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Article# Article Title & Authors (Volume 6, No. 2, April 2017) Page
1 Preparing Silk Biomaterial from Cocoon of Wild Silkmoth Attacus atlasTjokorda Sari Nindhia, Tjokorda Gde Tirta Nindhia, I Wayan Surata, Zdenek Knejzlik, and Tomas Ruml
32
2 Development of Water in Olive Oil (W/O) Nanoemulsions as Lipstick Base FormulationSiti Z. Munawiroh, Aughina N. Nabila, and Lutfi Chabib
37
3 Physicochemical Properties of Spray-Dried Young Coconut JuicePimolmart Rattanaburee, Thanaporn Amnuaikit, and Jindaporn Puripattanavong
43
4 Correlation of Balance Ability and Physical ConditionsYi-Yang Chen, Xu-Yang Gao, Li-Chi Liao, Chia-Ling Lee, and Chaur-Heh Hsieh
48
5Selection of Oleaginous Yeasts with Lipid Accumulation by the Measurement of Sudan Black B forBenefits of BiodieselKusumawadee Thancharoen, Auttaporn Malasri, Walailak Leamsingkorn, and Patchareeporn Boonyalit
53
6Predicting the Effect of Parathyroidectomy on Patient Survival in Secondary Hyperparathyroidism withMachine LearningOktoria, Cheng-Hong Yang, and Jin-Bor Chen
58
7 The Effect of Orchis anatolica Leaves on Reproductive System—An in Vivo StudyMansour M. Nawasreh and Lubna H.Tahtamouni
63
8 Acetylcholinesterase Inhibitory and Antioxidant Properties of Thai VegetablesSupat Langyanai, Prapaporn Chaniad, and Jindaporn Puripattanavong
67
9A Study of Proliferative Activity of Herbs Eriocaulon Cinereum R.Br on Cervical Cancer Cells (HeLa) withMTT Assay MethodArde T. Nugraha, Viren Ramadhan, Herianto Pandapotan, and Fitra Romadhonsyah
73
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Preparing Silk Biomaterial from Cocoon of Wild
Silkmoth Attacus atlas
Tjokorda Sari Nindhia Faculty of Veterinary Medicine Udayana University, Bali, Indonesia, 80000
Email: [email protected]
Tjokorda Gde Tirta Nindhia, and I Wayan Surata Mechanical Engineering, Udayana University, Jimbaran, Bali, Indonesia, 80361
Email: [email protected], [email protected], [email protected]
Zdenek Knejzlik and Tomas Ruml Faculty of Food and Biochemical Technology, Technicka 5, 16628, Prague Czech Republic
E-mail: {zdenek.knejzlik, tomas.ruml}@vscht.cz
Abstract—This Article reporting result from research in
providing silk from wild silkmoth cocoon of Attacus atlas.
The silkmoth is from Indonesia origin. The research was
initiated by collecting the silkmoth egg from their real
location. The silkmoth eggs were hatched and the larva were
introduce with specific leaf that will be use as a a future food.
The consideration in selecting leaf for the food is the leaf
should from the three that easily and rapid grow also having
ingredient that could be useful in drug, pharmacy and
medical purpose. It is found the wild silkmoth Attacus atlas
are like to consume a herb plant leaf of Erythrina variegate
and also Cananga odorata. The cocoon resulting from
feeding the caterpillar of Attacus atlas with these 2 type of
leaves were observed by using scanning electron microscope
(SEM) and element compositions were detected by using
Energy Dispersive X-ray Spectroscopy (EDS). Different
appearance of fiber are observed. With feeding with
Cananga odroata resulting fiber surface covering with cubic
crystal that reach with Calcium (Ca) meanwhile feeding
with Erythrina variegate resulting fiber without fibrous
surface which is rich with clhorine (Cl).
Index Terms—silk, wild, cocoon, Attacus atlas, Cananga
odorata, Erythrina variegate, calcium, chlorine
I. INTRODUCTION
Silks that is obtained from the cocoon are become
long-standing interest by the scientist. The scientist put
attention on appearance and behavior of cocoon produced
by different type of silkworm. Recently silk have been
developed as engineering fibre to make artificial
composite for biomaterial in medical. To make it
constructive and effective, the cocoon structure
biophysics should be understood [1].
