Aseptic Technique

Aseptic Techniques
• Aseptic technique is a procedure that is performed under sterile

conditions, a method that prevents the introduction of unwanted
organisms or contaminants into an environment.
• A fomite is any inanimate object or substance capable of

transporting pathogens from one medium or individual to
another.
• Aseptic technique is essential in the microbiology laboratory to

prevent any contamination of laboratory personnel, cultures,
supplies, and equipment.
• Flaming is one of the physical methods of sterilization and must

always be applied to the lips of test tubes and also of flasks
whenever culture liquid is poured from one container to another.
Sterilization
• It’s the complete removal of all other microbial forms, including viruses, bacteria, fungus,
spores, and other vegetative cells from the surface or the culture media.
• Based on the purpose of the sterilization, the method is categorized into two groups:
ü Physical Methods: It involves the killing of contaminants or microbial forms using heat,
sunlight, drying, filtration, or irradiation techniques (e.g., UV, infrared, gamma radiation,
and X-ray).
ü Chemical Methods: It utilizes chemicals such as phenol (and any other related
compounds), dyes, soaps, detergents, alcohol, gaseous compounds, and heavy metals
and their compounds to destroy microorganisms.
Disinfection
• Disinfectants are applied to inanimate surfaces, medical equipment, and other man-made
objects whereas antiseptics are used to disinfect skin.
• The term disinfection refers to the use of a physical process or the use of a chemical agent
to destroy vegetative microbes and viruses.
• This does not include bacterial endospores.
• Substances that kill bacteria are bactericidal and those that interfere with cell growth and
reproduction are bacteriostatic.
• Disinfectants and antiseptics are bactericidal and bacteriostatic depending on the
concentration applied.
• The type of disinfectant to be used depends on the surface or material to be disinfected.
Sanitization
• Sanitization is applied in everyday life in order to control possible infections or spoilage of

substances those do not require sterilization, disinfection, or antisepsis but need to reduce
microorganisms.
• Often the sanitizer used is a compound such as soap or detergent.
• Degermation is the process by which the numbers of microbes on the human skin are
reduced by scrubbing, immersion in chemicals, or both.
Culturing Techniques
• Microbiologists use five (the five “I’s”) basic procedures to examine and characterize
microbes:
ü Inoculation
ü Incubation
ü Isolation
ü Inspection (observation), and
ü Identification
Inoculation
To culture a microorganism a small sample, the inoculum, is introduced into a culture medium
usually with a platinum wire probe streaked across its surface. This process is
called inoculation and the growth that appears on or in the medium is the culture. A culture can
be pure—containing one type of organism, or mixed—containing two or more species.
Isolation
Isolation is a microbiological technique in which a specific microbial strain is isolated from a

mixed culture of microorganisms by culturing the microbes on a selective culture media.[3]
However, the procedure must be repeated several times to eliminate contamination by other
microbes and achieve a pure culture of the microbial strain, which is then observed in culture
plates as discrete/isolated individual colonies.
Culturing techniques
Microbes are grown in labs on culture media, which supply their nutritional requirements. These
requirements vary for different microorganisms, thus a spectrum of culture medium recipes have
been developed by scientists to obtain the desired microbial strain.
•Simple or basal media: It consists of sodium chloride, peptone, meat extracts, and water, for
example, Nutrient Broth.
•Complex media: This contains an additional special ingredient that helps to enhance a special
characteristic or provide nutrients for the growth of certain microbes. It may contain extracts
from plants, animals, and yeast, such as blood, yeast extracts, serum, milk, meat extracts,
soybean digests, and peptone.
•Synthetic or defined media: It’s used for research purposes. They are prepared by following
an exact formula and mixing distilled water with specific amounts of inorganic and organic
chemicals.
• Special media: The basic medium supports the growth of a broad spectrum of microbial
forms. However, a special growth condition is required for the culture and isolation of only
a certain type or selected strain of bacteria. These formulated media to grow a
microorganism chosen are known as special media. It’s further categorized into different
groups:
• Selective media: It inhibits the growth of selected microorganisms while allowing the
other to flourish. Examples include desoxycholate citrate medium for dysentery bacilli
or mannitol salt agar containing 7.5% NaCl for Staphylococcus.
• Enriched media: It contains complex organic substances like hemoglobin, serum,

blood, or growth factors to support the growth of certain microbes. Examples are blood
agar (widely used to grow certain streptococci and other pathogens) and chocolate agar.
• Indicator media: It contains an indicator that changes color when a certain bacterium
grows on the medium. For example, the addition of sulfite in the Wilson and Blair
medium changes color to black when Salmonella typhi colonies grow on the medium.
• Differential media: This media allows the growth of different bacterial species and
distinguishes them based on their size, shape, color, or formation of gas bubbles or
precipitates in the medium. Examples are MacConkey medium and blood agar.
• Transport media: It’s a buffer solution containing peptone, carbohydrates, and other
nutrients (except growth factors) to maintain the viability of the bacteria during transport
without allowing their multiplication. An example is the Stuart medium for gonococci.
• Anaerobic media: It contains ingredients that support the growth of anaerobic bacteria.
An example is Robertson’s cooked meat media.
Now, the common culture techniques used in microbiology labs include:

Aseptic Technique

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Aseptic Techniques

• Aseptic technique is a procedure that is performed under sterile

• A fomite is any inanimate object or substance capable of

• Aseptic technique is essential in the microbiology laboratory to

• Flaming is one of the physical methods of sterilization and must

• This does not include bacterial endospores.

• The type of disinfectant to be used depends on the surface or material to be disinfected.

• Sanitization is applied in everyday life in order to control possible infections or spoilage of

• Often the sanitizer used is a compound such as soap or detergent.

Isolation is a microbiological technique in which a specific microbial strain is isolated from a

• Enriched media: It contains complex organic substances like hemoglobin, serum,

Now, the common culture techniques used in microbiology labs include:

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