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CONTROLLING MICROBIAL
GROWTH
INTRODUCTION TO MICROBIOLOGY
MUNIRA SHAHBUDDIN, PhD.
BTEN 2210
CHEMOTHERAPEUTICS
- Any drugs or agents used to
treat any disease
- The chemotherapeutic drugs
used to treat infectious
disease collectively referred
to as antimicrobial agents.
- An antibiotic is a substance
produced by a microorganism
that is effective in killing or
inhibiting the growth of other
microorganism
CONTROLLING MICROBIAL GROWTH IN
VITRO
Factors that affect microbial growth:
1. Availability of nutrient
2. Moisture
3. Temperature
4. pH
5. Light
6. Osmotic pressure and salinity
7. Barometric Pressure
8. Atmosphere
MICROORGANISMS
- CLASSIFICATION
Different type of microorganisms
Psychophilic
Mesophilic
Halophilic (prefer salty environment)
Haloduric
Alkaliphilic
Acidophilic
Piezophilic
CULTURING BACTERIA IN
LABORATORY
The earliest successful attempt to successfully
culture microorganisms in laboratory setting
were made by Ferdinand Cohn (1872), Joseph
Schoreter (1875) and Oscar Brefeld (1875).
Robert Koch describe his technique in 1881.
In 1878, Joseph Lister became the first person
to obtain the pure culture of bacterium
(Streptococcus lacti) in a liquid medium.
Koch, Pasteur and Lister made significant
contributions to the germ theory of disease.
CULTURING BACTERIA IN
LABORATORY
Bacterial growth – proliferation or
multiplication of bacteria.
Bacteria multiply by binary fission, which is
called organism’s generation time.
Microorganisms that are difficult to grow in
the laboratory are said to be fastidious.
CULTURING BACTERIA IN
LABORATORY
Selective medium: is used
to discourage the growth of
certain organisms without
inhibiting the growth of
organism being sought
Enriched medium: broth or solid
medium containing rich supply of
special nutrient that promotes the
growth of fastidious organisms.
INOCULATION OF CULTURE
MEDIA
In clinical microbiology laboratory, culture
media are routinely inoculated with clinical
specimens. Inoculation involves adding a
portion of the specimen to the medium using
sterile inoculating loop using a process
referred to as streaking.
ASEPTIC TECHNIQUE
It is practiced in microbiology laboratory to
prevent infection of individuals and
contamination of the work environment,
clinical specimen and cultures.
Unwanted organisms are referred to as
contamination
CONTAMINATION IN
CULTURE
INCUBATION: a chamber
containing appropriate
atmosphere and moisture
To culture human pathogen, the incubator is set at
35 – 37 oC.
There are three types of incubators in CML:
1. A CO2 incubator to maintain the concentration
of 5-10% CO2 – it contains O2 about 15-20%
2. A non CO2 incubator – only contain the room
air and O2 about 15-20%
3. An anaerobic incubator which is devoid of
oxygen.
BACTERIAL POPULATION
COUNT
Determination of total bacterial count
in the liquid
1. Spectrophotometer technique
2. Viable plate count
3. Haemacytometer
Coulter counter: Particle size analysisCell count using Haemacytometer
Cell count using Spectrophotometer
BACTERIA POPULATION
GROWTH CURVE
A bacterial growth curve consist of four phases
1. Lag (adaptation)
2. Log (exponential)
3. Stationary (Plateau)
4. Death
CULTURING INTRACELLULAR
PATHOGENS
Certain microbes can only
survive and multiply within
living cells.
- They can be propagated
in laboratory using
embryonated chicken
eggs, laboratory animals
or cell culture
CULTURING FUNGI IN
LABORATORY
Fungi including yeast, molds and
dimorphic fungi will grow on and in
various solid and liquid culture media.
There is no one medium that is best for
all medically important fungi.
Antibacterial agent often added to
inhibit/suppress the growth of
bacteria.
https://blog.mycology.cornell.edu/2008/01/10/a-simple-way-to-preserve-fungal-cultures/
CULTURING FUNGI IN
LABORATORY
Laboratory personnel must exercise
cautious when culturing fungi because
the spores of fungi are highly
infectious.
METHOD OF INHIBITING
GROWTH OF MICROBES IN
LABORATORY
METHODS OF
STERILIZATION:
1. Dry heat
2. Cold
3. Radiation
4. Filtration
5. Dessication
6. Autoclave (moist
heat)
Sterilization: Destruction of elimination
of all microbes
Disinfection: Elimination of most all
pathogens (except bacterial spores)
METHOD OF INHIBITING
GROWTH OF MICROBES IN
LABORATORY
Lyophilization: a process that combines
dehydration (drying) and freezing.
Disinfectant: the use of chemical
agents to inhibit the growth of
pathogen.
EFFECTIVENESS OF
ANTIMICROBIAL
PROCEDURES
Effectiveness
of
antimicrobial
procedures
Temperature
Concentration
Type of
microbes
Number of
microbes
Presence of
spores
Presence of
proteins in
feces, blood,
vomitus, pus
Time
CONTROVERSIES RELATING TO THE USE OF
ANTIMICROBIAL AGENTS IN ANIMAL FEED
AND HOUSEHOLD PRODUCTS
40% of the antibiotic manufactured in
the United States are used in animal
feed. The reason is obvious: to prevent
infections that could lead to economic
losses.
THE USE OF ANTIMICROBIAL
AGENTS IN EVERYTHING?
HYGENIC HYPOTHESIS?
Many scientists believe that children
must be exposed to all sorts of
microorganisms during their growth
and development so their immune
system will be developed correctly and
capable of properly responding to
pathogen in later years.

