6.1 Microbial Techniques

Edexcel (B) Biology A-level - Microbial Techniques PhysicsAndMathsTutor.
com
Questions
Q1.
Bacteria were cultured in two different types of medium.
One medium contained glucose as the energy source and the other contained sucrose.
The same molar concentrations of glucose and sucrose were used.
Bacteria were grown in each medium for 20 hours.
Every two hours, the numbers of bacteria were determined using dilution plating
and an optical method (turbidity).
The graph shows the results from the dilution plating.
(i) Calculate how many times faster the bacteria in sucrose reproduce than the bacteria in
glucose, during the log phase.
(2)
Answer ...........................................................
Edexcel (B) Biology A-level - Microbial Techniques PhysicsAndMathsTutor.com
(ii) Analyse the data to explain why the growth curve for sucrose is different from the growth
curve for glucose.
(2)
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(iii) On the graph, draw a curve to show the number of bacteria grown in glucose as
determined by the optical method.
(1)
(Total for question = 5 marks)

Q2.
Antibiotics are widely used to treat infections.
Some antibiotics are bactericidal.
Describe how you would use aseptic techniques to transfer bacterial cells growing on an
agar plate to a tube containing a sterile broth.
(5)
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Q3.
Bacteria are the host cells for λ (lambda) phage viruses.
Viruses can be cultured and a growth curve can be produced.
The flow chart shows how this can be done.
The graph shows a growth curve for viruses.

(i) Explain why there was a delay before the number of lysed cells started to increase.
(3)
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(ii) Calculate the mean rate of increase in the actual number of lysed cells between 50
minutes and 90 minutes.
(2)
Answer ........................................................... cells min -1
(iii) A sketch has been made of this growth curve.
Complete this sketch to predict the shape of the growth curve after 120 minutes,
assuming there is an excess of host cells.
(2)

Q4.
Skin flora include bacteria that live on the skin.
Different parts of the body have different types of bacteria present.
Skin flora are beneficial as they inhibit the growth of pathogenic bacteria on the skin.
Skin flora compete for nutrients and space or secrete toxic chemicals.
The following are types of bacteria found in the female genital tract:
• Escherichia coli
• Klebsiella species
• Lactobacillus species
• Staphylococcus species.
(i) Describe how these types of bacteria present in the female genital tract could be
identified.
(4)
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(ii) The photograph shows Lactobacilli on a skin cell from the female genital tract.
Lactobacilli metabolise glycogen into lactic acid.

Explain how Lactobacilli inhibit the growth of pathogenic bacteria in the female genital
tract.
(3)
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Q5.
An investigation was carried out to compare the aerobic respiration of different sugars by
yeast.
A suspension of yeast was used to produce three cultures, each one with a different sugar.
A control was set up that had a suspension of yeast cultured with water only.
The volume of carbon dioxide produced was measured.
The mass of yeast and the concentration of each sugar were controlled.
The graph shows the results of this investigation.
(i) Give one difference between the structure of glucose and the structure of maltose.
(1)
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(ii) Explain the results for the yeast cultured with glucose and the yeast cultured with
maltose.
(2)
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Q6.
An investigation was carried out to compare the aerobic respiration of different sugars by
yeast.
A suspension of yeast was used to produce three cultures, each one with a different sugar.
A control was set up that had a suspension of yeast cultured with water only.
The volume of carbon dioxide produced was measured.
The mass of yeast and the concentration of each sugar were controlled.
The graph shows the results of this investigation.
Explain the results for the yeast cultured with lactose.

(2)
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Q7.
Antibiotics can affect bacteria.
The table describes the mode of action of four antibiotics.
A student compared the effectiveness of these four antibiotics on the growth of bacteria.
Devise a method that the student could use to obtain valid results.
(5)
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Q8.
Food poisoning can be caused by food that is contaminated with pathogenic

