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    Basic Microbiology

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    Masood Ahmed
    Microbiology is the study of microorganisms that cannot be seen with the naked eye. Microorganisms, also called germs, include pathogens that cause disease as well as many that are harmless or beneficial to the human body. The growth and survival of microorganisms depends on factors like temperature, pH, moisture, food availability, and oxygen levels. Methods to destroy microorganisms include antiseptics, disinfectants, and sterilization techniques using heat, chemicals, or radiation. Major types of microbes include bacteria, fungi, protozoa, parasites, and viruses. Gram staining is a common differential staining technique that classifies bacteria as either gram positive or gram negative based on differences in their cell wall

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    Basic Microbiology

    Uploaded by

    Masood Ahmed
    13 views17 pages
    Microbiology is the study of microorganisms that cannot be seen with the naked eye. Microorganisms, also called germs, include pathogens that cause disease as well as many that are harmless or beneficial to the human body. The growth and survival of microorganisms depends on factors like temperature, pH, moisture, food availability, and oxygen levels. Methods to destroy microorganisms include antiseptics, disinfectants, and sterilization techniques using heat, chemicals, or radiation. Major types of microbes include bacteria, fungi, protozoa, parasites, and viruses. Gram staining is a common differential staining technique that classifies bacteria as either gram positive or gram negative based on differences in their cell wall

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    Basic microbiology

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    Basic microbiology

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    Microbiology is the study of microorganisms that cannot be seen with the naked eye. Microorganisms, also called germs, include pathogens that cause disease as well as many that are harmless or beneficial to the human body. The growth and survival of microorganisms depends on factors like temperature, pH, moisture, food availability, and oxygen levels. Methods to destroy microorganisms include antiseptics, disinfectants, and sterilization techniques using heat, chemicals, or radiation. Major types of microbes include bacteria, fungi, protozoa, parasites, and viruses. Gram staining is a common differential staining technique that classifies bacteria as either gram positive or gram negative based on differences in their cell wall

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    © All Rights Reserved
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    Download as pdf or txt
    13 views17 pages

    Basic Microbiology

    Uploaded by

    Masood Ahmed
    Microbiology is the study of microorganisms that cannot be seen with the naked eye. Microorganisms, also called germs, include pathogens that cause disease as well as many that are harmless or beneficial to the human body. The growth and survival of microorganisms depends on factors like temperature, pH, moisture, food availability, and oxygen levels. Methods to destroy microorganisms include antiseptics, disinfectants, and sterilization techniques using heat, chemicals, or radiation. Major types of microbes include bacteria, fungi, protozoa, parasites, and viruses. Gram staining is a common differential staining technique that classifies bacteria as either gram positive or gram negative based on differences in their cell wall

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    Prashant Goel

    Manager Microbiology
     Microbiology is the science that
    studies living organisms that cannot
    be seen with the naked eye
    (microorganisms or microbes).

     Microorganisms are commonly


    known as germs.
     Pathogens: microorganisms that cause disease.
     most microorganisms are harmless.
     some microorganisms found in the body are
    beneficial to us.
     when a microorganism enters a part of the
    body other than where it is intended to be, it
    can be harmful.
     eg. E. coli from the colon (breaks down waste/makes
    vitamin K) can enter the urinary tract and cause an
    infection
     5 factors affect the growth of microorganisms:
     Temperature Require for Microorganism Growth.
     pH (only a few live in acidic environments)
     moisture
     food
     oxygen
     Above factors must be favorable if microorganisms
    are to live.
     Different microorganisms require different
    conditions for survival
     eg. aerobic microorganisms grow best with high levels of
    oxygen, while anaerobic microorganisms grow best in the
    absence of oxygen
     3 common methods destroy microorganisms:
     Antiseptics (eg. alcohol and betadine)
     used on outer surface like skin.
     slow or prevent the growth of pathogens.
     Disinfection (eg. Fungisicidal & Bacterisicidal)
     Not used on skin because irritation may result.
     Sterilization
     Autoclaves use steam under pressure to kill all
    microorganisms .
     Other methods include the use of chemicals, radiation,
    and gas sterilization.
     5 major types are:
     Bacteria
     Fungi
     Protozoa (protists)
     Parasites (rickettsiae)
     Viruses
     Most familiar of the microbes that infect
    humans.
     Classified by their shape and
    arrangement:
    Type Shape Arrangement

    Diplococci Round in pairs

    Staphylococci Round in groups or


    clusters
    Streptococci Round in chains

    Bacilli Rod-shaped in pairs, single, or


    in chains
    Spirilla Cork-screw or single
    spiral
     Gram Staining is the common, important, and
    most used differential staining techniques in
    microbiology, which was introduced by Danish
    Bacteriologist Hans Christian Gram in 1884.
    This test differentiate the bacteria into Gram
    Positive and Gram Negative Bacteria, which
    helps in the classification and differentiations
    of microorganisms
     When the bacteria is stained with primary stain
    Crystal Violet and fixed by the mordant, some
    of the bacteria are able to retain the primary
    stain and some are decolorized by alcohol. The
    cell walls of gram positive bacteria have a thick
    layer of protein-sugar complexes called
    peptidoglycan and lipid content is low.
     Decolorizing the cell causes this thick cell wall
    to dehydrate and shrink, which closes the pores
    in the cell wall and prevents the stain from
    exiting the cell. So the ethanol cannot remove
    the Crystal Violet-Iodine complex that is bound
    to the thick layer of peptidoglycan of gram
    positive bacteria and appears blue or purple in
    colour.
     In case of gram negative bacteria, cell wall also
    takes up the CV-Iodine complex but due to the
    thin layer of peptidoglycan and thick outer
    layer which is formed of lipids, CV-Iodine
    complex gets washed off. When they are
    exposed to alcohol, decolorizer dissolves the
    lipids in the cell walls, which allows the crystal
    violet-iodine complex to leach out of the cells.
    Then when again stained with safranin, they
    take the stain and appears red in color.
     Crystal Violet, the primary stain
     Iodine, the mordant
     A decolorizer made of acetone and alcohol
    (95%)
     Safranin, the counterstain
     Take a clean, grease free slide.
     Prepare the smear of suspension on the clean
    slide with a loopful of sample.
     Air dry and heat fix
     Crystal Violet was poured and kept for about
    30 seconds to 1 minutes and rinse with water.
     Flood the gram’s iodine for 1 minute and wash
    with water.
     Then ,wash with 95% alcohol or acetone for
    about 10-20 seconds and rinse with water.
     Add safranin for about 1 minute and wash
    with water.
     Air dry, Blot dry and Observe under
    Microscope.
     Gram Positive: Blue/Purple Color
    Gram Negative: Red Color
     Gram Positive Bacteria: Actinomyces, Bacillus,
    Clostridium, Corynebacterium, Enterococcus,
    Gardnerella, Lactobacillus, Listeria, Mycoplasma,
    Nocardia,,etc.
    Gram Negative Bacteria: Escherichia coli (E.
    coli), Salmonella, Shigella, and other
    Enterobacteriaceae, Pseudomonas,Moraxella,
    Helicobacter, etc
    THANK YOU

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