An Attempt Of Atakas Wild Silkmoth Biology Essay

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Evaluation of atlas moths, Attacus atlas, artificial diet nutrition through haemolymph protein quantification was investigated. Four species of different plant fodders used as the basis component in the artificial diets, namely Barringtonia, mahogany, Nauclea and soursop. Nutritional value evaluation was done based on the analysis of haemolymph proteins of 6th instar larvae using Folin Lowry procedure.

The results of all parameters showed that the survivorship, cocoon quality, and haemolymph protein content of those larvae fed on barringtonia-based artificial diet was better than of those larvae fed on mahogany, Nauclea, and soursop based artificial diets. The average adults emergence was 74.53 %. The cocoon and empty cocoon yield weights average was 7.01 and 1.1 g, respectively. The protein contents of larvae those fed on barringtonia-based artificial diet show the highest with an average of 28.06 mg/ml. The rearing of atlas moth on barringtonia-based artificial diet was comparable to those of natural feeds, although still need further refinement by adding devices to adjust the humidity, temperature and light intensity.

Key words :Attacus atlas (Linn.), plant fodders-based artificial diets, haemolymph proteins.


An atlas moth, Attacus atlas (Linn.), is known as one of silk-producing insects. The geographic distribution of the atlas moth is in Asia Pacific including Indonesia (Peigler 1989). At the larval stage, this insect is known as a defoliator pest of cinchona, tea, avocado and mango (Kalshoven 1981). Although this species usually categorized as a pest, the quality of the silk is better than those of mulberry silk, Bombyx mori L. (Akai 2000). It makes the atlas moth has a high economic value. Moreover, the species is potential to be cultivated as a silk producing insect like "tasar" (Antheraea mylitta) and "eri" (Samia Cynthia ricini) silks. Not as a mulberry silk in which has been domesticated since 3,000 BC, it is difficult to be reared indoors. Since 1992, the wild silk has been maintained in the laboratory and semi field in the Entomology Laboratory, Faculty of Biology, Gadjah Mada University, Indonesia.

There were 110 species from 49 families of plants in which the atlas moth might be found (Peigler 1989).The cultivation of the wild silk is depending on the insect's diets or hosts, methods of the rearing and environmental factors as well. In Yogyakarta Special Region, there are several plants, such as barringtonia (Barringtonia asiatica), Nauclea (Nauclea orientalis), soursop (Annona muricata) and mahogany (Sweetenia mahagony), which are having high potential to be used as wild silk plant fodders (Situmorang 1996). Many wild silk researches using these plant fodders have been done and the results showed that survivorship of those fed on barringtonia leaves were higher than the other hosts (Situmorang 1996, Tjiptoro 1997, Subagyo 2000, Widyarto 2001, Situmorang and Wuliandari 2002). But the mortality rate in outdoor is still high, up to 30 % (Purwanto and Sukirno 2006). In Antheraea, Attacus, Calosamia cynthia, Platysamia cecropia, Saturnia pyri, Bombyx mori and Eriogaster lanestris, it have been known that environmental factors such as photoperiodic, light intensity, temperature and humidity as common factors affecting cocoon quality and diapause duration(Kato and Sakate 1981, Peigler 1989, Kato and Miyata 1994, Harizanis 2004).

Researches on the atlas moth rearing both by using natural plant fodders and artificial diets have been done. These researches showed that cocoon quality yield from the laboratory rearing were lower than those reared in outdoor (Situmorang 1996, 1997). Situmorang (1997) had reared the atlas moth using an artificial diet which was modified from Antheraea mylitta Drury. artificial diets formulated by Akai et al. (1991). In his formula, he changes the arjun leave powder (Terminalia arjuna) with barringtonia and Chlorella with bee pollen.

Silk fibers industries, until now, depend on the cocoon harvested from the wild. Because of habitat degradation and natural enemies, now days, it is difficult to find the atlas moth in a great number. The rearing of wild silk is very important in order to produce high quality and germ free cultures, nutritional study of the insects and maintain continuous atlas moth for its conservation. Hopefully, if this wild silk could be cultivated, it may fulfill the industry demands. Though, we also conserve and overcome the excessive exploitation of the insects.

