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Second year Lab Medicine (Females)
                 1433- 1434 A.H. / 2012- 2013
              Microbiology Practical (Course I)




• By:
        Hibah A. W. Abusulaiman
Once upon a time…

  Please don’t sleep
Fashionable friends
Abrar’s…
Sara started the preparation early
She got a luxurious manicure and
         padicure plus…
An elegant evening gown
Lubna started the preparation 2 days
        before the wedding
She got an elegant evening gown,
              but…
A poor manicure and padicure
Both attended the wedding in…
After 45 days, Lubna was suffering
              from…
Diagnosed as Hepatitis Patient
After discussion with the Doctor, she
knew it’s because of the poor hygienic
        manicure and pedicure
Good hygiene   Poor hygiene
Destruction of Microorganisms
So, do you think we are in a war
            with…?!
Simply because they are the
        agents of…
 Contamination, infection and decay
Early civilization
In the 1800s

 Aseptic techniques invented to
prevent contamination of surgical
            wounds
Many organizations classified infection
control systems and substances into:

• Physical methods.
• Chemical methods.
        Referred broadly as Germicides
Germicidal systems fall into three
              categories:


1. Decontamination.
2. Disinfection.
3. Sterilization.
1. Decontamination:
• Reduction of pathogenic microorganisms to be
  safe to handle without protective attire.
• The Lowest level of control.
• Includes physical cleaning with soap and
  detergents and removing of most organic and
  inorganic materials.
• It is the critical step toward disinfection and
  steilisation.
2. Disinfection:
• Disinfectants are liquid chemical agents that
  kill targeted pathogens but not spores . It can
  also be solid or gaseous.
• It is the next level of control.
3. Sterilization
• The complete elimination of viable organisms
  including spores.
• The highest level of control
Factors influencing disinfection
• Number of microorganisms.
• Species and type of microorganism.
• Temperature.
Physical                         Chemical
Antimicrobial                    Antimicrobial
   Agents                           agents


                Destruction of
                Microorganisms
Physical Antimicrobial Agents

               • Physical environmental
                 agents can destroy large
                 concentrations of
                 microorganisms in lab
                 cultures or a clinical
                 specimen.
Physical Antimicrobial Agents

                   Dry heat




    Moist heat                     Radiation

                     Physical
                   sterilization



    Incineration                    Filtration



                    Osmotic
                    pressure
Radiation




           UV Light

To inactivate microorganisms on
            surfaces
Filtration


        •Filtration is used to
        remove bacteria, fungi
        and their endospores
        from air or solutions.
Osmotic Pressure




• Extremely hypertonic conditions can cause
  plasmolysis.
Incineration
Autoclave
Moist heat- Steam sterilization
        121OC for 15 min
Dry heat
160OC for 2 hours OR 170OC for an hour
Chemical Antimicrobial Agents
• In considering their application to patient
  care, two general classes:
  1. Those destroying pathogenic microorganisms in
     the environment (disinfectants) or on skin
     (antiseptics).
  2. Those administered to patients for treatment of
     infectious diseases (antimicrobial agents, often
     called antibiotics).
Desinfectants
 Disinfectant    Bctericidal   Sporicidal   Fungicidal   Viricidal
  Ethyl and        Good          Poor         Good        good
  isopropyl
   alcohols
   Chlorine        Good          Good         Good        Good
 Disinfectants
Formaldehyde       Good          Good         Good        Good
     and
Glutaraldehyde
  Hydrogen         Good          Good         Good        Good
  peroxide
  Iodophors        Good          Poor         Good        Good
Evaluating a disinfectant

• Direct spray method.

• Testing the quality of disinfectant from
  different brands.
Testing Disinfectant from Different
                      Brands
                           0.5 mL




    5 mL Bacterial     5 mL Clorox   5 mL DAC   5 mL Gento   5 mL Dettol
Suspension in Nutrient
   Broth (Control)
Testing Disinfectant from Different
                         Brands




                                                   Water Bath

Bacterial Suspension in Nutrient                   Bacteria in DAC   Bacteria in Gento Bacteria in Dettol
        Broth (Control)       Bacteria in Clorox
Testing Disinfectant from Different
                      Brands
      2 Loopfulls




                                                                              48 hrs
    Water Bath                                                            Incubation
                                                                          in the 37O C
                             After 5min After 10 min in After 20 min in
Bacterial Suspension in    in Water Bath Water Bath      Water Bath
Nutrient Broth (Control)
Observe the bacterial growth
Objectives
•   Some infection control terms.
•   Factors influencing disinfectants.
•   Physical destruction of microorganisms.
•   Chemical destruction of microorganisms.
•   Methods of evaluating disinfectants.
Thank you for listening
   Any questions?!
Thank you for paying attention 

          Any questions?!

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Lab2 Destruction of microbes