IN VITRO TESTING OF THE EMBRYO-TOXICITY AND TERATOGENIC

ACTIVITY OF Blumea balsamifera (SAMBONG) LEAF EXTRACT USING

ZEBRAFISH EMBRYO TOXICITY ASSAY

A Research Paper
Presented to the Faculty
of Senior High School
Victoria National High School
Victoria, Tarlac

In Partial Fulfillment
of the Requirements for
Practical Research 2

By:
Sabal, Pearl Angeli M.
Agustin, Trexia Mae E.
Bundoc, Shyneth Marie M.
Cardenas, Jeshel P.
Claveria, Patrick A.
Galicha, Ronel A.
German, Elisha Ann D.
Tagoon, Emere James P.

October 2019
 CONTENTS

Chapter
1 THE PROBLEM AND ITS BACKGROUND…………………. 1
Introduction……………………………………………………… 1
Statement of the Problem………………………………………... 2
Hypothesis……………………………………………………….. 3
Significance of the Study……………………………………….. 4
Scope and Delimitation of the Study………………………….. 4
Definition of Terms…………………………………………….. 5
2 REVIEW OF RELATED LITERATURE AND STUDIES……. 6
Related Literature………………………………………………….. 6
Related Studies……………………………………………………. 8
Local Studies…………………………………………………… 8
Foreign Studies…………………………………………………. 13
Conceptual Framework………………………………………….. 14
3 METHODS OF STUDY AND SOURCES OF DATA…………. 15
Research Design………………………………………………….. 15
Gathering and Plant Identification………………………………. 15
Preparation of the Plant Samples (Sambong)……………………. 15
Hot water extraction of Functional Components…………………….. 15
Preparation of the Treatment……………………………………….. 16
Experimental Treatments…………………………………………. 16
Culture and Maintenance of D. Rerio…. ………………………… 17
Fertilization and Spawning of D. rerio (Zebrafish) ……………… 18
Harvesting and Differentiation of Fertilized Eggs………………… 18
Embryo-Toxicity and Teratogenicity assay……………………… 18
Gathering of Data…………………………………………………. 19
Risk and Safety……………………………………………... …… 20
Statistical Treatment…………………………………………….. 21
4 PRESENTATION, ANALYSIS AND INTERPRETATION OF
DATA……………………………………………………………… 22
Zebrafish Embryo Mortality Rate ………………………………… 22
Zebrafish Embryo Hatchability Rate……………………………… 26
Zebrafish Embryo Delayed Growth/Development…………….. …… 27
Assessment of the Embryo-toxicity and Teratogenic Activity of
Sambong Leaf Extract……………………………………………….. 28
5 SUMMARY, CONCLUSION AND RECOMMENDATI……….. 30
Summary of Findings……………………………………………….. 30
Conclusions………………………………………………………… 30
Recommendations………………………………………………. 30
BIBLIOGRAPHY……………………………………………………………… 23
 ABSTRACT

This study was conducted to specifically assess the Embryo-Toxicity and

Teratogenic Activity of Blumea balsamifera (Sambong) Leaf Extract using Zebrafish
Embryo Toxicity Assay. The embryo-toxicity and teratogenic activity of Sambong Leaf
extract was compared in three different groups namely mortality rate, hatchability rate
and delayed growth of Zebrafish embryo. It was tested with the different concentrations
and each concentration was replicated to avoid uncertainties of data. Cultured Zebrafish
embryos were exposed to different concentrations of Sambong Leaf Extract for until 48
hours. Results showed that the Sambong leaf extract tends to become toxic and contains
teratogenic properties against embryos because as the concentration increases, the mean
percentage of mortality rate and delayed growth of the embryo tends to increase also.
Furthermore, as the concentration increases, the mean percentage of hatchability rate of
the embryos tends to decrease. Hence, the researchers inferred that the Sambong Leaf
Extract still has its embryo-toxicity and teratogenic activity which may cause harm to
embryos. Observing and ensuring the right amount of concentration intake is a very
essential step when using this plant as a medicinal treatment especially to those pregnant
women.