BACTERIA
Staining 1 Basic Stains
Process of colouring the objec
t ( tissue or organisms)
Stains : these are salts used fo
r staining.
Staining Component is Basic
e.g Methylene Blue
Crystal violet Basic fuchsin
Acidic Stains
Negrocin Indian ink
Eosin
2. Synthetic Stains
Most of stains are ....
e.g Methylene Blue
C.violet Basic Fuchsin ....
Natural Hematoxyline
 Staining 2 Progressive
1 Simple Staining Tissue staining in which different
( Direct Staining ) stains are used witout decolouriz
er( alcohol)
Only 1stain is used
e.g Hematoxyline & Eosin
e.g Methylene Blue
( H & E )Saininig
Compound Staining
Regressive Staining
( indirect Staining )
Decolourizer is used
> 1 Stains are used or
e.g Gram's Staining
Mordant is applied
Z& N
e.g Gram's Staining
Z& N Staining
Obligate aerobics Obligate anaerobs
Nocardia Peptococcus
Pseudomonas peptostreptococcus
Mycobacterium S.Saprophyticus
Facultatative anaerobs Gram _ ve cocci
Most streps & Veillonella
S.aureus S.Epidermis G+ ve Rods
Bacillus anthracis & Cereus Clostridiae
Neisseriae Propionibacterium
All enterobacteriacae HCV Lactobacilus
C.DiptheriaeL.monocytogen Actinomyces
Pasrurella & Yersenia Bifidobacterium
Gram _ Rods
Virulance Factors Bacteri
a
Special Biochemical
characterstiscs Tests

Growth
 S. Pyogen
• A. CELL WALL
1. C Carcbohydrate B ENZYMES
2 . M PROTEIN 1 Streptokinase
2. StreptoDnase
B. EXOTOXINS 3. Hylurinidase
1. Erythrogenic 4 Strepylolysin O& S
( scarlet fever)
2. EXOTOXIN A GROWTH
3.EXOTOXIN B Crystal violet Blood Agar
Classification of Streptoco
cci
On 4 Basis
1 Lancified ( Group A B C D E ...)
2 Hemolysis ( Beta alpha gamma )
3 Biochemical
(1 Hipurate Hydrolsis 2 Growth in Hypertonic Saline
3 Bile Solubility 4 Mannitol & Lactose)
4 Dug Sensitivity
1 Bacitracin 2 Optochin
Biochemical Tests
Beta hemolytic

Bacitracin Sensitive Resitant &

Hipurate Hydrolysis
CAMP +

S .pyogen S.agalctiae(neomycin Blood aga

 Hypertonic Saline (6.5%) Bile Soluble &
Growth Optochin Sensitive
+ _ (alpha hemolytic)
+

Ent.Faecalis Ent.bovis
_
LITMUS milk Reduction S.pneumoniae
Test
+
Viridiants
Mannitol +ve
Lactose +ve
Streptococci
1 All grow on Blood agar
2 All R Catalase _
3 Only S.Pyogen is Bacitracin +
4 Only S.agalactiae is CAMP +
5 Only S.pneumoniae Bile & Optochin +
6 Only Enterococci Lactose &Mannitol +
7 Viridians All _
 • EXOTOXINS
S. AUREUS1. Toxic Shock Syndrone Tox
in
( TSST )
• A. CELL WALL 2 Exfoliative
• Protein A 3 Leukocidins(alpha & PV)
• Teichoic Acid 4. Exotoxin alpha
(Hemolysis
• B.ENzyMES 5 Exotoxin beta
• 1. Staphylokinase • ( sphingomylin )
6. Entertoxin (superantige
• 2. Hylurunidase n)
• 3. Catalse Heat Stable / 6 types
• 4. Coagulase Grows on DNAs
e agar & Mannitol salt agar
• 5. DNASe
• 6. Lipase(C.Botulinium)
Biochemical Tests
All R catalase + ve
Only S.aureus is Coagulase +ve

