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Growth, reproductive biology and life cycle of the vermicomposting earthworm, Perionyx ceylanensis Mich. (Oligochaeta: Megascolecidae) ARTICLE in BIORESOURCE TECHNOLOGY · JUNE 2009 Impact Factor: 4.49 · DOI: 10.1016/j.biortech.2009.05.004 · Source: PubMed

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Bioresource Technology 100 (2009) 4790–4796

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Bioresource Technology journal homepage: www.elsevier.com/locate/biortech

Growth, reproductive biology and life cycle of the vermicomposting earthworm, Perionyx ceylanensis Mich. (Oligochaeta: Megascolecidae) Natchimuthu Karmegam a,*, Thilagavathy Daniel b a b

Department of Biotechnology, V.M.K.V. Engineering College, Vinayaka Missions University, Periya Seeragapadi, Salem 636 308, Tamil Nadu, India Department of Biology, Gandhigram Rural University, Gandhigram 624 302, Dindigul District, Tamil Nadu, India

a r t i c l e

i n f o

Article history: Received 18 February 2009 Received in revised form 4 May 2009 Accepted 5 May 2009 Available online 24 May 2009 Keywords: Cocoons Earthworm Hatchlings Parthenogenesis Vermicomposting

a b s t r a c t In the present study, an attempt has been made to study the growth, reproduction and life cycle of the earthworm, Perionyx ceylanensis Mich. in cowdung for the period of 340 days. Results showed that the overall mean growth rate was 1.79, 1.57 and 1.34 mg/worm/day respectively for the worms cultured singly, in batches of four and eight. Cocoon production rate was found between 0.85 and 0.94 cocoons/ worm/day and the hatching success between 74.67% and 82.67%. The majority of the cocoons (95.16– 96.77%) hatched only one hatchling. Worms raised singly also produced viable cocoons indicating that P. ceylanensis reproduce parthenogenetically. The life cycle of the worms cultured singly was ±57 days and it was ±50 days for the worms cultured in batches of four and eight. There is a vast scope to utilize P. ceylanensis for vermiculture practices due to short period of life cycle. Ó 2009 Elsevier Ltd. All rights reserved.

1. Introduction Reproduction in earthworms is not only peculiar because of hermaphroditism but also because there is a paucity of information in this regard. It is absolutely essential to study the growth and development of earthworms in order to promote vermiculture based biotechnology. This may help in understanding the organism, selecting prolific breeders, predicting the population characteristics (in activity or in nature) with reference to feeding habits, temperature, pH and moisture tolerance and reproduction rates in order to evaluate their vermicomposting potential (Julka et al., 2009). Some aspects of the reproductive biology of Indian earthworms have been studied for species such as Perionyx excavatus (Chaudhuri and Bhattacharjee, 2002; Suthar and Ram, 2008; Suthar, 2009), Lampito mauritii (Bhattacharjee and Chaudhuri, 2002; Tripathi and Bhardwaj, 2004) and Perionyx sansibaricus (Suthar 2007a,b, 2009). Apart from P. excavatus and P. sansibaricus, other Perionyx species with vermicomposting potential include P. bainii, P. ceylanensis and P. nainianus which are native worms distributed in India (Julka et al., 2009), but studies on their reproductive potential is insufficient. The recent reports by Karmegam et al. (2003), Prakash et al. (2008), Karmegam and Daniel (2008a,b), and John Paul et al. (2008) undoubtedly confirm that the earthworm species, P. ceylan-

