chapter 5: screening of earthworm species for...

Chapter 5:

Screening of earthworm species for vermicomposting

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5.1 Introduction

The practice of vermiculture is at least a century old but it is now being received

worldwide with diverse ecological objectives such as waste management, soil

detoxification, regeneration and sustainable agriculture (Chauhan and Joshi, 2010). The

growth of industries and ever increasing human population has led to an increased

accumulation of waste materials (Joshi and Chauhan, 2006). The use of earthworms as a

waste treatment technique is gaining popularity and is commonly termed as

vermicomposting. Thus vermicomposting is an eco-biotechnological process that

transforms energy rich and complex organic substance into stabilized humus like product

‘vermicompost’ with the aid of earthworms (Garg et al., 2006a; Suthar and Singh, 2008).

Vermicomposting results in bioconversion of the waste into two useful products: the

earthworm biomass and the vermicompost. The former product can further be processed

into proteins (earthworm meal) or high-grade horticultural compost and the latter product

(vermicompost) is also considered as an excellent product since it is homogeneous, with

greatly increased surface areas and microsites for microbial decomposition and that

which tends to adsorb and retain nutrients over a longer period, without adversely

impacting the environment (Edwards et al., 2011). According to Sinha et al., (2010)

vermicomposting is “economically viable” (affordable by all nations), “environmentally

sustainable” (friendly to the environment-flora, fauna, soil, air and water, with no adverse

effect on them) and “socially acceptable” (beneficial to the society with no adverse effect

on human health) technology.

The actions of earthworms in the vermicomposting include substrate aeration,

mixing, grinding, fragmentation, enzymatic digestion and microbial digestion in

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intestines of earthworms (Sharma et al., 2005; Sinha, et al., 2010). Under the present

condition of environmental degradation ‘vermicomposting’ technology offers recovery of

valuable resources like ‘manure’ from such biodegradable waste. Recycling of wastes

through vermitechnology reduces the problem of dumping of huge quantities of wastes

and vermicompost has higher economic value compared with compost derived from

traditional methods (Chauhan et al., 2010).

Earthworms are voracious feeders on organic waste, converting a portion of the

organic material into worm biomass and respiration products and expel the remaining as

partially stabilized product (Benitez et al., 1999). Studies have also shown that

vermicomposting of organic waste accelerates organic matter stabilization (Neuhauser et

al., 1998) and gives chelating and phyto-hormonal elements (Tomati et al., 1995) which

have a high content of microbial matter besides stabilized humic substances. During

vermicomposting process the important plant nutrients such as nitrogen, potassium,

phosphorus and calcium present in feed material are converted into forms that are much

more soluble and available to the plants than those in the parent substrate (Ndegwa and

Thompson, 2001). Garg et al., (2006b) also reported increase in nitrogen, phosphorous

and potassium contents during vermicomposting. Since the intestine of earthworms

harbor wide range of microorganisms, enzymes and hormones, these half-digested

substrate (parent substrate) decomposes rapidly and is transformed into a form of

‘vermicompost’ within a short time (Lavelle, 1988).

5.2 Review of literature on screening of earthworms for vermicomposting

Earthworms of different species and ecological categories differ greatly in their ability to

digest various organic residues (Lattaud et al., 1998). Commonly adopted worms in

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vermiculture are Bimastos parvus, Dendrobaena rubida, D. veneta, Eisenia fetida, E.

hortensis and Eudrilus andrei, E. eugeniae, Amynthas diffringens, A. morrisi, Lampito

mauritii, Metaphire anomala, M. birmanica, Perionyx excavatus, P. sansibaricus,

Megascolex megascolex, Pontoscolex corethrurus, Octochaetona serrata, O. surensis,

Pheritima elongata, P. posthuma (Munnoli et al., 2010). But relatively few have been

used on a widespread scale and/or researched adequately.

The reproductive biology of P. excavatus, L. mauritii, P. elongata, Pontoscolex

corethrurus, E. gammiei, D. modiglianii and D. nepalensis (Dash and Senapati, 1980); D.

rubida and Lumbricus rubella (Elvira et al., 1996), E. andrei (Dominguez and Edwards,

1997), E. andrei and D. veneta (Fayolle et al., 1997), P. excavatus (Chaudhuri et al.,

2000); P. excavatus, Lampito mauritii, Polypheretima elongata, P. corethrurus,

Eutyphoeus gammiei, Dichogaster modiglianii and Drawida nepalensis (Bhattacharjee

and Chaudhuri, 2002); E. fetida and L. mauritii (Tripathi and Bhardwaj, 2004);

Metaphire posthuma (Bisht et al., 2007); P. ceylanensis (Karmegam and Daniel, 2009a);

Pontoscolex corethrurus, D. assamensis, D. papillifer papillifer, Eutyphoeus

comillahnus, Metaphire houlleti, Dichogaster affinis, Octochaetona beatrix, Lennogaster

chittagongensis (Chaudhuri and Bhattacharjee, 2011) has been studied by evaluating their

suitability for vermiculture.

