INTRODUCTION

1. Test to find bacterial contamination of food (especially

milk).

2. Based on utilization of oxygen by bacteria

which reduces dye and decolorizes it.

3. Faster the decolorization, more inferior is the

bacteriological quality and vice-versa.

Methylene Blue Reductase Test(MBRT)

1. Take 10ml of milk sample in a sterile test tube.

2. Add 1ml of MBRT dye(dye concentration
0.005%).
3. Close the tube with sterilized rubber stopper.
4. Place in stand dipped in serological water
bath maintained at 37±1⁰C.
5. Note time taken for decolorization.
FIGURE: Methylene Blue Dye Reduction
Grading Of Raw Milk Based On MBRT

TIME QUALITY

5 hours and above Very good

3 to 4 hours Good

1 to 2 hours Fair

Less than ½ hours Poor

 Factors Affecting
Ø Rate of reduction depends on the nature of the organism present in

the milk. The rate of reduction by different microorganism is: E. coli

(coliforms) > Streptococcus lactis > faecal Streptococci >

micrococci> Thermoduric organisms > psychrotrophic organisms

Ø Creaming causes bacteria to surface, so the tubes should be

inverted periodically.
 RESAZURIN TEST
• Same principle as MBRT
• More sensitive resazurin dye used

üMix 10 ml milk sample with 1ml of resazurin dye

solution(0.05%).

üPlace stopper and invert the tube to mix.Incubate at

37±1⁰C.

üExamine tubes at the end of an hour in the "one-hour

test" or at the end of three successive hourly intervals in
the "triplereading test."
FIGURE: RESAZURIN TEST
Contd…
Judgment of quality is based either on the color produced
after a stated period of incubation or on the time required
to reduce the dye to a given end-point.

COLOR QUALITY
Blue(no color change) Excellent
Blue to deep mauve Good
Deep mauve to deep pink Fair
Deep pink to whitish pink Poor
White Bad
 ADVANTAGES
• Simple, rapid, inexpensive

• Only viable cells actively reduce the dye

DISADVANTAGES
• Not all organisms reduce the dye equally

• Not applicable to food specimens that contain reducing

enzymes