Bacteria GNB

A. ESCHERICHIA COLI
• EPEC – Non – toxigenic – diarrhea à children
• ETEC – Labile toxin (á CAMP)
Diarrhoea à Travellers
Stable toxin (á CGMP)
• EAEC – EAST – 1 toxin à Diarrhea à Travelers
• EIEC – Virulence Marker Antigen à dysentery
• EHEC – Verocytotoxin à dysentery & HUS (Hemolytic Uremic Syndrome)
(0157 : H7)

â Protein synthesis
• UPEC – fimbriae à UTI

B. Klebsiella 03:56

1) K. Pneumonia – pneumonia, Meningitis, UTI, wound infection

2) K. Granulomatis – Granuloma Inguinale a.k.a Donovanosis

Painless Red beefy genital ulcer

(bleeds on touch with Pseudobubos à Sub cutaneous nodules looking
like enlarged lymph nodes

C. Salmonella 06:40

Clinical picture – Typhoid fever à S. Typhi /

S. parayphi-A,B

Carriers – Stool carrier / Urine carriers

Diagnosis – Culture
à Wilson blair (selective for salmonella only)
à XLD, DCA, SSA (selective for both salmonella
& shigella)

• Test à Widal Test à Agglutination Test

• Carriers à
• vi Ab test à screening (persists for long in carrier state)

1
 • Stool/urine culture à Confirmatory
• Sewage swab culture à Community

. Shigella 0 :30
• Group A – Sh. Dysentriae (sever from of infection)
B – Sh. Flexneri (most common cause in India)
C – Sh. Boydii
D – Sh. Sonnei (most common cause in world)

Late lactose fermenter

• Clinical picture à Virulence marker Ag (VMA) & Enterotoxin à Dysentry

à Shiga toxin à (inhibits protein synthesis) à HUS
• Diagnosis à Culture à XLD, SSA, DCA

. Proteus 0:54
• Clinical picture à UTI, Wound infection, Pneumonia etc
Phosphate stones (urease) in urine (alkaline)

? Struvite Stones : Phenyl Pyruvic Acid, Phenyl Alanine Deaminase

• Diagnosis à PPA test – PAD Enzyme (only in proteus)Q

à Diene’s phenomenon

Proteus – Swarming growth

If swarming pattern of two strains merges, then both are of same strain

. Yersinia – Zoonotic Disease 4:50

1) Yersinia à Rat flea à Plaque (Bubonic, Pneumonic, Septicemic)

Pestes (Xenopsylla ?
(Non-motile) Cheopsis)

2) Yersinia Enterocolitica à Feco-oral à G.E

Differential motility ? Acquired from pig
3) Yersinia pseudo TB à Feco-oral à Izumi fever

2
• Diagnosis – Micro à Bipolar staining
(methylene blue) à

Safety pin appearance

à Culture à Ghee broth àStalactile growth

. Vibrio :30
• Non-Halophilic (NaCl – 0.5-1%)
• V. cholerae à V. cholera O1 (non-capsulated) à Pandemic & Epidemics
V. cholerae O139 (capsulated) à Epidemics
Oxidase +ve

V. Cholerae

• V/F à Cholera toxin (á cAMP)

• C/P à Cholera (Life threatening diarrhea) à Rice watery Appearance
• Diagnosis à Micro à Comma shaped with fish in stream appearance darting
motility in hang & drop

à Culture à TCBS à Yellow color à Sucrose fermenter

àTests – String test +ve
Cholera Red reaction +ve
Halophilic Vibrios

• Minimum 6% NaCl for growth

• V. Parahemolyticus
• V/F à Capsule
• Clinical picture à G.E, wound infection
• Diagnosis
• Micro – Bipolar staining à safety pin appearance
• Culture – TCBS à Green colony (Sucrose Non-Fermentes)
Wagatsuma agar à Kanagawa phenomenon

à Hemolysis by Para hemolyticus

3
 . Pseudomonas 5:50
• V/F à Exotoxin – A à inhibits EF2 & Protein Synthesis
• Clinical picture à Swimmer’s ear à children
àMalignant otitis externa à Diabetes
à Most common infection in burns cystic fibrosis patient
• Diagnosis à Culture à Cetrimide agar
• Non fermenter ( Do not ferment any sugar )
• PsuedomonasQ
• BurkholderiaQ
• AcinetobacterQ
• Brucella
• Bordatella
. Burkholderia :00
• B. Pseudomallei – Mallediosis

Safety pin appearance (motile, oxidase +ve)

• B. Mallei à Glander’s disease ( Zoonotic disease from horse )

(non-motile, oxidase -ve)

• B. Cepaciae à Nosocomial pneumonia

(motile, oxidase +ve)

. Campylobacter 30:00
• Clinical picture à diarrhea
• Diagnosis à Micro à ’S’ shape
à Culture à Skirrow’s medium

Helicobacter Oxidase +ve ?

• Clinical Picture à Diarrhea, PUD, Gastric Ca.

• Diagnosis à Micro à Comma shaped
à Culture à Skirrow’s medium
à Test à Rapid Urease +ve

4
Gram negative coccobacillus
A. Hemophilus

H. influenzae B

• M.C Pathogen in children

• Clinical picture à Pneumonia/meningitis/epiglottis
• Diagnosis à Micro – GNCB (Gram Negative Cocco Bacillus )
à Culture – Filde’s digest agar
à BA à Satellitism

Hemophilus ducreyi

• Sexually transported
• Chanchroid (soft chancre)

Painful genital ulcer à bleeds on touch

with bubos (lymphadenopathy)

Safety pin appearance on bipolar staining

B. Bordetella
• Gram Negative Coccobacilli
• Clinical picture à Wooping cough / 100 days cough
• Diagnosis à Microscopy – Thumb print appearance
à Culture – Bordet Gengue medium

• Bisected pearl colonies

• Aluminum pain appearance

C. Brucella 36: 0
• Gram Negative Coccobacilli
• Clinical picture à Brucellosis (cattle) a.k.a Undulant Fever à Reticulo Endothelial
System à Hepato-Splenomegaly, Musculoskeletal System, Arthralgia & Myalgia
• Diagnosis àMicro àGNCB
à Culture àTrypticase Soy agar
à Test à Standard Agglutination Test

5
Spirochetes – Spiral Shaped
A. Treponema Pallidum
• Clinical picture à Syphilis ?

1° 2° Latent 3°
Hard Papules Asymptomatic Severe
chancre Gumma
â â â â
Painless Condylomalata Serology +ve Destructive
within Bubos â lesions
(Enlarged On Skin Folds involving
L.N) Skin, S/C
tissue.
Muscle &
Bone
â
CNS, CVS

# Genital Ulcer in H. Ducryei à Soft Chancre

• Congenital Syphilis

Early Late
- Symptoms ≤ 2years of - Symptoms > 2 years of
Age Age
- Skin, Joint, RES Hutchinson’s triad
- Skin Symptoms : - Interstitial Keratitis
Rashes, papules, - SNHL
Vescicles & Bullae - Hutchinson’s teeth
- Joint Symptoms :
Arthralgia
- RES Symptoms àHS
Megaly

• Diagnosis à Micro – Dark field microscopy – Thin and slender

• Motility à Flexion – Extension type
• Silver impregnation staining (Fontana)
Brown color Spirochetes in golden yellow background
• Culture – Rabbit testes ( Animal Inoculation)
• Tests –
Non – Treponemal Treponemal
VDRL FTA – ABS (Gold Standard)
RPR TPHA
6
B. Borrelia 46:0
• Burgdoferi à Lyme’s disease (Erythema Migrans) àIxodes Tick
• B. Recurrentis à Relapsing fever à Louse
• B. Vincenti àVincent’s angina (abscess in Gingival Space) à Commensal à Injury
to oral cavity
• Diagnosis Culture à Modified Kelly’s medium

C. Leptospira
• Zoonotic Disease
• Clinical picture à Leptospirosis à R.E.SàHepatosplenomegaly, Renal
(Rat’s urine)

• Diagnosis à Culture – EMJH

à Test à MAT (Micrscopic Agglutination Test)

D. Rickettsia
• Black Eschar
• Endothelial Injury

Typhus Group

a) Epidemic Typhus àR. Prowazeki à Louse

à OX19 +ve
b) Endemic Typhus à R. Typhi à Flea
à OX19 +ve
Spotted fever group

a) Rocky Mountain à R. Rickettsiae àTick

à OX19, OX2, +ve
b) Indian Tick Typhus à R. ConorciiàTick
à OX19, OX2 +ve
c) Rickettsial Pox à R. Akarii à Mite
à -ve for OX19 & OXK, OX2

Non-Cultivatable Bacteria

• Mycobacterium Leprae (GPB)

• Treponema Pallidum (GNB)
• Rickettsia (GNB)
• Chlamydia (GNB)
7
Rickettsial Related Diseases
• Oreintia tsutsugumushi
â
Scrub typhus à Trombuclid Mite Larva
â
OXK +ve

• Bartonella Quintana • Coxiella burnetti

â â
Trench fever à Louse Q fever (inhalation)
â â
-ve for OX19, OX2, OXK -ve for OX19, OX2 & OXK

Chlamydia 5 :54

• Chl. Trachomatis TRIC à ABC à Trachoma

D–K à Inclusion conjunctivitis
à NGU (Non Gonococcal Urethritis)

• Chl. Trachomatis LGV àL1, L2, L3 àLGVà

Begawa Corpuscles inclusion bodies à Painless Genital ulcer within Bubos

• Chl. Psittaei à Psittacosis à LCL Inclusion Bodies

• Chl. Pneumoniae à Atypical pneumonia
• Diagnosis – Culture à Tissue culture

Mycoplasma 0 :00:00
• Cell Wall deficient bacteria
• Clinical picture à Atypical pneumonia
à NGU à Non Gonococcal Urethritis

• Diagnosis à Microscopy à Diene’s method

à Culture à PPLO àfried egg colony

Legionella

• Clinical picture àAtypical pneumonia

à Pontiac fever
à Myocarditis
8
• Diagnosis à Culture à BCYE agar
à Test à Rapid Urine Test (Highly Sensitive)

9
 Bacteria GPB
Gram POSITIVE BACILLI
1. Bacillus
1) B. Anthracis
• Virulence Factor – Anthrax toxin = acts by ­ cAMP
• Clinical picture – Anthrax à cattle
Cutaneous Anthrax à Hide Porter’s disease
à Leather industry
Malignant pustule àBlack Eschar
Pulmonary Anthrax (Wool Sorter’s disease)
Hemorrhagic pneumonia
• Intestinal Anthrax (Abdominal pain, diarrhea)

• Diagnosis – Microscopy à Bamboo stick appearance

à Non Bulging SporesQ

• Culture –
• Solid media – Medusa head appearance
• Solid media + Penicillin – String of pearl appearance
• Gelatin tube media – Inverted fir tree appearance

• Tests àAscoli thermo precipitation test à Detects the capsule of

B.Anthracis (Only bacterial capsule made of polypeptide)

2) B. Cereus à Gastro Enteritis

Emetic toxin Diarrheal toxin
(Staphylococcal â
? Enterotoxin) Canned meat & veg
Fried rice â
â Post formed
Preformed â
â IP 8-16hrs
Incubation â
Period = 1-6hrs Heat labile
â
Heat stable

1
2. Clostridium à (Spore producing) – Anaerobic Bacteria
1) Cl. Perfringens
• Virulence Factor = toxin
• Clinical picture à Gas Gangrene / Food poisoning
(myo necrosis)
Blisters/Bullae on the skin
Diagnosis àMicro =?

