Mini Project

BIO300

Muhammad Solahudin Bin Musa

Maisarah Binti Mohamad Zulkleple
Nur Liana Syazwani Binti Zainurin
Nurul Zahrah Binti Mokhrishah

2011700061
2011778157
2011757041
2011236328

Supervisor : Dr. Noorlis Binti Ahmad

Lecturer
: Madam Izzati Adilah Binti Azmir

ABSTRACT

"Ulam" a traditional raw vegetables, is recognized as a popular main dish in

traditional Malay villagers. It can be eaten simply as it is or typically eaten
with anchovies, cincalok or shrimp paste. Escherichia coli can infects meat
more often than vegetables, but these possibilities do exist for any raw fruits
and vegetables too. In this study, the presence of Escherichia coli was
detected in ulam samples (pucuk ubi, pegaga, kacang botol and ulam raja)
bought from three different outlets in Kuala Pilah, Negeri Sembilan. In order to
clarify the presence of Escherichia coli in four raw vegetables, Gram stain and
KOH (potassium hydroxide) test was done to prove the presence of bacteria
Gram negative Escherichia coli. As for the antibiotic susceptibility testing, two
factors that contribute to the MAR index value are the source of sample and
part of sample. The susceptibility testing of all four "ulam" samples shows the
same value of MAR index of 0.71 which highly resistance towards Teicoplanin,
Tetracyline, Cephalothin, and Penicilin as compare to Amikacin large zone of
inhibition. It is recommended to apply an effective washing of raw vegetables,
maintain cleanliness while handling the vegetables and improve the technique
of cooking raw vegetables in order to kill the Escherichia coli.

INTRODUCTION
Objectives
I.
II.

To determine prevalence of Escherichia

coli from raw vegetables.
To determine susceptibility of antibiotic
towards Escherichia coli.

. Significance
Isolate Escherichia
coli
(raw vegetable)

of project

Test effectiveness
of several antibiotic
resistance

Create awareness
among public

Escherichia coli
Taxonomy :
Domain ; Bacteria
phylum ; Proteobacteria
class ; Gammaproteobacteria,
order ; Enterobacteriales
family ; Enterobacteriacase.
Cell morphology : Bacilli, single.
Gram reaction : Gram-negative
Virulence Factors : Capsule, fimbriae, endotoxin, adhesins ,
exotoxins (enterotoxins), type III secretion system;
strain O157:H7 produces Shiga-like toxin.

Cause
disease

Treatment

Preventio
n

Figure 1 : E. coli structure

Figure 2 : E. coli under compound

microscope (400x of magnification)

METHODOLOGY
Dilution
method

Culturing
method

Isolation
method
Disk
diffusion
method

(Bauer
et al.,
1966)

RESULTS AND DISCUSSION

Sample /
Antibiotic
clear zone
(mm)

TEC
30

TE 30

Pegaga

13
(R)

Pucuk ubi

KF 30

AK 30

E 15

18
(R)

(R)

21
(S)

9
(R)

18
(R)

(R)

Ulam raja

(R)

(R)

Kacang
botol

8
(R)

16
(R)

Sample
Location

Part of
Sample

MAR
Index

TecTeKfEP

Soil

Leaves
and
roots

0.71

32
(S)

TecTeKfEP

Food stall

Leaves

0.71

(R)

(R)

TecTeKfPC

Food stall

Leaves
and
Stem

0.71

17
(R)

17
(R)

TecTeKfPC

Food stall

Stem

0.71

P 10

C 30

12
(R)

(R)

27
(S)

21
(S)

17
(R)

10
(R)

(R)

21
(S)

26
(S)

(R)

21
(S)

24
(S)

Antibio
-gramme

(E.coli)

*Teicoplanin (Tec) Tetracyline (Te) Amikacin (Ak) Cephalothin (Kf) Penicilin (P) Erythromycin
(E) Chloramphenicol (C)

MAR Index calculation:

No of resistance
Total no antibiotic
used
=5
= 0.71
7

Figure 3 : Formation of clear

zone
after antibiotic susceptibility
test..

> 0.2 : high risk source

< 0.2 : low risk source

DISCUSSION
Factor of increase in MAR index:
1. Source of sample

Human (personal hygiene, fecal-oral contamination)

Equipment (dead spot) exp, cutting board & knives

Exposed to the air( not covered)
Flies & vermins
Soil

2.

Parts of sample

Vanderzant,
1992
Bibek Ray and
Arun Bhunia,
2008
Doyel, 1989

Surface structure
Surface area

Dr. Noorlis Ahmad, 2014

Conclusion and
Recommendation
12

isolates have been detected as Gramnegative bacteria (E.coli)

MAR index found to be 0.71 for all the four
samples.
Value that shows more than 0.2 indicates its
from high risk sources.
Recommmendation:
Effective washing of raw vegetables
Maintain cleanliness while handling the
vegetables
Improve the technique of cooking the raw
vegetables

Thank You