Systemic Bacteriology

Systemic Bacteriology
• Bacteria can be broadly divided into:
• Aerobic bacteria – bacteria which require O2

for growth.
• Anaerobic bacteria – bacteria not requiring O2
for growth.
AEROBIC BACTERIA
• Gram positive
Cocci - e.g. Staph, Strep., Pnemococci, Enterococci
Bacilli/Rods – Corynebacterium, Bacillus,
Mycobacteria
• Gram Negative
Cocci - e.g. Neisseria
Bacilli/Rods - e.g. Enterobacteriaceae,
Pseudomonads, Pasteurella, Francisella
Gram
Positive
Cocci Bacilli/Rods
+ve CATALASE -ve
Strep Bacillus
Staph Pneumococci Corynebacterium
Enterococci Mycobacteria
Gram negative
Bacilli/ Rods Cocci
-ve +ve
OXIDASE
Pseudomonads
Enterobacteriaceae
Pasteurella
Francisella Neisseria
Gram Positive Cocci
GPC
• Can be divided into two types:
1. Catalase positive – Staphylococcus species

2. Catalase negative – Streptococcus species,
Strep. like (Pneumococci, Enterococci)
Staphyloccocus
Taxonomy
• Staphylococci, gram-positive cocci in the family
Micrococcaceae, form grapelike clusters on
Gram's stain.
Morphology - Gram positive cocci , size is 0.5-
1.5μm,. Occur singly, in pairs, tetrads, short
chains, and irregular grape like clusters.
Catalase positive
• Staphylococcus are differentiated from
micrococci by Hugh Leifson’s oxidative
fermentative test. Staphlococcus is fermentative
Media
• 5% sheep blood agar (SBA) – shows beta haemolysis
• MacConkey agar – pink colonies – means Lactose
fermenting
• Selective medium
– Selective salt media (10% NaCl)
– Mannitol salt agar-- selective & indicator with 7.5%
NaCl
– Ludlam’s tellurite media
• Selective media are useful for isolation of
Staphylococcus from food, faeces & where mixed flora
is expected
• Media for pigment – Milk agar, Glycerol monoacetate
agar
S. aureus on NUTRIENT AGAR
pigmentation is golden
• Mac- LF
SELECTIVE COLONY
MEDIUM MORPHOLOGY
MANNITOL S. Aureus : 1-3mm

SALT AGAR in dia, yellow
(Differential colonies surrounded
medium) by yellow medium.
S. epidermidis: form
small orange
colonies surrounded
by red or pink
medium
LUDLAM’S Staph: black
MEDIUM: colonies
Licl2+ telurite
Golden yellow pigmentation (due to carotene)
Media for pigment
– Milk agar,
Glycerol monoacetate agar
• Tween agar, incubation at 22˚c

IDENTIFICATION
• Characteristic colony
• morphology.
• CATALASE TEST:
– To differentiate from streptococci,&enterococci
– Detects the presence of cytochrome oxidase enzyme.

IDENTIFICATION OF S. aureus
• SLIDE COAGULASE TEST:
o Most strains of S. aureus have a bound coagulase enzyme

or ‘clumping factor’ on the surface of the cell.
o This factor reacts with fibrinogen in plasma causing rapid

cell agglutination.
• TUBE COAGULASE TEST:
o Detects free coagulase which is

secreted extracellularly.
o Reacts with CRF in plasma to

form a complex, which in turn
reacts with fibrinogen to form
fibrin.
What is a superantigen?
It causes non-specific stimulation of T cells leading to
cytokine release and inflammation which leads to
fever, hypotension and shock.
Superantigens –
• Act on Vβ of TCR
• Dont require antigen presentation by macrophage, directly stimulate non
specific Tcells
• Leads to massive release of cytokines & polyclonal B cell activation
Example :
• Staph- TSST, Exfoliative toxin, Enterotoxins.
• Strept- Str pyrogenic exotoxin A, B, C
• Mycoplasma arthritidis
• Yersinia enterocolitica
• Virus – EBV,CMV, HIV, Rabies nucleocapsid
• Fungus- Malassezia furfur
19
VIRULENCE FACTORS
Antigenic structure & virulence factors
Cell wall Activity
associated
Structures
Capsular Inhibits opsonisation
polysaccharide
Peptidoglycan Confers cell rigidity & induces inflammatory
response
Techoic acid Helps in adhesion to mucosal surfaces
Protein A Antiphagocytic, anti-complementary,
chemotactic
Binds to Fc of IgG leaving Fab free to bind to
Ag –Basis of Co agglutination
Clumping factor/ Responsible for slide coagulase reaction
Bound coagulase
• PROTEIN A:
o Has a unique ability to bind to Fc region of all human IgG

subclasses except IgG3 .
o Biological activities- interferes with opsonisation and ingestion

of orgs by PMN’s, activates complement, elicits immediate &
delayed hypersensitivity rxns.
o Is the basis for coagglutination test , used in labs for org

identification(eg:gonococci, streptococci).
o Absent in coagulase negative Staphylococci (CNS)

VIRULENCE FACTORS:HAEMOLYSINS
PROTEIN BIOLOGICAL GENE SPS
ACTIVITY LOCATION DISTRIBUTION
Alpha Haemolysin inactivated at 70C Chromosome S. aureus

reactivated paradoxically
at 100C
Beta Haemolysin Neutral sphingomyelinase. Chromosome S. aureus

Acts as a phosphatase, Is S.intermedius
haemolytic
(goat, cow& sheep> human.
HOT & COLD
PHENOMENON
ACTIVITY LOCATI DISTRIBUTION
ON
Delta Lyses rabbit Chrom S. aureus
Haemolysin sheep and osome S. epidermidis,
human red blood S.
cells haemolyticus
Gamma Lyses rabbit, Chrom S. aureus
Haemolysin sheep and osome
human red blood
cells
Leucocidins / Damange
Panton polymorphonucl
valentine ear leukocytes &
toxin macrophages
Furunculosis
VIRULENCE FACTORS: TOXINS
ACTIVITY LOCATION DISTRIBUT
ION
EXFOLIATIVE Have proteolytic activity. Dissolve the ET A is S. aureus
TOXIN A& B mucopolysaccharide matrix of chromosomall
epidermis, resulting in intraepithelial y encoded
splitting.
Intraepidermal blistering leading to ET B is
SSSS, Reiter in neonate, TEN in old,. plasmid
Has superantigen activity encoded.
ENTEROTOXIN Heat stable responsible for Variable S. aureus

A through E, H, I Staph food poisoning.
Type A mc. Ingestion of preformed exotoxin in
milk food - VAGAL STIMULATION, -
N/V with/without Diar. in 2-8hrs. Is a
Superantigen.
•
PROTEIN BIOLOGICAL SPS
ACTIVITY DISTRIBUTIO
N
TOXIC SHOCK Formerly k/a enterotoxin F. heat stable. S. aureus
SYNDROME Is a superantigen, leading to a systemic Some CONS
TOXIN-1( TSST -1) release of a variety of cytokines which is also
the cause of multisystem involvement in
TSS.
Is pyrogenic bcoz of IL-1 production,
causes erythroderma and endotoxic
shock.
Rashdesquamation & Multi organ failure

T/T- Clindamycin (reduces toxin synthesis)
Epidermolytic toxin - Mainly belong to phage group II
(Exfoliative toxin) - Two types; A: 30,000 daltons, heat stable, chromosomal gene
- B:29,500 daltons, heat labile, Plasmid controlled
Enterotoxins - Produced by 40% of clinical isolates

- Cause food poisoning
- Heat stable (not destroyed after heating food)
- Nine antigenic type (A, B, C, D, E, G, H,I, J)
Toxic shock syndrome toxin - Most strains belong to phage group I
- 22,000 daltons
- Earlier called as enterotoxinsF
Extracellular enzymes Activity
Coagulase Enzyme - Enzyme is secreted into the surrounding
- Requires coagulase reacting factor (CRF) present I plasma for
action
- Responsible for tube coagulase test
Staphylokinase - Breaks fibrin clots & allows spread of infection
(fibrinolysin)
Hyluronidase - hydrolyses Hyaluronic acid; facilitating spread of infection
Deoxyribonuclease - Degrades DNA, heat stable nuclease reaction
Lipase, Phospholipase, - Degrades lipid, Phospholipids, Degrades proteins
Proteases
• Mechanisms – resistance
1. Penicillinase production- (4 types –type4
mc)(plasmid mediated)
2. Altered PBP – methicillin resistance in staph
d/t this .( mec A gene)
3. Development of penicillin tolerance
MRSA  Methicillin resistant Staphyloccoccus
aureus
• (1961) - multi drug resistant
– important hospital pathogen
– Resistance is chromosomally mediated (mec A gene )

– Due to altered PBP 2a (MC) > hyper production of β lactamase
– Detection - 5g methicillin / 1g oxacillin disc

– cefoxitin
• - 4% NaCl, 30- 350C incubation

• Community acquired-( mec A IV)& PV Toxin,
• Hosp acquired -(mecA I,II,III)
• Detection - using methicillin /oxacillin disc /cefoxitin disk
- on Screening agar media containing 4% NaCl
- incubation at lower temperature of 300 ,
- full 24hr incubation

• Treatment- Vancomycin is DOC
– Others- Teichoplanin, linezolid, streptogramin,
– cipro/levofloxacin, cotrimoxazole, clindamycin, mupirocine,

minocycline
– Newer – daptomycin, ceftobiprole, tigecycline, oritavancin

CLSI guidelines for MRSA/MRS
Disc Diffusion Method using Cefoxitin –
Using MHA incubated in ambient air @ 33-35 degC for 24 hrs

using Cefoxitin 30 μg disc in reflected light
A} For S.aureus & S. lugdunensis

– R ≤ 21mm
– I – Nil
– S ≥ 22 mm
(changed in 2005 from 20mm to 21mm)
B} For CoNS-- R ≤ 24 mm ;S ≥ 25 mm,

WARNING:
• For oxacillin-resistant S. aureus and coagulase-
negative staphylococci (MRS), other β-lactam
agents, ie, penicillins, β-lactam/β-lactamase
inhibitor combinations, cephalosporins and
carbapenems, may appear active in vitro but are
not effective clinically.
• VRSA- Vancomycin Resistant Staphylococcus aureus –
MIC 32 µg/ml or more
• VISA - Vancomycin Intermediate Staphylococcus aureus -

MIC 8-16 µg/ml
• Mechanism – due to thickening of cell wall > VanA gene
• Treatment – same drugs given for MRSA except Vancomycin

& Teichoplanin.
Bacteriology
S.aureus is the MC agent for :
• Skin & soft tissue infection

• Botryomycosis
• Tropical pyomyositis – S.aureus , (acute bacterial myositis – GrA Streptococcus)
(Overall - S.aureus)
• Osteomyelitis, septic arthritis
• postoperative parotitis
• Paronychia
• Epidural abscess
• Surgical wound infection
4/4/2018 Bacteriology 35
• Lab D
Coagulase Negative Staph.
• Normal flora of skin
• They produce white non pigmented colonies &
can be distinguished from S. aureus by their
failure to produce coagulase and DNase
• Many stains of S. epidermidis are capable of
producing large no.of polysaccharide
glycocalyx known as slime (Biofilm
production)
• Coagulase Negative Staph. (particularly S.
epidermidis) are principal pathogens of;
Peritonitis in pts undergoing CAPD
Endocarditis with insertion of valvular
prosthesis
Ventricular shunt infections
• S. saprophyticus - Coagulase negative, causes

UTI in young sexually active female. It is
novobiocin resistant
NOVOBIOCIN SUSCEPTIBILITY
62. A patient admitted to an ICU is on central venous line for
the last one week. He is on ceftazidime and amikachin.
After 7 days of antibiotics he develops a spike of fever and
his blood culture is positive for gram positive cocci in chains,
which are catalase negative. Following this, vancomycin was
started but the culture remained positive for the same
organism even after 2 weeks of therapy. The most likely
organism causing infection is:
– Staphylococcus aurenus
– Viridans streptococci
– Enterococcus faecalis
– Coagulase negative Staphylococcus
•
• True regarding drug resistance of MRSA?
(AI 2011)
A. Penicillinase enzyme production
B. Due to change in penicillin binding
receptors
C. Plasmid mediated
D. Treated with amoxicillin + clavulanic acid
All are true about PBP except-
(AI 2010)
A. PBP is localized in outer cell wall
B. PBP is essential for cell wall synthesis
C. PBP acts as carboxy peptidase & transpeptidase
D. alteration in PBP – basis of MRSA

A pt has prosthetic valve replacement and he
develops endocarditis 8 months later.
Organism responsible is? (AIIMS nov 2010)
a.) Staphylococcus aureus
b.) Streptococcus viridans
c.) Staphylococcus epidermidis
d.) HACEK
The following are characteristic features of
staphylococcus food poisoning except-(AIIMS 04)
a. Optimum temperature for toxin production is 37°C
b. preformed toxin is responsible for intestinal
symptoms
c. Toxin can be destroyed by boiling for 30 minutes
d. Incubation period is 1-6 hours
True about methicillin-resistance staphylococcus
aureus (MRSA) – (PGI June 09)
a. Isoxazolyl penicillin is highly effective
b. All MRSA are multidrug resistance
c. Vancomycin is effective
d. MRSA are more virulent then sensitive strains
e. Resistance develop due to altered binding protein
45
All of the following statements are true regarding
Staphylococci except - (AIIMS Nov 04)
a. A majority of infections caused by coagulase- negative
Staphylococci are due to Staphylococcus epidermidis
b. Β-lactamase production in Staphylococci is under plasmid
control
c. Expression of methicillin resistance in Staphylococcus aureus
increases when it is incubated at 37°C on blood agar.
d. Methicillin resistance in Staphylococcus aureus is independent
of β-lactamase production
All of the following statements are true regarding
Staphylococci except - (AIIMS Nov 04)
a. A majority of infections caused by coagulase- negative
Staphylococci are due to Staphylococcus epidermidis
b. Β-lactamase production in Staphylococci is under plasmid
control
c. Expression of methicillin resistance in Staphylococcus aureus
increases when it is incubated at 37°C on blood agar.
d. Methicillin resistance in Staphylococcus aureus is independent
of β-lactamase production
47
A superantigen is a bacterial product that -
(AI 08)
a. Binds to B7 and CD28 costimulatory molecules
b.Binds to the beta chain of stimulating T cell activation
c. Binds to the CD4 + molecule causing T cell activation
d.Is presented by macrophages to a larger – than-
normal number of T helper CD4 + lymphocytes
48
NEET 2017
• Q-32. A cook prepares sandwiches for 10 people going for
picnic. Eight out of them develop severe gastroenteritis
within 4-6 hrs of consumption of the sandwiches. It is likely
that on investigations the cook is found to be the carrier of:
a) Salmonella Typhi
b) Vibrio cholerae
c) Entamoeba histolytica
d) Staphylococcus aureus
• Answer: Staphylococcus aureus
Explanation:
Organism——Incubation period
Staphylococcus aureus- 1-8 hours
Bacillus cereus- 1-8 hours
Streptococcus
Classification
of streptococci
Streptococcus pyogenes
Medically Important Streptococci
Blood agar:
•β hemolytic
colonies
–Addition of crystal
violet to Blood agar
inhibits the growth of
staphylococci, while
permitting the growth
of streptococci &
hence acts as
selective media for PNF medium is also selective
streptococcus for Streptococci (Polymyxin
(CVBA) Neomycin Fusidic acid)
Liquid media
Pike’s medium • Inhibiting the

growth of
staphylococci
Todd – •Used for luxuriant growth

Hewitt broth of streptococci in
grouping and typing
• Structural components of • Human tissue with which it
Strept pyogenes cross reacts
1. Caps. hyaluronic acid • a) Synovial fluid

2. Cell wall protein • b) Myocardium
3. Cell wall carbohydrate • c) Cardiac valves
4. Cytoplasmic membra • d) Vascular intima
5. Peptidoglycan • e) Skin antigens
➢Pharyngitis - Gp A Streptococci are the most common
Pathogenicity
bacterial cause of pharyngitis and primarily affects school
age children 5 to 15 yrs age
➢Scarlet fever – Any type of group A can produce
scarlet fever that produces erythrogenic toxin, dick test
+, Schultz Charlton reaction + (blanching of rash after
injection of antibodies).
➢Impetigo – This is caused predominantly by Strept
pyogenes; Also be caused by gp C& G & by S. aureus
Group A Streptococcus causing impetigo are frequently
nephritogenic leading to glomerulonephritis
➢Erysipelas - The sufferers are usually adults; Dick test
is negative early in disease
Non Suppurative
1. ARF - ~ 3% patients develop, 2-3 wks after
onset of acute pharyngitis, M types 1, 3, 5, 6, 12,
14, 17, 30, 32 & 41
2. AGN – can occur either after skin or throat
infection. Produced by gp A also group C may be
involved; Most cases of AGN occur about one
week after group A type 12 infections.
3. Erythema nodosum - Allergic reactions due to

the depositions of cell wall fragments
Determinants of pathogenicity: Cell wall component
Virulence factor Activity
Lipoteichoic acid Mediates adherence to fibronectin receptors on epithelial
cells
M protein Mediates adherence to epithelial cells, inhibits
phagocytosis
Hyaluronic acid Inhibits phagocytosis
capsule
Erythrogenic toxin 3types (SPE A, B and C). Type A&C bacteriophage
(SPE) coded, B toxin chromosomal
Superantigens
Dick test – (ID injection)- detect susceptible individuals
Schultz Charlton reaction (blanching of rash after injection
of antibodies)- diagnosis of scarlet fever
2) Exotoxins
(i) Erythrogenic toxins: (Streotococcal pyrogenic exotoxin)
• 3 antigenically distinct toxins A,B and C ( A & C phage mediated)
 Antigenic & neutralized by convalescent sera.

