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Microbes & Disease
Contents
Bacteria
 Culturing Bacteria
 Commercial biotechnology
 Bacterial Disease
 Viral Disease
 Protection Against Disease

Bacteria
A microbe is any organism that is not visible with the naked
eye
 The unaided resolution of the eye is about 0.1mm


Bacteria are classified according to their shape:
1) Cocci: spherical bacteria
- Cocci – smallest bacteria, occur as single spheres
- Diplococci – pairs of spheres, e.g. pneumonia
- Staphylococci – clusters of spheres, e.g. food poisoning
- Streptococci – chains of spheres, e.g. sore throat
2) Bacilli: rod-shaped bacteria:
- Individual rods – e.g. typhoid fever
- Chains of rods – e.g. anthrax
3) Spirilla – large, spiral-shaped bacteria - e.g. syphilis
4) Vibrio – crescent-shaped bacteria - e.g. cholera
Reproduction of Bacteria

Cell elongation results in the synthesis of additional cytoplasm
& nuclear material

DNA replication takes place (there is no mitotic spindle), & the
nuclear material attaches to the plasma membrane or
mesosome

A septum begins to develop, & the nuclear material is
distributed to both sides

The septum is completed, & a cell wall develops to divide the
cell into two

The two daughter cells grow to a critical size, & then repeat
this process
Culturing Bacteria

New genetic material can be inserted into a bacterium in three
main ways:
1) Conjugation: bacteria link together by their pili. Donor
passes a plasmid called the F-factor (fertility) to the recipient
cell. The F-factor may be in a plasmid (replicating
independently), or incorporated into the main bacterial
chromosome
2) Transformation: one bacterium releases DNA which is
absorbed by a second bacterium, allowing it to acquire new
characteristics
3) Transduction: new genes can be inserted into the bacterial
chromosome by a bacteria phage (a virus acting as a vector)
Culturing Bacteria

The bacterial population growth curve occurs in four main
phases:
1) Lag phase: cells are active, but there is little increase in
number. The cells accommodate to the new conditions, take in
water & synthesise ribosomes & enzymes
2) Log phase: Nutrients & space are in plentiful supply, so
there is little competition, & the bacteria multiply at their
maximum rate
Culturing Bacteria
3) Stationary phase: carrying capacity (maximum number of
bacteria that the environment can support) is reached, so
intraspecific competition takes place between bacteria. Hence
the death rate balances the population growth rate, & the
number of bacteria remains roughly constant
4) Death phase: nutrient supply is running out & waste
products accumulate resulting in increased toxicity of
environment. Organisms are killed & population size eventually
falls to zero. Spores may be produced during stationary phase
that are resistant to the adverse conditions
Culturing Bacteria

Bacterial growth can be controlled using physical methods
(gamma irradiation or in an Autoclave using high
temperatures) or by chemical means:
- disinfectants
- antiseptics

Antibiotics can be either:
- narrow-spectrum (affecting a few types of bacterium)
- broad-spectrum (affecting a wide range of bacteria)
Commercial Biotechnology

Microorganisms may be cultured commercially in order to
obtain a substance that they produce

They must be able to:
- Produce the substance in large quantities in a small amount
of time
- Be available in pure culture, & be genetically stable
- Grow rapidly in large-scale culture
- Not be harmful to humans
- Be capable of easy removal from culture
- Grow on readily available & cheap raw materials
- Have growth conditions that do not require extremes of
temperature
Commercial Biotechnology

Yeasts are widely cultured commercially, & are used in the
production of:
- Food substances in their own right, & in bread production
- Beers, ales, lagers & wines, & alcohol as a biofuel

There are two main types of process for the production:
- Batch processes: raw materials & microbes are placed
together in a container vessel. The microbes are then allowed
to grow to their maximum population size, then the fermenter
is emptied & the products are extracted & purified
- Continuous process: the nutrients are continually inputted
into the fermentation vessel & the material is continually
removed & processed. This may be more economical, as it
doesn’t have to be shut down on a regular basis
Commercial Biotechnology

In order to develop a large industrial fermentation process,
four main stages are required:
- Isolation of microorganism: organism must be able to be
purified easily & give the optimum product yield
- Culture preparation: master culture is stored by freeze-drying
(lyophilising) in a small space. Small samples are taken from
master culture in order to produce stock cultures, from which
working cultures can subsequently be produced
- Laboratory scale (200cm3 fermenter): optimum conditions are
determined using a small-scale fermenter
- Pilot plant (200 to 500 dm3 fermenter): laboratory fermenter
is scaled up to make sure that it works on a larger scale. There
may be problems with heating/aeration, which may require
electric mixers & cooling mechanisms
Commercial Biotechnology