* Corresponding author. Tel.: +91 427 2477218; fax: +91 427 2477919. E-mail addresses: [email protected], [email protected] (N. Karmegam). 0960-8524/$ - see front matter Ó 2009 Elsevier Ltd. All rights reserved. doi:10.1016/j.biortech.2009.05.004

ensis is a potential vermicomposting species and the vermicompost produced by using this worm had very good effect on plant growth and yield. While investigating the efficiency of L. mauritii and P. ceylanensis for vermicomposting different organic substrates, Karmegam and Daniel (2008b) suggested that both the earthworm species can be used for vermicomposting; however, duration of vermicomposting with P. ceylanensis is not as much of L. mauritii. The standardization of P. ceylanensis, a locally available species, for vermicomposting of different organic substrates is a new finding and the species could be useful for vermiconversion of organic substrates under local conditions. A study carried out by Prakash et al. (2008) showed that the vermicasts produced by the earthworm species, P. ceylanensis reared in Polyalthia longifolia leaf liter + cowdung (1:1) contained 14 different fungal species belonging to the genera, Aspergillus, Chaetomium, Cladosporium, Cunninghamella, Fusarium, Mucor, Penicillium and Rhizopus. Total nitrogen, phosphorus, potassium, calcium, copper, iron and zinc were higher in vermicasts than control (worm-unworked substrate) while organic carbon and C/N ratio were lower in vermicasts. Field trials conducted by Karmegam and Daniel (2008a) with Lablab purpureus (L.) Sweet. showed that growth and yield parameters were significantly higher in the plots which received vermicompost, chemical fertilizer and vermicompost + chemical fertilizer mixture than in the control plots (p < 0.05). The highest fruit yield (fresh weight) of 109 tonnes ha 1 was recorded in the treatment which received 2.5 tonnes of vermicompost [prepared with a weed, Rottboellia exaltata + cowdung, (1:1) using P. ceylanensis] + 1/2 dose of recommended NPK ha 1, while it was 61.9 tonnes ha 1 in control plots without vermicompost and/or chemical fertilizer.


N. Karmegam, T. Daniel / Bioresource Technology 100 (2009) 4790–4796

Several earthworm species have been identified as detritus feeders that can be reared in large numbers on organic waste material. Some of these epigeic species have been shown to be suitable candidates for vermiculture, where organic waste is transformed into useful compost, e.g. Eisenia fetida, Eudrilus eugeniae, P. excavatus, P. sansibaricus and P. ceylanensis (Dominguez et al., 2001; Tripathi and Bhardwaj, 2004; Meena and Renu, 2008; Karmegam and Daniel, 2008a; Suthar, 2006, 2007b,c; Suthar and Ram, 2008; Suthar and Singh 2008). The growth and reproductive studies with regard to P. ceylanensis is unavailable. Hence the present study is focused on the growth, reproductive biology and life cycle of the earthworm, P. ceylanensis.

2. Methods 2.1. Mass culture of P. ceylanensis Clitellate adult worms of P. ceylanensis were cultured in mass culture tanks containing cowdung medium in the Department of Biology, Gandhigram Rural Institute, Gandhigram, from the stock originally collected from manure pits in and around Gandhigram Rural Institute and from litter heaps in Sirumalai Hills.

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authentication. For the growth study, the hatchlings were gathered from replidishes in which cocoons had been placed to hatch. The hatchlings were raised singly and in batches of four and of eight respectively. Since the number of earthworms in growth medium was reported to influence growth and reproduction by Kaushal et al. (1995), worms were raised singly and in batches of four and eight in the present study also. The number of replicates for the hatchlings kept singly was 30, while for batches of four and of eight hatchlings, it was sixteen. The biomass and length of each hatchling were measured and then introduced to the experimental containers which were kept in a rearing chamber at 27 ± 1 °C with a relative humidity of 80–85%. After 15 days, fresh medium was added to every container in quantities related to the number of worms reared in the container. This was repeated every ten days for 100 days and, after that, for every five days. While adding the fresh medium some of the older substrates were removed. This was done to maintain the original volume of the medium in each container. The biomass and the length of P. ceylanensis were determined every seven days up to 70th day. Subsequent weighing and measuring were performed every 15 days up to 160 days. Afterwards, till the end of the experiments, the biomass and length of the worms were determined every 30 days. The growth rate of the worms was calculated as per the formula of Mazantseva (1982).

