Microbiology

UNIT I : Scope and History UNIT II : Microscopy and Staining UNIT III : Growth and Culturing UNIT IV : Sterilization and Disinfection UNIT V : The Bacteria UNIT VI : Viruses

Scope and History of Microbiology

Why Study Microbiology The Microbes Disease Causes by Microorganism History Roots

SCOPE AND HITORY OF MICRIBIOLOGY

Why Study Microbiology
Microorganisms are part of the human environment and are there fore important to human health and activities. The study of microorganisms provides insight into life processes in all forms of life.

Scope of Microbiology
The Microbes Microbiology is the study of all microorganisms (microbes) in the microscopic range. This include : bacteria,algae,fungi,viruses,and protozoa

Diseases caused by Microorganisme

Bacterial diseases: Syphilis,tetanus,trachoma,meningitis,etc Viral diseases : AID,Hepatitis,yellow fever,etc Protozoan disease : Malaria,Amebiasis Helminth diseases : Trichinosis.

The germ theory of disease

Pasteur further contributions

Kochs Contributions
Kochs Postulates: 1.The specific causative agent must be found in every case of the disease. 2.The disease organism must be isolated in pure culture 3.Inoculation of a sample of the culture into a healthy,susceptible animal must produce the same disease. 4.The disease organisms must be recovered from the inoculated animal.

II.Microscopy and Staining

Principles of microscopy Light Microscopy Electron Microscopy Principles of staining

MICROSCOPY
The technology of making very small things in visible to the human eye.

Comparison of Types of Microscopy

Metric Unit
Centimeter =0,01 m Millimeter =0,001 m Micrometer =0,000001 m Nanometer =0,000.000.001 m Angstrom = 0,000.000.000.1 m

III. Growth and Culturing of Bacteria

Culture Media Growth and Cell division Phases of Growth Measuring Bacterial Growth Factor Affecting Bacterial Growth Culturing Bacteria

CULTURE MEDIA

In nature, microorganims are growth on nature media or nutrients available in water,soil,and living or dead organic material.

In The Laboratory
Microorganisms are grown in synthetic media : 1.Defined synthetic media: consist of known quantities of spesific nutrien 2. Complex media : consist of nutrients of reasonably well known composition that vary in composition from batch to batch.

Commonly Used Media

Most routine laboratory cultures make us of peptones,or fish proteins. Other substances such as yeast extract,serum, whole blood, etc.

Diasnostic Media.

GROWTH AND CULTURING

1. Microbial growth can be defined as the orderly increase in quantity of all cell componen and in the number of cells of an organims 2.Because of limited in crease in cell size and the frequency of cell division, growth in microorganism is measured by increase in cells number.

Division Cell
Most cell division in bacteria occur by binary fission,in which the nuclear body divides and the cell form are transverse septum that separates the original cell into two cells. Yeast cell and same bacteria divide by budding, in which a small, new cell develops from the surface of an existing cell.

Growth and Cell Division

Phases of Growth

Phases of growth
Lag phase : Not increasing in cell number, metabolically active. Log phase : devide at exponential or logaritmic, rate and with a constant generation time.these properties can be used to calculate both the number of generation time and the generation. Stationary phase : The number of new cell produced equals the number of cell dying.The medium contains limited nutriens and contains toxic quantities of waste materials. Decline phase ; death phase : many cells lose their ability to devide and eventually die. A logaritmic decrease in the number of cell results.

Measuring bacterial growth

Growth can be measured by serial dilution. Growth also can be measure by direct microscopic count, the most probable number technique,filtration,observing or measuring products of metabolism, and obtaining the dry weight of cell.

The technikque of serial dilution

Methods of obtaining cultures

The streak plate method.

Spread plate method

Put 0,1 ml desired concentration(reached), pour into surface of pre-poured agar then spread with abent rod,next is incubated.Bacterial colonies appear only on surface.

pour plate method

1 ml of dilution (concentration reached) is mixed with 9 ml of melted agar. Mix throughly and pour entire petri dish. Cool to hardness and incubate. Same colonies appear on surface,many are bellow surface.

The streak plate method

A drop or bit of culture on a wire inoculating loop is lightly streaked across the top of the agar in region1.The loop is flamed,the plate is rotated,and a few organism are picked up from region 1 and streaked out into region 2,The loop is flames again and the process is repeated in region 3.

Principles of staining
A stain or dye is a molecule that can bind to a structure and give it color. Most microbial stain are cationic (positively charged),or basic, dyes,such as methylene blue.

Comparison of Staining techniques

The Gram Stain

The gram stain, was devised by Hans christian Gram, in 1884. The gram stain probabily the most frequently used deferential stain. Gram was testing new methods of staining biopsy and autopsy material.

Steps in gram staining

Factors affecting bacterial growth.

