Gram Posetive

MEDICAL IMPORTANCE BACTERIA
Common Pathogenic bacteria
3/10/2023 email : zenasahle@gmail,com 1

Learning objectives
– Describe pathogenic bacteria
– Describe
 virulence factors
 pathogenesis
 clinical diseases
 diagnosis pathogenic bacteria

Gram positive pathogens
• Species of interest
– Gram positive cocci
» Staphylococcus
» Streptococcus
» Enterococcus
– Gram positive bacilli
» Bacillus
» Clostridium
» Listeria

The genus staphylococcus
• The genus includes over 30 species and sub species
• S.aureus, S.epidermidis and S.saprophyticusfrequently
affect human host
General characteristics
• Gram positive cocci occurring in clusters
• Non-motile and non-sporing
• Some strains are capsulated
• Catalase positive
• Aerobes and facultative anaerobes
• Grows at wide temperature range (10-42℃)
• They are non fastidious and Grow on simple media
• They are small spherical organisms (1µm)
• B-lactamase production
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Characteristic differences
Characteristic S.aureus S.epidermidis S.saprophytic
us
Coagulase + - -
Deoxyribonucl + - -
ease
Novobiocin Susceptible Susceptible Resistant
Colonial Golden- Whitish Whitish
appearance yellow
Body sites Nose, Skin and Periurethra,

which may be mucosal mucosal feces
colonized surfaces, skin, surfaces
feces
S.aureus
Gram-positive spherical bacteria
– Clusters resembling grapes
– Produce yellow colony in culture
– Facultative anaerobes
– Catalase-positive
– Oxidase-negative
– Coagulase positive
– Can grow in temperature ranging from 15°C to 45°C)
Epidemiology
• S.aureus is ubiquitous organism can survive on dry surfaces for
long periods of time
• Transmission: Person-to-person spread through direct contact
or exposure to contaminated fomites (e.g., bed linens, clothing)
• Risk factors include hospitalization, previous surgical
procedure, and use of antibiotics that suppress the normal
microbial flora
S.aureus…
Pathogenesis
Depends on adherence, escape from phagocytosis and
tissue destruction through various virulence factors
Virulence factors
1. Structural components
– Capsule: Inhibits chemotaxis and phagocytosis
– Slime layer: Facilitates adherence to foreign bodies
– Peptidoglycan: stimulates production of endogenous pyrogen;
leukocyte chemo-attractant; inhibits phagocytosis
– Teichoic acid: Binds to fibronectin
– Protien A: Inhibits antibody-mediated clearance by binding
IgG1, IgG2, and IgG4 Fc receptors; leukocyte chemo-
attractant; anticomplementary
S.aureus…
2. Toxins
• Cytotoxins: Toxic for many cells, including
erythrocytes, fibroblasts, leukocytes, macrophages,
and platelets
• Exfoliative toxins (ETA, ETB): Serine proteases
that split the intercellular bridges in the stratum
granulosum epidermis
• Enterotoxins (A-R): Superantigens; stimulate
release of inflammatory mediators in mast cells
• Toxic shock syndrome toxin-1: Superantigen;
produces leakage or cellular destruction of
endothelial cells
3.Enzymes
A. Catalase
• Staphylococci produce catalase
– Converts H202 into H20 & 02
B. Coagulase
• Produced by S. aureus
• Extracellular protein which binds to prothrombin in the host to
form a complex called staphylothrombin
– It is reasonable to speculate that the bacteria could protect
themselves from phagocytic and immune defenses by causing
localized clotting

