WJEC A2 Unit 3 Biology New Specification 2015 Class Booklet

Unit 3.4: Microbiology

2015 Syllabus content:

Note much of this booklet is covered during your PRACTICAL lessons. We will still go

through the theory as it can be asked as written questions for Unit 3

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 MICROBIOLOGY
MICROBIOLOGY is the study of microscopic organisms and encompasses many subdivisions

including virology, mycology, parasitology and bacteriology.

BACTERIA & FUNGI are important in the RECYCLING OF NUTRIENTS from dead organisms.

Bacteria can RAPIDLY reproduce ASEXUALLY by binary fission (some can reproduce

SEXUALLY)

BACTERIA, FUNGI and viruses all have the potential to cause disease in humans, crops and

domestic animals. BACTERIA can also cause food spoilage

BACTERIA can also be BENEFICIAL to humans (e.g. Gut microflora)

Question (AS revision)

1. Draw a labelled diagram of a bacterium as could be seen under TEM?

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 BACTERIAL CLASSIFICATION
BACTERIA are named according to the BINOMIAL SYSTEM (aka so called binominal nomenclature where
a name is composed of TWO PARTS, both of which (usually) use LATIN GRAMMATICAL FORMS)

e.g. Staphylococcus aureus

= Golden, grape-clustered-berry

Staph = frequently found in the NOSE, RESPIRATORY TRACT, and

on the SKIN

The emergence of ANTIBIOTIC-RESISTANT STRAINS of S. aureus

such as methicillin-resistant S. aureus (MRSA) is a worldwide

problem in clinical medicine.

Escherichia coli (E. Coli) comes from the COLON. Vibrio cholera causes CHOLERA. Bacillus
cereus (BACILLUS is Latin for ROD SHAPED)

All BACTERIA are CLASSIFIED by (i) Shape & (ii) Reaction to Gram stain:

(i) Shape of Bacterial Cells:

The UNIQUE shapes of bacteria are due to the presence of their RIGID (PEPTIDOGLYCAN) CELL WALL.

Three main groups:

- Bacillus
= rod shaped

- Cocci
= round/spherical

- Spirillum
= corkscrew shaped

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 STRUCTURE OF BACTERIAL CELL WALLS
BACTERIA CELL WALL MADE of mixture of polysaccharides & polypeptides called PEPTIDOGLYCAN

or MUREIN.

NAG (N-acetylglucosamine) and NAM (N-

acetylmuramic acid), are two derivatives of GLUCOSE

that form the POLYSACCHARIDE CHAINS THAT MAKE

UP THE BACKBONE OF PEPTIDOGLYCAN in the bacterial

cell wall.
Both have modified 5 carbon rings, with an oxygen beta
bond at the C1,4 bond site.
Note NAG NAM beyond specification

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Cross linking occurs for Cell Wall Strength

Just like cross-linking is critical in

PLANT CELL WALLS with cellulose:

Bacterial cell walls have cross-

linking for strength (protects cell
from bursting (lysis) when water
enters cell PREVENTS OSMOTIC
LYSIS) and to give cell SHAPE.

Drawing - overview of 2x cell wall types:

Key Differences between Cell walls of Gram-negative & Gram positive bacteria:

Gram Negative Gram Positive

PRESENCE of complex outer membrane ABSENCE of complex outer membrane
PRESENCE of LIPOPROTEINS ABSENCE of LIPOPROTEINS
THIN Peptidoglycan wall THICK peptidoglycan wall (relatively)
PRESENCE of LIPOPOLYSACCHARIDES ABSENCE of LIPOPOLYSACCHARIDES

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 GRAM POSITIVE (+VE) BACTERIA
o e.g. Bacillus, Staphylococcus and Streptococcus species.

Gram +ve cells are MORE susceptible to the antibiotic, PENICILLIN* and the antibacterial
enzyme, LYSOZYME (found in our tears from eye).

*PENICILLIN (looked at in unit 4 - Immunology & Disease) is an antibiotic that INHIBITS the
enzyme responsible for the formation of interpeptide linking in peptidoglycan

LYSOZYME (enzyme) catalyses the hydrolysis of 1-4 b linkages between NAG & NAM in
peptidoglycan layers.

