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Topic 4 - Part 2 Growth and Reproduction

microbiology notes BIO216

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8 views

Topic 4 - Part 2 Growth and Reproduction

microbiology notes BIO216

Uploaded by

robertnketsang
Copyright
© © All Rights Reserved
Available Formats
Download as PDF, TXT or read online on Scribd
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Microbial Growth and Nutrition

Demonstrate a thorough understanding of:


➢ Bacterial growth and reproduction,

➢ Nutritional and environmental requirements,

➢ Classification of microorganisms based on growth conditions

➢ Factors that affect microbial growth,

➢ Culture media used


Microbial Growth and Nutrition
Learning outcomes:
➢ Describe reasons for (Comprehension): Microbial growth requirements

➢ Analyse (Analysis): the factors that affect bacterial growth

➢ State (Knowledge): Bacterial growth phases

➢ Calculate (Application): bacterial growth rate

➢ Summarise (Comprehension): on the different culture media for bacterial growth

➢ Differentiate between (Analysis): culture media used for bacterial growth

➢ Summarise (Comprehension): the methods used for measuring bacterial growth


Microbial Growth and Nutrition
➢ Environmental factors affecting growth:

➢ Temperature as factor in growth, classes of microorganisms based on


temperatures;

➢ Oxygen as a factor in growth, classes of microorganisms based on oxygen


tension; pH as a factor in growth;

➢ Water activity as factor in growth; Osmotic pressure, osmophiles and compatible


solutes;

➢ Growth and reproduction in bacteria: bacterial growth curve;

➢ growth kinetics: growth rate and generation time;

➢ Batch and continuous cultures;


Factors Affecting Microbial Growth
There are some factors that affect and control the growth of microorganisms
around us, in hospitals, in the laboratory, and in industrial settings.

These factors are:

• Availability of Nutrients

• pH

• Temperature

• Moisture/Osmotic pressure

•Oxygen
Microbial growth requirements:
1. Chemical growth factors/Nutrition:
1-Carbon: organic: Glucose. Inorganic: CO2
2-Hydrogen occurs in organic hydrocarbon molecules and in inorganic molecules
such as water.
3-Oxygen or gaseous requirements of microbes.
4-Nitrogen is used in protein/ amino acid synthesis and nucleic acid polymerization.
5-Phosphorus is essential for nucleic acid synthesis and formation of phospholipids.
6-Sulphur present in certain amino acids such as cysteine and methionine .
Trace elements: K+, Mg2+, Ca2+, Fe2+
➢ cofactors for enzymes

Organic factors: vitamins, amino acids.


➢ (essential organic compound an organism is unable to synthesize)
Acidity (pH requirements):
Pathogenic bacteria grow best at neutral or biological pH which is typically between
pH 6.8 to pH 7.4.

Fungi such as yeasts and moulds prefer slightly more acidic conditions and grow best
between pH 5 to pH 6.

The pathogen Helicobacter pylori is able to survive with pH of the stomach by


producing urease. So it is acidophilic bacterium.
On the other extreme, bacteria that prefer alkaline (basic) conditions are known as
alkaliphiles. Example: Vibrio cholerae (prefers pH 9).
Acidophile Neutrophile Alkaliphiles
Cardinal temperatures:
Temperature requirements: Minimum
Optimum
Psychrophiles: grow below <20oC Maximum
Mesophiles: Grow at optimum between 25oC and 40oC
Normal microbiota and most pathogens grow at or near human body temp of 37oC
Thermophiles: heat-loving microbes of Optima between 40 and 65oC
Hyperthermophile: optimum temperature >80oC
Water activity
➢ Bacteria do not grow on dry surfaces.

➢ In the tropics relative humidity is high; so they can be seen growing on all
kinds of materials.

➢ The ability to withstand dryness has importance for disease transmission e.g.
Mycobacterium tuberculosis.

➢ All bacteria require water for growth.

➢ Water activity has a value of 1.

➢ 100% moisture content = water activity of 1.

➢ A saturated solution of salt has a water activity of 0.8.