 Act directly on the hypothalamus & cause pyrogenecity
 Cause the characteristic rash of scarlet fever.
 These toxins are superantigens
Injected Into the skin of

Toxin susceptible child
Dick Test
Localized erythematous reaction appears

which reaches a peak at about 24 hours and the reac.
is caused by erthrogenic toxin
Virulence factor Activity
Streptolysin O Produced by group A, C and G streptococci.
Oxygen labile, Serum stable
Also Pneumolysin, tetanolysin, perfringolysin
Acts by binding to cholesterol in the cell membrane and
producing hole in it.
It is strongly antigenic and rise of ASO >200u, indicates
recent streptococcal infection
Streptolysin S Oxygen stable, Serum soluble
Hemolysis on the surface of an aerobic blood agar plate.
Not antigenic.
Streptokinase Fibrinolysin (activate plasminogen)
Rapid spread of infection by preventing the formation of fibrin
barrier.
a) Streptokinase (Fibrinolysin): Two types A & B
Streptokinase
Plasminogen Plasma
 Facilitates spread of streptococcal infection by preventing formation of
effective fibrin barrier around the lesion. (plastic Sx)
 This property is also used therapeutically in treatment of coronary
thrombosis.
b) Streptococcal deoxyribonuclease: streptodorunase Four types A, B, C & D

Degrade DNA, depolymerising the highly viscous DNA that accumulates in
thick pus derived from disintegrated polymorphonuclear leucocytes – anti
DNAase titers
 Enzyme preparation containing streptokinase and streptococcal
deoxyribonuclease used in clinically in liquifying localised collection of thick
exudates. e.g. pneumococcal empyema.
c) Nicotinamide adenine dinucleotidase (NADase):
d) Hyaluronidase:
 Favors spread of streptococcal lesion along intercellular
spaces.
e) Lipoproteinase (serum optical factor):
f) Other Extracellular products:

 Many members produce DPNase, neuraminidase,
phosphatase, esterase etc. are damaging to the tissues.
Group B Streptococci
S. agalactiae causes
 Early onset & late onset neonatal meningitis
 Early onset – first 5 days of neonates septicemia
& pneumonia
 Late onset between 2 to 4th week of life. Baby
acquires infection from hospital personnel
presents with menigitis
 Presumptive identification
1. CAMP +ve
2. Hippurate hydrolysis test +ve
3. pigment production
 S. Agalactiae grows on Mac media.
 Colonies are β-hemolytic
 Hydrolyze sodium hippurate and give positive CAMP test.
CAMP-Test
Group D Streptococci
• Gr D antigen- Teichoic acid
• They can be further divided into Enterococci & Nonenterococci
Enterococci
• Growth in presence of 6.5% NaCI , 40% bile, at pH 9.6 and at 450C , PYR +ve &
hydrolyze aesculin.
• resistant to penicillin, low level aminoglycosides & sulfonamides, cephalosporin,

Cotrimoxazole
• Combination therapy with penicillin & aminoglycoside is effective.
• Disease- UTI, endocarditis, intra abdominal infection, meningitis
• MC- Enterococcus fecalis & Enterococcus faecium ( more resistant)
• : Vancomycin
7/5/2018 resistant– due to Van gene
Bacteriology 67
• Viridans strept (oral) 
• S,sanguis, S.mitior, S.mitis,S. salivarus
(longest chains)S.milleri
–  hemolysis
– mouth commensals
– (S) to penicillin
– disease – dental caries, endocarditis
DIFFERENTIATING FEATURES
Test Enterococcus Group D
Streptococcus
Growth in presence
+ -
of 6.5% NaCl
PYR Test
+ -
Sensitivity to SXT
R S
(Trimethoprim-
Sulphamethoxazole)
Ability to grow at + -
45⁰C
True about MRSA all except
1. 30% are healthy carriers
2. Penicilinase production is chromosomally mediated
3. TSS is super Ag
4. Most common cause of infec. Is cross infection.

• Which of the following statements concerning
VISA is correct?
– Patient with VISA isolates need not be isolated.
– Minimum inhibitory concerning (MIC) for
vancomycin for VISA is at least 1.0 mcg/mL.
– VISAs have emerged because of the extended use
of vancomycin for methicillin resistant
Staphylococcus aureus (MRSAs).
– VISA isolates are infrequent, so surveillance at the
present time is not warranted.
– VISA isolates are usually methicillin susceptible.
NEET 17
• Q-35. The commonest organism causing
cellulitis is
a) Streptococcus pyogenes
b) Streptococcus fecalis
c) Streptococcus viridans
d) Micro-aerophilic streptococcus
NEET- 16
• Q-134. All are true about streptococcus except
a) M protein is responsible for production of
mucoid colonies
b) M protein is responsible for virulence
c) Mucoid colonies are virulent
d) No resistance to penicillin has been
reported
• Pneumococcus
Characteristics
 flame or lanceolate shaped
 capsulated gram positive cocci in pairs.
1. Bile sol
2. optochin (S)
3. ferments inulin
4. pathogenic to mice
• Infection caused by 1,2,3,4-8 -18 serotypes
– Non motile, non sporing gram +ve cocci
– Pair of flame shaped diplocci
– Capsulated – India Ink, Quellung Reaction
– Blood agar – zone of alpha haemolysis, further incubation
– Draughtsman colony - due to autolysis on blood agar
Biochemical Reaction:
S. pneumoniae are catalase and oxidase negative.
Bile solubility & autolysis:
Bile solubility test is positive.
Enzyme N-acetylmuramyl-L-alanine amidase

(autolysin) produced by pneumococcus solubilizes
the peptidoglycan cell wall.
This autolytic process is augmented by surface active

agents such as bile & bile salts.
• Bile soluble : activate autolytic enzymes
• Tube : 10% deoxycholate
• Plate : 2% deoxycholate
Differences
S. Pneumoniae S. viridans
Morphology Lanceolate Round/oval
Arrangement Pairs chains
Capsule + -
B/A Draughtsman Convex
colonies
Liquid medium Unform turbidity Granular
Bile solubility + -
Inulin fermentation + -
Optochin sensitivity S R
Pathogenicity P NP
Antigenic structure
• Capsular Ag – 90 types, SSS, important for

virulence, typing by Quellung reaction
• Somatic Ag – Carbohydrate Ag – teichoic acid
• Pneumolysin
• Autolysin – amidase
 There are three serotyps: Type 3 most virulent.
Quellung reaction: Done either in sputum/ culture.
Suspension of Mixed on slide with

encapsulated pneumococci a drop of specific antiserum
Capsule around pneumococci Loopful

reveals an apparent marked methylene blue
swelling sharply delineated
and refractile under the
microscope Quellung Test
Pathogenicity
• Commensal – 5-70% humans carry
S.pneumoneae in throat
• Pneumonia – In adults – type 1 – 8 , 18
• In Children – 6,14,19,23
• Lobar pneumonia : 10 – 50yrs
• Bronchopneumonia – young children, adults > 50years
• Meningitis
• Others – otitis media, sinusitis, peritonitis etc.
Therapy
• Rx – Penicillin.
• Penicillin resistance has emerged but this

resistance is not due to penicillinase enzyme .It
is chromosomally mediated and these are
altered penicillin binding protein
Vaccine
• 1. Polysaccharide vaccine (23 valent) –
• Not useful for <2yr as T independent
• C/I- lymph reticular malignancy, pregnancy
• Indication- spleenectomy, sickle cell patient, DM, chronic
heart/lung/renal/liver disease
• 2.Conjugated vaccine (7 valent)- can be given <2yr
• 'C' in C reactive protein stands for: (AI2011)
A. Capsular polysaccharide in Pneumococcus
B. Concanavalin-a
C. Calretinin
D. C carbohydrate Ag in Pneumococcus
• CRP:
• Acute phase reactant
• Betaglobulin produced by liver in response to

pneumococcal and some other bacterial infections
• Ca, inflamation.
• Ppt with CAg of pneumo but not an Ab
• Acute phase reactants – fibrinogen, transferrin, alfa1
antitripsin
60. Which one of the following bacteria is most
likely to be recovered from the cerebrospinal
fluid (CSF) of a newborn with meningitis
– Escherichia coli
– Streptococcus agalactiae
– Haemophilus influenzae type b
– Listeria monocytogenes
– Streptococcus pneumoniae
• Which of the following enzymes are used for
liquefaction of thick exudates as in empyema?
– Streptokinase & NADase
– Streptokinase & Streptodornase
– NADase & Hyaluronidase
– Streptokinase and N acetyl glucosaminidase
• All are true about Streptcoccus except-;
– In blood agar produce a hemolytic colony
– Crystal violet blood agar can be used as a selective
media for isolation of Streptococcus pyogenes.
– CAMP test can be used for presumptive
identification of Streptococcus agalactiae.
– Negative PYR test is a characteristic feature of
Streptococcus pyogenes.
• Pick the wrong statement from the following?
– Ludlam’s medium is the selective media for
Staphylococcus aureus.
– Staphylococcus aureus produces diffusible
pigment.
– Prompt phosphatase reaction helps to
differentiate St. aureus from St. epidermidis
– They produce black coloured colonies on
potassium tellurite agar
• A patient in an ICU , undergone CSF shunt
surgery. His blood culture shows growth of
grain positive cocci which are catalase positive
and coagulase negative. The most likely
etiological agent is:
– Staplylococcus aureus
– Staplylococcus epidermidis
– Streptococcus pyogenes
– Enterococcus faecalis.
Medically Important Gram-Positive
Bacilli
Three general groups:

1. Endospore-formers
Bacillus, Clostridium
2. Non-endospore-formers
Listeria, Erysipelothrix
3. Irregular shaped and staining properties
Corynebacterium, Proprionibacterium,
Mycobacterium, Actinomyces, Nocardia
CORYNEBACTERIUM
• GPB
• Club shaped, Chinese letter arrangement,
cuneiform, V & L forms/volutin granules/
polar bodies
• Stained with methylene blue / toludine blue -
metachromatic granules -Babe Earnst granules
• Inorganic polyphosphates
-CLUB SHAPED SWELLINGS (BOTH ENDS / ONE
END)
(Name: Coryne : CLUB)
Arrangement::-ANGLED PAIRS
PARALLEL ROWS
CHINESE LETTER PATTERN
(DIVISION OF BACILLI---ABRUPT
SNAP / BEND)
V / L PATTERN
Culture
• Loefflers serum slope
– Earliest growth (6-8hrs)
– Granules best developed
– Water of condensation can be used for animal
pathogenicity
• Selective medium – (used contaminated sites)
– Pottasium tellurite (.03 – 0.04% ) – black colonies due
to tellurite reduction.
– Tinsdales
– Hoyles medium
– Mcleod’s medium
BASED ON COLONY MORPHOLOGY
MITIS INTERMEDIUS GRAVIS
MILD INTERMEDIATE SEVERE
ENDEMIC EPIDEMIC ASSOCIATED

USUALLY
McLeod classification Gravis Intermedius Mitis
Colonies on PTA daisy head Frogs eggs colony poached egg
Fermentation of starch +ve -ve -ve
Toxigenic strains 100% 95-99% 80-85%
Epidemic Epidemic Endemic
complication Paralytic& Hemorrhagic Obstructive

Hemorrhagic
Hemolysis Variable non Hemolytic Hemolytic
Morphology Short, no granules Long barred, poor Long curved,

granules prominent granules
All 3 types are nitrate +ve

Belfanti (fourth biotypes )– Nitrate –ve variant of Mitis
97
• Diphtheria toxin +ve
• C. diphtheriae – (urease negative)
• C. ulcerans – (urease positive)
• C. pseudotuberculosis – (urease positive)
• Toxin is released locally but it causes systemic

S/S, toxaemia by absorption in circulation
• (Exo) Toxin A & B fragments
A inhibit elongation factor - inhibits protein
synthesis
Fragment B – for transportation of fragment A
inside cell
• Optimum iron concentration for toxin
production is 0.1mg per litre of the medium
toxin production stops 0.5 mg per litre of the
medium
• Toxigenicity due to - phage (tox phage)
• Almost all strains of gravis, 95%-99% of
intermedius& 80 – 85% mitis produce toxin
• The classic park Willian (PW8 ) strain of C.

diphtheriae is used as a source of toxin for
diphtheria toxoid (DPT vaccine)
Toxigenicity / Virulence test
• Toxigenicity is due to tox (ẞ) by Phage (lysogenic)
Conversion
• Tests for pathogenicity – Guinea pig or rabbitis used,
mice is resistant to diphtheria
• In vivo tests - Subcutaneous test, Intra dermal
• In vitro tests; - Elek gel precipitation test
• Difference between Diptheroids & C. diptheriae is

virulence tests
Susceptibility Test
• Shick test (toxin + antitoxin = neutralization)
• 0.2 ml (1/50 ml) toxin - I/d on ant. surface of

left forearm - while right forearm is control
with antitoxin - Results read after 24 – 48 Hrs
and 5-7 Days
• OBSOLETE
IMP POINTs
• Natural infection only in man.
• Pseudo membrane formation on tonsil, posterior pharyngeal wall.
• Source- carriers (Nasal carries are particularly dangerous - shed large no. of bacilli)
• Faucial diphtheria– MC, laryngeal – dangerous, requires tracheostomy
• 1st muscle involved – palatopharynges , recovery is the rule
• Ciliary but not papillary paralysis occurs – So blurred vision with intact light
reflex
• Neurological complication- occurs in 1st / 2nd week
• Systemic- myocarditis, peripheral neuropathy
• Cardiac damage is permanent but Nerve recovery usually occurs
• Death- due to circulatory failure
• Invasive infection – very rare, factors- pre existing cardiac anomaly, IV drug abuser,
cirrhosis.
TREATMENT
CLINICAL SUSPICION OF DIPTHERIA---- T/T- SHOULD NOT AWAIT
LAB CONFIRMATION
1ST STEP– DIPHTHERIA ( HYPERIMMUNE HORSE SERUM)
ANTITOXIN
•IMMEDIATE
•NO EFFECT ON TOXIN BOUND TO TISSUES
•NEUTRALISES ONLY CIRCULATING TOXIN
•DOSE DEPENDENT UPON SEVERITY--- 20,000 IU MILD
---100,000 IU SEVERE
C.diptheria sensitive to penicillin, Erythromycin, Rifampicin;

but antibiotics do not neutralize circulating toxin . Therefore
antibiotics are of value combined with antitoxin
Erythromycin active for treatment of carriers
OTHER MEDICALLY IMPORTANT CORYNEBACT./ NON DIPTHERIA
CORYNEBACTERIA
DIVERSE --STRICT AEROBE (environment)
GROUP FACULTATIVE COMMENSALS--SKIN
ANAEROBIC --M.m
2 GROUPS
INFECTION IN IMMUNOCOM OPPORTUNISTIC INFECTIONS

Petant INDIVIDUALS
C.Ulcerans C.jeikeium
C.pseudotuberculosis(C.ovis)
( preitz nocard bacilli) C.xerosis
C.Minutissimum C.pseudodiptheritic-
um(C.hoffmani)
C.bovis
C.parvum – immunomodulator
A patient presents with signs of pneumonia. The
bacterium obtained from sputum grows on sheep agar
producing green hemolysis. What test is used to
identify the type of organism? (AI2011)
A. Bile solubility
B. Bacitracin sensitivity
C. Coagulase test
D. Catalase test

A farmer presents with pustules showing gram
positive cocci, beta hemolysis, catalase
negative. To show it is group A streptococci,
which of the following test is done- (AI
07)
a. Bacitractin
b.Optochin
c. Novobiocin
d.Bile solubility
A patient admitted to an ICU is on central venous line for
the last one week. He is on ceftazidime and amikacin.
After 7 days of antibiotics he develops a spike of fever
and his blood culture is positive for gram positive cocci in
chains, which are catalase negative. Following this,
vancomycin was started but the culture remained
positive for the same organism even after 2 weeks of
therapy. The most likely organism causing infection is -
(AIIMS May 06, AIIMS Nov 06, AI 07)
a. Staphylococcus aureus
b. Viridans streptococci
c. Enterococcus faecalis
d. Coagulase negative Staphylococcus

A 65 years old man presenting with complaints of chest pain, fever,
cough with sputum. O/E of sputum pus cells with gram positive
cocci in pair present. Blood agar showed alfa hemolysis. How will
you differentiate this from other gram positive cocci?
(AIIMS Nov 09)
a. Bacitracin
b. Optochin sensitivity
c. Bile solubility
d. Coagulase positive
The sputum specimen of a 70 year old male was cultured on a 5% sheep
blood agar. The culture showed the presence of α-haemolytic colonies
next day. The further processing of this organism is most likely to yield-
(AIIMS Nov 05)
a. Gram positive cocci in short chains, catalase negative and bile resistant
b. Gram positive cocci in pairs, catalase negative and bile soluble
c. Gram positive cocci in clusters, catalase positive and coagulase +ve
d. Gram negative coccobacilli catalase positive and oxidase positive

Neonatal meningitis, beta haemolysis, bacitracin
resistant, CAMP positive-
a. S pyogenes
b. S agalactiae
c. S pneumoniae
d. S.aureus
(AI2010)

• False abt strep is – (AIIMS 2009)
a. Strptokinase produced by type A,C,G
b. Streptodorunase depolymerises DNA
c. AsO is active in reduced state
d. Pyrogenic exotoxin A is plasmid mediated
• Boy with skin ulcer on leg. Culture reveals beta hemolytic
Strept. Culture from school children with sore throat some
days back also revealed beta hemolytic Strept. What is the
characteristic that differentiate the two species affecting the
skin & throat? ( AIIMS nov 10)
• A. C carbohydrate Ag
• B. M protein
• C. emm protein
• D. Mec A gene
Mycobacterium
Acid fast Organism
• Mycobacteria (fungus bacteria) –
MTB/leprae/NTM
• Nocardia
• Rhodococcocos
• Spores
• Legionella mecdedei
• Parasites – Cryptosporidium, cyclospora,
Isospora, Tinea Scolex, Hooklets of Hydatid cysts
• M tuberculosis complex – include species
M. tuberculosis
M. bovis (bovine tubercle bacillius)
M. africanum (intermediate between M
tuberculosis & M bovis)
M. microti (vole tubercle bacillus)
• Mycobacterium – aerobic, non spore forming,

nonmotile
• Average generation time 14 – 15 hours
M.Tb vs M.bovis
Obligate Aerobes Microaerophilic
Eugonic, rough, buff, tough Dysgonic, white
Glycerol enhances growth Inhibits growth
Susceptible to Pyrazinamide Resistant
Resistant to thiophen 2 Sensitive

carboxylic acid TCH
NNN +ve (niacin, nitrate, -ve

neutral red)
Pathogenic to g. pig & NP to Pathogenic to both
Rabit
Pathogenicity
• Escape killing by macrophages & inhibits
phagolysozome fusion
• Virulence factor – cord factor, LAM, HSP
• Pulmonary TB – Primary TB – affect children,

sub pleural lower lobe lung – Ghon’s focus +
Hilar LN = primary complex
Lab methods
• Digestion & Decontamination procedures for
sputum
a) Petroff’s method (Na OH)
b) NALC – NaOH method (N acetyl lysine)
c) Zephiran trisodium phosphate
d) Oxalic acid method
Direct Smear Microscopy
• ZN / AF
• Kinyoun stain – cold
• Fluorescent staining (Rhodamine/ Auramine
O)
– 5 X 103 to 5 X 104 bacilli / ml of sputum is
required for detection by smear
– Culture detects 10 to 100 viable organism
CULTURE MEDIA
• Solid media
a) Egg based - Eg – 1. LJ, Petragnani, Dorset
b) Blood Based Media – e.g. Tarshis
c) Agar - media are transparent; colonies observed in
10 to 12 days in contrast to 18 to 24 days with
opaque egg based media. eg Middle brook 7H11,
Middle brook 7H10
• LIQUID MEDIA –
Middle brook 7H9
Dubus Tween albumin broths
• Rapid Culture Methods

