FOOD MICROBIOLOGY | beafrncsco | 2011033406 | UST-MB

Food Micro
Chapter 4
Detection and enumeration of microbes in food
The Good, The Bad and The Ugly
Good
- Used in the production of food
Bad
- Causes spoilage
Ugly
- Causes human illnesses
APC
- Aerobic plate count
- Estimation of the number of microorganisms in
a food
*Note: rule of thumb is that as the microbial count
increases, the quality of the food decreases\
Sample Collection & Processing
- Representative sample must be collected using
an aseptic technique
- Labeled with
o Name of food
o Date collected
o Other info
- Once the sample is collected it should be
analysed as quickly as possible
o Refrigerated
o Frozen samples should be kept frozen
- if sample is solid, it is mixed with a sterile
diluent
o Butterfields buffered phosphate
o 0.1% peptone water
Processed in a stomacher
Analysis
Drawbacks of surface plating
- Coalescence of colonies
- Growth of spreaders
SPC
- Pour plate
- 1mL is dispensed onto the surface of a petro
plate
- 45C agar is added to the plate
- Disadvatages
o Time
o Expense
o Lack of accuracy
Homogenization
- Separate cells into individual reproductive units
- 1:10 dilutions
In general, 100-mm-diameter plates should
contain bet 25 and 250 colonies
Roll tube method
- Screw cap tubes
- Tube is immersed in cold water

- Incubated upside down

Spiral plater
- Robotic arm to dispense a sample in
Archimedes spiral
- Rotating agar plate
- Decreasing volume of inoculum is deposited
from near the center of the plate to the outside
- Specialized counting grid
- Advantages
o Fewer plates
o Fewer dilution bottles
o Less agar
o Greater number of samples
Membrane filter technique
- Liquid samples that can easily pass through the
filter
- Used to determine microbial numbers in air
Direct epifluorescent filter technique
- Fluorescent dyes to stain bacteria that are then
counted using fluorescent microscope
Hydrophobic grid membrane filter
- Estimate microbial numbers associated with a
variety of food
- 1600 grids
- Mpn is determined
DMC (Direct Microscopic Count)
- Faster than most methods
- No incubation period is required for cells to
metabolize and grow
- Stage micrometer can be used to measure the
diameter
- Simplest and most rapid method
- Changes can be implemented immediately
- Advatantages
o Little processing of samples
o Limited equipment required
- Disadvantages
o Fatigue
o Detection of live vs dead cells
Metabolism-based methods
Metabolism
- Used to determine
o Starch hydrolysis
o Sugar fermentation
o Production of h2S
o Indole
o Nitrate reduction
REDOX
- Energy source : oxidized, another compound :
reduced
- Dyes: (dye reduction test)
o Methylene blue
o Resazurin

FOOD MICROBIOLOGY | beafrncsco | 2011033406 | UST-MB

o Tetrazoliums
Reductase test
- Commonly used in dairy industry
- Completed in a shorter time
Surface testing
- Disadvantage
o Consistent removal of microorganisms
from the test surface
Swab test
- Most widely used method for microbiological
examination
- Moistened cotton or calcium alginate swab
o Addition of sodium hexametaphosphate
- Ideal for rough surfaces
Replicate organism direct area contact
- Direct contact of agar with a test surface and a
use of sticky film or tape
- Raised agar surface
- Not suitable for heavily contaminated or rough
surfaces
Sterile sticky tape
- Commercially available for sampling of surfaces
from equipment to beef carcasses
Sponge system
- Animal carcasses and food contact surfaces
Conventional methods
- Combined with rapid methods to decrease the
time required for identification of
microorganisms found in food