The mechanical properties of silk are impressive, with
slow degradability and biocompatibility that render silk
as a biomaterial for further exploration. Tissues such as
bone, tendons, or ligaments are incapable of self-repair
Manuscript received February 6, 2017; revised April 20, 2017.
because of the damaged or diseased. It is often required a
substitute biomaterial to aid the healing process. silk
fibroin protein is mechanically robust, can be designed to
desired specifications, and biocompatible [2].
Silks are protein that are spun into fibers by larvae
such of mites, silkworms, scorpions, spiders and flies
(Lepidoptera). Silk are produced in glands through
biosynthesis process in epithelial cells. The proteins are
stored into the lumen of these gland prior spinning into
fibers. The most characterized silks are from the
domesticated silkworm, Bombyx mori, Each silks has a
different composition of amino and exhibits mechanical
properties to their specific functions [3]
The silkmoth Attacus atlas (Lepidoptera; Saturniidae),
is an insect species that can produce silk with having
value of ornamental. This moth can have maximum wing
span to reach 300 mm and is the largest wing of all moths
(Fig. 1). This species is found in china and Southeast
Asia in tropical rainforest habitats at altitudes around
1500 m. Not like domesticated mulberry silkworm of
Bomyx mori that eat only one type of plant (mulberry
plant), the larvae or Attacus atlas feed variety of plants
They often move from one plant to another in their
development [4].
The larvae of Attacus atlas produce cuticular wax from
abdominal tubercles. This wax forms a white, dense,
powder coating over lateral surfaces and dorsal of the
larva. The function of waxes is the conservation of water,
anti parasitoid, anti predator. The larvae spin silk threads
to form a cocoon. The function of the cocoon is to
enwrap for metamorphosis. The cocoon are composed of
two proteins namely, sericin and fibroin. The functional
of these protein structures includes barricade from
predators, parasitoids, pathogens and assisting to
complete metamorphosis by moisture and thermal. The
variations in diet, and climate resulted in diversity in the
type and properties of the cocoons and fibers produced by
silk within the same type [5].
International Journal of Pharma Medicine and Biological Sciences Vol. 6, No. 2, April 2017
©2017 Int. J. Pharm. Med. Biol. Sci. 32
doi: 10.18178/ijpmbs.6.2.32-36
Plants derived biologically active compounds have
become important source of drug. Because of the
increasing recognition of herbal medicine as an
alternative of health care. Erythrina variegate plants
contain phyto pharmaceuticals, which have very
important applications in the fields of medicine for the
prevention of diseases. Medicinal plants provide
molecules that lead to the discovery of new drugs. Plant
metabolites that are phenolic exhibit antiallergenic,
antimicrobial, antiatherogenic, antithrombotic,
antiinflammatory, cardioprotective and vasodilatory
effects [6].
Figure 1. Silkmoth and cocoon of Attacus atlas, Indonesia origin.
Traditionally the bark of Erythrina variegate is used to
treat swellings and stomachache with other herbs. Its
leaves are occasionally used to treat eye ailments, to
stimulate lactation and menstruation for women, to treat
wind-damp obstruction syndrome in rheumatic joint pain
and spasm of the limbs or lower back and knee pain.
Phytochemical studies on Erythrina variegate plant have
led to the isolation of many isoflavones. Isoflavones are
compounds in plant foods, structurally similar to the
mammalian estrogens. Recently received ttention for
potential use in the prevention of postmenopausal bone
loss. Data from animal experiments provided evidence
that isoflavones can attenuate menopausal bone loss, and
it was suggested that isoflavones is responsible for
protective effects on bone [7].
It was informed that Erythrina variegate has been used
in folk medicine for treatment of a narcotic and
antihelminthic, tooth-ache, veneral disease, asthma,
malarial fever, and insomnia. The alkaloid Erythroidine
from Erythrina variegate was used as a muscle relaxant
as well as Haemoerythrina alkaloids from Erythrina
variegate were investigated for their anti-cancer activity
[8].
Erythrina variegata is found in many tropical and
subtropical regions. Phytochemical investigation of the
non-alkaloidal secondary metabolites revealed the
presence of one cinnamyl phenol and several
isoflavonoids which exhibit activities of antibacterial and
anti-inflammatory. Erythrina variegata is used
medicinally as an antibacterial, antipyretic, anti-
inflammatory, antiseptic agent and as a collyrium [9].
