iron % 0.3 – 0.7
Zinc % 0.028 – 0.036
Sulphur % Traces to 0.40
Magnesium % 0.4 – 0.7
Sodium % 0.02 – 0.30
Boron % 0.0034 – 0.0075
Manganese % 0.40
Aluminium % Traces to 0.071
Cobalt, Molybdenum Present in available form
Carbon : Nitrogen 14-15 : 1
pH 6.5-7.5
Vermiwash: Vermin wash is a liquid plant growth regulator, which contains high amount of
enzymes, Vitamins and hormones like auxins, gibberellins etc along with macro and micronutrients
Used as foliar spray.
Apparatus: An apparatus for making a vermiwash, the apparatus comprising:
1. A bucket for holding the particulate matter.
2. One stop cork.
3. Two hanging pot
The stop cork added to the lower most part of the bucket .One hanging pot hangs over the bucket
which contain a hole in such a way so that waterfalls drop by drop in bucket and another hanging pot
hangs lower part of the stop cork so that vermiwash produced in the bucket collected in the pot. Upper
hanging pot contain water one fiftieth of the size of the main container.
Water is poured into this container and allowed to gradually sprinkle on the bucket overnight. This
water percolates through the compost, the burrows of the earthworms and gets collected at the base.
The stop cork is opened the next day morning and the vermiwash is collected. The tap is then closed
and the suspended pot is refilled with water that evening to be collected again the following morning.
Dung pats and hay may be replaced periodically based on need. The entire set up may be emptied and
reset between 10 and 12 months of use.
Substrate (Raw Materials)
1. Broken bricks 4. Partially decomposed cow dung.
2. Pieces of stones. 5. Soil.
3. Sand. 6. 100-200 nos. of earthworms
7. A layer of paddy straw.
An Outlook … Kalam and Ahmad.
58 J. Chem. Bio. Phy. Sci. Sec. D, November 2016 – January 2017; Vol.7, No.1; 049-065.
PREPARATION
1. Take one big bucket and one mug.
2. Set up one stop cork on the lower most part of the bucket.
3. Put a layer of broken bricks, pieces of stones having thickness of 10-15 cm in the bucket.
4. Over this layer put another layer of sand having thickness of 10-15 cm.
5. Then put a layer of partially decomposed cow dung having 30-45 cm thickness over it.
6. Then put another layer of soil having 2-3 thicknesses.
7. Now open the stopcock of the bucket and when the materials taken in the bucket.
8. Then put 100-200 nos. of earthworms in the bucket.
9. After that, a layer of paddy straw having 6 cm thickness is given.
10. Now open the stopcock of the bucket and spray water regularly for a period of 7-8 days.
11. After 10 days the liquid vermin wash will be produced in the bucket.
12. Hang one pot with a bottom hole over the bucket in such a way so that water falls drop by drop.
13. Every day 4-5 liters of water is to be poured in the hanging pot.
14. Keep another pot under stop cork to collect the vermin wash. Every day 3-4 liters Vermin wash
can be collected.
18. Vermiwash Analysis Report.
Organic Carbon % 0.008 ± 0.001
Total Kjeldhal Nitrogen % 0.01±0.005
Available Phosphate % 1.69 ± 0.05
Potassium (ppm) 25 ± 2
Sodium (ppm) 8 ± 1
Calcium (ppm) 3 ± 1
Copper (ppm) 0.01 ± 0.001
Ferrous (ppm) 0.06 ± 0.001
Magnesium (ppm) 158.44 ± 23.42
Manganese (ppm) 0.58 ± 0.040
Zinc (ppm) 0.02 ± 0.001
Total Heterotrophs (CFU/ml) 1.79 x 103
Electro conductivity dS/m 0.25 ± 0.03
pH 7.48 ± 0.03
Application
1. Mix 1 liters of vermin wash with 7-10 liters of water and spray the solution in the leaf (upper and
lower side) in the evening at the growing the crop.
2. Mix 1 liter of vermin wash with 1 liter of cow urine and then add 10 liters of water to the vermin
urine solution and mixed thoroughly and keep it over night before spraying 50-60 liter of such
solution and to be sprayed in one bigha of land to control various crop diseases.
58 J. Chem. Bio. Phy. Sci. Sec. D, November 2016 – January 2017; Vol.7, No.1; 049-065.
PREPARATION
1. Take one big bucket and one mug.
2. Set up one stop cork on the lower most part of the bucket.
3. Put a layer of broken bricks, pieces of stones having thickness of 10-15 cm in the bucket.
4. Over this layer put another layer of sand having thickness of 10-15 cm.
5. Then put a layer of partially decomposed cow dung having 30-45 cm thickness over it.
6. Then put another layer of soil having 2-3 thicknesses.
7. Now open the stopcock of the bucket and when the materials taken in the bucket.
8. Then put 100-200 nos. of earthworms in the bucket.
9. After that, a layer of paddy straw having 6 cm thickness is given.
10. Now open the stopcock of the bucket and spray water regularly for a period of 7-8 days.
11. After 10 days the liquid vermin wash will be produced in the bucket.
12. Hang one pot with a bottom hole over the bucket in such a way so that water falls drop by drop.
13. Every day 4-5 liters of water is to be poured in the hanging pot.
14. Keep another pot under stop cork to collect the vermin wash. Every day 3-4 liters Vermin wash
can be collected.
18. Vermiwash Analysis Report.
Organic Carbon % 0.008 ± 0.001
Total Kjeldhal Nitrogen % 0.01±0.005
Available Phosphate % 1.69 ± 0.05
Potassium (ppm) 25 ± 2
Sodium (ppm) 8 ± 1
Calcium (ppm) 3 ± 1
Copper (ppm) 0.01 ± 0.001
Ferrous (ppm) 0.06 ± 0.001
Magnesium (ppm) 158.44 ± 23.42
Manganese (ppm) 0.58 ± 0.040
Zinc (ppm) 0.02 ± 0.001
Total Heterotrophs (CFU/ml) 1.79 x 103
Electro conductivity dS/m 0.25 ± 0.03
pH 7.48 ± 0.03
Application
1. Mix 1 liters of vermin wash with 7-10 liters of water and spray the solution in the leaf (upper and
lower side) in the evening at the growing the crop.