BACTEC 460
MGIT
SEPTI Chek – biphasic media
ESP Culture system
MB/ BACT Alert
BATEC
• Radiometric detection system
• 14 C labeled substrates (palmitic acid)
• The amount of 14 CO2 is translated into growth
index.
• Average time to detect M. tuberculosis 9 to 14
days
• Detection time of positive cultures is decreased
to < 7 days for NTM
Other methods
• Serology Not much useful – Ag/Ab
• Quantiferon assay
• Molecular Methods
PCR - IS6110
DNA probes
Mantoux Reaction
• 0.1 ml PPD - i/d reaction read after 72 hours
• Induration >10 mm – positive
5-9 - doubtful
<4 – Negative
• Uses active infection in infants, prevalence of
infection
• false –ve – early/adv. TB, miliary TB,
measles ( immunity)
• false +ve - atypical Mycobacterial infection
Conventional Sensitivity Testing
1. Resistance Ratio
2. Absolute concentration
3. Proportion method
4. Radio metric method - based on proportion
method
5. Molecular methods
Newer DST Methods
• BACTEC 460
• MGIT
• PHAGE Plaque Assay
• Luciferase reporter phages
• PCR
• INNO-Lipa
• DNA sequencing
• DNA microarray
Sensitivity Testing
• Resistance Ratio
• Absolute concentration
• Proportion method – RNTCP recommended
• Radio metric method  based on proportion method
• Molecular methods
Drug Resistance- d/t mutation

• INH -Kat 6 gene, Inh A gene, ahpC
• R - rpoB gene ( RNA polymerase B)
• Z - Pnc A (Pyrazinamidase)
• E- emb A,B,C ( Arabinosyl transferase)
• S - ribosomal protein subunit 12(rpSL)
MRDTB- R/ INH& Rifampicin +/- R/other 1st line drug

XDRTB- MDRTB + R/quinolone+ R/aminoglycoside (amikacin/capreomycin/kanamycin)
• Neonatal TB
• Before delivery – If mother chest X-ray & sputum AFB +ve – mother given ATT
(HRE)
• After delivery –
• If mother chest X-ray & sputum AFB +ve then, mother to be given ATT(HRE) +
baby(INH for 9-12m) + screening of household contacts
• Separation from mother or with hold of breast feeding is not recommended
130
Radiometric BACTEC 460 TB method
• Based on generation of rCO2 from
14C labelled substrate palmitic acid .
• 14CO2 - is measured by the BACTEC

instrument and reported in terms of
growth index (GI).
• Detection in 5-10 days
• Used for diagnosis & drug

susceptibility testing
MGIT (Mycobacterial growth indicator tube)
• Automated system for growth detection by a

non-radioactive detection system.
• MGIT glass tube contains a modified

Middlebrook 7H9 broth with a oxygen
sensitive fluorescent sensor.
• Growing mycobacteria consume dissolved O2

and produce fluorescence when illuminated
by a UV lamp.
Vaccine
1. BCG (live attenuated vaccine)– 0-80%

efficacy – prevents systemic disease only
2. Recombinant Vaccines – under trial
3. DNA vaccines
DOTS under RNTCP
Category Type of
TB treatment regimens
of
patients
treatment Intensive phase Continuation phase
•New smear positive
New cases 2H3R3Z3E3 4H3R3
pulmonary
Tuberculosis
CAT-I (PTB)
• New smear negative
PTB
• New extra-pulmonary
TB.
Previously • Relapse,failure to 2S3H3R3Z3E3 +
5H3R3E3
treated respond or treatment
1H3R3Z3E3
after default
cases
• Re-treatment
CAT-II Others
Treatment Of DR-TB Cases
Category Type of TB treatment regimens
of
patients
Intensive phase Continuation phase
treatment
Cat IV MDR kanamycin, ofloxacin ofloxacin (levofloxacin),

(levofloxacin), ethionamide,
ethionamide, ethambutol and
pyrazinamide, cycloserine (18 months)
ethambutol and
cycloserine (6-9 months)
Cat V XDR Capreomycin, PAS, All drugs except
Moxifloxacin, High dose capreomycin (18
INH, Clofazimine, months)
Linezolid, Amoxy- Clarithromycin and
Clavulanic Acid (6-12 Thiacetazone as
months) substitute in intolerant
cases
Other Initiatives
• Ban of sero-diagnostic tests
• Availability of free quality assured anti-TB drugs
through local chemists
• New drugs: Bedaquiline, Delamanid (OPC-67683),
Linezolid, Sutezolid (PNU-100480), PA-824
• New vaccines- (Modified Vaccinia Ankara-MVA85)
• DOTS agent are paid incentive of Rs. 700 for NSP
case and Rs. 5000 for MDR-TB case.
NTM/MOTT/ Atypical
Mycobacteria
• NTM are distinguished from tubercle bacilli by
their resistance to both paranitrobenzoic acid
& TCH
• Aryl sulphatase positive
• Resistant to Anti tubercular drugs
• Strong Catalase +ve
Runyon group
I Photochromogen: M kansasii, M marinum,
M simiae (KaSM)
II Scotochromogens: M. scrofulaceum, M.
Suzulgai (photo at 25C), M. gordonae
(SSS-G)
III Non chromogens: MAC, M xenopi M.
Ulcerans
IV Rapid growers: M chelonei, M fortuitum
Diseases caused by NTM
Disease Causative Agent

Johne’s diseases M. Paratuberculosis
Lymphadenopathy M scrofulaceum, M avium intracellulare
M szulgai
Pulmonary disease M avium intracellulare, M kansasii
M xenopi, M simiae, M fortuitum
Skin lesions: Buruli ulcer M lucerans
Swimming pool granuloma M marinum
Post-injection / post M chelonei
traumatic abscesses M fortuitum
Disseminated disease M avium intracellular

Organisms requiring incubation at low
temperature (25C to 33C)
• M marinum
• M ulcerans
• M chelonae
• M haemophilum
Microaerophilic – M. bovis
Saprophytic – M.smegmatis, M. phlei
• MYCOBACTERIUM LEPRAE
• Non cultivable , generation time- 20days
• Grows well in cooler area (skin, testes, peripheral nerve, anterior eye)
• 5% H2 SO4 Acid Fast
• Intracellular-
• as parallel cigar bundles of bacilli bound with lipid like glia (globi) present inside
foamy macrophage(Virchow’s leprae cell)
• Bacteriological index – total no of leprae bacilli
• Morphological index-uniformly stained
• MI is more meaningful for monitoring t/t

Epidemiology:
• Transmission- nasal secretion, IP- 2-5year
• Not highly communicable, intimate & prolong contact necessary
• Most affected area- Southeast Asia& Brazil In India(MC-Bihar>Orissa>UP)
Animal model-
• foot pad of mouse(Shepard model )
• Nine banded armadillo is highly susceptible to leprosy, due to low body temperature.
Classification
• Ridley Jopling classification TT, BT, BB, BL, LL
• Madrid classification – LL, TT, borderline/dimorphous, indeterminate (early unstable type)
• Indian classification – Madrid + pure neuritic type

• Lepra Reaction Type I- • TypeII reaction-
• HSN IV, • HSN III,
• a/w borderline • a/w BL,LL
• down grading reaction • Usually follows sulfonamide
• upgrading reaction therapy, but may precede t/t
• MC feature- edema • MC- crop of painful erythematous
• ↑ Tcell (γδ TCR) papule
• MC nerve- Ulnar N • Central cytokine involves- TNFα
• t/t- DOC- glucocorticoid> • T/T- DOC- thalidomide,
clofazemine, thallidomide glucocorticoid, clofazemine &

antipyretics
Lepromin reaction  0.1ml Lepromin given i/d
• (1) Early/Fernandez - 2-3days- Induration like tuberculin (DTH)-
indicates Inf. in past(not useful)
• (2) Late/Mitsuda - 3-4 weeks  nodule, necrosis, ulcer
Measure of CMI induced by injected Lepromin
(doesn’t say about the past exposure or not used for diagnosis)
Uses – 1.classify lesions of leprosy +ve in TT
2. Assess prognosis +ve indicates good prognosis
3.Assess resistance to leprosy -ve means more prone to leprosy

Lab diagnosis:
• Sample- minimum 4 skin (slit skin smear from edge , NOT centre)
(butttock/forehead/chin/cheek)+ ear lobule+ nasal mucosa –
• grading of smear is done based on MI.
• Acid fast staining with 5% sulfuric acid
• Mouse food pad inoculation
• Ab to PGL1
Treatment-
• Paucibacillary- (I ,TT, BT)- Rifampicin(monthly)+ dapsone daily – for 6m
• Multibacillary- (BB,BL, LL)- Rifampicin(monthly)+ dapsone daily +clofazemine
daily –till 1 years or smear -ve
• Single lesion – (ROM)- rifampicin+ ofloxacin+ minocycline – single dose
61. True regarding pathogenecity of
Mycobacteria species?
– M.tuberculosis is more pathogenic than M.bovis
to the humans
– M.Kansasii can cause a disease indistinguishable
from tuberculosis
– M.africanum infection is acquired from the
environmental source
– M.marinum is responsible for tubercular
lymphadenopathy
a. Sterile pyuria is present in? (AI 2011)
A. Renal tuberculosis
B. Chronic hydronephrosis
C. Wilm's tumour
D. Neuroblastoma

Which of the following is true regarding globi in
a patient with lepromatous leprosy - (AI 02)
a. Consists of lipid laden macrophages
b.Consists of macrophages filled with AFB
c. Consists of neutrophils filled with bacteria
d.Consists of activated lymphocytes

The following test is not used for diagnosis of
leprosy - (AIIMS 06)
a.Lepromin test
b.Slit skin smear
c. Fine needle aspiration cytology
d.Skin biopsy

Collection of urine sample of a patient of TB
/kidney (AIIMS June 2000)
a.24 hrs urine
b.12 hrs urine
c. In early morning
d.Any time

Rapid examination of Tubercle bacilli is possible
with- (AI 96)
a.Ziehl –Neelsen stain
b.Kin young stain
c. Auramine –Rhodamine stain
d.Giemsa stain

Anti-tubercular drug susceptibility can be done
by all of the following methods, except-
(AIIMS May 04)
a.Resistance ratio method
b.Disc diffusion method
c. Molecular method
d.Radiometric broth method

Scotochromogens are - (PGI Dec
08)
a. Mycobacterium gordonae
b.Mycobacterium marinum
c. Mycobacterium intracellulare
d.Mycobacterium avium
e.Mycobacterium kansaii

True about Mantoux test- (PGI June 03)
a. < 5 cm always +ve
b.Usually –ve after treatment
c. Positive reaction in children < 2 yrs is not
important than adult
d.Usually read after 48-72 hours
e.False +ve in post measles state

True regarding mycobacterium tuberculosis is-
(PGI June 02)
a. Produces visible colonies in 1 weeks time on
Lowenstein – Jensen media
b.Decolorized by 20% sulfuric acid
c. Facultative aerobe
d.Niacin positive

All the following are true of Tuberculosis except
- (AI 94)
a. For sputum to be positive, bacilli should be > 104 /ml
b. Niacin test differentiates M.tuberculosis and M.bovis
c. Pathogenicity to rabbits differentiates M.tuberculosis and
M.bovis
d. Culture techniques have low sensitivity

• Basanti, 29 year aged female from Bihar present with active
TB . She delivers baby. All of the following are indicated
except- (AI 01)
a. Adiminster INH to baby
b. Withhold breast feeding
c. Give ATT to mother for 2 years
d. Ask mother to ensure proper disposal of sputum
161
ACTINOMYCETES
• gram positive bacteria varying from coccoid & pleomorphic
forms to branched filaments.
• Human pathogenic actinomycetes :
• Actinomyces (3) Streptomyces
• Nocardia (4) Actinomadura
• Actinomyes is non acid fast and anaerobic

or micro aerophilic
• Nocardia – Aerobe & acid fast (1%)
• Streptomyces & Actinomadura are aerobes & non acid fast
• Actinomyces  Causes Actinomycosis MC caused by A israelii

Gram Positive Filamentous Bacteria
GENUS OXYGEN GRANULE ACIDFAST
Mycobacteria aerobe no yes
Actinomyces ANAEROBE YES no

COMMENSAL
Nocardia Aerobe sometime PARTIALLY
SAPROBE
Actinomadura aerobe sometime no
Streptomyces aerobe YES no

• All these species are commensals of the mouth,GIT
and F Genito UT therefore cause endogenous
infections.
• Crtical step- breach in mucosal barrier
• Orocervical – Lumpy jaw (angle of jaw MC site)
• Abdominal abcess
• Brain abcess
• Ac/chr osteomylitis
• Endometritis – ass with IUCD
• Microscopy  sulphur granules are crushed
between slides & stained with Grams & ZN
• Granules consist of gram positive bacilli &

surrounded by peripheral zone of swollen radiating
club shaped structures presenting sunray
appearance.
• Clubs are Acid fast and are of host origin

• Culture BHI agar ,thioglyco.broth
• Incubation 14 days
• A israelli col.- molar teeth
• Nocardia strictly aerobic

• Unlike Actinomyces – Nocardia are environmental saprophytes
• Nocardia – grow on Sabourads dextrose agar, brain heart infusion

agar
• for isolation of Nocardia from soil, paraffin bait technique.

• Stains: AFB, Alcian blue, mucicarmine Kinyons,
Colour of the granules of various causative agents of
Actinomycetoma
Causative agent Colour of the grain
Actinomadura madurae White-yellow
Actinomadura pelliteiri Red
Nocardia asteroides White -yellow
Nocardia braseliensis White
Streptomyces Yellow
somaliences
• What additional tissues are frequently
involved with diphtheria besides the “bull
neck” and the pseudomembranous
pharyngitis?
– Skin (cutaneous diphtheria)
– Kidneys
– Heart and nerves
– Liver and kidneys
– Ears and sinuses
48. Virulence of Corynebacterium diphtheria is
associated in Lab by
– Metachromatic granules
– Elecks gel precipitation
– Glycogen production
– Daisy head colony as growth
47. True statement regarding diphtheria toxin is
– Toxin produced by all corynebacteria
– Positive Schick test suggest immunity
– Toxin production is influenced by critcal conc of
iron
– It is an Endotoxin
46. Which of the following statements about
diphtheria are untrue
– Immunity to diphtheria depends circulating
antitoxin
– Humans are the only hosts for the organisms
– Toxin is produced by only strains lysogenic for
bacteriophages carrying the tox gene
– Fragment A of the toxin is responsible for
attachment to host cells
• False about C.diphtheriae is? (AI2011)
A. Toxin production is chromosome mediated

B. Iron is required for toxin production
C. Toxin inhibits protein synthesis
D. Non toxigenic strain also causes infection

Clinical diphtheria is caused by- (PGI June 09)
a. Corynebacterium diphtheriae
b.Corynebacterium parvum
c. C.ulcerans
d.Streptococcus pyogenes
e.C.Pseudodiphthericum

In a completely and adequately immunized child against
Diphtheria, the throat swab was collected. It showed
the presence of Corynaebacterium diphtheria like
organisms on Albert staining. These orgainisms can
have one of the following properties on further
laboratory processing- (AIIMS Nov
04)
a. It can grow on Potassium tellurite medium
b. It would show a positive Elek’s gel precipitation test
c. It can be pathogenic to experimental guinea pigs
d. It can produce cytotoxicity in tissue cultures

Positive shick’s test indicates that person is- (AI
02, AIIMS Nov 07)
a. Immune to diphtheria
b.Hypersensitive to diphtheria
c. Susceptiable to diphtheria
d.Carrier of diphtheria

A child presents with a white patch over the
tonsils, diagnosis is earliest made by culture in
- (AI 01)
a. Loeffler medium
b.L.J.media
c. Blood agar
d.Tellurite medium

Clinical diphtheria is caused by- (PGI June
09)
a. Corynebacterium diphtheriae
b.Corynebacterium parvum
c. C.ulcerans
d.Streptococcus pyogenes
e.C.Pseudodiphthericum

Bacillus
• family Bacillaceae
• aerobic gram positive bacilli
• in human and animals  2 species
• (non motile) B.anthracis – highly pathogenic

• B .cereus (motile)- FP
• B.anthracis historic relevance

• first bacterial vaccine
• first pathogenic bacteria seen under microscope
• first communicable disease transmitted by infected blood
• first to be isolated in pure culture
• Presumptive diagnosis : MC fadyean reaction –
capsule seen when stained with polychrome
methylene blue in blood film
• Spores found in soil or culture ,never in animal

body
• In culture : medusa head appearance – chain of

bacilli,
• Grams: bamboo stick appearance
• Gelatin stab: inverted fir tree appearance
• PLET media- poly, lyso, edta, thallus acetate
Medusa head colonies
On sheepblood agar the colonies are non-haemolytic 2-3mm in diameter,

irregular raised, opaque and greyish white with "frosted glass" appreance. The edge of the
colonies are curled or fringed having a medusa head appearance
Anthrax
• caused by B.anthracis
• It is a zoonosis (cattle and sheep)
• Man acquire infection through - small cuts or

skin abrasion
• spore inhalation
• ingestion of meat (rarely)

Types of anthrax
Cutaneous anthrax - 95% of human anthrax, black eschar,
malignant pustule seen in farmer’s veterinary surgeons, persons
handling animal carcasses hides, Hide porter’s disease,
duckering 2% formaldehyde
Pulmonary anthrax - wool sorter’s disease (inhalation of spores)
Intestinal – violent enteritis, bloody diarrhoea

Enzootic, epizootic, zoonotic disease
Bioterrorism - recently used by Afghanistan

spores enclosed in paper envelopes were mailed - spore
inhalation
Virulent factors
• Capsule – inhibits phagocytosis

• Capsule- polyglutamate- plasmid mediated,
• Toxin 3 fractions- EF, protective Ag(binding), lethal factor
(cell death)
• Toxin plasmid mediated
live attenuated spore vaccine (Sterne, Mazucchi)
cutaneous anthrax  95% of human anthrax may resolve

spontaneously
Anthrax bacilli Anthracoid bacilli
Non motile Motile
Capsulated Noncapsulated
Long chain Short chain
Medusa head colony Absent
Non –Hemolytic colony on blood hemolytic colony
agar
Broth- turbidity absent Usually turbid
Salicin not fermented Salicin fermented
Susceptible to gamma phage, Resistant
penicillin, chloral hydrate
Inverted fir tree appearance & slow Rapid gelatin liquefaction
gelatin liquefaction
No growth at 45c Usually grows
Pathogenic to lab animal Non pathogenic
Bacillus cereus
• motile
• lacks capsule
• food poisoning
• Toxin . emetic toxin – heat stable

• symptoms after 1-6 hours of ingestion  preformed
toxincaused by eating boiled or fried rice –
• V,abdo cramps no fever
• Entertoxin – heat labile
• symptoms develop after 10-12 hours
• Culture media- MYPA
Listeria
• Gram +ve coccobacilli
• Tumbling motility (motile at 250 C)

• Grow at lower temp 2 – 43 0 C (cold enrichment)
• It can grow in refrigerated food & can tolerate preserving agents

• Milk born/ nosocomial/ preg/ neonate
Infections
• Pregnancy & Neonatal infections – Before 20 wks is rare. May
lead to abortion, still birth, early onset neonatal disease
• Neonate: septicemic illness appear with in 2 days after delivery

