JOURNAL OF SCIENCE OF HNUE Natural Sci., 2011, Vol. 56, No. 7, pp

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1 JOURNAL OF SCIENCE OF HNUE Natural Sci., 2011, Vol. 56, No. 7, pp INITIAL RESULT OF THE RAISING DI LONG (EARTHWORMS) (PHERETIMA ASPERGILLUM PERRIER, 1872) Tran Thi Thanh Binh ( ) Hanoi National University of Education Nguyen Thi Hien Phu Lo High School, Soc Son District, Hanoi ( ) binhttt@hnue.edu.vn Abstract. This paper provides some initial information about raising Di Long (earthworms) (Pheretima aspergillum). Di Long was fed by 3 different kinds of foods: 100% fresh cow manure (formula 1), 100% cow manure (composted) (formula 2), 70% cow manure (composted) + 30% banana trunk (composted) (formula 3), food was provided fully for 10 days (and their mass was recorded). result of this research showed that the mature Di Long (Pherrtima aspergillum) which has ability to survive, growth and breed in cultured conditions. During experimenting, their mass reached the highest point after months. After 3 months culture, the breeding coefficient was low (r = 0.38), the number of individuals increased (138%) in comparison with that at the beginning and in the immature nextgeneration. After 6 months culture, the breeding coefficient (r) was 1.5, the number of individuals increased (250%) in comparison with that at the beginning and in the mature next-generation. Fresh cow manure provided the highest nutrition for Di Long. refore, the rate of breeding, the number of individuals and their mass were the highest in three nutrient formulae. In addition, by using this formula the amount of food spending for 1 kg increasing Di Long s mass was at its lowest. 1. Introduction Earthworm has been used to decompose organic wastes, provide protein for animal husbandry, improving impoverished soil, environmental indicator for many years [1]. In recent years, with the development in modern medicine and biotechnology humans have extracted some enzymes from earthworms in order to cure some diseases. For example Mihara et al. isolated a protease enzyme group which hydrolyzes fibrin fibers and dissolve blood clots from Lumbricus rubellus [7]. After that, scientists discovered many enzymes in this group from many different species 144

2 Initial result of the raising Di Long (earthworms) (Pheretima aspergillum Perrier, 1872) [5, 6]. Nguyen Thi Ngoc Dao also extracted fibrinolytic enzyme from earthworm(perionyx excavatus) that treat brain strokes. Also, people have utilized many different earthworm species (Perionyx excavatus, Eisenia fetida, Eisenia andrei, Pheretima asiatica, etc.) in order to provide protein for husbandry [3]. Di Long (Earthworms) (Pheretima aspergillum) are distributed widely in the north and central of Vietnam, China, Taiwan, the Island of Madagascar[2, 4]. In folk medicine, Di Long (earthworms) are used for treating some diseases such as malaria, typhoid, epilepsy, the plague. Nowadays, they are being used to make up a medicine - Doragon, which can quickly reduce itching, dry wounds, increase adaptability of the body in strenuous working conditions, is produced by pharmaceutical company Domesco [8]. Di Long (earthworms) are being studied in order to treat primary hypertension and there are initial successful results in their study. Although Di Long (earthworms) have many useful functions, there is no research about worm s breeding and application. In this paper, we provide some initial information about experimenting on multiplying Di Long (Pheretima aspergillum). 2. Content 2.1. Material, period and methods * Material and of study This study has been conducted from 06/2011 to 06/2012, divided into two periods: - Period 1 (06/ /2010): examined survival ability and growth level in cultures. This experiment was conducted with four repeats. Six individual earthworms were raised in a box (20 cm 40 cm 20 cm). Every ten days, we checked and recorded the result. This period was conducted in the laboratory of the Animal Department, School of Biology, Hanoi National University of Education. - Period 2 (01/10/ / 06/2011): examined growth level in different foods. This experiment was repeated in 2 periods (10/ /2010) and (01/ /2011). mature Di Long (Pheretima aspergillum) and soil were used for this experiment which was collected in nature. Soiled was smashed and dried from 2 to 3 days before it was humidified by water spraying. n it was used in the experiment. * Methods This experiment was conducted in cells which were built by bricks and cement ground (1 m 0.5 m 0.5 m). Every cell has a water-outlet in the bottom and an anti-ant slot around the cell (10 cm). Every cell has 50 adult earthworms(pheretima aspergillum) which were recorded in their masses. This step was repeated after the experiment was completed. 145

