MICROPARA(LAB)

MEDIA PREPARATION and DISPERSING SIMPLE MEDIA- General Media

Purpose: to grow bacteria
CULTURE,CULTURE MEDIA/COLONY,ASEPTIC
TECHNIQUE ENRICHED MEDIA- Fastidious
Ex: both solid form
SOLID MEDIA -2%-3% agar from seaweeds/culture and  BAP(Blood agar plate )
culture bacteria  CAP(Chocolate Agar Plate)
Ex: DIFFERENTIAL MEDIA-Differential bacteria
 MHA(MUELLER HINTON AGAR) Ex: MAC(McConkey)
 SSA(SALMONELLA SHIGELL)  Pink lactose fermenter
 NA(NUTRITION AGAR)  Yellow montactose fermenter

SEMI-SOLID MEDIA-0.5%-1% Selective Agar –for gram negative

SELECTIVE MEDIA –this is special for mixed cultureisolate
SIM (SULFIDE INDOLE MOTILITY )-most famous pathogenic
semisolid media Ex: SSA(SALMONELLA SHIGELL

LIQUID MEDIA -0 % agar,broth term ENRICHMENT MEDIA- liquid form like broth
Ex:
 Nutrient Broth (NB) CULTURE/COLONY
 BHI (Brain Heart) PURE CULTURE-(1)-Not identified
 TSB(Trypticase Soy Broth) MIX CULTURE-(2 or more)-usually encounter in hospital
normaly get normal flora
SYNTHETIC MEDIA STOCK CULTURE-(1)-identified
Ex:
SEMI SYNTHETIC MEDIA-mixture of natural and synthetic FORMULA:
Dessired Volume:250ml
Dessired volume Quantity =
x Stock Volume:1000ml
LIVING TISSUE –from animal/people Stock volume
Quantity:49.53g
Ex: chic embryo Ex:
250ml
x 49.53g = MAC =12.38/250ml
1000ml
 TUBE
INOCULATION METHOD
Inoculation must de done in aseptic technique 1. LIQUID MEDIA

3 DIFFERENT TYPES OF INOCULATION

Methods for isolation of bacteria
TURBIDITY CLOT
1. STREAK PLATE METHOD –most commonly used
isolation technique
2.SLANTED MEDIA
2. SPREAD PLATE METHOD-quatitative technique
that allows determination of microorganism.

3. POUR PLATE METHOD - quatitative technique that 2.ZIGZAG

allows determination of microorganism.
1.STRAIGHT A LINE
TYPES OF INOCULATING NEEDLE
3.SEMI-SOLID BUTT MEDIA
1. STRAIGHT WIRE-liquid media,semi-
solid,butt&slant
STAB
2. WIRE LOOP –liquid media,slanted,slated media
3/4
3. CALIBRATED WIRE LOOP-urine culture
4.BUTT AND SLANTED MEDIA
4. BENT WIRE-fungal culture
1.STAB 1.STRAIGHT LINE
REMEMBER:
2.STRAIGHT LINE 2.ZIGZAG
*sterilization of wire loop:until red (vertical position)
3.ZIGZAG 3.STAB
*cool for 5-15 less sec.before picking colonies
PLATE D.RADIAL STREAK METHOD

A.INTERRUPTED STREAK METHOD (1 inoculating ½ cm

loop)

90° 90° 1 10° distance

1 1
2 2 2
B.MULTIPLE INTERRUPTED( 4 inoculating loops) E.OVER LAP STREAK METHOD

90° 1 90° 1 90° 1

1 4 90° 1 1
2 3

90°
1 90° 1
1 1

C.MULTIPLE INOCULATION(4 inoculating loop )

Same as Multiple Interrupted but Different bacteria

1
FORMULA:

Dessired volume
x Quantity
Stock volume
Ex:
Dessired Volume:250ml
Stock Volume:1000ml
Quantity:49.53g

250ml =
1000ml

MAC =12.38/250ml