GPB = Subterminal/ Bulging spores à CLUB Shaped

àCulture = Robertson’s cooked meat

medium (Anaerobic Bacteria)

à Tests – Naegler’s reaction

( - toxin) Reverse CAMP Test

Double Zone Hemolysis

Common anaerobic infection

Spore forming Non – spore forming

â â
Clostridium Bacteroids
(B.Fragilis – Brain Abscess)

2) Cl. tetani = àGPB within Terminal

bulging spores (drum
stick app)
Toxin – Tetanospasmin

Presynaptic inhibition of inhibitory Neurotransmitters

(Glycine/GABA)

Spastic paralysis

Tetanus

2
 3) Cl. Botulinum
à Botulinum toxin
(A,B,C,D,E,F,G)7 8th
C2/Enterotoxin
â â
Presynaptic inhibition of Food poisoning
excitatory N.T (Ach)
âNeurotransmitters
Flaccid paralysis
(also known as botulism)

4) Cl. difficle
à Commensal in intestine
â
Pre-disposing factor
Longterm use of 3rd Gen Cephalosporin, Ampicillin,
Clindamycin, Proton Pump Inhibiters, AML, ALL
â
Exotoxin (A), (B)
â
Diarrhea & Pseudomembranous Colitis

T/t à Vancomycin + Metronidazole

Corynebacterium Diphtheriae 16:28

V/F à Diptheria toxin à inhibiting protein synthesis (EF2)

Clinical picture à Diphtheria
- Faucial (Pseudo membrane) àMost Common
- Cutaneous
- Systemic
Diagnosis – Micro àGPC = Chinese letter pattern/V shape or L shape

• Culture (Selective Media)

3
 > 6hrs (commensals will grow)
1) LSS à White colonies à < 6hrs (Early Growth)
Less selective
Metachromatic granules

• High 2) PTBA à Black colonies à 48 hrs to grow

Selective 3) Tinsdale à Black colonies with brown halo

• Tests – ELEK’s Gel ppt. Test (On Horse Serum agar) for Diphtheria toxin

Mycobacteria 21:

1) M. Tuberculosis
V/F – LAM
T.B
Mycolic acid
Clinical picture – Pulmonary TB
Extra pulmonary TB
Diagnosis – Micro – Direct immunofluorescence
(Best)
- ZN staining Short Pink Rods

- Culture
Liquid – MGIT (Take 3-4 weeks for growth)
Solid à LJ media (Egg media) (Take 6-8 weeks for growth)
Middle brook 7H1)/11 {Best Media}

‘2’ Acid fast bacteria

à Mycobacteria
à Nocardia

Test –
1) CBNAAT à Identification + Antibiotic Susceptible Testing for Rifampicin
2) Line Probe Assay àIdentification + Antibiotic Susceptible Testing for all drugs
3) Mantoux à Type IV Hypersensitivity Test à Skin induration
>10mm <10mm

+ve Intermediate or Negative

4
 4) Rapid à MPT 64 Ag

2) M. leprae

Classification
Ridley Jopling WHO
TT Tuberculoid (Pauci bacillary)à
BT àS≤5&N≤1
BB à (Lepramin-ve)
BL Lepromatous à (multi
LL àmost severe

à(S > 5 & N > 1)

à Skin test –
(Lepramin +ve)

S = Skin lesions
N = Nerve lesions

Clinical picture – Hypopigmented patches with thickened nerves

Lepra Reactions à Occurs during treatment ( Multi Drug Therapy )

Type – I Type – II
• Type – IV HSR • Type – III HSR
• At BB stage • At LL stage
• Th1 CD4 Response • Th2 CD4 Response
• Inflammation of • Crops of nodular
existing Lesions or lesions (ENL)
(New Lesions) • Rx – Glucocorticoids
• Rx – Glucocorticoids Or
Thalidomide

5
Atypical Mycobacterium 6:2

Runyon classification

Slow Growers (>1 week) Rapid Growers (<1 week)

Photo Scoto Non

chromogens chromogens chromogens

â â â
M. Kansasii M. Scroful MAC Fortuitum
accium
M. Chelonae
M. Marinum M. Szulgai M. ulcerans
abcessus
(Swimming (Buruli
pool ulcers)
granuloma)

Mycolactone toxin
Clinical picture – Lymph nodes
Cutaneous
Pulmonary

Actinomyces

• Filamentous bacteria
• Anaerobic Bacteria
• Clinical picture – Actinomycosis à discharging granules
(Known as –Lumpy Jaw) on Angle of Jaw

â
Yellow (Sulphur granules)
â
(Contains the actinomyces)
• Diagnosis – Microscopy à Sunray appearance
Culture à Thioglycollate broth
(fluffy balls at the bottom of the tube)

6
Nocardia 2:

• Clinical picture à Nocardiasis

(Most common) Pulmonary > Brain

Nocardiosis Abscess

(Most common)
à Actinomycetoma (cause of – Madura foot)
â
White – Yellow discharging
(Granules from the foot)

• Diagnosis – Microscopy –Sunray appearance

Culture – Paraffin bait technique

Listeria :2
•
Clinical picture – Listeriosis à Food poisoning (Source – Refrigerated food)Q

Lysteriolysin

- Neonatal meningitis (2nd most common cause)

• Diagnosis – Micro à GPB °
Hanging Drop à Differential motility
°
Tumbling agar

(Only Gram +ve

Bacilli showing differential motility)
• Selective Culture à PALCAM agar (Selective motility)

7
 Bacteria GPC & GNC
GPC GPB GNC GNB
Staphylococcus Bacillus • Meningococci Enterobactarales
Streptococcus Clostridium Gonococi • Escherichia
Pneumococcus Corynebacterium • Klebsiella
Enterococcus Mycobacterium • Salmonella
Actenomyces • Shigella
Nocardia • Proteus
Listeria • Yersinia
• Citrobacter
Spirochetes
• Treponema
• Borrelia
• Leptospira
• Atypical
Pneumonia
• Chalmydia
• Mycoplasma
• Legionella
GNC B Imp Oxidase +ve Non-Cultivable
• Hemophillus • Vibrio • Rickettsia
• Bordetella • Pseudomonas • Chlamydia
• Brucella • Campylobacter
• Acinetobacter • Helicobacter

GPC 01:30
1) Staphylococcus

• Virulence Factor:

1. Cell surface factor 2. Enzymes

â â
Protein A • Coagulase: Anti Thrombin like Activity
(Anti Phagocytic) (Local skin infection)
• DNAse ® Cellulitis
• Hyaluronidase
• Staphylokinase

# M.C in Abscess formation

1
3. Toxins

• Hemolysin® - Hemolysis
• TSST – Type – I TSS
• Enterotoxin G.E (Milk & Milk products)
• Incubation Period : 1-6 hrs
• Preformed
• Heat stable
• Exfoliative SSSS { Staphylococcal Scalded Skin Syndrome }
• Clinical picture Abscess, Cellulitis, Impetigo, Botryomycosis
* (most common)
• Native valve I.E & Nosocominal pneumonias
• Osteomyelitis
• Diagnosis Microscopy : GPC Clusters
à Culture – Blood Agar : b - Hemolysis
- Nutrient Agar à Golden yellow color (Oil Paint appearance)
à Test – Catalase +ve/Coagulase (+)

MRSA – Staphylococcus Aureus Resistant to all b- lactams antibiotics

!"#$%&'()*+$
- SA MRSA
,-(./ 1-$-

PBP PBP2a

SA: Staphylococcus Aureus, PBP: Penicillin Binding protein

§ Diagnosis – Disk diffusion Cefoxitin disc : 2nd Generation Cephalosporin

Phenotypic method

PCR à mec – A gene à genotypic method

§ Treatment : Drug of choice – Vancomycin (Glycopeptide) àDOC

Linezolid
Streptogramin ?
VRSA Daptomycin {Best/DOC}
5th gen Cephalosporins

2
 CONS – catalase +ve/coagulase –ve

CONS: Coagulase Negative Staphylococcus

Staph. Epidermidis

§ Skin à Injury à Stitch abcess (Surgical Site Infection)

Prosthetic valve I.E (< 12 months of surgery)
(most common)

# > 12months of Surgery à Streptococcus Viridans Sanguis

Staph. Saprophyticus

§ Commensal in Female urethra Injury (Sexually active)

UTI

Streptococcus 1 :00

§ Virulence Factors

1) Cell surface proteins à ‘M’ (antiphagocytic)

2) Enzymes
DNAase (streptodormase) à A,B,C,D
Cellulitis (most common)
Hyaluronidase
Streptokinase

3) Toxins
§ Hemolysin (streptolysin)
Streptolysin S Streptolysin O
Oxygen Oxygen
stable Labile

? b - Hemolysis on BA ASO (Ab)

Less immunogenic BA Highly Immunogenic

3
 § TSST – Type 2 à Toxic Shock Syndrome (Super Ag)
(Pyrogenic exotoxin)
§ Clinical picture à Based On Carbohydrate antigen (By Lancefield) à groups
(Only A & B pathogenic)

1) Group A
§ Pharyngitis à Acute Rheumatic fever à ASO Ab >200 IU/mL
§ Skin Infections : PSGNàAnti DNAase(Ab)>300IU/mL
Cellulitis
Impetigo
Erysipelas
2) Group B
§ Neonatal meningitis (agents B –GPB :Strep (most common)
E – E. Coli
L – Listeria
§ Diagnosis – Micro GPC (chains)

Culture àb. Hemolysis on Blood agar

Tests à Catalase –ve
Bacitracin sensitive (GP-A)
Bacitracin Resistant, CAMP test +ve (GP-B)

3) Pneumococcus

§ Clinical picture Meningitis (most common)

Pneumonia (most common)

§ Diagnosis Microscopy – GPC Pairs Lens shaped Capsulated

à Culture : - hemolysis with Carrom Coin appearance

After 1-2 days

à Test àCatalase –ve

Optochin sensitive
Bile soluble +
Inulin fermenter +

4
 à Vaccine – Capsular vaccine
‘23’ serotypes ?