3 Tran Thi Thanh Binh and Nguyen Thi Hien purpose of this step is to check the earthworm s quantity and mass. DiLong was fedby 3different kinds of foods: 100%fresh cow manure (formula 1 or F1), 100% cow manure (composted) (F2), 70% cow manure (composted after 2 months) + 30% banana trunk (composted after 2 months) (F3), food was provided fully in 10 days (their mass recorded). Some quotas were being recorded during experimental progress such as: increasing number of Di Long (individuals) are the number of Di Long after completing this experiment (individuals) subtracting the number of Di Long at the beginning (individuals) increasing Di Long s mass (g) are the Di Long s mass after completing the experiment (g) subtracting their mass at the beginning (g). Earthworms growth coefficient (%): r = dn Nt N with dn =, where N Ndt t is the number of earthworm at the beginning and t is. M = M 1 M 2, where M: amount of food spending for 1 kg increasing earthworm s mass (kg); M 1 : sum of food being used; M 2 : Increasing earthworms mass (kg). This period was carried out at Nguyen Thi Hien s household, Noi Phat Hamlet, Phu Lo Commune, Soc Son District, Hanoi City Results and discussion survival rate and breeding ability of Pheretima aspergillum in cultured conditions Survival rate: With 4 cells, each cell cultured 6 mature individuals. After 2 months, 6 adults were collected. Thus, the survival rate of Di Long after 2 months is 100% in culture condition (Table 1). This shows Di Long has adapted to culture conditions; therefore, this species can be cultured. Table 1. survival rate of Di Long in cultured conditions Cell 1 Cell 2 Cell 3 Cell 4 number of Di Long at the beginning 6C 6C 6C 6C number of Di Long after 2 6C 6C 6C 6C months survival rate 100% 100% 100% 100% Note. C: Mature individual 146

4 Initial result of the raising Di Long (earthworms) (Pheretima aspergillum Perrier, 1872) Breeding ability: re were no earthworm follicles during experimental although they were in an adult state. Perhaps the reason is the disorder after checking mass or high density which is not suitable with their breeding habits. Growth ability: although individual worms were tested, the adult ones (sexual belt), their mass has increased. This rise was still repeated after 3-4 s but it reached a balanced status or somes decreased in the next test. average mass is g/individual (the biggest one is 27.8 g) which compares with the average mass at the beginning is g/individual. After 1.5 months, the worms mass increased 1.48 s. result shows worms average growth coefficient rose after every test, the highest one was the fourth test which reached % but their tendency slightly decreased (Table 2). Hence, Di Long Pheretima aspergillum is able to survive and grow although the samples were all mature. Table 2. Di Long s growth ability in cultured conditions At the beginning 1st 2nd 3rd 4th 5th 6th Di Long s average mass (g) Growth rate (%) ± ± ± ± ± Di Long s growth ability in different kinds of nutrition ± 6.85 * Period 1 (3 months) (10/ /2010) Breeding and growth ability in culture condition is illustrated in Table 3. Breeding ability: result of this research has shown that Di Long s breeding coefficient after 3 months in all nutrient formulae was larger than 0 (F1: r = 0.24; F2: r = 0.38 and F3: r = 0.30). breeding coefficient after 3 months showed that Di long is able to breed but the breeding rate is rather low. In this nutrient formula (F1), the number of individuals increased to 12 and they were immature (no sexual belt). In F2 the number of individuals increased to 19 which were all immature worms. In F3 that number increased to 15 immature ones. Hence, 3 months in cultured conditions was enough for Di Long to become mature although they had reproduced. Growth ability and the amount of food spending for 1kg increasing Di Long s mass: Di Long grew best in fresh cow manure (F1) both increasing mass and growth rates (313.1g and % respectively), followed by in F2 (261.2 g and %) and the lowest in F3 (243.1 g and %). It can be seen that fresh cow manure is the most suitable food to grow Di Long. amount of food spending for 1 kg 147

5 Tran Thi Thanh Binh and Nguyen Thi Hien increasing earthworms mass increased, which was at the highest in F2 ( kg), followed by F3 ( kg), and the lowest was in F1 ( kg). refore, it can be shown that fresh cow manure is the best nutrition for culturing Di Long. Quotas number of earthworms at the beginning number of earthworms after experimenting number of increasing Di Long Breeding coefficient (r) Di Long s mass at the beginning (g) Di Long s mass after experimenting (g) Increasing earthworm s mass (g) Table 3. result of culturing Di Long after 3 months and 6 months Kinds of nutrition Period 1 (3 months) Period 2 (6 months) F1 F 2 F 3 F 1 F 2 F 3 50 C 50 C 50 C 50 C 50 C 50 C (50C + 12A) (50C + 19A) (50C + 15A) 12A 19A 15A (80C + 45A) 30C + 45A (72C + 38A) 22C + 38A (73C + 52A) 23C + 52A Growth rate (%) quantity of food (kg) amount of food spending for 1 kg increasing Di Long s mass (kg) Note. C: Mature individual; A: Immature individual; F1: Fresh cow manure; F2: Cow manure (composted after 2 months); F3: (70% Cow manure + 30% banana trunk) (composted after 2 months) *Period 2 (6 months) (1/01/ /06/2012) It is expected that earthworms will be repeated twice from January, 2011 to June, However, three months in culture condition they reproduced but was 148

6 Initial result of the raising Di Long (earthworms) (Pheretima aspergillum Perrier, 1872) not enough for earthworm to be mature. Thus, we experimented the second batch for six months. Breeding ability: result of Di Long s breeding ability (Table 3) shows that Di Long s breeding coefficient was larger than 0 in all nutrient formulae after 6 months. (F1: r = 1.5; F2: r = 1.2 and F3: r = 1.5). breeding coefficient and breeding rate this were larger than the experiment with 3 months (F1, F2, F3 and 6.25 s, 3.16 s, 5.00 s, respectively). With 50 mature individuals at the beginning, the number of Di Long was collected after 6 months is shown as follows: In100%freshcowmanure(F1): thenumberofwormswascollectedis125individuals which increased 250% compared with the number at the beginning. Specifically, the number of mature individuals were collected is 80 (160% compared to those at the beginning) and those of mature ones was 45. In 100% fresh cow manure (composted after 2 months (F2): the number of worms was collected is 110 which increased 220% compared with the number at the beginning. Specifically, the number of adults is 72 (144%), which increased 22 individuals compared to the number at the beginning. re were 38 worms which did not have sexual belts. In F3 (70% cow manure + 30% banana trunk (composted after 2 months)) the number of Di Long was collected 125 individuals which increased 250% compared to that number at the beginning. Specifically, the number of adults is 73 (146%), which increased 23 individuals; there were 52 individuals which did not have sexual belts. Comparing the breeding coefficient and growth rate of Di Long shows that the breeding coefficient is equal between F1 and F3 (r = 1.5). However, the number of adults were collected highest in F1 (80 adults/125 individuals), followed by in F3 (73 adults/125 individuals) and the lowest was in F2 (72 adults/110 individuals). Hence, after 6 months Di Long has bred and the next-generation developed until mature. 250% is the highest percentage of the increasing number whereas 160% is the highest percentage of increasing adults specifically which are compared with those numbers at the beginning. breeding coefficient (r) reached 1.5. Growth ability and the amount food spending: Increasing earthworm s mass and earthworms growth rates were different among three kinds of cultures. Specifically, mass and growth rates were highest in F1 (971.2 g, %), followed by in F3 (787.8 g, %) and lowest in F2 (766.8 g, %). According to the result, earthworms still develop well in fresh cow manure which is similar to the first result. 149