2.2. Earthworm breeding and collection of cocoons 2.5. Reproduction and life cycle of P. ceylanensis From the mass culture tanks, fifty (50) healthy adult worms of uniform size were sorted out and transferred to plastic troughs of 45 cm 30 cm 15 cm size filled 2/3 with cowdung holding 70–80% moisture content. A total of ten replicates were maintained. After ten days from the introduction of worms, the vermibeds were searched for cocoons with the help of magnifying lens. The cocoons collected were washed with tap water and kept in Petri dishes containing distilled water and incubated. Soon after the emergence of young worms i.e., hatchlings, they were used for further experiments.

A batch of six replicates was separately kept for hatchlings raised singly and in batches of four and of eight P. ceylanensis, for studying the life-cycle parameters. The worms raised singly and in batches were closely watched for the development of clitellum after 15 days of the introduction of the worms until all the worms developed clitellum. After the formation of clitellum the substrates were searched for cocoons for every five days and the number of cocoons produced was recorded for each experimental set-up till the termination of the study.

2.3. Preparation of growing medium (substrate)

2.6. Incubation of cocoons

For all the studies i.e., growth and reproductive studies, cowdung was used as the growing medium. The hatchlings and growing worms were maintained in cowdung as this was deemed a standard medium by Reinecke and Hallatt (1989) and Reinecke et al. (1992) and others. The cowdung (pH 7.3, organic carbon27.2%, total nitrogen-1.5%, total phosphorus-0.9% and total potassium-1.2%) used in the study was collected from the University dairy farm, shade dried and finely powdered. Thereafter the finely powdered material was sieved to a particle size of 1000–500 lm. Since this size was found to be favourable for raising of young earthworms of P. excavatus (Reinecke and Reinecke, 1994), the same size was used for the present study also. Distilled water was added to the powder to prepare a dampened medium with a moisture content of 70–80% and allowed to stabilize for 48 h in a rearing chamber where the relative humidity was 80%. After stabilization a few worms were placed in the dampened medium and observed for any mortality for 48 h. If no mortality occurred within 48 h, the medium was deemed fit for the experimental animals.

The cocoons recovered from the vermibeds produced by P. ceylanensis were kept in small containers with dampened cowdung. From the total cocoons recovered, a batch of 75 cocoons were randomly selected and weighed. The cocoons were individually kept in small plastic containers of 2.5 cm dia 2.5 cm height carrying one gram of medium and observed for the emergence of hatchlings. The number of hatchlings (young worms) that emerged from each cocoon and the total number of cocoons that hatched were recorded. The biomass of hatchlings was also recorded by digital analor balance. The percentage of hatching success was calculated as per standard methods. The biomass of the cocoons and the biomass of hatchlings were statistically correlated using computer software, Microcal Origin (ver. 3.1).

2.4. Growth studies

The growth rate of P. ceylanensis reared singly and in batches of four and of eight is given in Table 1. The average initial weight of the worm was 1.63 ± 0.13 mg. The growth rate was very low during the first few days of growth (1–14 days) and then the rate of growth increased (21–28 days). Again the rate of growth decreased and then increased. This trend was observed throughout the period of study. The rate of growth did not follow any fixed pattern. The highest growth rate recorded was 5.61, 5.33 and 4.93 mg/worm/

The containers used for the study measured 12 cm dia 9 cm height and were fitted with perforated lids. Initially 50 gm of the medium was kept in each container. As suggested by Hallatt et al. (1990), the work was started with many replications and, afterwards, restricted to minimum numbers (as given below for each experiment) in order to get accurate results and for statistical

3. Results and discussion 3.1. Growth of P. ceylanensis


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Table 1 Growth rate (mean ± S.D.) of P. ceylanensis cultured singly, in batches of four and of eight in cowdung medium (340 days). Period of growth (d)

Growth rate of worms (mg/worm/day) Cultured singly

Cultured in batches of four

Cultured in batches of eight

0–28 29–56 57–85 86–145 146–340

3.53 ± 0.32 4.02 ± 0.35 2.24 ± 0.19 2.27 ± 0.31 1.01 ± 0.12

3.12 ± 0.28 3.57 ± 0.30 2.03 ± 0.23 1.97 ± 0.15 0.88 ± 0.10

2.84 ± 0.21 3.11 ± 0.24 1.57 ± 0.16 2.02 ± 0.20 0.63 ± 0.08

700

Worm biomass (mg)

600

700

600

r=0.989

Worm biomass (mg)

500

r=0.966 r=0.987

400

300

200 Worms cultured singly (——) Worms cultured in batches of four (— — —) Worms cultured in batches of eight (- - -)

100

0 0

100

200

300

400

Age (days) Fig. 2. Growth curve (biomass) of P. ceylanensis cultured singly, in batches of four and of eight in cowdung medium (340 days) (error bars indicate ± S.D.).