Nutritional value of diet is the main factor for wild silk survivorship. The quality of the diet should be evaluated continuously in order to develop satisfied cultivation. The ingested nutrients will be used as cell growth and development. A good diet does not only contain high and complete nutrient but it should be easily digested, absorbed and processed through metabolism as well as support the respiration and excretion (Trager 1953, Coudron et al. 2004)

The aim of this research was to establish and to evaluate the nutritional value of artificial diets used for establishing atlas moth silk cultures in the laboratory scale.

Materials and Methods

This research has been done on November 2008 to April 2009 at Entomology Laboratory, Faculty of Biology, Gadjah Mada University, Indonesia. Larvae used were the progeny of insects reared in the laboratory fed on barringtonia for three successive generations.

1. The study of artificial diets effect on the atlas moth survivorship

The artificial diet formulas used in this research were modified from Situmorang formula (Situmorang 1997). Four different dry green leaf powders used were barringtonia, mahogany, Nauclea and soursop. The ingredients of the diets were consist of dry green leaf powder, cellulose powder, corn pollen, low fat soy bean powder, ascorbic acid, chloramphenicol, potassium hydro phosphate, iron phosphate, calcium carbonate, acrylic acid, sorbic acid, vit. B solution, agar, and distillate water.

Each formula was using 150 neonate larvae divided into 10 replicates, in which reared in ten glass petri dishes (d: 9 cm; h: 1.5 cm) and supported will small bamboo sticks (d: 4 mm; l: 80 mm) and pieces of artificial diet. After 2 days old 2nd instar, the larvae transferred into the bamboo box and fed with artificial diet until 4th instar. Two days old 4th instar larvae then transferred individually into the bamboo box and maintained until pupation. The larvae were checked daily to clean the frass and the diet were changed after maximum one week. Seven days after pupation, the cocoon was collected and measured. The collected cocoons then incubated into mesh cages (l: 50 cm; w: 50 cm; h: 75 cm) until emergence. The parameters observed were larval duration, cocooning percentage, cocoon weight, adult emergence, adult wing span, and eggs fecundity. As a control, as many as 50 neonate larvae in each, in the outdoor were maintained on barringtonia trees, while in the laboratory was used barringtonia leaves.

2. The nutritional value evaluation of the diets

To evaluate the artificial diets quality, quantification of haemolymph protein was made by using Folin Lowry methods (Plummer 1971). Haemolymph of two days old 6th instar larvae, with seven replicates per treatment, were isolated and added with 1-phenyl-2-thiourea (PTC) crystal (Magro et al. 2006). The isolated haemolymph was centrifuged under 10,000 rpm for 5 minutes. The isolated haemolymph protein treated with Folin Lowry reagents. The absorbency of protein was measured by spectrophotometer using 595 nm wavelengths. The standard curve was generated using Bovine Albumin Serum (BSA) as a standard solution. As a control, seven larvae of two days old 6th instar maintained on barringtonia leaves were used.

3. Data analysis

The experimental design used in this experiment was completely randomized design (CRD). The analysis of variance of the treatments was generated using SPSS 13 and further analysis was used Duncan's multiple range test (DMRT) at 0.05 significant levels. The estimation for determining the interaction between haemolymph protein content and atlas moth survivorship was use curve expert ver. 1.3 software.

Results and Discussions

1. The study of artificial diets effect on the atlas moth survivorship

There were three grades of the cocoon, good, medium and poor qualities. Cocoon in which compact, light color and thick were categorized as a good quality, while in which the color is not evenly was as a medium. If the cocoon is thin, not compact, with even or uneven color, it was categorized as poor quality. The average cocoon weight in which categorized as a high quality fiber produced by the larvae fed on barringtonia, mahogany, Nauclea and soursop-based artificial diet were 1.1, 1.03, 0.91 and 0.98 g, respectively (Table ). The intermediate quality of cocoon yield from those fed on a soursop - based diet was the lightest (0.78 g), while those fed on mahogany-based was the heaviest (1.13 g).

Cocoon shell weight produced from those fed on baringtonia-based artificial diet was lighter to those fed on barringtonia leaves. Average cocoon shell weight those fed on barringtonia leaves was 1.89 g, or 41.8% heavier than those on the diet. This may due to nutrient in the natural diet was higher and more complete to those in artificial diets.

In the outdoor rearing, by using mahogany as a host plant the weight of the cocoon was 1.35 g (Subagyo, 2000), while on soursop and Nauclea were 1.26 g and 0.8 g, consecutively (Wahyudi, 2000).Rearing of atlas moth using barringtonia-based artificial diet was produce the best grade cocoon than those fed on others diet formula, even though, it was still lower than those on a natural diet.

