Aquaculture Nutrition

2009 15; 513–516 doi: 10.1111/j.1365-2095.2008.00617.x

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1 2 3
1
Department of Zoology, Krishnagar Government College, Nadia, West Bengal, India; 2 Waste Management Cell, West Bengal
Pollution Control Board, Kolkata, West Bengal, India; 3 Department of Zoology, Krishnagar Government College, Nadia,
West Bengal, India

tendency to bioaccumulate toxic organic residues like pesti-

Three hundred adult healthy earthworms (Eudrillus euginae)
ranging length and weight from 14–29 cm (mean
21.5 ± 6.8 cm) to 1.6–3.0 g (mean 2.55 ± 1.1 g), respec-
tively, were cultured in three different substrate for 70 days.
The substrates were soil alone, (control T-1), cow (Bos
taurus) dung and water hyacinth (Eichhornia crassipes) (1 :
1 T-2), and partially dried neem (Azadirachta indica) leaves
with kitchen waste (1 : 1 T-3). The earthworms were
grown in these substrates in a wooden chamber
(0.8 m · 0.5 m · 0.2 m) and these were subjected to moni-
toring for various growth patterns (length, biomass, number)
at every fortnight (five observations) interval. The observa-
tions showed that T-2 was the best among the three treat-
ments used. There was significant difference (P < 0.05)
between the mean values of growth efficiency in food con-
version among the treatments. However, there was no sig-
nificant difference (P > 0.05) between the mean values of
survival rate of earthworm among different treatments.
cides, herbicides, and antibiotics and heavy metals like cad-
mium, nickel, lead and zinc into their tissue (Ramu 2001).
These toxicants can biomagnify their effects through use of
vermicompost. To avoid the bioaccumulation, hazard proper
substrate had to be used in vermiculture. There is an urgent
need of culture for earthworm, its utilization as live fish food,
bait, and fishmeal supplements (Akiyama et al. 1984;
Bankole et al. 1985; Omorinkoba et al. 1985; Stafford &
Tacon 1985; Tacon 1987; Chakrabarty 2008).
Eudrillus euginae is mostly used by the vermiculturist for
vermiculture. Several species of earthworms are being used
in vermiculture and sometimes two to three different spe-
cies of earthworms are simultaneously used for better
result. However, the present study deals with Eudrillus
euginae for vermiculture in variable substrates. The aim of
the study is to identify the suitable substrate for raising
earthworms.

KEY WORDS: earthworm, Eudrillus euginae, growth equation,

substrate, vermiculture, water hyacinth
Three rearing substrata namely soil, cow dung and water
Received 7 April 2008, accepted 25 June 2008
hyacinth, and partially dried neem leaves with kitchen waste
Correspondence: D. Chakrabarty, Department of Zoology, Krishnagar coded T-1, T-2, and T-3, respectively, were used for the
Government College, Krishnagar – 741101, Nadia, West Bengal, India.
E-mail: debajyoti_chakrabarty@yahoo.co.in
experiment.
Following was the composition details: three
replicates were used for each treatment in wooden
boxes (0.8 m · 0.5 m · 0.2 m) for culture of worms.
T.1 (control) Moist loamy-sandy soil with natural
Vermiculture, a century-old practice is now being revived
organic was used as soil substrate. This is most conven-
worldwide for waste management, sustainable organic agri-
tional substrate used for earthworm culture (Dynes 2003).
culture, and aquaculture (Springett & Syers 1979; Ismail
The sample was collected from nearby agricultural field
1997a; Sinha et al. 2002; Chakrabarty et al. 2007; Gupta
and properly grinded before application into wooden
et al. 2007; Chakrabarty 2008). But earthworms have high
boxes.
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© 2008 The Authors

Journal compilation © 2008 Blackwell Publishing Ltd
 T.2 Partially dried water hyacinth (30%) was mixed with cover)
fresh cow dung (70%) and allowed to decompose for 2 weeks following the method described by Dynes
before application. (2003). The total
T.3 The substrate was composed of neem leaves
(Azadirachta indica) and partially decomposes in kitchen
waste. The kitchen waste was mixed with neem leaves in
ratio of 1 : 1 on an outdoor earthen vat. Withered neem leaves
were settled and sun-dried. The neem leaves were soaked in
water for 5 days to retain the moisture for degradation into
compost. A change in the color of the leaves from pale brown
to dark after 7–9 days showed complete decomposition
(Jameson & Venkataramanujam 2002).