10

8

Length of worms (cm)

day at the age of 21–28 days for the worms cultured singly, in batches of four and of eight, respectively. The mean growth rate of the worms cultured singly, in batches of four and of eight was 1.79, 1.57 and 1.34 mg/worm/day respectively for the entire period of study i.e. 340 days. The hierarchy of the growth rate of the worms was: worms cultured singly > batches of four > batches of eight. The initial mean biomass of the hatchlings subjected to 340 days growth study was 1.63 ± 0.13 mg/worm. The biomass increase was high up to 145 days and then it was less (Fig. 1). The worms cultured singly reached a mean biomass of 413.87 ± 18.64 mg/worm at the age of 130 days. The age at which the worms cultured in batches of four and of eight reached more than 400 mg/worm was 190–220 days for worms cultured in batches of four and 220 to 250 days for worms cultured in batches of eight. The total biomass gained during the 340 day study period was 608.21 ± 38.66, 534.13 ± 42.71 and 465.37 ± 41.80 mg/worm for the worms cultured singly, in batches of four and of eight, respectively. More than threefold biomass increase was observed during the initial seven days. It reached an average of 266.15, 201.07 and 191.08 folds on the 100th day and 467.85, 381.50 and 349.23 fold on the 340th day, for the worms cultured singly and in batches of four and of eight respectively. Fig. 2 shows the growth curve of worm biomass. A highly significant positive correlation at 0.1% was observed between the age and the biomass of the worms (p < 0.001). The results of the body length of P. ceylanensis cultured in cowdung for 340 days are given in Fig. 3. The initial length of the hatchling subjected to the growth study was 0.51 ± 0.05 cm. Till the 70th day the body length of the worms showed a tremendous increase and after that the increase in the length was very little. Towards

6

4

500

2 Worms cultured singly

400

Worms cultured in batches of four Worms cultured in batches of eight

0

300

0

50

100

150

200

250

300

350

400

Age (days) 200

Fig. 3. Body length of P. ceylanensis cultured singly, in batches of four and of eight in cowdung medium (340 days).

Worms cultured singly Worms cultured in batches of four

100

Worms cultured in batches of eight

0 0

50

100

150

200

250

300

350

400

Age (days) Fig. 1. Biomass of P. ceylanensis cultured singly, in batches of four and of eight in cowdung medium (340 days).

the end of the study period a slight decline in length was observed. In the initial period, i.e. 1–7 days, threefold increase was observed in the body length of all the worms, i.e. the worms cultured singly, in batches of four and of eight. The highest length was recorded at the age of 190 days, when the worms showed an increase of 15.80,