microorganisms.
A number of different bacteria can cause food poisoning.
The graph shows the number of new cases of food poisoning after a lunch party.
(i) Explain why it is likely that this food poisoning resulted from food contaminated with
Staphylococcus rather than with Salmonella.
(3)
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(ii) Describe the techniques microbiologists could use to confirm that this food poisoning
was caused by Staphylococcus.
(4)
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Q9.
Salmonella are Gram negative bacteria found in the large intestine of humans.
A scientist studied the growth of Salmonella.
(i) Salmonella was isolated from a mixed culture of bacteria, using streak plating onto
selective media.
Explain why this is a suitable method for isolating the Salmonella.
(4)
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(ii) The scientist made a broth culture of Salmonella at a concentration of 5 × 103 cells per
cm3.
Ten hours later the concentration of Salmonella was 4 × 106 per cm3.
Calculate the exponential growth rate constant (k) for this culture of Salmonella using the
formula
(3)
Answer ...........................................................
(iii) In this calculation, the scientist did not allow for the time that the Salmonella spent in the
lag phase.
Explain the effect that this will have on the calculated value for the growth rate constant.
(3)
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Q10.
Aseptic conditions are used when growing bacteria in laboratories.
Justify two methods, other than UV radiation, for ensuring aseptic conditions.
(2)
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Q11.
Yeast, Saccharomyces cerevisiae, is a single-celled organism used in brewing and baking.
The photograph shows yeast cells seen using a light microscope.
A student counted the number of yeast cells, at 90-minute intervals, growing in a liquid
culture at 30°C.
Describe two precautions that would prevent the yeast culture becoming contaminated with
bacteria.
(2)
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Q12.
culture at 30°C.
The student counted the yeast cells using a haemocytometer.
This is a slide with chambers containing a known volume of liquid. The yeast cells are
counted when viewed using a light microscope.
Any yeast cells totally within the counting square should be counted, along with those which
overlap the top or left-hand side of the square.
(i) The volume of liquid in the counting square shown is 0.004 mm3 .
Calculate the number of yeast cells per mm3.

Give your answer in standard form.
(2)
Answer ........................................................... per mm3

(ii) Identify two causes of inaccuracy in this method of counting cells.

(2)
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Q13.
culture at 30°C.
The graph shows the results.

(i) Calculate the percentage increase in the number of yeast cells from 2 hours to 6 hours.
(2)
Answer ........................................................... %
(ii) Predict the number of cells per cm3 of the culture at 12 hours.
Assume that the culture continues to grow at the same rate.
(1)
Answer ........................................................... × 102 per cm3

Q14.
culture at 30°C.
The student counted the yeast cells using a haemocytometer.
The table shows the number of cells counted using a haemocytometer.
(i) The student calculated the standard deviations using the following formula.
Calculate the standard deviation for the sample at 3 hours.

(3)
(ii) The student concluded that the yeast population doubles every 90 minutes.
Comment on the validity of this conclusion.
(3)
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Mark Scheme
Q1.
Q2.
Q3.
Q4.
Q5.
Q6.
Q7.
Q8.
Q9.
Q10.
Q11.
Q12.
Q13.
Q14.

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Edexcel (B) Biology A-level - Microbial Techniques PhysicsAndMathsTutor.

Bacteria were cultured in two different types of medium.

The same molar concentrations of glucose and sucrose were used.

Bacteria were grown in each medium for 20 hours.

The graph shows the results from the dilution plating.

(Total for question = 5 marks)

Antibiotics are widely used to treat infections.

Some antibiotics are bactericidal.

(Total for question = 5 marks)

Bacteria are the host cells for λ (lambda) phage viruses.

Viruses can be cultured and a growth curve can be produced.

The flow chart shows how this can be done.

The graph shows a growth curve for viruses.

Answer ........................................................... cells min -1

(iii) A sketch has been made of this growth curve.

(Total for question = 7 marks)

Skin flora include bacteria that live on the skin.

Different parts of the body have different types of bacteria present.

Lactobacilli metabolise glycogen into lactic acid.

(Total for question = 7 marks)

The volume of carbon dioxide produced was measured.

The graph shows the results of this investigation.

Explain the results for the yeast cultured with lactose.

Antibiotics can affect bacteria.

The table describes the mode of action of four antibiotics.

Food poisoning can be caused by food that is contaminated with pathogenic

A number of different bacteria can cause food poisoning.

(Total for question = 7 marks)

A scientist studied the growth of Salmonella.

(Total for question = 10 marks)

Aseptic conditions are used when growing bacteria in laboratories.

(Total for question = 2 marks)

Yeast, Saccharomyces cerevisiae, is a single-celled organism used in brewing and baking.

The photograph shows yeast cells seen using a light microscope.

(Total for question = 2 marks)

Yeast, Saccharomyces cerevisiae, is a single-celled organism used in brewing and baking.

The photograph shows yeast cells seen using a light microscope.

Calculate the number of yeast cells per mm3.

Answer ........................................................... per mm3

(ii) Identify two causes of inaccuracy in this method of counting cells.

(Total for question = 4 marks)

Yeast, Saccharomyces cerevisiae, is a single-celled organism used in brewing and baking.

The photograph shows yeast cells seen using a light microscope.

The graph shows the results.

Answer ........................................................... × 102 per cm3

(Total for question = 3 marks)

Yeast, Saccharomyces cerevisiae, is a single-celled organism used in brewing and baking.

The photograph shows yeast cells seen using a light microscope.

The student counted the yeast cells using a haemocytometer.

The table shows the number of cells counted using a haemocytometer.

Calculate the standard deviation for the sample at 3 hours.

(Total for question = 6 marks)

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