(granulomatosis infantiseptica) – disseminated infec. lung liver
abcess
• Late onset – mainly meningitic,
• D/T hospital cross infection & symptoms appear 5 days or more
after birth
• I/C – menengitis, sepsis, endocarditis,
• Lab diagnosis  Catalase +ve

• Beta haemolysis – produces haemolysin called listerolysin
• Specimens inoculated on BA, CA, PALCAM agar

• Anton test
• TT- Ampi/ cotrimox in allergic pt

• Resistant to cold, heat, salt, pH
extremes and bile
• Pathogenesis – survival &

multiplication in cells
• Avoids humoral immune system

• CMI plays role
– host cell actin polymerization
– listeriopods
• Bacteria uses host cell actin
to spread - Listeria, Shigella,
Rickettsia
• Diphtheria toxin:
– Acts catalytically
– Releases incomplete polypeptide chain from
ribosome
– Activates translocase
– Prevents release factor from recognition terminal
signal
– Attacks RNA of large subunit
A 35-year-old fisherman presents with a pruritic
nonsupporative lesion on the left index finger where
he had nicked with knife while gutting fish. He has
severe throbbing pain in his finger. The lesion has
spread about ½ cm around the initial nick and the
center has a discolored area. A scraping from the
periphery yields nonhemolytic, Gram-positive bacilli.
The organism isolated is most likely to be:
Diferertial: Staphylococcus aureus - Erysepelas

Erysipelothrix rhusiopathiae
• Clinical significance
– Primarily a pathogen of swine, turkeys, and fresh water fish.
– In man, the disease called erysipeloid is the most common

form.
– It is an occupation associated disease in which a reddish-

blue, edematous lesion at the site of inoculation, primarily
following trauma to the hands.
– Occasionally the organism disseminates to cause septicemia,

endocarditis, and arthritis.
Erysipeloid
Whale finger
• Characteristic of Bacillus cereus food
poisoning is (AIIMS 2010)
a. presence of Fever
b. presence of Pain abdomen
c. Absence of Vomiting
d. absence of Diarrhea

• Malignant pustule (AIIMS Nov 2010)
a. Sq cell Ca
b. Malignant melanoma
c. Anthrax
d. Infected rolent ulcer

A man, after skinning a dead animal, developed a pustule
on his hand. A smear prepared from the lesion showed
the presence of Gram positive bacilli in long chains
which were positive for McFayden’s reaction. The most
likely etiological agent is- (AI 04)
a. Clostridium tetani
b. Listeria monocytogenes
c. Bacillus anthracis
d. Actinomyces sp

Anthrax bacilli differs from anthracoid bacilli by
being - (PGMEE 06)
a. Non- capsulated
b.Strict aerobe
c. Non-motile
d.Haemolytic colonies on blood agar

True regarding anthrax is all except- (AIIMS
Dec 97)
a. Caused by insect bite
b.Caused by rubbing of skin
c. Cutaneous type is rare nowadays
d.Pulmonary infection occurs by inhalation

What of the following is true regarding anthrax-?
A .M’Fadyean reaction shows capsule

C .Less than 100 spore’s pulmonary infection
D .Gram stain shows organism with bulging spores
E .Sputum microscopy helps in diagnoses
F .Plasmid is responsible for toxin production
G .Cutaneous anthrax generally resolve spontaneously
H .Capsular polypeptide aids virulence by inhibiting phogocytosis
I .Toxin is a complex of two fractions
All of the following are true about Listeria except
-
(AI 02)
a.Transmitted by contaminated milk
b.Gram negative bacteria
c. Cause abortion in pregnancy
d.Causes meningitis in neonates

NEET 17
• Q-150. Which of the following is true

regarding globi in a patient with lepromatous
leprosy?
a) Consists of lipid laden macrophages
b) Consists of macrophages filled with AFB
c) Consists of neutrophils filled with bacteria
d) Consists of activated lymphocytes
• Gram Negative Cocci
N.MENINGITIDIS
MORPHOLOGY
• oval , gram negative diplococci
• opposing edges flat/ concave
• Intracellular (inside PMNLS) in

smears from clinical specimens
• Capsule present in fresh isolates

• pili present-mediate attachment
to host cells (virulence factor)
BIOCHEMICAL CHARACTERISTICS
• Oxidase Positive
• Acid formation is weak and oxidative
• Carbohydrate Utilisation Tests
GLUCOSE MALTOSE LACTOSE SUCROSE

N.GONO + - - -
N.MENIG + + - -
• Antigenic Structure  possess polysaccharide
capsule & divided into 13 sero groups A, B, C, D, X, Y,
Z, 29E W 135, H, I,K, & L
• Group D  Not found naturally
• Most meningococcal disease  A B & C
• Small proportion  Y & W135
• X Z & 29 E  rarely associated with some form of

immuno deficiency
• H, I, K, &L  from carrier & have not been associated

with disease
Determinants of pathogenicity
1. Capsular polysaccharide  inhibits phagocytosis
2. Endotoxin  As organism invade & multiply they release large

quantities of lipopolysaccharides & cause vascular necrosis
& produce generalized Shwartzman reactions & are
involved in pathogenesis of Water House – Friderichsen
Syndrome
3. IgA 1 protease  IgA1 protease cleaves subclass IgA 1 &

inactivates it.
4. Pili  allow to adhere & antiphagocytic
• Pathogenicity
• Meningococci are normally carried in nasopharynx of
5-10% of healthy individual
• During epidemic  carrier rates range from 20-90%
• complication B/L adrenal hemorrhage
Waterhouse friderichsen syndrome.
• Treatment  Cefotaxime & ceftriaxone  also
effective for H. influenza & Strept pneumoniae .
important to give Rifampicin or ciprofloxacin to
eradicate N. meningitidis from Nasopharynx. Vaccine
– Gp specific  A,C,Y ,W-135 immunity lasts for 3
years
– No vaccine for GpB: capsule is poorly immunogenic
N. gonorrhoea
• Adjacent sides are concave – kidney shaped.
• Urethritis prostatitis, epidymitis- males
• Females less severe., no vaginal involvement, cervicitis
• Water can perineum
• Fish haugh curtis syndrome
• – more difficult to grow than meningococci
• Kellogg divided gonococci into 4 types based on pili
• T1-T4
• T1 T2  Numerous pili P+ P++, and are virulent
• T3 & T4 – Non pillated or P- , avirulent, forms smooth

suspension
Selective Media
 Thayer-martin Medium
 Modified T.M .Medium
 Martin-lewis Medium
 New York City Medium
• Transport media (Non-nutritive )

 Stuarts medium
 Amies medium
 Cary-Blair medium
• Transport cum growth media (Nutritive)

 Transport plus selective growth medium plus carbon
dioxide environment
 Transgrow / Jembec
VIRULENCE FACTORS
• Pili
– mediate attachment of gonococci to host cell
– anti-phagocytic
– piliated strains are virulent
• outer membrane proteins

– P I & P III
• involved in adherence of bact. to host cell
• Form transmembrane porins involved in
exchange of molecules across outer membrane
• Babies born to infected women
• 
• ophthalmia neonatorum (in first week of life)
• (30% due to N. gonorrhoea)
• Treatment Chlamydia (2nd and subsequent week)
• 1% of AgNO3
• or
• 1% tetracycline
• or
• 0.5% erythromycin
Gram Negative Bacilli
• True statements is:
• A Solid media are enrichment media
• B Nutrient broth is a basal media
• C Agar adds nutrients to media
• D Chocolate agar is a selective media
General characters of family
Enterobacteriaceae
• Gram –ve bacilli
• Non-acid fast & non-sporing
• Aerobic / facultative anaerobic
• Grow readily on ordinary media
• Motile by peritrichate flagella / non-motile
• Ferment glucose with acid or acid & gas
• Reduce nitrates to nitrites
• Catalase +ve
• Oxidase –ve
ESCHERICHIA COLI
• GNR, non-capsulated, motile by peritricyhous, 80% strains
Fimbriae present, some strains capsulated
•
• strains hemolytic on Blood agar, Lf on Macconkey’s agar
• Inhibited on DCA, Ss agar, W& B agar
• Indicator media for e Coli- 0157:H7 – S.mac conkey agar

- Rainbow agar
• Biochemical reactions
• IMViC: ++--
• PPA, Urease, H2S Liquefaction, KCN: Negative
• Glucose, Mannitol, maltose, sucrose, Lactose: AG

Antigenic structure
• O: Somatic antigen, Heat stable, LPS, 174, cross reactions
• H: Flagellar antigen, thermolabile, 75, Few cross reactions
• K: Capsular antigen, 103
• F: Fimbrial antigen, thermolabile, Help in adhesion
• Normal colon strains: “early “O groups (1, 2, 3, 4 etc);

enteropathogenic strains, “later” O groups ( 26,55,86,111 etc.)
• Virulence Factors
Surface antigens
• “O” antigen (LPS): endotoxic; inhibits phagocytosis
• K antigen; protects from phagocytosis
• Fimbriae:
» Mannose resistant
» CFA (I-IV) colonizing) factor antigens ) in ETEC
» “P” Fimbriae in uropathogenic E.coli
Toxins
• Hemolysin: exact role is not known
• Enterotoxins :
– LT (Labile)
– ST (Stable)
– VT/SLT (Shiga like toxin or verocytotoxin)
LT:
Resemles cholera toxin in structure (A1 B5), ag properties and mode of
action (Gm1 ganglioside receptors-activates Adenylate cyclase –cAMP –
fluid accumulation)
• LT1 & LT2
• Powerful Ligated rabbit ileal loop test (18hrs.), steroid

production in Y1 mouse adrenal cell culture, morphological
changes in Chinese hamster ovary cells, latex Agglutination,
ELISA.
• ST: (Plasmid coded)
– Low molecular weight

– Poorly antigenic
– 2 types : ST-1 & ST-II
– ST-I: activation of cGMP
• Ligated rabbit ileal loop test (6hrs.) intra gastric in sucking mouse
– ST-B: mechanism of action not known
• VT:
– E.coli O157: H7
– VT1, VT2
– VT: Identical to shiga toxin, phage encoded, Cytotoxic to vero cells,
– VT is not neutralized by shiga antitoxin
Methods for detecting ETEC enterotoxins
In vivo tests LT ST
Ligated rabbit
ileal loop
-6hrs ± +
-18hrs + -
Infant mouse - +
intra-gastric
Adult rabbit + -
skin
In – vitro tests LT ST
Steroid production + _
in Y1 mouse
adrenal cell culture
Morphological + -
changes in CHO
cells
RIA + -
ELISA + +
BIKEN + -
CLINICAL INFECTIONS PRODUCED BY E.coli:
• Pyogenic infections
• Septicemia
• UTI:
– Most common cause
– “Early” O groups 1, 2, 4, 6, 7, 18,75
– Ascending infection: pyelonephritis, K antigen
– Kass’ concept of “significant bacteriuira”
– Semi quantitative culture
– P fimbria are uropathogenic, afimbrial adhesisns
Diagnosis-
• If delay – urine can be refrigerated or stored with 1.8% boric acid
Screening –
• Griess nitrite test, Catalase, triphenyl tetrazolium phenyl test,
• leucocytes esterase test, Wet mount (↑leucocytes)

EPEC
 Diarrhoea in infants and children, sporadic diarrhoea in adults
 O26, 55,86,111,114,125,126,127,128,142
 Pathogenicity : adherence to intestinal mucosa –disruption to

brush border microvilli –attaching effacing lesions.
 3 patterns of adhesion; localized, aggregative and diffuse

Diagnosis
• Fluorescent Actin staining method
• HEP-2 or Hela cell culture to demonstrate localized adherence
• Molecular diagnosis
• ETEC:
– O6,8,15,25,27,63,78,115,148,153, 159,167
– Pathogenesis : LT, ST
- Adhesive Fimbrial proteins (CFA I, II, III, IV)
• Diagnosis
– Typing
– Acute watery diarrhoea in infants and adults: traveler’s
diarrhea
– Demonstration of toxins in tissue culture by latex
agglutination, ELISA (LT) & RIA (ST), animal models
EIEC:
– Dysentery like disease in all ages
– Disease similar to shigellosis (flex)
– O28, 112,124,136,143,144,152,164
– Pathogenesis: Epithelial cell invasion by virulence marker Ag(VMA)
Diagnosis
– Typing
– Atypical biochemistry (biochemically similar to Shigella fl..)
– Plasmid probes
– Hela & Hep2 cell invasion assay
– Sereny’s test
Uninoculated HEp-2 cells
HEp-2 cells invaded by EIEC.
Sereny’s test - keratoconjunctivitis
EHEC (VTEC)
– O157:H7
– Bloody diarrhoea in all ages: HC: HUS
– Pathogenesis: VT1 and or VT2,,
Capillary microangiopathy
• Diagnosis
– Atypical biochemical (β-glucurronidase negative, donot ferment
sorbitol & rhamonose)
– VT1 & VT2 probes
– Demonstration of VT1 & VT2 in Vero cell lines
EAEC:
– Persistent diarrhoea especially in developing countries
– Most are “O” un-type able but “H” type able
– Aggregated in a “Stacked Brick” formation on Hep2 cell lines
– EAST 1 (enter aggregative heat stable enterotoxins)
43. All are true about gonococci except
– G +ve diplococci
– Causes uretheritis stricture
– Casuses salphingitis
– None of the above
10. Characteristics of outer membrane of
meningococci are all except
– Contains lipo oligosaccharide

– Induces tumor necrosis factor
– Induces interlukein III
– May cause complications like shock &
hemorrhage
– Lipopolysaccharide endoxin is analogous to lipo
oligosaccharide
11. Neisseria bacterimia is favoured by
complement deficiency of
– C1 to C2
– C1 to C3
– C1 to C4
– C5 to C8
– C1 to C6
• False regarding N. gonorrhoea
– Exclusively human pathogen
– Acute urethritis most common manifestation in
man
– Tenosynovitis is complication
– Uniformly sensitive to penicillin
•
5, Following are true about gonococci &
meningococci except
– Both are oxidase positive
– Ferment both glucose & maltose
– Both have asymptomatic carriers
– Only meningococci have polysaccharide capsules
42. Wrong about Ureaplasma urealyticum
– T strain
– Colonies are 5 to 25u
– Does not require cholesterol for growth
– Hydrolyse urea
– Susceptible to thallium acetate
• T strain- tiny col of mycoplasma

6. Best site to obtain swab in asymptomatic
gonorrhoea is
– Endocervix
– Lateral vaginal wall
– Urethra
– Posterior fornix
• Prophylaxis against meningococcal
meningitis is done with
– Rifampicin
– Penicillin
– Ciprofloxacin
– Chloramphenicol
KLEBSIELLA
• CAPSULE; LF, NM, produce mucoid colonies
• Urease +ve
• K. pneumoniae, ozaenae, rhinoscleromatis
• FREIDLANDER PNEUMONIA  Multiple abscess formation with

severe bronchopneumonia and Chronic destructive lesions
• K. pneumoniae – Pneuminia, pyogenic infections, UTI, septicemia
• K., ozaenae- atrophic rhinitis, ozenae
• K. rhinoscleromatis – garanulaomatus ds – rhinoscleroma

• PROTEUS, MORGANELLA, PROVIDENTIA
• Motile, NLF, swarming growth,
• Diene’s phenomenon – to know the relatedness between different
strains
• 4 Spp.
• Pr. vulgaris Urease+ve
• Pr. mirabilis Most Common PPA+ve
• Pr. penneri
• Pr. myxofaciens H2S+ve
• Non motile - OX 19, OX2,
OXk
•

• (Weil felix) Pr vulg. 01,02
Pr mirab 03
• Fishy seminal odour
• Swarming inhibited on MAC, CLED
• Swarming inhibited – by 6% agar, chloral hydrate, boric acid, alcohol,
surface active agents
• Organism which swarm- Proteus, V
parahemolyticus, V alginolyticus, Serratia
• Clostridium tetani,
• Not true regarding E.coli that causes HUS is
– Vero & Hela cells cytotoxicity
– Produces shiga like toxin
– Sorbitol fermentation
– Positive Sereny Test
• Biken test is ____ reaction used for E. coli.
– Precipitation
– Agglutination
– Toxin antitoxin assay
– CFT
• The mechanism of action of EPEC is
– Adherence to enterocytes
– Stimulates adenylcyclase
– Produce secretary diarrhoea
– None of the above
• A young lady presents with fever, dysuria and pain abdomen.
Uncomplicated acute cystitis was diagnosed. Which among
the following is not true? (AI 2011)
A. Nitrate test positive
B. E.coli count was < 103
C. 1 pus cell per 7 fields
D. 1 bacilli per field

• A patient is kept on ceftriaxone and amikacin, ESBL klebsiella
infection. What will you do next?
(AIIMS nov 2010)

a. Continue with same antibiotic but in higher dose
b. Change ceftriaxone and add ceftazidime
c. Start imipenem in place of ceftriaxone
d. Remove Amikacin

b lactamase
AMBLER classification-
• A-ESBL- all Pn + 1ST/2ND/3RD cephalosporin + monobactam
• Overcome by b lactam + b lactamase inhibitor
• B-MBL- AMPc + imipenam
• C- AMP C b lactamase- esbl + cefoxitin & cefotetan
• D- oxacillinase

SALMONELLAE
 Primary intestinal parasites of man & animals
 Isolated from blood & intestinal tracts of vertebrates
 Found in sewage,rivers& other waters & also assd. with foods

 Needs tryptophan for growth
MORPHOLOGY ≡ GNR—
Motile except---S.gallinarum,S.pullorum
Non capsulated ,Non sporing

ENRICHMENT MEDIA
Liquid media to assist isolation of Salmonellae from
FAECES, SEWAGE, FOOD STUFFS
Inoculation6-18hrss/c Selective mediapure Salm.
1)TETRATHIONATE BROTH
Enriches Salmonella spS.typhi “& Shigella
Permits growth of Proteus also
2)SELENITE F BROTH
Most useful for Salmonella & Shigella spp.
3. Gn broth
DIFFERENTIAL &SELECTIVE MEDIA
1)MacConkey Agar & DeoxyCholate Agar
Non lactose Fermenting
Pale colonies Black center in 48hrs
2) XLD – PC with BC
3) Wilson Blair Media - BC

Biochemical reaction
• H2S +ve in TSI except strains of S. paratyphi A,

• citrate + except para A & S. Cholerasuis
• Salmonella do not ferment lactose (NLF)
• Antigenic str. 2399 serotypes (kaufmann white)

• O (Somatic O)- heat sable, less Agnic, low
Ab & slow
• H (Flagellar H) – heat labile & highly Agnic
Heat labile alcohol labile. Exist in two alternative
phases H1 &H2
Vi Ag – surface polysaccharide covering O Ag
• Heat labile when present it renders the bacterium

inagglutinable by O antiserum
• Possessed by S Typhi, dublin & S. Paratyphi C , Citrobacter

• Poorly immunogenic, but protective
• Absence of Vi Ab- poor prognosis
• Persistance in coalescent stage – carier state
• Epidemiological typing of S.Typhi- by Vi specific
bacteriophage
Lab diagnosis
• Enteric fever: step ladder-fever, rose spots, coated tongue, relative brady
cardia, HSM, epistaxis
• Pathogenecity – Attaches to payers pathches

• C/C 3-4 wks, GI hge
• Culture – blood, faeces, urine, bone marrow, rose spots etc
• Blood culture
• 90% +ve 1st week
• 75% +ve 2nd week
• 60% +ve 3rd week
• 25% +ve after till the fever develops
• Stool during first week organisms isolated in half the cases

• 3rd to 5th week – most easily isolated
• Urine  positive generally in the second & 3rd week in 25% cases
• Demonstration of circulating Ag – Coagglutination ,ELISA,CIEP
Detection of Antibodies (Widal)
• Antibodies appear by 7th to 10th day of illness
•  till the 3rd or 4th week after which declines
• Demonstration of 4 fold rise highly significant between 1st &

3rd week
• 1: 100 of O agglutinins  significant
• 1: 200 of H agglutinins  significant
• H agglutinins persist after vaccinations but more sp.