2. Mix 1 liter of vermin wash with 1 liter of cow urine and then add 10 liters of water to the vermin
urine solution and mixed thoroughly and keep it over night before spraying 50-60 liter of such
solution and to be sprayed in one bigha of land to control various crop diseases.
An Outlook … Kalam and Ahmad.
59 J. Chem. Bio. Phy. Sci. Sec. D, November 2016 – January 2017; Vol.7, No.1; 049-065.
Vermiculture: Vermiculture refers to the biomass production of earthworms in semi natural
conditions, providing suitable substrate and feed for them to remain active all through the year.
Culturing of earthworms started as entrepreneurship in the later half of the 20th C. to supply them to
fishing enthusiasts. Earthworms that can survive in captivity under semi natural conditions, tolerant to
wide ranges of substrates and to other physical parameters like pH, temperature, moisture and
physical disturbances can only be maintained as cultures. They should show good population growth
for culture propagation. These characters are found in very few species of earthworms and hence,
successful culturing is possible only with these earthworms irrespective of the place of their origin.
In USA and Canada culturing of earthworms started as entrepreneurship where as in India they were
maintained as cultures for different research activities. Since more than a decade, farmers, agro based
industries and urban households are culturing earthworms as the biological material for organic waste
management in India. Though, many research laboratories are involved in carrying out research on
this aspect.
Vermicomposting and Sustainable Environment: Vermicompost, like conventional compost,
provides many benefits to agricultural soil, including increased ability to retain moisture, better
nutrient-holding capacity, better soil structure, and higher levels of microbial activity. Apart from this
Vermicomposts have great potential in horticulture and agriculture crop production due to production
of plant growth regulators by the greatly increased microbial populations. These accelerate the
germination, growth, flowering and yields of plants independent of nutrient supply. Vermicomposts
also have potential, as solids or aqueous vermicompost extracts, in integrated pest management
programs, since one application suppresses soil-borne plant pathogens, plant parasitic nematodes as
well as numbers and reproduction of arthropod pests such as aphids, beetles and caterpillars.
Pesticide/Herbicide Detoxification by Earthworm: Organophosphate degrading enzymes have been
intensively investigated in microorganisms. Since they can be potentially utilized to detoxify
environmental pollutions such as industrial wastes and pesticides. Phosphotriesterase (PTE) is an
enzyme that is able to hydrolyze organophosphate triesters. PTE was first detected in the soil
microorganisms, Pseudomonas diminuta and Flavobacterium sp, which is capable of hydrolyzing
paraoxon and parathion at a high catalytic activity. In soil macroinvertibrate such as earthworms, PTE
system was found, to effectively hydrolyze organophosphate pesticides applied to soil.
Earthworms play an important role in the disposition of soil xenobiotics. Glutathione and glutathione-
s-transferase (GST) play a major role in cellular defense mechanisms. Glutathione and glutathione-s-
transferase (GST) are present in earthworm. Glutathione S-transferases have been intensively studied
for their involvement in herbicide detoxification. Several widely used herbicides, among them
alachlor, metolachlor, propachlor or fenoxaprop are detoxified in biological systems by the formation
of glutathione acetanilide conjugates. This conjugation is mediated by GST. Carboxylesterases (CbEs;
EC 3.1.1.1) are hydrolases that cleave carboxyl esters to yield the corresponding alcohol and
carboxylic acid. These enzymes participate in the detoxification of pyrethroid (PYD), carbamate (CB)
and some organophosphorus (OP) insecticides. This Carboxyl esterases is found in earthworms some
tissues and organs seminal vesicles, seminal receptacles, pharynx, crop, gizzard, anterior intestine
wall muscle and Intestinal tissues.
Phosphotriesterase (PTE) system :PTE activity appeared to be primarily localized in intestinal
tissues. The highest level of PTE activity was found in epithelial tissue. The native molecular weight
of earthworm PTE was 260 kDa and the isoelectric point was approximately
4
. The optimal pH was
approximately
9
.
59 J. Chem. Bio. Phy. Sci. Sec. D, November 2016 – January 2017; Vol.7, No.1; 049-065.
Vermiculture: Vermiculture refers to the biomass production of earthworms in semi natural
conditions, providing suitable substrate and feed for them to remain active all through the year.
Culturing of earthworms started as entrepreneurship in the later half of the 20th C. to supply them to
fishing enthusiasts. Earthworms that can survive in captivity under semi natural conditions, tolerant to
wide ranges of substrates and to other physical parameters like pH, temperature, moisture and
physical disturbances can only be maintained as cultures. They should show good population growth
for culture propagation. These characters are found in very few species of earthworms and hence,
successful culturing is possible only with these earthworms irrespective of the place of their origin.
In USA and Canada culturing of earthworms started as entrepreneurship where as in India they were
maintained as cultures for different research activities. Since more than a decade, farmers, agro based
industries and urban households are culturing earthworms as the biological material for organic waste
management in India. Though, many research laboratories are involved in carrying out research on
this aspect.
Vermicomposting and Sustainable Environment: Vermicompost, like conventional compost,
provides many benefits to agricultural soil, including increased ability to retain moisture, better
nutrient-holding capacity, better soil structure, and higher levels of microbial activity. Apart from this
Vermicomposts have great potential in horticulture and agriculture crop production due to production
of plant growth regulators by the greatly increased microbial populations. These accelerate the
germination, growth, flowering and yields of plants independent of nutrient supply. Vermicomposts
also have potential, as solids or aqueous vermicompost extracts, in integrated pest management
programs, since one application suppresses soil-borne plant pathogens, plant parasitic nematodes as
well as numbers and reproduction of arthropod pests such as aphids, beetles and caterpillars.
