 Antoni Van Leeuwenhoek Is A ‘Father Of
Microbiology’ & ‘Father Of Microscopy.’
 Robert Koch (1834-1910) Is Also Known As The
'Father Of Bacteriology‘.
 Louis Pasteur Is The Father Of ‘Pasteurization’.
 Louis Pasteur Is Also Known As The Father Of
‘Modern Microbiology’.
 Elie Metchnikoff (1845-1916) Is Known As The
‘Father Of Phagocytosis’.
 Paul Ehrlich (1854-1915) Is Known As The
‘Father Of Chemotherapy’.
 Hans Christian Gram (1853-1933) Invented
Famous Bacterial Staining Procedure ‘Gram
Stain’.
 Micro-organism Or Microbe:- “An Organism That Can
Be Seen Only Through A Microscope. Microorganisms
Include Bacteria, Protozoa, Algae, And Fungi”.
 Classification of Micro-organisms:-
• Micro-organisms are classified into ten groups:
1. Bacteria
2. Viruses
3. Fungi
4. Protozoa
5. Rickettsiales
6. Algae
7. Moulds
8. Yeast
9. Chlamydia
10. Mycoplasma
unit-2micro-organisms-.pptx for students
 Classification of Micro-organisms:-
• Classification of microorganism depending on cell
type.
1) Eukaryotic micro-organisms
2) Prokaryotic micro-organisms
3) Non cellular micro-organisms
 Eukaryotic micro-organisms:- They have
complex cellular structure similar to human &
animal cell. Eg. Fungi, protozoa, algae
 Prokaryotic micro-organisms:- they are simple,
unicellular organisms having primitive nuclei &
mitochondria. Unicellular also known as single-
cell organisms. They are capable to live
independent.
 Prokaryotic micro-organisms:- they are simple,
unicellular organisms having primitive nuclei &
mitochondria. Unicellular also known as single-
cell organisms. They are capable to live
independent. Eg. Bacteria.
 Non cellular micro-organisms:- They are without
any cell forms called as acellular. Eg. Viruses.
unit-2micro-organisms-.pptx for students
 Structure of Micro-Organisms:-
• Bacteria are the most well known micro-
organisms.
• Bacteria are microscopic, prokaryotic, unicellular
organisms found everywhere.
 Size of the Bacteria:-
• Bacteria are very small in size. The unit of
measurement of bacteria is called micrometer
(mm).
• Bacteria generally measure 0.2-1.5mm in diameter
& 3.5mm in length.
 Morphology Of Bacteria
 Structure of Bacteria:-
 Morphology Of Bacteria:- Bacteria Is A
Prokaryotic, Unicellular Organism. Bacteria Are
Found Everywhere, They Are The Most Abundant
Living Organisms.
 Parts Of Bacteria:-
• Bacterial Cell Wall
• Capsule
• Cytoplasmic Membrane
• Cytoplasm
• Ribosomes
• Mesosomes
• Nucleus
• Flagella
• Bacterial cell wall:- It is complex rigid structure. It
gives bacteria its definite shape. It is semi-
permeable. The cell wall of gram-positive bacteria
is thicker than the gram-negative.
• Capsule:- It is a protective covering around the
bacterial cell wall. If this capsule is thick it’s called
capsule. If this layer is thin it’s called slime layer.
Capsule protects the bacteria from antibacterial
agents.
• Cytoplasmic membrane:- It is also called cell
membrane. It is thin & elastic. It is semi-permeable.
This membrane carry many enzymes & pigments.
• Cytoplasm:- It is viscous watery solution. It
contains variety of organic & inorganic solutes &
ribosomes.
• Ribosomes:- It is 10-20nm in diameter. Function of
this is protein synthesis.
• Mesosomes:- It is convoluted membranous bodies.
• Nucleus:- The bacterial nucleus does not possess
nuclear membrane & nucleolus. The genetic
information is contained in a single circular, double
strand molecule of DNA.
• Flagella:- It is long, hollow filaments. They are 10-
20nm in diameter & 3-20mm in length. They are
found on large number of bacteria. Flagella helps in
motility.
• Pili or Fimbriae:- It is hair like structure. They are
thinner & shorter than flagella. They allow
attachment of bacterial cell to other bacterial cell
surfaces.
• Flagella Types:- There are 4 types of
arrangements of flagella.
• 1. Monotrichous:- Single polar flagellum.
• 2. Lophotrichous:- Tuft or group of flagella at one
pole.
• 3. Amphitrichous:- Single flagella at both poles.
• 4. Cephalotrichous:- Tuft or group of flagella at
both poles.
• 5. Peritrichous:- Flagella all around the cell.
• 6. Atrichous:- An absence of flagella.
• Flagella Types:-
 Shape of the Bacteria:-
• Individual Bacteria Can Assume One Of 3 Basic
Shapes: Spherical (Coccus), Rodlike (Bacillus),
Or Curved (Vibrio, Spirillum, Or Spirochete).
 Cocci:- They are round or oval in shape. They
exist in various patterns.
• Types of Cocci:-
• Diplococci:- Occurs in pairs, Eg. Gonococci.
• Streptococci:- Occurs in chains.
• Staphylococci:- Occurs in clusters
• Tetrads:- Occurs in Group of 4.
• Sarcina (Octads):- Occurs in Group of 8.
unit-2micro-organisms-.pptx for students
 Bacilli (Bacillus):- They are rod or stick in shape.
It is also called a bacilliform bacterium. They
exist in various patterns.
•Types of Bacilli:-
•Diplobacillus:- Two bacilli arranged side by side
with each other.
•Streptobacillus:- Occurs in chains.
•Coccobacillus:- Oval and similar to Coccus.
unit-2micro-organisms-.pptx for students
 Spirilla (Spirillum):- They Are Non-flexible Spiral
Forms. They Have One To Three Fixed Curves In
Their Body. Eg. Spirillum Volutans.
 Vibrio:- They Are Curved Or Comma Shaped
Rods. Most Of Them Are Non-pathogenic But
Some Are Pathogenic. Eg. Vibrio Cholera
 Spirochaetes:- They Are Slender Spiral Filaments.
They Are Flexible. They Live In Soil & Stagnant
Pools. Eg. Treponema Pallidum
unit-2micro-organisms-.pptx for students
 Bacterial Growth:- Bacteria Are Unicellular Organisms That
Tend To Reproduce Asexually By The Means Of Binary
Fission.
• Bacterial Growth Is The Increase In The Number Of Bacterial
Cells Rather Than The Increase In Their Cell Size.
• The Growth Of These Bacterial Cells Takes Place In An
Exponential Manner, i.e., One Cell Divides Into 2, Then 4,
Then 8, 16, 32 And So On.
• The Time Taken For A Bacterial Cell To Double Is Called
‘Generation Time.’
• The Generation Time Varies Among Different Species Of
Bacteria Based On The Environmental Conditions They
Grow In.
• Clostridium Perfringens Is The Fastest Growing Bacteria
That Has A Generation Time Of 10 Minutes.
