Microbial activity in Vermicompost and Vermiwash Review

2. Microbial diversity of
vermicompost and vermiwash
and their significance in agriculture
Speaker : Roshan Nisarta
Degree : M. Sc. (Agri.) Agril. Microbiology
Major Advisor : Mrs. H. N. Shelat
Minor Advisor : Dr. S. N. Shah
Course No. : MICRO. 591
Reg. No. : 04-2874-2016
Date : 26/12/2017
Time : 2:30 p.m.
Seminar On

3. CONTENT
• INTRODUCTION
• WHAT IS VERMICOMPOST AND VERMIWASH
• PREPARATION FROM WASTE
• VARIOUS MICROFLORA -NUMBER AND DIVERSITY
• ROLE OF MICROFLORA - ENZYMES AND BIOCHEMCIAL REACTIONS
• CASE STUDIES
• CONCLUSION
• FUTURE THRUST
2

4. Introduction
• Vermicomposting is a non-thermophilic, bio oxidative process that involves
earthworms and associated microbes that converts organic debris into
worm castings.
• Vermicompost is a finely divided, peat like material with high porosity, good
aeration, drainage, water holding capacity, microbial activity, excellent
nutrient status and buffering capacity thereby resulting in the required
physiochemical characters congenial for soil fertility and plant growth.
3

5. • Vermicomposting systems sustain a complex microbial and invertebrate food web that
results in the recycling of organic matter and release of nutrients
• Earthworms are important drivers of soil biogeochemical processes as they modify soil
physicochemical properties and microbial communities by feeding, burrowing and casting
activities
• The proximate activities of earthworms enhance the mineralization of both carbon and
nitrogen in the substrate significantly and such effects are in proportion to the earthworm
population densities
• Although microorganisms are mainly responsible for the biochemical degradation of
organic matter, earthworms play an important role in the process by fragmenting and
conditioning the substrate, increasing surface area for growth of microorganisms and
altering its biological activity
4

6. • During Vermicomposting process, when organic matter passes through the earthworm gut, it
undergoes physical, chemical and biochemical changes by the combined effect of earthworm and
microbial enzymatic activities.
• The effect of earthworms on the decomposition of organic waste is due to gut-associated processes
(GAPs). These processes include all the modifications including
1. The addition of sugars and other substances
2. Modification of the microbial diversity and activity
3. Homogenization and the intrinsic processes of digestion
4. Assimilation and production of mucus and excretory substances such as urea and ammonia,
which constitute a readily assimilable pool of nutrients for microorganisms.
5

7. • Vermicompost enhances soil biodiversity by promoting the beneficial microbes population
which are the most numerically abundant and diverse members of the vermicomposting food
web
• The role of microbes in the gut (cellulase produced by the fungi plays a major role in
decomposition of cellulolytic materials of organic wastes) as well as in the cast is very
essential for the degradation of organic wastes and release of nutrients to plants.
• Endosymbiotic microbes produce extracellular enzymes that degrade cellulose and phenolic
compounds, enhancing the degradation of ingested material and the degraded organic
matter passes out of the earthworm’s body in the form of casts.
• Microbial biomass in the worm casts was found to be high and their activity was essential for
release of nutrients into the medium so that it can be taken by the plants.
6

8. • The primary consumers of the vermicomposting food web are the
microorganisms that break down and mineralize organic residues
 Microbes form a part of food for earthworm
 Microbes are proliferated in the gut and vermicompost
 Earthworm help in the distribution of microbes
 Together with earthworm microbes mineralize, humifies organic matter,
facilitates chelation etc.
7

9. Methods for vermicomposting
Bed method:
Composting on the pucca / kachcha floor by
making bed (6x2x2 feet size) of organic mixture.
This method is easy to maintain and to practice
Pit method:
Composting in the cemented pits of size 5x5x3
feet. Unit is covered with thatch grass or any
other locally available materials. This method is
not preferred due to poor aeration, water
logging at bottom, and more cost of production
8