S.Epidermis Coagulase -ve

Novobiocin +ve
& S.Saprophyticus Coagulase -ve
Novobiocin -ve
N.GONORHEA NMENINGITID
IS
• 1. PILLI(>100) • 1. PILLI
• 2. PROTEINS • 2. Capsule(13 sero)
• Por • 3. PROTEINS ( 1- 5)
• Opa • 123 = por protein
• Rmp • 5 = opa
• 3. Ig A protease • 4. Ig A protease
• 4. LOG • 5. LOG
• ( Endotoxin ) • ( Endotoxin )
 Similarties B/W Gonorrhea &
Meningitis
10 Catalse +ve
1Gm -ve Diplococc
11 Oxidase +ve
2 Facultative anaerob 12 Glucose +ve
3 Non Motile 13 Lactose -ve
4 Chocolate agar 14 Fructose -ve
5 10% Co2 (Capnophil 15 Sucrose-ve
lic) 16 DNAse -ve
6Ig A protease 17 Blood agar -ve
7 LOG Endotoxin 18 Nutient Agar -ve
8 Human Habitat 19 VCTN Media +ve
20 Pilli
 Difference
s
1 Non Capsulted 1 Capsulted
2 Sexually Transmission 2 Respiratory droplets
3 > 100 Serotypes (Pilli) 3 > 13 Serotypes
4 Gonorhea 4 Meningitis
5 Neonatal Opthalmia 5 > 2yrs Children Mening
6 Genital & joints 6 No genitalia
7 Urine Sample 7 CSF Sample
8 Rarely BetaLactamase 8 Beta Lactamase
9 No Quelling Reaction 9 Quelling Reaction +ve
10 No Vaccine 10 Vaccines available
11 Maltose - ve 11 Maltose +ve
 H. INFLUENZAE
• 1. PILLI Nonmotile
Non Hemolytic
D/D H.parahemolyticus &
• 2. Capsule
H. Hemolytic R Hemolytic
• ( 6 types polysaccharide)
Glucose
Polyribitol phosphate
Ribose +
• 3. Ig A protease Xylose
• 4. LPS
Indole V
( Endotoxin )
Urease

Lactose -
 B.Pertusis
• 1. Pilli
2 Filamentous Hemagluti
n ( FHA) 1 Catalse vM
3 Capsule 2 Oxidase +
4 EXOTOXINS 3 Urease _
• 1. Pertusis Toxin
• 2. Tracheal Cytotoxin 4 Growth on
• 3. Adenyl Cyclase Blood agar
5 ENDOTOXIN Choclate

Aerobic Mackonky _
S-S Agar
Non Fermentator
Non motile
Legionella
• 1. Flagellum ( H Ag) 1 Tyrosine
• 2. Capsule 2 Gelatin
• 3. Enzymes 3 Hippurate +
• Protease lipase phosp 4 Beta Lactamase
hatase RNAse
• DNAse 5 Oxidase V

4. Endotoxin
B.antrhacis B. Ce
reus
1. Edema Factor Heat Labile enterotoxin
2. Lethal Factor ( Diarrhea type )
3. Protective Ag Heat Stable enterotoxin
( vomitting Type)
Glucose +
Sucrose +
Maltose +
Lecithinase + _
C. Tetanus C. Bot
ulinum
• Tetanospasmin Botulinum Toxin
Lipase +ve
Gelatin Hydrolysis +ve Gelatin Hydrol
DNAse V DNAse
Lactose
Lactose
Lecithinase _
Lecithinase
Lipase Mannitol
Mannitol Proteolytic activity CDE
Proteolytic activity ABF +
Botulinum Tetanus Similarit
ies
1 G+ve 9 Gelatin Hydrolysis +
2 Anaerobic 10 Lactose
3 Paralysis 11 Lecithinase _
4 Most Toxic Exotoxin 12 Mannitol
5 No endotoxin
6 Anaerobic Growth
7 Bile agar Growth
8 Can spread by Wound
Tetanus VS Botul
inum
1 Terminal Spores 1
( Drumstick )
2 Spastic Paralysis 2 Flaccid paralysis
3 Spreads only by wo 3 can also spread by food
und ingestion
4 Lipase _ 4 lipase +
C.Perfringes C.
Difficle • Toxin A
•alpha Toxin
• Lecithinase Enteotoxin
Enterotoxin • Toxin B
Like S. Aureus • Lysis Cell membrane

Gelatin Hydrolysis +ve

• Mannitol +
DNAse +
Gelatin Hydrolysis _
Lactose +
DNAse _
Lecithinase +
Mannitol _ Lactose _
Lipase _ Lecithinase _
Proteolytic activity Lipase _
Corrynebacterium Diptherae