• Escherichia Coli Has A Doubling Time Of 20 Minutes.
• Mycobacterium Tuberculosis Is One Of The Slowest
Growing Bacteria, Taking About 12 To 16 Hours To
Double.
 Growth Curve:-
• In A Closed System With Enough Nutrients, A Bacteria
Shows A Predictable Growth Pattern That Is The Bacterial
Growth Curve.
• The Bacterial Growth Curve Shows The Preparation,
Division, Growth And Death Of The Bacterial Cells.
• The Bacterial Growth Progresses In Four Phases Namely –
Lag Phase, Log Phase, Stationary Phase And Death Phase.
• Bacteria Require Optimum Temperature, Ph, Moisture,
Oxygen, Carbon Source, Nitrogen Source And Other
Nutrients
unit-2micro-organisms-.pptx for students
• 1. Lag Phase:-
• The Bacteria Upon Introduction Into The Nutrient Medium
Take Some Time To Adapt To The New Environment.
• In This Phase, The Bacteria Does Not Reproduce But
Prepares Itself For Reproduction.
• The Cells Are Active Metabolically And Keep Increasing
In Size.
• The Cells Synthesise RNA, Growth Factors And Other
Molecules Required For Cell Division.
• 2. Log Phase:-
• Soon after the lag phase, i.e., the preparation phase, the
bacterial cells enter the log phase.
• This phase is also known as the exponential phase.
• This phase is marked by the doubling of the bacterial cells.
The cell number increases.
• The log phase continues until there is depletion of
nutrients in the setup.
• The stage also comes to a stop if toxic substances start to
accumulate, resulting in a slower growth rate.
• The cells are the healthiest at this stage and researchers
prefer to use bacteria from this stage for their experimental
processes.
• 3. Stationary Phase:-
• In The Stationary Phase, The Rate Of Growth Of The Cells
Becomes Equal To Its Rate Of Death.
• The Rate Of Growth Of The Bacterial Cells Is Limited By
The Accumulation Of Toxic Compounds And Also
Depletion Of Nutrients In The Media.
• The Cell Population Remains Constant At This Stage.
• 4. Death Phase:-
• This Is The Last Phase Of The Bacterial Growth.
• At This Stage, The Rate Of Death Is Greater Than The
Rate Of Formation Of New Cells.
• Lack Of Nutrients, Physical Conditions Or Other Injuries
To The Cell Leads To Death Of The Cells.
 Factors Influencing Growth Of Bacteria
Or Microbes:-
1. Nutrition
2. Moisture
3. Temperature
4. pH
5. Oxygen
6. Carbon Dioxide
7. Light & Other Radiations
8. Osmotic Effect
9. Mechanical & Sonic Stresses
1. Nutrition:- The Growth Of Microbe Depend On Adequate
Supply Of Suitable Nutrients Like Carbon, Nitrogen,
Phosphates, Ions & Metals.
2. Moisture:- It Is Very Essential For The Growth Of
Bacteria Because 80% Of The Body Of Bacteria Is Made Up
Of Water. Moist Places Are More Viable Media For Growth
Of Bacteria.
3. Temperature:- Bacteria Requires Specific Temp. Range
For Their Growth. Bacteria Can Be Killed By Very High
Temp. & Very Low Temp. Favorable Range Is 20 - 40 C.
ͦ ͦ
4. pH:- Some Bacteria Grow In Neutral Media, Some Grow In
Alkaline Media & Some Grow In Acidic Media.
• Acidophiles:- Organisms grow at pH between 0.5-6.
• Neutrophiles:- Organisms grow best at pH 7.
• Alkaliphiles:- Organisms grow best under at pH above 7.
5. Oxygen:- The Bacteria Which Can Grow Only In The
Presence Of Oxygen Are Known As ‘Aerobic Bacteria’. This
Bacteria Requires Oxygen For Life & Reproduction.
• Some Bacteria Can Grow In The Absence Of Oxygen Are
Known As ‘Anaerobic Bacteria’. This Bacteria Does Not
Requires Oxygen For Life & Reproduction.
6. Carbon Dioxide:- Some Bacteria Requires Extra CO2 In The
Air For Their Growth & Reproduction.
7. Light & Other Radiations:- Bacteria Grow In Dark
Places. Bacteria Can Be Destroyed Or Killed By UV Rays,
Ionizing Radiations, Infrared Rays & Other Radiations.
8. Osmotic Effect:- Osmotic Pressure Changes Can Effect
The Growth Of Bacteria. Sudden Changes In Osmotic
Pressure Can Cause Destruction Of Bacteria.
9. Mechanical & Sonic Stresses:- Bacteria Cell Wall Can Be
Ruptured & Bacteria Can Be Killed By Vigorous Shaking &
Ultrasonic Vibrations.
 Normal Flora:-
• The Normal Flora Are Micro-organisms Which Are Found
In Or On Our Body Without Causing Any Disease.
• Normal Flora Arranged In Two Groups:
1. Resident Flora:- It Is Consist Of Permanent
Microorganisms Which Are Regularly Found In A Given
Area.
2. The Transient Flora:- It Is Consist Of Pathogenic Or
Non-pathogenic Organisms That Stay On The Skin Or
Mucus Membranes For Hours, Day Or Week. They Do
Not Stay Permanently On The Surface.
• The most common sites of body inhabited by normal flora
are the skin, eye, mouth, upper respiratory, GI & urogenital
tracts.
unit-2micro-organisms-.pptx for students
 Beneficial Effects Of The Normal Flora:-
1. The Normal Flora Synthesizes And Excretes Vitamins:-
• For Example, In Humans, Enteric Bacteria Secrete Vitamin K And
Vitamin B12, And Lactic Acid Bacteria Produce Certain B-vitamins.
2. The Normal Flora Prevents Colonization Of Pathogens:-
• By Competing For Attachment Sites. This Is Thought To Be Their
Most Important Beneficial Effect, Which Has Been Demonstrated In
The Oral Cavity, The Intestine, The Skin, And The Vaginal
Epithelium.
3. The Normal Flora May Antagonize (Act-against) Other
Bacteria:- Through The Production Of Substances Which Inhibit Or
Kill Nonindigenous Species. The Intestinal Bacteria Produce A Variety
Of Substances Such As Bacteriocins, Which Inhibit Or Kill Other
Bacteria.
4. The Normal Flora Stimulate The Development Of
Certain Tissues:- The Caecum And Certain Lymphatic
Tissues (Peyer's Patches) In The GI Tract.
5. The Normal Flora Stimulates The Production Of
Natural Antibodies.
 Harmful Effects Of The Normal Flora
1. Bacterial Synergism:- Between A Member Of The
Normal Flora And A Potential Pathogen Helps In
Establishment Of Infection.
• The Normal Flora Supplying A Vitamin Or Some Other
Growth Factor To Other Pathogen Needs In Order To
Grow. This Is Called ”Cross-feeding” Between Microbes.