Three hundred adult earthworms ranging length and

weight from 1.6–3.0 g (mean 2.55 ± 1.1 g) to 14–29
cm (mean = 21.5 ± 6.8 cm), respectively, were
collected from the Akshay Krishi Vikash vermiculture
farm, Karimpur, Nadia. Rearing of earthworm was carried
out in nine woo- den boxes (0.8 m · 0.5 m · 0.2 m).
Three boxes were used for each substrate. The boxes were
screened with 3 mm wire screen net to protect the
earthworm from predators and other animals. The bottom
of the boxes were lined with polythene sheet. Then, the
earthworms were introduced at the rate of 33 worms per
box and covered with the substrate to a height of about 10
cm to form the second windrow (Dynes 2003). Water
spraying was performed with a watering can twice a day
during a sunny day and once in a cloudy day.

No extra feeding was given in this trial. Fresh substrate in

equal amount was supplied in each week after sampling.

The worms were sampled every fortnight. Wetting of the

substrate was stopped a day before the sampling day to make
the collection of the worms easier. The collection of worms
was performed by spreading neem leaves soaked in water
(for about 3 days) on top of the substrates. This served as
attractant and the earthworms came up at the top layer of the
substrate, from where they were collected by handpicking
using a forceps (Sogbesan & Ugwumba 2006).
Classification into different stages was performed using
age (counting culture days), weight (mg), and the presence
(or obviousness) and size of the clitellum (cocoon
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© 2008 The Authors

Journal compilation © 2008 Blackwell Publishing Ltd Aquaculture Nutrition 15; 513–516
population of worms from each substrate was counted and the total
weight was measured using a balance from the substrates. Worms
and clitellum's sizes were measured using a ruler.
The calculations were as follows:
Weight gainðWGÞ ¼ Final weight of worms produced
- initial weight of worms stocked

Relative Growth RateðRGRÞ

¼ ðWeight gain=initial body weightÞx 100

Specific Growth RateðSGRÞ ¼ ½ðlog wf - log wi Þ=t] x 100

where, log wf and log wi are logarithm of final weight and initial weight,
respectively, and t, the experimental period in days.
Reproductive performance (as Net Reproductive Rate or Nrr): This
was determined following Dynes (2003) and was used to compute
prediction equation
Nrr ¼ Total number of earthworms harvested=
½total number of earthworm stocked
x experimental periodðweeksÞ] x 100

All data recorded were computed and analyzed for significant difference
at P < 0.05 using single ANOVA. Regression anal- ysis of the data was also
computed. Graph pad in stat package was used on windows 2000 for all
the statistical analysis.

There was a gradual and continuous increase in the every fortnight

growth of the cultured earthworm from the three tested substrates
throughout the experimental period. In this present investigation, we
found a positive correlation between growth and culture period (r ‡
0.93) in each treat- ment. Four distinct developmental stages were
presented in all the three culture substrates. The developmental stages
were identified in the culture namely, baby, prejuvenile, juvenile, and
adult as shown in Table 1. Characteristic fea- tures and size of the
clitellum varied in the different stages of the earthworm. The baby
earthworm was devoid of clitellum. At the prejuvenile stage, the
appearance of the clitellum be- gan, although not well-ringed but had
faint reddish color with mean size of 1.5 ± 0.01 mm. At the juvenile
stage, the clitellum was swollen, well-ringed, and reddish color while
the adult clitellum size was 3.5 ± 0.08 mm and was swollen,
 Table 1 Developmental stages formation of clitellum of earthworm upto 70 days (t)