Pesticide/Herbicide Detoxification by Earthworm: Organophosphate degrading enzymes have been
intensively investigated in microorganisms. Since they can be potentially utilized to detoxify
environmental pollutions such as industrial wastes and pesticides. Phosphotriesterase (PTE) is an
enzyme that is able to hydrolyze organophosphate triesters. PTE was first detected in the soil
microorganisms, Pseudomonas diminuta and Flavobacterium sp, which is capable of hydrolyzing
paraoxon and parathion at a high catalytic activity. In soil macroinvertibrate such as earthworms, PTE
system was found, to effectively hydrolyze organophosphate pesticides applied to soil.
Earthworms play an important role in the disposition of soil xenobiotics. Glutathione and glutathione-
s-transferase (GST) play a major role in cellular defense mechanisms. Glutathione and glutathione-s-
transferase (GST) are present in earthworm. Glutathione S-transferases have been intensively studied
for their involvement in herbicide detoxification. Several widely used herbicides, among them
alachlor, metolachlor, propachlor or fenoxaprop are detoxified in biological systems by the formation
of glutathione acetanilide conjugates. This conjugation is mediated by GST. Carboxylesterases (CbEs;
EC 3.1.1.1) are hydrolases that cleave carboxyl esters to yield the corresponding alcohol and
carboxylic acid. These enzymes participate in the detoxification of pyrethroid (PYD), carbamate (CB)
and some organophosphorus (OP) insecticides. This Carboxyl esterases is found in earthworms some
tissues and organs seminal vesicles, seminal receptacles, pharynx, crop, gizzard, anterior intestine
wall muscle and Intestinal tissues.
Phosphotriesterase (PTE) system :PTE activity appeared to be primarily localized in intestinal
tissues. The highest level of PTE activity was found in epithelial tissue. The native molecular weight
of earthworm PTE was 260 kDa and the isoelectric point was approximately
4
. The optimal pH was
approximately
9
.
An Outlook … Kalam and Ahmad.
60 J. Chem. Bio. Phy. Sci. Sec. D, November 2016 – January 2017; Vol.7, No.1; 049-065.
This enzyme was shown to bioactivate pesticides containing p-nitrophenyl moiety such as p-
nitrophenylphosphate, bis (p-nitrophenyl) phosphate. Parathion and paraoxon by releasing p-
nitrophenol as metabolite. In the earthworm, it has been reported that paraoxon hydrolase, a PTE was
able to hydrolyze paraoxon to produce diethylphosphate and p-nitrophenol in vivo and in vitro.
The presence of EGTA and EDTA completely abolished the activity and replacement of Ca2+ ion
restored activity to greater than 95%, suggesting that Ca2+ ion is essential to maintain the activity.
Figure 1: Histological distribution of phosphotriesterase activity in Eisenia Andrei.
Each tissue was homogenized in 50mM Tris-HCl buffer (pH 8.5) containing 0.1% Triton X-100 and
2mM CaCl2. After centrifugation at 13,000 g for 60 min, the resulting supernatant was used as enzyme
source of phosphotriesterase activity. Phosphotriesterase activity was measured by monitoring the
absorbance at 400 nm of p-nitrophenol produced when 1mM paraoxon was hydrolyzed into
diethylphosphate and p-nitrophenol. As shown in the Figure 1, all tissues tested exhibited PTE
activity. However over 90% of PTE activity appeared to be associated with gut tissues such as
chloragogue and epithelial tissue. The highest level PTE activity was found in the epithelial tissue.
Ch-Chloragogue tissue; Ep-epithelial tissue of gut; Ex-Extra-gut tissue.
Figure 2: Determination of isoelectric point of the PTE from Eisenia Andrei.
60 J. Chem. Bio. Phy. Sci. Sec. D, November 2016 – January 2017; Vol.7, No.1; 049-065.
This enzyme was shown to bioactivate pesticides containing p-nitrophenyl moiety such as p-
nitrophenylphosphate, bis (p-nitrophenyl) phosphate. Parathion and paraoxon by releasing p-
nitrophenol as metabolite. In the earthworm, it has been reported that paraoxon hydrolase, a PTE was
able to hydrolyze paraoxon to produce diethylphosphate and p-nitrophenol in vivo and in vitro.
The presence of EGTA and EDTA completely abolished the activity and replacement of Ca2+ ion
restored activity to greater than 95%, suggesting that Ca2+ ion is essential to maintain the activity.
Figure 1: Histological distribution of phosphotriesterase activity in Eisenia Andrei.
Each tissue was homogenized in 50mM Tris-HCl buffer (pH 8.5) containing 0.1% Triton X-100 and
2mM CaCl2. After centrifugation at 13,000 g for 60 min, the resulting supernatant was used as enzyme
source of phosphotriesterase activity. Phosphotriesterase activity was measured by monitoring the
absorbance at 400 nm of p-nitrophenol produced when 1mM paraoxon was hydrolyzed into
diethylphosphate and p-nitrophenol. As shown in the Figure 1, all tissues tested exhibited PTE
activity. However over 90% of PTE activity appeared to be associated with gut tissues such as
chloragogue and epithelial tissue. The highest level PTE activity was found in the epithelial tissue.
Ch-Chloragogue tissue; Ep-epithelial tissue of gut; Ex-Extra-gut tissue.
Figure 2: Determination of isoelectric point of the PTE from Eisenia Andrei.
An Outlook … Kalam and Ahmad.
61 J. Chem. Bio. Phy. Sci. Sec. D, November 2016 – January 2017; Vol.7, No.1; 049-065.
After focusing, the pH gradient (open circle) was normally established through a range from 2.5 to
10.0. The pH value was determined as the mean of three fractions, the distribution of
phosphotriesterase activity in each fraction was determined (closed circle).An aliquot of 200 micro
liter of each fraction was added to assay buffer (0.2 M, pH 8.5 Tris-HCl contain 2mM CaCl2). Note a
distinct peak at fraction no 3 for which the pH was 4. Therefore earthworm PTE appeared to be acidic
protein.
Figure 3: pH profile of phosphotriesterase activity from Eisenia andrei- At pH 9 the activity was
maximal. Therefore the enzyme could be alkaline phosphotriesterase.
Figure 4: Double reciprocal plot of phosphotriesterase activity of the earthworm
Eisenia andrei with paraoxon as substrate at pH 8.5.