2. Induction Of A Low Grade Toxemia:- Some Minute
Amounts Of Bacterial Toxins (E.x. Endotoxin) May Be
Found In The Circulation.
3. The Normal Flora May Be Agents Of Disease:-
Members Of The Normal Flora May Cause Endogenous
Disease If They Reach A Site Or Tissue Where They Cannot
Be Restricted Or Tolerated By The Host Defenses.
 Culturing
• “Growing Bacteria Under Artificial Conditions In The
Laboratory Is Called As Culturing.”
OR
• “Growing Of Bacteria Colonies In Artificial Medium
Containing All Suitable Nutrients & Favorable Conditions
Is Called Culturing Of Bacteria.”
• Growth Media Or Culture Media:- To Culture Bacteria
We Provide Them Nutrients That’s Called As ‘Growth
Media’ Or ‘Culture Media.’
• Important Reason Behind Culturing Is Being Utility In
Diagnosing Infectious Disease.
• Glass test tubes & glass based petri dishes are widely used
for cultivating microorganisms.
 Glass Test Tubes & Glass Based Petri Dishes
 Uses Of Culturing Or Culture
Techniques:-
• Isolate Bacteria In Pure Form
• Identify Types Of Bacteria
• Demonstration Of Microbe’s Properties
• Obtain Sufficient Growth For Preparation Of Antigens &
Other Tests.
• Determination Of Antibiotic Sensitivity
• Estimate Viable Counts
 Basic Requirements Of Culture Media:-
• Source Of Energy (Ex. Glucose, Lactose)
• Carbon (CO2)
• Nitrogen (Atmospheric N2)
• Source Of Salts Like Sulphates, Phosphates,
Magnesium, Calcium Etc.
• Optimum pH
• Sterilized Water
 Common Ingredients Of Culture Media:-
• Water
• Agar
• Peptone
• Beef Extract
• Blood
• Serum
• Yeast Extract
• Glucose
 Agar (Just For Knowledge):-
• Agar Or Agar-agar, Is A Jelly-like Substance Consisting
Of Polysaccharides Obtained From The Cell Walls Of Some
Species Of Red Algae, Primarily From “Ogonori” And
"Tengusa“.
• Agar Has Been Used As An Ingredient
In Desserts Throughout Asia And Also Use In Culture
Media For Microbiological Work.
• Agar Can Be Used As A Laxative; An Appetite Suppressant;
A Thickener For Soups; In Fruit Preserves, Ice Cream.
 Broth (Just For Knowledge):-
• It is a liquid medium (soup consistency) containing proteins
and other nutrients for the culture of bacteria.
 Classification Or Types Of Culture Media:-
 On The Basis Of Consistency:-
• Liquid Media :- It Is Used For Biochemical Test & Antibiotic
Sensitivity Motility. (Ex. Meat Extract Broth, Bile Broth)
• Semi Solid Media:- It Is Used To Demonstrate The Motility Of
Bacteria. This Media Looks Very Soft Jelly Like. (Ex. Fluid
Thioglycollate Media)
• Solid Media:- It Is used To Study Colonies Of Individual Bacteria.
It Is Essential For Isolation Of Organism In Pure Form. (Ex.
Nutrient Agar)
 On The Basis Of Oxygen Requirements:-
• Aerobic Media :- Media Which Contains Oxygen Are Called
Aerobic Media.
• Anaerobic Media:- Anaerobic Bacteria Need Special Media For
Growth Because They Need Low Oxygen Content.
 Other Types:-
• Selective Media:- It Is Designed To Suppress The Growth Of
Some Microorganisms While Allowing The Growth Of Others.
• (Ex. Thayer Martin Agar Used To Recover Neisseria
Gonorrhea Contains Vancomycin, Colistin & Nystatin.
• Enrichment Media:- It Is Used To Increase The Relative
Concentration Of Certain Microorganisms.
• (Ex. Selenite F Broth Tetrathionate Broth & Alkaline Peptone
Water Are Used To Recover Pathogens From Fecal
Specimens.)
• Differential Media:- Special Type Of Media Used For
Differentiating Two Varieties Of Bacteria Or Other
Microorganisms.
• (Ex. MacConkey Agar.)
• Basal Or Simple Media:- It Is Basically Simple Media
That Supports Non-fastidious Bacteria.
• (Ex. Peptone Water, Nutrient Broth & Nutrient Agar)
• [Non-Fastidious Bacteria Means:- Bacteria That Do Not
Require Any Special Conditions Or Substances For Their
Growth Are Called Non-fastidious Bacteria.]
 Culture Methods:-
• Streak Plate Culture:- It Is Also Called Surface Plating.
It Is Used For Isolation Of Bacteria In Pure Culture From
Clinical Specimen.
• Lawn Or Carpet Culture:- It Is Prepared By Flooding
The Surface Of Culture Plate With A Liquid Suspension Of
Bacteria By Pipette.
• Stroke Culture:- It Is Made In Tubes Containing Agar
Slope. It Is Used For Producing Pure Growth Of Bacteria
For Slide Agglutination.
• Stab Culture:- It Is Prepared By Puncturing With Charged
Loop, Straight Wire.
• Pour-plate Culture:- This Method Is Used For Counting
The Number Of Living Bacteria Or Groups Of Bacteria In
A Liquid Culture.
• Liquid Culture:- Liquid Culture In Tubes, Bottles Or
Flasks Is Inoculated By Touching With Charged Loop Or
By Pipetting Or By Syringes.
 Staining
• Definition:-“it Is A Technique Which Is Used To Enhance And
Contrast A Biological Specimen At The Microscopic Level By
Using Dyes. This Stains And Dyes Are Used To Highlight The
Specimen At The Microscopic Level To Study It At Higher
Magnification For Histopathological Studies And Diagnostic
Purposes.”
 Application Of Staining / Uses Of Staining
• Used To Increase Visibility Of Microorganisms Being Studied.
• Used To Identify The Shape Of Bacteria.
• Used To Determine The Morphological Features Of
Microorganisms.
• Used To Differentiate & Classify Microorganisms.
 Types Of Stains:-
1. Basic:- Crystal Violet, Gentian Violet, Methylene Blue
2. Acidic:- Eosin, Nigrosin, Indian Ink
3. Neutral:- Giemsa
 Types Of Staining:-
1. Simple Staining
2. Differential Staining
a)Gram’s Stain
b)Albert’s Stain
c)AFB Stain
3. Special Staining
 What Is Smear Preparation Technique?
• The First Step In Most Bacterial Staining
Procedures Is The Preparation Of A Smear.
• In A Smear Preparation, Cells From A Culture Are
Spread In A Thin Film Over A Small Area Of A
Microscope Slide, Dried, And Then Fixed To The
Slide By Heating Or Other Chemical Fixatives.
 What Is Smear Preparation Technique?
 Simple Stain Technique:-
• Staining Can Be Performed With Basic Dyes Such As
Crystal Violet Or Methylene Blue.
