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INTRODUCTION TOMICROBIOLOGY
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DEFINITIONSSCIENCE --- study or collection of knowledge of
natural events and matls in an orderly fashion
MICROBIOLOGY
- an advanced biology
- study of very small living organisms calledmicroorganisms or microbes
1. Characteristics of Microbes
a. Ubiquitous - virtually everywhere
b. Invisible to the naked eyec. Vast - these being approximately tentimes as many microorganisms as thetotal # of cells that make up the humanbody
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2. Types of Microbes
A. non pathogens - do not cause disease1. microbial allies - beneficial to the human
body
2. those that have no effect at allB. pathogens - cause disease
1. infectious disease - pathogen colonizes the
human body then creates disease
2. microbial intoxication pathogen produces
a toxin in vitro person ingest the toxin
toxin causes the disease
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Microorganisms (Microbes)87% Beneficial Microorganisms
3% Pathogens 10% Opportunists
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PIONEERS IN THE SCIENCE OF
MICROBIOLOGYANTON VAN LEEUWENHOEK (1632-
1723)
Father of Microbiology, Father ofBacteriology, Father of Protozoology
Made the single lens microscopes or simplemicroscopes.
Discovered animalcules (live bacteria andprotozoa)
Spontaneous generation or abiogenesisarose from his findings--- life can arisespontaneously from nonliving materials.
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LOUIS PASTEUR (1822-1895)
French chemist Discovered alcoholic fermentation.
Process of alcoholic fermentation.
Disproved the Spontaneous generation or
Abiogenesis. Introduced terms aerobes and anaerobes
Pasteurization and sterilization
Discovered the cause of silkworm diseases.
Germ theory of Disease specific MO causespecific infectious disease.
Developed rabies vaccine in dogs and usedit as a vaccine to treat human rabies.
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ROBERT KOCH (1843-1910) Gave significant contributions to the Germ
Theory of disease
Kochs postulates--- scientific steps
Developed methods of fixing, staining andphotographing bacteria.
Developed methods of cultivating bacteria
on solid media.
Discovered Bacterium (Mycobacteriumtuberculosis and Vibrio cholerae)
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OTHER PIONEERS
Edward JennerLate 1700
--discovered 1st vaccine (used for smallpox)
Joseph Lister (1865-1870)and IgnazSemmelweiss (1847)
--Aseptic techniques
Paul Erlich (1890s to 1900)
--Developed the 1st chemotherapeutic agent:
Salvarsan vs. Syphilis.
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BRANCHES OF MICROBIOLOGY
Bacteriology the study of Bacteria
Phycology the study of Algae
Mycology the study of Fungi Protozoology the study of protozoa
Virology the study of viruses
Immunology- the study of immuneresponse
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APPLICATION OF MICROBIOLOGY
Indigenous microflora--- beneficial
Opportunistic pathogenscolonize or inhabitbodies.
MO involve in the decomposition of deadorganisms---saprophytes
MO cause 2 categories of diseases
1. Infectious disease pathogen
colonizedisease.2. Microbial intoxicationpathogentoxinhumans ingest toxin---disease.
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FUNCTIONAL CLASSIFICATION
GENERAL MICROBIOLOGY classification of
MO & how they function. MEDICAL MICROBIOLOGYthe study of
pathogens, the disease they cause and thebodys defenses against disease.
VETERINARY MICROBIOLOGY the spreadand control of infectious diseases amonganimals.
AGRICULTURAL MICROBIOLOGY the
beneficial and harmful role of microbes on thesoil, plants, crops and foods.
SANITARY MICROBIOLOGYpurification andprocessing of water supplies and processing
and disposal of garbage and sewage wastes.
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INDUSTRIAL MICROBIOLOGY growth and
maintenance, research of microorganisms to
produce commercial and pharmaceuticalproducts.
1. Industrial microbiologists---produce
commercial products.
2. Applied Microbiologists--- produce
commercial products.
ENVIRONMENTAL MICROBIOLOGY
(MICROBIAL ECOLOGY) cycling of elementsby microbial, environmental and geochemical
processes.