14.51 and 15.14 fold for the worms cultured singly, in batches of four and of eight respectively. On the 340th day the body length of the worms was 6.61, 6.50 and 6.12 cm for the worms cultured singly, in batches of four and of eight, respectively. Increase in the body length was marked during the initial days (1–75 days). Then the length was almost stable up to the 200th day and then it started to decrease (Fig. 3). The growth curve of body length indicated a positive correlation between the age and the worm length (Fig. 4). The length of the worms also showed a perfect correlation with the worm biomass, with a significant level of 0.1% (p < 0.001). The correlation co-efficient (r) of 0.786, 0.793 and 0.781 were observed between worm length and worm biomass for the worms raised singly, in batches of four and in batches of eight, respectively. The growth rate of P. ceylanensis cultured singly and in batches of four and of eight showed an average rate of 1.34–1.79 mg/ worm/day (Table 1). The body length of P. ceylanensis steadily increased up to the 200th day and then it declined (Figs. 3 and 4). The length of the worms showed a perfect correlation with the worm biomass (worms cultured singly: r = 0.786, y = 0.0081x + 3.807; worms culture in batches of four: r = 0.793, y = 0.0096x + 3.562; worms cultured in batches of eight: r = 0.781, y = 0.0105x + 3.527). Kaushal et al. (1995) observed in D. nepalensis grown singly in oak litter and in pine litter, a higher biomass than in the worms grown in batches. A growth rate of 10.6 and 5.5 mg/ adult/day was recorded for the worms grown singly and in batches respectively, in oak litter medium. Chaudhuri and Bhattacharjee (2002) reported that the rate of biomass increase of P. excavatus was maximum in the mixtures with straw and bamboo leaf litter and concluded that the cowdung, a natural food of P. excavatus, was marginally better than the mixture with kitchen waste with regard to the rate of biomass increase and reproduction. A 105 day study with different experimental diets on the biomass increase of P. excavatus showed that the overall rate of biomass production was maximum, 4.75 mg/adult/day in cowdung-straw medium where as the rate of biomass production was 2.86 mg/ adult/day. Suthar (2007c) reported a maximum growth rate by P. excavatus between the range of 3.69–2.35 mg/worm/day on different feed mixtures, crop residue mixed with cattle solids, farm yard 10 r=0.803

9 8

Length of worms (cm)

7 6 r=0.819 5

r=0.822

4 3 2 Worms cultured singly (——) Worms cultured in batches of four (— — —)

1

Worms cultured in batches of eight (- - -)

0 0

30

60

90

120

150

180

210

240

270

300

330

360

Age (days) Fig. 4. Growth curve (length) of P. ceylanensis cultured singly, in batches of four and of eight in cowdung medium (340 days) (error bars indicate ± S.D.).

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manure and municipal solid waste. Recent investigation by Suthar (2009) on the growth and fecundity of earthworms, P. excavatus and P. sansibaricus in cattle waste solids showed a consistent trend of rapid biomass increase up to 13 week. Then both the earthworm species showed a marked stabilization followed by gradual decline in growth rate at the end. Suthar (2009) also observed significantly higher individual live of weight of 767.7 mg for P. sansibaricus than P. excavatus (612.6 mg) after 13 weeks of study. In the present study, the growth rate of P. ceylanensis raised singly and in batches falls between 1.3 and 1.8 mg/worm/day where these values were lower than the growth rates recorded for P. excavatus and P. sansibaricus by other workers which ranged from 3.5 to 8.0 mg/worm/ day (Reinecke et al., 1992; Bhattacharjee and Chaudhuri, 2002; Suthar and Ram, 2008; Suthar, 2009). This may be due to the feed material, feeding preference of the worms and duration of the study. The growth rate of P. ceylanensis indicates that this species is a suitable candidate for mass culture. 3.2. Reproduction and life-cycle of P. ceylanensis The development of clitellum in P. ceylanensis started on the 18th day for the worms cultured singly and on the 15th day for the worms cultured in batches of four and of eight. Development of the clitellum continued up to the 35th day for the worms grown singly and up to the 31st day for the worms cultured in batches of four and of eight. The mean age of clitellum development was 25.57, 22.14 and 22.00 days for the worms cultured singly and in batches of four and of eight, respectively (Table 2). The rate of cocoon production of P. ceylanensis cultured singly and in batches of four and of eight is given in Fig. 5. The rate of cocoon production was high up to 70 days and thereafter until the end of the observation, i.e. 160 days, there was not much change in the rate of production of cocoons among the worms cultured singly and in batches of four and of eight (Fig. 5). The first appearance of the cocoon was found at the age of 23 days. The worms that were reared singly also produced cocoons. The rate of cocoon production was 0.28, 0.32 and 0.30 cocoon/worm/day during the age of 23–45 days for the worms cultured singly, in batches of four and of eight respectively. The highest rate of production of cocoon was observed during the age of 120–150 days and it was 1.18, 1.31 and 1.26 cocoons/worm/day for the worms cultured singly, in batches of four and of eight respectively. During 250–340 days, the cocoon production rate was 0.97, 1.08 and 1.04 cocoons/worm/day for the worms cultured singly, in batches of four and of eight respectively. The overall cocoon production rate was 0.85, 0.94 and 0.91 cocoons/worm/day for the worms cultured singly, in batches of four and of eight respectively. Incubation period of the cocoons of P. ceylanensis ranged from 12– 26 days. Out of the 75 cocoons incubated from each experimental setup, 56, 62 and 62 cocoons hatched from the cocoons produced by the worms cultured singly, in batches of four and of eight, respectively. A hatching success of 74.67, 82.67 and 82.67% was recorded for the cocoons produced by the worms cultured singly, in batches four and of eight, respectively (Table 3). The largest number of cocoons hatched during 17–21 days (Table 4).The majority of the cocoons, i.e. 95.16–96.77%, hatched only one hatchling (Table 5). The average biomass of cocoons was 6.48, 6.58 and 6.60 mg/cocoon produced by the worms cultured singly, in batches of four and of eight, respectively; the respective maximum individual cocoon biomass was 9.10, 9.50 and 10.10 mg. The average biomass of hatchlings that emerged from the cocoons produced by the worms cultured singly, in batches of four and of eight was 1.65, 1.70 and 1.65 mg, respectively. The highest individual biomass of the hatchling was 2.40, 2.85 and 2.76 mg for the hatchlings of worms raised singly, in batches of four and of eight, respectively. It was observed


