Bacterial Quantification by Culture

Question 1: Perfect! You set up a nice little experiment!

What do you need to do next to test if the fungal compound has antibiotic properties?
✓ Incubate the tubes and let the bacteria grow
– Spread the bacteria on an agar plate
– Perform a serial dilution
– Count the bacteria cells

Question 2: How would you interpret these observations?

✓ The fungal compound reduces bacterial growth

– The fungal compound forms complexes with the growth medium
– The fungal compound sterilizes the medium
– The fungal compound sample tube is sterile

Question 3: Let's try an analogy for this quantification. Imagine a farmer just found an old bag of
seeds in his shed. How could you with the least effort figure out how many seeds are still able to
yield a crop?
✓ Only grow a small quantity of seeds
– Slice the seeds open to see if they are rotten
– Seed the whole bag and see how many grow
– Germinate all seeds and count

Question 4: How many living bacteria cells are in one milliliter of the experimental sample? Check
your answer on the computer screen to help.

Page 1
 ✓ 540
– 54
– 5.4 · 104
– 5400

Question 5: Why do you think I stored the sample in the fridge at 4°C?
✓ Growth is slowed down
– To ensure safety of the biohazard material
– Otherwise the sample becomes foul
– E. coli dies at room temperature

Question 6: What is the final volume of bacterial solution in this tube?

✓ 10 mL
– 15 mL
– 9 mL
– 1 mL

Question 7: What is the total dilution factor of this solution from the initial solution now?
✓ 100
– 1000
–1
– 10

Question 8: If you had 1000 cells per mL in your sample. How many would you expect to find in this
0.001 dilution?
✓1
– 100
– 10
– 1000

Question 9: What is the dilution factor of your last dilution?

✓ 104
– -4
– 0.0001
–4

Question 10: While we wait for the incubation, let's think about what we expect. If 1 mL of the initial
solution contained one million cells, how many cells would you expect in 1 mL of the 0.0001
dilution?
✓ 100
–1
– 10,000
– 10

Question 11: Nice! We quantified the bacteria in the two samples. The experimental sample with
fungal compound had less than 1000 viable cells per mL while the control sample had around
2,000,000 viable cells per mL. How can you interpret these results?
✓ The fungal compound slows down bacterial growth
– The fungal compound has no effect on bacterial growth
– The fungal compound is a very potent antibiotic
– The fungal compound kills 99% of the bacteria

Question 12: How did we get from a few thousand cells to more than a million?
✓ Doubling by binary fission

Page 2
 – Very quick rate of meiosis
– Large number of offspring per generation
– Bacteria lay thousands of eggs

Question 13: How do you interpret these results?

✓ The fungal compound slows down bacterial growth
– The fungal compound has no effect on bacterial growth
– The fungal compound kills the bacteria
– The fungal compound is useless as an antibiotic

Page 3