Experiment no.

11
Determination of Thermal Death Point (TDP)
Aim: To determine the Thermal Death Point (TDP) of the given bacterial culture
Principle: The minimum temperature required to kill a bacterial culture when exposed for a
constant period of time is called Thermal Death Point (TDP) for that time. When a series of tubes
containing washed cells of a 24hr culture are exposed to various levels of increasing
temperatures, for a constant time (usually 10mins) and inoculated into a nutrient medium, only
the cells that survived the high temperature exposure will grow to produce turbidity after
incubation at 370C for 24 hrs. The first tube that shows no growth in the series, is the TDP value
for the culture over that time.
Requirements:
1.
2.
3.
4.
5.

Sterile test tubes

Culture suspension
Water bath set different temperatures: at 600C, 700C, 800C, 900C, 1000C
Sterile nutrient broth
Miscellaneous: micropipettes, tips, incubator set at 370C

Procedure:
1. Take a set of 8 sterile test tubes and label them as 600C, 700C, 800C, 900C, 1000C, PC, NC
and MC
2. Dispense 0.1ml of the given culture suspension into each of the first 7 tubes. Do not
dispense any culture in the tube labeled MC.
3. Set the constant temperature water bath to 600C and switch on the equipment. As soon as
the temperature reaches 600C, place the first tube in the water bath for 10 mins.
4. Then reset the water bath temperature to 700C and place the next tube for 10 mins. Repeat
the same for all the tubes, by setting the water bath to its required temperature.
5. Dispense 5ml of sterile nutrient broth to all these tubes except NC (Negative control).
6. The tube labeled PC (Positive control) will have unheated culture and medium. And tube
labeled MC will have only the medium and no culture.
7. Place the tube labeled NC (containing 0.1ml culture) in a boiling water bath at 1000C for
20 mins and dispense 5ml nutrient broth in it.
8. Incubate all the tubes at 370C for 24 hours.
9. Entire procedure has to be carried out in total aseptic conditions.
Observation: After incubation, the tubes were observed for turbidity by shaking gently. It was
seen that NC and MC tubes remained sterile and no turbidity was seen. PC tube showed uniform
turbidity of pure culture. Among the rest of the tubes 600C, 700C, 800C tubes showed growth and
turbidity. 900C and 1000C tubes had no cell growth.

Practical Handout - Microbiology

Sem III (BE Biotech)

Sruthi Pillai
TSEC

Result & Discussion: Thermal Death Point (TDP) of the given bacterial culture was found to be
900C for 10 mins.

Draw this table on LHS:

Table no. 1: observation of cell growth after 24hrs at 370C
Temperature of
exposure for
10mins (0C)

Growth after
24hrs at 370C

600C

700C

800C

900C

1000C

PC

NC

MC

Practical Handout - Microbiology

Sem III (BE Biotech)

Sruthi Pillai
TSEC