Activity (v) is expressed as nmol p-nitrophenol produced/min/mg protein. Data points represent the
mean SD of triplicate determinations. The range of substrate concentration was 0.25 to 2 mM. The
double reciprocal plot yielded apparent Km and Vmax of 2.4mM and 4.8nM/min/mg respectively.
From the above experiment it is concluded that PTE system of earthworm has the ability to detoxify
pesticides.
Heavy Metal Tolerance by Earthworm: The earthworm Lumbricus rubellus takes up and retains
lead from soil containing a high concentration of lead. Irrespective of the copper, zinc and manganese
concentrations in the soil, these metals appear to be regulated in the tissues.
61 J. Chem. Bio. Phy. Sci. Sec. D, November 2016 – January 2017; Vol.7, No.1; 049-065.
After focusing, the pH gradient (open circle) was normally established through a range from 2.5 to
10.0. The pH value was determined as the mean of three fractions, the distribution of
phosphotriesterase activity in each fraction was determined (closed circle).An aliquot of 200 micro
liter of each fraction was added to assay buffer (0.2 M, pH 8.5 Tris-HCl contain 2mM CaCl2). Note a
distinct peak at fraction no 3 for which the pH was 4. Therefore earthworm PTE appeared to be acidic
protein.
Figure 3: pH profile of phosphotriesterase activity from Eisenia andrei- At pH 9 the activity was
maximal. Therefore the enzyme could be alkaline phosphotriesterase.
Figure 4: Double reciprocal plot of phosphotriesterase activity of the earthworm
Eisenia andrei with paraoxon as substrate at pH 8.5.
Activity (v) is expressed as nmol p-nitrophenol produced/min/mg protein. Data points represent the
mean SD of triplicate determinations. The range of substrate concentration was 0.25 to 2 mM. The
double reciprocal plot yielded apparent Km and Vmax of 2.4mM and 4.8nM/min/mg respectively.
From the above experiment it is concluded that PTE system of earthworm has the ability to detoxify
pesticides.
Heavy Metal Tolerance by Earthworm: The earthworm Lumbricus rubellus takes up and retains
lead from soil containing a high concentration of lead. Irrespective of the copper, zinc and manganese
concentrations in the soil, these metals appear to be regulated in the tissues.
An Outlook … Kalam and Ahmad.
62 J. Chem. Bio. Phy. Sci. Sec. D, November 2016 – January 2017; Vol.7, No.1; 049-065.
In soil containing high concentrations of zinc and calcium there are species differences in tissue metal
content. L. rubellus contained the highest amounts of zinc and manganese, Dendrobaena veneta the
highest concentration of cadmium and the highest mean lead content was found in Eiseniella
tetraedra. The concentration of soil calcium and copper, together with species differences in tolerance
to heavy metals, are considered as factors which may influence heavy metal accumulation.
Earthworms concentrate cadmium from the environment at higher levels than any other metal studied.
Bioaccumulation is usually based on a summation of the amount of metal adsorbed to the body wall
and absorbed into the body. The relative proportions of metal adsorption and absorption are usually
not quantified. The distinction between adsorbed and absorbed metals was investigated when exposed
to metals for 2 weeks. The earthworm Lumbricus rubellus was chosen as representative for organisms
mainly taking up metals via the dermal route. Cross-sections of whole animals were made using a
cryostat and accumulated metals were localized by means of an autoradiography. Radiolabels were
used to determine the distribution of metals over the different organs and to distinguish between
adsorption and absorption. Cd in the earthworm was mainly found in tissues of the chloragogenous
region, whereas Zn was also found in various other organs and in the connective tissue. Adsorbed
amounts of Cd and Zn were negligible compared to internalized Cd and Zn concentrations.
Future Perspective
1. Highlights the critical beneficial role of earthworms for plant and soil health and their important
role in organic waste management
2. Presents in-depth discussion of earthworm biogeography, diversity, taxonomy, and systematic.
3. Provides full coverage of feeding behavior, food preferences, dietary requirements, and the
interaction between earthworms and microbial communities.
4. Includes information on the impacts of climate, human activities, soil properties, predation,
disease and parasitism, and competition upon earthworm ecology.
5. It also controls soil as well as environmental pollution. So we have to know how it helps to
control soil as well as environmental pollution.
6. Earthworms effectively harness the beneficial soil micro flora, destroy soil pathogen and convert
organic wastes in to vitamins, enzymes, antibiotics, PGH, protein rich products and others organic
compounds. More information is needed in this field.
7. Offers new perspectives on vermicomposting research.
CONCLUSION
“Nobody and nothing can be compared with earthworms in their positive influence on the whole
living Nature. They create soil and everything that lives in it. They are the most numerous animals on
Earth and the main creatures converting all organic matter into soil humus providing soil’s fertility
and biosphere’s functions: disinfecting, neutralizing, protective and productive.”
(Anatoly M. Igonin21, Ph. D., Professor at the Vladimir Pedagogical University, Vladimir, Russia, as
quoted in Casting Call 9(2), Aug 2004.)
Aristotle called worms the “intestines of the earth” and Charles Darwin wrote a book on worms and
their activities, in which he stated that there may not be any other creature that has played so
important a role in the history of life on earth (Bogdanov
36
)(Pl. delete). There can be little doubt that
humankind’s relationship with worms is vital and needs to be nurtured and expanded. The following
sections touch on some of the most important areas in which our natural environment can be
62 J. Chem. Bio. Phy. Sci. Sec. D, November 2016 – January 2017; Vol.7, No.1; 049-065.
In soil containing high concentrations of zinc and calcium there are species differences in tissue metal
content. L. rubellus contained the highest amounts of zinc and manganese, Dendrobaena veneta the
highest concentration of cadmium and the highest mean lead content was found in Eiseniella
tetraedra. The concentration of soil calcium and copper, together with species differences in tolerance
to heavy metals, are considered as factors which may influence heavy metal accumulation.
Earthworms concentrate cadmium from the environment at higher levels than any other metal studied.
