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UNIVERSI1Y Of JORDAN

Faculty OF Medicine
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LECTURE NO: Ll\­


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Microbiology

Lect.NO.4

Date: 21/9/2008

Bacterial growth and nutrition Continued




Bacteria are divided into two major groups:
1- Heterotrophic: mostly pathogens related to our human body.
2- Autotrophic: widely distributed in nature, and rarely associated
with infection.

• The term saprophytic bacteria describes bacteria which


are found in nature, it is generated through food or water
contamination and reach our intestine, but generally with no
significant effect.
.. Culturing bacteria: it is important to isolate bacteria from an
existing specimen like blood, urine, ...
(j Culture growth should be associated with certain growth
conditions like neutral, acid, or alkaline ph. For Ex: we have
a type of bacteria called Vibrio Cholera, the causative agent
of the disease cholera, and this organism despite the fact it
can infect our intestine, it can't be well isolated in a neutral
medium, we have to use alkaline medium with a ph of
8.5209.
• The terms acidophilic, mesophilic, thermophilic bacteria are
related to the temp that supports the optimal growth, which
is important to recognize the organism in clinical specimen.
c Generally the pathogenies are considered the mesophilic
bacteria, which means that they grow in a wide range of
temp between 20 - 40 degrees, where as the acido, and
thermo are less causative agents of diseases.
How can we recognize bacterial growth?
Look at the slide with the title containing MacConKey
• We have two types of media :­
1- Solid medium: where you can recognize on the surface the
presence of some E-coli colonies, this type is called
MacConkey agar, this solid medium should be used to
recognize the colony, No. of colonies, the morphology of
the colony, is it flat or elevated over the surface ... , the color

2- Fluid medium: which we call broth, and it can be used


exactly as a solid medium to grow organisms capable of
living in vitro conditions (out side a living host, as in the
lab), we can take samples from food or water, and cultivate
them, after incubation of 37 degrees which is the optimal
temp for most rapidly growing pathogens, so after 24 hours
we can recognize the presence of particles disturbed in this
medium known as turbidity (which means the presence of
L a particle or bacterial cell in a culture medium broth and
10 min" I then the No. of colonies can be exactly estimated)
@ The bacteria uses for replication a very simple method
called Binary fission, if we have a single cell in a medium
with the appropriate amounts of water and nutrients the cell
will be elongated, the cell produces a septum (cross wall),
**generation time » the time which is necessary for one
single cell to be duplicated. It diffs from one type to the
other, it can be for Ex: in relation to Escherichia coli which
is found in the intestinal tract, and grows under facultative
anaerobic conditions, each generation requires 20 mins, in
24 hr we could have 10/\6 cells, this could be measured by
bacterial growth curve. In our body it well usually try to
suppress such growth but if the bacteria managed to over
come the immune system you can expect in a period of
time between 24-72hr to see what the curve in the slide
represents.
® If we put a cell in a fresh medium (1 L of milk for Ex) for
cultivation, it will be activated, which means all necessary
enzymes will be activated which in turn will induce
metabolic activity, this process varies from one kind to the
other, it might extend from 1-2 hrs, it is called lag phase
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in length).

2-The sec phase called exponential growth phase (the fastest


one), where the No. of cells will witness a logarithmic increase,
within 2-3 we will have one million cells.
3-But in this 1L of milk we oniy have a iimited amount of
nutrients, so within 10hrs it will utilize all nutrients in this
medium and begin to enter the stationary phase, here we
have equilibrium between the new viable cells and others
which begin to die, because we have certain enzymes
between cell wall and cytoplasmic membrane called lysozyme
which trigger the process of cell destruction.
4-The last phase happens after 24hrs all needed oxygen,
nutrients storages are depleted, and this phase happens
slowly it might extend from few days to few weeks.

* In the culture medium the system is closed but in our body it


is opened with a contentious system (highly regenerative, in­
out), so as long as the infected person is alive the bacteria can
utilize more nutrients, and cause damage to all important
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industry, for Ex: if they want to produce a type of organic acid,
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or vitamin, they can do so by using some kind of bacteria and


having a continuous culture medium, supplied by oxygen so
the bacteria will reach the stationary phase and stay there
never reaching the death phase.

Bacteria genetics
It is important to understand the role of antimicrobial drugs in
the treatment of infection, and how bacteria can resist the
various types of antibiotics (with resistance factors), and how
they become toxigenic.

@ Any bacterial cell carries all genetic Into WhiCh are


necessary for growth (production of all necessary enzymes,
and the over all structure). All the structural features are
encoded in the genetic pole of the bacteria (flagella,
capsule, generation time, fimbria ... ), and these genes are
arranged !!1 the chromosomes. The bacterial chromosome
is a single stranded DNA, composed of double helix, each
gene is composed of a small segment of DNA, that is
composed of a small set of nucleotides in the form of triple
codon. The No. of nucleotides can be measured by kilo
base pairs, base pairs means nucleotides ( A -> T, G -> C).
But not all genes are active, some of them are expressed
this is called phenotype expression of the bacterial cell, like
the color of the colony of the culture medium, it also means
the metabolic activity in relation to the utilization of lactose
» if the organism is lactose positive (it can break down
lactose) this is a phenotype. Where as other genes might
be silent, and only under certain conditions they can be
activated and expressed. There for, any type of bacteria
has two properties:

*One called genotype which include all genes in the bacterial


chromosomes.
**And the other phenotype that include all expressed
recognizable characteristics, this can also be recognized in
relation to disease, because certain genes can be in the form
of invasiveness » produce specific enzymes which affect the
mucosa and damage it, or expressed in the production of
toxins, and food poisoning like staphylococcus.