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Table 2 Development of clitellum in P. ceylanensis cultured singly, in batches of four and of eight in cowdung medium. Treatment

Age (days)

Number of clitellate worms

% Clitellate worms

Worms cultured singly

18 20 22 25 28 31 35 25.57 ± 6.13

1 5 12 27 37 38 40

2.50 12.50 30.00 67.50 92.50 95.00 100.00

15 17 19 21 24 28 31 22.14 ± 5.84

3 9 28 35 38 39 40

7.50 22.50 70.00 87.50 95.00 97.50 100.00

15 17 19 21 24 28 30 22.00 ± 5.60

2 10 22 37 38 39 40

2.50 25.00 55.00 92.50 95.00 97.50 100.00

Mean ± S.D. Worms cultured in batches of four

Mean ± S.D. Worms cultured in batches of eight

Mean ± S.D.

1.4

No. of cocoons/worm/day

1.2

1.0

0.8

0.6

0.4 Worms cultured singly Worms cultured in batches of four Worms cultured in batches of eight

0.2

0.0 0

50

100

150

200

250

300

350

400

Age (days) Fig. 5. Rate of cocoon production (no/worm/day) of P. ceylanensis cultured singly, in batches of four and of eight in cowdung medium (340 days).

that the cocoon biomass was directly proportional to the biomass of hatchlings and the cocoon biomass showed significant positive correlation (p < 0.01) with the hatchling biomass (r = 0.798 for the worms cultured singly; r = 0.881 for the worms cultured in batches of four; r = 0.862 for the worms cultured in batches of eight). The duration of the life cycle of the worms cultured singly was ±57 days and it was ±50 days for the worms cultured in batches of four and of eight (Table 6). Development of clitellum is a very important feature in the process of reproduction in earthworms. The number of cocoons produced/adult/day, individual cocoon weight, and total number of cocoons produced/adult were higher for the worms grown singly than in batches in the present study. The number of days taken for clitellum development ranged from 15 to 35 days for P. ceylanensis (Table 2). The rate of cocoon production was 0.85 to 0.94 cocoon/worm/day (Fig. 5). A fecundity rate of 156 cocoons/adult/year under laboratory conditions was recorded for the epigeic earthworm, P. excavatus by Bhattacharjee and Chaudhuri (2002). The fecundity rate of P. ceylanensis cultured singly in cowdung was lower than that of the cocoon production rate recorded for the worms cultured in batches of four and eight. It is very clear that the fecundity of P. ceylanensis in the present study is higher than

Table 3 Hatching of the cocoons of P. ceylanensis cultured singly, in batches of four and of eight in cowdung medium. Parameters observed

