PT BUNGASARI FLOUR MILLS No.

Doc : TP/QC/026
QAQC DEPARTMENT Eff. Date : 01/01/2014
Page : 1/4
SANITATION TEST (SWAB TEST) Rev. No. : 00

1. PURPOSE
Purpose of this procedure is to determine microorganism that still present on sanited tolls and the cleanliness of
workers on production line also to determine the influence of air in production area .

2. SCOPE
This method was applicable for : (1) determine sanitation effectifity on production tools, (2) determine the
frequent of sanitation, (3) determine the source of organism that related to product shelf life, and (4)
determine the contamination of environment by pathogen bacteria.

3. DEFINITION
- Mold is microorganism that did not have chlorofil and have structure as filamen.
- Yeast is one cel microorganism that did not have chlorofil and usually form as miselium (mass of filamen
like yarn).
- Coliform is gram negative bacteria with rod-shaped that usually used as indicator of sanitary quality of food
and water. Bacteria Escherichia coli is group of Coliform, can be found in digestive system of animals and
human, universally present in the feces. The presence of this bacteria is used to indicate fecal
contamination.

4. APPARATUS AND REAGENTS

4.1. APPARATUS

4.1.1. Sterile cotton swab.

4.1.2. Autoclave
4.1.3. Digital balance (0.0001 g).
4.1.4. Vortex.
4.1.5. Pipettor (1ml & 10ml) and sterile tips (1ml).
4.1.6. Pipet Mohr
4.1.7. Sterile petri dish
4.1.8. Screw cap bottle, capacity 250ml and 500ml
4.1.9. Water bath.
4.1.10. Colony Counter
4.1.11. Incubator
4.1.17. Sterile test tube with screw cap
4.1.18. Test tube rack.
4.1.19. Laminair air flow
4.1.20. Ose needle
4.1.21. Measuring glass 100ml
TP/QC/026- Sanitation Test (Swab Test)
 PT BUNGASARI FLOUR MILLS No. Doc : TP/QC/026
QAQC DEPARTMENT Eff. Date : 01/01/2014
Page : 2/4
SANITATION TEST (SWAB TEST) Rev. No. : 00

4.2. SELECTIVE MEDIA and REAGENTS

4.2.1. Selective media for Plate Count Agar, YGC Agar & Chromocult Agar.
4.2.2. Diluent
4.2.3. Alcohol 70%.

5. PROCEDURE

Swab Technique

5.1. Determine the surface of tools or other production object that contact directly with the product.

5.2. Wet the sterile cotton swab with diluent inside the test tube, then squeeze it with pressing the cotton
to upper test tube wall.

5.3. Aseptically, swab the surface that having direct contact with product with widht 5 X 5 cm (25 cm2) using
the wet sterile cotton swab.

Note : After doing swab, the swab area was sterilize with alcohol tissue

5.4. Drop the cotton swab into test tube. To release microorganism from cotton swab, stir the peptone
water on test tube by stirring the cotton swab. Then release the cotton swab from test tube. This
suspension are sample suspension.

5.5. To swab the air, open the petri dish cup that contains selective media for 5 – 10 minutes, depends on
the speed of air on that area. More speed the air, the time needed for swab was shorter.

Inoculation

5.6. Inoculation was doing in laminair air flow. Before it, turn ON the UV lamp to sterilize the area, for about
15-20 minutes (door in close position). OFF the UV lamp, open the door, then turn ON the blower.

5.7. Prepare sterile test tube and petri dish. Attach label on every test tube and petri dish, identify with
number or code of sample, date and dilution factor (10-1, 10-2 etc).

5.8. Using sterile pipet, prepare suspension of sample with dilution factor 10-1, 10-2 etc (as you need).

5.9. Pour the liquid of selective media into each petri dish, with temperature 45  10C as much 13  2 ml. The
selective media used are :

PCA for Aerobic Plate Count

YGCA for Yeast & Mold

Chromocult Agar for total coliform.

TP/QC/026- Sanitation Test (Swab Test)

 PT BUNGASARI FLOUR MILLS No. Doc : TP/QC/026
QAQC DEPARTMENT Eff. Date : 01/01/2014
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SANITATION TEST (SWAB TEST) Rev. No. : 00

5.10. After the media become solid, place the petri dish in upside down position and put it into incubator.
Here are the incubation temperature :

Aerobic Plate Count on temperature 34  10C for 48  2 hours.

Yeast & Mold on temperature 22-250C for 3-5 days.

Total coliform on temperature 34  10C for 24 to 48  2 hours.

5.11. Count every colony on each petri dish that consist 30 to 300 colony.

5.12. Count the colony number based on the widht of surface swab area, number or volume of suspension,
dilution factor and volume of suspension that inoculated into petri dish.

Example :

If there is 80 colony on petri dish that inoculated with 1 ml sample suspension that coming from
dilution of 10 ml swab surface area with widht 25 cm2, then the number of microorganism = 32
cfu/cm2, calculation :

10 1
80 x ------- x ------- = 32 cfu/cm2
1 25

note : cfu = colony forming unit.

5.13. Record the result / number of colony into workbook.

5.14. If there is any deviation, report it to the supervisor immediately.

6. ATTACHMENT

Preparation of Agar and Reagent.

7. REFFERENCE
- FAO, Manual of Food Quality Control 14/12: Quality Assurance in the Food Control Microbiological
laboratory. Rome. 1991.

Approved by, Prepared by,

QAQC Manager QC Assistant Manager

TP/QC/026- Sanitation Test (Swab Test)

 PT BUNGASARI FLOUR MILLS No. Doc : TP/QC/026
QAQC DEPARTMENT Eff. Date : 01/01/2014
Page : 4/4
SANITATION TEST (SWAB TEST) Rev. No. : 00

Attachment
Preparation of Agar and Reagent
1. Plate Count Agar.

22.5g plate count dissolved in 1000ml aquadest and boiled. Placed in screw cap bottle, sterilize on autoklaf
for 15 minutes at temperature 121C. The media should have pH 7.0  0.2

2. Stock Solution.

Dissolved 34g KH2PO4 in 500ml aquadest, adjust pH into 7.2 using NaOH 1N ( 175ml) and dissolved in 1000ml
aquadest.

3. Diluent.

Dissolved 1.25ml stock solution into 1000ml by using aquadest. Sterilize on autoklaf for 20 minutes at
temperature 121C.

TP/QC/026- Sanitation Test (Swab Test)