Table .Quality of cocoon shell weight yields from atlas moth larvae fed on different artificial diet



Mean (SEM)

Mahogany Mean (SEM)


Mean (SEM)


Mean (SEM)


1.1 (0.03) a

1.03 (0.05) a

0.91 (0.04) a

0.98 (0.08) a


1.0 (0.02)ab

1.13 (0.13) b

0.91 (0.08)ab

0.78 (0.03) a


0.79 (0.03) a

0.7 (0.24) a

0.76 (0.03) a

0.78 (0.03) a

Note: numbers in the same rows which is followed by different letter showing significant differences at α 0.05

The diet formula was affecting the survivorshipsof the atlas moth. Percentage of cocoon yield from those fed on barringtonia-based artificial diet was the highest (86.00 %), while those with other formulas were very low 20%; 14.67 % and 6.67 % for Nauclea, mahogany and soursop, respectively (Table ). This cocooning percentage was higher than those fed on Situmorang's artificial diet and barringtonia tree in the field (control), in which the cocooning percentage was 63.4% (Situmorang 1997) and 44%, respectively. Although the larval duration in barringtonia-based diet was about 5 days longer than the previous study, but the cocoon weight was almost 10% heavier. The cocoon weight on the previous formula was 7.23 g in average. It showed that the rearing on the artificial diet has been enhanced to support the larvae to produce a better quality of cocoon.

Rearing of the insect using Nauclea leaves as the diet in the indoor experiment was showed that the cocooning percentage was 30 %. Larval duration and cocoon yield were ranging from 22 - 31 days and 4,79 - 12,92 g, respectively. Each of the female laid eggs up to 280 eggs (Tjiptoro 1997). While on erythrina, the larval duration was 30-31 days and need 61 - 62 for completing its life cycle (Zebua 1997). The larval duration those fed on soursop and barringtonia were 35 and 28 days, consecutively(Situmorang 1996, Santika 1997)).

Table .The survivorship of the atlas moth to yield cocoon, develop as adults, and eggs fecundity



Diet Formula






Female larval duration (days)

37.26 (0.50)ab

40.22 (0.70) b

40.00 (0.87)ab

37.00 (2.89) a


Male larval duration (days)

38.35 (1.42) a

40.35 (1.17) a

40.33 (2.60) a

35.50 (0.58) a


Cocoon yield (%)

86.00 (2.89) c

20.00 (2.43) b

14.7 (1.66) b

6.7 (1.41) a


Cocoon weight (g)

7.81(0.16) c

6.34 (0.28) a

7.12 (0.21)bc

6.97 (0.97)ab


Female pupal duration (d)

24.13 (1.08) a

24.67 (0.65) a

25.00 (0.58) a

25.50 (1.00) a


Male pupal duration (d)

23.29 (1.42) a

23.45 (0.58) a

22.67 (1.20) a

26.50 (2.73) a


Adults (%)

74.53 (2.78) c

16.57 (1.81) b

13.2 (1.37) b

6.10 (1.53) a


Female wing span (cm)

18.98 (0.37) b

15.94 (0.89) a

16.36 (1.00)ab

18.25 (0.73) b


Male wing span (cm)

17.21 (0.29) a

16.83 (0.79) a

15.48 (2.46) a

17.00 ( 0.84) a


Eggs laid

273.5 (8.99) b

174.4 (21.00) a

149.3 (53.03) a



Eggs fertility (%)

87.39 (19.65) b

76.84 (25.27) a

75.82(48.48) a


Note: numbers in the same rows which is followed by different letter showing significant differences at α 0.05

The result of the study showed that survivorship of the atlas moth to develop as an adult between each treatment was different. 74 % of the atlas moth fed on barringtonia-based artificial diet were succeed to develop asthe normal adults in which were indicated by normal wings, no curled or vestigial parts and able to completely emerge from their cocoon.

The femaleadult of which their larvae stadia's were fed leave-based artificial diets produce a large number of eggs (Table 2). Similarly to other parameters, percentage of eggs hatchingin the barringtonia treatment was also the higher than others. The previous experiments on barringtonia tree showed that it produce both high survivorship and high egg fecundity as well as host preference (Purwanto and Garjito 2002, Purwanto et al. 2002).

