SLE234 Microbiology Laboratory Report 2 2018

Please answer the questions in the text boxes, short answers and dot points are fine.

1. Give definitions for the following types of bacterial growth in relation to O 2 availability:

Strict Anaerobe:

Facultative anaerobe:

Strict aerobe:

Microaerophilic:

(4 Marks)

2. Describe the bacterial growth that you observed in the cultivation in solid agar in Prac 2

(3 Marks)

E. coli:

Hint, to write 10-110-2 easily etc you can use the following notation 10^-1, 10^-2 etc
Micrococcus luteus:

Clostridium sporogenes:

3. One ml of a sample was added to 99 ml of buffer. One ml of this was plated in nutrient agar.
After incubation, 253 colonies were present on the plate. How many colony-forming units
were present per ml of the original sample? Show your working out.

(2 Marks)

4. You have been given 6 new strains of Bacillus sp. recently isolated from a mine site. In order
to gain some understanding about these strains you set up growth experiments to determine
their growth over 24 hours. A spread plate viable cell count technique was utilised. The
bacteria were grown at 37oC in a nutrient broth medium and samples were taken after 24
hrs. Each sample was serially diluted from 10 -1 to 10-4 and 100 µl aliquots were plated onto
nutrient agar plates to be counted. The plates were incubated overnight and the colony
counts recorded in the table below.
SLE234 Microbiology Laboratory Report 2 2018

Strain number Dilution

10-1 10-2 10-3 10-4

B1 TMTC TMTC 956 110

B2 540 51 5 0
B3 TMTC 480 45 4

B4 TMTC TMTC 670 58

B5 138 13 1 0

B6 908 88 8 0
Note: TMTC = Too Many To Count

Complete the table below by calculating the number of CFU/ml for each of strains listed above (hint:
use the appropriate dilution to calculate CFU/ml)

Strain number CFU/ml

B1

B2
B3

B4
B5

B6
(6 Marks)

5. Devise a scheme to prepare a 10-8 dilution plate, using the least number of dilution tubes. [It
is not practical to pipette any volume less than 0.1 ml and your equipment is similar to that
in the Micro-lab, you only have 10 ml dilution tubes.]

Hint, to write 10-110-2 easily etc you can use the following notation 10^-1, 10^-2 etc

(2 Marks)

6. When using Simmon’s Citrate medium, what indicates a positive reaction? What does this
indicate about the bacterium?
SLE234 Microbiology Laboratory Report 2 2018

(2 Marks)

7. How does the flagella staining technique work?

(2 Marks)

8. How would an old culture of Staphyloccous aureus show up microscopically, after it has been
Gram stained? Why?

(2 Marks)

9. A: In the Kligler test, what change in the media indicates H 2S production?

B: What does a pink coloured colony indicate when using MAC (MacConkey Agar)?

(2 Marks)

A:

B:

(Total= 25 Marks)