GRAM NEGATIVE (VE) BACTERIA

GRAM VE bacteria have chemically COMPLEX walls.
Peptidoglycan is supplemented by large lipopolysaccharides* that protect the cell.
* aka: endotoxins. LIPOPOLYSACCHARIDES are important in IMMUNITY RECOGNITION.
GRAM VE bacteria are LESS affected by LYSOZYME and to PENICILLIN (covered later).

Note in exams this is a quick & easy representation of cell walls.

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Questions

Q2.a) What are the shapes of bacteria:

A _________________________

B _________________________

C _________________________

Q2.b) List the major structural differences between Gram-negative and Gram-positive bacteria?

Gram-negative bacteria Gram-positive bacteria

Q3.a) Lysozyme is an enzyme that is effective against bacterial cells. Explain why?

Q3.b) Penicillin is an effective antibiotic. Which type of bacterial cells is Penicillin most effective
against and what is its mode of action as an antibiotic?

__________________________________________________________________________

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(ii) Bacterial Classification by reaction to the Gram stain

The Danish bacteriologist Hans Christian Gram published the technique in 1884.

THE GRAM STAIN is a means to AID THE IDENTIFICATION of bacteria

Why? The Gram stain utilises the fact that:

GRAM VE and GRAM +VE bacterial cell walls have DIFFERENT staining
properties because of their DIFFERENT cell wall compositions

Note The Gram stain acts on the 3D network of PEPTIDOGLYCAN in bacterial cell walls, therefore
members of ARCHAEA & EUKARYOTES do NOT STAIN because they do NOT contain PEPTIDOGLYCAN.

Key Point 1 the Gram stain is a PROCESS of STAINING & COUNTER STAINING

Key-Point 2
GRAM-POSITIVE bacteria stain Purple because their cell walls RETAIN crystal violet.

GRAM-NEGATIVE bacteria are stained Red by COUNTERSTAIN, as their cell walls

DO NOT RETAIN crystal violet

Magic Word to always used in the Gram Stain descriptions:

= RETAIN

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 THE GRAM-STAIN TECHNIQUE
Mixed bacterial culture Notes
(both Gram ve & Gram +ve)

Bacteria are first FIXED to slide by

quick pass through a Bunsen
burner

Binds to peptidoglycan so ALL

bacteria stain purple / violet

Note - Washing stages are not

shown

Lugols Iodine causes GRAM +VE

cells to RETAIN CRYSTAL VIOLET
(aka Binding fixing)

Washing with ACETONE / ETHANOL

(often mixed) cause crystal
violet be washed from Gram
ve cells (i.e. DO NOT RETAIN
crystal violet)

Example of red dye = Safranin

or Fuchsin = both are known as
COUNTERSTAIN

When cells are microscopically viewed:

Gram +ve are viewed as PURPLE (because they have RETAINED crystal violet stain

Gram ve are viewed as RED (because crystal violet was NOT RETAINED and therefore

stains Red with the COUNTER STAIN.

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 Exam Tip
do not say in exams that Gram-negative bacteria DO NOT stain purple with crystal violet
They both STAIN, but only Gram-positive cells RETAIN the crystal violet.

The Gram reaction reflects the MORE COMPLEX STRUCTURE of Gram-ve cell walls.

Note, you will carry out the gram-stain in the laboratory. You are also supposed to be able to
discuss the theory behind the technique as well for possible Unit 3 questions.

Questions

Q4.a) In the Gram stain procedure; explain the role of acetone-alcohol in distinguishing Gram-negative and
Gram-positive bacterial cells?

Q4.b) why do gram negative bacteria appear red and gram positive bacteria appear violet under the
microscope?

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 CULTURING BACTERIA
Between 1881 1887 Fanny Hesse introduced AGAR JELLY into the laboratory and Julian Petri invented

his (PETRI) DISH both instrumental to culturing microorganisms

CULTURING IS THE WORD WE USE I.E. BASICALLY MEANS GROWING

Defn - CULTURING to maintain (tissue cells, bacteria, etc.) in conditions suitable for growth.

MICROORGANISMS reproduce rapidly in suitable environments.

Rm bacteria divide ASEXUALLY = EXPONENTIALLY (i.e. bacterial population DOUBLES each division)

MICROORGANISMS vary in their optimal growth

conditions of differing NUTRIENTS, PH and/or
TEMPERATURE values.

BACTERIA CAN divide in 20 MINUTES given OPTIMAL

GROWTH CONDITIONS.