Bioaccumulation is usually based on a summation of the amount of metal adsorbed to the body wall
and absorbed into the body. The relative proportions of metal adsorption and absorption are usually
not quantified. The distinction between adsorbed and absorbed metals was investigated when exposed
to metals for 2 weeks. The earthworm Lumbricus rubellus was chosen as representative for organisms
mainly taking up metals via the dermal route. Cross-sections of whole animals were made using a
cryostat and accumulated metals were localized by means of an autoradiography. Radiolabels were
used to determine the distribution of metals over the different organs and to distinguish between
adsorption and absorption. Cd in the earthworm was mainly found in tissues of the chloragogenous
region, whereas Zn was also found in various other organs and in the connective tissue. Adsorbed
amounts of Cd and Zn were negligible compared to internalized Cd and Zn concentrations.
Future Perspective
1. Highlights the critical beneficial role of earthworms for plant and soil health and their important
role in organic waste management
2. Presents in-depth discussion of earthworm biogeography, diversity, taxonomy, and systematic.
3. Provides full coverage of feeding behavior, food preferences, dietary requirements, and the
interaction between earthworms and microbial communities.
4. Includes information on the impacts of climate, human activities, soil properties, predation,
disease and parasitism, and competition upon earthworm ecology.
5. It also controls soil as well as environmental pollution. So we have to know how it helps to
control soil as well as environmental pollution.
6. Earthworms effectively harness the beneficial soil micro flora, destroy soil pathogen and convert
organic wastes in to vitamins, enzymes, antibiotics, PGH, protein rich products and others organic
compounds. More information is needed in this field.
7. Offers new perspectives on vermicomposting research.
CONCLUSION
“Nobody and nothing can be compared with earthworms in their positive influence on the whole
living Nature. They create soil and everything that lives in it. They are the most numerous animals on
Earth and the main creatures converting all organic matter into soil humus providing soil’s fertility
and biosphere’s functions: disinfecting, neutralizing, protective and productive.”
(Anatoly M. Igonin21, Ph. D., Professor at the Vladimir Pedagogical University, Vladimir, Russia, as
quoted in Casting Call 9(2), Aug 2004.)
Aristotle called worms the “intestines of the earth” and Charles Darwin wrote a book on worms and
their activities, in which he stated that there may not be any other creature that has played so
important a role in the history of life on earth (Bogdanov
36
)(Pl. delete). There can be little doubt that
humankind’s relationship with worms is vital and needs to be nurtured and expanded. The following
sections touch on some of the most important areas in which our natural environment can be
An Outlook … Kalam and Ahmad.
63 J. Chem. Bio. Phy. Sci. Sec. D, November 2016 – January 2017; Vol.7, No.1; 049-065.
preserved and sustained through a partnership with these engines of the soil. Vermicompost
containing higher amount of growth promoting substances, vitamins, and enzymes, which in turn
increased the microbial population and the addition Azospirillum increased the root biomass
production, which resulted in higher production of root exudates increasing the beneficial bacteria,
fungi and actinomycetes population in rhizosphere region and influenced the soil physical, chemical
and biological fertility. Vermicompost spread on farm land will not result in pathogen contamination
of ground or surface waters. Also pasturelands seeded and re-seeded with E. feotida cocoons could
help to prevent water contamination by pathogens, since fresh manure dropped by grazing animals
will be quickly colonized by compost worms.
Climate change is one of the most serious and pressing environmental problems of our time. One of
the principal benefits of vermicomposting occurs through carbon sequestration. This is the process of
locking carbon up in organic matter and organisms within the soil. Soils worldwide have been
gradually depleted of carbon through the use of non-organic farming systems. The consistent
application of compost or vermicompost gradually raises the level of carbon in the soil. Although
carbon is constantly leaving the soil as more is being sequestered, the use of composts can increase
the equilibrium level, effectively removing large amounts of carbon permanently from the
atmosphere. Earthworms have an extremely important role to play in counteracting the loss of
biodiversity. Worms increase the numbers and types of microbes in the soil by creating conditions
under which these creatures can thrive and multiply. The earthworm gut has been described as a little
“bacteria factory”, spewing out many times more microbes than the worm ingests. By adding
vermicompost and cocoons to a farm’s soil, you are enriching that soil’s microbial community
tremendously. This below-ground biodiversity is the basis for increased biodiversity above ground, as
the soil creatures and the plants that they help to grow are the basis of the entire food chain. The
United Nations Environment Program (UNEP) has acknowledged the importance of below-ground
biodiversity as a key to sustainable agriculture, above-ground biodiversity, and the overall economy.
ACKNOWLEDGMENT
Authors are grateful to Department of Microbiology, Bidhannagar college for providing infrastructure
& library and also Department of Environment, Govt. of West Bengal for financial assistance to do
the work.
REFERENCES
1. C.A. Edwards and J.R. Lofty. Biology of Earthworms. London: Chapman and Hall
Ltd. 1972, 283.
2. Ernst, David. The Farmer’s Earthworm Handbook. Lessiter Publications,
Brookfield, Wisconsin. 1995, 112.
3. C.A. Edwards. “The Use of Earthworms in the Breakdown and Management of
Organic Wastes”. In: Edwards, C.A. (Ed) Earthworm Ecology. St. Lucie Press,
Boca Raton. 1998, 327-354.
4. R.E. Gaddie. (Senior) and Donald E. Douglas. Earthworms for Ecology and Profit.
Volume 1: Scientific Earthworm Farming. Bookworm Publishing Company,
California. 1975, 180.
5. Myers, Ruth. The ABCs of the Earthworm Business. Shields Publications, Eagle
Rver, Wisconsin, USA. 1969, 64.
6. R.S. Anderson, H.D. Durst, W.G. Landis. Organofluorophosphate- hydrolyzing
activity in an estuarine clam, Rangia cuneata. Comparative Biochemistry and
Physiology. 1988, C 91, 575-578.