Based on the results it was shown that barringtonia-based artificial diet was the best artificial diet formula for laboratory rearing of the atlas moth. The rearing of atlas moth on barringtonia-based artificial diet was comparable to those of natural feeds, although still need further refinement by adding devices to adjust the humidity, temperature and light intensity.

3. The evaluation of artificial diet quality

Nutritional evaluation of the artificial diets through quantification of haemolymph protein showed that barringtonia-based diet gave the highest nutritions than those fed on other formulas(Table ). Compared to the larvae fed on barringtonia leaves in which the protein content was 56.91 mg/ml, the differences between those fed on barringtonia-based diet was 49.3 %. This difference was quite similar to the cocoon shell weight difference, in which 41.8% gap, in the same treatment. It was indicated that there was a relation effect between protein contents and cocoon shell weight yield by the atlas moth.

Table .Haemolymph protein contents of the larvae fed on different formulas



Protein content (mg/ml)


Barringtonia-based diet

28.06 (8.52) b


Nauclea-based diet

17.92 (5.90)ab


Mahogany-based diet

16.05 (2.86)ab


Sousop-based diet

3.09 (2.02)a


Barringtonia leaf

56.91 (7.42) c

Note: numbers in the same rows which is followed by different letter showing significant differences at α 0.05

Beside the nutritional values, both the diet's water content and environmental condition have an important role. A suitable diet for an insect is a diet which has similar water content as its natural food. The water content gives diet's texture that is important for metabolism(Kikuchi et al. 1980). (Mathews and Stephen 1999)(Mathews and Stephen 1999)(Mathews and Stephen 1999)(Mathews and Stephen, 1999)The most important environmental conditions to maintain a good nutritional process are temperature and relative humidity. The survivorship of the insects will be high due to the high nutritional value of the diet as well as high relative humidity (> 80 % RH) (Mathews and Stephen 1999). In order to facilitate a proper cocooning on silk moth, light intensity has an important role (Krishnaswami et al. 1972). This the reason why the rearing of this insect is better on outdoor (Situmorang 1996).

4. Correlation of haemolymph protein and atlas moth survivorships.

The analysis indicates that there was a correlation between haemolymph protein variability to the cocoon weight, female larval duration, cocooning percentage, and adult emergence (Table ).

Table . Regression and correlation of haemolymph protein content and atlas moth survivorships







Female larval duration





Y = 35.28 + 0.61 X - 0.02 X2

Male larval duration





Y = 33.3 + 0.78 X - 0.02 X2

Cocooning %





Y = -15.5 + 3.5 X -0.03 X2

Cocoon weight





Y = 6.79 + 0.002 X + 0.0006 X2

Female pupal duration





Y = 25.62 - 0.03 X - 0.0007 X2

Male pupal duration





Y = 27.81 - 0.47 X + 0.01 X2

Adult emergence





Y = 14.3 - 3.2 X + 0.19 X2

Female wing span





Y = 19.59 - 0.48 X + 0.02 X2

Male wing span





Y = 17.48 - 0.19 X + 0.006 X2

Laid eggs





Y = -10.83 + 10.2 X

Eggs hatchability





Y = 59.6 + 0.99 X

Good cocoon weight





Y = 0.99 - 0.01 X +0.0004 X2

Medium cocoon weight





Y = 1,407.56 + 1.04 X3.3/ 1828+ X 3.3

Poor cocoon weight





0.81 - 0.01x +0.0004 X2

Quantification of protein haemolymph also has been done by (Cremonez et al. 1998)on bee and (Magro et al. 2006)on Bracon hebetor fed on artificial diets. By this experiment, it is indicated that quantification of protein haemolymph could be done as an evaluation of the diet quality. It is such a fast method, no much time consuming and unnecessary to kill the insect as well. This method also may be applied to evaluate the nutritional quality of the diet for poultry, cattle and other livestock.


From this research it could be concluded that quantification of protein haemolymph is such a fast effective, efficient and inexpensivemethod for evaluating the nutritional diet quality. It is also found that barringtonia-based artificial diet was the best qualified artificial diet than those mahogany, Nauclea and soursop-based diet.


We are very gratefully acknowledges Mr. Suparmin and Mr. Paimin for the supporting the technical works. Thanks for the Wild Silk Research Station Sawit Sari for providing the host plants.