Worms cultured Singly

In batches of four

In batches of eight

No. of cocoons incubated

75

75

75

No. of cocoons hatched with One hatchling Two hatchlings Total no. of cocoons hatched Hatching success (%)

54 2 56 74.67

59 3 62 82.67

60 2 62 82.67

Mean (±S.D.) no. of hatchlings Of total no. of cocoons incubated Of total no. of cocoons hatched Mean (±S.D.) biomass of cocoons (mg) Mean (±S.D.) biomass of hatchlings (mg)

0.67 ± 0.53 0.96 ± 0.34 6.48 ± 1.34 1.65 ± 0.34

0.76 ± 0.23 1.06 ± 0.23 6.58 ± 1.39 1.70 ± 0.44

0.75 ± 0.50 1.04 ± 0.19 1.70 ± 0.44 1.65 ± 0.45


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N. Karmegam, T. Daniel / Bioresource Technology 100 (2009) 4790–4796 Table 4 Hatching pattern of cocoons of P. ceylanensis cultured singly, in batches of four and of eight in cowdung medium during incubation. Incubation period (days)

Hatching of cocoons (no. in%) produced by

12 13 14 15 16 17 18 19 20 21 22 23 24 25 26

Worms cultured singly

Worms cultured in batches of four

Worms cultured in batches of eight

1.79 1.79 0 3.57 3.57 8.93 14.29 21.43 25.00 16.07 0 1.79 0 0 1.79

1.61 3.23 0 6.45 4.84 20.97 25.81 19.35 9.68 4.84 0 0 1.61 1.61 0

3.23 0 1.61 3.23 6.45 22.58 27.42 17.74 8.06 3.23 3.23 1.61 0 1.61 0

Table 5 Number of hatchlings (%) emerged from the cocoons of P. ceylanensis cultured singly, in batches of four and of eight in cowdung medium. Cocoons produced by

No. of hatchlings emerged (%)/cocoon

Worms cultured singly Worms cultured in batches of four Worms cultured in batches of eight

One

Two

96.43 95.16 96.77

3.57 4.84 3.23

that recorded for P. excavatus and P. sansibaricus by various studies with the exception being 7.70 cocoons/worm/week for P. excavatus in cattle manure (Reinecke et al., 1992) (Table 7). Worms at lower densities constantly increased their production and maintained higher rate. In vermicomposting system, it is optimized that the vermibed containing 4 kg of organic materials can hold about 90–120 P. ceylanensis for efficient vermiconversion (Karmegam and Daniel 2008b). In the present study also, in defined medium, i.e., cowdung, this trend was observed in cultures of P.

Table 6 Lifecycle of P. ceylanensis cultured singly, in batches of four and of eight in cowdung medium. Sl. No.

Life-cycle parameters

Worms cultured

1 2 3 4 5 6 7 8 9-

Appearance of clitellum (days) Commencement of cocoon production (days) Rate of cocoon production (cocoon/worm/day) Incubation period (days) Hatching success (%) Mean no. of hatchlings (per cocoon) No. of hatchlings produced from one cocoon Duration of life-cycle (days) Highest individual worm biomass (mean) (mg/worm)

Singly

In batches of four

In batches of eight

18–35 23–40 0.85 ± 0.21 19.00 ± 5.04 74.67 0.67 ± 0.53 1–2 ±57 608.2 ± 38.7

15–31 19–35 0.94 ± 0.18 18.50 ± 4.18 82.67 0.76 ± 0.23 1–2 ±50 534.1 ± 42.7

15–30 19–35 0.91 ± 0.15 18.5 0 ± 4.18 82.67 0.75 ± 0.50 1–2 ±50 465.3 ± 41.9

Table 7 Comparison of growth and reproduction of P. ceylanensis with other vermicomposting earthworms. Earthworm species

Medium used

Growth rate (mg/worm/day)

Maximum individual weight (mg/worm)

Fecundity (cocoon/ worm/week)

Hatching success (%)