63 J. Chem. Bio. Phy. Sci. Sec. D, November 2016 – January 2017; Vol.7, No.1; 049-065.
preserved and sustained through a partnership with these engines of the soil. Vermicompost
containing higher amount of growth promoting substances, vitamins, and enzymes, which in turn
increased the microbial population and the addition Azospirillum increased the root biomass
production, which resulted in higher production of root exudates increasing the beneficial bacteria,
fungi and actinomycetes population in rhizosphere region and influenced the soil physical, chemical
and biological fertility. Vermicompost spread on farm land will not result in pathogen contamination
of ground or surface waters. Also pasturelands seeded and re-seeded with E. feotida cocoons could
help to prevent water contamination by pathogens, since fresh manure dropped by grazing animals
will be quickly colonized by compost worms.
Climate change is one of the most serious and pressing environmental problems of our time. One of
the principal benefits of vermicomposting occurs through carbon sequestration. This is the process of
locking carbon up in organic matter and organisms within the soil. Soils worldwide have been
gradually depleted of carbon through the use of non-organic farming systems. The consistent
application of compost or vermicompost gradually raises the level of carbon in the soil. Although
carbon is constantly leaving the soil as more is being sequestered, the use of composts can increase
the equilibrium level, effectively removing large amounts of carbon permanently from the
atmosphere. Earthworms have an extremely important role to play in counteracting the loss of
biodiversity. Worms increase the numbers and types of microbes in the soil by creating conditions
under which these creatures can thrive and multiply. The earthworm gut has been described as a little
“bacteria factory”, spewing out many times more microbes than the worm ingests. By adding
vermicompost and cocoons to a farm’s soil, you are enriching that soil’s microbial community
tremendously. This below-ground biodiversity is the basis for increased biodiversity above ground, as
the soil creatures and the plants that they help to grow are the basis of the entire food chain. The
United Nations Environment Program (UNEP) has acknowledged the importance of below-ground
biodiversity as a key to sustainable agriculture, above-ground biodiversity, and the overall economy.
ACKNOWLEDGMENT
Authors are grateful to Department of Microbiology, Bidhannagar college for providing infrastructure
& library and also Department of Environment, Govt. of West Bengal for financial assistance to do
the work.
REFERENCES
1. C.A. Edwards and J.R. Lofty. Biology of Earthworms. London: Chapman and Hall
Ltd. 1972, 283.
2. Ernst, David. The Farmer’s Earthworm Handbook. Lessiter Publications,
Brookfield, Wisconsin. 1995, 112.
3. C.A. Edwards. “The Use of Earthworms in the Breakdown and Management of
Organic Wastes”. In: Edwards, C.A. (Ed) Earthworm Ecology. St. Lucie Press,
Boca Raton. 1998, 327-354.
4. R.E. Gaddie. (Senior) and Donald E. Douglas. Earthworms for Ecology and Profit.
Volume 1: Scientific Earthworm Farming. Bookworm Publishing Company,
California. 1975, 180.
5. Myers, Ruth. The ABCs of the Earthworm Business. Shields Publications, Eagle
Rver, Wisconsin, USA. 1969, 64.
6. R.S. Anderson, H.D. Durst, W.G. Landis. Organofluorophosphate- hydrolyzing
activity in an estuarine clam, Rangia cuneata. Comparative Biochemistry and
Physiology. 1988, C 91, 575-578.
An Outlook … Kalam and Ahmad.
64 J. Chem. Bio. Phy. Sci. Sec. D, November 2016 – January 2017; Vol.7, No.1; 049-065.
7. H. Attaway, J.O. Nelson, A.M. Baya, M.J. Voll, W.E. White, D.J. Grimes, R.R.
Colwell. Bacterial detoxification of diisopropylfluorophosphate. Applied
Environmental Microbiology. 1987, 53, 1685-1689.
8. M.M. Benning, J.M. Kuo, F.M. Raushel, H.M. Holden. Three- dimensional
structure of phosphotriesterase: an enzyme capable of detoxifying
organophosphate nerve agent. Biochemistry. 1994, 33, 15001-15007.
9. M. Bradford. A rapid and sensitive method of the quantitation of microgram
quantities of protein utilizing the principle of protein-dye binding. Analytical
Biochemistry. 1976, 72, 248-254.
10. S.R. Caldwell, J.R. Newcomb, K.A. Schlecht, F.M. Raushel. Limits diLusion in
the hydrolysis of substrates by the phosphotriesterase from Pseudomonas
diminuta. Biochemistry. 1991a, 30, 7438-7444.
11. S.R. Caldwell, F.M. Raushel, P.M. Weiss, W.W. Cleland. Transition-state
structures for enzymatic and alkaline phosphotriesterase hydrolysis. Biochemistry.
1991b, 30, 7444-7450.
12. G. Carro-Ciampi, D. Kadar, W. Kalow. Distribution of serum paraoxon
hydrolyzing activities in a Canadian population. Canadian Journal of
Physiological Pharmacology. 1981, 59, 904-907.
13. D.P. Dumas, S.R. Caldwell, J.R. Wild, F.M. Raushel. Purication and properties of
the phosphotriesterase from Pseudomonas diminuta. Journal of Biological
Chemistry. 1989, 264, 19659-19665.
14. H.W. Eckerson, J. Romson, C. Wyte, B. La Du. The human serum paraoxonase
polymorphism: identication of phenotypes by their response to salts. American
Journal of Human Genetics. 1983, 35, 214-227.
15. F. Gil, M.C. Gonzalvo, A.F. Hernandez, E. Villanueva. Rat liver paraoxonase:
subcellular distribution and characterization. Chemico-Biological Interactions.
1993, 87, 149-154.
16. L.L. Harper, C.S. McDaniel, C.E. Miller, J.R. Wild. Dissimilar plasmids isolated
from Pseudomonas diminuta MG and a Flavobacterium sp. (ATCC27511) contain
identical opd gene. Applied Environmental Microbiology. 1988, 54, 2586-2589.
17. F.C. Hoskin, A.H. Roush. Hydrolysis of nerve gas by squid-type
diisopropylphosphorofluoridate hydrolyzing enzyme on agarose resin. Science.
1982, 215, 1255-1257.
18. W.G. Landis, D.M. Haley, M.V. Haley, D.W. Johnson, H.D. Durst, R.E. Savage
Jr.. Discovery of multiple Organofluorophosphate hydrolyzing activities in the
protozoan Tetrahymena thermophila. Journal of Applied Toxico. 1987, 7, 35-41.