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Controlling microbial growth in vitro

  • 1. CONTROLLING MICROBIAL GROWTH INTRODUCTION TO MICROBIOLOGY MUNIRA SHAHBUDDIN, PhD. BTEN 2210
  • 2. CHEMOTHERAPEUTICS - Any drugs or agents used to treat any disease - The chemotherapeutic drugs used to treat infectious disease collectively referred to as antimicrobial agents. - An antibiotic is a substance produced by a microorganism that is effective in killing or inhibiting the growth of other microorganism
  • 3. CONTROLLING MICROBIAL GROWTH IN VITRO Factors that affect microbial growth: 1. Availability of nutrient 2. Moisture 3. Temperature 4. pH 5. Light 6. Osmotic pressure and salinity 7. Barometric Pressure 8. Atmosphere
  • 4. MICROORGANISMS - CLASSIFICATION Different type of microorganisms Psychophilic Mesophilic Halophilic (prefer salty environment) Haloduric Alkaliphilic Acidophilic Piezophilic
  • 5. CULTURING BACTERIA IN LABORATORY The earliest successful attempt to successfully culture microorganisms in laboratory setting were made by Ferdinand Cohn (1872), Joseph Schoreter (1875) and Oscar Brefeld (1875). Robert Koch describe his technique in 1881. In 1878, Joseph Lister became the first person to obtain the pure culture of bacterium (Streptococcus lacti) in a liquid medium. Koch, Pasteur and Lister made significant contributions to the germ theory of disease.
  • 6. CULTURING BACTERIA IN LABORATORY Bacterial growth – proliferation or multiplication of bacteria. Bacteria multiply by binary fission, which is called organism’s generation time. Microorganisms that are difficult to grow in the laboratory are said to be fastidious.
  • 7. CULTURING BACTERIA IN LABORATORY Selective medium: is used to discourage the growth of certain organisms without inhibiting the growth of organism being sought Enriched medium: broth or solid medium containing rich supply of special nutrient that promotes the growth of fastidious organisms.
  • 8.
  • 9. INOCULATION OF CULTURE MEDIA In clinical microbiology laboratory, culture media are routinely inoculated with clinical specimens. Inoculation involves adding a portion of the specimen to the medium using sterile inoculating loop using a process referred to as streaking.
  • 10. ASEPTIC TECHNIQUE It is practiced in microbiology laboratory to prevent infection of individuals and contamination of the work environment, clinical specimen and cultures. Unwanted organisms are referred to as contamination
  • 12. INCUBATION: a chamber containing appropriate atmosphere and moisture To culture human pathogen, the incubator is set at 35 – 37 oC. There are three types of incubators in CML: 1. A CO2 incubator to maintain the concentration of 5-10% CO2 – it contains O2 about 15-20% 2. A non CO2 incubator – only contain the room air and O2 about 15-20% 3. An anaerobic incubator which is devoid of oxygen.
  • 13. BACTERIAL POPULATION COUNT Determination of total bacterial count in the liquid 1. Spectrophotometer technique 2. Viable plate count 3. Haemacytometer
  • 14. Coulter counter: Particle size analysisCell count using Haemacytometer
  • 15. Cell count using Spectrophotometer
  • 16. BACTERIA POPULATION GROWTH CURVE A bacterial growth curve consist of four phases 1. Lag (adaptation) 2. Log (exponential) 3. Stationary (Plateau) 4. Death
  • 17. CULTURING INTRACELLULAR PATHOGENS Certain microbes can only survive and multiply within living cells. - They can be propagated in laboratory using embryonated chicken eggs, laboratory animals or cell culture
  • 18. CULTURING FUNGI IN LABORATORY Fungi including yeast, molds and dimorphic fungi will grow on and in various solid and liquid culture media. There is no one medium that is best for all medically important fungi. Antibacterial agent often added to inhibit/suppress the growth of bacteria. https://blog.mycology.cornell.edu/2008/01/10/a-simple-way-to-preserve-fungal-cultures/
  • 19. CULTURING FUNGI IN LABORATORY Laboratory personnel must exercise cautious when culturing fungi because the spores of fungi are highly infectious.
  • 20. METHOD OF INHIBITING GROWTH OF MICROBES IN LABORATORY METHODS OF STERILIZATION: 1. Dry heat 2. Cold 3. Radiation 4. Filtration 5. Dessication 6. Autoclave (moist heat) Sterilization: Destruction of elimination of all microbes Disinfection: Elimination of most all pathogens (except bacterial spores)
  • 21.
  • 22. METHOD OF INHIBITING GROWTH OF MICROBES IN LABORATORY Lyophilization: a process that combines dehydration (drying) and freezing. Disinfectant: the use of chemical agents to inhibit the growth of pathogen.
  • 23. EFFECTIVENESS OF ANTIMICROBIAL PROCEDURES Effectiveness of antimicrobial procedures Temperature Concentration Type of microbes Number of microbes Presence of spores Presence of proteins in feces, blood, vomitus, pus Time
  • 24. CONTROVERSIES RELATING TO THE USE OF ANTIMICROBIAL AGENTS IN ANIMAL FEED AND HOUSEHOLD PRODUCTS 40% of the antibiotic manufactured in the United States are used in animal feed. The reason is obvious: to prevent infections that could lead to economic losses.
  • 25. THE USE OF ANTIMICROBIAL AGENTS IN EVERYTHING?
  • 26. HYGENIC HYPOTHESIS? Many scientists believe that children must be exposed to all sorts of microorganisms during their growth and development so their immune system will be developed correctly and capable of properly responding to pathogen in later years.