10. Waste materials for
Vermicompost
Crop
residues
Weed
biomass
Waste
from
Agro-
industries
Urban &
Rural
wastes
Leaf litter
Vegetable
waste
Hotel
refuse
9

11. Steps followed for vermicompost preparation
Cow dung
and
chopped
dried leafy
(3:1)
Partial
decomposition
for 15-20 days
A layer of
15-20 cm
of chopped
dried
leaves/
grasses as
bedding
material at
the bottom
of the bed.
Beds of
partially
decomposed
material of
size 6 x 2 x 2
feet should
be made
Each bed
should
contain
1.5-2.0 q
of raw
material
Red
earthworm
(1500-
2000)
should be
released
on the
upper
layer of
bed.
Water
sprinkling
with
immediately
after the
release of
worms.
Maintain
moisture
and
humidity
and turned
once after
30 Days
Compost
gets ready
in 45-50
days
10

12. • Collection of wastes, shredding, mechanical separation of the metal, glass, ceramics
and storage of organic wastes
Phase 1:
• Pre digestion of organic waste for twenty days by heaping the material along with
cattle dung slurry make them fit for earthworm consumption.
Phase 2:
• Preparation of earthworm bed
Phase 3:
• Collection of earthworm after vermicompost harvest. Sieving the composted
material to separate fully composted material. The partially composted material will
be again put into vermicompost bed
Phase 4:
• Storing the vermicompost in proper place to maintain moisture and allow the
beneficial microorganisms to grow.
Phase 5:
Phase of vermicomposting
11

13. Advantages of Vermicompost
Free flowing, easy
to apply, handle
and store and does
not have bad odour
Rich in all essential
plant macro and
micro nutrients
Improves physical
properties
Improves
biological
properties &
activities in soil
Phytotonic effect
and increase yield
and product quality
Increases the
population and
activity of
earthworm in the
soil
Increases the use
efficiency of
chemical fertilizers
Eliminate weed
seeds and
Pathogens
Improve immune
system of plant
Enhances the
decomposition of
organic matter in
soil
Contains valuable
vitamins, enzymes
and hormones like
auxins, gibberellins
etc.
Improves stress
tolerance in plants
Keep the plants
lush green
12

14. • Vermiwash is a liquid leachate collected by allowing excess water to saturate the actively
vermicomposting substrate in such a way that the water washes the nutrients from the vermicast excreted
by the earthworms feeding on the substrate as well as the earthworm’s body surface.
• It’s a liquid fertilizer for foliar spray, collected by passing water in a regulated way through a column of
active live local earthworms being cultured in a container, being fed with cattle dung and straw.
Vermiwash
A barrel
(250L) or
even a
small
bucket
Broken
stones
Coarse
sand
Garden
soil
Earth
worms
Cattle
dung
Straw Water
Materials Needed
13

15. Collect the watery yellowish to black extract of vermicompost, vermiwash drainage out off drum
Leave 15 days to acclimatize
Moisten the soil well and offer them cattle dung and straw as feed
Fill with vermicompost with heavy population of earth worms
Make a layer of sand (2-3 cm) thickness
Fill the first layers with broken bricks, about 10 cm thickened as shown above
Set up the drum or barrel with an outlet on a couple of bricks to facilitate collection of vermiwash
Preparation Method
14

16. Contains nitrogen
fixing bacteria and
Phosphate
solubilizing bacteria
Contains plant
growth hormones
like auxins and
cytokinin
Acts as a plant tonic
and helps to reduce
many plant diseases
Act as biopesticide
and liquid manure
It is an organic
growth and immunity
booster in plants
It contains high
microbes content
which prevents
plants from
infections
Rich in NPK, amino
acids and other
micro nutrient
required by the
plants
Complete food for
plants which
enhances the quality
of the produce
Vermiwash Benefits
15