Osmotic pressure- draws the water from the cells
➢ most require isotonic solutions
➢ human blood = 0.9% NaCl, isotonic
➢ human skin = ~3-6% NaCl, hypertonic
Most bacteria require 0.9 or above water
activity to grow.
Some are xerotolerant and can grow at 0.6
water activity eg mostly moulds and
yeasts. Those who like very low water
activity are known as xerophiles.
Others like Treponema and Neisseria are
very sensitive and die immediately (0.97).
Water activity lowered by: drying addition of
salt or sugar but there are osmophilic
microorganisms prefer high osmotic
pressure they are referred to as xerophilic
microorganisms as they prefer low water
NaCl concentration
activity
Oxygen tension
1. Aerobes
21% oxygen;

2. Microaerophiles
<21% oxygen, mostly 1-2%;

3. Facultatively anaerobic
Can grow best at 21% oxygen but are also able to grow in its absence though at
a much lower rate.

4. Anaerobes
➢ They grow in the absence of oxygen. They are known as aerotolerant to
differentiate them from obligate anaerobes.

➢ aerotolerants are not affected by oxygen because they do not use molecular
oxygen e.g. Streptococcus, Leuconostoc.
Those for which molecular oxygen is toxic (obligate anaerobes):
There are anaerobes which produce toxic metabolites in the presence of
oxygen. These will die because of the presence of oxygen. e.g Clostridium

Capnophile – require low amounts of O2 but high in CO2. e.g Campylobacter


Why do obligate anaerobes die?

Oxygen exists in several state.

• Superoxide;

• Peroxide

• Singlet oxygen;

• Hydroxyl free radical;

All the above are toxic.


Microbial Growth
Growth

➢ A steady increase in all of the chemical components of an organism;

➢ Growth as increases in number of cells rather than an increase in cell size.

Most bacteria reproduce by asexual process known as binary fission.

Growth of bacteria in lab is used to serve many purposes

Clinical perspective:
➢ Detection, Identification, Antibiotic susceptibility

Industrial objectives:
➢ Biochemical analysis of growth in biomass

➢ To produce desirable products in brewing and


biotechnology industries
The phases of growth
Lag phase Log phase Stationary phase Death phase
• Little to no growth • Period of exponential growth Cell growth = cell death equal to The viable cells
• Period of adaptation; with constant generation cell growth; decline in
• Increase in cell size but time; Phase of unbalanced growth; number
not in number – intense • Very sensitive to physical and Nutrients depleted;
metabolic activity chemical agents; antibiotics Toxic metabolites accumulate;
• Fairly resistant to • DNA replication is faster than Chemical composition of cells
physical and chemical cell division. differ;
agents; Some change from rod to coccus;
Some produce resistant bodies.
Measurement of Microbial growth

Direct methods Indirect methods


Plate counts Turbidity
Filtration Metabolic activity
Direct microscopic count Dry weight
Automated cell count
Methods of quantitating (enumerating) growth:
1. Turbidity measurement
2. Direct microscopic count
3. Viable cell count
Spread plate
Pour plate

All require dilution of samples to obtain manageable numbers


Population growth can be measured in several
ways
1. Turbidity (cloudiness) of a sample can be measured with
a spectrophotometer
chemical analyses
➢ Phosphate
➢ Nucleic acid
➢ Protein
Turbidity (cloudiness) of a
sample can be measured
with a spectrophotometer
chemical analyses
2. Direct Microscopic count
One can perform a direct microscopic
count
Viable cell count
• The pour and spread plate isolation methods allow enumeration of bacterial cells
through dilution of a sample first
The usual practice, which
is most valid statistically,
is to count colonies only
on plates that contain
between 30 and 300
colonies.
Serial Dilution Question
One [1] millilitre sample of river water is added to 99 ml of sterile buffered
peptone water. Ten [10] millilitre of the homogeneous solution is then transferred
into another 90 ml bottle of sterile diluent. Then, 0.1 ml of sample is taken from
the last dilution bottle and consecutively plated on nutrient agar plates in
triplicate.
➢ Calculate the number of bacteria per ml of sample in the original water sample
if the following number of colonies were obtained in the plates after 24 hrs of
incubation 230, 360 and 270.

Colonies counted = 30-300 colonies

(230 + 270)/2 = 250

250 X 100 X 10 = 2 500 000

= 2.5 X 106 CFU/mL

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