PPSV – 23 àPneumococcal polysaccharide vaccine in Adults

• C/I in < 2yrs

• Less immunogenic
à Pneumococcal conjugated vaccine àPCV – 13
more immunogenic ‘13’ serotypes
- Can be given to < 2year old child
Faecalis – most common
infection in humans ?

4) Enterococcus
Fecium – most drug resistant

§ Clinical picture UTI (most common), wound infection,

intra abdominal infection

Diagnosis à Micro – GPC in pairs – spectable shaped (Non-capsulated)

Culture – No hemolysis (No hemolysin)
Test –
§ Catalase –ve
§ Bile esculin test +ve
§ PYR test +ve

GNC 31:

1) Meningococcus – Neisseria Meningitidis

§ Clinical picture Meningitis within rashes

à Waterhouse Friedrichsen syndrome

?
Bilateral adrenal Hemorrhage & multiorgan failure

§ Diagnosis Micro – GNC – Pairs – Lens shaped Capsulated

à Culture àThayer martin medium (Selective media for Neiserria)

à Test àMaltose fermenter

5
 § Vaccine –
a) Capsulated àC.I < 2 years
b) Conjugated à Can be given in < 2 years

2) Gonococcus à UTI – (Imp virulence factor - fimbrae)Q

§ Clinical picture

à Males à Urethritis aka Gonorrhoea

Water can
Perianal Abscess perineum

à Female àVulvovaginitis
Fitz Hugh
+
Curtis syndrome
Perihepatic inflammation
à Neonates à Ophthalmia neonatorum

§ Diagnosis Micro – GNS Pairs Kidney shaped Non capsulated

à Culture – Thayer Martin’s media

à Tests – Maltose Non – fermenter

6
 General Microbiology
Scientists
• Antonie Von Leeuwenhoek (Father of microscopy)
Simple microscope

• Louis Pasteur (Father of Microbiology)

• Vaccination (Cholera , Anthrax, Rabies)
• Sterilization (Pasteurization, Hot air oven & autoclave)
• Fermentation of sugars
• Germ theory of diseases
• Liquid culture media

• Robert Koch (Father of Bacteriology / Modern Microbiology)

• Solid culture media

• Isolated Bacteria (CAT) Cholera, Anthrax, TB disease causing bacteria

• Hanging drop (demonstrate motility)
• Staining technique (Aniline /Basic dye)
• Koch postulate

• Paul Ehrlich (Father of chemotherapy)

• Salvarsan (to treat syphilis)

• Acid fasting staining à modified by Zeil & Neison Technique
• Standardize toxin & antitoxin

• Ignan Semmelweis (Father of Infection control)

Hand washing

• Joseph Lister (Father of Antiseptic)

Disinfectant (Phenol/carbolic acid)

1
Sterilization & Disinfection 05:40

Sterilization – Killing all forms life including spores

Sterilant – Agents used for sterilization

Disinfection – Killing all forms of life except spores

Disinfectants – agents used for surface disinfection

Antiseptics - agents used for skin disinfection
Methods of Sterilization & Disinfection

1) Physical methods – physical agents are used

Sterilants
2) Chemical method – chemical agents are used

Sterilants/Disinfectants/Antiseptic
Physcial method

Heat Filtration Radiation

#
DEO
Dry Moist Dept Membrane ionizing Non-
ionizing
Hot air Autoclave Asbestos Membrane X-Rays
oven (121°C-15 mins-15psi) g Rays
(160°C-2hrs) IR rays
UV rays
• Tantalization (100°C – 20 mins)
• Inspissation (80°C – 85°C à 30 mins)
• Pasteurization
Holder’s (63°C – 30 mins)
(Coxiella à Not killed)
Flash (72°C – 15 secs)
# Mechanism of Actions

• Dry Heat : Denaturation of Proteins, Electrolyte Imbalance, Oxidative Damage

• Moist Heat : Denaturation & Coagulation
• Filtration : Filtration
• Radiation : Nucleic Acid Damage

2
physical methods

A. Dry Heat

Hot air oven

• Biological control à Bacillus Subtilis/ Bacillus atropheus

• Materials sterilizedà

Glass à Flask, pipette

Sharps à Scalpel, Scissors, forceps
Oil à Liquid paraffin
Powder à Glove dust powder

B. Moist Heat

Autoclave

• Biological control à Geo – bacillus stearothermophilis

• Materials sterilized

• Culture media – all except serum /Egg containing CM (inspissations >

Tyndallisation)
• Surgical instruments (Except Sharps) - all sutures except cat gut sutures
(ionizing radiation)
• Disposable Plastic & Rubber - Gloves, Foley’s catheter,Syringes

C. Filtration - Membrane filters are most commonly used

• Uses - Serum, vaccines, antibodies (Bacterial membrane filters)
- To isolate bacteria from viruses (Mask filters)
- Surgical masks & N95 masks (HEPA & ULPA filter)
- Biosafety cabinets
# Deep filters : like Asbestos not used nowadays as they are Carcinogenic

3
 D. Radiation
• Biological control à Bacillus pumilis spores

Material sterilized

Ionizing radiation (better) Non – ionizing radiations

High penetrating power (Low penetrating power)

X-Rays/g-Rays IR/UV – Rays

• Disposable (Rubbers & Plastic)
• Catgut sutures Biosafety cabinets

Chemical method

HLD ILD LLD

(spores +) (spores ±) (spores -)
• ETO (alkylating • Halogenic compound • Alcohol (cell membrane damage)
agent) (oxidative damage)
• Plasma (oxidative • Phenolic compound
damage) (cell memebrane
• Aldehydes damage)
(alkylating agents )

• à ETO (Ethylene oxide) • à Plasma (H2O2 à ions)

Biological control Biological control

Bacillus subtilis/Atropheus Geo-bacillus

Materials Sterilised Materials Sterilised ?

• Surgical instruments
• Disposable (Plastic & Rubber)
• Heart lung machine
• Ventilator
• Dialysis machine
• Dental Equipments

Highly inflammable
Highly toxic
Highly carcinogenic

4
 Aeration – 8-10hrs
(to remove residual ETO)

• Halogenic compound

Disinfectants (Cl2) Antiseptic (I2)

Sodium Hypochlorite Tincture I2

(0.5-1%) Povidone –I2

• Phenolic compounds

Disinfectants Antiseptics
• Lysol (floor) • Chloroxylenol (Dettol)
• Cresol (sharps) • Chlorhexidine (Savlon)

• Aldehyde à Disinfectant

• Formaldehyde (40%) - OT sterilization

( Gas ) (Not used now days)
• Glutaraldehyde (2%) – Endoscopes
• Ortho – Phthal – Aldehyde – Endoscope (better)

• Alcohols

Ethyl alcohol (70%) à hand rub, metal surfaces

Isopropyl alcohol (70%) à hand rub, thermometer

Tests for Disinfectants (reference disinfectant à phenol)

1) Rideal walker à efficacy à in absence of organic matter
2) Chick martin à efficacy à in presence of organic matter efficacy
3) Kelsey Sykes à capacity à to retain its efficacy even after repeatedly use
capacity
4) Kelsey Maurer’s in use test àcontamination à of an disinfectant

5
Spaulding classification of medical devices

1) Critical – penetrates mucous membrane àHLD

Or
Enters sterile site
Eg: IV – catheters, surgical instruments
2) Semi – Critical – comes in contact within mucous membrane à HLD
Or injured skin

Endoscopes
3) Non – Critical à comes in contact with intact skin

Stethoscope, Sphygmomanometer
IDL/LLD

Morphology & Growth of Bacteria

Has the ability to cause Genital tract infections

Spore state Survival

6
A. Capsule à demonstration

1) Microscopic methods
a) Negative staining à India ink& Nigrosin: Capsule appears as clear halo
b) Most common Fayden’s reaction à Methylene blue

Stains only polypeptide capsule

Bacillus Anthracis

2) Immunological method
a) Latex Agglutination Test (LAT) à by using anti capsular antibodies coated on
latex particles

Rapid test to detect capsule

b) Quellung’s Reaction

Detect only polysaccharide capsule :

Pneumococci
Meningococcus
H. Influenza B
Klebsiella

B. Cell wall demonstration

i. Electron microscope
ii. Micro dissection
iii. Plasmolysis

C. Flagella Demonstration 5:00

1) Direct methods à directly by

a) Electron microscopy
b) Flagellar staining (Tannic acid staining/ Leifson’s stain)
c) Silver impregnation staining

7
2) Indirect methods à demonstrate motility
a) Wet mount
b) Hanging drop
c) Dark field microscope
d) Semi solid agar
e) Phase contrast microscope
f) Anti H antibody

D. Fimbria à DEMONSTRATION :54

i. Electron microscope
ii. Hemagglutination

Spore – DEMONSTRATION :

i. Acid fast staining

ii. Ashby’s method
iii. Fluton’s method
iv. Ryu’s method

Bacterial Growth Curve ? :0

Microscopy & Staining 4 : 0

A. Microscopes à Magnifying instrument

Unaided eye à 0.2mm
Light microscope à 0.2µm
Electron microscope à 0.2nm (0.1nm – 0.5nm)
8
Types

1) Simple microscope à only 1 magnifying lens

2) Compound microscope placed at some distance à > 1 magnifying
Used now a days
a) Bright field microscope à Staining ?
(Light microscope) à Wet mount with Hanging drop (without staining)
b) Dark field microscope à No staining ?
(Dark field condenser)

Better to demonstrate motility of organism ?

c) Phase contrast microscope à No staining

(Phase Plate)

d) Fluorescent microscope à Fluorescent dye

e) Electron microscope à Heavy metal staining

B. Staining 4 :00

Kills the organisms and the visibility of organism

1) Gram staining – To differentiate bacteria into G +ve/ G –ve

I. Primary stain – 1min – ?

{Crystal Violet/ Gentian Violet/ Methyl Violet}
Wash
II. Mordant – 1min à Helps in penetrating 1°
(Grams I2) stains inside the
organism & fixed inside
the organisms

III. Decolouriser (Acetone) à2-3 sec

Or
acetone – alcohol à 10-30 sec critical step
Wash
IV. Counter stain – 1min –
(Saffranin) or (Dilute Carbol Fuschin)
9
2) Acid fast staining à to differentiate bacteria into acid fast & non-acid fast

I. Primary stain à 5mins à ?

(Concentrated Carbol Fuschin)

Wash Mordant

II. Decolorizer à 1min Intermittent heating

(H2SO4)

till fumes appear

III. Counter stain à 1min à

(Methyl blue / Malachite Green)

3) Albert’s stain à to demonstrate metachromatic granules ?