17. Table 1. Physico-chemical properties of vermiwash, vermicompost and soil
Punjab (India) Makkar et al. (2017)
Parameter
Vermiwash
(mg/L)
Vermicompost
(g/kg)
Soil
(g/kg)
pH 6.12 7.11 8.6
N 0.5 2.5 0.12
P 0.8 8.4 0.9
K 1.2 8.0 4.2
mg/kg mg/kg
Ca 130.0 240 149.9
Mn 0.01 0.09 0.008
Cu 0.04 5.0 0.87
Zn 0.9 24.5 1.1
16

18. Table 2. Earthworm species used in vermicomposting
Ecological
category
Species Habitats Family
Epigeic • Dichogaster
Bolaui,
• Perionyx
simlaensis
Sewage,
Irrigation
channels
Octochaetidae
Megascolecidae
Endogeic • Drawida
nepalensis,
• Octochaeton
a beatrix
Garden ,
Nurseries,
Cultivated
fields
Monoligastridae
Octochaetidae
Epi-anecic • Metaphire
posthuma
Cultivated
fields,
Sewage,
Grassland,
Garden, Non-
Cultivated
areas
Megascolecidae
D. bolaui P. simlaensis D. nepalensis
O. beatrix M. posthuma
17

19. Eudrilus eugeniae Eisenia foetida
Perionyx excavatus
18

20. Table 3. Ecological categories and niches of earthworms and their characteristic features and beneficial traits
Species
Ecological
category
Ecological niche Characteristic features Beneficial trait
E. foetida, E. eugeniae,
Lumbricus rubellus,
L. castaneus, L. festivus
Eiseniella tetraedra,
Bimastus minusculus,
B. eiseni, Dendrodrilus rubidus,
Dendrobaena veneta, D. Octaedra
Epigeics
Superficial soil
layers,
leaf litter,
compost
Smaller in size, body uniformly pigmented,
active gizzard, short life cycle, high
reproduction rate and regeneration, tolerant to
disturbance, phytophagous
Efficient bio-degraders and nutrient
releasers, efficient compost producers, aids
in litter comminution and early
decomposition
Aporrectodea caliginosa Endogeics
Topsoil or
subsoil
Small to large sized worms, weakly
pigmented, life cycle of medium duration,
moderately tolerant to disturbance,
geophagous
Brings about pronounced changes in soil
physical structure, can efficiently utilize
energy from poor soils hence can be used
for soil improvements
Octolasion cyaneum
O. lacteum,
Polyhumic
endogeic
Top soil (A1)
Small size, unpigmented, forms horizontal
burrows, rich soil feeder
Pontoscolex corethrurus
Allolobophora chlorotica
Mesohumi
c
endogeic
A and B horizon
Medium size, unpigmented, forms extensive
horizontal burrows, bulk (A1) soil feeder
Aminthas sp.
Oligohumi
c
endogeic
B and C horizon
Very large in size, unpigmented, forms
extensive horizontal burrows, feeds on poor,
deep soils
L. terrestris,
L. Polyphemus
A. longa
Anecics
Permanent deep
burrows in soil
Large in size, dorsally pigmented, forms
extensive, deep, vertical permanent burrows,
low reproductive rate, sensitive to
disturbance, phytogeophagous, Nocturnal
Forms vertical burrows affecting air-water
relationship and movement from deep
layers to surface helps in efficient mixing
of nutrients
19

21. Microbial actions in Vermicompost and Vermiwash
Action of
Microbes
Production of
enzymes
Production of
acids
Production of
growth
promoting
substances
Antibiotic
production
20

22. Table 4. Microbes (Bacteria and Fungi) diversity in different Vermicompost
systems
Different
Compost
Bacteria Fungal
EE/CA
Enterobactoer sp., Pseudomonas sp., Flavobacteriam,
Cytophaga sp., Staphylococcus aureus
Zygosaccharomyces sp.,
Saccharomyces sp., Aspergilus flavus
EE/TP
Proteus vulgaris, Azotobacter sp., Aeromonas sp,
Pseudomonas sp, S. aureus, E.coli
A. flavus, Fusarium roseu,
Mucor variens, A. niger, A. fumigates
EF/CA
Gordonia sp., Micrococcus sp., Cytophaga sp., Aeromonas sp,
Cellulomonas sp, S. aureus
Rhizopus sp., Candida sp., A. flavus, F. roseu
EF/TP
Cytophaga sp., Aeromonas sp, Cellulomonas sp,
Pseudomonas sp, Staphylococcus aureus
A. flavus, F. roseu, A. niger,
A. fumigates, Geotrichum candidum
Anonymous, (2012)
EE/CA – E. eugeniae treated Cassia auriculata compost
EE/TP – E. eugeniae treated Tephrosia purpurea compost
EF/CA – E. foetida treated C. auriculata compost
EF/TP – E. foetida treated T. purpurea compost
21