7 Tran Thi Thanh Binh and Nguyen Thi Hien number of individuals was collected after 6 months. That shows the two following things. Firstly, it took 6 months to grow earthworms to become adult earthworms. After experimenting, the total number of adult earthworms was the highest in F1 while in F1 (80 individuals) and F3 had the same number of individuals, i.e between 72 and 73 individuals each. amount of food spending for 1 kg increasing earthworms mass which is the lowest in F1 ( kg), followed by in F2 and F3 ( kg and kg respectively). result shows that the level of food spending for the second batch is lower than the first batch in all three kinds of cultures (Figure 1). This complies with the law: faster earthworms growth rate is, the lower the level of food spending for earthworms is. In Dang Vu Binh and et al. s study, the food spending for 1kg increasing earthworms (Perionyx excavatus) reached the highest point at kg [4]. Thus, the food spending for 1 kg increasing earthworms mass in this experiment is larger than Perionyx excavatus. Figure 1. amount of food spending for 1 kg increasing Di Long s mass 3. Conclusion After one year of studying and experimenting on raising Di Long, we obtained the following initial result: - mature Di Long (Pherrtima aspergillum) had the ability to survive, grow and breed in cultured conditions. - mature Di Long was captured in nature still increased their mass during experimenting. Particularly, their mass reached the highest point after months. After 3 months culture, the breeding coefficient was low (r = 0.38), the number of individuals increased (138%) in comparison with that at the beginning and in the immature generation. After 6 months culture, the breeding coefficient (r) was 1.5; the number of individuals increased (250%) in comparison with that at the beginning and in the mature next-generation. 150

8 Initial result of the raising Di Long (earthworms) (Pheretima aspergillum Perrier, 1872) - Fresh cow manure provided the highest nutrition for Di Long; therefore, the rate of breeding, the number of individuals and their mass were the highest in three nutrient formulae. In addition, by using this formula the amount of food spending for 1 kg increasing Di Long s mass was at its lowest. REFERENCES [1] Thai Tran Bai, Value in Application of Earthworms. Journal of Biology, 3, pp (in Vietnamese). [2] Thai Tran Bai, diversity of Earthworms in Vietnam. Workshop proceedings of the basic problems in biological research. Vietnam National University Publishing House, Hanoi, pp (in Vietnamese). [3] Dang Vu Binh, Vu Dinh Ton, Nguyen Dinh Linh Eluavation of Earthworm (Perionyx excavatus) Growth Raised in Different Cultures. Journal of Science and Development, VI (4), pp (in Vietnamese). [4] Blakemore R. J., Cosmopolitan Earthworm-an Eco-Taxonomic Guide to the Peregrine Species of the World. Published by VermEcology, PO BOX 414 Kippax, ACT 2615, Australia, pp [5] ChoH., ChoiE.S., LimH.G.,LeeH.H.,2004.Purification and Characterization of six Fibrinolytic Serine - Proteases from Earthworm Lumbricus rubellus. Journal of Biochemistry and Molecular Biology, 37(2), pp [6] Lu Y., Jin R., WuY., Mang X., purification and characterization of fibinolytic enzymees from Amynthas dancatala. Chinese Journal of Biochemistry & Molecular Biology, 4, pp [7] Mihara H., Sumi H., Akazawa K., Fibrinolytic enzymee extracted from the earthworm. Thrombosis and Haemostasis, 50, 258. [8] 151