I. Albert’s A solution à 5mins

(Malachite green / Toluidine blue)

Wash (with albert’s B solution)

II. Albert’s B solution à 2mins

Albert’s I2 (Mordant)

4) Impregnation staining à to demonstrate thin organisms/structure

a) Tannic acid impregnation staining

Ex – Lefson’s stain

To demonstrate flagella

b) Silver impregnation staining

Ex – 1) Warthin starry staining

To demonstrate H. pylori

10
 Fontana stain

To demonstrate spirochete

5) Negative staining à to demonstrate capsule

a) India Ink Clear halo with

b) Nigrosin dark background

Culture media and culture methods 5 :00

Culture media

1) Based on O2 Requirement
a) Aerobic media – with presence of O2
Ex – Blood agar, Mc Conkey agar etc
b) Anaerobic media – with absence of O2
Ex – RCM broth, Thioglycolate broth

2) Based on consistency
a) Solid media – With 2% agar àBlood agar, Mc Conkey agar
b) Semi solid media – With 0.5 – 1% agar

Mannitol motility media

c) Liquid media – With no agar à Thioglycolate broth, alkaline peptone H2O etc.

3) Based on constituents
a) Simple/Basal media à Nutrient broth,
Nutrient agar
b) Special media – (special constituents added)

11
1. Enriched media àBlood agar, Chocolate agar
2. Enrichment media à(used only for stool sample) : All are liquid
Ex – • Selenite F Broth For all stool
• Tetrathionate broth vibrio pathogens except
vibrio
• Alkaline peptone H2O à for vibrio

3. Differential/Indicator media
Ex – Mc Conkey agar (MA) à Lactose Fermenting vs Non Lactose Fermenting

• G +ve are inhibited à only G –ve grows

• Swarming is promoted

- Cysteine Lactose Electrolyte Deficient media (CLED)

LF vs NLF
• Swarming is inhibited
• G+ve also grows à along with G-ve
4) Transport media
Ex – Pike’s media à Streptococcus
Amies/Stuarts media à Gonococcus
Clairy Blair media (better) à all stool pathogen
Buffered Glycerol Saline à all stool pathogens except vibrio & Campylobacter
5) Selective media
Ex – Mannitol salt agar – Staphylococcus aureus
Thayer martin media – Nisseria
Wilson Blair – Salmonella
TCBS – Vibrio
LJ media – M. TB

12
Culture Methods 0 :05:00

1) Routine culture methods

a) Streak culture ?

With inoculation loop

b) Lawn culture/carpet culture

With cotton swab

c) Stroke culture ?
With inoculation wire

d) Stab culture
With inoculation wire

2) Anaerobic culture methods : Removes O2

a) Mc Intosh filde’s jar
b) Gas pack
c) Glove box
d) PRAS
e) Anaxomat (automated)

IDENTIFICATION 0 : 0:00

1) Biochemical Reactions
2) Molecular methods
3) Immunological methods

1) Biochemical Reactions Chemical Reactions

• Catalase (+ve) • Oxidase (-ve)

• Indole, Mannitol motility, TSI, Citrate, Urease

13
 A) Catalase (-ve) (7) ? B) Oxidase (+ve) (12)

• Streptococcus • Meningococcus
• Pneumococcus • Gonococcus
• Enterococcus • Pseudomonas
• Cardiobacterioum • Vibrio
• Eikinella • Campylobacter
• Kingella • Helicobacter
• Shigella dysentriae • Hemophillus
• Type 1 • Bordetella
• Brucella
• Cardiobacterium
• Eikinella
• Kingella

C) TSI (Triple Sugar Iron)

Findings Salmonella Salmonella Salmonella

Typhi paratyphi A paratyphi B
Sugar fermentation + + +
(colour change)
Gas production - + +
(break in medium) anerogenic
H2S production + - +
(black) (Scanty) (abundant)

2) Molecular methods à detects the gene à PCR àPolymerase Chain Reaction

Steps in PCR
1) NA Extraction – Enzyme method à
(lysozyme)
2) NA amplification – Denaturation (95°c)
ds à ss
A n n e a l i n g ( 5 5 °c )
Attachment of primers
E x t e n s i o n ( 7 2 °c )
Polymerase extend strands

14
3) NA detection à Gel Electrophoresis (conventional PCR)
OR
Fluorescent method (Real time PCR)

Genetics 0 : 0

Antimicrobial Resistance (AMR) à due to resistant genes

Resistant genes are transferred by :

1) Horizontal Gene Transfer

2) Mutation

i. Horizontal Gene Transfer

a) Conjugation – by physical contact

Most common in all bacteria except staphylococcus

b) Transduction – by bacteriophage

Most common à staphylococcus

c) Transformation – free transfer of gene from one bacteria to other
d) Transportation – by transposons a.k.a Jumping gene
ii. Mutation à Mycobacteria

Antimicrobial Susceptibility Testing 0 :

Methods

i. Phenotypic methods

a) Kirby Bauer’s à Disk diffusion ?

15
 b) Dilution methods à Broth dilution (Gold std) : Liquid Culture Media

(Antibiotic dilution)

à Agar dilution

c) Epsilometer test -

ii. Genotype methods : detects gene which has resistance to Antibiotics

PCR à Ex – Mec – A gene

Detected à MRSA Not-detected MSSA

16
 General Virology
General Virology 00:40

DNA DNA RNA (env) RNA (PCR)

(env) (non-env) (RFT & Others) (non-env)
(HHP) (PAP)
Hepadna Papova (HPV) Retro (HIV) Picorna (Entero virus,
(HBV) polio, coxsackie, HAV)
Herpes (8) Adeno (adeno) Flavi (HCV, KFD, Calci (HEV)
JE Dengue, YF,
Zika)
Pox (largest) Parvo Toga Reo (ds RNA) (Rota)
(smallest) (ss (chikungunya
DNA) Rubella)

Others 8

Arena - Lassa fever

Bunya - Hanta , CCF
Orthomyxo - Influenza
Paramyxo - Para influ, RSV, mumps, measles
Nipah
Rhabdo - Rabies
Filo - Ebola, marburg
Delta - HDV
Corona - Corona

1
Symmetry 0:4

Icosahedral à H, H, PAP, RFT, PCR

Complex à Pox
Helical à Others – A, B, O, P, R, F, C, D
Segmental NA à

R I B A
Rota Influenza Bunya Arena
(11) (8) (3) (2)

Circular NA à

A B D H P
Arena Bunya Delta HBV HPV

Inclusion Bodies 4:00

Precipitation of components inside the cell during viral replication

Intranuclear

Torres bodies à Yellow fever

§ Cow dry Type A
Lipschultz bodies à HSV
Owl eye appearance

(CMV): Perinuclear Inclusion Bodies

§ Cow dry Type B à Adeno, Polio

Intracytoplasmic

• Rabies – Negri
• Pox –
Small pox – Paschens
Vaccinia - Guaneri
MC - HP {Handerson Peterson}
Fowl pox - Bollinger

2
 NOTE – Both Intranuclear + Intracytoplasmic

Measles

(k/s – Warttin Finkeldy bodies)

Virus Cultivation : 0

1) Animal inoculation à mice

Cerebrum Foot pad

Rabies, Coxsackie M. Leprae

(Non-cultivable bacteria)

Rabbit à Testes à T. pallidum

2) Embryonated Egg Culture

* Vaccines prepared in the allantoic cavity àYellow fever

Influenza
Rabies
3) Cell culture / Tissue culture

1. Primary cell line à Ex : Chick Embryo fibroblast

Rhesus monkey kidney
2. Diploid cell line à Ex : Human diploid lung
Fibroblast
(W.I-38& MRC-5)

3
 3. Continuous cell line à Ex : HELA, HEP-2, Mc Coy,
BHK, Viro
(most commonly used)

Viral Essays :

Quantification of viral cell lines

1. Pock Essay à CAM à HSU, Pox

2. Plague Essay à Tissue culture à All viruses
# Non cultivable viruses : Hepatitis

Vaccines 0:00

• Live à BCG, OPV, Rota, MMR à National Immunisation

Schedule

à Small pox, Chicken pox, Typhoid, Cholera

(Herpes virus - 3)

à J.E, dengue, YF, Influenza

§ C/I à Immunocompromised patient & pregnancy

As live virus can get back their pathogenicity if host is immunocompromised
due to mutation
• Toxoid – Diphtheria, Tetanus
• Sub unit Vaccine (Recombinant Vaccine) – Hepatitis B, HPV, Coronavirus
• In Corona Virus all are subunits / recombinant except COVAXIN : Killed Vaccine
• All other vaccines nit mentioned above are killed vaccines.

