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TERATOGENIC TEST OF YOUNG PINEAPPLE FRUIT (Ananas comosus)

ON MOUSE FETUS (Mus musculus L.)

Conference Paper · September 2010

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Iriani Setyawati Dwi Ariani Yulihastuti

Udayana University Udayana University
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 TERATOGENIC TEST OF YOUNG PINEAPPLE FRUIT
(Ananas comosus) ON MOUSE FETUS (Mus musculus L.)
RESULT AND DISCUSSION
Effect on Fetus Morphology
Fetal weight and length tended to decrease with the increasing doses. The rate of fetus growth
ABSTRACT and development determine variation size of the young. Disturbances of individual development
This experiment was performed to examine the effects of young pineapple within uterine caused abnormal birth weight. Growth inhibition occurs when agents affect cell
(Ananas comosus) extract on fetus morphology and skeletal development if given proliferation and interactions, reducing the rate of biosynthesis associated with the inhibition of
to the pregnant mouse during organogenesis period. Twenty pregnant mice were synthesis of nucleic acids, proteins, or mucopolisacharides. Hemorrhage was not found in control, but
randomly divided into 4 groups. Extract was given orally by gavage with 0 increases with the increasing doses. Hemorrhage is a discharge of blood from the blood vessels. It
(control), doses 20%, 40%, and 80%. Treatment was given at day 6 to 15 of will accumulate in body tissues. Extract given repeatedly up to a high concentration in blood resulted
gestation. Caesarean section to remove fetus was performed on day 18 of gestation. osmotic imbalance, caused by osmotic pressure and viscosity of the fluid disturbances in different
The observation covered morphological of the fetus (weight and length of the litter, parts of embryo, between blood plasma and extra-capillary space or between extra-and intra-
and malformations), skeletal development (ossification number of metacarpus, embryonic fluids. It will rupture blood vessels and cause hemorrhage (Price and Wilson, 1984).
metatarsus, and vertebrae caudalis), and malformations of costae, sternum, and
vertebrae. Statistical analysis was performed using Anova and Duncan's Multiple Table 1. The Average of Fetus Weight, Length, and Hemorrhage
Range Test. Teratogenic effect caused the decrease of fetal weight and length and
the increase of hemorrhage. Ananas comosus extract also inhibited skeletal
ossification (decreased ossification number of metacarpus and metatarsus) and Average
Number of
caused costal malformation (intercostal fusion and convulated costal structure). Treatment
Mother Number of
Weight (grams) Lenght (cm)
Hemorrhage
Keywords : Ananas comosus, teratogenic, skeletal development
Control 5 1,07 ± 0,10 a 24,11 ± 0,58 a 0,00 ± 0,00 a
D1 (20 %) 5 0,54 ± 0,20 b 17,60 ± 3,25 b 5,00 ± 4,64 b
INTRODUCTION
Pineapple (Ananas comosus) contains many nutrients, vitamin A, calcium, D2 (40%) 5 0,56 ± 0,09 b 16,82 ± 1,61 b 7,00 ± 2,24 b
phosphorus, magnesium, iron, sodium, potassium, dextrose, sucrose, and the D3 (80%) 5 0,57 ± 0,19 b 17,04 ± 3,40 b 4,60 ± 4,28 b
enzyme bromelain (a mixture of 95%-cysteine protease) which can hydrolize
proteins (proteolysis) and resistant to heat. Potential of bromelain as painkiller,
antiinflammatory, antiedema, and debridement, accelerate wound healing, and Effect on Fetus Skeletal Development
enhance the absorption of antibiotics, are very useful in postoperative healing All control fetuses had three segment (normal fetus ossification degree) of metacarpus and
(Orsini, 2006). Bromelain is effective as anticoagulant, fibrinolytic, antimetastatic metatarsus. Decreasing number of metacarpus and metatarsus ossification in particularly high-dose
(Maurer, 2001), antileukemia (Baez et al, 2007), anti-cancer, and modulate the because blood levels of enzyme bromelain is high. The molecular weight of this enzyme may less
immune system and hemostatic (Chobotova et al, 2010). Bromelain accelerate than 600 dalton so it can pass the placental barrier. Accumulation of bromelain in the placenta will
menstrual cycle and potential as contraceptive and abortivum. Previous research inhibit transfer of nutrients from the mother to the fetus, then inhibit the metabolism of nutrients
had been carried out on pregnant mice. Extract was given orally 0.2 ml on the day necessary for growth and development of fetal organs, and also minerals needed for calcification.
of gestation 2, 4, and 6 (period of implantation). The extract turned out to be This can disrupt osteogenesis and bone growth.
embryonic lethal (embryotoxic) on the day of gestation 2 and 4 (Mulyoto, 1986). No deformity and abnormal ossification on caudal vertebrae and sternum. No abnormalities were
Teratogenicity is the ability of chemical substance to cause congenital found in the number of costae. Deformities of intercostal fusion ("bridge-like structure") and
abnormalities (structural or anatomical abnormalities) in babies, minor or major convulated costae were only found at highest dose. Costae grow as the development of vertebrae,
malformations and functional disorders. There will be no teratogenicity testing their separation occurs at the beginning of ossification so that disturbances on early development of
done on human. Therefore various methods in animals continue to be developed to vertebrae often followed by abnormal costae (Primmet et al, 1988). Abnormal costae suspected from
anticipate teratology cases. Based on many benefits of pineapple fruit while the the beginning of blastema formation of the vertebrae. Fused costae may be the result of irregularly
negative effect on the implantation period in early pregnancy has been proven, it is direction growth of the vertebrae’s bud and in some places the distance between successive ribs were
necessary to determine birth defects if the mother consumed the fruit during period too close. Adjacent costae, when grows, likely intersect each other, so they will undergo ossification
of organogenesis. together and fused. According to Theiler (1988, in Yantrio et al, 2002), ossification malformations
can be caused by interference of somit, chorda dorsalis, and sclerotom differentiation.
RESEARCH METHODS
Young pineapple fruit was processed to obtain the crude extract (pasta) then Table 2. The Average of Ossification Number of Metacarpus and Metatarsus Segment.
dissolved in distilled water. Twenty pregnant mice were randomly divided into 4
groups. Extract was given orally 0.2 ml/ animal/ day by gavage with 0 (control),
doses 20%, 40%, and 80%. Treatment was given at day 6 to 15 of gestation Number Number of Average of Ossification Number
(organogenesis period). Caesarean section to remove fetus was performed on day Treatment of Observed
18. Fetus morphology observed the fetal length and weight, the completeness of Mother Fetus Metacarpus Metatarsus
front and rear legs, tail, and hemorrhage. Measurement of body length and weight
performed after clearing fetal amniotic fluid from the wrap. Skeleton preparation Control 5 20 3,00 ± 0,00 a 3,00 ± 0,00 a
was done by Alcian Blue-Alizarin Red S staining (Inouye, 1976). Skeletal D1 (20 %) 5 20 1,10 ± 0,55 b 0,70 ± 0,66 b
development observed vertebrae, costae, sternum, metacarpus, and metatarsus. D2 (40%) 5 20 0,65 ± 0,59 c 0,40 ± 0,50 c
Quantitative data were analyzed by Anova and Duncan Multiple Range Test. D3 (80%) 5 20 0,50 ± 0,51 c 0,30 ± 0,47 c

