Norman D.

Yasis BSN-1 Microbiology and parasitology

1. How are bacteria being cultured in the laboratory?

The basic procedure for culturing a bacterial species is pretty straightforward. Take a sample—
for example ocean water, soil, or spit—and dilute it in water. Then spread a droplet of this
dilution on a petri dish full of nutrients. Each individual bacterium lands in a unique spot on the
dish.
What are the protocols?
 Pick a single colony from a freshly streaked plate and inoculate a starter culture with 3
to 5 mL of media.
 Use the appropriate antibiotic and incubate at 37 °C* for approximately 8 hours while
shaking at 250-300 rpm.
 Use the starter culture to inoculate an overnight culture. Dilute the starter culture 1:500
to 1:1000 in a large flask with the appropriate volume of media and incubate at 37 °C*
for 12 to 16 hours while shaking at 250-300 rpm.
*This temperature is for standard E. coli. Some bacteria require different culture temperature*

2.
3. What are the Different Culture media in bacterial culture?
Types of Culture Media Based on Consistency/ Physical State
 Solid medium-It is for the isolation of bacteria as a pure culture on a solid
medium. Robert Koch realized the use of solid media.

 Semi Solid Medium-This media shows the motility of bacteria and the cultivation of
microaerophilic bacteria. This media has agar at a concentration of 0.5% or less. It has a
jelly consistency.

 Liquid Medium-This media shows the growth of a large number of bacteria.

It is called Broth that allows bacteria to grow uniformly with turbidity. The growth
occurs at 37ºC in an incubator for 24hrs.
Liquid media don’t have the addition of agar; it is for fermentation studies.

4. Draw Different kinds of streak plate method used in bacterial culture.

5. What is aseptic Technique?
Aseptic technique is a fundamental and important laboratory skill in the field of microbiology.
Microbiologists use aseptic technique for a variety of procedures such as transferring cultures,
inoculating media, isolation of pure cultures, and for performing microbiological tests. Proper
aseptic technique prevents contamination of cultures from foreign bacteria inherent in the
environment.

How important is aseptic technique in isolating microorganism?

Aseptic technique is of utmost importance to maintain pure stock cultures while transferring
cultures to new media. Aseptic technique is also essential for isolation of a single species of
microorganism from a mixed culture to obtain a pure culture.

6. What are the different physical and chemical methods in inhibiting the growth of
bacteria?

Physical Methods:
Heat: Heat can be used to kill bacteria by denaturing their proteins and disrupting their
cellular structures. Techniques like boiling, autoclaving, and pasteurization are commonly used
to apply heat for bacterial inhibition.
Filtration: Filtration involves passing a liquid or gas through a filter with tiny pores that trap
bacteria, preventing their passage through the filter.
Ultraviolet (UV) Radiation: UV radiation damages the DNA of bacteria, preventing them
from replicating and ultimately leading to their death.
Irradiation: Ionizing radiation, such as gamma rays or electron beams, can penetrate
materials to kill bacteria by damaging their DNA and cellular structures.
Desiccation: Removing water from the environment can inhibit bacterial growth since most
bacteria require water to survive and reproduce.
Osmotic Pressure: High osmotic pressure, achieved through the addition of salts or sugars,
can cause water to leave bacterial cells through osmosis, leading to dehydration and inhibition
of growth.

Chemical Methods:
Disinfectants: Chemicals such as bleach (sodium hypochlorite), hydrogen peroxide, alcohol,
and quaternary ammonium compounds are commonly used as disinfectants to kill bacteria on
surfaces.
Antiseptics: Similar to disinfectants, antiseptics are used on living tissues to kill or inhibit the
growth of bacteria. Examples include iodine, chlorhexidine, and alcohol-based hand sanitizers.
Antibiotics: Antibiotics are chemical substances produced by microorganisms or synthesized
in the laboratory that can kill or inhibit the growth of bacteria by targeting specific cellular
processes or structures.
Preservatives: Chemicals such as benzoic acid, ascorbic acid, and sodium nitrite are added
to food and other products to inhibit the growth of bacteria and other microorganisms, thereby
extending shelf life.
Metal ions: Certain metal ions such as silver, copper, and zinc have antimicrobial properties
and can be used to inhibit bacterial growth in various applications, including wound care and
water purification.

These methods can be used alone or in combination to effectively inhibit the growth of
bacteria in different environments and applications.
References:

 Microbial Growth Protocols (sigmaaldrich.com) &Bacteria Culture - Science in the News

(harvard.edu)
 Aseptic Technique and the Transfer of Microorganisms (Theory) : Microbiology Virtual
Lab I : Biotechnology and Biomedical Engineering : Amrita Vishwa Vidyapeetham Virtual
Lab
 Microbial Culture Media- Definition, Types, Examples, Uses (microbenotes.com)