Enzymes are produced using submerged culture techniques.
They are produced on a commercial scale when the organism
is in the post-exponential growth phase. In downstream
processing:
- The mixture of nutrient broth, cells and extracellular enzymes
is filtered and centrifuged to remove cellular material
- The liquid enzyme mixture is concentrated by evaporation, to
give the bulk enzyme

The bulk enzyme may be:
- Concentrated by chromatography to give the pure enzyme
- Added to stabilisers to give the bulk liquid enzyme
- Precipitated and filtered, then sprayed and grinded to give
the powdered enzyme
Commercial Biotechnology

Immobilised enzymes are widely used commercially – they can
be immobilised in three ways:
- Cross linkage: the enzymes are linked together by
gluteraldehyde, to form a mesh. This may damage some
enzymes, but those that are not damaged remain very active
- Entrapment: the enzymes are trapped in gel microcapsules or
in a fibrous polymer mesh. This does not damage the enzymes,
but may slow their action due to the substrate having to
diffuse in
- Adsorption: the enzymes are held by weak bonds on the
surface of an adsorbing agent (e.g. glass bead, carbon particle,
collagen). The enzymes easily come into contact with the
substrate, but it is expensive, and the enzymes may become
detached
Bacterial Disease

Most bacteria are either beneficial or harmless to humans –
those that cause disease are pathogens:
- The symptoms of the disease are usually caused by waste
products of the pathogens
- An infection is when the effects are noticeable on the body
- Transmission is when an infection is passed on to somebody
else

Diseases such as typhoid and cholera are transmitted through
water, and can cause diarrhoea

To avoid water contamination, water-treatment processes take
place
Bacterial Disease

Food-borne infections including Salmonella are spread in two
ways:
- By not cooking food thoroughly (e.g. raw eggs: newly laid
eggs may be contaminated with poultry faeces)
- By contaminating cooked meat from handling raw meat first
e.g. chicken

Air-borne infections are spread when an infected person
coughs, sneezes, talks or breathes, as the pathogens are
passed into the air in small droplets saliva, mucus and water

Infections that can be transmitted by direct contact are said to
be contagious

Insect bites can transmit pathogens through the saliva of the
insect
Bacterial Disease

Pathogenicity is the ability of a bacterium to cause disease. The
main factors are:
- The way in which the bacterium attaches and gains entry to
host cells
- The types of toxin produced by the bacterium
- The infectivity of the bacterium (the number needed to cause
an infection)
- The invasiveness of the bacterium (its ability to spread within
the host)

After infection, a pathogen must do three things in order to
produce a disease:
- Attachment
- Entry (penetration)
- Colonisation
Viral Disease

Viruses cannot survive without a living host

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Influenza affects the upper respiratory tract
Acquired immune deficiency syndrome (AIDS) is caused by the
human immunodeficiency virus (HIV), and results in the
immune system breaking down so that it can no longer defend
the body against disease
Viral diseases can be very difficult to treat because the viruses
have no metabolism of their own hence antibiotics have no
effect on them. There are a few antiviral drugs, however, which
can work by:
- Inhibiting the production of viral DNA/RNA by altering the
host cell’s DNA
- Preventing the enzymes essential for the production of new
virus particles from working
- Preventing the viral particles from entering the cells in the
first place
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Protection Against Disease

The human body has a number of barriers that prevent
pathogens from entering the body:
- The skin
- Sebum
- Tears, saliva and urine
- Mucus in respiratory tracts
- Commensals
- Stomach acid and enzymes

If the body is injured, a general inflammatory response will
take place:
- blood clots at the site of a cut, which seals the cut, to prevent
pathogens entering the blood
- inflammation around the site of the injury takes place
- Macrophages are attracted to the site of injury, where they
destroy pathogens by phagocytosis
Protection Against Disease

There are four main types of immunity:
- Active natural immunity: memory cells develop after natural
exposure to antigens
- Active artificially induced immunity: memory cells develop
after vaccination
- Passive natural immunity: antibody transfer (e.g. through
placenta or breast feeding) results in short-term immunity (a
few months), as no memory cells develop
- Passive artificially induced immunity: antibodies are injected
(short-term immunity)

It is difficult to become immune to a virus, as they undergo
many mutations, resulting in many different forms producing
similar symptoms, but with different antigens
Protection Against Disease

Antibiotics are only effective against bacterial infections

Antibiotics work in a number of ways:
- Inhibiting cell wall synthesis
- Binding to ribosomes and inhibiting protein synthesis
- Interfering with prokaryotic DNA replication and transcription
- Binding to the cell membrane to make it more permeable
- Inhibiting cell metabolism

Bacteria may become resistant to antibiotics by altering the
structure of the antibiotic or by modifying the bacterial cells.
Resistance develops as a spontaneous mutation, and can
spread through the population asexually and sexually
Summary
Bacteria
 Culturing Bacteria
 Commercial biotechnology
 Bacterial Disease
 Viral Disease
 Protection Against Disease