REFERENCES
Báez R, Lopes MT, Salas CE, and Hernández M. 2007. In Vivo Antitumoral
Activity of Stem Pineapple (Ananas comosus) Bromelain, Planta Medica 73(13):
1377-83.
Chobotova K, Vernallis AB, and Majid FA. 2010. Bromelain's Activity and
Potential as An Anti-Cancer Agent : Current Evidence and Perspectives, Cancer
Letters 290(2): 148-56.
Inouye M. 1976. Differential Staining of Cartilago and Bone in Fetal Mouse
Skeleton by Alcian and Alizarin Red S, Congenital Anomalies 161(3): 171-173.
Maurer HR. 2001. Bromelain : Biochemistry, Pharmacology and Medical Use,
Cellular and Molecular Life Sciences 58(9): 1234-45.
Mulyoto. 1986. Kehamilan Mencit yang Diberi Ektrak Nanas Muda (Tesis)
Fakultas Biologi UNSOED, Purwokerto.
Orsini RA; 2006. Bromelain, Plastic and Reconstructive Surgery 118(7): 1640-4.
Price, S.A., and L.M. Wilson. 1984. Patofisiologi, CV EGC, Jakarta, hal.468.
Primmett, D.R.N., C.D. Stern, and R.J. Keynes. 1988. Heat Shock Causes
Repeated Segmental Anomalies in the Chick Embryo, Development 104 : 331-339.
Yantrio, A., Sugiyanto, J., dan Aida Y. 2002. Efek Klorambusil terhadap
Figure 1. Fetus Morphology (ABOVE) : Normal Fetus (left), Hemorrhage (1st and 2nd from the left),
Perkembangan Fetus Tikus Putih (Rattus norvegicus L.) Strain Sprague – Dowley,
Abnormal Fetus/ Dwarf (1st and 2nd from the right);
Jurnal Biota VII(3): 101-108.
Costae (BELOW) : Normal Costae (left), Intercostal Fusion (middle), Convulated Costae (right).
Red colour : cartilage that has undergone ossification, Blue colour : cartilage.
Iriani Setyawati 1, Dwi Ariyani Yulihastuti 2
Laboratory of Animal Structure and Development 1 CONCLUSION
Laboratory of Animal Physiology 2 Young pineapple fruit (Ananas comosus) caused morphological abnormalities, inhibited skeletal
Biology Department, Faculty of Mathematic and Natural Sciences ossification, and caused costal malformation on mouse fetus (Mus musculus L.).
Udayana University, Bali – Indonesia

INTERNATIONAL CONFERENCE ON BIOSCIENCE AND BIOTECHNOLOGY

“PAVE THE WAY TO BETTER LIFE”
UDAYANA UNIVERSITY, DENPASAR – BALI 23-24 SEPTEMBER 2010
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