Potential of some epigeic earthworms- Lumbricus terrestris, E. fetida, E. andrei,

Eudrilus eugeniae and P. excavatus to recycle organic waste materials into value-added

products is well documented (Kale et al., 1982; Elvira et al., 1998; Atiyeh et al., 2000;

Dominguez et al., 2001; Garg and Kaushik, 2005; Gajalakshmi et al., 2005; Tajbakhsh et

al., 2008; Navarro et al., 2009; Suthar, 2006, 2007 and 2009; Najar and Khan, 2010). Use

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of other species of earthworms (L. mauritii, Es eugeniae, Octochaetona serrata and

Perionyx excavatus, P. sansibaricus and Megascolex mauritii) are those of

Sathianarayanan and Khan (2007); Suthar and Singh (2008); Muthukumaravel et al.,

(2008); Karmegam and Daniel (2009b); Adi and Noor (2009); Bharadwaj (2010). Recent

reports by Paul et al., (2011) and Prakash et al., (2008) confirm that P. ceylanensis is a

potential vermicomposting species and the vermicompost produced by using this worm

had very good effect on plant growth and yield.

Among the epigeics, e.g. E. fetida, P. excavatus and Eudrilus eugeniae have

appeared as key candidates for organic waste recycling practices (Gajalakshmi et al.,

2002; Loh et al., 2005; Garg and Kaushik, 2005). However E. fetida is most commonly

used earthworm species for breaking down organic wastes because of its rapid growth

rate, reproductive potential, temperature tolerance range and its occurrence in organic

wastes with a wide range of moisture content (Edwards, 2004) and has been used for

recycling a wide variety of wastes (Kaviraj and Sharma 2003; Contreras-Ramos et al.,

2005; Garg et al., 2006b; Walkowiak, 2007; Tajbakhsh et al., 2008; Suthar 2009; Yadav

and Garg, 2011; Vig et al., 2011).

The presence of digestive enzymes like amylase, cellulase, protease, lipase,

chitinase have also been reported from the alimentary canal of earthworms (Munnoli et

al., 2010). Zhang et al., (1993) reported cellulase and mannase activities to be mainly due

to microorganisms. Mishra (1980) reported protease, amylase, cellulose, invertase and

urease in four species of indian earthworms, viz., Octochaetona surensis, L. mauritii, D.

calebi and Dichogaster balaui. Prabha et al., (2007) reported higher activity of amylase,

cellobiase, endoglucanase, acid phosphatase and nitrate reductase in the gut of E.

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eugeniae and E. fetida.

The decrease in C/N ratio, increase in concentration of N, P, K, phosphatase activity

in vermicast have been taken as criteria for judging the efficiency of earthworms in the

vermicomposting process (Garg et al., 2006a). In addition survival rate, biomass

production and reproduction of earthworms are the other indicator to evaluate the

vermicomposting process (Suthar, 2006).

5.3 Results

The data related to the experiments carried out to find the efficient/potential species for

the vermicomposting of macrophytes among the localy available earthworm species in

terms of their reproductive performance and the physicochemical characteristics of the

recycled product (vermicompost) are given below. The methodology adopted is presented

in chapter 2: Materials and methods, under subheads as earthworm cultures,

vermireactors and vermicasts.

Results of all the analyzed parameters of vermicasts obtained after recycling of

macrophytes by E. fetida, A. c. trapezoides and A. r. rosea during different time periods

(fortnights) are presented in Fig. 5.1.

pH ranged from 6.9 ± 0.06 to 7.7 ± 0.06, 6.9 ± 0.06 to 7.5 ± 0.05 and 6.7 ± 0.05 to

7.4 ± 0.06 in vermicasts of E. fetida, A. c. trapezoides and A. r. rosea respectively during

different fortnights (Fig. 5.1a).

Electrical conductivity indicated a value from 0.50 ± 0.02 to 0.71 ± 0.02 mS/cm and

0.45 ± 0.02 to 0.60 ± 0.02 mS/cm in E. fetida and A. c. trapezoides respectively as

compared to 0.40 ± 0.02 to 0.55 ± 0.02 mS/cm in vermicasts of A. r. rosea during

different fortnights (Fig. 5.1b).