Erythrina variegate having positive result as antifungal
again Pythium ultimum, Rhizoctonia solani, Aspergillus
fumigates, but negative for Phytophthora parasitica. It is
also found that Erythrina variegate having negative result
as antibacterial for Escherichia coli, Staphylococcus
aureus [10].
Cananga odorata grows natively in South-East Asia,
Cananga odorata is a perennial tropical tree and reported
has a variety traditional uses and medicinal properties
Cananga odorata oil is used to enhance euphoria during
sex and also reduce sexual anxiety. The leaves of
Cananga odorata is can relieve itchiness by direct topical
application and also to treat dandruff [11].
It was reported that ethanolic extract of the root bark of
Cananga odorata causes spermatotoxicity in male albino
rats, suppresses the production of testosterone, and alters
the micronutrients of the testis [12]. The bark of Cananga
odorata reveal as antibacterial and antifungal activity
[13].
Cananga odorata has been long utilization as
fragrance component in perfumery and increase in recent
year for flavoring agent in chewing gums, candies, frozen
dairy, icings, candies, baked goods and soft drinks. the
consumption of cananga oil from food flavoring does not
pose human health effects. The Cananga odorata oils
have been determined to be safe (GRAS) for food uses by
the International Organisation of Flavor Industries (IOFIs)
and Flavor and Extract Manufacturers Association
(FEMA) and . Have been approved as food additives by
US Food and Drug Administration (FDA) [14].
In this research, the leaves from Erythrina variegate
and Cananga odorata plant are manage to become a food
for Attacus atlas larvae. The cocoon obtained from each
plant leaf will be observed and the element contain in it
will be identified.
II. EXPERIMENTAL
The leaf of Erythrina variegate (Fig. 2) and Cananga
odorata (Fig. 3) were prepared from Indonesian source.
The egg of Attacus atlas were collected from their
original location in Indonesia. That was provided 2
groups of larvae, one with only eat leaf of Erythrina
variegate (Fig. 4) and the other group just eat leaf of
Cananga odorata (Fig. 5)
Figure 2. Leaf of Erythrina variegate.
The caterpillars of Attacus atlas were let to consume
the leaves prepared and in final stage the silkworm
provide themselves a cocoon for final step of
metamorphose. The silkmoth of Attacus atlas was let to
exit from the cocoon for the next breeding. The cocoons
left by the silkmoth were collected and drying in the oven.
The microstructure of the virgin cocoon were investigated
International Journal of Pharma Medicine and Biological Sciences Vol. 6, No. 2, April 2017
©2017 Int. J. Pharm. Med. Biol. Sci. 33
and recorded under scanning electron microscope (SEM)
with different magnification. To reveal of elements
compositions in the cocoon, the Energy Dispersive X-ray
spectroscopy method was introduced. The result was
presented in table and graphic.
Figure 3. Leaf of Cananga odorata.
Figure 4. Carterpilar of Attacus atlas is consuming leaf of Erythrina
variegate.
Figure 5. Caterpillars of Attacus atlas are consuming leaf of Cananga
odorata.
III. RESULT AND DISCUSSION
The cocoon that obtained is presented in Fig.6 for
feeding with leaf of Erythrina variegate as well as in Fig.
7 for the one that feeding with Cananga odorata. The
cocoons were found about in the same size and give
explanation even though the silkmoths consume 2
differents2 type of the leaves, they giving the same size
of cocoon. This is also an indication that composition of
the leaves of Erythrina variegate and Cananga odorata
having about the same amount nutrient. This is also
answering the question about why they like to consume
these 2 types of leaves.
Result on observation by using SEM with low
magnification (100x) can be seen at Fig. 8 for Erythrina
variegate and Fig. 9 for Cananga odorata. At low
magnification, the physical appearance the both cocoon is
not so much different. The different between 2 type of the
cocoon can be clear observed on very high magnification
(10000x). Irregular crystals were found on the surface of
the cocoon that was feeding with Erythrina variegate as
can be seen in Fig. 10. On the other hand by feeding with
Cananga odorata, the surface of the cocoon are found
covering with cubic crystal as can be observed in Fig. 11.
Figure 6. Cocoon of Attacus atlas yield from feeding with Erythrina
variegate.
Figure 7. Cocoon of Attacus atlas yield from feeding with Cananga
odorata.
Figure 8. Cocoon of Attacus atlas yield from feeding with Erythrina
variegate (100x).
