Keywords: Anthelmintic activity, Methanolic Extract, Myrtaceae, antimicrobial, Syzygium malaccense
Syzygium malaccense (L.)Merr. & Perry is a small tree indigenous to regions of Malaysia and South East Asia . It
is mainly found in lowland to mountain rain forest , .
The tree is valued for its large edible fruits. The fruit of
used in traditional medicines for mouth infections, throat infections, stomach ache and abdominal ailments , .
Crushed leaves are used as antiemetic, purgative and also to treat bronchitis, tongue inflammation, dysentery,
on anthelmintic and anti microbial property, the present study is conducted to evaluate these properties.
The leaves of the plant Syzygium malaccense (L.)Merr. &Perry (Family: Myrtaceae) was collected from Ernakulum
district, Kerala, India in September 2014. The fresh leaves were taxonomically identified by Botanist. The
authentification of the plant was done by Mrs.Leela.M, Botany Department, Paliyam Government Higher Secondary
School, Paravur , Cochin, Kerala, India.
The leaves were dried under the shade and then powdered (40 mesh size) with a mechanical grinder and stored in air
tight container. The dried powdered material of the leaves was then extracted with methanol in soxhlet apparatus.
The extract is evaporated by using a rotary evaporator.
All the Chemicals and reagents were procured from (S.D. Fine chemicals, Mumbai, India). The solvents used for the
study were of analytical grade. The drug Piperazine citrate was procured from Glaxo smithkline Ltd., Mumbai.
The antimicrobial activity was carried out by Kirby Bauer disk diffusion method i.e. Disc Diffusion Method 
are Proteus mirabilis (gram negative bacteria) and Candida albicans .YPD plates were prepared and allowed to
solidify for 5 min and 0.1% of inoculums suspension was swabbed uniformly and the inoculums were allowed to
dry for 5 min. The different concentrations of extracts (1, 2, and 4 mg/disc) were loaded on 5mm sterile individual
discs. The loaded discs were placed on the surface of the medium and the compound was allowed to diffuse for
5min and plate was kept for incubation at 37
̊C for 24 hours. Negative control was prepared by using respective
plant extract and Ciprofloxacin and Clotrimazole were used as positive control for Proteus and Candida albicans
respectively. After 24 hours of incubation, inhibition zones formed around the disc evidence the antimicrobial
activity. Diameters of the zone of inhibition were measured with transparent ruler in millimeter.
The anthelmintic activity was carried as per method of Ajaiyeoba et al method , .
The assay was performed in-
intestinal round worm parasites of human beings. Earthworm Pheretima posthuma (pheretimidae) have been widely
used for the initial evaluation of anthelmintic compounds in vitro, because of their easy availability. Indian adult
earth worm (Pheretima posthuma) was collected from the water logged areas of soils and washed with normal saline
to remove all filthy matter. The earth worm of 3-5 cm in length and 0.1- 0.2 cm in width were used for all the
experimental protocol. The collected worm was authenticated by Anitha P.B, Department of Zoology, GHSS
Test samples and standard solution were prepared by dissolving or suspending 3g of each extract in 30 ml of
and 50mg/ml were prepared from above stock solution. Standard used in the assay is Piperazine citrate 10mg/ml. Six
adult earth worms were placed in petridish which contains different concentrations 10,20,30,40, and 50 mg/ml. The
time taken for paralysis and death was noted. The mean of paralysis time was observed and noted .The observations
were made for time taken for the paralysis, when no movement of any sort could be shown except when the worm
were shaken vigorously. Time for death of the worms were recorded after ascertaining that worm neither moved
when shaken vigorously nor when dipped in warm water(50
The study has shown that the leaf extract is effective against both Candida albicans & Proteus bacteria. The zone of
inhibition of methanolic leaf extract of Syzygium malaccense (L.) Merr .&Perry for Candida albicans ranged from
0.4mm to 0.9mm and for Proteus bacteria, ranged from 0.5mm to 0.9 mm and showed a moderate degree of
antimicrobial activity. Sensitivity of bacterial strains to standard antibiotic (Ciprofloxacin) showed a zone of
inhibition of 1.4 mm and sensitivity of fungal strains to standard (Clotrimazole) showed a zone of inhibition of 1.2
mm which is depicted in Figure 1 and 2 .The results are shown in Table 1 and 3. Minimum Inhibitory Concentration
(MIC) is the lowest concentration at which no growth of test strain was observed. The MIC of methanolic extracts
of Syzygium malaccense (L.)Merr.&Perry for Candida albicans was found to be 10µg/ml and are shown in Table 2
and 4. The study also reveals a concentration dependent anthelmintic activity. The methanolic extracts of Syzygium
min ] at 10mg/ml concentration compared with standard Piperazine citrate [(P) of 25.87 min] at same concentration.
The results are shown in Table 5. Piperazine citrate by increasing chloride ion conductance of worm muscle
membrane produces hyper polarization and reduced excitability that MIC of the methanolic extract for Proteus
bacteria was found to be 10µ g/ml. The results leads to muscle relaxation and flaccid paralysis. So, the methanolic
extract of Syzygium malaccense (L.)Merr. &Perry showed significant antimicrobial activity and anthelmintic
Table 3: Antifungal Activity by Zone of Inhibition
Sample taken (mg)
Zone of inhibition of
Zone of inhibition of
Table 5: Anthelmintic activity of methanolic leaf extract
Time taken for
paralysis & Death
Control (Distilled Water)
Piperazine Citrate (Standard)
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