• Dipthera Toxin Glucose

• ( polypeptide) Maltose +
• Fragment A
ADPRibosylation Nitrate reduction V
Nonhemolytic
Urease
• Fragment B
Alk: phosphatase _
Binding site
Sucrose
H2 S _
Listeria Monocytogen

1 listeriolysin O Catalase
Glucose
2. Internalin
Maltose +
Esculin
3. Hemolysin
( Beta hemolysis )
Umbrella Growth in Mannitol _
Stab culture H2 S _
EXOTOXINS
Most powerful Toxins
1 Botulinum
2 Diptherea
3 Shigella
4 Tetanus
They all R neurotoxins as
target organ is distant fro
m primary site
Superantigens
1. TSST by S.A + SuperAg activate T Cells
2 Erythrogenic Toxin S.py release Cytokine
gen IFN gamma Macrophage
( Scarlet fever ) s IL 1 & TNF stimulate en
teronervous System
3 .Enterotoxins of
Vomitting
4 S.A & C. Perfringes
5 Lecithinase by C.Perfrin
ges
TELEE
Diseases By Superantigens
Scarlet fever Kawasaki's Disease
Toxic Shock Syndrome Nasal polyp
Diarhea DM
( S.aureus +C.Perfering
es)
RA
Eczema
Psoriasis
 Diarrhea Causing Bacteria
Watery Bloody+ Fever
B.anthracis
B.Cereus E.Coli ( Shiga toxin )
S. A = entereotoxin EPEC EHEC EIEC
C.perfringes ===== Shigella
C. Difficle enrter Campylobacter
otoxin Y.entericolitis
E.Coli entero TB Intestine
Cholera ======
S.Typhimumi
Parahemolyticus ( F)
S.Typhi (F)
BEST CTY
 Pseudomonas
• 1. Flagella ( H Ag ) • 8 ENzyMes
• 2. Pilli • 1. Hemolysin ??
• 3. Capsule • 2. Elastase
• 4. Pyocinin • 3. Collagenase
• 5. Endotoxin
• 4 DNAse
• 6 Exotoxin A
• 5 Fibrinolysin
• ( like Diptherea)
• 7. Exotoxin S
Aerobic
• ( type lll secretory Strai Non Fermentator
n)
Oxidase +
Biochemical Tests
. .
(Pseudomons)
1 Oxidase 1 Maltose
2 pyoverdin 2 Mannitol V
3 Arginine + 3 Urea
4 Acetamide

1 Lactose
2 Lysine _
3 Esculine
E.Coli
• 1. Flagella ( H Ag ) • 7 EXOTOXINS (enterot
• 2.pilli oxin)
• 3.Capsule ( k Ag ) 1. Heat labil ( LT)
• 4. Endotoxin ( O Ag ) Enterotoxin = Watery
• 5 Adhesins 2 Heat Stable (ST)
• 6 Siderophores 3. Shiga Toxin = Bloody
( Verotoxin) 0157: H7
By Bacteriophage
Biochemical E.Coli
Lactose TSI
Mannitol + Butt & Slant Yellow
H2S _
Glucose
Gas +
Sucrose V
MIU
Oxidase _ (motility indole urea)
Citrate Motile
Indole + ( EHEC _ )
Urea _
EXOTOXINS
• ADP Ribosylation Diptheria+Pseudomons
1 Botulinum Toxin (8) Elongation Factor ( EF) inhi
bit Protein Synthesis
2 B. Pertusis Toxin
Cholera + E.Coli
3 Cholera Toxin
ADP Ribosylate G5protein
4 Diptherea activates
5 E.Coli ( entero LT ) Adenylate Cyclase CAmp
6 Tetanus Diarrhea
7 pseudomonas( Exo A) Pertusis Gi protein
VIBRIACEAE
H PYLORI • Vibrio Cholerae
1 H Ag ( Flagella) 1 H Ag ( Heat labile)
2 Urease 2 Endotoxin
3 LPS /Endotox 3 Capsule
Compylobacter ( some species)
1 H Ag 4 Mucin & Fimbriae
2Cytotoxin 5 Coleragen
3 Endotoxin E.Coli ( LT)like Diarhea
 V. Cholerae
Biochemical Glucose
Sucrose (_ V.Para)
H.pylori Mannitol +
Oxidase
Compylobacter
Indole
Citrate V
Urea
Lactose _
H2S
Gas
 Biochemical enterobacteria
& related Sucrose +
1 All R Glucose +
Klebsiella
2 All R Mannitol + exc
ept Proteus Morganella providentia
& Pseudomons Y.entericolitica
3 All R Oxidase _ V. Cholerae
But 4 Indole +
Pseudomons E.coli
vibrio (Oxidase) Morganella
Compylobacter + P. Vulgaris (P.mirables)
Vibrios
M.Tuberculosis
1. Mycolic Acid 5 Phthiocerol Dimycoce
( Carb C 78 - C90) ocate
2 Cord Factor
( trehelose Dimycolate)
3 Proteins
( PPD skin test )
4 Wax D
Properties
Catalase 68% _ Non motile
Lipase
Noncapsulated
Nitrate +
NAP inhibition No endotoxin
Amidase No Exotoxin
Peroxidase
Neutral Red test
Pyrrazinamidase
1. All R
Zoonotics
Facultatative intrac
ellular parasites except past 6 All R catalase + but
urella
Francisella??
2 All R Capsulted except Br
ucella 7 Brucella is all +
2 All R nonmotile except Y.e Catalase Oxidase urease
ntericolitis
3. All R Coccobacillay 8 Only Yersenia grows on
4 All R Bipolar Staining Mackconkey
+ 9 oxidase + Brucelle & Fra
5 All present wit tender ncisella
lymphadenopathy except
Pasturella (cellulitis)
Zoonotics
Brucella
1 Glucose Francisella
2 Maltose + Capsule
3 Glycerol Endotoxin
4 Oxidase
5 Urease
6 Catalase
7 Basic Fuchsin Sensitivity