• Positively Charged Dyes That Are Attracted To The
Negatively Charged Materials Of Microbial Cytoplasm.
• This Such Procedure Is The Simple Stain Procedure.
unit-2micro-organisms-.pptx for students
 Differential Staining Technique:-
• This Staining Techniques Help To Differentiate Two
Different Types Of Organisms. This Technique Helps To
Separate Bacteria Into Two Groups.
• Differential Staining:-
a)Gram’s Stain
b)Albert’s Stain
c)AFB Stain
 Gram’s Staining Technique:-
• It Was Developed By Christian Gram.
• It Has Widest Application In Bacteriology.
• It Helps To Differentiate Gram+ve & Gram –Ve Bacteria.
• Most Bacteria Can Be Divided Into 2 Groups Based On The
Composition Of Their Cell Wall: Gram Positive & Gram
Negative.
• Gram Positive Cell Walls Stain Blue/Purple With A Gram Stain.
• Gram Negative Cell Walls Stain Pink With A Gram Stain.
• Gram+ve Resist Decolourization & Retain Primary Stain So
Appear Dark Purple/Blue.
• Gram –Ve Get Decolourize So Take The Colour Of Counter Stain
& Appear Pink.
 Differential Stains Of Gram Staining:-
 Steps Of Gram Staining:-
 Steps / Procedure Of Gram Staining:-
• Make A Smear & Fix It.
• Stain The Slide With Crystal Violet For 1 Minute.
• Pour Off The Crystal Violet. (Wash Under Water)
• Flood Slide With Gram’s Iodine For 1 Minute.
• Pour Off The Iodine. (Wash Under Water)
• Add 95% Ethyl Alcohol (Ethanol) For 5-10 Seconds.
• Wash Slide With Water To Remove Ethyl Alcohol. Do Not Delay
This Step.
• Flood Slide With Safranin For 45 Seconds.
• Wash With Water
• Examine Under Microscope.
 List of Gram Positive & Gram Negative
bacteria:-
Gram Positive Gram Negative
Staphylococcus Aureus
Neisseria Meningitis &
Neisseria Gonorrhoeae
Streptococcus Pneumoniae Escherichia Coli (E.Coli)
Pneumococcus Salmonella Typhi
Corynebacterium Diphtheriae Pseudomonas
Clostridium Tetani &
Clostridium Perfringens
Lactobacillus
 Acid Fast Staining Or Ziehl Neelsen’s
Staining:-
• It Is The Differential Staining Techniques Which Was First
Developed By ‘Ziehl’ And Later On Modified By
‘Neelsen’. So This Method Is Also Called Ziehl-neelsen
Staining Techniques.
• The Main Aim Of This Staining Is To Differentiate
Bacteria Into Acid Fast Group And Non-acid Fast Groups.
• This Method Is Used For Those Microorganisms Which
Are Not Staining By Simple Or Gram Staining Method,
Particularly The Member Of Genus Mycobacterium, Are
Resistant And Can Only Be Visualized By Acid-fast
Staining
unit-2micro-organisms-.pptx for students
 Steps / Procedure Of Acid Fast Staining:-
• Prepare A Smear Using Sterile Techniques
• Allow Smear To Air Dry And Then Heat Fix.
• Cover The Smear With Carbol Fuchsin Stain.
• Heat The Stain Until Vapour Just Begins To Rise That Is About
60 C. Do Not Overheat.
ͦ
• Allow The Heated Stain To Remain On The Slide For
5 Minutes.
• Wash Off The Stain With Water.
• Cover The Smear With Acid Alcohol For 15-20 Seconds Or
Until The Smear Is Sufficiently Decolorized, That Is Pale Pink.
• Dry The Slide.
• Examine Under The Microscope.
 List Of Acid Fast & Non Acid Fast
Bacteria:-
Acid Fast Bacteria Non Acid Fast Bacteria
Mycobacterium Tuberculosis Mycoplasma Pneuomaniae
Mycobacterium Leprae Escherichia Coli
Nocardia Brasiliensis Staphylococcus Aureus
Nocardia Cyriacigeorgica Salmonella Typhi
 Albert Staining:-
• It Is Usually Used For ‘Corny Bacterium Diphtheria.’
 Steps / Procedure Of Albert Staining:-
• Prepare A Smear Using Sterile Techniques And
Then Fix It.
• Cover With Albert’s Stain For 5 Min.
• Wash Under Water.
• Cover With Albert’s Iodine For 2 Min.
• Wash Under Water & Dry The Slide.
• Examine Under The Microscope.
unit-2micro-organisms-.pptx for students
unit-2micro-organisms-.pptx for students
 Virus:-
 Virus:-
• “Viruses Are Unicellular, Ultramicroscopic Organisms,
Reproduce Inside Living Cell & Covered By Protein Coat.”
 General Properties Of Virus:-
• They Don’t Have Cellular Organization.
• They Contain Either DNA Or RNA.
• They Multiply By Complex Process.
• They Are Unaffected By Antibiotic.
• They Don’t Have Necessary Enzymes For Protein & Nucleic
Acid Synthesis.
unit-2micro-organisms-.pptx for students
 Size:-
• Largest Virus Measuring 300nm.
• Smallest Virus Measuring 20nm.
 Shape:-
• Viruses Have Different Shape.
• Rabies Virus Has Bullet Shape.
• Pox Virus Has Brick Shape.
• Polio Virus Has Round Shape.
 Structure:- Virus Has Following Parts;
• Central Core Of Nucleic Acid (DNA / RNA).
• Protein Coat Around Core Called Capsid.
• Subunit Of Capsid Is Known As Capsomere.
• Envelope.
 List Of DNA & RNA Viruses:-
DNA Virus RNA Virus
Papilloma Virus Poliovirus
Human Adeno Virus Rhinovirus
Herpes Virus Rabies Virus
Pox Virus Mumps Virus
Hepatitis-B Influenza Virus
Flavi virus
Rubella Virus)
HIV Virus
Hepatitis- A,C,D
 List Of Common Bacterial Diseases:-
Disease Pathogenic Agent Transmission
Botulism
Clostridium
Botulinum
Contaminated Food
Cholera Vibrio Cholerae
Contaminated Food
& Water
Gonorrhoea
Neisseria
Gonorrhoeae
By Sexual Contact
Typhoid Salmonella Typhi
Contaminated Food
& Water
Tetanus Clostridium Tetani
Contaminated
Wound
TB
Mycobacterium
Tuberculosis
Airborne
 List Of Common Viral Diseases:-
Disease Pathogenic Agent Transmission
Rabies Rabies Virus
Bite Of Infected
Animal (Dog)
Genital Herpes Herpes Virus Sexual Transmission
Chicken Pox Varicella Zoster Respiratory Tract
Poliomyelitis Polio Virus Fecal-oral Route
Influenza (Flu) Influenza Virus Respiratory Tract
German Measles Rubella Virus Respiratory Tract
AIDS HIV Sexual Transmission
Common Cold Rhinovirus Respiratory Tract
Dengue
Dengue Virus
(Flavi virus)
Aedes Mosquito
 List Of Common Fungal Diseases:-
 List Of Common Protozoan Diseases:-
Disease Pathogenic Agent
Candidiasis Candida Albicans
Aspergillosis Aspergillus Fumigatus
Sporotrichosis Sporothrix Schenckii
Disease Pathogenic Agent
Amoebiasis Entamoeba histolytica
Malaria Plasmodium species
Giardiasis Giardia lamblia
unit-2micro-organisms-.pptx for students

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unit-2micro-organisms-.pptx for students

  • 1.  Antoni Van Leeuwenhoek Is A ‘Father Of Microbiology’ & ‘Father Of Microscopy.’  Robert Koch (1834-1910) Is Also Known As The 'Father Of Bacteriology‘.  Louis Pasteur Is The Father Of ‘Pasteurization’.  Louis Pasteur Is Also Known As The Father Of ‘Modern Microbiology’.  Elie Metchnikoff (1845-1916) Is Known As The ‘Father Of Phagocytosis’.  Paul Ehrlich (1854-1915) Is Known As The ‘Father Of Chemotherapy’.  Hans Christian Gram (1853-1933) Invented Famous Bacterial Staining Procedure ‘Gram Stain’.