• O agglutinins disappear i.e within 6 weeks – s/o acute infec
• Pts treated with chloramphenicol show poor agglutinin

response
New Diagnostic tests
• IDL Tubex
• Typhidot & Typhidot M
• IgM dipstick test.

• Carriers  Incidence of chronic carrier is lower in
paratyhoid than typhoid fever, carrier, harbor the
organism in biliary tract, gall bladder & rarely in
intestine & urinary tract
• Widal no value in detection of carrier in endemic

countries like India
• Vi agglutinins (1:10) or more indicate carrier state

• Vi +ve in – paratyphi, dublin, citrobacter also
• TT: Ciproflox
• MDR – cipro, CTX, Azithro
• Carriers : Ax/ cipro
• NARST: - CTX, Azithro
• India no Dr to CTX
• R usually plasmid mediated
• Salmonella septicemia – typically caused by S.
cholerasuis
Salmonella gastroenteritis
• Zoonotic food poisoning

• Food like meat, egg , milk
• S. typhimurium, commonest species (30-40%)

• Others- S. enteritidis, S dublin, S. Newport,
S.Heidelberg
• Stool culture
Typhoid Vaccines
• TAB Vaccine
• Ty2 1a oral vaccine
• Typhim-Vi injectable vaccine
Future vaccines
• Vi-rEPA
• CVD-909
• S.typhi Ty2 with mutation deletions in different genes
TAB Vaccine
• Was used earlier
• Consisted of S.Typhi 1000 million and S. Paratyphi A & B 750

million per ml, killed by heating at 50-600C & preserved in
formalin.
• Vaccine is given in 2 doses of 0.5 ml subcutaneously 4-6 weeks

apart
Ty2 1a (typhoral) vaccine
• Live attenuated vaccine, available as enteric coated capsule or
liquid formulation.
• It consists of a mutant virus which initiates infection but self

destructs after 4-5 cell divisions.
• Three doses 2 days apart are taken on an empty stomach.
• Elicits protection from 10-14 days of the third dose.
• 65-96% protect for 3-5 yrs

Typhim Vi
• Injectable vaccine containing purified Vi polysaccharide.
• Given as a single subcutaneous or i.m. dose.
• Protection begins 7 days after infection.
• Approved for use in children more than 2 years of age.

SHIGELLA
• NM, NLF
• Bacillary dysentry
• MEDIA  DCA, XLD, SS, HE Agar
• E’MENT – Sel F, Sh. Dysentriae, G.N borth

• Sd does not ferment manitol, while rest 3 ferment
•
S dysentrieae
13 serotypes ;
• 1 – Shiga bacillus - Cat negative
• 2. Schmitzi-,
• 3-7 large sachs group

Sh. flexneri – 6+2 variants – MC IN India
Type 6 88
New castle biotypes
Manchester
Sh. boydii - 18 Serotypes

Less commonly associated with dysentry
Sh. sonnei  LLF – antigenically homogeneous

2 phases,
26 colicin types  (in US)
TOXINS OF SHIGELLA
ENDOTOXIN
• Polysaccharide lipoprotein complex of cell wall are

responsible .
• Produced by all Shigella spp.

toxins contd.
EXOTOXIN
• Also k/a Shiga toxin.
• Example of an exotoxin produced by a Gram Negative
Bacillus.
• Produced by S dysenteriae type 1.
• Acts as neurotoxin, enterotoxin & cytotoxin
• Neurotoxin : though k/a neurotoxin ,the primary site of

action is not the nervous tissue but the blood vessels ,
mainly of CNS with the neurological effects being
secondary.
• Enterotoxin: These are heat labile proteins act on
intestinal mucosa & cause transudation of fluids.
exotoxin contd.
CYTOTOXINS
• Responsible for cytopathic changes in cultured vero cells.
• Same as verotoxin 1 (shiga like toxin) produced by certain

strains of E coli(VTEC).
• Toxin has active A subunits (A1 & A2) & binding B subunits.
• Fragments A1 inactivates host cell 60 S ribosome.

Dysentery – Incubation Period – 2-3d
Dose 10-100 bacilli
Complications:
• Arthritis
• Toxic Neuritis
• Conjunctivitis, Parotitis
• Intususception
• HUS  associated with complement activation & DIC
• Bacillary dysentery - E coli, shigella, vibrio

parahemolyticus, C. jejuni
Shigella can be differentiated from E.Coli by all
of the following features except- (AI 99)
a. Shigella does not produce gas from glucose
b.Shigella does not ferment lactose
c. Shigella does not ferment mannitol
d.Shigella has no flagella is non motile
266
• Causes of HUS are ?
a) Shigella
b) EHEC
c) Campylobacter
d) Vibrio
e) Chemotherapy ( mito,cisplatin,bleo)
f) Ca breast
Vi antigen found in - (PGI June 05)
a. Salmonella paratyphi ‘A’
b.Salmonella paratyphi ‘C’
c. Salmonella Dublin
d.Klebsiella pneumoniae
e.Citrobacter fragilis

• An infant is brought to the ER with hemolytic
uremic syndrome and thrombocytopenia.
Stool sample was cultured and pale colonies
grown on Sorbitol MacConkey agar, which also
fermented glucose with gas. Which one of the
following bacteria would most likely be
isolated from a stool specimen?
– Shigella
– Salmonella
– Aeromonas
– E. coli 0157/H7
All are correct regarding widal test, except-
(AIIMS Nov 09)
a. Baseline titres differ depending on the
endemicity of the disease
b. High titre value is a single widal test is not
confirmative
c. O antibody last longer and hence is not indicative
of recent infection
d. H antibody cannot differentiate between types

• Which of the following is not true about
HUS? (AI 2012)
– May present with hemorrhagic colitis
– Serotoxin has no role in HUS
– Usually self-limited
– Fever is typically absent
• Which of the following is not true about
EHEC?
– It secretes serotoxin
– Affects children
– Causes HUS
– O157:H7 serotype is the less likely to cause HUS
Yersinia
• Belongs to family Enterobacteriaceae
• Yersinia pestis
• Y. enterocolitica
• Y. pseudotuberculosis
Morphology
• Gram negative coccobacilli
• Single or pairs
• Bipolar staining- safety pin appearance when stained
with geimsa and methylene blue.
• Non motile
• Non sporing
• Pleomorphism
• Y pestis- typical capsule (F1 gp antigen complex)
Bipolar "safety pin" staining of Yersinia
pestis (Giemsa stain)
Cultural characteristics
Nutrient Agar: Transparent pin point
colonies
Blood Agar : Small dark brown colonies
Broth :granular deposits at bottom
and sides of tube
: Stalactile growth with oil
poured on broth growth
Aerobic or facultative anaerobe optimum temp. 27C

Antigen structure
• Heat labile protein envelope antigen (F1)formed by
virulent strains.
• V and W antigens also formed by virulent strains
• Pesticin 1, fibrinolysin and coagulase.
• Endotoxin is similar to GNB

Human Disease
• Zoonotic disease- plague
• Continuum of illness
– Bubonic plague
– Septicemic plague
– Pneumonic plague
Bubonic Plague
• 80-90% of cases
• Incubation: 2-6 days
• Clinical signs
– Fever, malaise, chills, headache
– Bubo: swollen, painful lymph node
–  vomiting, abdominal pain, nausea
• Mortality (untreated): 50-60%
Septicemic Plague
• Systemic spread
• Clinical signs
– Similar to bubonic, plus
– Prostration, circulatory collapse,
septic shock, organ failure, hemorrhage, DIC
– Necrosis of extremities
• Microthrombi blocking capillaries
• Mortality (untreated): 100%

Pneumonic Plague
• Secondary - septicemic form spreads
• Clinical signs
– Fever, chills, headache, septicemia
– Respiratory distress
• Person-to-person possible
• Primary - Y. pestis inhaled
– Incubation: 1-6 days
– Without immediate treatment, it is fatal
Rat flea vector for plague
rat flea –wingless , hops < 2 ft

epicenters – Beed district (Maharashtra)
Sweat city (Gujarat)
Yersiniosis:
• Yesinia inf other than Y.pestis – i.e Y. pseudotuberculosis & Y.enterocolitica
• Both motile at 25c , not at 37c
• Cold enrichment done for culture
Y. pseudotuberculosis
• Zoonosis
• grows at 22c, urease+ve
• C/F - Mesenteric lymphadenitis, resembling appendicitis
Y.enterocolitica
• Causes acute enterocolitis and terminal ileitis, lymphadenitis.
• Reactive arthritis in HLA B27 pts.

• Which of the following about Salmonella Typhi
Vi antigen are correct except
– Total absence of VI antibody in a proven case of

typhoid carries good prognosis
– VI antigen provides a method of epidemiological
typing of S.Typhi by phage typing.
– VI antigen is strongly immunogenic.
- VI antibody is usually present in carriers
FRANCISELLA TULARENSIS
Tularemia: a disease of rodents, originally described Tulare
country, California
• Human infections:
– Tick bite
– Direct contact with rodents
– Inhalation of aerosols
– Ingestion of infected meat or water
• Capsulated, NM, GNR
• Found intracellularly in liver, spleen cells
• Fastidious
• Francis blood dextrose cystine agar
• Highly virulent strains: in N America,
• low virulence strains: Europe, Asia.
• Human disease:
– Local ulceration with lymphadenitis (most common
form)
– Typhoid like fever with glandular enlargement
– Influenza like respiratory disease
GENUS: LEGIONELLA
• Weakly Gram – negative
• Pleomorphic rods requiring cysteine and iron
• Water organisms L. pneumophila (and other legionellae)
Distinguishing Characteristics
• Stain poorly with standard Gram stain; gram – negative
• Fastidious requiring increased iron and cysteine for laboratory

culture (CYE, charcoal Yeast Extract)
• Diagnose: DFA (Direct Fluorescent Antibody) on biopsy, (+) Dieterle

silver stain sputum –ve for bact, pus cells ++
Antigen urine test for serogroup 1 only .Fourfold increase in antibody

Reservoir
• Rivers/streams/amoebae; air-conditioning water
cooling tanks
• Transmission - Aerosols from contaminated air –
conditioning , Aspiration
• No human – to human transmission
Predisposing Factors
• Smokers over 55 – years with high alcohol intake
• Immunosuppressed patients, e.g. renal transplant
patients
• Pathogenesis CMI- imp, HUM- not IMP
• Faculative intrcellular pathogen
• Endotoxin
Disease
• Seasonal
• Associated with air – conditioning systems,
Legionnaires’ Disease (“Atypical Pneumonia” – sput.- pus cell++ no

bacteria, dry cough) .
• Pneumonia ,Mental confusion, fever Diarrhea (no legionella in
gastrointestinal tract) in HAI cases mc sg -6
Pontiac Fever
• Pneumonitis ,No fatalities, SELF LIMITING mc sg -1
Treatment
• Fluoroquinolone or azithromycin or erythromycin with rifampin
for immunocompressed patients
• B lactum – no role
Prevention
• Routine decontamination of air – conditioner cooling tanks
EIKENIELLA CORRODENS
• lacks flagella & show Jerking or twitching motility
reqd haemin for aerobic growth
• Characteristic pitting or corroding of blood agar
• Pathogenicity – normal inhabitant of human mouth,

intestine & upper respiratory tract
• Occasionally causes OIs such as dental & periodontal
infection, sinusitis, otitis media, mastoiditis,
pneumonia, lung abscess, wound infection following
human bite, septic arthritis.
•  Bacteremia & endocarditis in

immunocompromised hosts, i/v drug users or
persons with previous valvular damage.
STREPTOBACILLUS MONILIFORMIS
• Gram negative, highly pleomorphic bacillus
• String of bead appearance, non motile, fastidious
• Pathogenicity  Normal inhabitant of the nasopharynx of rats,

causes streptobacillary rat bite fever also caused by Spirillum
minus (sodoku fever)
• When the organisms are acquired by ingestion of food, milk or

water contaminated by rat excrement it is known as Haverhill
fever
Lab diagnosis  Grow on culture media containing blood serum or
ascitic fluid
• L forms – fried egg appearance
• Mice are susceptible to Intraperitoneal inoculation.
• treatment – penicillin
• Oxidase positives
TAXONOMY:
• Controversial and rapidly evolving
• Family originally proposed by VERON in 1965
• According to the last edition of Bergey’s Manual For
Systematic Bacteriology :
Family VIBRIONACEAE included four genera
Vibrio
– Aeromonas
– Photobacterium
– Plesiomonas
MODIFICATIONS:
• Aeromonas Aeromonadaceae
• Plesiomonas Enterobacteriaceae
• Only two genera of Vibrio and Photobacterium

retained in the family
(Approved lists of Bacterial Names)
Non-Halophilic Vibrios
i) V. cholerae
ii) Non 01 V. cholerae
iii) V. mimicus
Halophilic Vibrios
i) V. parahaemolyticus
ii) V. alginolyticus
iii) V. vulnificus
Antigenic Classification of Vibrios
Vibrios
_________________________
Group A Group B
Cholera Vibrios Biochem & ag heterogeneous
Biochemically similar
Common H Antigen(Vibrio Cholerae)
________________
Serogroup O1 Non –O1 (O2-O139)
__________________________
Classical cholera Vibrios El Tor vibrios
______________ ____________
Ogawa Inaba Hikojima Ogawa Inaba Hikojima

MORPHOLOGY
• Gram negative
• Comma shaped
• Actively motile with polar flagella
• Darting motility
CULTURAL CHARACTERISTICS
• Strong aerobic
• Alkaline pH
HOLDING & TRANSPORT MEDIA
a) Venkatraman- Ramakrishnan media

Salt + Peptone + DW (pH=8.6-8.8)
b) Cary Blair medium
NaCl + sod. Thioglycolate+ CaCl2 (pH=8.4)

ENRICHMENT & TRANSPORT MEDIA
a) Alkaline Peptone Water (pH=8.6)
b) Monsur’s Taurocholate tellurite Peptone Water

(pH=9.2)
Selective MEDIUM
a) Bile Salt Agar : Water drop colonies (pH=8.2)
b) Monsur’s Gelatin Taurocholate tellurite

trypticase soy agar (pH=8.5)
c) TCBS : Yellow colonies (pH=8.6)

BIOCHEMICAL REACTION
CHO Fermentation: +ve with Gluc, Mal, Mannitol,

Mannose, Sucrose with acid only
Oxidase : positive
Indole : positive
Nitrate : positive
Catalase : positive
Nitrosoindole test : peptone water growth
+ H2SO4 (Pink colour)
String test
PATHOGENESIS
• Oral ingestion
• Adhesion to intestinal epithelial Cells
• Multiplication and production of Cholera toxin (CT)
• CT responsible for pathogenesis
• CT is similar to heat labile toxin of E. coli

• VC- Chromosomal (DNA coding), Ec- Plasmid (DNA coding)
• Toxin has A (A1+A2) +B subunit

Cont..
• Binds to ganglioside receptor of epithelial cells
• A2 which facilitates binding of the A1to B subunit
• A1 subunit activates adenyl cyclase
• Activation of adenyl cyclase cATP to cAMP
• Increased cAMP leads to hyper secretion of

electrolytes and water into the lumen of the intestine
• As a result "rice water" diarrhea of cholera

Binding of subunit B to Entry of A1 in cell and activation
GM1 ganglioside receptor Of adenyl cyclase
Accumulation of c AMP in the cell Secretion of Na+, cl-, K+, H2O,

HCO3 out of the cell
V.Cholerae O139 BENGAL
Appears to be a hybrid of the O1 strains and the

non O1 strains
CT indistinguishable from El Tor VC O1 strains
• Identified by:
(1) absence of agglutination with O group 1
specific antiserum
(2) agglutination with O group 139 specific
antiserum
(3) the presence of a polysaccharide capsule
• Because of these differences, pre-existing
immunity to V. cholerae O1 El Tor does not
protect against the Bengal infection
Diff. between classical & El Tor
Test Classical El Tor
Haemolysis - +
VP - +
Chick red cell - +
agglutination
Polymyxin B St. + -
Group IV Phage + -
susc.
El Tor phage 5 _ +
susceptibility
• string test positive vibrios?
– Vibrio chlolerae
– Vibrio parahaemolyticus
– Vibrio damsela
Treatment-
• DOC in adult- Doxycycline ( if resistant, then cipro)
• DOC in children- cotrimoxazole
• DOC in pregnancy- Furazolidone
• DOC for chemoprophylaxis- Tetracycline
• Vaccines - Killed whole organism & Oral Vaccine –
• 50-60% protection for 3-6m, adult > children
• No cross protection b/t ogawa & inaba
• Cross protection b/t ElTor & Classical
Vibrio parahemolyticus
• Food poisoning due to sea fish consumption

• Grows only in media with NaCl (8%)
• Gram neg comma shaped with bipolar staining
• TCBS : Green colonies with opaque centre
• B/c reactions similar to VC exc. Sucrose –ve
KANAGAWA PHENOMENON:
Strains showing hemolysis on high salt agar
(Wagatsuma Agar) are pathogenic
Nonpathogenic ones don’t show hemolysis
Pathogenesis: Enteritis by invasion of intestinal epith.