23. Table 5. Biodiversity of vermicompost bacteria and their beneficial traits
Vermicompost
earthworm
Names of bacteria Beneficial traits References
Pheretima sp. P. oxalaticus Oxalate degradation Khambata and Bhat, 1953
Unspecified Rhizobium trifolii Nitrogen fixation and growth of leguminous plants Buckalew et al. 1982
Lumbricus rubellus R. japonicum, P. putida Plant growth promotion Madsen and Alexander 1982
L. Terrestris Bradyrhizobium japonicum Improved distribution of nodules on soybean roots Rouelle, 1983
Aporrectodea
trapezoids
A. Rosea
P. corrugata 214OR Suppress Gaeumannomyces graminis var. Tritd in wheat Doube et al. 1994
A. trapezoids R. meliloti L5-30R Increased root nodulation and nitrogen fixation in legumes Stephens et al. 1994b
Eisenia foetida Bacillus spp., B. megaterium, B. pumilus, B.
subtilis, Actinobacteria
Antimicrobial activity against Enterococcus faecalis DSM
2570, Staphylococcus aureus DSM 1104
Antifungal activity against Colletotrichum coccodes,
R. solani, P. ultimum, P. capsici and F. moliniforme
Moreira et al. 2008
Yasir et al. 2009
Lumbricus terrestris Fluorescent pseudomonads, Filamentous
Actinomycetes
Suppress Fusarium oxysporum f. sp. asparagi and F.
proliferatum in asparagus, Verticillium dahlia in eggplant
and F. oxysporum f. sp. lycopersici Race 1 in tomato
Elmer, 2009
Eudrilus sp. Free-living N2 fixers, Azospirillum,
Azotobacter, Autotrophic Nitrosomonas,
Nitrobacter, Ammonifyingbacteria, Phosphate
solubilizers, Fluorescent pseudomonads
Plant growth promotion by nitrification, phosphate
solubilisation and plant
disease suppression
Gopal et al. 2009
Unspecified Eiseniicola composti YC06271T Antagonistic activity against F. moniliforme Yasir et al. 2009b
22

24. Fig. 2. Number of bacteria and
actinomycetes in the various
sections of the gut of Lumbricus
terrestris (Linn.)
0
2
4
6
8
10
12
14
Fore-gut Mid-gut Hind-gut
Log.no.
Actinomycetes
Bacteria
Fig. 1. Diagram of Earthworm gut
23

26. Case study: 1
Studied the diversity and quantitative study of bacteria from
vermiwash samples on Nutrient agar medium, Jenson’s medium and
CRYMA medium
Maharashtra, India Patil et al. (2014)
Treatment Detail
VM-1 Vermiwash Prepared with Horse dung
VM-2 Vermiwash prepared with pig dung
VM-3 Vermiwash prepared with elephant dung
VM-4 Vermiwash prepared with cow dung
25

27. Table 6. Bacterial population from different vermiwash samples on various bacterial media
Sample
SPC/ ml of bacteria on
Nutrient agar medium
SPC/ ml of bacteria on
Jenson’s medium
SPC/ ml of bacteria on
CRYMA medium
VM-1 42 X 103 34 X 103 38 X 102
VM-2 30 X 101 32 X 101 33 X 101
VM-3 76 X 101 29 X 102 36 X 101
VM-4 40 X 102 54 X 102 44 X 103
Table 6. The bacterial diversity of various vermiwash samples
Sample Growth of Microorganism
VM-1 P. aeroginosa
VM-2 Rhizobium spp. and Azotobacter spp.
VM-3 P. aeroginosa
VM-4 Azotobactor spp. and Rhizobium spp.
26