FACTORS AFFECTING MICROBIAL GROWTH

MICROORGANISMS require the following FOUR optimised factors for maximal growth rates:
NUTRIENTS, TEMPERATURE, PH & OXYGEN availability.

Note - As well as these FOUR dont ever forget that WATER is essential!!! (and implied) **

Note waste products

NEGATIVELY affect microbial
growth we will discuss these with
full microbial growth curves later

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1. Nutrients

In the LABORATORY, nutrients are supplied in NUTRIENT MEDIA (e.g. NUTRIENT AGAR (SOLID) or
NUTRIENT BROTH (LIQUID)). including:

CARBON usually in the form of GLUCOSE

NITROGEN organic & inorganic form (e.g. Nitrates NO3-). Nitrogen is essential for AMINO ACIDS.
GROWTH FACTORS such as vitamins (e.g. biotin) & mineral salts (Na+, Mg2+, Cl-, SO42-, PO43-)

2. Temperature

ALL cellular growth is ULTIMATELY REGULATED BY ENZYMES.

Therefore, optimal bacterial enzyme range is typically 25-45C.

e.g. optimal temperature
For environmental bacteria is around 25C
for human bacterial pathogens is around 37C (body temperature)

e.g. Enzymes within human pathogens (37oC)

Note - enzymes remain ACTIVE either side of the OPTIMAL TEMPERATURE only the rate of reaction
slows down (until ZERO rate of reaction).

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COOLING DOWN BELOW OPTIMUM slows KINETIC energy so FEWER COLLISIONS

REVERSIBLE (i.e. enzymes return to normal activity if warmed (below optimal)

Note in very cold conditions microbial growth is extremely low due to MEMBRANE
GELLING (i.e. transport mechanisms affected)

HEATING UP BELOW OPTIMUM Causes DENATURING i.e. the BREAKING OF BONDS, both intra-
protein and inter-protein-complexes - disrupting 3D quaternary, tertiary, and/or secondary
structure. this disruption ultimately alters active site / substrate binding.

NON-REVERSIBLE*
When an enzyme is (totally)
DENATURED (i.e. the total
break down of intra-protein
bonds - so Rate of Reaction is
ZERO)

3. pH also significantly affects enzymatic activity

Most BACTERIA and FUNGI have varying pH requirements from slightly ALKALINE conditions to
slightly ACIDIC conditions.
Bacteria tend to favour slightly alkaline conditions (approx. pH 7.4); fungi favour neutral / slightly acidic

Remember that PH OPTIMUM CURVES differ in that deviation of pH either side of the optimum causes
denaturation of the enzymes 3D protein structure (active site) and ultimately LOWER rate of reaction.

i.e. PH DENATURATION is NON-REVERSIBLE*.

Thus BUFFER SYSTEMS in biology are VITAL to maintain enzymatic activity of essential cellular
processes (e.g. amino acids may form Zwitterions).

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 Zwitterions covered in Chemistry unlikely to be in Biology

4. Oxygen availability

Different microorganisms have different REQUIREMENTS for the PRESENCE (or ABSENCE) OF OXYGEN.

OBLIGATE AEROBES
require (i.e. MUST have) OXYGEN

e.g. Mycobacterium tuberculosis* and most fungi (not yeasts)

*Can causes pulmonary tuberculosis

Rm that oxygen is used as the terminal electron acceptor. Such OBLIGATE AEROBES DO NOT HAVE
the pathways to undergo ANAEROBIC pathways

Note details are not required for WJEC

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OBLIGATE ANAEROBES

Can NOT have OXYGEN present

e.g. Clostridium bacteria

(a spore forming bacteria that can cause MOIST GANGRENE
due to production of tissue destroying toxins in a closed
wound).

Note we discuss
CLOSTRIDIUM BOTULINUM
(which produces the toxin: BOTULINUM) in the interaction with THE
SYNAPSE of NERVE CELLS.

Note that BOTULINUM (in low concentrations) can be used for

cosmetic purposes.