Cont…
Other enzymes
– Hyaluronidase: to facilitate spread of S.
aureus in tissue
– Lipase: associated with superficial skin
infection
– Nuclease: produced only by S. aureus
– Penicillinase (disrupts the Beta-lactame rings)

staphylococcal disease
Toxin-mediated Diseases
• Scalded skin syndrome (Ritter disease):
Disseminated desquamation of skin epithelium
especially in infants; blisters with no organisms or
leukocytes.
• Food poisoning: After consumption of food
contaminated with heat-stable enterotoxin, rapid onset
of severe vomiting, diarrhea, and abdominal
cramping, with resolution within 24 hours
• Toxic shock: multisystem intoxication characterized
initially by fever, hypotension, and a diffuse, macular
erythematous rash; high mortality without prompt
antibiotic therapy and elimination of the focus of
infection
staphylococcal disease…
Scalded skin syndrome Toxic shock syndrome

staphylococcal disease …
Suppurative Infections
• Impetigo: localized cutaneous infection characterized by
pus-filled vesicle on an erythematous base
• Folliculitis: impetigo involving hair follicles
• Furuncles or boils: large, painful, pus-filled cutaneous
nodules
• Carbuncles: coalescence of furuncles with extension
into the subcutaneous tissues and evidence of systemic
disease (fever, chills, bacteremia)
• Bacteremia and endocarditis: spread of bacteria into
the blood from a focus of infection; endocarditis
characterized by damage to the endothelial lining of the
heart
staphylococcal disease …
Furuncles
Pustular
impetigo
Curbuncle
• Pneumonia and empyema:
Consolidation and abscess formation in the lungs
Seen in the very young and elderly and in patients with
underlying or recent pulmonary disease; a severe form
of necrotizing pneumonia with septic shock and high
mortality is recognized.
• Osteomyelitis:
Destruction of bones, particularly the metaphyseal area
of long bones
• Septic arthritis:
Painful erythematous joint with collection of purulent
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Disease caused by CoNS and possibly S. aureus
• Wound infections:
Characterized by erythema and pus at the site of a traumatic
or surgical wound
• Urinary tract infections:

Dysuria and pyuria in young sexually active women
(Staphylococcus saprophyticus), in patients with urinary
catheters (other CoNS), or following seeding of the urinary
tract by bacteremia (S. aureus)
• Catheter and shunt infections:

Chronic inflammatory response to bacteria coating a
catheter or shunt (commonly with CoNS)
• Prosthetic device infections:

Chronic infection of artificial joint characterized by
localized pain and mechanical failure of the joint (most
commonly with CoNS)

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Pathogenesis of staphylococcal infections
Laboratory diagnosis
• Microscopy: for Pyogenic infections
• Culture: staphylococci can readily grow in
Nutrient agar and Blood agar; Mannitol-salt
agar is selective for S.aureus
• Biochemical test: Catalase, Coagulase,
DNAase test
• Nucleic acid amplification tests especially to
detect MRSA
Gram stain showing Staphylococcus S.aureus colonies on

3/10/2023GPC in clusters colonies
email in blood agar
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Treatment and Prevention
• Oral therapy can in include trimethoprim-sulfamethoxazole,
doxycycline or minocycline, clindamycin, or linezolid
• Vancomycin is drug of choice for intravenous therapy, with
daptomycin, tigecycline or linezolid acceptable alternatives
• Treatment is symptomatic for patients with food poisoning
• Proper cleansing of wounds and use of disinfectant
• Hand washing and covering of exposed skin
• Removal of the foreign body (medical devices)
• Prompt treatment for endocarditis or shunt infections is
necessary to prevent further tissue damage or immune complex
formation

Genus Streptococcus & Enterococcus
General characteristics
• Round to oval gram positive cocci arranged in pairs or chains
• Non-motile, non-sporing; Fastidious organisms
• Mostly facultative anaerobes
• Catalase negative unlike staphylococci
Classification is based on 3 Schemes
1. Serologic properties (Lancefield grouping)
– Grouped as A,B,D etc based on the polymeric carbohydrate in their cell
walls
2. Patterns of heamolysis on blood gar
 (Complete hemolysis (β-hemolytic)
 Partial hemolysis (α-hemolytic)
 Non-hemolytic (γ-hemolytic)
3. Biochemical reaction (physiologic properties): eg salt tolerance