28. Case study: 2
Investigated the microbial diversity of vermicompost and vermiwash
prepared by E. euginae from different leaf litter waste and analyzed the bacterial,
fungal and actinomycetes population at initial and 60th day of vermicomposting (24
& 48 hours of extraction of vermiwash)
Esakkiamal et al. (2015)Tamilnadu, India.
Treatment detail
C Cow dung
E1 Mangifera indica leaf waste
E2 Syzygium cumini leaf waste
E3 Vigna radiata leaf waste
E4 Vigna mungo leaf waste
27

29. Total Bacterial, Fungal and Actinomycetes population of E. eugeniae treated
with different leaf litter vermicompost
0
500
1000
Control E1 E2 E3 E4
BacterialPopulation
108/g
Treatments
Initial 60th Day
0
100
200
Control E1 E2 E3 E4
Fungalpopulation108/g
Treatments
Initial 60th Day
Fig. 4. Total Fungal population of E. eugeniae treated with
different leaf litter vermicompost
Fig. 3. Total Bacterial population of E. eugeniae treated
with different leaf litter vermicompost
0
20
40
Control E1 E2 E3 E4
Actinomycetes
population104/g
Treatments
Initial 60th Day
Fig. 5. Total Actinomycetes count of E. eugeniae treated with
different leaf litter vermicompost 28

30. Total Bacterial, Fungal and Actinomycetes count of E. eugeniae treated with
different leaf litter vermiwash
0
200
400
600
Compost E1 E2 E3 E4
Bacterialpopulation
108/g
Treatments
24 Hours 48 Hours
Fig. 6. Total Bacterial population of E. eugeniae treated with
different leaf litter vermiwash
0
20
40
60
80
Compost E1 E2 E3 E4
Fungalpopulation
105/g
Treatments
24 Hours 48 Hours
Fig. 7. Total Fungal population of E. eugeniae treated with
different leaf litter vermiwash
0
10
20
30
Compost E1 E2 E3 E4
Actinomycetes
population104/g
Treatments
24 Hours 48 Hours
Fig. 8. Total Actinomycetes population of E. eugeniae treated with different
leaf litter vermiwash
29

31. Case study: 3
Studied the microbial population and their activity on vermicompost
by E. eugeniae and E. foetida in different concentrations of 400, 500 and 600
g tea waste (TW) with cow dung(CD) and kitchen waste(KW)
Emperor and Kumar (2015)Tamilnadu, India
30

32. Table. 8 Total microbial population in different treatment of tea waste + cow dung + kitchen waste
mixture and vermicompost with E. eugeniae and E. foetida
Sr. No.
Total Microbial Population CFU x 106 g ¹
Treatment
Initial
Substrate
Natural
compost
E. eugeniae E. foetida
1 T1 control soil (1000g soil) 2.87 3.10 3.38 3.31
2 T2 (400g TW + 200g CD + 400g KW) 3.10 3.28 3.56 3.51
3 T3 (500g TW + 100g CD + 400g KW) 3.23 3.47 3.67 3.62
4 T4 (600g TW + 100g CD + 300g KW) 3.32 3.58 3.89 3.84
Treatment Initial Natural Compost
Vermicompost
E. eugenia E. foetida
T1 3.11±0.16 4.17±0.65 5.72±0.61 4.50±0.11
T2 3.21±0.17 4.27±0.67 6.12±0.27 5.46±0.29
T3 3.27±0.23 4.46±0.28 6.60±0.15 5.95±0.63
T4 3.31±0.51 4.82±0.43 7.32±0.31 6.92±0.59
Table 9. Microbial Activity in initial, natural compost, vermicompost produced by two earthworm
species E. eugeniae and E. foetida
31