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Concentration of potassium ranged from 29.33 ± 0.9 to 37.33 ± 0.9 mg/g, 27.33 ±

1.45 to 35 ± 0.6 mg/g and 19 ± 0.6 to 29.66 ± 1.2 mg/g in vermicasts of E. fetida, A. c.

trapezoides and A. r. rosea respectively during different fortnights (Fig. 5.1c).

Available phosphorous has higher values ranging from 456.43 ± 10.32 to 600 ±

7.93 µg/g in E. fetida as compared to 400 ± 10.50 to 523.66 ± 9.40 µg/g in A. c.

trapezoides and 324 ± 4.93 to 401.33 ± 10.17 µg/g in A. r. rosea vermicasts during

different fortnights (Fig. 5.1d).

Organic carbon decreased from 441.13 ± 14.10 to 156.23 ± 8.51 g/kg, 480 ± 11.54

to 246.66 ± 14.52 g/kg and 576.66 ± 14.52 to 280 ± 17.32 in vermicasts of E. fetida, A. c.

trapezoides and A. r. rosea respectively during different fortnights (Fig. 5.1e).

There was an increase in organic nitrogen from 4.97 ± 0.11 to 8.06 ± 0.32 g/kg;

5.34 ± 0.17 to 7.28 ± 0.24 g/kg and 5.53 ± 0.18 to 7 ± 0.17 g/kg in vermicasts of E.

fetida, A. c. trapezoides and A. r. rosea respectively during different fortnights (Fig. 5.1f).

Stability of C:N ratio was observed which ranged from 88.75 ± 2.44 to 19.38 ±

0.14, 89.88 ± 3.14 to 33.88 ± 2.63 and 104.27 ± 5.58 to 40 ± 1.37 in vermicasts of E.

fetida, A. c. trapezoides and A. r. rosea respectively during different fortnights (Fig.

5.1g).

ANOVA indicated significant variation among the vermicasts of the species and

during different time periods in pH (16.63; 76, P < 0.05), EC (86.04; 76.20, P < 0.05), K

(144.59; 70.80, P < 0.05), P (127.42; 33.95, P < 0.05), OC (63.31; 165.53, P < 0.05) and

C:N (238.36; 32.45, P < 0.05) ratio. However in case of ON significant variation (20.19,

P < 0.05) was observed only during different fortnights.

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Figure 5.1 3D graph showing characteristics of the vermicompost prepared from macrophytes during different fortnights by E. fetida, A. c. trapezoides and A. r. rosea.

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Figure 5.1 3D graph showing characteristics of the vermicompost prepared from macrophytes during different fortnights by E. fetida, A. c. trapezoides and A. r. rosea (continued).

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Vermicomposting experiment was also considered in terms of earthworm biomass

besides number of cocoons produced. Vermicomposting results in the conversion of a

part of the organic waste into worm biomass and respiratory products and other part of

the ingested is excreted as partially stabilized product i.e., vermicast (Benitez et al.,

1999).

Mean body weight was 0.64 ± 0.02 g in E. fetida, 0.53 ± 0.02 g in A. c. trapezoides

and 0.15 ± 0.01 g in A. r. rosea (Fig. 5.2a). Mean growth rate was 3.11 ± 0.28

mg/worm/day) in E. fetida, 0.38 ± 0.06 mg/worm/day in A. c. trapezoides and 0.15 ± 0.03

mg/day A. r. rosea (Fig. 5.2b). The percentage of relative growth rate was 14.74 ± 1.39

% in E. fetida; 1.93 ± 0.29 % in A. c. trapezoides and 1.19 ± 0.30 % in A. r. rosea (Fig.

5.2c).

Relative increase in earthworm number was 285 ± 7.63% in E. fetida, 95 ± 7.63%

in A. c. trapezoides and 45 ± 3% in A. r. rosea (Fig. 5.3a) which was significant (F =

6.58, P < 0.05) among the species, though there was no significant variation in number

during different fortnights. Increase in biomass was observed in E. fetida (69.16 ± 2.06%)

and in A. c. trapezoides (11.95 ± 1.12%), but a decrease was observed in A. r. rosea (8.82

± 2.53%) (Fig. 5.3b). Variation in biomass was significant (F = 21.03, P < 0.05) among

the species, with no significant variation (F = 2.97, P < 0.05) during different fortnights.