4
 Immunology
Immunity 01:00

Defense system of our body against foreign antigen into our body

Innate Immunity

• Present since birth

• 1st line defense mechanism (acts within 3-4 hrs)
• Non-specific
• No memory
Mechanism of innate immunity

Org à PAMP (Pathogen Associated Molecular Pattern)

Host cell receptor à PRR (Pattern Recognition Receptor)

(+) Ex – TLR (Toll Like Receptor)

Macrophage – phagocytosis
Host cells
¯ NK Cells – Cell lysis (by secreting perforins and granzymes)
Cytokines à IL-1, 6, TNF - a

Inflammation

Acute phase reactants

Complement – Alternate Complement Pathway

Lectin Complement Pathway

Lysis of organism ¬ Membrane ¬ Attack Complex

Acquired Immunity 04:05

• Acquired after the entry of pathogen

• 2nd line defense mechanism à 5-7 days
• Specific
• Memory
1
Mechanism of Acquired Immunity

activated activated
IL-12 IL-4/5

Cytolysis Th1/ Th2/

(CMI) CD4 CD4

IL-2, TNF- TNF-a B cells

Plasma cells
Macrophage à
Phagocytosis
Antibodies
NK cells à Cell lysis

CMI
Ag – Ab complex
Cell lysis
AMI/HI Complement (classical pathway)

• CMI – Intra cellular organisms

• AMI – Extra cellular organisms
Signals for T-cell activation
1) MHC –I/II to cell T-cell
2) Co – stimulatory signals
B7 to CD28
APC T-cell
3) Cytokine signal

2
MHC/HLA Antigens 1 :00

MHC/HLA genes à short arms of Ch-6

MHC – I MHC – II MHC – III

Region Region Region

HLA-A,B,C HLA-DP/DR/DQ HLA genes

MHC – I MHC – II MHC – III

Protein Protein Protein
(Receptor) (Receptor)

All nucleated Only on APCs

cells & platelets

Ag presentation Ag presentation Complement

to CD8 to CD4 factors

CMI HI HI

# RBC donot have MHC-I/II

Antigens 1 :4

3
Heterophile Ag Similar Ag on two different cells/organisms

Ex – EBV Ag & Sheep RBC Ag àPaul Bunnel Test

Rickettsia Ag & Peroteus OX –K,2,19

Weil-Felix Test

Antibodies 1: 0

IgG – Monomers
- Highest serum concentration
- 4 Serotypes G1
G2 – doesn’t cross placenta
G3
G4 – doesn’t fix complement

Functions
• Fix the complement ( Acquired immunity )
• Crosses the placenta à Immunity to the baby
• Precipitation & Neutralization Ag – Ab Reaction

4
IGM
• Pentamer (J-chain)
• Highest molecular weight
à Fix the complement
à Agglutination Ag –
Ab reaction

IgA – Types
Blood IgA à monomer
Secretory IgA à dimer (J - chain)
Secreted by mucous cells of Gastrointestinal, Respiratory,
Genitourinary Tract
Functions – secretion à breast milk
local immunity to the baby

IgD àIgD & IgM

(monomer)
Surface Receptor on B – cells

IgE (monomer) à Elevated in hypersensitivity – I/Helminthic infections

Antigen Antibody Reaction :00

1) Precipitation
Soluble Ag + Specific Ab à precipitate: Floating precipitates (floccules)
Ex – Tube flocculation à Kahn Test
T. pallidum
Slide flocculation à VDRL, RPR
Tube ppt à Ascoli thermo ppt à B. Anthracis
Plate ppt à ELEK’s Gel ppt à C, Diptherae
Immuno diffusion à Oudin à Single diffusion
In single dimension
Ouchterlony à double
diffusion in double dimension

2) Agglutination Tests (insoluble Ag + specific Ab à

agglutinates)
Ex – WAT (widal) – Salmonella
SAT (standard) – Brucella
MAT (microscopic) – Leptospira
LAT (latex) - Capsule
HAT (heme) – Fimbria

5
Heterophile agglutination test
Paul Bunnel à EBV
Weil felix à Rickettsia
Cold agglutination
Mycoplasma
Streptococcal MG

3) Neutralization
Ex – Toxin Neutralization
Eg – Naegler’s reaction à Clostrium perfringens (alfa toxins)
- Virus Neutralization – PRNT (Plague Reduction Neutralization Test) à
Viral Quantification

Immunodeficiency disorders :41

1) Humoral IDD (B. cell defect)

a) X – linked agammaglobinemia (Bruton’s disease)
Tyrosine kinase enzyme deficiency
x
Pre B cells B. cells No plasma cells No Antibodies

b) Combined variable deficiency

Intrinsic B-cell defect
x
B-cell Plasma cell x

c) Selective IgA deficiency

Cytokine defect
x
Plasma cell IgA x
2) Cellular IDD (T. cell defect)
a) Di George Syndrome
(Chr22q11) Thymic Hypoplasia
X
T cell maturation

6
 b) PNP deficiency disease (Purine Nucleoside Phosphorylase)
(Chr-14)
PNP ( )

Purine Hypoxanthine x

Purine accumulates

Purine is toxic to T-cells

c) Chronic mucocutaneous candidasis

?
T. cell defect – CMI deficient

3) Combined IDD (Both B & T. cell defect)

a) SCID (Severe Combined Immunodeficiency
Disease)
X – linked à IL – 7 deficient
X
Lymphopoetic growth
(T & B cells)
AR à ADA deficiency, JAK 3 mutation, RAG
mutation
b) Wiskott Aldrich Syndrome (Y - linked)

WASP Gene mutation

X
WASP Protein
Precursor
Lymphoid Lymphoid cells (T & B cells)
Cells X

c) Ataxia Telangiectasia

Kinase enzyme deficiency

?
T & B cells defect

7
4) Phagocytic disorders
a) Chronic Granulomatous disorder (X – linked/AR)
NADP oxidase enzyme deficiency

x
Oxidative burst
x
Phagocytic killing

b) Chediak Higashi disease (AR)

LYST Gene mutation

x
Phagosome – lysosome fusion x

c) Job’s Syndrome (Hyper Ig-|E Syndrome)

AD à STAT – 3 mutation
AR à DOCK – 8 mutation
x?
Neutrophil chemotaxis

5) Complement deficiency disorders

a) C1, C2, C4 deficiency à SLE
b) C3 deficiency à GN
c) C5b, 6, 7, 8, 9 (MAC) à Disseminated infections
d) C1 esterase deficiency à HAE
e) DAF deficiency à PNH
f) Factor I & Factor H deficiency à Recurrent pyogenic infections

Hypersensitivity 4 :00

1) Type – I Hypersensitivity
• Immediate type (20 – 30 mins)
• Extracellular Ag
• Humoral Immune Response (IgE)
• Example – Allergic Rhinitis
Allergic dermatitis
Anaphylaxis : ¯BP – Hypotensive Shock
¯
Systemic Allergy

8
2) Type – II Hypersensitivity
• Immediate type (2 – 3 hr)
• Cell bound Ag (RBC bound Ag)
• Humoral Immune Response (IgG)
Ex – Haemolytic Anaemia
Transfusion Reactions – ABO & Rh incompatibility
3) Type – III Hypersensitivity
• Immediate type (2 – 3 hr)
• Extracellular Ag
• Humoral Immune Response (IgG)
• Ag – Ab (IgG) complex
• Ex – Glomerulonephritis
Rheumatoid arthritis
Good’s pasture’s syndrome
Arthus Reaction
Serum sickness
4) Type – IV Hypersensitivity
• Delayed type (2 – 3 days)
• Intracellular Ag
• Cell mediated immunity (TDTH cell)
• Ex – Tuberculin Test
Lepromin Test

Transplant Immunology 5 :00

1) Graft Rejection
Immune
Recipient Graft
Response
a) Acute – weeks – months
Major à CMI (CD 8 T. cells)
Minor à HI (antibody)
b) Chronic - months – years
Major à CMI (TDTH cell)
Minor à HI (antibody)

9
 c) Hyperacute – mins to hrs
HI (pre-existing antibodies)

Previously received blood transfusion or Organ Transplantation from same

patient
Presentation - HLA compatibility testing
ABO compatibility testing
Immunosuppressive drugs

2) GVHS ( Graft Vs Host Disease ) Only in Bone marrow transplant

Thymus transplant
Graft Recipient

Immunocompetent Immunocompromised
T cells
(Bone marrow & Thymus)

a) Acute Within 100 days

b) Chronic after 100 days

Treatment : Glucocorticoids

10
Symmetry 10:41

Icosahedral à H, H, PAP, RFT, PCR

Complex à Pox
Helical à Others – A, B, O, P, R, F, C, D
Segmental NA à

R I B A
Rota Influenza Bunya Arena
(11) (8) (3) (2)

Circular NA à

A B D H P
Arena Bunya Delta HBV HPV

Inclusion Bodies 14:00

Precipitation of components inside the cell during viral replication

Intranuclear

Torres bodies à Yellow fever

§ Cow dry Type A
Lipschultz bodies à HSV
Owl eye appearance

(CMV): Perinuclear Inclusion Bodies

§ Cow dry Type B à Adeno, Polio

Intracytoplasmic

• Rabies – Negri
• Pox –
Small pox – Paschens
Vaccinia - Guaneri
MC - HP {Handerson Peterson}
Fowl pox - Bollinger

11
 NOTE – Both Intranuclear + Intracytoplasmic

Measles

(k/s – Warttin Finkeldy bodies)

Virus Cultivation 1 : 0

1) Animal inoculation à mice

Cerebrum Foot pad

Rabies, Coxsackie M. Leprae

(Non-cultivable bacteria)

Rabbit à Testes à T. pallidum

2) Embryonated Egg Culture

* Vaccines prepared in the allantoic cavity àYellow fever

Influenza
Rabies
3) Cell culture / Tissue culture

1. Primary cell line à Ex : Chick Embryo fibroblast

Rhesus monkey kidney
2. Diploid cell line à Ex : Human diploid lung
Fibroblast
(W.I-38& MRC-5)

12
 3. Continuous cell line à Ex : HELA, HEP-2, Mc Coy,
BHK, Viro
(most commonly used)

Viral Essays :

Quantification of viral cell lines

1. Pock Essay à CAM à HSU, Pox

2. Plague Essay à Tissue culture à All viruses
# Non cultivable viruses : Hepatitis

Vaccines 0:00

• Live à BCG, OPV, Rota, MMR à National Immunisation

Schedule

à Small pox, Chicken pox, Typhoid, Cholera

(Herpes virus - 3)

à J.E, dengue, YF, Influenza

§ C/I à Immunocompromised patient & pregnancy

As live virus can get back their pathogenicity if host is immunocompromised
due to mutation
• Toxoid – Diphtheria, Tetanus
• Sub unit Vaccine (Recombinant Vaccine) – Hepatitis B, HPV, Coronavirus
• In Corona Virus all are subunits / recombinant except COVAXIN : Killed Vaccine
• All other vaccines nit mentioned above are killed vaccines.

13
 Mycology
Fungi
Yeast Yeast like

- Cryptococcus - Candida
- Malassezia
- Tricosporon
Moulds -

(Hyphae)

Aseptate moulds (obtuse or right angle branching)

Rhizopus
Absidia
Mucor

Septate with acute < branching

Aspergillus

Septate with irregular branching

Dermatophytes

Dimorphic

Culture ?