33. Table 10. Fungal flora isolated from the vermicompost of industrial tea waste, cow
dung and kitchen waste mixtures treated with E. eugeniae and E. foetida
Sr.
No
Fungal Species
Natural
compost
Vermicompost
E.
eugeniae
E.
foetida
1 Aspergilus niger - + +
2
Penicillium
citrinum
- + -
3
Cladosporium
herbarium
+ + -
4
Chaetomium
globossum
+ - +
5 A. nidulans + + -
6 Rhizopus nigricans - + +
7 Mucor plumbers + - -
Fig. 9 Culture of fungus
32

34. Table 11. Bacteria species and Actinomycetes isolated from the vermicompost of industrial tea waste,
cow dung and kitchen waste mixtures with E. eugeniae and E. foetida
Bacterial Species
Natural
Compost
Vermicompost
E. eugeniae E. foetida
Klebsiella pneumoniae (G+ve) - + +
Morganella morgarii (G+ve) - + -
Citrobactor diversus (G+ve) + - +
Enterococcus faecium (G+ve) + + -
Bacillus subtilis (G+ve) - + -
Bacillus cereus (G+ve) - + -
Actinomycetes
Streptomyces albus - + +
Nocardia cariae - + -
Fig. 10. Culture of bacteria
Fig. 11. Culture of Actinomycete
33

35. Case study: 4
Carried out the quantitative estimation of microorganisms in
vermicompost and bio compost by serial dilution method on
nutrient agar medium
Chavan et al. (2013)Thane, India
34

36. Fig. 12. Microbial populations (106 CFU/g) in vermicompost and bio compost
35

37. Case study: 5
• Carried out microbial analysis of vermicompost and gut of
red earthworm for its quantitative study and the existence of
microbes
Yami et al. (2003)Kathmandu, Nepal 36

38. Fig. 13. Distribution of different micro-
organisms in vermicompost
Fig. 14. Distribution of different micro-
organisms in vermicast
Bacteria
68%
Actinomycetes
14%
Fungi
18%
Bacteria
Actinomycetes
Fungi
Bacteria
71%
Actinomycetes
9%
Fungi, 20%
Bacteria
Actinomycetes
Fungi
Bacteria,
73%
Actinomycetes,
10%
Fungi,
17%
Bacteria
Actinomycetes
Fungi
Fig. 15. Distribution of different
micro-organisms in gut
of earthworm
Distribution of different Microorganisms
37

39. Fig. 16. Distribution of different micro-
organisms in vermicompost
Bacillus spp., 39%
Pseudomonas spp.,
29%
Azotobacter
spp., 23%
Others, 9%
Bacillus spp.
Pseudomonas spp.
Azotobacter spp.
Others
Others
12%
Azotobacter spp. 7%
Bacilus spp.
44%
Pseudomon
as spp.
37%
Others
Azotobacter spp
Bacilus spp
Pseudomonas spp
Others
15%
Bacilus spp. 45%
Azotobacter spp. 10%
Pseudomona
s spp. 30%
Others
Bacilus spp
Azotobacter spp
Pseudomonas spp
Fig. 17. Distribution of different
bacteria in vermicast.
Fig. 18. Distribution of different
bacteria in gut of earthworm.
Distribution of Bacteria
38

40. Others 25%
Aspergilus
spp.
50%
Mucor spp.
17%
Rizopus spp.
8%
Others
Aspergilus spp
Mucor spp
Rizopus spp
Fig. 19. Distribution of different
fungi in vermicompost.
Fig. 20. Distribution of different fungi in
vermicast
Others 17%
Aspergilus
spp. 48%
Fusarium
spp. 29%
Curvularia
spp. 6%
Others
Aspergilus spp.
Fusarium spp.
Curvularia spp.
Aspergillus spp.
48%
Curvularia spp.
20%
Fusarium spp.
8%
Geotrichum spp.
16%
Others
8%
Aspergillus spp.
Curvularia spp.
Fusarium spp.
Geotrichum spp.
Fig. 21. Distribution of different fungi in
gut of earthworm
Distribution of Fungi
39