• V. parahemolyticus does not show any hemolysis on
sheep blood agar but is ß-hemolytic on Wagatsuma agar
which contains human blood.
• This phenomenon is named as KANAGAWA
PHENOMENON and is associated with virulence The
strains are considered pathogenic .
Swarming :
• Seen on solid/highly viscous media
• Vibrios produce elongated swarmer cells that produce

both polar and peritrichous flagella. The latter are non
sheathed and have a diff. wavelength from that of polar
flagella.
• Swarming shown by: V. parahemolyticus,, V. alginolyticus,

Vibrio alginolyticus
• Found in sea fish
• Resembles V.parahemolyticus
• Salt tolerance upto 10%
• Assoc. with marine wound infection of eyes, ears

Vibrio vulnificus
• Lactose fermenter
• Salt tolerance <8%
• Assoc. with wound infection &

cellulitis following exposure to
sea water
• After ingestion of raw sea food

penetrates gut mucosa enters
blood stream causes septicemia
V.vulnificus infection in a 75 yr old with cirrhosis and
development of septic shock and bacteremia
• Aeromonas  inf in cold blooded aquatic
animal like frog – red leg disease
• Causes of Gastroenteritis / food poisoning

Bact Food IP
Salmonella typhimurium Poultry eggs 18-36hrs
B. Cereus Toxin Fried rice 1-6 hrs
Clostridium perfringens Reheated meat 18-16 hr

toxin
C. botulinum toxin Canned food 1-2 day
(botulism)
S. aureus toxins Milk prod. 5-6 hrs
Campylobacter jejuni Poultry 2-11 days
) EH EC (VTEC) 0157 H7 026 Ham burg 1-5 days
Yersinia enterocolitica Raw pork 4-7 days

Vibrio parahaemolyticus Shell fish 1-2 days
Aeromonas Fresh water fish 2-4 days
True about enterotoxigenic e coli
• Not a common cause of travellers diarrhea
• Spread by fomites and hand to hand contact
• Common cause of watery diarrhea in children
in developing countries
• Causes invasive diarrhea
• One of the following bacteria does not show
swarming?
– Vibrio alginolyticus
– Clostridium tetani
– Proteus mirabilis
– Morganella morganii
• Which of the following statement is incorrect
– S typhi causes gastroenteritis
– S paratyphi produces acid and gas on glucose
– Vi antigen is related to virulence
– Persistence of Vi antibody indicates development
of carrier state
PSEUDOMONAS
• Gram – negative rod

• Oxidase – positive
• Aerobic nonfermenter
• Pseudomonas aeruginosa
• Pseudomonas Medical Ecology
• Pseudomonas aeruginosa is a ubiquitous water and soil.
Sources for infections include
• Raw vegetables, respiratory, humidifiers, sink drains, faucet
aerators.
• Transient colonization of colons of about 10% of people.

Bacteria get on skin form fecal organism.
• Requires exquisitely careful housekeeping and restricted diets

in burn units
CULTURAL CHARACTERESTICS
• GNB, motile, polar flagella, mucoid ext cellular
polysacch. alginate polymer, strict aerobe, 37ºC
• Nutrient agar : Large, smooth, translucent
colony, Pigment diffusion in
media
• MacConkey : NLF Colonies
& DCA
• Blood Agar : Large colony. may be haemoly
• Cetrimide Agar: Selective media
• Peptone Water: Surface pellicles
Pigment Production:
• Pyocyanin :Bluish green, phenazine

pigment, H2O & chloro. soluble
• Fluorescein:Greenish yellow, H2O soluble
• Pyorubrin :Reddish brown, H2O soluble
• Pyomelanin:Brownish
Biochemical Reaction
• Glucose utilized oxidatively

• Oxidase positive→30 sec.
• Catalase positive
• Arginine dehydrolase positive
• Gelatinase positive
Toxins and Enzymes:
i) Extracellular Products : Pyocyanin

Inhibits mitocho. enzy.
ii) Extracellular Enzymes : Haemolysin
protease, elastase,
phospholipase play a role in
formation of local lesion.
iii)Exotoxins: 2 A
S
Exotoxin A Polypeptide, Inhibits protein sythes
Endotoxin LPS. Pyrogenic reaction,

Schwartzman reaction.
• Most common cause of nosocomial pneumonia
• Most common cause of nosocomial wound infections

in burns
• Malignant ottitis externa
• Grape – like odor

• Slime layer
Reservoir- Ubiquitous in water
Pathogenesis:
• UTI (In dwelling catheter).

• Acute Purulent meningitis (LP).
• Pneumonia (ICU).
• Septicemia in burns, debilitated patients with
immunosuppressive drugs.
• Endocarditis (drug addict), septic arthritis.
• Otitis media, Eye infection, Ac neutralizing vasculitis.
• Infantile diarrhoea.
• Fever +shock+ skin lesions (black necrotic centre –
ecthyma gangrenosum)
Treatment:
• Intrinsically resistant to most antimicrobials
• Carbenecillin, Ticarcillin, Cefotaxime,

Ceftazidine,Gentamicin and Tobramycin,
Perflox, Oflox.
• Localized Infection: Topical colistin, Polymyxin

B or 1%acetic acid.
• A 50 year old chronic alcoholic male agricultural worker presented with
high grade fever of one week duration with spells of chills and rigor.
Examination of the respiratory system revealed bilateral crepitations with
scattered rhonchi. Multiple subcutaneous nodules were found on the
extensor surface of the left forearm, arm and left leg. Direct microscopy of
the pus aspirated from the skin nodule revealed plenty of Gram negative
bacilli with bipolar staining. Culture revealed distinct rough corrugated
grey-white colonies on Blood agar. The organisms were motile and oxidase
positive. The most likely diagnosis is -
(AIIMS Nov 03)
a.Plague b.Melioidosis
c.Bartonellosis d.Actinomycosis

Burkholderia mallei (P. mallei)
Glanders in animals (horses, mules)
Man acquires infection from animal.
Morphology : GNB, non-motile, beaded
faculta. anaerobe,
grow on ordinary media.
Animal Pathogenicity:
1. Glanders :Resp. system affected with
profuse nasal discharge.
2. Farcy :Infection through skin with
involvement of lymph vessels and lymph
nodes. “Farcy pipes”.
• Strauss reaction: intraperitoneal injection in male
guinea pig causes testicular swelling in 2-3days due
to bacillary invasion of tunica vaginalis.
• Humans:
• Skin abarasions or wounds which come in contact

with sick animal.
• Acute fulminant febrile illness
• Chronic indolent infection
BURKHOLDERIA PSEUDOMALLEI
• Causative agent of meliodosis (Glanders like disease)

• Gam negative rod, bipolar stained, motile, oxidase
positive
• Found in soil in south –east asia, north Australia
• Occurs in rats, guinea pigs, rabbits
• Human infection: Inhalation, inoclation, ingestion of
foodstuffs contaminated with excreta of infected
animals
• Agricultural workers mostly prone to infection
Humans
• Subclinical infection
• Pulmonary infections : similar to TB
• Multiple abscesses in various organs & tissues
• Fulminating septicaemia
• Also known as “ Vietnam Time-Bomb”
•
• India
• Reported form Maharashtra, Tamil Nadu, Orrisa, west Bengal,
Tripura.
• Laboratory diagnosis
• Culture
• Serology: ELISA, IHA
9. Toxin which interfere with protein synthesis is
– E coli toxin
– Pseudomonas toxin
– Staphylococcal toxin
– Cholera toxin
– Shigella
– C. diph.
All of the following statement about plague are
wrong except - (AIIMS 04)
a.Domestic rat is main reservoir
b.Bubonic is the most common variety
c. The causative bacillus can survive upto 10 years in
the soil of rodent burrows
d.Incubation period for pneumonic plague is one to
two weeks

• All the following statement of typhoid carrier
state are true except
– Carrier presents even after the treatment of
disease
– In carriers the organism rest in Gallbladder
– In Japan, urinary carrier state is common
– Detection of carrier state is by antibodies to “O” &
“H” antigens
• HAEMOPHILUS
– Blood loving
– Requirement for one or more of the accessory factors: X & V in blood
– “Pfeiffer’s bacillus” or H influenza
– Plemorphism
– Acute infection: capsulated strains
– Stain: Methylene blue or dilute carbol fuchsin
– X: hemin or porphyrin required : heat stable
– V: heat labile, inside RBCs NAD or NADP- ng on BA
– Staellitism
– Levinthal’s agar (iridescent colonies of capsulated strains),

Field’s agar (best for primary isolation)
Satellitism
– 8 biotypes
– Biotype 1 for menigitis (mc in less than 2 yr)
– Oxidase + Catalase +
– Refrigeration kills
Antigenic properties
• Capsule : a-f capsular types; 95% type b; (PRP/ribose or
ribitol) Hib PRP used in vaccine
• OMP
• LOS
Pathogenicity
Humans only
Invasive: meningitis, arthritis, epiglottitis, pneumonia, bacteremia,
endocarditis, Pericarditis: capsulated; children
Non invasive: secondary: otitis media, sinusitis, COPD adults; non-

capsulated
Meningitis: nasopharynx-blood stream; 2mths to 3 years
Epiglottitis: most common cause; blood culture
Pneumonia: infants; older children & adults – lobar
Hib PRP vaccine: not effective in <2 years of age
Rifampicin prophylaxis
• H aegyptius (Koch’s –Week’s bacillus)
– “Pink eye” conjuctivitis
– Brazilian purpuric fever
• H ducreyi
– Chancroid: painful, LNs
– “School of fish” or “rail road track” appearance
– Soft sore- tender and non indurated
– Medium used: chocolate agar with 1% isovitalex, Vancomycin
93mg/ml
• HACEK group of organisms

– Fastidious slow growing bacteria, normal or oral cavity
– Hemophilus species. Actinobacillus actinomycetecomitans.
Cardiobacterium hominis, Eikenella corrodens, Kingella
kingae.
Growth characteristics of Haemophilus
SPECIES GROWTH REQUIREMENT

CO2 X V
H. influenzae - + +
H. aegypticus - + +
H. ducreyi V + -
• All the following statements are true about
vibrio cholera except:
A .Non-halophilic
B .isolation can be done on ordinary media
C .Can suevive outside the human intestine
D .Man is the only reservoir of infection
• The Endotoxin do not play a role in
pathogenesis of?
– E.Coli
– V. Cholerae
– Klebsiella pneumonae
– Pseudomonas
• All of the following are true vibrio cholerae 0-139
except:
A Causes a disease indistinguishable from V. cholera

B Was 1st isolated from Chennai
C Has O polysaccharide capsule
D Antibodies against vibrio choleral 01are protective.
E. epidemiologically indistinguishable from 01 el tor
F. Expresses 01 Ag
Not true about H. influenzae
• Needs factor x and v
• Capsular polypeptide is the virulence factor
• Uncommon cause in less than 2 months
• Mc cause of invasive meningitis
• Vibrio el tor all are true except? (AI2010)
a. Humans are only reservior

b. Toxin acts other than intestine also
c. Killed by boiling for 3 min
d. Survives on ice for month
• Which of the following is associated with ear
infections ?(AI 2012)
– Vibrio alginolyticus
– Vibrio parahemolyticus
– Vibrio vulnificus
– Vibrio mimicus
Bordetella pertusis
Morphology
• Gram-negative
• Metachromatic granules on toludine blue stain
• small, ovoid, coccobacilli
• Non motile
• Non sporing
• Capsulated
• Arranged in clumps
• Thumb print appearance – culture film
Cultural characteristics
• Strict aerobes
• Growth best 35 -360 C
• Bordet gengou glycerin potato blood agar

• Charcoal blood agar
• Colonies on B.g.g.p. blood agar – bisected pearls or

mercury drops
• Confluent growth- aluminium paint appearance
other media  Regan Lowe, Lacey DFP medium

• All of the following are true regarding pertusis
toxin except (AI 2009)
•
– Its an exotoxin
– Shows phenomenon of piracy of adhesions
– Accumulates cyclic GMP
– Activation of insulin secretion
- sensitization of the body to histamine
- Leads to proliferation of lymphocytes
Antigenic structure and virulence factors
• Agglutinogens
• Pertusis toxin- exotoxin
• Filamentous haemagglutinin (FHA) – promotes sec.
infec. By H. inf.
• Adenylate cyclase
• Heat labile toxin
• Tracheal cytotoxin
• Lippopolysaccharides
Epidemiology
• Incidence highest in first yr of life

• Maternal antibodies don’t give protection
• Transmission by droplets and contaminated fomites
• Chronic carriers not known
• 95% cases - Bordetella pertusis
• 5% cases - Bordetella parapertusis
• Pathogenesis  Source of infection (whooping cough)
patient in early stage
• No healthy carrier
• Attack rate of more than 90% in non immunized individuals
• Specimen – per nasal swab for nasopharynx ( catt phase)
• DOC  EM for whooping cough
• B. pertusis, B. parapertusis – whoopingb cough
• B. bronchoseptica-(motile) – usually infects dog, man infected

rarely.
Pathogenesis
• Incubation period 1 to 2 weeks
• Three stages : catarrhal, paroxysmal,
convalescent
• Catarrhal : fever, cough, sneezing
(2 wks) maximum infective stage
• Paroxysmal : characteristic inspiratory gasp
(whooping cough 2-4 weeks), vomitting
and max c/c in this phase
• Convalescent : increased severity and
frequency of coughing(2-4
wks)
• Person having open wound is in leg comes in
contact with sea water & develop wound
injection organism implicated is
– V. Parahaemolyticus
– V. vulfinicus
– CI. Botulinum
– S. paratyphi
Brucella –
• Zoonosis Brucellosis – Malta & , Undulant fever
• GM NEG ; Coccobacilli  Goat, Sheep, Cattle
• Six Spp. B. Melitensis – Sheep, Goat – most

pathogenic
• B. Abortus – Cattle (Capnophillic)
• B. Suis – Pig
• B. Canis – Dog.
• Strict Aerobe
• Bipolar staing
Transmission to Humans
• Direct Contact
• conjunctiva or broken skin in contact with infected
tissues
• Blood, urine, vaginal discharges, aborted fetuses,
placentas
• Ingestion
• Raw milk & unpasteurized dairy products,rarely
undercooked meat
• Inhalation of infectious aerosols
o Pens, stables, slaughter houses
o Laboratory transmission
• No evidence of person-to-person transmission
Human Disease
• Undulant fever characteristic
• Intermittent or irregular fever with variable duration
• Nonspecific and variable symptoms
• Headache, weakness, arthralgia, depression,
weight loss, fatigue, liver dysfunction
• Any organ or system can be affected
• CNS or heart disease more difficult
• Media – Can grow on NA (slow)
• SDA
• BLD culture  Casteneda
• Culture- serum dextrose agar, trypticase soy

agar
• Ds of RES
• Serol – STD Aggl Test (SAT)- for IgM,

• Elisa CFT - chronic
• Brucellin Skin Test
• Animals   Rose Bengal Card test

• Milk Ring Test
• 
• Br. abortus stained with Haematox.
• +
• Milk (Suspected. to contain Ab)
•  700 C X 40 - 50 min
• Positive  Bact. Aggl. With milk
• 
• Rise with cream
• 
• Blue Ring
• False about non typhiodal salmonella
a. Causes serious systemic infection
b.Blood culture is more sensitive than stool for
diag of carriers
c. Poultry products are source of infection
d.Quinolones are efferctive
• A 12 yr girl presents to em with hypotention ,
inguinal& axillary lymphadenopathy &
stalactile growth on culture. Diagnoasis is ?
(AiIms2008)
• CAMPYLOBACTER JEJUNI
– Spiral, GN; single polar flagellum
– Thermophilic 420C, microaerophillic
– Gull wing appearance- s shaped
– 4-35% of fecal specimens of pts with acute diarrheal
disease, faeco-oral (raw milk, partially cooked poultry,
contaminated water); present in GIT of wid and
domestic animals.
Mechanism of diarrhoea
• Heat labile enterotoxins resembling CT
• Invasion like Shigella
• Cytotoxin
• Media
– Transport: Cary Blair medium
– Selective: Skirrow’s, Campy BAP, burazzlers
Campylobacter jejuni: A causative agent of Guillain
Barre Syndrome
Lymphocytes in Spinal Roots
• A demyelinating disorder resulting in acute neuromuscular

paralysis. Also known as Acute Post Infective Polyneuritis.
• An estimated one case of GBS occurs for every 1,000 cases of
campylobacteriosis and up to 40% of patients with the
syndrome have evidence of recent Campylobacter infection.
The following statements are true regarding
melioidosis except - (AI 05)
a.It is caused by Burkholderia mallei
b.The agent is a gram negative aerobic bacteria
c. Bipolar staining of the aetiological agent is seen
with methylene blue stain
d.The most common form of melioidosis is
pulmonary infection

Helicobacter pylori
HELICOBACTER PYLORI
 GN spiral rod, motile unipolar tuft of lophotrichous flagella
 Microaerophilic, abundant urease production

 Mild acute gastritis, peptic ulcer disease, chronic gastritis
 Risk factor for gastric malignancy ( gast.antrum colonised)
Invasive Test
• Warthin starrey silver staining (histo path)
• Culture Skirrow’s medium (in sensitive)
• Biopsy urease test
Noninvasive -Urea breath test, Serology (ag & ab),stool ag
Virulence: Urease production, Cag (cytotoxin associated gene- pep

ulcer+G cancer ), vac (vacoulating cytotoxin gene- pep ulc)
Helicobacter pylori
• Pathogenesis :
Inhibits D cells secreting somatostatin- so decrease inhibition of
gastrin- hypersecreation of gastrin- metaplasia.- infla- ulcer
• 80% of deodenal ulcer and 60% of gastric ulcers are d/t H pylori.
• Increased risk of GA. Adenoca, and Ga MALTOMA

• Developing count- 80% develpoed – 30%
• TT
• 1- OCA x 7 days
• 2. OCM x 7 days
• 3. OBTM x 14 days
• PASTEURELLA MULTOCIDA
– GN, NM, oxidase+, Indole+, Mac: NG
– Sometimes commensal in human upper respiratory tract
– Human infections: animal bites or trauma
– Local suppuration following bite (wound infection, cellulitis,

abscess, Osteomyelitis), meningitis following
– head injury, respiratory tract infection, appendicitis.

