NEGROS PRAWN PRODUCERS COOOPERATIVE

ANALYCTICAL AND DIAGNOSTIC LABORATORY

Door No.1 & 2., NOLKFI Bldg., 6th Street., Bacolod City
Tele/Fax 034-4332131
Email Address: nppc_adl@yahoo.com.ph

Client : Rochelle Meagan Espiso Date Submitted: 1/9/20

LSO No: 20-30924 Date Completed: 1/10/20

Specimen: Oregano Ethanolic Extract Control #: 328

Test Conducted: Microbial Assay

Laboratory Test Results:

S. aures Oregano Extract Antimicrobial Assay Result

50% 75% 100% Distilled Water Gentamycin

Disc Zone of Disc Zone of Disc Zone of Disc Zone of Disc Zone of
Inhibitation Inhibitation Inhibitation Inhibitation Inhibitation

(mm) (mm) (mm) (mm) (mm)

R1 10 R1 12 R1 14 R1 0 R1 30

R4 10 R2 12 R2 15 R2 0 R2 30

R3 8 R3 11 R3 14 R3 0 R3 30

R4 10 R4 12 R4 15 R4 0 R4 29

R5 10 R5 11 R5 14 R5 0 R5 31

R6 11 R6 10 R6 16 R6 0 R6 30

R7 11 R7 12 R7 16 R7 0 R7 31

R8 12 R8 12 R8 14 R8 0 R8 30

R9 10 R9 11 R9 15 R9 0 R9 29

R10 11 R10 11 R10 11 R10 0 R10 30

Mean 10 11 14 0 30

Analysis using Kirby-Bauer Disc Diffusion Method:

Method Description:

Preparation of McFarland’s Standard; 100 mL of McFarland’s standard was prepared by mixing

99.5 mL of 1% sulfuric acid and 0.5 mL of 1.175% BaC12.2H2O. The turbidity is equivalent to 1.0 x
108 CFU/mL bacteria. Bacterial Isolates from pure culture of S. aureus were then transferred to sterile
buffered phosphate or distilled water solution and the turbidity was adjusted using a spectrophotometer
at 600nm wavelength in comparison with the absorbance of the McFarland’s standard, 0.089.

Sensitivity Testing for S. aureus. The Sterile swab was dipped it into the broth culture of
organism and was gently squeezed against the inside of the tube in order to remove excess fluid in the
swab. The swab with the test organism was streaked evenly on each of the the sterile and labeled
Mueller-Hinton agar (MHA) plates. After the streaking was completed, the plate to dry for 5 minutes,
then the discs (from different concentration of Oregano Extract) were .placed on the surface of each of
the agar using sterilized forceps or inoculation, a ruler was used to measure the diameter of the zone of
inhibition fob each of the discs used. The measurement obtained from the individual discs were
recorded and compared with the standard table to determine the sensitivity zone to determine whether
the tested bacterial species is sensitive to the different concentrations of Oregano Extract.

NEGROS PRAWN PRODUCERS COOOPERATIVE

ANALYCTICAL AND DIAGNOSTIC LABORATORY
Door No.1 & 2., NOLKFI Bldg., 6th Street., Bacolod City
Tele/Fax 034-4332131
Email Address: nppc_adl@yahoo.com.ph

Study of Bhalodia, N. R, and Shukla, V. J. (2011) revealed that sensitivities of microorganism

species to the plant extracts determined by measuring the sizes of inhibitory zones (including the
diameter of disk ) with values <8 mm were considered as not active against microorganisms.

Reference:
Bhalodia, N. R, and Shukla, V. J. (2011). Antibacterial and anti-fungal activities from leaf extracts of
Cassia fistula 1.: An ethnomedicinal plant. Journal of Advance Pharmaceutical Technology & Research,
2(2), 104-109. http://doi.org/10.4103/2231-4040.82956

NOTE: 1. Result of examination specifically related to samples as received.

2. Test results shall not be reproduced without the approval of the Laboratory Head.

Supervised and Approved for Release:

ROSELYN D. USERO, RCh, MEE

Manager/Laboratory Head

PRC License No. 6460

Form #: 006
 Laboratory Test Results:
T. aures

Oregano Extract Antimicrobial Assay Result

50% 75% 100% Distilled Water Gentamycin

Disc Zone of Disc Zone of Disc Zone of Disc Zone of Disc Zone of
Inhibitation Inhibitation Inhibitation Inhibitation Inhibitation

(mm) (mm) (mm) (mm) (mm)

R1 10 R1 12 R1 14 R1 0 R1 30

R4 10 R2 12 R2 15 R2 0 R2 30

R3 8 R3 11 R3 14 R3 0 R3 30

R4 10 R4 12 R4 15 R4 0 R4 29

R5 10 R5 11 R5 14 R5 0 R5 31

R6 11 R6 10 R6 16 R6 0 R6 30

R7 11 R7 12 R7 16 R7 0 R7 31

R8 12 R8 12 R8 14 R8 0 R8 30

R9 10 R9 11 R9 15 R9 0 R9 29
 R10 11 R10 11 R10 11 R10 0 R10 30

Mean 10 11 14 0 30

Analysis using Kirby-Bauer Disc Diffusion Method: