• Challenge: write symbol

Starter questions (printed) equation

1. Write down the word equation for:

• Aerobic respiration
• Anaerobic respiration
• Anaerobic respiration in yeast and plant cells
2. Describe the main difference between eukaryotic and prokaryotic
cell structure.
3. What is the function of the flagella and slime capsule?
Title: Growing microorganisms Key words: Binary
fission, culture
LO: Explain how bacteria reproduce and how we medium, agar cell,
study them mutation
Success criteria:
• describe the aseptic technique and explain why its
necessary
• describe how bacteria reproduce by binary fission
• calculate the number of bacteria in a population.

• https://www.youtube.com/watch?v=6-chXVgu8Z0
&ab_channel=SickScience%21
2:09
Why study bacteria?
Are all bacteria harmful?
Bacteria
Useful: food production e.g. milk, yoghurt, cheese, probiotics,
digestion, nitrogen cycle, antibiotic production, nutrient
cycling, protection

Harmful: disease, food poisoning, tooth decay, overgrowth of

harmful bacteria, resistance to antibiotics, pandemics, death
e.g. cholera, bubonic plague
Binary fission Mitosis

Asexual
reproduction 2 identical
daughter cells
For growth
and repair
Simpler Chromosomes
process replicated
Eukaryotic
Cell cytoplasm cells
splits
Prokaryotic
cells
Bacteria divide by binary fission
• Asexual reproduction
• Produces clones.

1. Chromosomes are replicated.

2. Chromosomes line up in the middle and pulled to
opposite ends of cell
3. The cell membrane, cytoplasm and cell wall
separates (cytokinesis)
4. Two identical daughter cells are formed
 Sort them into correct order!

The cell membrane, cytoplasm and cell wall

separates.
Chromosomes line up in the middle and pulled to
opposite ends of cell
Two identical daughter cells are formed
DNA replication - Chromosome set replicated.

Extension: How is asexual reproduction different

to sexual reproduction?
Extension: How is asexual reproduction different to sexual
reproduction?

• That is the main difference between sexual and asexual

reproduction. Sexual reproduction just means combining genetic
material from two parents. Asexual reproduction produces offspring
genetically identical to the one parent.
 Raw chicken can be contaminated with
salmonella and other bacteria
• Under ideal conditions (warmth, nutrients C, N, glucose, salt,
water) bacteria can divide once every 20 minutes.
• You leave chicken out in the morning and go to school. The
room is warm and chicken full of nutrients…
1 salmonella bacterium landed on the chicken.
How many bacteria will there be 8 hours later?
Hint! Convert 8 hours into minutes. Each division
produces 2 new cells.
 How many bacteria grew on the chicken?
The bacteria divides once every 20 minutes
(mean division time)
1. Convert 8 hours into minutes 1. 60 x 8 = 480
2. Divide total time cells have been dividing for minutes
by the time it takes for 1 bacterium to 2. 480 / 20 = 24
produce 2 cells to get the number of 3. 2^24 = 16,777,216
divisions.
(2x2x2x2x2x2x2x2x2x2
3. Multiply 2 by itself for the number of x2x2x2xx2x2x2x2x2x2x
divisions to get the number of cells 2^24 =
2x2x2x2x2x2x2x2x2x2x2x2x2xx2x2x2x2x2x2 2xx2x2 etc.)
x2xx2x2 =16,777,216 cells
 Mean division time
• If we want to estimate the
number of bacteria in a
population we need the
mean division time.

• Mean division time =

average time taken for one
bacterial cell to divide into
two.
 Worked example
A bacterial cell has a mean division time of 30 minutes. How many
cells will it have produced after 2.5 hours?

1. Convert 2.5 hours into mins (2.5 x 60 = 150 minutes)

2. Divide total time they have been producing cells by the mean division
time to get the number of divisions. (150 minutes / 30 minutes = 5
divisions)
3. Multiply 2 by itself for the number of divisions to get the number of
cells 2^5 = 2x2x2x2x2 = 32 cells.

Each cell divides to produce 2 cells, for each round of division the power
increases by 1.
• 2^1 = 2 (1 division)
• 2^2 = 2x2 = 4 (2 divisions) 2 to the power of n
• 2^3 = 2x2x2 = 8 (3 divisions) N = number of divisions
• 2^4 = 2x2x2x2 = 16 (4 divisions)
• 2^5 = 2x2x2x2x2 = 32 (5 divisions)
 Binary Fission Questions
• What two conditions maximise rate of binary fission?
1. A bacterial cell has a mean division rate of 30 minutes. How many cells will
it have produced in 3 hours?
2. A bacterial cell has a mean division time of 24 mins. How many cells will be
produced after 6 hours? Give your answer to 2 significant figures.
3. The mean division time of the bacteria S.aureus is 30 mins. How long will it
take for one of these bacteria to produce a population of 128 cells? Give
your answer in hours.
4. Extension Under different conditions the same bacteria produced 64 cells in
270 minutes. What is the mean division time in these conditions?
 Answers
• 1. 180 minutes / 30 = 6 2^6 = 64 cells
• 2. 6hrs x 60mins = 360 mins
360 / 24 = 15 divisions
2 to the power of 15 = 32 768 cells
to 2 s.f = 33 000 cells
• 3. 128 = 2^7.
So 128 cells is produced after 7 divisions.
7 divisions would take 7 x 30 mins = 210 mins
210 / 60 = 3.5 hours
• 4. 64 = 2 to the power of 6. (divide by 2, 6 times)
the bacterial cell has divided 6 times in 270
minutes
So the mean division time = 270/6 = 45 mins
Bacterial growth
curve
• Bacteria divide every 20
minutes at optimal
conditions (warmth and
nutrients)

• Estimates of cells in culture are

plotted to produce a bacterial
growth curve.
Label the curve – 3 minutes
Checkpoint
• Now that we know how often bacteria grow and the
conditions they need… we can create those conditions
in a laboratory so that we can grow lots of bacteria
colonies and study them!
Sample can be taken using a swab
(coronavirus), Incubating oven –
25C in school labs

Agar plate Nutrient broth solution

Culture
• Method to grow specific bacteria in very large numbers in a lab for
analysis to determine the type of organism and its abundance in
sample
Cells grow in a culture medium (or growth medium) which provides nutrients
for growth
Agar plate Nutrient broth solution
 Bacterial colony
• A bacterium landing on the
agar will divide repeatedly to
form a colony.
• A colony contains millions of
bacteria all identical to one
another
• Each dot is a colony
 3 minutes – link picture to definition
Liquid (nutrient broth) or gel
Culture (agar) that provides nutrients
(sugars, nitrogen) for bacterial
growth.
Culture medium
Method of growing
microorganisms in a lab
Bacterial colony
Bacteria growing on solid media
 Check
point
• We need pure
cultures to study
how effective
antibiotics and
antiseptics are

• How do we make sure only 1 type of bacteria grow in our

culture?
• Aseptic technique
 Sterilise!
sterilise agar (before use)
•  sterilise (Petri) dish before use
• disinfect bench (before use) Autoclave
Use aseptic technique
• pass inoculating loop (through flame)
• secure lid with (adhesive) tape
• minimise exposure of agar / culture
to air / lift and replace lid as quickly
as possible
 The aseptic technique allows for growth of
PURE cultures
• Pure culture is when only 1 type of
bacteria grows on culture medium
• If a culture is contaminated that
means there are non-target
bacteria growing on it
• Uncontaminated cultures of
microorganisms are required to
investigate the action of
disinfectants and antibiotics.
Aseptic technique – growing
uncontaminated cultures of bacteria
Equipment needed:
• Inoculating loop
• Petri dish with agar (agar plate)
• Sticky tape
• Bunsen burner
• Bacteria sample

• Write down the list and as you’re watching the video think about
the method you’d write down.
Aseptic technique
Aseptic technique video
• https://www.youtube.com/watch?v=cuCSELaQ_Go&ab_channel=Davi
dRead
Method prompts
• What is the agar plate for?
• How do you sterilise the inoculating
loop?
• How many times do you repeat it?
• How would you ensure no bacteria
contaminate your sample?
• At what temperature are dishes
inoculated at school and why?
Add to your method
• Prepare agar plate that provides the source of energy for the bacteria
(carbohydrates and nitrogen)
• Sterilise the inoculating loop by placing it in the Bunsen burner flame for a few
seconds (before and after). Also sterilise the culture bottle necks if using liquid.
• Collect bacterial sample and spread the bacteria using a zig-zag motion.
• Repeat three times.
• Tape the lid to make sure no other bacteria enter the agar plate (no
contamination). Make sure air can get in otherwise anaerobic bacteria will
grow.
• In school, Petri dishes are incubated at 25°C to reduce risk of growth
of pathogens that might be harmful to humans. And upside down to prevent
condensation dripping on the bacteria
Testing new antibiotics – required practical
LO: Describe how to investigate the effect of antibiotics on
bacterial growth

What is an antibiotic?
Disc-diffusion
technique
 Effect of antibiotics on bacterial growth
• clean bench with disinfectant solution to kill any organism that could
contaminate our culture
• Sterilise an inoculating loop by passing it through a Bunsen burner flame
• Open a sterile agar plate near Bunsen burner. The flame kills bacteria in the
air
• Use loop to spread bacteria evenly over plate
• Place sterile filter paper discs containing antibiotic onto the pate (different
concentrations)
• incubate at 25C
• Bacteria form a layer.
• Bacteria not growing
= area of inhibition
• Size of area indicated
how effective
antibiotic is
• What is the control?
• Area = pi x r2
(diameter x pi)
(c)     A student tried to grow some bacteria in the laboratory.
The diagram shows some of the apparatus used.

               
This is the method used.
1.   Remove the lid of the Petri dish.
2.   Remove the lid of the bottle containing the bacteria.
3.   Use the inoculating loop to remove some of the bacteria from the bottle.
4.   Spread the bacteria over the agar using the inoculating loop.
5.   Put the lid back on the Petri dish.
6.   Put the Petri dish into an incubator at 25 °C for 24 hours.
Steps 1−5 could cause the sample of the bacteria on the petri dish to be contaminated.
Give three improvements to the method to prevent contamination
•  any three from:
• •   sterilise agar (before use)
• •   sterilise (Petri) dish before use
• •   disinfect bench (before use)
• •   pass inoculating loop (through flame)
• •   secure lid with (adhesive) tape
• •   minimise exposure of agar / culture to air / lift and replace lid as quickly as
possible
• allow:
• •   dip loop into ethanol (after flaming)
• •   keep the lid on the plate for as long as possible
•    or
•    minimise exposure of agar to air
•    or
•    only tilt the lid off (rather than remove it)
• •   flame the neck of the bottle
 Culturing microorganisms questions
1. What is ‘binary fission’? .​
2. Why do you need to sterilise Petri dished and culture mediums before use? 
3. What would you use an inoculating loop for?  ​
4. How do you sterilise an inoculating loop? 
5. How would you secure the lid of the Petri dish? 
6. What temperature would you incubate the samples at in a school and why
should you use this temperature? 
7. How can you test the effectiveness of antibiotics and disinfectants
on bacteria?
8. What is the zone of inhibition? 
 Culturing microorganisms questions
1. What is ‘binary fission’? Cell division where two identical cells to the parent cell are formed.​
2. Why do you need to sterilise Petri dished and culture mediums before use? To kill any
unwanted microorganisms. ​
3. What would you use an inoculating loop for? To transfer bacteria onto the agar.  ​
4. How do you sterilise an inoculating loop? By heating in a Bunsen flame. ​
5. How would you secure the lid of the Petri dish? With tape but not sealed all the way around. ​
6. What temperature would you incubate the samples at in a school and why should you use
this temperature? 25oC, to prevent the growth of pathogens harmful to humans.​
7. How can you test the effectiveness of antibiotics and disinfectants on bacteria? Inoculate
agar with bacteria, place discs soaked in the solutions (water as a control) and place the discs
on the agar containing bacteria. Incubate at 25oC. ​
8. What is the zone of inhibition? An area where bacteria don’t grow. ​
 How long does it take for bacteria If there are 100 bacteria and mean division time is 20, how many will there be in 2
List the conditions that bacteria like to
to divide in good conditions? hours?
grow?

Complete a step by step guide to doing

a culture.

1. List the at least 4 ways to stop contamination

and explain how it works?
1.

2.

2.
3.
Calculate the area of the spore

4.
3.
Challenge
Calculate the area without spore

5.
4.