Streptococcus pyogens
• Rapidly growing gram-positive cocci arranged in

chains;
• B-hemolytic colonies on blood agar
• Contains group-specific carbohydrate (A antigen) and
type-specific proteins (M protein) in its cell wall
Epidemiology
• S. pyogenes is an upper respiratory tract commensal
• Infection is the result of Person-to-person spread by
respiratory droplets (pharyngitis) or through breaks in
skin after direct contact with infected person, fomite,
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Streptococcus pyogens…
Pathogenesis
• Determined by the ability of the bacteria to avoid opsonization
and phagocytosis, adhere to and invade host cells, and produce
a variety of toxins and enzymes.
Virulence factors
• Capsule: poor immunogen &interferes with phagocytosis
• Fimbriae/pili, F-protien, M protien and LTA: facilitate
adherence
• M protien: blocks the binding of the complement (C3b)
• M-like proteins: bind Abs; block phagocytosis and activation
of complement

• Streptolysins (haemolysins): streptolysins O and
S lyse erythrocytes and are cytotoxic to leukocytes
and other cell types.
• C5a-peptidase: inactivates C5a (complement) and
reduces chemotaxis
• Streptokinase: Prevents formation of fibrin mesh
• Deoxyribonuclease, Hyaluronidase
• Streptococcal pyrogenic exotoxins (erythrogenic
toxins): responsible for the rash of scarlet fever
– Are superantigensemail
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potentially leading to shock 24
Clinical disease
Suppurtive disease
• Scarlet fever: diffuse erythematous rash beginning on the chest
and spreading to the extremities; complication of streptococcal
pharyngitis
• Pyoderma: localized skin infection with vesicles progressing to
pustules; no evidence of systemic disease
• Erysipelas: localized skin infection with pain, inflammation,
lymph node enlargement and systemic symptoms
• Cellulitis: infection of the skin that involves the subcutaneous
tissues
• Streptococcal toxic shock syndrome: multiorgan systemic
infection resembling staphylococcal toxic shock syndrome;
however, most patients are bacteremic and with evidence of
fasciitis
S. pyogens
Clinical disease
Necrotizing fasciitis: deep infection of skin that involves
destruction of muscle and fat layers
Acute stage erysipelas

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• Pharyngitis: reddened pharynx with exudates; cervical
lymphadenopathy can be prominent
• Sinusitis, tonsillitis, otitis media, pneumonia
• Bacteremia, Septic arthritis
• Genitourinary: puerperal sepsis (infection of female
reproductive tract)
• Cardiovascular: infective endocarditis
Non supurative infections
• Rheumatic fever: characterized by inflammatory
changes of the heart (pancarditis), joints (arthralgias to
arthritis), blood vessels, and subcutaneous tissues
• Acute glomerulonephritis: acute inflammation of the
renal glomeruli with edema, hypertension, hematuria,
and proteinuria
Pathogenesis of S pyogenes
infections.
• Microscopy: gram stain of affected tissues help

the dx of S. pyogenes soft tissue infections or
pyoderma.
• Antigen detection from throat swab
• Antibody detection: ASO test: for confirming

rheumatic fever and Acute glomerulonephritis
• Nucleic acid amplification tests: for pharyngeal

Lab diagnosis
• Culture in Blood agar or Chocolate agar (helpful in case
of impetigo, necrotizing fasciitis; however, relatively
few organisms may be present in the skin of patients
with erysipelas or cellulitis)
• Bacitracin susceptibility, L-pyrrolidonyl arylamidase
(PYR) test
• Serology: ASO titer,
Prevention and control
• Treatment: oral penicillin, amoxacillin; oral
cephalosporin or macrolide for pts allergic to penicillin
• Combined treatment of penicillin + clindamycin for
sever systemic infections
• Surgical debridement
Streptococcus agalactiae (GBS)
• S.agalactiae is Lance field group B

