General Microbiology
General Microbiology
General Microbiology
General microbiology
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MEDIA PREPARATION
Definition:
Microbiological media or bacterial culture media is a medium nutrient that used to prepare for
microbial growth. Bacteria and fungi can grow in microbiological media of various types. This
medium also be used to determine the cause of infectious disease for a specific bacterium.
Microorganism growth is depends on a number of factors such as nutrients, oxygen, moisture
and temperature. The culture medium should be sterile and it should be prevented from the
contamination of a culture with other organisms .This media culture can be classified into two
categories; chemical or organic.
Chemical media uses chemical medium to provide the needed nutrients to an organism. An
organic media contain organic substance that some bacterium may need to grow. This media can
be used to grow variety of bacteria and selective media is used when only a certain type of
bacteria is to be grown under some condition.
Sterilization can be describes as process that destroys or eliminates all forms of microorganisms
including bacteria, viruses, fungi, and protozoa spore and non-spore in order to prevent
contamination of materials and thereby constitute a health hazard. The sterilization process is
depends on the nature of the product, the type of any contamination, and the conditions under
which the final product has been prepared. The Good Manufacturing Practice is required in order
to control all stages of manufacture and sterilization.
METHODS OF STERILIZATION
Moist Heat Sterilization is a technique that exposure the microorganisms to saturated steam
under pressure in an autoclave achieves their destruction by the irreversible denaturation of
enzymes and structural proteins. This sterilization requires precise control of time, temperature,
and pressure to ensure the sterility of the product from any microorganism. The example of
instrument for this sterilization is autoclave. The recommendations for sterilization in an
autoclave are 15 minutes at 121-124 °C. The temperature must be control and monitor the
pressure to obtain the required steam temperature. Figure 1 shows the example of autoclave
Figure 1: Autoclave
Dry Heat Sterilization
Dry Heat Sterilization is a technique of killing bacterial spores and microorganisms using a high
temperature. Dry heat sterilization requires a higher temperature than moist heat and a longer
exposure time. This method is more convenient for heat-stable, non-aqueous materials that
cannot be sterilized by steam because of its deleterious effects or failure to penetrate for example
glassware, powders, oils, and some oil-based injectable. The example of instrument for this
sterilization is hot air oven and microwave. Figure 2 and 3 shows the example of hot air oven
and microwave.
Figure 3: Microwave
Filtration
Filtration is a useful method for sterilizing liquids and gases. Filtration excludes microorganisms
rather than destroying them.It allowing the solution pass through a filter with a pore diameter
that is too small for microbes to pass through. Membrane filtration traps contaminants larger than
the pore size on the surface of the membrane. If contaminants are smaller than the desired
particle, decrease the membrane pore size and trap the product while passing the contaminants
through the membrane. Usually, membranes of not greater than 0.22 μm nominal pore size
should be used. The effectiveness of the filtration method must be validated if larger pore sizes
are employed.
Radiation sterilization relies on ionizing radiation, primarily gamma, X-ray or electron radiation,
to deactivate microorganisms such as bacteria, fungi, viruses and spores. Due to numerous
advantages over heat or chemical based sterilization techniques, this method is particularly
attractive in medicine and healthcare-related fields. Radiation doses should be monitored with
specific dosimeters during the entire process
Types of media
Based on the compositions and the additions of specific components, indicators or chemical,
media can be classified as:-
1. Enrinched media
-Highly nutritious composition containing one or more rich energy source use generally to grow
microorganism in large amount and with ease.
-Helps in promoting the growth of targeted microbe which in normal circumstances may be
difficult to grow
2. Selective media
-Usually enriched and added with specific substance, indicators or chemical such that this media
can now allow only the growth if some selective type of microorganism while not allowing other
microorganism to grow.
-Selective growth media are also used in cell culture to ensure the survival or proliferation of cells with
certain properties, such as antibiotic resistance or the ability to synthesize a certain metabolite. Normally,
the presence of a specific gene or an allele of a gene confers upon the cell the ability to grow in the
selective medium. In such cases, the gene is termed a marker.
-Used to distinguish one microorganism type from another growing on the same media. This
type of media uses the biochemical characteristics of a microorganism growing in the presence
of specific nutrients or indicators (such as neutral red, phenol red, eosin y, or methylene blue)
added to the medium to visibly indicate the de¹ning characteristics of a microorganism. This type
of media is used for the detection and identication of microorganisms. These few examples of
general media types provide some indication only; there are a myriad of di·erent types of media
that can be used to grow and control microbes.
-This type of media uses the biochemical characteristics of a microorganism growing in the
presence of specific nutrients or indicators (such as neutral red, phenol red, eosin y, or methylene
blue) added to the medium to visibly indicate the defining characteristics of a microorganism.
These media are used for the detection of microorganisms and by molecular biologists to detect
recombinant strains of bacteria.
4.
ASEPTIC TECHNIQUES
Aseptic technique is a set of protocols and technique to be observed and followed to safeguard,
minimize and protect the environment and users from exposures to biorisk when working in an
environmentally unsafe environment (biological hazard). These sets of rules are to ensure all bio-
hazardous material remain under strict environmentally controlled environment.
Biological hazard or biohazard defined as a biological substance that can give a threat to the
health of living organism, especially human. It includes a sample of microorganism, virus or
toxic from biological source that can harm human health, animal, plant and other living things.
Working area plays the main role in aseptic technique and need to be sterile to reduce
contamination from airborne particles and aerosols. The working surface needs to be disinfected
thoroughly before and after used. Normal disinfectant used is 70% ethanol.
A working person must have a better knowledge and understanding about personal hygiene
before working with cell culture or other hazardous substance. Good personal hygiene including
washing hands before and after working. A person also must wearing personal protective
equipment to protect themselves from hazardous materials and reduces the probability of
contamination from shed skin as well as dirt and dust from the clothes.
Reagent and media have to be strict quality controlled to ensure their sterility and avoid become
contaminated while handling. Any reagents, media or solution prepared in the laboratory must be
sterile before using the appropriate sterilization procedure; e.g., sterile filter, autoclave.
d) Sterile handling
This is the most important part to ensure the product and user clean from any contamination
occurred while working. There is several cautions need to notes;
Working are and hands need to be sterile with disinfectant; 70% ethanol
The bottles and flasks need to be capped after use and must be sealed with tape
If remove a cap or cover, and have to put it down on the work surface, place the
cap with opening facing down. Better not place the cap on working area.
https://www.aladdin-e.com/up_files/docs/Types%20of%20culture%20media%20used%20in
%20microbiology.pdf
https://www.researchgate.net/publication/315804456_STERILISATION_AND_DISINFECTION
https://bitesizebio.com/853/5-laboratory-sterilisation-methods/
https://www.westlab.com/blog/2018/02/05/different-sterilization-methods-used-in-the-laboratory
https://www.thermofisher.com/my/en/home/life-science/cell-culture/microbiological-culture.html?
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https://www.researchgate.net/publication/332142554_Sterilization_of_Culture_Media_for_Microorgan
isms_Using_a_Microwave_Oven_Instead_of_Autoclave/link/5daa0af3299bf111d4be64d3/download
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