  • 2.  Micro-organism Or Microbe:- “An Organism That Can Be Seen Only Through A Microscope. Microorganisms Include Bacteria, Protozoa, Algae, And Fungi”.  Classification of Micro-organisms:- • Micro-organisms are classified into ten groups: 1. Bacteria 2. Viruses 3. Fungi 4. Protozoa 5. Rickettsiales 6. Algae 7. Moulds 8. Yeast 9. Chlamydia 10. Mycoplasma
  • 4.  Classification of Micro-organisms:- • Classification of microorganism depending on cell type. 1) Eukaryotic micro-organisms 2) Prokaryotic micro-organisms 3) Non cellular micro-organisms  Eukaryotic micro-organisms:- They have complex cellular structure similar to human & animal cell. Eg. Fungi, protozoa, algae  Prokaryotic micro-organisms:- they are simple, unicellular organisms having primitive nuclei & mitochondria. Unicellular also known as single- cell organisms. They are capable to live independent.
  • 5.  Prokaryotic micro-organisms:- they are simple, unicellular organisms having primitive nuclei & mitochondria. Unicellular also known as single- cell organisms. They are capable to live independent. Eg. Bacteria.  Non cellular micro-organisms:- They are without any cell forms called as acellular. Eg. Viruses.
  • 7.  Structure of Micro-Organisms:- • Bacteria are the most well known micro- organisms. • Bacteria are microscopic, prokaryotic, unicellular organisms found everywhere.  Size of the Bacteria:- • Bacteria are very small in size. The unit of measurement of bacteria is called micrometer (mm). • Bacteria generally measure 0.2-1.5mm in diameter & 3.5mm in length.
  • 8.  Morphology Of Bacteria
  • 9.  Structure of Bacteria:-
  • 10.  Morphology Of Bacteria:- Bacteria Is A Prokaryotic, Unicellular Organism. Bacteria Are Found Everywhere, They Are The Most Abundant Living Organisms.  Parts Of Bacteria:- • Bacterial Cell Wall • Capsule • Cytoplasmic Membrane • Cytoplasm • Ribosomes • Mesosomes • Nucleus • Flagella
  • 11. • Bacterial cell wall:- It is complex rigid structure. It gives bacteria its definite shape. It is semi- permeable. The cell wall of gram-positive bacteria is thicker than the gram-negative. • Capsule:- It is a protective covering around the bacterial cell wall. If this capsule is thick it’s called capsule. If this layer is thin it’s called slime layer. Capsule protects the bacteria from antibacterial agents. • Cytoplasmic membrane:- It is also called cell membrane. It is thin & elastic. It is semi-permeable. This membrane carry many enzymes & pigments. • Cytoplasm:- It is viscous watery solution. It contains variety of organic & inorganic solutes & ribosomes.
  • 12. • Ribosomes:- It is 10-20nm in diameter. Function of this is protein synthesis. • Mesosomes:- It is convoluted membranous bodies. • Nucleus:- The bacterial nucleus does not possess nuclear membrane & nucleolus. The genetic information is contained in a single circular, double strand molecule of DNA. • Flagella:- It is long, hollow filaments. They are 10- 20nm in diameter & 3-20mm in length. They are found on large number of bacteria. Flagella helps in motility. • Pili or Fimbriae:- It is hair like structure. They are thinner & shorter than flagella. They allow attachment of bacterial cell to other bacterial cell surfaces.
  • 13. • Flagella Types:- There are 4 types of arrangements of flagella. • 1. Monotrichous:- Single polar flagellum. • 2. Lophotrichous:- Tuft or group of flagella at one pole. • 3. Amphitrichous:- Single flagella at both poles. • 4. Cephalotrichous:- Tuft or group of flagella at both poles. • 5. Peritrichous:- Flagella all around the cell. • 6. Atrichous:- An absence of flagella.
  • 15.  Shape of the Bacteria:- • Individual Bacteria Can Assume One Of 3 Basic Shapes: Spherical (Coccus), Rodlike (Bacillus), Or Curved (Vibrio, Spirillum, Or Spirochete).  Cocci:- They are round or oval in shape. They exist in various patterns. • Types of Cocci:- • Diplococci:- Occurs in pairs, Eg. Gonococci. • Streptococci:- Occurs in chains. • Staphylococci:- Occurs in clusters • Tetrads:- Occurs in Group of 4. • Sarcina (Octads):- Occurs in Group of 8.
  • 17.  Bacilli (Bacillus):- They are rod or stick in shape. It is also called a bacilliform bacterium. They exist in various patterns. •Types of Bacilli:- •Diplobacillus:- Two bacilli arranged side by side with each other. •Streptobacillus:- Occurs in chains. •Coccobacillus:- Oval and similar to Coccus.
  • 19.  Spirilla (Spirillum):- They Are Non-flexible Spiral Forms. They Have One To Three Fixed Curves In Their Body. Eg. Spirillum Volutans.  Vibrio:- They Are Curved Or Comma Shaped Rods. Most Of Them Are Non-pathogenic But Some Are Pathogenic. Eg. Vibrio Cholera  Spirochaetes:- They Are Slender Spiral Filaments. They Are Flexible. They Live In Soil & Stagnant Pools. Eg. Treponema Pallidum
  • 21.  Bacterial Growth:- Bacteria Are Unicellular Organisms That Tend To Reproduce Asexually By The Means Of Binary Fission. • Bacterial Growth Is The Increase In The Number Of Bacterial Cells Rather Than The Increase In Their Cell Size. • The Growth Of These Bacterial Cells Takes Place In An Exponential Manner, i.e., One Cell Divides Into 2, Then 4, Then 8, 16, 32 And So On.
