INTRODUCTION TO MICROBIOLOGY

DEFINITIONS
SCIENCE --- study or collection of knowledge of natural events and matls in an orderly fashion MICROBIOLOGY - an advanced biology - study of very small living organisms called microorganisms or microbes 1. Characteristics of Microbes a. Ubiquitous - virtually everywhere b. Invisible to the naked eye c. Vast - these being approximately ten times as many microorganisms as the total # of cells that make up the human body

2. Types of Microbes A. non pathogens - do not cause disease 1. microbial allies - beneficial to the human body 2. those that have no effect at all B. 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

 Microorganisms

(Microbes)

87% Beneficial Microorganisms

3% Pathogens

10% Opportunists

PIONEERS IN THE SCIENCE OF MICROBIOLOGY

ANTON VAN LEEUWENHOEK (1632   

1723) Father of Microbiology, Father of Bacteriology, Father of Protozoology Made the single lens microscopes or simple microscopes. Discovered animalcules (live bacteria and protozoa) Spontaneous generation or abiogenesis arose from his findings--- life can arise spontaneously from nonliving materials.

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 cause specific infectious disease. Developed rabies vaccine in dogs and used it as a vaccine to treat human rabies.

ROBERT KOCH (1843-1910)

    

Gave significant contributions to the Germ Theory of disease Kochs postulates--- scientific steps Developed methods of fixing, staining and photographing bacteria. Developed methods of cultivating bacteria on solid media. Discovered Bacterium (Mycobacterium tuberculosis and Vibrio cholerae)

OTHER PIONEERS


Edward JennerLate 1700 --discovered 1st vaccine (used for smallpox) Joseph Lister (1865-1870)and Ignaz Semmelweiss (1847) --Aseptic techniques Paul Erlich (1890s to 1900) --Developed the 1st chemotherapeutic agent: Salvarsan vs. Syphilis.

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 immune response

APPLICATION OF MICROBIOLOGY
   

Indigenous microflora--- beneficial Opportunistic pathogenscolonize or inhabit bodies. MO involve in the decomposition of dead organisms---saprophytes MO cause 2 categories of diseases 1. Infectious disease pathogen colonizedisease. 2. Microbial intoxicationpathogentoxin humans ingest toxin---disease.

FUNCTIONAL CLASSIFICATION
 

GENERAL MICROBIOLOGY classification of MO & how they function. MEDICAL MICROBIOLOGYthe study of pathogens, the disease they cause and the bodys defenses against disease. VETERINARY MICROBIOLOGY the spread and control of infectious diseases among animals. AGRICULTURAL MICROBIOLOGY the beneficial and harmful role of microbes on the soil, plants, crops and foods. SANITARY MICROBIOLOGYpurification and processing of water supplies and processing and disposal of garbage and sewage wastes.

INDUSTRIAL MICROBIOLOGY growth and maintenance, research of microorganisms to produce commercial and pharmaceutical products. 1. Industrial microbiologists---produce commercial products. 2. Applied Microbiologists--- produce commercial products. ENVIRONMENTAL MICROBIOLOGY (MICROBIAL ECOLOGY) cycling of elements by microbial, environmental and geochemical processes. MICROBIAL PHYSIOLOGY AND GENETICS function of MO, structure and physiology.

TOOLS OF MICROBIOLOGY

DEFINITION
 

Tools and Techniques used to study microorganisms. Optical instrument - used to observe tiny objects that cannot be seen at all with the unaided human eye. Simple Microscope- microscope containing only one magnifying lens. Magnifying glass, Anton van Leeuwenhoek Compound Microscope- microscope that contains more than one magnifying lens. Compound light microscope. Hans Jansen and son Zacharias. Photomicrographs- photographs taken through the lens system of compound microscopes.

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

Illuminating Parts: - Parts that modify light that illuminate object of study - Abbe condenser: concentrates light - Mirror: reflects light or uses bulbs as main light source - Iris diaphragm: regulates the amount of light 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

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.

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)

II. STAINING PROCEDURES

OBJECTIVES: 1. Kill the organism 2. Preserve morphology 3. Anchor smear to slide A. Simple staining - Aqueous or alcohol solution of a single basic dye - Used to determine size, shape and morphological arrangement - Ex. Methylene blue

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

B. Differential staining- 2 or more dyes that may differentiate 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 Violet (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 A M a. Carbolfuchsin (Primary stain) b. Acid-alcohol (Decolorizer) c. Methylene Blue (Counterstain)

Gram staining Gram (+) Color Blue Pink

Gram (-) Pink Red

Peptidoglycan Thick Layer

Thin layer

Techoic acid in Present cell wall Lipopolysacch Absent aride in cell wall

Absent Present

C.Structural Stains - Observe capsules, spores, flagella 1. Negative stain - Used to demonstrate the presence of capsules - capsule (unstained halo) around bactl cells against dark background. 2. Endospore stain - Malachite green 3. Flagella stain - Carbolfuchsin