International Journal of Pharma Medicine and Biological Sciences Vol. 6, No. 2, April 2017
©2017 Int. J. Pharm. Med. Biol. Sci. 34
Figure 9. Cocoon of Attacus atlas yield from feeding with Cananga
odorata (100x).
Figure 10. Irregular crystal shape were found on the surface of the
cocoon feeding with Erythrina variegate (10000x).
Figure 11. Cubic crystals were found on the surface of the cocoon
feeding with Cananga odorata (10000x).
Elemental analysis with energy dispersive X-ray
sprectroscopy (EDS) on the surface of the cocoon reveal
that by feeding the larvae of Attacus atlas with Erythrina
variegate resulting the cocoon contain chlorine (Cl)
around 2.96 mass% (Fig. 12). Chlorine is not found oin
the cocoon that was feeding with Cananga odorata but in
reply the element of Calcium (Ca) is present around
14.37 wt%. The presence of calcium in the cocoon feed
with Cananga odorata will give good result in
biocompatibility since calcium is essential for cell
physiology [12]. The other elements were found the same
that are carbon (C), oxigent (O), and kalium (K) as can be
seen at Table I and Table II. Similar result was found for
other species of wild silk that having cubic crystal on the
surface such as Hyalophora gloveri , Bunaea aleinoc and
Antheraea pernyi [1].
Figure 12. Energy dispersive X-ray sprectroscopy on the cocoon
feeding with Erythrina variegate.
TABLE I. ELEMENTS COMPOSITION IN THE COCOON FEEDING WITH
ERYTHRINA VARIEGATATE
No Element Mass (%)
1 C 64.86
2 O 26.27
3 Cl 2.96
4 K 5.92
Figure 13. Energy dispersive X-ray sprectroscopy on the cocoon
feeding with Cananga odorata.
TABLE II. ELEMENTS COMPOSITION IN THE COCOON FEEDING WITH
CANANGA ODORATA
No Element Mass (%)
1 C 33.49
2 O 49.49
3 K 2.29
4 Ca 14.73
IV. CONCLUSION
Wild silkworm cocoon of Attacus atlas is possible for
feeding with leaves of Erythrina variegate and Cananga
odorata. The surface of the cocoon is covered with
irregular crystal shape that is rich with chorine (Cl).
Feeding the silkworm Attacus atlas with Cananga
odorata resulting cocoon that is rich with calcium (Ca)
with cubic crystal shape
ACKNOWLEDGMENT
The authors wish to acknowledge financially support
from grants of the non-tax revenues (PNBP) from the
scheme of international collaboration research (PNBP) of
Udayana University, Bali, Indonesia with collaboration
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International Journal of Pharma Medicine and Biological Sciences Vol. 6, No. 2, April 2017
©2017 Int. J. Pharm. Med. Biol. Sci. 35
with the Institute Chemical Technology Prague, Czech
Republic.
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Tjokorda Sari Nindhia is Doctor in
Veterinary Medicine (DVM) at faculty of
Veterinary Medicine Udayana University, Bali,
Indonesia since 1999. Finishing her master
degree at biotechnology and biomolecular at
postgraduate studies Udayana University in
Bali, Indonesia
Tjokorda Gde Tirta Nindhia received
Doctor Degree in Mechanical Engineering
from Gadjah Mada University (UGM)
Yogyakarta, Indonesia on August 2003, with
major field of study was Material Engineering.
His current job is as Full Professor in the field
of Material Engineering at Department of
Mechanical Engineering, Engineering Faculty,
Udayana University, Jimbaran, Bali,
Indonesia. His research interest covers
subjects such as, biomaterial, waste recycle,
failure analyses, ceramic, metallurgy, composite, renewable energy, and
environmental friendly manufacturing.
Tomas Ruml is full professor and dean of the
Faculty of Food and Biochemical Technology,
Technicka 5, 166 28 Prague Czech Republic.
Zdenek Knejzlik. is senior researcher at
Faculty of Food and Biochemical Technology,
Technicka 5, 166 28 Prague Czech Republic.
I Wayan Surata received Doctor Degree in
the field of Ergonomic from Udayana
University in 2011. His research interest very
much related in process of manufacture. His
is a full professor at Department of
Mechanical Engineering, Engineering Faculty,
Udayana University, Jimbaran, Bali,
Indonesia.
International Journal of Pharma Medicine and Biological Sciences Vol. 6, No. 2, April 2017
©2017 Int. J. Pharm. Med. Biol. Sci. 36