Reference

Perionyx excavatus

Cattle manure Cowdung

3.5 –

600.0 –

7.70 2.94

72.0 52.5

Cowshed manure

6.7a

608.6

1.79

54.8

Cattle waste solid

6.6a

612.6

1.38

75.9

Reinecke et al. (1992) Bhattacharjee and Chaudhuri (2002) Suthar and Ram (2008) Suthar (2009)

Cowshed manure

7.4a

614.7

–

54.6

Cattle waste solid

a

8.0

767.7

1.58

68.6

Cowdung Cowdung

1.8 1.6

608.2 534.1

5.95 6.58

74.7 82.7

Cowdung

1.3

465.3

6.37

82.7

Perionyx sansibaricus

Perionyx ceylanensis Worms cultured singly Worms cultured in batches of four Worms cultured in batches of eight a

Values are converted from the original values. i.e., from mg/worm/week to mg/worm/day for better comparison.

Suthar and Ram (2008) Suthar (2009) Present study


4796


ceylanensis, and hence it is essential to maintain the optimum number while mass culturing these worms. Bhattacharjee and Chaudhuri (2002) reported that high rate of cocoon production coupled with short incubation period in P. excavatus, Dichogaster modigliani, Pontoscolex corethrurus and L. mauritii are indicative of their potential for utilization in vermiculture based biotechnology. The greater the earthworm population density in a culture, the slower was the growth of individual earthworms at any particular earthworm population density, up to a maximum of 16 earthworms in 100 g of separated cattle waste solids for E. eugeniae (Dominguez et al., 2001). Vermicomposting studies (120 days) conducted to optimize the number of worms required for efficient organic substrates conversion based on the reduction of C/N ratio, percentage decomposition, total number and biomass of earthworms introduced and recovered from the vermibeds clearly showed that vermibeds with 4 kg of organic materials can hold about 60–80 L. mauritii and about 90–120 P. ceylanensis for efficient decomposition (Karmegam and Daniel 2008b). P. ceylanensis produced viable cocoons which hatched parthenogenetically. This indicates that copulation is not essential for cocoon production in P. ceylanensis. The average incubation period of the cocoons ranged from 18.5 to 19 days for P. ceylanensis. The incubation period of the cocoons of P. excavatus was reported as ±18 days. In the present study it was ±18.5 days for P. ceylanensis cultured in batches of four and of eight and ±19 days for singly cultured worms. Edwards (2004) reported that P. excavatus and P. hawayana produced 24 cocoons (13 young worms) and 10 cocoons (9.5 young worms) per week per parent earthworm, respectively. The growth and clitellum development was 28 days for P. excavatus (Hallatt et al., 1990). In the present study, it was 26, 22 and 22 for P. ceylanensis cultured singly, in batches of four and of eight, respectively. A close similarity is observed in the pattern of development and reproduction between P. excavatus and P. ceylanensis. In the present study, hatching success was recorded for P. ceylanensis ranged between 74.7% and 82.7%. Where as, the hatching success reported for other Perionyx species, i.e., P. excavatus and P. sansibaricus are lower which ranged between 52.5% and 72.0% (Table 7). The majority of the cocoons produced one hatchling in P. ceylanensis. Occasionally two hatchlings were produced. 4. Conclusion The growth (avg. growth rate: 1.34–1.79 mg/worm/day), reproduction (0.85–0.94 cocoons/worm/day with the hatching success of 74.67–82.67%) and life cycle (±50 days) of P. ceylanensis seems to be on par with the vermicomposting worms like P. excavatus and P. sansibaricus. Since the duration of life-cycle and the period of incubation are lesser, P. ceylanensis exhibits favorable features for adoption as vermicomposting worm. Acknowledgements Authors sincerely thank the Department of Biotechnology (DBT), Government of India, New Delhi, for financial support (BT/ PRI 141/SPD/16/682/98); Dr. J.M. Julka, Solan, Himachal Pradesh, for the identification of the earthworm (Ref.: EI/3 dt. 12-03-1999 and 22-01-2000); Gandhigram Rural University for granting permission to publish part of the Ph.D. thesis of Dr. N. Karmegam.

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