• Histoplasma
• Blastomyces
• Coccidiodes
• Para coccidiodes
• Sporothrix Schenkii
• Penicillium Marneffi

1
Fungal spores 03:50

Sexual spores Asexual spores

Ascospores Arthrospores
• Septate moulds • Trichosporon
• Dimorphic fungi
Basidiospores Blastospores
• Yeast (Budding yeast like)
• Yeast like • Yeast
(except – C. albicans) • Yeast like
Zygospore Chlamydospores
• Aseptate moulds • C. albicans
• C. dublinensis
Conidiospores
• Septate moulds
Sporangiospores
• Aseptate moulds

Fungal disease (Mycoses) 05:25

1) Superficial mycoses (skin, hair, nails)

§ Tinea versicolor
§ Piedra
§ Dermatophytoses
2) Deep mycoses/subcutaneous mycosis (S/C)
§ Mycetoma
§ Sporotrichosis
§ Chromoblastomycosis
§ Pheohypomycosis
3) Systemic/Endemic mycosis (Respiration)
§ Histoplasmosis
§ Blastomycosis Cause
§ Coccidiomycosis Pneumonia
§ Paracoccidiomycosis

2
4) Opportunistic mycoses
§ Candidasis
§ Cryptococcosis
§ Aspergillosis
§ Mucormycosis
§ Pneumocystosis

Superficial Mycoses 0 :2

1) Tinea versicolor/Pytiriasis versicolor

Clinical picture Hypo/Hyper pigmented patches – Upper trunk
Organism Malassezia (M.Globosa) > M. furfur)
(lipophilic fungus)
Diagnosis SDA Fried egg colonies (culture)
Gram stain Spaghetti & meat ball appearance
Wood’s lamp Golden yellow fluorescence
2) Piedra : Nodules formation on hair shaft
White Piedra Black Piedra
Clinical picture – White Black coloured nodules on
nodule on the hair shaft the hair shaft
Organism – Tricosporon Piedra hortae
beijilli
Diagnosis – SDA white Brown – Black colonies
colonies Ascopores
GS/LPCB Arthrospores

3
 3) Dermatophytosis/Tineasis
Clinical picture dry skin with skin rashes (ring worms)
T. capitis – Scalp involved

Kerion – Boggy lesion – Scalp T. Verrucosum

(M.C)

Favus – cup like crust (Scutula) Hair follicle

T. Schoenleinii{M.C}

Ectothrix – Arthrospore formation Outside

hair shaft

(Most common) M.audonii

Endothrix – Arthrospore formation Inside hair

shaft

T. Tonsurans

Others: T. Corporis, T. Barbarae, T.Pedis etc. ?

Organism – dermatophytes Tricophyton - S, H, N S- Skin

Mirosporum - S, H H- Hair
Epidermophyton - S, N N- Nail

Diagnosis – SDA – White cottony growth

LPCB – Septate hypahe & conidia

Dermatophyte ?

Trichophyton Microsporum Epidermophyton

Microconidia -

Macroconidia

Wood lamp Apple green fluorescence (M. canis) most common

Hair perforation test +ve – (most common)
4
 T. mentegrophytes (M.C) & Microsporium canis

Deep Mycoses 5:00

1) Mycetoma
Clinical picture S/C swelling Sinuses Discharging granules
Organism Eumycetoma Actinomycetoma

Fungi Filamentous bacteria

Granules Granules

White Black White-Yellow Pink

Pseudollescheria Madurella - Nocardia
boydii mycetomatis asteroids
Actinomodura
pelleteri

2) Sporotricosis/Rose Gardner’s disease

Sub himalayan region
Clinical picture S/C nodules along lymphatics
Orgainsm Sporothrix Schenki (dimorphic)
Diagnosis Sample (PAS) – à

Culture (LPCB) – à
25°c
3) Chromoblastomycosis
Clinical picture S/C nodules, ulcers, verrucous lesions
Organism Pheoid fungi (darkly pigmented fungi)
Producing medlar bodies (sclerotic bodies)

Copper penny appearance

4) Pheohypomycosis
Clinical picture – S/C nodules, ulcers, verrucous lesions
Organism – Pheoid fungi do not producing medlar bodies

5
Systemic Mycoses Endemic Mycoses 20:00

1) Histoplasmosis
Clinical picture Pulmonary, cutaneous, RES
Organism Histoplasma capsulatum
Diagnosis Sample (PAS) Intracellular yeast

Culture (LPCB) Tuberculate conidia

(25°c)

2) Blastomycosis
Clinical picture Pulmonary, Cutaneous
Organism Blastomyces
Diagnosis Sample (PAS)

Broad base budding yeast

OR
Figure of 8 appearance

Culture (LPCB)
(25°c)

Pear shaped conidia

Along the Septate Hyphae
3) Coccidiomycosis
Clinical picture Pulmonary, cutaneous
Organism Coccidiodes
Diagnosis Sample (PAS)

Spherules
(containing Endospores)

6
 Culture (LPCB)
(25°c)

Arthrospores with fragmented Hyphae

4) Paracoccidiomycosis
Clinical picture pulmonary
Organism Paracoccidiodes
Diagnosis Sample (PAS) –

Mickey mouse app or Pilot wheel app

Culture (LPCB)
25°c
Pear shaped conidia
along the Hyphae

Opportunistic Mycoses 2 :00

1) Candidiasis
Clinical picture cutaneous, mucocutaneous, invasive
Agent Candida
Diagnosis SDA – white colonies
G/S

GTT Germ Tube Test ?

- C. albicans – (42°c – growth +ve)
- C. dublinensis (-ve) –(42°c – growth +ve)
BDG – marker for invasive candidiasis

2) Cryptococcosis
Clinical picture pulmonary, meningitis, cutaneous, invasive
Agent Cryptococcus
Diagnosis SDA – mucoid cream color colonies

7
 G/S - - Budding yeast cell

Negative Staining -

Capsule (only capsulated, fungi)

Bird seed agar - Brown color

(Melanin pigment)

3) Aspergillosis : Only septate showing acute angle branching. ?

Clinical picture – Pulmonary/para nasal sinus/ Ear/Invasive
Organism- A.Niger A.Flavus A.Fumigatus

Ear infection Aflatoxin (HCC)

Diagnosis - SDA – Black growth Yellow Green
LPCB -

1/3rd vesicle
Complete vesicle 2/3rd vesicle

Marker Galactomannan >> Beta D Glucan Marker of invasive aspergillosis

4) Mucormycosis
Clinical picture Rhinocerebral, pulmonary, cut,
invasive
Organisms - Rhizopus Absidia Mucor
(most common)
Diagnosis - SDA – White White cottony White cottony
cottony growth
with black dots
(salt & pepper
appearance)
?

8
LPCB - ?

5) Pneumocytosis
Clinical picture – pulmonary (PCP – Pneumocytis Carinii Pneumonia): earlier it was
considered as parasite (protozoa), now it sis grouped under fungi.
Organism – Pneumocystis Carinii/jeroveci
Diagnosis – SDA non – cultivable
PCR
Black cysts
Sample (GMS)-
Ping – poll
Ball appearance

9
 Parasitology - Helminthes
Morphology forms – worms (adult) à Eggs à Larva

• Cestodes (Tape worms) à T. Solium (Pork)

Flat & Tape like T. Saginata (Beef)
E. Granulosus (Dog)
H. Nana (Dwarf)
D. Latum (Fish)

• Trematodes/Flukes à Schistosoma (Blood)

Flat & Leaf like P. Westermani (Lung)
F. Hepatica (Liver)
F. Buski (Intestinal)

• Nematodes/cylindrical worms

• Parthenogenic worm – S. Stercoralis

• Hook worm –Ancyclostoma/duodenale/N. Americanus
• Round worm – A. Lumbricoides
• Pin/threadworm – E. Vermicularis
• Whip worm – T. Trichura
• Filarial worm – W. Bancroffi
B. Malayi
O. Vulvulus
Loa – Loa
• Spiral worm - T. Spiralis
• Guinea worm - D. medinensis

Cestodes 04:04

à Tape worms

a) T. Solium
Infection form
diarrhea, abdominal pain
Larva – man à Intestinal Taeniasis
(definitive host)
Egg – pig or man (accidental intermediate host) àcysticercosis
àAutoinfection

1
NOTE : In all the Helminthes having more than 1 host

Man is the definitive host

(Except – E. Granulosus – dog (definitive host)

Cysticercosis – Formation of tissue cyst containing larva

of T. Solium

- Deposited in – Brain, Eye, S/C, M/S

Neurocysticercosis Occularcysticercosis
?
(Most common) Parenchymal, Subarachnoid, Meningeal
spinal

Asymptomatic (Most common)

Seizures (Most common symptom)

Diagnosis – MRI brain à multiple small cysts with scolex

inside
Future head of adult worm

Treatment – Steroids {TOC} (1st) + Albendazole (next choice)

(-) Kill the cysts

Cause inflammation

Stool examination

2
b) T. Saginata (Beef Tape worm)
Infection form à Larva à Man (definitive host)

Man will never act as Intestinal taeniasis

intermediate host in à Egg à Cattle (Intermediate host)
T. Saginata
Cysticercosis
Diagnosis à Stool examination

Acid fast à Used to differentiate between T. Solium & T. Saginata

Egg is acid fast à T.Saginata

c) E. Granulous (Dog tape worm)

Dog à Definitive Host
Infection form à Egg à Man à Echinococcosis/
(int. host) Hydatid cystic
disease
Hydatid cysts are formed

Most commonly
undergo calcification
except - Lungs

3
 Diagnosis –
1) Fluid examination from tissue cyst post surgery
2) Imaging: CT Scan/USG + ELISA

(contains scolex)

Hydatid cyst Future head

3) Casoni’s Test à Hypersensitivity reaction /Skin test (not used now a days)

d) H. Nana (Dwarf tapeworm)

à Shortest Helminth
à Infection form à Man à Hymenolepiasis (abdominal pain & diarrhoea)
&
Only host Autoinfection

Only T. Solium and H. Nana

are the 2 cestodes, that
cause autoinfection

Diagnosis à Stool examination

e) D. Latum (Fish tape worm)

• Only cestode having 2 intermediate hosts
• 1st Cyclops
• 2nd Fish
• Infective form à Larva à Man (definitive host) à Diphyllobothriasis
àMegaloblastic anemia

4
 Diagnosis à Stool exam

à Operculated Egg

Only cestode – Causing megaloblastic anemia,

- Having operculated cyst
- Having two intermediate hosts

Trematodes (Flukes) : 4

1) Schistosoma (blood fluke)

S. hematobium S. mansoni S. japonicum
Larva Cercaria Cercaria Cercaria
(Infective
Form) Skin penetration Skin penetration Skin penetration

Bladder plexus Inf. mesenteric vein Sup. mesenteric

vein
Habitat

C/F - 1) Urinary Stool sample Stool sample

schistosomiasis
(painless terminal 1) Katayama fever 1) Katayama fever
hematuria) 2) Colon
(Diarrhea) carcinoma
2) Bladder carcinoma
Abdominal Pain,
Vomiting

Schistosoma Vs Other Trematodes ?