  • 22. • The Time Taken For A Bacterial Cell To Double Is Called ‘Generation Time.’ • The Generation Time Varies Among Different Species Of Bacteria Based On The Environmental Conditions They Grow In. • Clostridium Perfringens Is The Fastest Growing Bacteria That Has A Generation Time Of 10 Minutes. • Escherichia Coli Has A Doubling Time Of 20 Minutes. • Mycobacterium Tuberculosis Is One Of The Slowest Growing Bacteria, Taking About 12 To 16 Hours To Double.
  • 23.  Growth Curve:- • In A Closed System With Enough Nutrients, A Bacteria Shows A Predictable Growth Pattern That Is The Bacterial Growth Curve. • The Bacterial Growth Curve Shows The Preparation, Division, Growth And Death Of The Bacterial Cells. • The Bacterial Growth Progresses In Four Phases Namely – Lag Phase, Log Phase, Stationary Phase And Death Phase. • Bacteria Require Optimum Temperature, Ph, Moisture, Oxygen, Carbon Source, Nitrogen Source And Other Nutrients
  • 25. • 1. Lag Phase:- • The Bacteria Upon Introduction Into The Nutrient Medium Take Some Time To Adapt To The New Environment. • In This Phase, The Bacteria Does Not Reproduce But Prepares Itself For Reproduction. • The Cells Are Active Metabolically And Keep Increasing In Size. • The Cells Synthesise RNA, Growth Factors And Other Molecules Required For Cell Division.
  • 26. • 2. Log Phase:- • Soon after the lag phase, i.e., the preparation phase, the bacterial cells enter the log phase. • This phase is also known as the exponential phase. • This phase is marked by the doubling of the bacterial cells. The cell number increases. • The log phase continues until there is depletion of nutrients in the setup. • The stage also comes to a stop if toxic substances start to accumulate, resulting in a slower growth rate. • The cells are the healthiest at this stage and researchers prefer to use bacteria from this stage for their experimental processes.
  • 27. • 3. Stationary Phase:- • In The Stationary Phase, The Rate Of Growth Of The Cells Becomes Equal To Its Rate Of Death. • The Rate Of Growth Of The Bacterial Cells Is Limited By The Accumulation Of Toxic Compounds And Also Depletion Of Nutrients In The Media. • The Cell Population Remains Constant At This Stage. • 4. Death Phase:- • This Is The Last Phase Of The Bacterial Growth. • At This Stage, The Rate Of Death Is Greater Than The Rate Of Formation Of New Cells. • Lack Of Nutrients, Physical Conditions Or Other Injuries To The Cell Leads To Death Of The Cells.
  • 28.  Factors Influencing Growth Of Bacteria Or Microbes:- 1. Nutrition 2. Moisture 3. Temperature 4. pH 5. Oxygen 6. Carbon Dioxide 7. Light & Other Radiations 8. Osmotic Effect 9. Mechanical & Sonic Stresses
  • 29. 1. Nutrition:- The Growth Of Microbe Depend On Adequate Supply Of Suitable Nutrients Like Carbon, Nitrogen, Phosphates, Ions & Metals. 2. Moisture:- It Is Very Essential For The Growth Of Bacteria Because 80% Of The Body Of Bacteria Is Made Up Of Water. Moist Places Are More Viable Media For Growth Of Bacteria. 3. Temperature:- Bacteria Requires Specific Temp. Range For Their Growth. Bacteria Can Be Killed By Very High Temp. & Very Low Temp. Favorable Range Is 20 - 40 C. ͦ ͦ
  • 30. 4. pH:- Some Bacteria Grow In Neutral Media, Some Grow In Alkaline Media & Some Grow In Acidic Media. • Acidophiles:- Organisms grow at pH between 0.5-6. • Neutrophiles:- Organisms grow best at pH 7. • Alkaliphiles:- Organisms grow best under at pH above 7. 5. Oxygen:- The Bacteria Which Can Grow Only In The Presence Of Oxygen Are Known As ‘Aerobic Bacteria’. This Bacteria Requires Oxygen For Life & Reproduction. • Some Bacteria Can Grow In The Absence Of Oxygen Are Known As ‘Anaerobic Bacteria’. This Bacteria Does Not Requires Oxygen For Life & Reproduction. 6. Carbon Dioxide:- Some Bacteria Requires Extra CO2 In The Air For Their Growth & Reproduction.
  • 31. 7. Light & Other Radiations:- Bacteria Grow In Dark Places. Bacteria Can Be Destroyed Or Killed By UV Rays, Ionizing Radiations, Infrared Rays & Other Radiations. 8. Osmotic Effect:- Osmotic Pressure Changes Can Effect The Growth Of Bacteria. Sudden Changes In Osmotic Pressure Can Cause Destruction Of Bacteria. 9. Mechanical & Sonic Stresses:- Bacteria Cell Wall Can Be Ruptured & Bacteria Can Be Killed By Vigorous Shaking & Ultrasonic Vibrations.
  • 32.  Normal Flora:- • The Normal Flora Are Micro-organisms Which Are Found In Or On Our Body Without Causing Any Disease. • Normal Flora Arranged In Two Groups: 1. Resident Flora:- It Is Consist Of Permanent Microorganisms Which Are Regularly Found In A Given Area. 2. The Transient Flora:- It Is Consist Of Pathogenic Or Non-pathogenic Organisms That Stay On The Skin Or Mucus Membranes For Hours, Day Or Week. They Do Not Stay Permanently On The Surface. • The most common sites of body inhabited by normal flora are the skin, eye, mouth, upper respiratory, GI & urogenital tracts.
  • 34.  Beneficial Effects Of The Normal Flora:- 1. The Normal Flora Synthesizes And Excretes Vitamins:- • For Example, In Humans, Enteric Bacteria Secrete Vitamin K And Vitamin B12, And Lactic Acid Bacteria Produce Certain B-vitamins. 2. The Normal Flora Prevents Colonization Of Pathogens:- • By Competing For Attachment Sites. This Is Thought To Be Their Most Important Beneficial Effect, Which Has Been Demonstrated In The Oral Cavity, The Intestine, The Skin, And The Vaginal Epithelium. 3. The Normal Flora May Antagonize (Act-against) Other Bacteria:- Through The Production Of Substances Which Inhibit Or Kill Nonindigenous Species. The Intestinal Bacteria Produce A Variety Of Substances Such As Bacteriocins, Which Inhibit Or Kill Other Bacteria.
  • 35. 4. The Normal Flora Stimulate The Development Of Certain Tissues:- The Caecum And Certain Lymphatic Tissues (Peyer's Patches) In The GI Tract. 5. The Normal Flora Stimulates The Production Of Natural Antibodies.  Harmful Effects Of The Normal Flora 1. Bacterial Synergism:- Between A Member Of The Normal Flora And A Potential Pathogen Helps In Establishment Of Infection. • The Normal Flora Supplying A Vitamin Or Some Other Growth Factor To Other Pathogen Needs In Order To Grow. This Is Called ”Cross-feeding” Between Microbes.
