General:

1. Sample analytical unit will change from 25g to 50g which means that the diluent will also change
from 225ml to 450 ml. (increase in PO4 diluent)
For VCO, 50g of sample and 50g of Tween 20 will be added into 400mL PO4 diluent (nasa 1L
po ‘to).
2. The blending time (stomacher) will change from 1 minute to 2 minutes.
3. Tyndallization (3x autoclaving) will perform for sponges and swabsticks. Store at room
temperature after tyndallization.
4. Color coding na rags tapos ito ay sterillized din
a. Green rags- Surfaces (Table tops, pushcarts)
b. Red rags- Windows, Wall, Doors
c. Paper towel- Biosafety and Hand
5. Remind po ulit yung rotation ng disinfectant every 3 months.
6. Hindi na babale ng Isoprophyl alcohol. Ang gagamitin na ay ethyl alcohol 70%. Titimplahin:
a. 175ml ethyl sa 250 distilled water.
b. 700mL na ethyl sa 1000mL distilled water
7. BSA plate should be prepared 24 hrs before use (may Inhibitory property) unused plates after
24 hrs should be discarded.
8. LIA slant must have at least 4 cm butt.
9. Sodium thiosulfate will change from 3ml to 6ml.
10. LAB GOWNS: There must be a separate lab gown for clean room and all around.Should be
change every day to be soaked in 0.5% Lysol which will be replaced every day.
11. Culture media Inventory: “First Expiry out”. Sa remarks ilalagay ang brand ng media.
12. Pouring sa plates( EMBA and BPA) dapat ay 15ml-20ml.
13. Storage of media
a. Unused prepared plates can be stored for 2 to 4 weeks at 5 ± 3 ºC or 1 week at room
temperature, they must be prevented from drying out by sealing petri dish with adhesive
tape along the joint between the lid and the base or by packing several dishes into airtight
plastic bags.
b. Unused prepared culture media can be stored for 3 to 6 months at 5 ± 3 ºC or 1-2 weeks
at room temperature.
14. Hiwalay na timbangan ang gagamitin for sanitizer.

Coliform Count and E. coli Detection

1. WATER ANALYSIS: Kung positive sa LST, may sunod-sunod po sa pagtatanim.

a. EC medium
b. EC-MUG
c. BGBB. (merong daw po kasing inhibitory property si BGBB).
2. 5 colonies with halo ang kukunin sa VRBA for continual analysis.
3. Sa biochem, mula sa PCA slant, magtatanim rin po sa isang LST tube para maconfirmn kung
coliform ito.
4. VP Test (also for Salmonella)
a. Ishake after maglagay ng α-naphtol at VP reagent
b. dapat within 2 hrs lang sya. If negative, lalagyan ng pinch ng creatine (catalyst; para
mapabilis ang reaction)
5. Gram-staining is a MUST.
6. EMBA: 5 colonies ang kukunin w/ or w/o green metallic sheen; as long as merong black
centered colonies, itatanim sa PCA for Biochem
7. PETRIFILM: kung TNTC, magcount ng isang box (estimate), then multiply by 30.

Salmonella:
 1. May inventory po tayo sa VIDAS. Gagawan na rin po ang MDS para maidentify yung frequency
ng paggamit natin nung kit.
2. Once na nagpositive sa VIDAS, rerun the sample (from the same pre-enrichment) using VIDAS
(ulit) at 3M MDS. Then magtransfer na rin sa RV at Tetra for conventional.
3. Any positive results from rapid test ay magbabase sa result ng presumptive test ng conventional
(TSI at LIA)
4. LDB test: Bromcresol purple dye (pH indicator) ang gagamitin kung alanganin ang color result.
Hindi na po mineral oil.
5. Serological test ay gagawin kasabay ng biochem. (Coordinate with jasper how to perform)
6. For BSA plates

Incubation Period TYPICAL ATYPICAL

If present If absent
AFTER 24HRS Get 2 colonies Get 2 colonies Do not pick
any colony
AFTER 48 HRS Get additional 2 Get additional 2 Get 2 colonies
colonies colonies
7. For TSI and LIA:

TSI
K/A (w/ or w/o blackening)
K/A (w/ or w/o blackening)
K/K (w/ or w/o blackening)
A/A (w/ or w/o blackening)
LISA
K/K (w/ or w/o blackening) TSI and LIA biochem
K/A (w/ or w/o blackening) TSI biochem
K/K (w/ or w/o blackening) LIA biochem
K/K (w/ or w/o blackening) LIA biochem

Water Sampling:

1. 200ml of water should be drawn then if it is chlorinated directly put it in the sampling bottle with
6ml sodium thiosulfate.
2. Once na may high counts, report kaagad. Then resample.
3. Makikiincubate po tayo sa 45.5°C ng DCN ng ating EC at EC MUG.

Swabbing:

1. 5mL D/E broth na po ang gagamitin as diluent instead of PO4. Direkta na rin itong dadalhin sa
loob ng production. (magdala ng extra paper na susulatan ng names ng personnel/ equipment).
2. For equipment, laundered at packaging swab naman, 5 areas per equipment na tig 50 sq. cm.
gamit ang isang moistened swabstick. Kakanawin sa D/E ang swabstick at tatanggalin ang
excess diluent mula sa swabstick bago gamitin/magswab sa panibagong area.
3. For personnel, separate na ang swab for left and right hand. Meaning, dalawa na ang sample
sa kada tao. (ex: 1. Capili (left), 2. Capili (right) )
4. Yung mga historically high counts na equipment ay magplaplate ng 1mL at 0.1mL.

Staphyloccocus aureus:

1. Uubusin lang ang stock ng BPA tapos magshishift na sa pure petrifilm.