19. P.A. Abbasi, J. Al-Dahmani, F. Sahin, H.A.J. Hoitink and S.A. Miller. Effect of
composts on disease severity and yield in organic and conventionally produced
tomatoes. Plant Disease. 2002, 86, 156-161.
20. G.N. Al-Karaki, R.B. Clark & C.Y. Sullivan. Effects of Phosphorous and Water
Stress Levels on Growth and Phosphorous Uptake of Bean and Sorghum Cultivars.
Journal of Plant Nutrition. 1995, 18 (3), 563-578.
21. M.S. Aulakh & J.W. Doran. Kinetics of nitrification under upland and flooded
soils of varying texture. Communications in Soil Science & Plant Analysis. 1996,
27 (9/10), 2079-2089.
22. F. Binet & P. Trehen. Experimental Microcosm Study of the Role of Lumbricus
Terrestris (Oligochaeta: Lumbricadea) on Nitrogen Dynamics in Cultivated Soils.
Soil Biology & Biochemistry. 1992, 13 (1), 39-42.
64 J. Chem. Bio. Phy. Sci. Sec. D, November 2016 – January 2017; Vol.7, No.1; 049-065.
7. H. Attaway, J.O. Nelson, A.M. Baya, M.J. Voll, W.E. White, D.J. Grimes, R.R.
Colwell. Bacterial detoxification of diisopropylfluorophosphate. Applied
Environmental Microbiology. 1987, 53, 1685-1689.
8. M.M. Benning, J.M. Kuo, F.M. Raushel, H.M. Holden. Three- dimensional
structure of phosphotriesterase: an enzyme capable of detoxifying
organophosphate nerve agent. Biochemistry. 1994, 33, 15001-15007.
9. M. Bradford. A rapid and sensitive method of the quantitation of microgram
quantities of protein utilizing the principle of protein-dye binding. Analytical
Biochemistry. 1976, 72, 248-254.
10. S.R. Caldwell, J.R. Newcomb, K.A. Schlecht, F.M. Raushel. Limits diLusion in
the hydrolysis of substrates by the phosphotriesterase from Pseudomonas
diminuta. Biochemistry. 1991a, 30, 7438-7444.
11. S.R. Caldwell, F.M. Raushel, P.M. Weiss, W.W. Cleland. Transition-state
structures for enzymatic and alkaline phosphotriesterase hydrolysis. Biochemistry.
1991b, 30, 7444-7450.
12. G. Carro-Ciampi, D. Kadar, W. Kalow. Distribution of serum paraoxon
hydrolyzing activities in a Canadian population. Canadian Journal of
Physiological Pharmacology. 1981, 59, 904-907.
13. D.P. Dumas, S.R. Caldwell, J.R. Wild, F.M. Raushel. Purication and properties of
the phosphotriesterase from Pseudomonas diminuta. Journal of Biological
Chemistry. 1989, 264, 19659-19665.
14. H.W. Eckerson, J. Romson, C. Wyte, B. La Du. The human serum paraoxonase
polymorphism: identication of phenotypes by their response to salts. American
Journal of Human Genetics. 1983, 35, 214-227.
15. F. Gil, M.C. Gonzalvo, A.F. Hernandez, E. Villanueva. Rat liver paraoxonase:
subcellular distribution and characterization. Chemico-Biological Interactions.
1993, 87, 149-154.
16. L.L. Harper, C.S. McDaniel, C.E. Miller, J.R. Wild. Dissimilar plasmids isolated
from Pseudomonas diminuta MG and a Flavobacterium sp. (ATCC27511) contain
identical opd gene. Applied Environmental Microbiology. 1988, 54, 2586-2589.
17. F.C. Hoskin, A.H. Roush. Hydrolysis of nerve gas by squid-type
diisopropylphosphorofluoridate hydrolyzing enzyme on agarose resin. Science.
1982, 215, 1255-1257.
18. W.G. Landis, D.M. Haley, M.V. Haley, D.W. Johnson, H.D. Durst, R.E. Savage
Jr.. Discovery of multiple Organofluorophosphate hydrolyzing activities in the
protozoan Tetrahymena thermophila. Journal of Applied Toxico. 1987, 7, 35-41.
19. P.A. Abbasi, J. Al-Dahmani, F. Sahin, H.A.J. Hoitink and S.A. Miller. Effect of
composts on disease severity and yield in organic and conventionally produced
tomatoes. Plant Disease. 2002, 86, 156-161.
20. G.N. Al-Karaki, R.B. Clark & C.Y. Sullivan. Effects of Phosphorous and Water
Stress Levels on Growth and Phosphorous Uptake of Bean and Sorghum Cultivars.
Journal of Plant Nutrition. 1995, 18 (3), 563-578.
21. M.S. Aulakh & J.W. Doran. Kinetics of nitrification under upland and flooded
soils of varying texture. Communications in Soil Science & Plant Analysis. 1996,
27 (9/10), 2079-2089.
22. F. Binet & P. Trehen. Experimental Microcosm Study of the Role of Lumbricus
Terrestris (Oligochaeta: Lumbricadea) on Nitrogen Dynamics in Cultivated Soils.
Soil Biology & Biochemistry. 1992, 13 (1), 39-42.
An Outlook … Kalam and Ahmad.
65 J. Chem. Bio. Phy. Sci. Sec. D, November 2016 – January 2017; Vol.7, No.1; 049-065.
23. M.J. Boehm, L.V. Madden, H.A.J. Hoitink. Effect of organic matter
decomposition level on bacterial species diversity and decomposition in
relationship to Pythium damping-off severity. Applied and Environmental
Microbiology. 1993, 59, 4171-4179.
24. I.H. Chaoui, L.M. Zibilske, S. Ohno. Effect of earthworm casts and compost on
microbial activity and plant nutrient uptake. Soil Biol. And Biochem. 2003, 35,
295-302.
25. O. Daniel, J.M. Anderson. Microbial Biomass and Activity in Contrasting Soil
Materials after Passage through the Gut of the Earthworm Lumbricus Rubellus
Hoffmeister. Soil Biology & Biochemistry. 1992, 24 (5), 465-470.
26. T. Decaens, A.F. Rangel, N. Asakawa, R.J. Thomas. Carbon and nitrogen
dynamics in ageing earthworm casts in grasslands of the eastern plains of
Colombia. Biology and Fertility of Soils. 1999, 30 (1-2), 20-28.
27. W. Devliegher & W. Verstraete. The effect of Lumbricus terrestris on soil in
relation to plant growth: effects of nutrient-enrichment processes (NEP) and gut-
associated processes (GAP). Soil-Biology-and-Biochem. 1997, 29 (3/4), 341-346.
28. S.C. Park, T.J. Smith, M.S. Bisesi. Bioactivation of bis (p nitro phenyl) phosphate
by phosphotriesterase of the earthworm. Lumbricus terrestris. Drug and Chemical
Toxicology. 1993, 16, 111-116.
29. B.L. Roberts, H.W. Dorough. Relative toxicities of chemicals to the earthworm
Eisenia andrei. Environmental Toxicology and Chemistry. 1984, 3, 67-78.
30. H. Becker, P.J. Edwards, F. Heimbach & P.W. Greig-Smith. Eeotoxicology of
Earthworms. Intercept Ltd., Hampshire, UK. 1992.
31. G. Bengtsson, T. Gunnarsson & S. Rungren. Effects of metal pollution on the
earthworm Dendrobaena ruhida (Sav) in acidified soils. Water Air Soil Pollut.
1986, 28, 361-83.
32. R. Dallinger & P.S. Rainbow. Ecotoxicology of Metals" in Terrestrial
Invertebrates. Lewis Publishers, Chelsea, USA. 1993.
33. EEC. EEC Directive 79/831. Annex V. Part C. Methods for the determination of
ecotoxicity. Level I. C (II) 4: Toxicity for earthworms. Artificial soil test.
DGXI/128/82. 1985.
34. C.A. Gestel, M. van & W.A. Dis, A. van. The influence of soil characteristics on
the toxicity of four chemicals to the earthworm Eisenia andrei (Oligochaeta). Biol.
Fertil. Soils. 1988, 6, 262-5.
35. C.A. Gestel, M. van, W.A. Dis, A. van, E.M. Breemen van & P.M. Sparenburg.
Development of a standardized reproduction toxicity test with the earthworm
species Eisenia andrei using copper, pentachlorophenol and 2.4- dichloroaniline.
Ecotoxicol. Environ Safety. 1989, 18, 305, 12.
*Corresponding author: Abul Kalam; Department of Microbiology,
Bidhannagar Govt College, Salt Lake City, Kolkata, India. [email protected]
65 J. Chem. Bio. Phy. Sci. Sec. D, November 2016 – January 2017; Vol.7, No.1; 049-065.
23. M.J. Boehm, L.V. Madden, H.A.J. Hoitink. Effect of organic matter
decomposition level on bacterial species diversity and decomposition in
relationship to Pythium damping-off severity. Applied and Environmental
Microbiology. 1993, 59, 4171-4179.
24. I.H. Chaoui, L.M. Zibilske, S. Ohno. Effect of earthworm casts and compost on
microbial activity and plant nutrient uptake. Soil Biol. And Biochem. 2003, 35,
295-302.
25. O. Daniel, J.M. Anderson. Microbial Biomass and Activity in Contrasting Soil
Materials after Passage through the Gut of the Earthworm Lumbricus Rubellus
Hoffmeister. Soil Biology & Biochemistry. 1992, 24 (5), 465-470.
26. T. Decaens, A.F. Rangel, N. Asakawa, R.J. Thomas. Carbon and nitrogen
dynamics in ageing earthworm casts in grasslands of the eastern plains of
Colombia. Biology and Fertility of Soils. 1999, 30 (1-2), 20-28.
27. W. Devliegher & W. Verstraete. The effect of Lumbricus terrestris on soil in
relation to plant growth: effects of nutrient-enrichment processes (NEP) and gut-
associated processes (GAP). Soil-Biology-and-Biochem. 1997, 29 (3/4), 341-346.
28. S.C. Park, T.J. Smith, M.S. Bisesi. Bioactivation of bis (p nitro phenyl) phosphate
by phosphotriesterase of the earthworm. Lumbricus terrestris. Drug and Chemical
Toxicology. 1993, 16, 111-116.
29. B.L. Roberts, H.W. Dorough. Relative toxicities of chemicals to the earthworm
Eisenia andrei. Environmental Toxicology and Chemistry. 1984, 3, 67-78.
30. H. Becker, P.J. Edwards, F. Heimbach & P.W. Greig-Smith. Eeotoxicology of
Earthworms. Intercept Ltd., Hampshire, UK. 1992.
31. G. Bengtsson, T. Gunnarsson & S. Rungren. Effects of metal pollution on the
earthworm Dendrobaena ruhida (Sav) in acidified soils. Water Air Soil Pollut.
1986, 28, 361-83.
32. R. Dallinger & P.S. Rainbow. Ecotoxicology of Metals" in Terrestrial
Invertebrates. Lewis Publishers, Chelsea, USA. 1993.
33. EEC. EEC Directive 79/831. Annex V. Part C. Methods for the determination of
ecotoxicity. Level I. C (II) 4: Toxicity for earthworms. Artificial soil test.
DGXI/128/82. 1985.
34. C.A. Gestel, M. van & W.A. Dis, A. van. The influence of soil characteristics on
the toxicity of four chemicals to the earthworm Eisenia andrei (Oligochaeta). Biol.
Fertil. Soils. 1988, 6, 262-5.
35. C.A. Gestel, M. van, W.A. Dis, A. van, E.M. Breemen van & P.M. Sparenburg.
Development of a standardized reproduction toxicity test with the earthworm
species Eisenia andrei using copper, pentachlorophenol and 2.4- dichloroaniline.
Ecotoxicol. Environ Safety. 1989, 18, 305, 12.
*Corresponding author: Abul Kalam; Department of Microbiology,
Bidhannagar Govt College, Salt Lake City, Kolkata, India. [email protected]
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