Cocoon production started after 15 days in both E. fetida and A. c. trapezoides, but

after 45 days in case of A. r. rosea, as the weed affect the reproductive potential of this

species and varied the cocoon production varies from 93 ± 6.06 to 174.7 ± 7.9 in E.

fetida, 11.33 ± 1.76 to 54 ± 2.08 in A. c. trapezoides and 1.67 ± 3.3 to 15 ± 1.73 in A. r.

rosea (Fig. 5.3c). Appearance of juveniles started after 30 days in E. fetida, after 45 days

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in A. c. trapezoides and after 60 days in A. r. rosea (Fig. 5.3d). Among these three species

after 60 days of experiment, the recycling potential (macrophytes) was 100% in the

epigeic E. fetida and it was 53.66 ± 0.88% and 33.66 ± 1% in the endogeic A. c.

trapezoides and A. r. rosea (Fig. 5.4) respectively.

Figure 5.2 Mean weight (a), growth rate (b) and relative growth rate (c) of earthworm species.

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

E. fetida A. c. trapezoides A. r. rosea

Mea

n w

eigh

t (g)

(a)

0

1

2

3

4


Gro

wth

rat

e m

g/w

orm

/day

(b)

0

5

10

15

20


Rel

ativ

e gr

owth

rat

e (%

)

(c)

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Figure 5.3 Error bars showing reproduction performances of E. fetida (EF), A. c. trapezoides (ACT) and A. r. rosea (ARR) on macrophytes during different fortnights.

333333333333N =

(a)

EF60

EF45

EF30

EF15

ARR60

ARR45

ARR30

ARR15

ACT60

ACT45

ACT30

ACT15

Incr

ease

in n

um

ber (%

)

400

300

200

100

0

-100

333333333333N =

(b)

EF60

EF45

EF30

EF15

ARR60

ARR45

ARR30

ARR15

ACT60

ACT45

ACT30

ACT15

Incr

ease

in b

iom

ass

(%

)

100

80

60

40

20

0

-20

-40

333333333333N =

(c)

EF60

EF45

EF30

EF15

ARR60

ARR45

ARR30

ARR15

ACT60

ACT45

ACT30

ACT15

Coco

ons

300

200

100

0

-100

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Figure 5.3 Error bars showing reproduction performances of E. fetida (EF), A. c. trapezoides (ACT) and A. r. rosea (ARR) on macrophytes during different fortnights (continued).

Figure 5.4 Error bar showing macrophytes recycling potential by E. fetida (EF), A. c. trapezoides (ACT) and A. r. rosea (ARR) during different fortnights.

5.4 Discussion

5.4.1 Characteristics of the vermicompost of E. fetida, A. c. trapezoides and A. r.

rosea

The marginal increase in pH with time interval from 6.9 ± 0.06 to 7.7 ± 0.06 in E. fetida

6.9 ± 0.06 to 7.5 ± 0.05 in A. C. trapezoids, 6.7 ± 0.05 to 7.4 ± 0.06 in A. r. rosea 6.7 ±

333333333333N =

(d)

EF60

EF45

EF30

EF15

ARR60

ARR45

ARR30

ARR15

ACT60

ACT45

ACT30

ACT15

Juve

nile

s

70

60

50

40

30

20

10

0

-10

333333333333N =

EF60

EF45

EF30

EF15

ARR60

ARR45

ARR30

ARR15

ACT60

ACT45

ACT30

ACT15

Recy

cling o

f macrophytes (%

) 120

100

80

60

40

20

0

-20

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0.05 was observed and the overall increase in pH could be due the decomposition of

ammonia, which forms a larger proportion of nitrogenous matter excreted by earthworms

(Muthukumaravel et al., (2008).

Electrical conductivity indicated increasing trend from first fortnight to fourth

fortnight but maximium increase was in E. fetida-mediated treatment (0.50 ± 0.02 to 0.71

± 0.02 mS/cm) and minimum in A. r. rosea treatment (0.40 ± 0.02 to 0.55 ± 0.02 mS/cm).

The results corroborate the study of Kaviraj and Sharma (2003) where an increase in EC

of vermicast during time interval was reported and is attributed to the loss of organic

matter and release of different mineral salts in available form such as phosphate,

ammonia and potassium (Najar and Khan, 2010).

Among the treatments maximium decrease from 441.13 ± 14.10 to 156.23 ± 8.51

g/kg in organic carbon observed in E. fetida-mediated treatments is consistent with the

studies of Garg and Kaushik (2005) and Suthar (2007). Goyal et al., (2005) reported that

during vermicomposting a large fraction of organic matter in substrates is lost as carbon-

dioxide.