MICROBIAL PHYSIOLOGY AND GENETICS
function of MO, structure and physiology.
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TOOLS OFMICROBIOLOGY
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DEFINITION Tools and Techniques used to study
microorganisms. Optical instrument - used to observe tiny
objects that cannot be seen at all with theunaided human eye.
Simple Microscope- microscope containingonly one magnifying lens. Magnifying glass,
Anton van Leeuwenhoek
Compound Microscope- microscope that
contains more than one magnifying lens.Compound light microscope. Hans Jansen andson Zacharias.
Photomicrographs- photographs taken through
the lens system of compound microscopes.
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PARTS - COMPOUND MICROSCOPE Magnifying Parts:
-To enlarge objects of study
- objectives:
- Eyepiece or ocular objective: variable
magnification- Scanner: 5x magnification, used to study
larger organisms
- Low Power Objective (LPO): 10x
magnification
- High Power Objective (HPO): 40x
magnification
- Oil Immersion Objective: 100x magnification
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Illuminating Parts:
- Parts that modify light that illuminate object of
study- Abbe condenser: concentrates light
- Mirror: reflects light or uses bulbs as mainlight source
- Iris diaphragm: regulates the amount oflight that hits the object of study
Mechanical Parts:
- Supports the magnifying and illuminating parts
- Used to focus the lenses
- Draw tube, body tube, revolving nosepiece,dust shield, arm, stage, stage clips, coarse
adjustment knob, base, inclination joint
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I. TYPES OF MICROSCOPE
A. VISIBLE LIGHT MICROSCOPY
1. Bright-Field Microscope
---observe morphology of the organisms.
---does not resolve very small specimens(viruses)
2. Dark-field Microscope
---Dark background, light organisms
---used to detect Syphilis (Treponema
pallidum)3. Phase-Contrast Microscope
---Observe dense structures
---To facilitate detailed examination of the
internal structures of living specimens.
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4. Fluorescent Microscope
---Ultraviolet light
---used to show antibodies
B. ELECTRON MICROSCOPE
1. Transmission Electron Microscope (TEM)
---Highest magnification (10,000-100,000x)
---Cellular ultra structure and viruses
---2-D image
2. Scanning Electron Microscope (SEM)---Surface structure of cells and viruses
---3-D image
---magnification: (1000-10,000x)
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II. STAINING PROCEDURES
OBJECTIVES:
1. Kill the organism
2. Preserve morphology
3. Anchor smear to slideA. Simple staining
- Aqueous or alcohol solution of a singlebasic dye
- Used to determine size, shape andmorphological
arrangement
- Ex. Methylene blue
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Simple Staining Based on shape:
- Cocci: round or spherical bacteria
- Bacilli: rod-shaped or cigar shaped
bacteria
- Spirals: coiled bacteria- Spirillum: flexible coiled bacteria
- Spirochetes: rigid, coiled bacteria
Based on Arrangement of cells:
- Strepto: bacteria in chains
- Staphylo: bacteria in clusters
- Diplo: bacteria in pairs
- Tetra: bacteria in 4s
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B. Differential staining- 2 or more dyes that maydifferentiate one type of organism from one another.
1. Gram Stain- used to classify MO
- Dr. Hans Christian Gram (1884) ----- V I A S
a. Crystal VViolet (primary stain)
b. Grams Iodine (Mordant)
c. Alcohol (Decolorizer)
d. Safranin (Counterstain)
2. Acid-Fast Stain (Ziehl-Nielsen)
- Binds strongly to the bacteria that have a waxy
material in their cell wall
- Used to identify Mycobacterium, Nocardia
- Procedure: C AMa. Carbolfuchsin (Primary stain)
b. Acid-alcohol (Decolorizer)
c. Methylene Blue (Counterstain)
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C.Structural Stains
- Observe capsules, spores, flagella1. Negative stain
- Used to demonstrate the presence of
capsules
- capsule (unstained halo) around bactl
cells against dark background.
2. Endospore stain
- Malachite green3. Flagella stain
- Carbolfuchsin