• ANAEROBES
CLOSTRIDIUM
• Gram Positive, spore forming, Anaerobes.
Pathogens normally found in gut
• C. tetani
• C. Welchi
• Motility – stately
• All motile except- cl perfringens, cl. Tetni vi
• Cl perf. And butyricum - capsulated
• Only proteolytic- tetani
• Only saccharolytic – botu
• Neither- cochlearum
• Both – sporogens
• Predominant saccharolytic – welchi, novyi, septicum

Cl. perfringes (Welchi)
• Non Motile, Spores oval, subterminal
• Inhabitant of gut, found in feaces
• 4 Major (Lethal) Toxins - ,, ε, i  5 types (A-E)
• on BA  Target Haemolysis i.e. due to

• θ toxin → complete haemolysis
•  toxin → incomplete haemolysis
• Mc- by sertype A, resposible for toxemia of GG.
• Litmus Milk reaction (+)

• Naegler reaction (+) d/t  toxin  opalescence on egg
yolk media (inhibited) by antisera
• Also by Cl.novyi/sordelli/ bifermentans

Infections:
• (1) Gas gangrene – heatb labile toxin,
• IP- 4-6 Days
• 80% cases welchi, rest novi, septicum, histolyticum,
• Septicum- citron bodies (boat shaped wid irreg stain)
• (2) Food Poison – Type A  Enterotoxin
• (Similar to cholera & EPEC) – heat stable spores
• 3. NEC - Necrotising enteritis (Pigbel): type C
Cl. Tetanus
• Motile, SWARMING – drum stick appearance
• (type VI – non motile)
•
• Drum Stick Appearance (Terminal Spores)
• Two toxins : - Fur tree appearance
• (1) Tetanospasmin – Neurotoxin (exotoxin) - inhibit
Normal inhib. reflex --( -bits release GABA) Spasm ( presynaptic)
• Trismus – 1st symp., Inc. msl tone rigidity, Mentation unimpaired,
deep tendon reflex↑
• 2) Telanoysin
Clostridium botulinum
 8 types (A-G), A,B,. & E mc cause of human ds
 A-E – neurotoxin, C2 – cytotoxin.
 Type C,D- bacteriophage coded
 Blocks ACH- acts on PNS, desending palsy
• Botulinum toxin  Most toxic Known to man kind
• Min Lethal dose  1 g ( release after cell death)
• Neurotoxin  inhibit release of Acetylcholine
• Toxin used for t/t of strabismus, blepharospasm
• DDD , ptosis, consti, DTR – Normal, descending paralysis
(1) Food borne bolulism  Preformed toxin (h.labile)

Canned food  bulging after 18-36 hours
(2) Wound botulism
(3) Infant botulism (spore ingesn- h.stable ) - Floppy Child s.
Cl. Diffcile
 Psudomenb. Colitis, due to antibiotics like
S/S diarrhoea, no RBC in stool
Toxin neutralized by Cl.sordelli anti toxin
Clindamycin, Ampicilin
• Rx – Vancomycin , MNZ
• Toxins A  Enterotoxin – diarrhea

• B  cytotoxin
• Diag- stool culture
• Elisa
• A 25 yrs old male on chronic antibiotic therapy
presented with fever, abdominal pain &
diarrhoea associated with occasional blood in
stools. Which among the following are true?
– Easily isolated in culture
– Presence of large central spores ( Ter)
– Toxin is neutralized by Clostridium sordelii
antitoxin
– Vancomycin is the drug of choice - M
Regarding gas gangrene one of the following is
correct - (AI 04)
a. It is due to clostridium botulinum infection
b. Clostridial species are gram-negative spore
forming anerobes
c. The clinical features are due to the release of
protein endotoxin
• Gas is invariably present in the muscle
compartments
Regarding Clostridium tetani, all are true
except?
(AI2011)
A. Spores are resistant to heat
B. 2 doses give immunity in primary immunization
C. Incubation period is 6-10 days
D. Person to person transmission does not occur

Clostridium perfringens except- (AI 05)
a. It is the commonest cause of gas gangrene
b. It is normally present in human faeces
c. The principal toxin of C. perfringens is the alpha
toxin
d. Gas gangrene producing strains of C.perfringens
produce heat resistant spores (sen)

Regarding Clostridium perfringens gas
gangrene, false is - (AIIMS Nov 10)
a. Common cause of gas gangrene
b.Nagler reaction positive
c. Most common toxin is Hyaluronidase
d.Food poisoning strain of Cl. Perfringens
produces heat resistant spores

• Pseudomembranous colitis, all are true except-
(AIIMS May 07)
a. Toxin A is responsible for clinical manifestation
b. Toxin B is responsible for clinical manifestation
c. Blood in stools is a common feature
d. Summit lesions is early histopathological finding

• True regarding pseudo membranous colitis are
all except - (AI 2000)
a. It is caused by clostridium difficile
b. The organism is a normal commensal of gut
c. It is due to production of phospholipase A
d. It is treated by vancomycin

Site of action of tetanus toxin - (AI07)
a. Presynaptic terminal of spinal cord

b.Post synaptic terminal of spinal cord
c. Neuromuscular junction
d.Muscle fibres

A person has received complete immunization against
tetanus 10 years ago. Now he presents with a clean
wound without any lacerations from an injury
sustained 2.5 hours ago. He should now be given-
(AI 01)
a. Full course of tetanus toxoid
b. Single dose of tetanus toxoid
c. Human tet globulin
d. Human tet globulin and single d
dose of toxoid

• The bowel contains many microorganisms but
the most prevalent bacterium is
– B. fragilis
– Lactobacillus
– S. epidermidis
– E. coli
Bacteroides fragilis
• Anaerobic, gram –negative rods

• Anaerobes are identified by biochemical test and
chromatography
• Reservoir - Human colon;(10 11/g) the genus Bacteroides
in the predominant anaerobe.
Pathogenesis
• Modified LPS (Missing heptose and 2-keto-3
deoxyocotnate) has reduced endotoxin activity
• Capsule is antiophagocytic
Septicemia, peritonitis (often mixed infections), and abdominal
abscess
• Treatment
• Metronidazole, Clindamycin or cefoxitin. Abscesses should be
surgically drained.
• Antibiotic resistance is common (penicillin G, some cephalosporins,

and aminoglycosides)
• Prevention
– Prophylactic antibiotics for gastrointestinal or biliary tract surgery
• Bacteriodes melaninogenicus = Prevotella melaninogenica

• Melanin-producing (black) Bacteriodes)
• Normal gingival flora
• Oral abscesses
• Heparinase leads to clotting in brain.
50, With reference to bacteroides fragilis the following
statements are true, except:
– B. fragilis is the most frequent anaerobe isolated from

clinical samples
– B. fragilis is not uniformaly sensitive to metronidazole
– The lipopolysaccharide formed by B. fragilis is
structurally and functionally different from the
conventional endotoxin ( lack hydroxymyrsticacid)
– Shock and disseminated intravascular coagulation are
common in bacteroides bacteremia
– ( less potet edotoxi so less DIC )
• A man with cystic fibrosis is admitted in the
hospital with pneumonia. Hs sputum is
mucoid and greenish in color. On gram staing
it was a Gram negative rod. Which of the
following organisms should be suspected.
– Klebsiella
– Streptococcus pneumonia
– Pneumocystis jirovecii
– Pseudomonas aeruginosa
•
• All are true about Bacillus anthracis except
– Capsule is made up glutamic acid
– Bamboo stick appearance in gram staining
– Hemolytic colony in blood agar
– Sudan black stain used to demonstrate lipid
granules.
• The entire following are true of Botulism.
Except;
– It causes a symmetric descending paralysis
– Botulinum toxin acts on GABA neurotransmitter.
– All produce neurotoxin except C2 which shows
enterotoxic activity
– Cl. botulinum can be isolated from food or the
patient’s feces.
SPIROCHETES
• Pathogenic Treponema
• Borrelia
• Leptospira
• nonhogenic spirochetes
Spirochaetales Associated Human
Diseases
Genus Species Disease
Treponema pallidum ssp. pallidum Syphilis
pallidum ssp. endemicum Bejel
pallidum ssp. pertenue Yaws
carateum Pinta
Borrelia burgdorferi Lyme disease (borreliosis)
recurrentis Epidemic relapsing fever
Many species Endemic relapsing fever
Leptospira interrogans Leptospirosis
(Weil’s Disease)
Epidemiology of Borrelia Infections
Pediculus humanus
Borrelia
recurrentis
Ornithodoros spp.
Borrelia spp.
Ixodes spp.
Borrelia
burgdorferi
• Borrelia
(1) Epidemic or louse borne relapsing fever caused by B

recurrentis
(2) endemic or tick borne relapsing fever – caused by B.

duttoni, B. hermesii
Relapses d/t Ag variation, d/t vMP gene.
• Microscopic examination  Blood – wet film under dark
ground or phase contrast microscope
• Thick & thin smears Giemsa & Leishman stain

• mice – intra peritoneal infection
Pathogenesis of Lyme Borreliosis
 Lyme disease characterized by three stages:
i. Initially a unique skin lesion (erythema chronicum migrans
(ECM)) with general malaise
 ECM not seen in all infected hosts
 ECM often described as bullseye rash
 Lesions periodically reoccur
ii. Subsequent stage seen in 5-15% of patients with neurological

or cardiac involvement
iii. Third stage involves migrating episodes of non-destructive,

but painful arthritis
Erythema chronicum migrans of Lyme
Borreliosis
Bulls’eye rash
• B. burgdorferi  First identified in 1975 in Lyme Connecticut USA
• Lab Diagnosis – Diagnosis mainly on clinical ground.

erythema chronicum migrans (ECM) at site of vector bite,
followed by expanding bright red rash suggestive of positive
diagnosis.
•  Isolation & identification of B. burgdorferi from skin lesions or

blood.
• Culture too slow, low yield culture on Noguchi medium
• Microscopic detection – Dark ground, phase contrasts,

Immunoflorescence ,silver staining
• DNA probes
• IgM and IgG Ab in serum, csf
Borrelia spp
Borrelia vincenti
• Normal mouth commensal.
• Can cause ulcerative gingivostomatitis or

oropharyngitis (vincent’s angina) in
immunocompromised or malnurished.
• May be seen in lung abscess, skin ulcers & gangrenous

balanitis.
• Vincents angina – T vincentii is often associated with
fusiform bacillus known as Leptotrichia buccalis.
• Which is also known as Fusobacterium fusiformis

• T. Pallidum sub spp. Pallidum – Syphilis
Nonveneral
1. T. pertenue : Yaws
2. T. endemicum : Endemic Syphilis
(Bejel)
3. T. carateum : Pinta
Can’t d/t all these morphologically – PCR

• T. pallidum  Motile (fine fibrils)

Lashing Motility Flexn / Extension
Corck screw
• Dark Ground Mscope
• Silver Impreg. :Levaditi (tissue), Fontanas (smear)

Immuno Fluo (DFA)
• Can’t Be cultivated on artificial media

• Nicole strain : virulent – testicular pasage in rabbit
• Reiter strain – non vir.
CLINICAL DISEASE – 4 stages
• Primary syphilis
• Secondary syphilis
• Latent syphilis
– early and late
• Tertiary syphilis
– occurs after several years
Primary Syphilis
• Incubation period :10-90 days
• Invasion of mucus membranes, rapid multiplication & wide

dissemination
– Occurs prior to development of the primary lesion
• inflammatory response at the site of inoculation (chancre

(usually painless)
– profuse shedding of spirochetes
– Primary lesion heals spontaneously by fibrotic walling-off

within two months, leading to false sense of relief
Secondary Syphilis
– Secondary disease (6-8 wks) weeks after primary
lesion
– Widely disseminated mucocutaneous rash
– highly contagious
– Condyloma lata
Generalized Mucocutaneous Rash of
Secondary Syphilis
Latent Syphilis
• Refers to period following disappearance of symptoms of
secondary syphilis
• Patient is asymptomatic but has the infection and serologic

tests are reactive
• Early latent syphilis: <1 year

• Late latent syphilis: >1 year
• Diagnosis is based on a combination of serologic results and

past history of symptoms of primary and secondary syphilis
– About 40% of late latent patients progress to late tertiary syphilitic
disease
Tertiary Syphilis
• Syphilis will progress to tertiary stage in 30% of untreated
patients
• Tertiary syphilis characterized by localized granulomatous

dermal lesions (gummas) in which few organisms are
present
• Non-treponemal tests may be negative in 30% of cases
• Treponemal tests are almost always positive and may be the

only basis of diagnosis
Tertiary Syphilis
• Late tertiary -: Neurological manifestations such

as Tabes dorsalis and General paralasis of the
insane seen decades after the initial infection
• Cardiovascular syphilis: -
– Symptoms of aortic insufficiency or aneurysm
– Reactive treponemal tests
– No H/O treatment for syphilis
Neurosyphilis
• T. pallidum invades the CNS early during the course of
infection and may lead to meningeal inflammation
• Syphilitic meningitis and meningovascular syphilis seen in

secondary syphilis
• Parenchymatous neurosyphilis occur in tertiary syphilis
• Diagnosis of neurosyphilis requires

– Reactive treponemal test with serum sample
– CSF leucocyte cell count of >5 mononuclear cells/cmm
– Reactive VDRL-CSF test
– Demonstration of T. pallidum in tissue stained by silver stain or
fluorescent antibody or PCR
Congenital syphilis
• Transplacental transmission can occur at any stage of
pregnancy
• Lesions of congenital syphilis develop only after 4th month of

gestation
• Congenital syphilis can be prevented by

– Screening pregnant women for syphilis (in first trimester)
– Giving adequate treatment before 4th mth of
pregnancy
• Early -Rhinitis, snuffuls, osteochondritis, Bulla
HSM, jaunice
• LATE: clutton joint , 8th n palsy,
• Residual : hutchinsons teeth, mulburry molar,

saddle nose, saber tebia, rhagaddes
• Lab Diagnosis : In Exudates : Darkgrd microscope, DFA
• Serology : (A) Non Trep Test : (1) Wasserman (CFT)

(2) Kahn Test – Tube Flocculation
(3) VDRL – Slide Flocculation
(4) RPR
AG  Cardiolipin (Diphosphatidyl Glycerol) + Lecithin + cholester.
(B) Specific Trepon : TPI (Immob.)-[live] (using nichole st)

(Killed)  TPA (agg).
TPIA (Immune adherence
FTA, FTA-ABS ( 1st to be +ve)(7-10 d)
• (C) Specific Using Reiter strain RP CFT.

Darkfield Microscopy of Treponema
pallidum
Biological false positive reactions
Acute Chronic
Last few weeks or Last longer than 6
months months
Acute SLE, Collagen
infections,injuries, vascular diseases,
inflammation and early leprosy ,malaria
HIV infection relapsing fever,
hepatitis, Infectious
mononucleosis,
tropical eosinophilia
Yaws
• In India cases seen in Andhra Pradesh, Orissa & M.P.
• Primary lesion: extra-genital papule, usually on legs,

enlarges and ulcerates
• Late- HSM,destructive gummatous lesions of bones
• Infection transmitted through direct contact of traumatised

skin with exudate from early yaws lesion of patient,
probably by flies also
Pinta
• Non-venereally transmitted infection due to T. carateum
• Manifestations confined to skin only
• Skin lesions are non-destructive (pruritic lesions )

• Spread by direct contact with infectious lesions
GENUS LEPTOSPIRA
Species
L. Interrogans
L. biflexa
L. parva
• L. interrogans – parasitic strains
• L. biflexa & L parva } free living saprophytic strains

Silver Stain of Leptospira interrogans serotype
icterohaemorrhagiae
 Obligate aerobes
 Characteristic hooked ends (like a
question mark, thus the species epithet
– interrogans)
PATHOGENESIS
L interrogans causes Leptospirosis.
• 3R – rat urine, rain fall, rice fields

Clinical picture in Man
• Incubation period – 10 days
• Mild pyrexia to fatal illness with hepatorenal damage ( Weil’s

disease)
• L interrogans serovar icterohaemorrhagiae causes most

serious illness (Weil’s disease).
• Fever with rigors, vomiting, headache, jaundice in 10 – 20%

cases by 2 to 3 days.
• Purpuric hemorrhages on skin and mucus membranes.

• Albominuria is a constant feature.
3) Leptospira
• Leptospira may be demonstrated in blood & in urine by Dark

ground microscopy, phase contrast, silver impregnation
• Blood examination useful in early phase as disappear from

blood after 8 days , in the urine in the 2d wk & ntermittently
thereafter for 4 – 6 weeks.
• Culture media Korthof’s medium , EMJH, Fletchers
• Serological Diagnosis
• Genus specific – CFT, ELISA
• Sero group & serovar specific  macroscopic & microscopic
agglutination tests.
Comparison of Diagnostic Tests for
Leptospirosis
TREATMENT
• Penicillin is the most effective treatment for

leptospirosis.
• In cases of allergy to penicillin streptomycin,

tetracyclin or erythromycin can be used.
• A man presents with a maculopapular rash. He gives a history
of previous painless rash. Infection is due to? (AI2011)
A. Treponema pallidum
B. Chlamydia
C. Calymmatobacterium granulomatis
D. Haemophillus ducreyi

• About yaws all are true except: (AI2011)
A. Caused by Treponema pertenue
B. Transmitted non-venerally
C. Secondary yaws can involve bones
D. Last stages involve heart and nerves

• Which of the following is not used to diagnose Leptospirosis?
(AIIMS may 2010)
a. Microscopic agglutination test
b. Dark field microscopy
c. Macroscopic agglutination test
d. Weil-felix reaction

• All are causes of endemic relapsing fever
except
A. B. duttoni,
B. B. hermesii
C. B. Recurrentis
D. B. buccalis
• False about hemophillus influenzae is?(aiims
2009)
a. Requires X and V factor

b. Capsule plays an important role in
pathogenicity
c. Rare cause of meningitis in first 2 mths of life
d. Most common cause of meningitis.
21. Virulence factors of Bordetella pertusis are
– Tracheal cytotoxin
– Adenylate cyclase
– Pertusis toxin
– Filamentous hemagglutinin
– All of the above
Mycoplasma & Ureaplasma
• Very small measuring 0.2 -0.3 m in dia.
• can pass through bacterial filters
• Flagella ,pili –absent but some strains show gliding mobility due
to special tip like structure.
• Lack rigid cell wall, single trilaminar cell membrane.
• Mycoplasma & ureaplasma cannot synthesize their own
cholestrol & require it as a growth factor in culture medium
• Pleomorphic
• Stain with Dienes or Giemsa stain

• Stain poorly with Gram stain and are gram negative
• Fried egg colonies
• Culture medium ----- PPLO broth (bovine heart infusion broth
+ 20% horse serum + 10% fresh yeast extract)
• 200-500 m – large colony Mycoplasma

• 15-60m- small colony Ureaplasma
• Mycoplasma pneumoniae  commonly cause

tracheobronchitis pharyngitis, sinusitis, primary atypical
pneumonia (PAP).
• M pneumoniae may also cause extrapulmonary lesions

• Arthritis, meningoencephalitis, Transverse myelitis,
• Gullein Barrre Syndrome – haemolytic anemia
• Myocarditis – pericarditis
• Ureaplasma urealyticum - cause NGU, non chlamydial
urethritis, epididymitis, Vaginitis & cervicitis
• C. trachomatis responsible for 30-50% NGU
• cause chorioamnionitis, prematurity, post partum
endometritis
• Chronic lung disease of premature infant
• Ureaplasma isolated from CNS or lower respiratory tract of

sick premature new born infants
• M. hominis  Salpingitis PID , septic abortion, peritonitis

brain abscess
cause PAP similar to M. pneumoniae & meningitis in new born.
• Lab Diagnosis – M pneumoniae –
• Throat swab; Nasopharnygeal swab; Throat washings;
sputum BAL ,Genital Cervical swab
• Urethral swab
• Prostatic secretions; Tracheal aspirates; Urine
• Culture – PPLO broth
• Serological tests – Detection of antigen
• Immunfluroscence
• PCR
• Detection of Antibody - ELISA for IgM, IgG & IgA
• Cold agglutinins – M pneumonia – but Fp in Infectious
mononucleosis, influenza infections, cold
agglutinin disease, leukemia).
• Streptococcal MG test – Agglutination with
heat killed suspension of Streptococcus MG
titre of 1:20 is significant
• Treatment Since mycoplasmas lack a cell wall,

the penicillins and cephalosporins are
ineffective. The antibiotics of choice are
tetracycline (adults only) and erythromycin.
53. A veterinary doctor had pyrexia of unknown origin.
His blood culture in special laboratory media was
positive for gram negative short bacilli which was
oxidase positive. Which one of the following is the
likely organism grown in culture?
– Pasturella spp.
– Francisella spp.
– Bartonella spp.
– Brucella spp.
44. A farmer rearing sheep presented with complaints
of fever and weakness for lasting one month. There
is generalized LNP. There was also associated
hepatomegaly. Biopsy of liver showed non casting
granuloma – d/t
– Br. Mellitensie
– Br. Casis
– Francisella tularensis
– Yersinia pestis
• A man presents to em with hypotention ,
inguinal& axillary lymphadenopathy. Which
stains will be prefered for demonstration of
bipolar staining?
Classification
Disease Organism Vector Reservoir
Rocky Mountain R. rickettsii Tick Ticks, rodents

spotted fever
Rickettsialpox R. akari Mite Mites, rodents
Scrub typhus O. tsutsugamushi Mite Mites, rodents
Epidemic typhus R. prowazekii Louse Humans,

squirrel
fleas, flying
squirrels
Murine typhus R. thypi Flea Rodents
Q fever C. burnetii None Cattle, sheep,