This will be looked at later 3.8 the Nervous System

FACULTATIVE ANAEROBES

A facultative anaerobe is an organism that makes ATP (preferentially) by AEROBIC RESPIRATION

HOWEVER - IF OXYGEN is present, facultative anaerobes are ALSO capable of SWITCHING to

fermentation (anaerobic respiration) if oxygen is absent.

e.g. ESCHERICHIA COLI (E. coli) found in

mammalian colon & commonly used in

laboratories (seen later in cloning within Unit 4
Applications of reproduction & genetics)

Nb - We will look at the total bacterial growth curve later in this booklet

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Visual summary of oxygen tolerance in bacteria

Note exam questions are usually based around a diagram similar to the below diagram

Loose fitting cap means oxygen is readily available to surface liquid broth

NOTE - AEROTOLERANT ANAEROBES (beyond specification)

= anaerobes capable of SWITCHING to aerobic respiration

Presence or absence of particular enzymes determines

oxygen tolerance Beyond specification

The presence of enzymes

CATALASE, PEROXIDASE and
SUPEROXIDE DISMUTASE (SOD)

allows TOXIC FORMS OF OXYGEN

to be NEUTRALISED.

AEROBIC & FACULTATIVE

BACTERIA HAVE these enzymes

& therefore TOLERATE OXYGEN

ANAEROBIC BACTERIA do NOT

HAVE these enzymes therefore

-
CANNOT TOLERATE OXYGEN because of the BUILD-UP OF TOXIC PRODUCTS such as O2 (superoxide

anion) OH (hydroxyl radical) and H2O2 (hydrogen peroxide)

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Questions

Q5.a) In addition of mineral nutrients, what are the 2x necessary nutrients required for microbial
growth and give an example of a source of that nutrient?

Q5.b) All enzymes have their optimal temperature range. Describe what happens to the structure of
an enzyme when it is exposed to a greater temperature than its optimum?

Q5.c) What conditions are suggested for preventing the possibility of culturing pathogenic bacteria?

Q6. Escherichia coli is described as a facultative anaerobe. Define facultative anaerobe?

Q6.b) Describe the type of bacteria growing in the following test tubes?

1. ________________________________

2. _________________________________

3. _________________________________

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 DIFFERENT CULTURE MEDIA CAN BE USED
We can CULTURE MICROORGANISMS on DIFFERENT media, broadly defined under 3x categories:

A defined medium

This a MEDIUM which all INGREDIENTS ARE KNOWN

An undefined medium
GFP green fluorescent protein
from jellyfish gene
This is a MEDIUM containing components that ARE NOT ALL
KNOWN.

For example: YEAST EXTRACT and BEEF PEPTONE are both

common sources of amino acids.

A selective medium

A SELECTIVE MEDIUM only ALLOWS only certain bacteria to grow (i.e. SELECTS growth)

For example: MacConkey agar is SELECTIVE FOR Gram-negative bacteria.

ANTIBIOTICS can also be used to

create SELECTIVE media.

For example:
AMPICILLIN can be added to SELECT FOR
bacteria with ampicillin resistance
or
TETRACYCLINE can be added to SELECT

FOR those tetracycline resistant bacteria.

or
PENICILLIN can be added to SELECT FOR
those penicillin resistant bacteria

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 CULTURING BACTERIA & CONTAMINATION
To STUDY BACTERIA, they have to be ISOLATED & CULTURED (i.e. grown repeatedly).

Because MICROORGANISMS are all around us (including the air, work surfaces, our skin, our breath
etc) it is important to PREVENT CONTAMINATION of your growing BACTERIAL CULTURES.

CONTAMINATION = is the unwanted INTRODUCTION of

something other than your desired substance

ASEPTIC TECHNIQUES
= techniques that maintain sterility in apparatus and
prevents contamination of equipment and the
environment by unwanted microorganisms.
Contamination is a TWO-WAY consideration:

I) You do NOT want anything FROM THE ENVIRONMENT getting onto your cultures.

II) Likewise, you do not want your cultures ESCAPING TO THE ENVIRONMENT.

ASEPTIC TECHNIQUES = STERILE TECHNIQUE

ASEPTIC = a procedure performed under STERILE CONDITIONS
STERILE = free from microorganisms

Before culturing bacteria, your equipment & media must be STERILE. STERILISATION can be carried
out in an AUTOCLAVE that is heated under pressure to 121C for 15 minutes.

AUTOCLAVE HEAT STERILISATION

121C 15MIN under Pressure (meaning LIQUIDS do NOT boil over)

Such conditions will kill endospore-producing bacteria (such as
Clostridium).

Endospores are survival structures that are resistant to heat, drying, pH

change & disinfectant.