1. Physical factor: pH, Temperature, Oxygen
2. Nutritional factor :carbon sources. Nitrogen sources, vitamin.

Physical factors
A.Optimum pH Bacteria are clasified as 1. Acidophiles :pH below 5,4. Exm :Lactobacilus 2. Neutrophiles (pH 5,4 -8,5). Ex : Bacteria that causes disease in human. 3.Alkaliphiles : pH 7,0-11,5. Ex :Agrobacterium,alcaligenes faecalis.

B.Temperature
Bacteria are clasified as :

1.Psychrophiles( 150C-200C ) a. Obligate psychrophiles (cannot grow above 200C ).Ex.Bacillus globisporus. b.Facultative psychophiles (grow best below 200C). Ex.Xanthomonas sp

 2. Mesophiles (250C 400C) 3.Thermophiles(500C -600C) Ex. Bacillus stearothermophilus. C.Oxygen a.aerobes (require oxygen to grow). Ex:pseudomonas b.anaerobes(do not requere it). Ex:Bacteroides.

Nutritional factors
Carbon sources Autotrophs : us CO2 as carbon source Heterotroph :require glocose or organic carbon source .Nitrogen sources : sulfur,phosphorus,Iron,etc. .Sulfur and phosphorus :Amino acid,organic phosphate . .Vitamin :B12, K (small amount or as a coenzyme.

Sterilization and Disinfection

1.Principles of Sterilization and Disinfection 2.Chemical antimicrobial agents
a.potency of chemical agent b.Mechanisms of action c.specific chemical agent

3. Physical antimicrobial agents.

Sterilization

Refers to the killing or removal of all organisms in any material or on object.

Disinfection

Refers to the reduction in numbers of pathogenic organism on obyects or in materials so that the organisms no longer pose a disease threat.

TERM OF RELATED TO STERILIZATION AND DISINFECTION

ANTISEPTIC : A chemical agent that can safely be used externally on living tissue to destroy microorganisms or to inhibit their growth Disinfectant : A chemical agent used on inanimate obyects to destroy migroorganisms . Most disinfectant do not kill spores.

 SANITIZER :A chemical agent typically used on food-handling equipment and eating utensils to reduse bacterial numbers so as to meet public health standards. Sanitization may simply refer to thorough washing with only soap or detergent. Bactericide : An agent that kills bacteria.

POTENCY OF CHEMICAL AGENT

The potency of chemical agent is affected by time, temperature, pH, Concentration of the agent.

Mechanisms of action of chemical agent

Action of chemical agent antimicrobial agent can be grouped according to their effects on protein, cell membranes and other cell components. Reactions that alter protein include hydrolysis, oxidation,and attachment of atoms or chemical geoups to protein molecules. Such reaction denature protein, rendering them nonfungtional. Reactions of other chemical agents damage nucleic acids and energy-producing systems. Demage to nucleic acids is an important means of inactivating viruses.

SPECIFIC CHEMICAL ANTIMICROBIAL AGENTS

Soaps and detergents aid in the removal of microbes,oils,and dirt but do not sterilize. Among the agent containing heavy metals, silver nitrate is used to kill gonococci, and mercury containing compounds are used to disinfect instrument and skin.

 Acid are commonly used as food preservatives, alkali in soap helps destroy microorganisms. Alkohols are used to disinfectants Among the agents containing halogens, chlorine is use to kill pathogens in water, and iodine is major ingredient in several skin disinfectants.

 Phenol derivatives can be used on skin, instruments,dishes, and furniture, and to destroy discarded cultures they work well in the presence of organic materials. Oxidizing agents are particularly useful in disinfecting puncture wounds.

 Alkylating agents can be used to disinfect or sterilize a variety of materials, but all are carcinogens. Same dyes, plant oils, sulfur containing subtances and nitrates can be used as disinfectans or food preservatives.

PHYSICAL ANTIMICROBIAL AGENTS

Heat destroys microorganisms by denaturing protein, melting lipids,and when open flame is used,by incineration.

Dry heat,Moist heat,Pasteurization

Dry heat is used to sterilize metal obyects and glassware. Flame is used to sterilize inoculating loops and the mouths of culture tubes. Autoclave uses moist heat underpresure, is a common instrument for sterilization and is very efective when proper procedures are followed. Pasteurization kill most patogens milk,does not sterilize.

Refrigeration, freezing,drying,and Freeze-drying.

Can be used to retard the growth of microorganisms.

Radiation
Used to control microorganisms includes ultraviolet light, ionizing radiation,and some times microwaves and strong sunlight.

Sonic and Ultra sonic waves

Can kill microorganisms but are used mostly for sonication,the disruption of cell by sound waves

Filtration
Can be used to sterilize subtances that are destroyed by heat, to separate viruses, and to collect microorganisms from air and water samples.

Osmotic pressure
High concentrations of sugar or salt create osmotic presure that plasmolisis of cell and prevents growth of microorganisms in gightly sweetened or salted foods.