@ The central dogma in relation to replication of bacteria can


be easily demonstrated like follows » the DNA is
replicated to mRNA (transcription), which then is attached
to the ribosome, that translate the massage conveyed to
specific sequences of amino acids (translation), which are
(the amino acids) delivered to the location by tRNA.
liP Bacterial genetics where also introduced in pharmaceutical
industry, by capturing a specific gene from humans, like the
gene of insulin in the pancreas, and transplanting the gene
of insulin into yeast cells, then by allowing them to
replicate, they produce insulin in an industrial scale (this
process is called bioengineering), also this process was
used in producing vaccines for some diseases such as,
10 mins ~ hepatitis A, B, ....
® Another topic of great importance to our subject is H(J)'Ifl:f UIiJ!
diagn({))se diseases?

if we want to identify the causative agent of a disease, such as


pneumonia, we collect a specimen, and then culture it. in the

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sequences in the DNA of the clinical specimen, and due to the


fact that we have certain conservative regions within the
bacterial chromosomes, which are considered as trade marks
for each type, then we can cross mach between the obtained
ones and the previously obtained and identified ones, this
requires a speciai technique known as the peR technique
(Polymerase Chain Reaction).

IF you have a single cell during the infection (this could


happen !! as the immune system greatly reduces the No. of
the infecting cells), it will release a few amount of DNA which
can't be easily characterized without amplification » to
increase the amount of DNA in the clinical specimen
(copies).ln short a very powerful technique, it can identify the
slightest amount of infectious agents in clinical diagnosis.

Ex: we use it in our lab to identify T. B, because the normal T. B


culture requires 5 to 6 weeks to know for sure that the person
is infected with micro bacterial tuberculosis » (an infectious disease
that causes small rounded swellings tubercles to form on mucous membranes, especially a
disease pulmonarv tuberculosis that affects the lungs), where as with PCR it
takes 5 to 6 hrs, so you can take the necessary steps to
isolate the infected person because of the epidemic nature of
this disease.

*This is easier for bacteria than for viruses » sometimes we


have more than one virus that produce the same clinical
picture, like influenza, that is caused by influenza viruses, but
it can also be produced by rino viruses, so it is not easy to
differentiate between the two.

@ As we have said, bacteria contain a single, circular, double


stranded, super coiled DNA, which is considered as a
single chromosome not separated from the cytoplasm by a
nuclear membrane, so it is naked, freely floating within the
cytoplasm, not necessarily concentrated in the center of the
ceii, the iength of this chromosome is 1200 Micrometer, or
roughly 1 Millimeter, so as you can see it is extremely long,
but it is super coiled, and it is considered the genome »
the main genes of the bacteria, and contain (2 - 5) x 10A 6
nucleotides, and this is enough to contain 1000 to 3000
genes, rarely 5000.
II During bacterial replication whither in nature or in the
human body, the DNA strand might lose some nucleotides
or, insert (replace) others » Mutation, this can be of two
types: 1-Under natural condition it is considered as a
spontaneous mutation, and it can't be easily recognized in
culture, like E-coli culture, you can't recognize the diff in
color, because these changes can be very minimal, like
having 10"'7 cells of the same type with 4 to 5 cells with diff
characteristics, but if the diff has increased to
overwhelming levels, that, it causes a change in the
phenotypes of the bacteria, Ex: instead of being susceptible
to penicillin, it develops a certain resistance to penicillin.
2-lnduced mutation, for Ex: by ultraviolet, presence of
chemicals. These two types of mutations can be controlled
by the bacteria to some extent, they contain genes to repair
any damage inflected on the DNA sequence also to some
extent.

*Ex: If we have a bacterial culture of staphylococcus


(staforia ???, the Dr's words) » bacterium occurring in
clusters: a bacterium that typically occurs in clusters resembling grapes, normally inhabits
the skin and mucous membranes, and may cause disease. These bacteria commonly infect
the skin, eyes, and urinary tract, and some produce toxins responsible for septicemia and
food If these bacteria acquired a specific virus
poisoning.
carrying a gene which is responsible for the production of
toxins, the bacteria will produce a strain that produces
toxins contaminating our food, and affecting our health.

THE END

Quotations:
o Knowledge is proud that he has leam'd so much;
Wisdom is humble that he knows no more.
I!ll The most intelligent people are the people of doing without, because
they love what Allah loves and dislike the world which Allah dislikes.
@ Inexperience is what makes a young man do what an older man says is
impossible.

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