International Journal of Marine Science 2015, Vol.5, No.53: 1-6
2
utilization of bioactive ingredients from algae has not
been done. Based on the biosynthesis process, marine
algae are rich in compounds derived from the
oxidation of fatty acids called oxylipin. Through these
compounds various types of secondary metabolites are
produced (Paul and Fenical, 1983; Paul and Fenical,
1986; Hay and Fenical, 1988; Paul and Puglisi, 2004;
Hay, 1996; Karthikaidevi et al., 2009; Kolsi et al.,
2015).
Halimeda chemically able to produce diterpenoid
metabolites halimedatrial and Halimeda tetra
acetate at
various concentrations. This metabolite has been
observed to play a role in chemical defense against
herbivores, based on their chemical structure and
biological activity. Halimedatrial more effective than
halimedatetraasetat in marine algae defense system to
repel natural enemies (Paul and Fenical, 1983; Paul and
Fenical, 1984; Paul and Fenical, 1986; Paul and Van
Alstyne, 1988; Paul and Puglisi, 2004)
2 Material and Methods
Materials
Seaweed
Halimeda
sp, collected from the waters of
the Gulf of Lampung. Sampling was carried out in
June - July 2014 and analyzed at the Marine Biological
Laboratory Faculty of University of Sriwijaya, Basic
Chemistry and Biotechnology Laboratory LIPI Cibinong.
Samples seaweed washed with running water and
rinsed with sterile water and cut into small pieces.
Subsequently dried and crushed made flour. Halimeda
sp extracted with methanol, evaporated with a rotary
evaporator. Extracts of secondary metabolites identified
by thin-layer chromatography (TLC) or thin-layer
chromatography (TLC). Dry extract sample dissolved
in methanol is used as the test solution, then spotted
by 5 mL of test solution and standard solution on the
plates of silica gel GF 254 as the stationary phase. Put
the plates into the chromatography vessel that has
been saturated with mobile phase consisting of a
mixture of Chloroform-methanol (10: 1) v/v. Elution
until the upper limit of the stationary phase plate.
Identification chromatography with UV light of 254
nm, and then sprayed with cerium sulfate reagent.
Then dried and viewed with UV 254 nm.
Other materials used are pathogenic bacterial culture
types
S.typhi, S. aureus, E. coli
and
B.subtilis
obtained
from laboratory Basic Chemistry and Biotechnology
LIPI Cibinong Bogor. Media for the pace of the
bacteria used are nutrient agar (NA) and liquid
nutrient broth (NB).
The tools used include blenders, autoclaves, incubators,
distillator, pH meter, ose needle, micropipette, magnetic
stirrer, micrometers, shaker, hot plates and oven.
Antimicrobial Materials Selection
At this stage, the analysis of water content materials is
conducted (Apriyantono et al., 1989) and the selection
of materials using solvent extraction of water and
testing activities by the agar diffusion method.
Extraction of materials
The extraction step includes the destruction of
material, the addition of water at a ratio of materials
and water 1:1, 1:2, 1:3 (w/v), then filtering treatment.
The filtrate obtained is sterilized.
b. Testing antimicrobial activity by agar diffusion
method (Wolf and Gibbons, 1996). Nutrient Agar (NA)
which has been sterilized cooled to a temperature of
50o C in a water bath. Each bacterial culture was aged
24 hours at a concentration of 107-108 cells per mlk
inserted into the NA of 40 uL for every 20 ml of NA.
Subsequently made to the cup with a thickness of 4-5 mm.
Then put the paper disc that has been dipped in each
extract Halimeda. Subsequently incubated at 370 C
for 48 hours. Then observed the presence of inhibitory
and in measuring the diameter of inhibition (in mm)
using a micrometer measuring tool. This stage is
carried out with two replications.
3 Result and Discussion
Result
Antibacterial Test Results
Halimeda
sp crude extract was tested by using four (4)
types of pathogen bacteria (
S.typhi, S. aureus, E. coli
,
B.
subtilis
) with treatment four (4) types of
Halimeda
sp
extracts which include:
H.macrophysa, H. incrassata,
H.opuntia
and
H.renschi
. This test is done observation
for 48 hours. In general, the effect of this extract is
significant to the growth of these bacteria (see Table 1).
Phytochemicals Test Results
The phytochemical test results showed extracts
H.renschi
and
H. gracillis
containing steroids and
saponins compounds, while alkaloids, terpenoids,
tannins and flavonoids are not contained in the extract
(Table 2).