41. Micromonospora spp.
63%
Norcardi
a spp.
31%
Streptomyces spp.
6%
Micromonospora spp.
Norcardia spp.
Streptomyces spp.
Fig. 24. Distribution of different
actinomycetes in gut of
earthworm.
Streptomyces spp.
12%
Norcardia
spp
31%
Micromono
spora spp.
57%
Streptomyces spp
Norcardia spp
Micromonospora spp
Fig. 22. Distribution of different
actinomycetes in vermicompost
Micromonospora spp.
35%
Streptomyces spp.
12%
Nocardia
spp,
53%
Micromonospora spp.
Streptomyces spp.
Nocardia spp
Fig. 23. Distribution of different
actinomycetes in Vermicast
Distribution of Actinomycetes
40

42. Fig. 25. Distribution of microflora in vermicompost, vermicast and gut of E. foetida
41
0
20
40
60
80
100
120
140
160
Vermicompost Gut Cast
Population
No. of bacteria No. of fungi No. of Actinomycetes

43. Case study: 6
• Carried out microbial study of different types of Vermicompost with
control at initial and over 45 days with different species of earthworm
Anonymous, (2012)Itzatnagar, India 42

44. Fig. 26. Total Bacterial count observed in control and different types of
Vermicompost over 45 days
EE/CA - Eudrilus eugeniae treated
Cassia auriculata compost
EE/TP – Eudrilus eugeniae treated
Tephrosia purpurea compost
EF/CA – Eisenia foetida treated
Cassia auriculata compost
EF/TP - Eisenia foetida treated
T. purpurea compost
43

45. Case study: 7
• Studied the digestive enzymes in the gut of earthworms E. euginae
and E. foetida for the quantitative analysis of amylase, cellulase,
xylanase, cellobiase, endoglucanase, acid phosphatase, alkaline
phosphatase and nitrate reductase in different amounts in both species
Prabha et al. (2007)Coimbatore, India
44

46. Fig. 27. Activity of enzymes in the gut of E. eugeniae and E.
foetida. Unit: Amylase, mg of maltose/min/mg of
starch; Cellulose (mg of glucose/min/mg protein;
Xylanase, mmoles/mg protein; Cellobiase, µmol/mg
protein; and Endoglucanase µ moles/mg protein)
Fig. 28. Activity of enzymes in the gut of E. eugeniae and E.
foetida. Unit: Acid phosphatase, mg of phosphorus/min/mg
protein; alkaline phosphatase (mg of phosphorous/min/mg
protein; and Nitrate reductase, µmoles of nitrate/min/mg
protein)
45

47. Case study: 8
Carried out a comparative assessment of enzyme activities and
microbial populations during normal composting and vermicomposting
and in vermicomposting process and studied for the quantitative traits
for the enzyme production
Devi et al. (2008)Tirupati, India
46

48. Incubation period (Days)
Microbialpopulation
(CFUg-1)
Fig. 29: Changes in different microbial populations (106 CFU/g)of
normal compost and vermicompost at weekly intervals 47

49. Changes in cellulase, invertase and amylase activities of normal compost and vermicompost
at weekly intervals µg reducing sugar g-1 h-1 compost
0
200
400
600
800
1000
1200
1400
1 7 14 21 28 35 42 49
Normal Compost vermicompost
Incubation period (Days)
Callulase(μgreductingsugar
g-1hr-1)
Enzyme activities during vermicomposting
0
100
200
300
400
500
600
700
800
900
1 7 14 21 28 35 42 49
Normal Compost vermicompost
Amylase(μgreductingsugar
g-1hr-1)
0
100
200
300
400
500
600
700
800
900
1000
1 7 14 21 28 35 42 49
Normal Compost vermicompost
Invertase(μgreductingsugar
g-1hr-1)
Incubation period (Days)Incubation period (Days)
Fig. 30. Changes in Cellulase activity Fig. 31. Changes in Invertase activity Fig. 32. Changes in Amylase activity
48