• B-hemolytic colonies on blood agar; Gram positive cocci
in chain
• Part of the normal flora faecal, perineal and vaginal flora
in females
• Transmission: Early-onset disease acquired by neonates
from mother during pregnancy or at time of birth
• Risk groups
Neonates, if there is premature rupture of membranes,
prolonged labor, preterm birth, or disseminated maternal group
B streptococcal disease
Streptococcus agalactiae (GBS)
 Risk groups
Mother is without type-specific antibodies and has low
complement levels; Women with genital colonization
are at risk for postpartum disease
Men and non pregnant women with diabetes mellitus,
cancer, or alcoholism are at increased risk for disease
Virulence factors
• Polysacharide capsule: interferes with phagocytosis
• Sialic acid (terminal residues of polysacharide capsule):
inhibits activation of alternative complement
pathway
S.agalactiae…
Clinical disease
• Respiratory tract: pneumonia in neonates and the elderly
• Musculoskeletal: septic arthritis, osteomyelitis
• Skin and soft tissue: cellulitis and wound infections
• Systemic: Bacteremia
• Genitourinary: In the post-partum period it causes septic
abortion, endometritis and urinary tract infections
• Cardiovascular: infective endocarditis
• Central nervous system: neonatal meningitis

S.agalactiae…
• Microscopy (gram stain): useful for meningitis

(CSF), pneumonia (lower respiratory secretions),
and wound infections (exudates)
• Culture: most sensitive test
• Polymerase chain reaction–based assays

S.agalactiae…
• Penicillin G is the drug of choice
• Empiric therapy: cephalosporin plus
aminoglycoside; combination of penicillin and
aminoglycoside is used in patients with serious
infections; a cephalosporin or vancomycin is used
for patients allergic to penicillin
For high-risk babies, penicillin is given at least
4 hours before delivery
All pregnant women should be screened for
colonization with group B streptococci at 35 to 37
weeks of gestation
Chemoprophylaxis of risk groups
S. pneumoniae (Pneumococcus)
Distinctive features
• Gram positive cocci commonly found in pairs
• Grow in blood agar or chocolate agar with C2O
• α-Hemolytic, centrally depressed colonies
• Capsulated with polysacharide capsule
• Sensitive to optochin and bile salts
Epidemiology
• S. pneumoniae is a commensal of the upper respiratory tract
• Most infections are caused by endogenous spread from the
colonized nasopharynx or oropharynx to distal site
• Person-to person spread through infectious droplets is rare
• Disease is more common in cool months
S.pneumoniae…
Pathogenesis and virulence
• Surface protein adheisn: enable colonization to oropharynx
• Pneumolysin: damage ciliated epithelial cells and
macrophages
• IgA protease: cleave secretory IgA
• Polysaccharide capsule: Inhibit phagocytosis
• Tiechoic acid, cell wall fragments and pneumolyisn
stimulate local inflammatory response
Clinical disease
• Respiratory infections
– Otitis media, sinusitis, lower respiratory tract infection (particularly
community-acquired pneumonia);
• Systemic infection: Bacteremia (especially in 25-30% of
pneumococcal pneumonia)
• The bacteria also causes Meningitis
Pathogenesis of S pneumoniae
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S.pneumoniae
• Microscopy: Gram stain of sputum for pneumonia
&meningitis; Quellung reaction to reveal the capsule
• Antigen detection from CSF
• Culture on sheep blood agar
• Nucleic acid probes and PCR assays for isolates from
culture
• Biochemical test: S.pneumoniae is bile soluble and
Opthochin sensitive unlike viridian streptococci
Treatment, Prevention and control
• Treatment: Penicillin, vancomycin±𝑐𝑒𝑓𝑡𝑟𝑖𝑎𝑥𝑜𝑛𝑒,
Cephalosporins and flouroquinolones
• Vaccination: 13-valent conjugated vaccine (children < 2
yrs); 23-vallent polysaccharide vaccine (for adults at
risk)
Viridian streptococci
• The ‘viridans’ streptococci are a group of α- or non
haemolytic streptococci
• Predominantly found in the oral cavity and are
commonly referred to as the ‘oral’ streptococci.
• Examples include
– Streptococcus mitis, Streptococcus mutans, Streptococcus
salivarius and Streptococcus sanguinis groups
Epidemiology
• They are commensals of the upper respiratory tract.
Pathogenesis
• Various carbohydrates facilitate the attachment of
these streptococci to teeth adjacent to the gingivae
and form biofilms (dental plaque)
Viridian streptococci…
 The bacteria use dietary sugar and form lactic acid which
damages the tooth enamel. Some species, particularly
S.mutans, produce acid involved in the development of
dental caries.
Associated infections
• Cardiovascular infections : they are common (50-70%)
cause of infective endocarditis;
• Dental disease: these streptococci, particularly S.mutans
are the most common cause of dental caries and
periodontal disease.
Viridian streptococci
Lab diagnosis
• Culture: isolation from infected sites
• Biochemical test: the bacteria are resistant to optochin
and insoluble in bile salts
Treatment
• Viridian streptococci are sensitive to wide variety of
antibiotics
• Often treated with penicillin
• Prophylaxis may be given before dental procedures
Genus Enterococcus…
• Previously classified as group D streptococci
• Currently contains 40 species but Enterococcus faecalis and
Enterococcus faecium are mostly commonly associated with
human infection
• Gram-positive cocci arranged in pairs or chains
• Facultative anaerobic; non-sporing , non-motile (except
Enterococcus casseliflavus and Enterococcus gallinarum);
• On blood agar they appear α-, β- or sometimes non-
haemolytic
• Some strains are encapsulated
• Can grow over wide temperature range.