(ticks in animals) goats, cats
Ehrlichiosis E. chaffeensis Tick Deer
E ewingii Tick Deer
• Most severe- Rocky mountain spotted fever
• Mild- R pox
• Eschar Seen-Scrub, R.pox, Indian tick typhus
• LN ↑Seen-Scrub, R.pox
• Vesicular /vericelliform rash- Rickettsial pox
• Transovarial transmission occurs only in tick & mite
• Rash – Epidemic (all over except palm & sole) Endemic (trunk>extremities)
RSM,ITT (palm sole) , No rash- Q fever
Systemic - Rocky mountain spotted fever
Lab diagnosis
• Isolation – in lab animals, hens egg & cell cultures
• Direct detection of the organisms & Bacteriology

7/5/2018
their antigens (staining) 445
Q Fever
• Infections occur worldwide
• The causative agent, Coxiella burnetii, is a C.burnetii
survives extreme environmental conditions for years;
very low infectious dose; transmitted by inhalation of
infectious aerosol
• Up to 60% of all cases are asymptomatic. Infection
may also occur by ingestion of infected milk or meat..
• No rash , No vector
Eschar+ LNpathy – R pox , scrub typhus

Agglutination
DISEASE OX19 OX2 OXK
Epidemic ++++ + _
typhus
Brill- +/ _ _ _
Zinsser
Endemic ++++ + _
typhus
Scrub _ _ +++
typhus
Spotted ++++ to+ +to++++ _
fever
Scrub typhus-
• Mite & chigger borne
• Affect rats in deserts, rain forest
• Eschar at site, rash, LN↑
• ↑OXK titre in Weil Felix
Coxiella burnetii –
• It survives pasteurization.
• Transmitted without arthropod (respiratory mode)
• No skin rash in Q fever.
• Zoonotic (wild animal)
• 7/5/2018
Produces interstitial pneumonia, shows latency
Bacteriology 449
• Ehrlichia  Small negative bacilli that multiply with
in membrane bound vacuoles in phagocytes
• Ehrlichiae laden vacuole is known as morula
• E sennetsu – illness resembling Glandular fever; pt

develops lymphoid hyperplasia & atypical
lymphocytosis seen in Japan & Malaysia. Suspected
to be ingested with raw fish
• E. chaffeensis – Infects human monocytic cells &

produce febrile illness. Transmitted to human by ticks
Multiple morulas of Ehrlichia canis in culture DH82 cells
Bartonellosis
• B.quintana and B.henselae are the most
important pathogens in this group of Gram-
negative rods
• B.bacilliformis: Oroya fever
• B.quintana- Trench fever (louse Born)
• B.henselae- cat-scratch disease (CSD) ,
Bacillary angiomatosis
• All can be important pathogens in AIDS and

transplant patients
• Donovanosis is best diagnosed by
– Microscopy
– Culture
– Skin testing
– Serology
• True abt scrub typhus are all except:
(AI2010)
A. Caused by tsutsugamushi
B. Tetracycline is DOC
C. Adults feed on host
D. Mite is the vector
• False about campylobacter is ?(aiims 2009)
a. Source of infec. Is poultary
b. Humans are the only reservoirs
c. Sequalae is GBS
d. skirrows media is used for culture.
The following is the etiological agent of Rocky
mountain spotted fever - (AIIMS May 05)
a.Rickettsia rickettsii
b.Rochalimaea quintana
c. Rickettsia tsutsugamushi
d.Coxiella burnetii

Scrub typhus us transmitted by -
(AIIMS Nov 07)
a.Reduvid bug
b.Trombiculid mite
c. Enteric pathogens
d.Cyclops

All of the following statement are true regarding
Q fever except - (AI 03)
a.It is zoonotic disease
b.Human disease is characterized by an interstitial
pneumonia
c. No rash is seen
d.Weil –felix reaction is very useful for diagnosis

• Family: Chlamydiaceae
• Genus: Chlamydia
• Species: trachomatis, pneumoniae, psittaci
Characteristics
• The Chlamydia is obligate intracellular parasites
(like virus) with a characteristic dimorphic growth cycle
• Different from virus

– Have DNA & RNA .
– Ribosomes , cell wall
– divided by binary fussion
– Susceptible to antibiotics.
• Different from true bacteria;

– have no peptidoglycon in their cell wall.
– Lack to ability to produce ATP.
Developmental cycle
Dev. cycle –
• elementary body (E/C, infectious form),
• reticulate body (I/C ,replicating)
CHLAMYDIA
• 4 species –
• (1) C. trachomatis HP)body
• (2) C. psittaci – (Levinthal Cole-Lillie bodies) (LCL)
• (3) C.pneumoniae, (atypical pneumonia ) TWAR agent
• (4) C.picorum
• C.trachomatis - 3 biovars – TRIC – A-K serovars
• A – C – blinding Trachoma ,
• D –K – Inclusion conjunctivitis, Ophthalmia neonatorum
Genital infection
• LGV – L1,L2,L3 serovars L2 - mc

• MC – ngu, oph neo,
• Obligate intracellular gram negative bacteria. Lack
peptidoglycan , c pneu- atherosclerosis
• Stain by casteneda , macchiauello, Giminez,

• Halbestaeder prowazek (HP)body in conjun. Epith. Cells
• Culture  Yolk sac, cell culture (McCoy Cells)

• Antigen detection – ELISA
• NAAT- PCR LCR
• Treatment  Tetracycline, EM, Sulfonamides
• For LGV  skin test  Frie test

– No genital lesion (only systemic manifestations)AIDS ,HBV,HCV
• LGV -
• MC by L2>L1,L3
• LGV serovars are more invasive than others
• MC LN involved- pararectal & intrapelvic nodes
• Pain less ulcer + painful LN↑
• Esthiomone (elephantiasis of vulva) rectal stricture
• Skin test  Frie test
• Inclusion body :
• LCL body- Psittacosis
• Miyagawa corpuscle- LGV
• HP body- trachoma

• Non gonococcal urethritis (NGU)
• 20-30% -Chlamydia trachomatis
• 10% - Mycoplasma genitalium & hominis
• Bacteroides urealyticus
• Herpes virus hominis
• Cytomegalovirus
• Trichomonas vaginalis
• 54. A patient complained of chills and fever
following a louse bite 2 weeks before. He had
rashes all over the body and was delirious at the
time of presentation to the hospital and
subsequently went into coma. A provisional
diagnosis of vasculitis due to Rickettsial infection
was made. Wyhich one of the following can be the
causative agent :
– Rickettsia typhi
– Rickettsia rickettiae
– Rickettsia prowazekii
– Ricketssia akari
52. Which of the following is not true about
Chlamydia
– Has biphasic life
– Reticulate body undergoes binary fission
– Elementary body is metabolically active
– Once it invades into cell it abates
phagolysosomal fusion
• 2. Noguchi medium is used for
– Bordettela
– Brucella
– Borrelia
– Corynebacterium
• A male patient presented with granulomatous
penile ulcer. On Wright giemsa stain tiny
organisms of 2 microns within macrophages
seen. What is the causative organism?
a) LGV
b) Calymmatobacterium granulomatis
c) Neisseria
d) Staph aureus
•
• DOC for chlamydia in prenancy ? (AI2010)
a. Tetra
b. Doxy
c. Erythro
d. penicillin
• Most sensitive tests of asymptomatic
chalmydia (AI2010)
a. Culture
b. NAAT
c. Serology
d. IFAT
• Haemophilus ducreyi is Causative agent of
– Hard chancre
– Soft chancre
– LGV
– Granuloma inguinale
True about pylori is-
A Vacuolating Cytotoxin
B Antiurease antibody detection specific for type I
C Found in the jejunum
D Treatment is Al(OH)
E It is flagellated
F. Involved in causation of peptic ulcer disease
G. Hypergastrinemia caused by it
H .It is a gram –ve organism
• Disease caused by Hemophilus - (PGI June 09)
a. Chancroid
b. Influenza
c. Acute epiglottitis
d. Brain abscess
e. Brazilian purpuric fever

Chlamydia except - (AIIMS May 05)
a.Erythomycin is effective for therapy of Chamydial
infections
b.Their cell wall lacks a peptidoglycan layer
c. They can grow in cell free culture media
d.They are obligate intracellular bacteria

Which one of the following statements is true
regarding Chlamydia pneumoniae - (AI 05)
a. Fifteen serovars have been identified as human
pathogens .
b.Mode of transmission is by the airborne bird excreta
c. The Cytoplasmic inclusions present in the sputum
specimen are rich in glycogen
d.The group specific antigen is responsible for the
production of complement fixing antibodies

The following statements are true with reference to
Mycoplasma except- (AIIMS Nov 05)
a. They are all smallest prokaryotic organisms that can
grow in cell free culture media
b.They are obligate intracellular organisms
c. They lack a cell wall
d.They are resistant to Beta-lactam drugs

True about Mycoplasma are all except -
(AIIMS May 08)
a.They are L forms
b.Sterol enhances growth
c. Can grow in cell free media
d.When grown in liquid medium do not produces
turbidity

40. Which is an obligatory intracellular
organisms
– Chlamydia
– Borrelia
– Actinomycetes
– Campylobacter
51. A male presents with profuse uretheral
discharge, on examination pus cells positive
but no organism is seen .For diagnosis you
would go for
– Culture on Mc-Coy cell
– Thayer-Martin media
– L.J. Medium
– Leventhal medium
Bacterial Vaginosis
• Caused by Gardnerella • Diagnosis- wet saline
vaginalis, Bacteroides, smear- Clue cells-
Mycoplasma hominis, epithelial cells
Ureaplasma surrounded by
urealyticum & coccobacilli all over the
Mobiluncus sps. surface.
• Creamy vaginal • Treatment with
discharge with fishy metronidazole.
smell
Clinical Features of Genital Ulcers
• Feature
• Syphilis - 1 endu,Firm, non tender, b/l non tender lnpathy
• Herpes- mult, no end, tender, b/l lnpathy -tender
• Chancroid – mult, soft, tender, U/l lnpathy tender

• Lymphogranuloma -1 firm, tender, U/l lnpathy venereum
• Donovanosis – Firm, pain Uncommon, No pain:
Pseudobuboes
A 35 year old patient complains of abdominal cramps along with profuse diarrhea.
The treating physician wants to process the stool specimen for isolation of
Campylobacter jejuni. Which of the following is the method of choice for the
culture of stool- (AIIMS Nov 04, AI 05)
a. Culture on TCBS medium incubated at 37°C under aerobic conditions
b. Culture on Skirrow’s medium incubated at 42°C under micro-aerophilic conditions.
c. Culture on MacConkey’s medium incubated at 42°C under anaerobic ocntions
d. Culture on Wilson and Blair’s medium incubated at 37°C under micro-aerophilic
conditions

• A man gets bite from a cat. The wound is
cleansed , and he received a booster of TT and
an injection of Penicillin G. After few days
wound is inflamed and sollen. exudates was
cultured on Blood agar and yielded Gram
negative rods. Most likely organism is
– Brucella canis
– Bartonella hensele
– Pasturella multilocida
– Clostridium tetani
• Bipolar staining, safety pin appearance
• Best stained with Wrights, Giemsa
• Donovan bodies  body of the bacillus blue &

capsule pink contained within the cytoplasmic
vacuoles of large macrophages
• Culture – Embryonated hens egg

• Treatment  tetracycline ,EM,chloramphenicol
316.A 25 year old farmer presented with history of high grade fever for
7 days and altered sensorium for 2days. On examination, he was
comatosed and had conjunctival hemorrhage. Urgent investigations
showed a hemoglobin of 11gm/dl, serum bilirubin 8 mg/dl and urea
78mg/dl. Peripheral blood smear was negative for malarial parasite.
What is the most likely diagnosis-
(AIIMS Nov 05)
a. Brucellosis
b. Weil’s disease
c. Acute viral hepatitis
d. Q fever
A fourteen year old body is admitted with history of fever,
icterus, conjunctival suffusion and haematuria for twenty
days. Which of the following serological test can be of
diagnostic utility- (AIIMS Nov 04)
a. Widal test
b. Microscopic Agglutination Test
c. Paul Bunnell test
d. Weil felix reaction

• The following are true regarding Lyme’s
Disease, except - (AI 03)
a. It is transmitted byixodid tick
b. Erythema chronicum migranies may be a clinical feature
c. Borrelia recurrentis is the aetiological agent
d. Rodents act as natural hosts

23. Which is not a test for syphilis
– TPHA
– Freis test
– Dark ground microscopy
– RPR
• 24. Most sensitive investigation for syphilis is
– FTA-ABS
– VDRL
– Wasserman’s reaction
– Kahn test
27. All are true about syphilis except
– VDRL becomes positive after 10-14 days of
chancre (3-5wk)
– The earliest serological test to appear in
untreated primary syphilis in TPHA
– Yaws & pinta cannot be differentiated
– VDRL titres  or even negative after RX
A VDRL reactive mother gave birth to an infant. All of the
following would help in determining the risk of transmission
to the infant except - (AI 06)
a. TPHA test on the serum sample of the mother
b. TPHA test on the serum sample of the infant
c. VDRL on the paired serum sample of the infant and mother
d. Time interval between the treatment of the mother and her
delivery

193.A 2 years old child is brought to the emergency with history of
fever and vomiting. On examination he has neck rigidity. The CSF
examination shows polymorphs more that 2000/ml protein 100mg
/dl and glucose 10mg/dl. The gram stain shows the presence of
Gram negative coccobacilli. The culture shows growth of bacteria
only on chocolate agar and not on blood agar. The causative agent
is - (AIIMS Nov 2002)
a. Neisseria meningitides
b. Haemophilus influenzae
c. Branhamella catarrhalis
d. Legionella pneumophila

• A 20 year old male patient present to the STD clinic with a
genital ulcer. The gram stain of the smear from ulcer shows
gram negative cocobacilli. The most appropriate media for
culture would be- (AI 04)
a. Thayer-Martin Medium
b. Blood agar with X & V factors
c. Chocolate agar with isovitate X
d. Tellurite blood agar

CALYMMATOBACTERIUM GRANULOMATIS
• Klebsiella granulomatis
• Produce granuloma inguinale, Granuloma
venerum or Donovanosis
• Clinical Forms
– Ulcerovegative - most common
– Hypertrophic or verrucous/ Necrotic/Cicatricial
• Painless red ulcer with fresh granulation tissue
•
GRANULOMA INGUINALE
• C/F  painless ulcer without lymphadenopathy
• Multiple ulcers of the vulva  destroy normal vulvar

architecture  scarring & lymphatic obstruction 
marked enlargement of the vulva
• Complications:
– Pseudobuboes, Elephantiasis, Squamous cell
carcinoma, Dissemination

Systemic Bacteriology

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Systemic Bacteriology

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Systemic Bacteriology

• Bacteria can be broadly divided into:

• Aerobic bacteria – bacteria which require O2

+ve CATALASE -ve

Bacilli/ Rods Cocci

1. Catalase positive – Staphylococcus species

MANNITOL S. Aureus : 1-3mm

Media for pigment

Glycerol monoacetate agar

• Tween agar, incubation at 22˚c

– Detects the presence of cytochrome oxidase enzyme.

• SLIDE COAGULASE TEST:

o Most strains of S. aureus have a bound coagulase enzyme

o This factor reacts with fibrinogen in plasma causing rapid

o Detects free coagulase which is

o Reacts with CRF in plasma to

o Has a unique ability to bind to Fc region of all human IgG

o Biological activities- interferes with opsonisation and ingestion

o Is the basis for coagglutination test , used in labs for org

o Absent in coagulase negative Staphylococci (CNS)

Alpha Haemolysin inactivated at 70C Chromosome S. aureus

Beta Haemolysin Neutral sphingomyelinase. Chromosome S. aureus

ENTEROTOXIN Heat stable responsible for Variable S. aureus

Rashdesquamation & Multi organ failure

Enterotoxins - Produced by 40% of clinical isolates

• (1961) - multi drug resistant

– important hospital pathogen

– Resistance is chromosomally mediated (mec A gene )

– Detection - 5g methicillin / 1g oxacillin disc

• - 4% NaCl, 30- 350C incubation

• Hosp acquired -(mecA I,II,III)

• Detection - using methicillin /oxacillin disc /cefoxitin disk

- on Screening agar media containing 4% NaCl

- incubation at lower temperature of 300 ,

- full 24hr incubation

– cipro/levofloxacin, cotrimoxazole, clindamycin, mupirocine,

– Newer – daptomycin, ceftobiprole, tigecycline, oritavancin

Disc Diffusion Method using Cefoxitin –

Using MHA incubated in ambient air @ 33-35 degC for 24 hrs

A} For S.aureus & S. lugdunensis

B} For CoNS-- R ≤ 24 mm ;S ≥ 25 mm,

• VISA - Vancomycin Intermediate Staphylococcus aureus -

• Mechanism – due to thickening of cell wall > VanA gene

• Treatment – same drugs given for MRSA except Vancomycin

• Skin & soft tissue infection

• S. saprophyticus - Coagulase negative, causes

B. PBP is essential for cell wall synthesis

C. PBP acts as carboxy peptidase & transpeptidase

D. alteration in PBP – basis of MRSA

Pike’s medium • Inhibiting the

Todd – •Used for luxuriant growth

1. Caps. hyaluronic acid • a) Synovial fluid

3. Erythema nodosum - Allergic reactions due to

 Antigenic & neutralized by convalescent sera.

Injected Into the skin of

Localized erythematous reaction appears

b) Streptococcal deoxyribonuclease: streptodorunase Four types A, B, C & D

e) Lipoproteinase (serum optical factor):

f) Other Extracellular products:

• They can be further divided into Enterococci & Nonenterococci

• resistant to penicillin, low level aminoglycosides & sulfonamides, cephalosporin,

• Combination therapy with penicillin & aminoglycoside is effective.

• Disease- UTI, endocarditis, intra abdominal infection, meningitis