Larva – cercaria Larva – metacercaria
Intermediate host – one (snail) Intermediate host – 2
Transmission – skin penetration Transmission – Ingestion
Egg - non operculated egg Egg – operculated egg

Contains spine Does not contain spine

5
Nematodes : 0

à Cylindrical worms

1) S. Stercoralis (parthenogenic worm: ability of female worm to lay egg without male
worm)
à Only Helminth causing opportunistic infection
à Infections occur in immunocompromised
Larva (infective form)

Skin penetration

Cutaneous larva migran’s à (doesn’t transform into adult)

Loeffler’s syndrome àovo – vivi parous

Hypersensitivity in Lungs

(egg hatches immediately in intestine)

?
Q
Larva passed in stool sample

(Only Helminth presents as Larva is in stool sample)

2) A. duodenale à Old world hook worm

N. Americanus à New world hook worm
Also known as – Hook worm (one end is hooked in
intestine and suck the blood)

Causing iron deficiency anemia

Larva à Skin penetration à Cutaneous larval migrans

(infective form)
Loeffler’s syndrome

Ovo parous

Egg in stool sample

6
 (Q)
Blood fluke / S. stercoralis / Hook worm

Only 3 helminths, where larva enters via skin penetration. Rest all other example larva
enters via ingestion
à Egg -

3) A. lumbricoids
Egg à Ingestion à Visceral larva migration

(most common) Loeffler’s syndrome

Loeffler’s pulmonary Ovoporous

Eosinophillic
Egg in stool sample

(Q)
Most common cause of cutaneous larva migrans- Ancyclostoma brazilensis/Cannium
àAnimal Pathogen
(Q)
Most common cause of visceral larva mirgans - Toxocora cannis àAnimal Nematode

7
4) E. vermicularis (Large Intestine)
Egg à Ingestion à Peri – anal Itching à Eggs in NIH Swab à Perianal area
? (at night)

(Q)

Appendicitis, Autoinfection

Only Helminth causing appendicitis

Egg - Plano convex

5) T. Trichura (Large Intestine)

Egg à Ingestion à Chronic dysentery à Iron deficiency anemia
?
Rectal prolapsed
Eggs in stool sample

6) Filarial worm (viviparous) à filariasis

1. Subcutaneous Tissue à S/C nodules
2. Lymphatics à Lymphedema
Larva à Culex bite à Reach Habitat àViviparous à Filariasis à Larva in blood

Directly give birth to larva in subcutaneous tissue & lymphatics

Microfilaria larva in blood

Cutaneous Filariasis
Lymphatic Filariasis

Occult filariasis

• Larva is not seen in blood

• Larva is destroyed in viscera
• Also known as – Mayer Kuwenaar syndrome
8
• When microfilaria larva is destroyed in lung causing Hypersensitivity in lung

Called Tropical pulmonary Eosinophilia also known as Weingarten Syndrome

Lymphatic Filariasis Cutaneous Filariasis

W. bancrofti PBS O. Vulvulus

Microfilaria
Larva
Sheathed no nuclei No sheath
in the tail tip

B. malayi Loa Loa

Sheathed Sheathed
2 big nuclei in the Multiple, small
tail tip nuclei in the tail tip

7) T. Spiralis (Viviparous)
Give birth to larva
Encysted larva
Larva à Under cooked pork à Reach habitat à Muscle cyst
(infective form)

Larva in muscle in Biopsy

Most common muscle à Ocular muscle

Cardiac muscle à Myocarditis

Diagnosis – Bachmann intradermal test (Skin Test) à +ve

9
8) D. medinensis (Viviparous)
S/C

Larva à Easting Cyclops à Reach habitat à Viviparous à Skin blisters

containing larva

Non – Bile Stained Eggs 0 : 0:40

N – Necator Americanus
E – Enterobius Vermicularis
H - Hymenolepsis Nana
A - Ancyclostoma duodenale

Eggs floats in saturated salt solution 0 : :40

T – Trichuris Trichura
E - Enterobius Vermicularis
H – Hymenolepsis Nana
A – Ancyclostoma duodenale (Necator americanus)
A – Ascaris Lumbricoides (Fertilized egg)

10
 Parasitology - Protozoa
• Study of parasite is called Parasitology
Two parasites that cause infections to humans
1) Protozoa
2) Helminths

Protozoa à unicellular à 2 forms = Trophozoites à Cyst (Infectious form)

1) Free living (soil & water) (Trophozoites à cyst)
2) Intestinal (Trophozoites à cyst)
3) Vaginal (Ex – Trichomonas vaginals à only trophozoite and it is the infective
form)
4) Blood and Tissue (different morphological forms)

1. Free living protozoa (Amoeba : Protozoa with pseudopodia as locomotor organ)

à live in soil & H2O
a) Naegleria Fowleri
Clinical picture à PAM (acute) PAM : Primary Amoebic
Meningoencephalitis
(H/O-swimming)
Diagnosis à CSF mount

?
Cyst not seen in CSF
Treatment – Amphotericin – B

b) Acanthamoeba
Clinical picture à Granulomatous Amoebic Encephalitis (chronic)
Contact lens keratitis
Diagnosis – CSF mount

?
1
 Other – irregular
Inner – Polyhedral
Cyst are not seen in CSF

• All the free living protozoa are Amoeba

• All of them freely lives in soil& water à h/o of swimming
• All of them are Neuropathogenic : Acute condition à Naegleria
Chronic à Acanthamoeba Balamuthi
• Meninges involved – Naegleria
• Others à Encephalitis

c) Balamuthia

Clinical picture – GAE (chronic)

Diagnosis – CSF mount

d) Sappinia
Clinical picture – Encephalitis
Diagnosis – CSF mount

2
Intestinal Protozoa 10:45

1) Ciliates (cilia) – Balantidium coli

2) Amoeba (pseudopodia) – Entamoeba Histolytica
3) Flagellates (flagella) – Giardia Lambia
4) Sporozoa (Sporozoites) - Cryptosporidium/Cyclospora/Isospora

a) Balantidium coli (large intestine)

Clinical picture – dysentery

Diagnosis – stool mount

b) Entamoeba Histolytica (Large intestine)

Clinical picture – Amoebiasis types

Cyst in stools
Non – invasive intestinal amoebiasis – asymptomatic
(Cyst carriers)
(No invasion into intestine)
Invasive intestinal amoebiasis– flask shaped ulcer
(dysentery)

Caecum – most common site

Extra intestinal amoebiasis – abscess in Liver, Lung and Brain
Diagnosis – Non – invasive intestinal à stool mount à
(Tropho/cyst)
Invasive intestinal à stool mount à ELISA
Extra intestinal à Imaging (USG/CT) + ELISA
3
 Stool exam -

(Trophozoites)

Only Enntoamoeba Hystolytica is Trophozoite containing

RBCs (only in dystentry): not always present

Treatment –

Non-invasive intestinal • Paramo mycin

Amoebiasis (cyst carrier) • Iodoquinol
• Diloxanide Furate
Invasive intestinal • Metronidazole
Amoebiasis
Extra int à • Surgery + Metronidazole

To drain the abscess

c) Giardia Lambia (Small Intestine)

Clinical picture à asymptomatic, diarrhea, malabsorption

Diagnosis à stool mount
Badminton / Tennis racket shaped Trophozoite with Axostyle

Treatment – Metronidazole (Drug of choice for all intestinal protozoa)

• Sometimes; trophozoites/ cyst in the stool are not seen (as they are
? intermittently released)

Then duodenal wash done to collect the sample

Presence of trophozoite & cyst in duodenal wash is also diagnostic

4
 d) Cryptosporidium & Cyclospora & Isospora (Small Intestine)
No locomotary organ
Intestinal Sporozoa
• Have sporozoites
• No locomotary organs

Clinical picture à diarrhea

(Immunocompromised/opportunistic diarrhea)
Diagnosis à
Stool mount Acid fast staining

Vaginal Protozoa 0:40

a) Trichomonas Vaginalis
Flagellate
No Cystic Stage

5
Clinical picture – Vaginitis à itching, thin yellowish green discharge: Fishy smell
?

O/E à strawberry vagina

Diagnosis – Urine/vaginal discharge mount

Jerky motility

b) Bacterial vaginitis

• Gardenella vaginalis
• Thin white discharge
• Fishy smell, itching
• Microscopy à “clue cells” GNB with vaginal Epithelial cell

• Gram Negative Bacilli

c) Candidal vaginitis

• Candida albicans
• Severe intense itching
• White curdy discharge
• No fishy smell, Whiff test : Not positive
• Gram +ve Budding yeast cell with pseudohyphae

Blood & Tissue Protozoa :40

1) Flagellates – (Flagella)
a) Leishmania
b) Trypanosoma

6
2) Sporozoa – (Sporozoites)
a) Plasmodium
b) Babesia
c) Toxoplasma

a. Leishmania
Morphological forms

1) Amastigote –

• Intracellular
• Present within the macrophage
• No flagella
2) Promastigote –

• Spindle shaped
• Flagella present
• Infective form to human
Vector – Sand fly (phlebotomous argentepes)

NOTE : All the blood & tissue protozoa are transmitted by vector except Toxoplasma
(Feco-oral route)
Clinical picture – Leishmaniasis (L): M.C in India
• Cutaneous Leishmaniasis à L.Tropica (also known as – Oriental
Sores/Delhi Boil/ Baghdad Button )
• Mucocutaneous Leishmaniasis à L. Brazilensis
(Espundia)
• Visceral Leishmaniasis (also known as Kala Azar) à L. donovani
Complications: Post Kala-azar Dermal Leishmaniasis (PKDL)
Skin manifestations over face, which are pink coloured nodules after
visceral leishmaniasis
Diagnosis
PBS à Amastigote (LD body) within Macrophage
Test à Napier’s aldehyde test (detect increased IgG)à Hypergamma globinemia
(­IgG)
+ve only after 3 months of infection ?
à Rapid Test (detects rK39 Ag)
7
 àMontenegro test (Hypersensitivity test) à Positive

Treatment à Amphotericin – B (Drug of choice)

b. Trypanosoma

1) Tryp. Cruzi
• Morphological form à Trypomastigote (infective form)
Recurrent flagella

• Vector – “Reduvig bug”

• Clinical picture – Chagas disease à Romana’s sign (unilateral conjunctival
edema/eyelid edema)
Enlargement of tongue/esophagus/colon/heart
Clinical diagnosis ?