COMMERCIALLY, Radiation (Gamma rays) can be used to sterilise

plastic equipment because HEAT may damage structure of plastics.

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Good aseptic techniques at the laboratory bench

Once STERILISED, items must continually be PROTECTED to prevent contamination

Work surfaces are STERILISED BEFORE & AFTER experiments using a disinfectant (e.g. 3%

Lysol, or 70% ethanol).

Agar plates always OPENED NEXT TO A BUNSEN BURNER (updrafts eliminate microbes)

The LIDS OF CONTAINERS are NOT PLACED ONTO WORK SURFACES

Lids of PETRI DISHES are opened only JUST WIDE ENOUGH to add samples

NECKS OF BOTTLES ARE FLAMED in the Bunsen burner for 2-3 seconds

INOCULATION LOOPS ARE HEAT STERILISED in the Bunsen burner until glowing hot

Once inoculated, Petri dishes are SECURED WITH ADHESIVE TAPE two pieces only

Agar plates (i.e. made IN Petri dishes) are INCUBATED AT AROUND 25C

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 Note human pathogens usually have optimal growth at 37C under anaerobic conditions
therefore Petri dishes should NOT be completely sealed (cause Anaerobic conditions)
and should be incubated at 25C (prevent optimal growth at 37oC).

DO NOT OPEN PETRI DISHES AFTER INOCULATION (cells have exponentially grown e.g. fungal

spores can be harmful if inhaled).

When finished, cultures must be SAFELY DISPOSED OF (use of AUTOCLAVE)

POPULATION GROWTH CURVES

We must be familiar with BACTERIAL POPULATION GROWTH CURVES and understand EACH stage.

Key Point
BACTERIAL GROWTH CURVES are obtained using a BATCH

CULTURE (closed system) liquid medium, i.e.

NO new nutrients ADDED, and

NO REMOVAL of waste products.

Batch cultures vs Continuous cultures beyond specification

BATCH CULTURES are also used for

industrial PENICILLIN PRODUCTION
(a secondary metabolite)

CONTINUOUS CULTURES are used

for ETHANOL PRODUCTION from

Yeast cultures (ethanol is a primary
metabolite)

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 Drawing Bacterial Growth Curve key stages:

Lag phase

LAG PHASE shows a slow increase in microbial numbers because only a few individuals have been
inoculated initially.
INOCULATE = INTRODUCE (cells or organisms) into a culture medium.

Cells are ADJUSTING TO THEIR NEW ENVIRONMENT in the LAG PHASE where cells become
metabolically active and tend to INCREASE only in CELL SIZE

Exam tip you will NOT get any marks for saying only bacteria are acclimatising / adjusting etc to their new
environment. You must make reference to the following:

Cells begin synthesizing enzymes, DNA & other key

factors needed for CELL DIVISION

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Log phase / Exponential phase / Growth phase

This phase involves exponential* growth (i.e. cell numbers DOUBLE EACH TIME UNIT)
1 > 2 > 4 > 8 > 16 > 32 > 64 > 128 > 256.. etc
Resources are plentiful and conditions OPTIMAL. All cells are adapted (i.e. they have undergone Lag
phase & now have all enzymes and factors for growth and division)
Thus population RAPIDLY INCREASES.

Rate of reproduction >>> (greater than) rate of mortality

Stationary phase

Stationary phase is when the population remains constant, due to a net balance in the numbers of
cells dividing and cells dyeing:

Rate of reproduction = rate of mortality

Do not mention birth rate!!

We can also say that: The carrying capacity* has been reached.

This plateau of the graph is due to such factors as:

a DECLINE in food supply
a BUILD UP of waste products
an INCREASE in competition (in a non-pure culture).

Death / decline stage

Nutrients are EXHAUSTED and/or TOXIC EXCRETORY PRODUCTS ACCUMULATE. Population numbers
decline rapidly until no living microorganisms remain.

Rate of mortality >>> (greater than) rate of reproduction

Note we will cover POPULATION CURVES in the next booklet (4.5 Population size and Ecosystems)

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Questions

Q7. Explain why there is an initial lag in the growth of a microorganism placed into a new culture?

Q8. In terms of population what is happening in the stationary phase of a bacterial growth curve?

Q9. What conditions cause the death phase to occur in a batch culture?

METHODS OF MEASURING BACTERIAL GROWTH

Measuring growth rates of microorganisms is important in ENVIRONMENTAL HEALTH WORK.

