Mini ProjectBIO300

Muhammad Solahudin Bin Musa 2011700061Maisarah Binti Mohamad Zulkleple 2011778157Nur Liana Syazwani Binti Zainurin 2011757041Nurul Zahrah Binti Mokhrishah 2011236328

Supervisor : Dr. Noorlis Binti AhmadLecturer : 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. To determine prevalence of Escherichia coli from raw vegetables.

II. To determine susceptibility of antibiotic towards Escherichia coli.

Significance of project

Isolate Escherichia coli

(raw vegetable)

Test effectiveness of several antibiotic

resistance

Create awareness among public

Escherichia coli Taxonomy :

Domain ; Bacteriaphylum ; Proteobacteria

class ; Gammaproteobacteria,order ; Enterobacterialesfamily ; 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.

PreventionTreatmentCause

disease

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 (E.coli)

Sample / Antibiotic clear zone

(mm)

TEC 30

TE 30 KF 30 AK 30 E 15 P 10 C 30Antibio

-grammeSample

LocationPart of Sample

MAR Index

Pegaga13(R)

18(R)

(R)21(S)

12(R)

(R)27(S)

TecTeKfEP SoilLeaves

and roots

0.71

Pucuk ubi9

(R)18(R)

(R)21(S)

17(R)

10(R)

32(S)

TecTeKfEP Food stall Leaves 0.71

Ulam raja (R) (R) (R)21(S)

26(S)

(R) (R) TecTeKfPC Food stallLeaves

and Stem

0.71

Kacang botol

8(R)

16(R)

(R)21(S)

24(S)

17(R)

17(R)

TecTeKfPC Food stall Stem 0.71

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

> 0.2 : high risk source< 0.2 : low risk source

MAR Index calculation:

= 5 = 0.71 7

No of resistanceTotal no antibiotic

used

Figure 3 : Formation of clear zoneafter antibiotic susceptibility test..

DISCUSSIONFactor 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 Surface structure Surface area

Vanderzant, 1992

Bibek Ray and Arun Bhunia, 2008

Doyel, 1989

Dr. Noorlis Ahmad, 2014

Conclusion and Recommendation

12 isolates have been detected as Gram-negative 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