Growth on MacConkey
V
 Pasturella 1 Oxidase
Aerobic 2 Catalase
Cellulitis 3 Mannitol
Capsule 4 Indole +
Nonmotile 5 Galactose
Nonhemolytic 1 Maltose
2 urease
3 V-P
Yersenia
Yersenia pestis 1 Catalase
Capsule 2 Glucose
F-1 Ag ( fraction-1) 3 Maltose
V-W Ag 4 Mannitol
Bacteriocin ((Pesticin ) 5 Oxidase
Endotoxin 6 Nonlacose ( white colo
Exotoxin nies on McConky)
Cardiotoxic for animals
Catalase + Oxidase +
All Staphylococci Listeria
Neisseriae Neisseriae
All enterobactereca Bordetella
Listeria Monocytogen Vibrio Compylobacter
Compylobacter Pseudomons
All Zoonotics except Brucella Pasturella
Francisella Urease +
Catalase _ H.pylori
All Streptococci Proteus Providentia
All Clostridiae Morganella
Coagulase + Brucella
S. Aureus Y.pestis U.urealyticum
Motile Non Motil
e B.anthracis All Cocci
B.Cereus C.perfringes
All Clostridiae except C.Diptheriae Actinomyce
Listeria Monocytogen s
Legionella Nocardia
H.influenzae
All enterobacteriacae exce Bordetella
pt
H.pylori Klebsiella & Shigella
Compylobacter ( HCV ) Bacteroids
Vibrio Cholerae Chlamydia Ricketsia
Y. enterocolitis All zoonotics except
Pseudomonas
Mycobacterium
All Spirochetes
Mycoplasma
Capsulted Non Caps
ulted
S.pneumoniae N.gonorheae
S.pyogen ( Some ) All G+ Bacilli R noncapsultedexce
S.aurues ( some) pt anthracis
N.meningitidis Proteus
B.antracis Chlamydia
All Respiratory G_ ve Ricketsia
HBL
Spirochetes
All enterobacteriacae
Mycobacterium
except Proteus
Mycoplasma
V.cholerae
All zoonotics except Bruce
lla
Pseudomonas
 Vaccines
Live organisms Capsular polysach:
TB H.influenzae
Typhoid (+capsular) N.meningitidis
Tularemia S.pneumoniae
Killed Organisms Typhoid( live)
B.pertusis(+ acellular) Toxoid
V.cholerae Diptherea
R.powazekii ( Typhus) Tetanus
Purified protein(partially)
Coxiella burnetii ( Q fever)
B.anthracis
Y.pestis (plaque)
B.pertusis
No Vaccine for enterobacteria (ex
cept No vaccines for all Cocci exce
pt S.pneumoniae &
S.typhoid & V.cholerae