For example:
- samples from RESTAURANTS (to check for
pathogenic organisms),
- WATER BOARDS (to check drinking water supplies).

- FOOD MANUFACTURERS ensuring food is fit to eat

Many foods & drugs are produced using microorganisms

grown in LARGE INDUSTRIAL FERMENTERS (200 dm3)

e.g. Penicillin fermentation (from PENICILLIUM fungi) or Ethanol production (from YEAST)
Such processes must be ACCURATELY MONITORED for their population growth

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 TOTAL VS. VIABLE COUNTS
The size of a POPULATION of microorganisms in LIQUID CULTURE can be measured by:

TOTAL COUNTS = quantification of BOTH living & non-living cells.

E.g. using HAEMOCYTOMETER or COLORIMETER (later)

VIABLE COUNTS = quantification of living cells ONLY.

E.g. using SERIAL DILUTION or METHYLENE BLUE stain (dead cells)

SERIAL DILUTION TECHNIQUE

Bacteria can divide exponentially (e.g. E. Coli divides in approx. 20 minutes) under optimal
conditions in a laboratory.

This can lead to bacterial broth cultures containing MILLIONS of cells simply from overnight incubation

We cannot directly count such a broth population, so we need to create a SERIAL DILUTION in order
to be able to have MANAGEABLE NUMBERS of bacteria to count.

We COUNT BACTERIA as COLONIES on nutrient agar plates

A Colony of bacteria = a cluster of cells which arises from a single bacterium by asexual reproduction.
ONE CELL / ONE COLONY THEORY

(A) (B)

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 (A) Shows good SEPARATION of serially diluted individual bacterial colonies. Colonies are easily
counted on such a plate.

(B) Note, only the END of the streak contain INDIVIDUAL separated colonies. There is too much
MERGING of colonies elsewhere for accurate counting.

Colonies must be ISOLATED and NOT grow (MERGE / OVERLAP) into each other.

If a SERIAL DILUTION is insufficient then colonies will MERGE making counting INACCURATE.

If this happens, you should choose an agar plate with a GREATER DILUTION to create ISOLATED colonies.

Setting up of a serial dilution is usually in 10x FOLD steps.

i.e. 1ml of 100 culture into 9ml of media = 10x FOLD dilution = 10-1 dilution.

Drawing Serial Dilution of Broth Cultures:

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To find the total viable cell count the number
of colonies (on the most suitable agar plate
usually between 20-200) are MULTIPLIED
BY THE APPROPRIATE DILUTION FACTOR.

DETERMINING TOTAL COUNTS

When it is desired to count ALL CELLS, living AND non-living. We can use the following techniques,
Haemocytometry and Turbidimetry:

Haemocytometry

Using a haemocytometer is a VERY ACCURATE METHOD. A specialised microscope slide is used to

count BOTH living & non-living cells.

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Turbidimetry a method of measuring growth INDIRECTLY

Using a colorimeter to measure the CLOUDINESS (= TURBIDITY) of a culture i.e.

As cell numbers increase, so will turbidity

A colorimeter CANNOT DISTINGUISH between a living cell or a non-living cell.

Note you are not expected to DESCRIBE the use of a haemocytometer or colorimeter. Just know their
application to non-viable counts

Measuring of diameter of colonies

Rough estimates of growth rates of bacterial or fungal colonies can be made by REGULARLY
MEASURING the DIAMETER OF GROWTH from the centre point on agar plates.

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Questions

Q10. What are the advantages and disadvantages of viable counts vs. total counts?

Q11.a) Why do we carry out serial dilutions for carrying out viable counts of bacteria?

Q11.b) How could you go about measuring a total count for a sample of bacteria?

Q12.a) Which of the three agar Petri dishes would you choose for calculation and why?

Q12.b) Use figures in the above diagram to determine the number of bacteria in the milk sample?

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 Q13

BY4 Jan 2013 WJEC

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 SPECIFIED PRACTICAL EXERCISES
NOTE: Your learning objectives for A2 Biology could contain questions which require you to be able
to UNDERSTAND AND APPLY YOUR KNOWLEDGE of your practical work(s).

Unit 3.4 a) Investigation into the numbers of bacteria in milk

Review Pages 62-63 in your text book AND your experimental lab work.

SCIENCE HUMOUR

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