50. Changes in urease and protease activities of normal compost and vermicompost at weekly
intervals
0
50
100
150
200
250
1 7 14 21 28 35 42 49
Normal Compost vermicompost
Incubation period (Days)
urease(μgreducingsugar
g-1hr-1)
0
5
10
15
20
25
30
1 7 14 21 28 35 42 49
Normal Compost vermicompost
Protease(μgreducingsugar
g-1hr-1)
Incubation period (Days)
Fig. 33. Changes in Urease activity Fig. 34. Changes in Protease activity
49

51. Case study: 9
Studied qualitative and quantitative analysis of vermiwash
Zambare et al. (2008)Ahmednagar, India
50

52. Table 12. Qualitative estimation of enzyme
secretion from earthworms.
Table 13. Quantitative estimation of enzyme
secretion from earthworms.
Enzymes Zone of clearance (mm)
Gelatinase 17.3 ± 2.7
Amylase 1.85 ± 0.5
Urease Positive
Enzymes
Enzyme activity
(U ml-1min-1)
Specific activity
(U mg-1 Protein)
Gelatinase 115.60 ± 17.23 16.50
Amylase 75 ± 25 10.71
Caseinase 48.16 ± 19.27 6.88
Fig. 35. Enzyme profile of Vermiwash – qualitative method
(A- amylase test, B- protease test and C- urease test) 51

53. Case study: 10
Carried out isolation and identification of bacterial cultures from
the intestine of earthworms. They further inoculated these cultures in
cattle manure and saw dust and fed to earthworm and observed the
biomass and growth related parameters of earthworm.
Korea Hong et al. (2011)
52

54. Table 14. Parameters of earthworm biomass after 8 weeks of vermicomposting
in mixtures that were inoculated by microorganisms.
Treatment
Parameters of earthworm biomass
RM (g)
SR1(%) NE FW1 (mg) FW2 (mg) GR (mg/h) NYE NC CW (g) CW (%)
PG2 56.67c 5.67c 753.02a 598.46b -0.23b 0.67b 0.33c 19.9c 31.3a 43.7c
AH 100.00a 10.0a 753.01a 802.29a 0.09ab 4.00a 3.00ab 23.5b 36.1b 41.5b
PM 100.0a 10.00 a 753.02a 898.62a 0.21ª 4.33a 3.33a 25.44a 40.91c 36.78a
Control* 53.33c 5.33c 753.02a 567.10b -0.28b 2.67ab 2.00bc 17.09f 28.82ab 43.21c
 SR, Survival rate (%);
 NE, average number of earthworms;
 FW1, initial fresh weight of adult earthworms (mg);
 FW2, final fresh weight of adult earthworms (mg);
 GR, growth rate of adult earthworms (mg h-1);
 NYE, average number of young earthworms;
 NC, average number of cocoons;
 CW (g), dry weight of earthworm casts (<2.0 mm);
 CW (%), ratios of earthworm casts;
 RM, dry weight of residual matter (>2.0 mm).
 Results are expressed as means. Values that are followed by
different letters are statistically significant (analysis of variance
(ANOVA), P < 0.05).
 PG, Photobacterium ganghwense
 AH Aeromonas hydrophila
 PM, Paenibacillus motobuensis
 Food : cattle manure and sawdust
53

55. Conclusion
 Vermicompost, vermicast and vermiwash are prepared by
earthworms
 Earthworms possess diverse microflora viz. bacteria, fungi and
actionomycetes in their gut
 Earthworms and microorganisms show symbiotic relationship
 The microbes carry out various enzyme activity and secrete
organic acids which digest agrowastes to simpler compounds that
can be easily taken up by earthworms to produce good quality
vermicompsot, vermicast and vermiwash
 Inoculation with such microbes enhance earthworm activity so as
to get higher biomass of earthworms resulting in improved quality
of product
54

56. Future thrust
 Studies are required to isolate and identify various microflora
present in earthworm gut
 Inoculation with microflora to get better quality vermicopmsot
and its products
55

57. THANK YOU . . .