  • 36. 2. Induction Of A Low Grade Toxemia:- Some Minute Amounts Of Bacterial Toxins (E.x. Endotoxin) May Be Found In The Circulation. 3. The Normal Flora May Be Agents Of Disease:- Members Of The Normal Flora May Cause Endogenous Disease If They Reach A Site Or Tissue Where They Cannot Be Restricted Or Tolerated By The Host Defenses.
  • 37.  Culturing • “Growing Bacteria Under Artificial Conditions In The Laboratory Is Called As Culturing.” OR • “Growing Of Bacteria Colonies In Artificial Medium Containing All Suitable Nutrients & Favorable Conditions Is Called Culturing Of Bacteria.” • Growth Media Or Culture Media:- To Culture Bacteria We Provide Them Nutrients That’s Called As ‘Growth Media’ Or ‘Culture Media.’ • Important Reason Behind Culturing Is Being Utility In Diagnosing Infectious Disease. • Glass test tubes & glass based petri dishes are widely used for cultivating microorganisms.
  • 38.  Glass Test Tubes & Glass Based Petri Dishes
  • 39.  Uses Of Culturing Or Culture Techniques:- • Isolate Bacteria In Pure Form • Identify Types Of Bacteria • Demonstration Of Microbe’s Properties • Obtain Sufficient Growth For Preparation Of Antigens & Other Tests. • Determination Of Antibiotic Sensitivity • Estimate Viable Counts
  • 40.  Basic Requirements Of Culture Media:- • Source Of Energy (Ex. Glucose, Lactose) • Carbon (CO2) • Nitrogen (Atmospheric N2) • Source Of Salts Like Sulphates, Phosphates, Magnesium, Calcium Etc. • Optimum pH • Sterilized Water
  • 41.  Common Ingredients Of Culture Media:- • Water • Agar • Peptone • Beef Extract • Blood • Serum • Yeast Extract • Glucose
  • 42.  Agar (Just For Knowledge):- • Agar Or Agar-agar, Is A Jelly-like Substance Consisting Of Polysaccharides Obtained From The Cell Walls Of Some Species Of Red Algae, Primarily From “Ogonori” And "Tengusa“. • Agar Has Been Used As An Ingredient In Desserts Throughout Asia And Also Use In Culture Media For Microbiological Work. • Agar Can Be Used As A Laxative; An Appetite Suppressant; A Thickener For Soups; In Fruit Preserves, Ice Cream.  Broth (Just For Knowledge):- • It is a liquid medium (soup consistency) containing proteins and other nutrients for the culture of bacteria.
  • 43.  Classification Or Types Of Culture Media:-  On The Basis Of Consistency:- • Liquid Media :- It Is Used For Biochemical Test & Antibiotic Sensitivity Motility. (Ex. Meat Extract Broth, Bile Broth) • Semi Solid Media:- It Is Used To Demonstrate The Motility Of Bacteria. This Media Looks Very Soft Jelly Like. (Ex. Fluid Thioglycollate Media) • Solid Media:- It Is used To Study Colonies Of Individual Bacteria. It Is Essential For Isolation Of Organism In Pure Form. (Ex. Nutrient Agar)  On The Basis Of Oxygen Requirements:- • Aerobic Media :- Media Which Contains Oxygen Are Called Aerobic Media. • Anaerobic Media:- Anaerobic Bacteria Need Special Media For Growth Because They Need Low Oxygen Content.
  • 44.  Other Types:- • Selective Media:- It Is Designed To Suppress The Growth Of Some Microorganisms While Allowing The Growth Of Others. • (Ex. Thayer Martin Agar Used To Recover Neisseria Gonorrhea Contains Vancomycin, Colistin & Nystatin. • Enrichment Media:- It Is Used To Increase The Relative Concentration Of Certain Microorganisms. • (Ex. Selenite F Broth Tetrathionate Broth & Alkaline Peptone Water Are Used To Recover Pathogens From Fecal Specimens.) • Differential Media:- Special Type Of Media Used For Differentiating Two Varieties Of Bacteria Or Other Microorganisms. • (Ex. MacConkey Agar.)
  • 45. • Basal Or Simple Media:- It Is Basically Simple Media That Supports Non-fastidious Bacteria. • (Ex. Peptone Water, Nutrient Broth & Nutrient Agar) • [Non-Fastidious Bacteria Means:- Bacteria That Do Not Require Any Special Conditions Or Substances For Their Growth Are Called Non-fastidious Bacteria.]
  • 46.  Culture Methods:- • Streak Plate Culture:- It Is Also Called Surface Plating. It Is Used For Isolation Of Bacteria In Pure Culture From Clinical Specimen.
  • 47. • Lawn Or Carpet Culture:- It Is Prepared By Flooding The Surface Of Culture Plate With A Liquid Suspension Of Bacteria By Pipette.
  • 48. • Stroke Culture:- It Is Made In Tubes Containing Agar Slope. It Is Used For Producing Pure Growth Of Bacteria For Slide Agglutination.
  • 49. • Stab Culture:- It Is Prepared By Puncturing With Charged Loop, Straight Wire.
  • 50. • Pour-plate Culture:- This Method Is Used For Counting The Number Of Living Bacteria Or Groups Of Bacteria In A Liquid Culture.
  • 51. • Liquid Culture:- Liquid Culture In Tubes, Bottles Or Flasks Is Inoculated By Touching With Charged Loop Or By Pipetting Or By Syringes.
  • 52.  Staining • Definition:-“it Is A Technique Which Is Used To Enhance And Contrast A Biological Specimen At The Microscopic Level By Using Dyes. This Stains And Dyes Are Used To Highlight The Specimen At The Microscopic Level To Study It At Higher Magnification For Histopathological Studies And Diagnostic Purposes.”  Application Of Staining / Uses Of Staining • Used To Increase Visibility Of Microorganisms Being Studied. • Used To Identify The Shape Of Bacteria. • Used To Determine The Morphological Features Of Microorganisms. • Used To Differentiate & Classify Microorganisms.
  • 53.  Types Of Stains:- 1. Basic:- Crystal Violet, Gentian Violet, Methylene Blue 2. Acidic:- Eosin, Nigrosin, Indian Ink 3. Neutral:- Giemsa  Types Of Staining:- 1. Simple Staining 2. Differential Staining a)Gram’s Stain b)Albert’s Stain c)AFB Stain 3. Special Staining
  • 54.  What Is Smear Preparation Technique? • The First Step In Most Bacterial Staining Procedures Is The Preparation Of A Smear. • In A Smear Preparation, Cells From A Culture Are Spread In A Thin Film Over A Small Area Of A Microscope Slide, Dried, And Then Fixed To The Slide By Heating Or Other Chemical Fixatives.
  • 55.  What Is Smear Preparation Technique?
  • 56.  Simple Stain Technique:- • Staining Can Be Performed With Basic Dyes Such As Crystal Violet Or Methylene Blue. • Positively Charged Dyes That Are Attracted To The Negatively Charged Materials Of Microbial Cytoplasm. • This Such Procedure Is The Simple Stain Procedure.