Though increase in organic nitrogen was observed over different time intervals,

with maximium in E. fetida (4.97 ± 0.11 to 8.06 ± 0.32 g/kg) and A. c. trapezoids (5.53 ±

0.18 to 7 ± 0.17 g/kg) treatments, the observed differences among the 3 species could be

attributed directly to the feeding preferences of the epigeic and endogeic earthworm

species and indirectly to mutualistic relationship between ingested microorganisms and

intestinal mucus which might be species-specific (Suthar and Singh, 2008).

The concentration of potassium in the vermicast of the epigeic E. fetida (37.33 ± 0.9

mg/g) is higher than the endogeic A. c. trapezoides (35 ± 0.6 mg/g) and A. r. rosea, which

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is attributed to the production of carbonic, nitric and sulphuric acids by microorganisms

present in the gut of earthworms (Kaviraj and Sharma, 2003).

Phosphorus content was higher after 60 days in all three treatments, being highest in

E. fetida treatment (600 ± 7.93 µg/g) followed by A. c. trapezoides (523.66 ± 9.40 µg/g)

but was least in A. r. rosea (401.33 ± 10.17 µg/g). Garg et al., (2006b) reported increase

in concentration of phosphorous during vermicomposting. The enhanced phosphorous

level in vermicompost is probably through mineralization and mobilization of phosphorus

by bacterial and faecal phosphatase activity of earthworms (Jeyanthi et al., 2010).

Stability of C:N ratio during vermicomposting is attributed to the loss of carbon as

carbon dioxide in the process of respiration and production of mucus and nitrogenous

excrements that enhance the level of nitrogen (Hayawin et al., 2010). Castillo et al.,

(2010) reported that a decline in C:N ratio to less than 20 indicates an advanced degree of

organic matter stabilization and it reflects a satisfactory degree of maturity of organic

wastes. The low C:N ratio (19.3 ± 0.14) from the epigeic E. fetida processed treatment

indicates that this species enhances the organic matter mineralization more efficiently

than the other two endogeics- A. c. trapezoides and A. r. rosea.

The observed difference in the nutrient contents of vermicasts among the species is

attributed to feeding habit which determines the nature and properties of vermicasts

produced (Haynes, 2003).

5.4.2 Reproductive performance of E. fetida, A. c. trapezoides and A. r. rosea

At the end of the experiment (60 days) increase in earthworm number was higher in

epigeic E. fetida followed by A. c. trapezoides but was least in the other endogeic A. r.

rosea. Consumption of weed with time provides nutrients that enhance earthworm

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reproductive capability leading to increase in number (including juveniles) by the end of

60 days. However it is evident that the variation in growth rates among the species

observed is related to species-specific feeding habitat. Saini et al., (2010) reported growth

and reproduction in E. fetida as rapid when compared to other species.

Higher production of cocoon was observed in E. fetida reactor as compared to that

of A. c. trapezoides and A. r. rosea during all fortnights. Chauhan et al., (2010) reported a

varied number of cocoon production during the vermicomposting by using E. fetida, E.

eugeniae and P. excavatus and is said to be species to substrate specific.

5.4.3 Recycling of macrophytes during different fortnights by E. fetida, A. c.

trapezoides and A. r. rosea

Different species of earthworms can show distinct preference for plant material (Curry

and Schmidt, 2007). Efficient recycling of macrophytes over the period of time is shown

by epigeic E. fetida (100%) than the other two endogeic species. The observed difference

in recycling of macrophytes between E. fetida, A. c. trapezoides and A. r. rosea could be

related to the feed preferences based on the ecology of individual earthworm species

(Suthar and Singh 2008; Indrajeet et al., 2010).

5.5 Conclusion

From the apparent discussion it could be concluded that the epigeic earthworm species E.

fetida has a higher potential in recycling of macrophytes than the two endogeic species-

A. c. trapezoides and A. r. rosea. Further the data manifests vermicast of E. fetida as rich

in plant nutrients relative to the other two species. Moreover, its growth was higher and

reproductive behaviour favorable on this substrate. The composting potential besides

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being a species-specific character, is also related to the feeding preferences of epigeic and

endogeic composting earthworms. Species which are capable of dwelling in high

percentage of organic material along with high adaptability to environmental changes,

with high fecundity rate, high rate of consumption, digestion, assimilation and growth

possess a better potential for vermicomposting process. Thus the study revealed that the

resourceful efficiency of E. fetida should be used to combat noxious macrophytes

invasion of the lakes into value-added materials, i,e vermicompost besides earthworm

biomass.

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chapter 5: screening of earthworm species for...

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