Number Length Clitellum's Description

Developmental stages Age (days) (%) (cm) Weight (mg) size/SD (mm) of clitellum

Baby 1–3 977 (44) 0.5–3.5 2.0–10.0 0 None

Pre-juvenile 4–6 643 (31) 3.6–3.9 11.0–90.0 1.5 ± 0.01 Not well-ringed, and
faint reddish color
Juvenile 7–15 317 (14) 4.0–10.0 91.0–1000.0 2.0 ± 0.05 Swollen, well-ringed,
and reddish color
Adult 16 Upward 215 (11) 14.0–29.0 1100.0–3000.0 3.5–0.08 Swollen enlarge and
brownish color
Initial number of ) 33 ) ) ) )
adult stocked per box

enlarged with brownish color. The highest average popula-
tion percentage (44.0%) was recorded as baby worm, while
the lowest population percentage (11.0%) was recorded as
adult in various substrates.
Highest weekly weight of 399.80 g earthworm kg)1
of substrate was recorded in T-2 followed by 366.4 g
earth- worm kg)1 substrate in T-3, while the lowest
weight of
305.2 g earthworm kg)1 of substrate was recorded in T-1.
However, the total quantities produced from the T-2 were
not significantly different (P > 0.05) from those produced
from T-3. The prediction equations for such production
are shown in Table 1.
Highest total weight gain of 308.2 g kg)1 of substrate was
recorded from earthworm cultured in T-2, while the
lowest value of 213.6 g kg)1 was recorded in T-1. There
was sig- nificant difference (P < 0.05) among treatments
with respect to the weight gain of earthworm culture in
T-1 and T-3 substrate (Table 2). The least relative growth
rate of 233.18% was recorded in earthworm culture in
soil substrate (T-1), while the highest value of 336.46%
was recorded in
earthworm culture in cellulose substrate (T-2). The highest
specific growth rate, 0.91% was recorded in T-2, while the
lowest 0.74% was recorded in T-1. The survival rate varied
between 98 and 99% for earthworm cultured in the three
substrates. There was no significant difference (P > 0.05)
between the mean values of survival rate of the earthworm
cultured using three different substrates.
Highest biomass production of 30.82 g earthworm week)1
was recorded in T-2, while the lowest value; 21.36 g earth-
worm week)1 was recorded in T-1 (Table 2). There was
significant difference (P < 0.05) of the biomass production
between T-2 and T-1 but no significant difference (P >
0.05) was observed between T-2 and T-3 substrate.
All the three culture substrates showed normal growth and
good survival of earthworm. Hence, these could be used as
commercial substrates for vermiculture. T-2 is the best
medium for earthworm culture than others. The significant

Table 2 Growth performance, nutrient

utilization productivity indices, and T.3 (Partially dry
prediction equation of earthworm using T-1 Soil T.2 (Cow dung neem leaves with
Parameters substrate and water kitchen waste)
(control) hyacinth)
different substrate in 70 days (t)
Initial total weight 91.60 91.60 91.60
of worms (g)
Final total weight 305.20* 399.80** 366.40*
of worms (g)
Total weight gain 213.60* 308.20** 274.80*
of worms (g)
Relative growth 233.18* 336.46** 296.44*
rate (%)
Specific growth rate 0.74* 0.91** 0.85**
Prediction equation W = 67.07 + 19.06t W = 63.13 + 14.42t W = 69.58 + 24.31t
R2 0.86 0.96 0.90
r 0.93 0.98 0.95
Average biomass 21.36* 30.82** 24.78*

W, weight; t, culture time in days.

Level shown as * P > 0, ** P < 0.05.
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© 2008 The Authors

Journal compilation © 2008 Blackwell Publishing Ltd Aquaculture Nutrition 15; 513–516
increase in the biomass production of earthworm compared
with other substrates shows favorable cocoon development
and hatching. A similar finding was reported by Aston
(1984). The best biomass production recorded in T-2 is in
agreement with the report of Omoyinmi et al. (2005), who
used sawdust and sand culture media for raising earthworms.
Another possible explanation for best growth and produc-
tion in T-2 is the production of bacteria involved in the
breaking down of cellulose and possible biochemical by-
products activated in presence of cow dung that are nat-
urally more numerous in T-2, when compared with other
substrates (Aston 1984). These bacteria are the protein
source for worms and needed in decomposition of organic
matter. Suzuki & Kurihara (1981) demonstrated that aquatic
oligochaete, Aelosoma hemprichi could feed exclusively on
bacteria, showing remarkable rapid population growth on
this diet. Williams et al. (2004) reported that the fecal
material produced by earthworms provides a ready supply of
labile organic substrates to the bacteria within soil and
compost, thus promoting microbial activity and their pro-
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1998; Ping & Boland 2004) that worms secrete enzymes,
including proteases, lipases, amylases, cellulases, and chitn-
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