• Diagnosis – PBD à Trypomastigote

• Treatment à Benznidazole

2) Try. Bruci

• Morphological form à ? Trypomastigote

• Vector à Tse – Tse fly

• Clinical picture àSleeping sickness à “winter bottom sign” : Lymphadenopathy in
the posterior triangle of neck :
Clinical diagnosis
• Diagnosis à PBS – Trypomastigote
• Treatment à Suramin

Blood and Tissue Sporozoa 50: 0

a) Plasmodium (5 species)
Species (5)

P.ovale/P.falciparum/P.vivax/P.knowlesi/P.malariae

8
 Morphological forms
Sporozite à Trophozoite à Schizont à Gametocyte
(infective form) Liver, RBC

Vector à Female anopheles’ mosquito

Clinical picture à Malaria (Fever, Chills, Rigor), Hepatosplenomegaly

Diagnosis – PBS à Trophozoite, Schizont, Gametocyte

Trophozoites

P.vivax –

P.falciparum –

Single Multiple Accole Ring forms

Ring Ring forms

Ring attached to the

surface of RBC

Schizont

Not seen in Peripheral blood

# Schizont of P.Falciparum are adherent to endothelium of blood vessel

Gametocyte

Banana/crescent/sickle shaped : P. Falciparum

• Rapid test –
Plasmodium – PLDH Ag +ve (for all the species of plasmodium)

9
 Falciparum – HRP -2 Ag +ve

Histidine Rich Protein – 2

• Treatment – P.vivax à Quinine à Primaquine x 14 days (to prevent relapse)
P.falciparum à Artemisinin compounds

b) Babesia
• Morphological forms à Sporozoites à Trophozoites à Gametocytes
(infective form)
• Vector à Ixodes Tick/Hard tick
• Clinical picture à Babesiosis à Fever, Hepatosplenomegaly, Abdominal Pain,
Diarrhoea
• Diagnosis à PBS à Gametocyte

Maltese cross (diagnostic)

• Treatment à Atovaquone

c) Toxoplasma
• Morphological forms à Sporulated oocyst (infective form) à Tachyzoites

• Transmission à feco – oral route

• Clinical picture à Toxoplasmosis à Cervical lymphadenopathy

à Congenital toxoplasmosis

Triad – Chorioretinitis
- Hydrocephalus
- Intracranial calcification
• Diagnosis à PBS – Tachyzoites

à Gold standard test à Sabin Fieldman dye test

• Treatment – Pyrimethamine + Sulfadiazine
- Spiramycin (pregnant women)
• Drug of choice – Cotrimoxazole (In immunocompromised patient)
10
 Virology - RNA Viruses
HIV 01:00
Clinical picture  Stages (4)

Stage 1 – asymptomatic & Generalised lymphadenopathy

Stage 2 – Superficial fungal infection (skin)

Reactivation of viral infection
(ie – varicella zoster)

Stage 3 – Oral candidiasis

Oral Hairy Leucoplakia
Pulmonary TB
Fever > 1 month
Diarrhea > 1 month
Weight loss > 10%

Stage 4 – Oeseophageal candidiasis

Extrapulmonary TB
Wasting syndrome
Kaposi Sarcoma
Opportunistic infection

Diagnosis 02:50

Indirect – Antibody Tests (3 different rapid tests)

Strategy 2B Strategy 3

Symptomatic Asymptomatic

 

2 +ve All 3 +ve

Direct tests  RNA PCR  HIV gene (High sensitivity)

P24 Ag Essay

Window period  3 weeks – 3 months

 Abs are not detected
 Direct test  RNA PCR
1
Infant born to HIV +ve mother  direct tests  DNA
PCR

6 weeks after delivery/

6 months after delivery

Final diagnosis – after 18 months of delivery

do – indirect tests

Treatment – HIV – 0 :50

1st line L E T

(Lamivudine) (Efavirenz) (Tenofovir)

2nd line L L T

(Lamivudine) (Lopinavir) (Tenofovir)

HIV – 2 L L T

(Lamivudine) (Lopinavir) (Tenofovir)

PEP L L T

(Lamivudine) (Lopinavir) (Tenofovir)

L  NRT Inhibitors
E  Non NRT Inhibitors

Flavi 1 :10

i. KFD. V
Rat (Reserviour)  monkey  Ixodes ticks
(amplifier)
Humans

C/F – viral hemorrhagic fever

Vaccine – killed

2
ii. J.E
Herons/ pig  culex  humans
(ducks) (amplifier)

Encephalitis (M/C-up)
(age – 5 to 15yrs)

Vaccine – live (SA – 14-14-2)

Killed (Nakayama & Beijing strain)
iii. Dengue (5 - serotypes)
Aedes agypti  human  classical dengue fever (vector)
(C/F – Hemorrhagic fever & Retro orbital
Headache & Body pain)
Dengue Hemorrhagic fever Reinfection
with other
Dengue shock syndrome serotype

Vaccine – Live (CYD - TDN)

Due to antibody
dependent
Enhancement
iv. Yellow fever virus (Not reported in india)
Aedes  Humans  Hemorrhagic fever with
Hepatitis
Vaccines – live – 17 D strain
killed – Dakar strain

v. Zika virus
Aedes/ sexual route  Humans  GBS
Congenital  Microcephaly

(Q) Most common associated clinical manifestation with

congenital zika virus  Microcephaly

3
Toga Viridae Family 2 :50

i. Chinkungunya V (E1-226A mutation E1-226V)

Re – emergence of
chikungunya
Aedes  Humans  Arthritis

ii. Rubella virus

Clinical picture  adults – forschemier spots on
the soft palate

Small Bleeding sots (also known as -

Petiechial spots)

 Congenital

Salt & pepper retinopathy, cataract

CRS Sensory neural hearing loss
Patent ducts anteriosus

BMS  Petchial spots (all over the body)

Blueberry Muffin’s Syndrome

NOTE – If transmission to foetus occurred within the 1st

Trimester of pregnancy

The baby will develop congenital Rubella syndrome

- If transmission occurs after 1st trimester

(ie – organogenesis complete)

The baby will develop Blue berry muffin’s syndrome

4
RNA Non – Enceloped Viruses 2: 2

i) Picorna
Rhino V  Inhalational & Entero V  Feco – oral

Entero virus

a) Polio virus (serotypes)

Type 1  most common associated with paralysis
Type 2  vaccine derived polio virus – most common
serotype undergoing mutation in the vaccine
Type 3  vaccine associated paralytic polio
C/F – Polio disease  meningitis  1%
 flaccid paralysis <1%
Vaccine – live  Sabin (OPV)

Bivalent Killed  Salk (IPV)

(Type – 1,3)
Inactivated polio virus  I/M

b) Coxsackie V
Coxsackie A  Herpangina & HFMD
Coxsackie A – 24 serotype  Acute Hemmorrhagic
conjunctivitis
Coxsackie B  Myocarditis

c) Entero virus
Entero V 71  Herpangina & HFMD
Entero V 70  Acute Hemmorrhagic conjunctivitis

d) Hep – A. virus  acute hepatitis

Early – (Entero V 72)

e) Calci
HEV – acute hepatitis
Noro & Sappo – Gastro Enteritis

5
f) Reo
Rota (GIP[8])  G.E in children

Intususseption
Vaccine  live (oral)

Other Viruses 5:10

1) Arena – Lassa fever virus (Not reported in India)

- Rat (Reserviour)  Humans

Hemmorhagic fever & pneumonia

2) Bunya – Hanta V (Not reported in India)

- Rat (Reserviour)  Humans

Hemorrhagic fever & pneumonia

3) CCF V – Ixodes Tick (vector)  Humans

Hemorrhagic fever & pneumonia

4) Orthomyxo virus animals

Influenza virus  serotypes (A,B,C,D)

H1N1  swine flu/pandemic

Inf – A Sub serotypes H3N2  seasonal flu
H5N1  avian flu

B/O – Hemagglutinin receptor

&
Neuramidase Ag – attachment

Clinical picture – Pneumonia

Pandemic & Epidemic (due to antigenic variations)

Inf - B
Clinical picture – pneumonia associated with diarrhea
Epidemics
6
Antigenic Variations 5 :00

Antigenic shift – Genetic reassortment  Pandemics

Antigenic shift – Point mutation  Epidemics
Treatment – Neuraminidase inhibitors
Oseltamivir & Zanamivir
Vaccine – live (inhalation)
killed (I/M)
Diagnosis – ELISA

Ag detection

(Hemagglutinin Ag)

5) Paramyxo virus
a. Parainfluenza – Laryngotracheo bronchitis
b. RSV – Broncholitis in children
c. Humps – Salivary gland swelling
Meningitis
Gonaditis
d. Measles – most common – otitis media
Late – S.S.P.E  7-10yrs after 1° infection of
measles virus
4-10/1 lakh

Progressive mental detoriation with myclonus

Immunocompromised

(Hetch’s phenomenon)

e. NIPAH
Fruit Bat (Reserviour)  Humans  Encephalitis

Outbreak in India
2007
West Bengal
2011
2018 – Kerala

7
6) Rhabdo
a) Rabies virus
Animal bite Dog - India
Bat – India
Spread via Nerves

Brain stem encephalitis

Prophylaxis (100% mortality – No treatment)

Local – Wash wound  Running tap water = 15mins
- Antiseptic
- IG  Surrounding wound

Systemic – IG  im
- Vaccines

Regimens

PEP -0 3 7 14-28th day

Im -1 1 1 1
(Essen)
Id 2 2 2 -
(Redcross)
(0.1mi)
i.m 2 - 1 1
(Zagreb)
Pre P  0 7
I.m  1 1
i.d  1 1
PEP in 0 3
Pre P
i.m 1 1
i.d 2 2
i.d 4

8
7) Filoviridae family (filamentous)
Ebola & Marburg V (Not Reported in India)

Fruit bat (Reservoir)

Humans

Hemmorrhagic Fever

Delta
HDV Acute & Chronic Hepatitis
Co-inf & Super – inf with Hep-B virus
8) Corona
Corona virus (7)
SARS – CoV  2003  China  Bat
MERS – CoV  2012  Saudi Arabia  Camel
SARS – CoV – 2  2019  China  Bat
Clinical Picture – Fever, sore throat, cough, diarrhea,
myalgia
Pneumonia
IL-6
MOF (cytokine storm)

Diagnosis – Nasopharyngeal swab

Or
Oropharyngeal swab
PCR (more sensitive)
Direct Test
Rapid Ag detection
(less sensitive)

Treatment – Corticosteroids
& Tocilzumab
IL – 6 Inhibitors
Sarilumab

9
Vaccine – Killed  Covaxin
Subunit or
Recombinant

(‘S’ Ag)

10