  • 58.  Differential Staining Technique:- • This Staining Techniques Help To Differentiate Two Different Types Of Organisms. This Technique Helps To Separate Bacteria Into Two Groups. • Differential Staining:- a)Gram’s Stain b)Albert’s Stain c)AFB Stain
  • 59.  Gram’s Staining Technique:- • It Was Developed By Christian Gram. • It Has Widest Application In Bacteriology. • It Helps To Differentiate Gram+ve & Gram –Ve Bacteria. • Most Bacteria Can Be Divided Into 2 Groups Based On The Composition Of Their Cell Wall: Gram Positive & Gram Negative. • Gram Positive Cell Walls Stain Blue/Purple With A Gram Stain. • Gram Negative Cell Walls Stain Pink With A Gram Stain. • Gram+ve Resist Decolourization & Retain Primary Stain So Appear Dark Purple/Blue. • Gram –Ve Get Decolourize So Take The Colour Of Counter Stain & Appear Pink.
  • 60.  Differential Stains Of Gram Staining:-
  • 61.  Steps Of Gram Staining:-
  • 62.  Steps / Procedure Of Gram Staining:- • Make A Smear & Fix It. • Stain The Slide With Crystal Violet For 1 Minute. • Pour Off The Crystal Violet. (Wash Under Water) • Flood Slide With Gram’s Iodine For 1 Minute. • Pour Off The Iodine. (Wash Under Water) • Add 95% Ethyl Alcohol (Ethanol) For 5-10 Seconds. • Wash Slide With Water To Remove Ethyl Alcohol. Do Not Delay This Step. • Flood Slide With Safranin For 45 Seconds. • Wash With Water • Examine Under Microscope.
  • 63.  List of Gram Positive & Gram Negative bacteria:- Gram Positive Gram Negative Staphylococcus Aureus Neisseria Meningitis & Neisseria Gonorrhoeae Streptococcus Pneumoniae Escherichia Coli (E.Coli) Pneumococcus Salmonella Typhi Corynebacterium Diphtheriae Pseudomonas Clostridium Tetani & Clostridium Perfringens Lactobacillus
  • 64.  Acid Fast Staining Or Ziehl Neelsen’s Staining:- • It Is The Differential Staining Techniques Which Was First Developed By ‘Ziehl’ And Later On Modified By ‘Neelsen’. So This Method Is Also Called Ziehl-neelsen Staining Techniques. • The Main Aim Of This Staining Is To Differentiate Bacteria Into Acid Fast Group And Non-acid Fast Groups. • This Method Is Used For Those Microorganisms Which Are Not Staining By Simple Or Gram Staining Method, Particularly The Member Of Genus Mycobacterium, Are Resistant And Can Only Be Visualized By Acid-fast Staining
  • 66.  Steps / Procedure Of Acid Fast Staining:- • Prepare A Smear Using Sterile Techniques • Allow Smear To Air Dry And Then Heat Fix. • Cover The Smear With Carbol Fuchsin Stain. • Heat The Stain Until Vapour Just Begins To Rise That Is About 60 C. Do Not Overheat. ͦ • Allow The Heated Stain To Remain On The Slide For 5 Minutes. • Wash Off The Stain With Water. • Cover The Smear With Acid Alcohol For 15-20 Seconds Or Until The Smear Is Sufficiently Decolorized, That Is Pale Pink. • Dry The Slide. • Examine Under The Microscope.
  • 67.  List Of Acid Fast & Non Acid Fast Bacteria:- Acid Fast Bacteria Non Acid Fast Bacteria Mycobacterium Tuberculosis Mycoplasma Pneuomaniae Mycobacterium Leprae Escherichia Coli Nocardia Brasiliensis Staphylococcus Aureus Nocardia Cyriacigeorgica Salmonella Typhi
  • 68.  Albert Staining:- • It Is Usually Used For ‘Corny Bacterium Diphtheria.’
  • 69.  Steps / Procedure Of Albert Staining:- • Prepare A Smear Using Sterile Techniques And Then Fix It. • Cover With Albert’s Stain For 5 Min. • Wash Under Water. • Cover With Albert’s Iodine For 2 Min. • Wash Under Water & Dry The Slide. • Examine Under The Microscope.
  • 73.  Virus:- • “Viruses Are Unicellular, Ultramicroscopic Organisms, Reproduce Inside Living Cell & Covered By Protein Coat.”  General Properties Of Virus:- • They Don’t Have Cellular Organization. • They Contain Either DNA Or RNA. • They Multiply By Complex Process. • They Are Unaffected By Antibiotic. • They Don’t Have Necessary Enzymes For Protein & Nucleic Acid Synthesis.
  • 75.  Size:- • Largest Virus Measuring 300nm. • Smallest Virus Measuring 20nm.  Shape:- • Viruses Have Different Shape. • Rabies Virus Has Bullet Shape. • Pox Virus Has Brick Shape. • Polio Virus Has Round Shape.  Structure:- Virus Has Following Parts; • Central Core Of Nucleic Acid (DNA / RNA). • Protein Coat Around Core Called Capsid. • Subunit Of Capsid Is Known As Capsomere. • Envelope.
  • 76.  List Of DNA & RNA Viruses:- DNA Virus RNA Virus Papilloma Virus Poliovirus Human Adeno Virus Rhinovirus Herpes Virus Rabies Virus Pox Virus Mumps Virus Hepatitis-B Influenza Virus Flavi virus Rubella Virus) HIV Virus Hepatitis- A,C,D
  • 77.  List Of Common Bacterial Diseases:- Disease Pathogenic Agent Transmission Botulism Clostridium Botulinum Contaminated Food Cholera Vibrio Cholerae Contaminated Food & Water Gonorrhoea Neisseria Gonorrhoeae By Sexual Contact Typhoid Salmonella Typhi Contaminated Food & Water Tetanus Clostridium Tetani Contaminated Wound TB Mycobacterium Tuberculosis Airborne
  • 78.  List Of Common Viral Diseases:- Disease Pathogenic Agent Transmission Rabies Rabies Virus Bite Of Infected Animal (Dog) Genital Herpes Herpes Virus Sexual Transmission Chicken Pox Varicella Zoster Respiratory Tract Poliomyelitis Polio Virus Fecal-oral Route Influenza (Flu) Influenza Virus Respiratory Tract German Measles Rubella Virus Respiratory Tract AIDS HIV Sexual Transmission Common Cold Rhinovirus Respiratory Tract Dengue Dengue Virus (Flavi virus) Aedes Mosquito
  • 79.  List Of Common Fungal Diseases:-  List Of Common Protozoan Diseases:- Disease Pathogenic Agent Candidiasis Candida Albicans Aspergillosis Aspergillus Fumigatus Sporotrichosis Sporothrix Schenckii Disease Pathogenic Agent Amoebiasis Entamoeba histolytica Malaria Plasmodium species Giardiasis Giardia lamblia