Enterococcus…
Epidemiology
• Enterococci are commensals, most notably of the
gastrointestinal and vaginal tract
• Cell wall structure allows survival on
environmental surfaces for prolonged periods
• Most infections are endogenous; some caused by
patient-to-patient spread
• Patients at increased risk include those
hospitalized for prolonged periods and treated
with broad-spectrum antibiotics (particularly
cephalosporins, to which enterococci are
naturally resistant)
Enterococcus…
Virulence and pathogenesis
• Virulence mediated by ability to adhere to host
surfaces and form biofilms and by antibiotic resistance
• Enterococcal strains can produce cytolysin, which
causes lysis of a variety of cells, including erythrocytes
and other mammalian cells.
• Gelatinase, hyaluronidase, aggregation substance,
Enterococcal Surface Protein and extracellular
superoxide production
Enterococcus…
Clinical disease
• Urinary tract infection: dysuria and pyuria most commonly in
hospitalized patients with an indwelling urinary catheter and
receiving broad-spectrum cephalosporin antibiotics
• Peritonitis: abdominal swelling and tenderness after abdominal
trauma or surgery; patients are typically acutely ill, febrile, and
with positive blood cultures; typically polymicrobic infection
• Bacteremia: associated with either a localized infection or
endocarditis
• Endocarditis: infection of the heart endothelium or valves;
associated with persistent bacteremia; can present acutely or
chronically

Enterococcus…
Lab diagnosis
• Culture: grows readily on Blood agar, Chocolate agar, BHI agar etc
• Biochemical test: Enterococci are Catalase negative, L-pyrrolidonyl
arylamidase–positive, resistant to bile and optochin; grow in media
containing 6.5% NaCl.
Treatment
• Enterococcal infections are often treated with ampicillin.
• Enterococci, (particularly E. faecium), have developed resistance to
penicillin
• The emergence of resistance to glycopeptide antibiotics such as
vancomycin is of concern
Gram positive bacilli
• Bacillus
• Clostridium
• Listeria

Gram positive bacilli

Genus bacillus
• Contains almost 250 species

• Bacillus anthracis and Bacillus cereus are species
of medical interest
B. anthracis
Morphology
• Spore-forming, nonmotile, nonhemolytic gram-
positive rods
• Polypeptide capsule consisting of poly-D-
glutamic acid observed in clinical specimens

B.anthracis…
Epidemiology
• Anthrax is primarily a disease of herbivores;
• humans are infected through exposure to contaminated
animals or animal products.
Human B. anthracis disease is acquired through:
• Inoculation: Up to 95% anthrax infections are due to
inoculation of Bacillus spores through exposed skin
from either contaminated soil or infected animal
products, such as hides, goat hair, and wool.
• Ingestion of spores: Very rare in humans, but ingestion
is a common route of infection in herbivores.
• Inhalation of spores: The most likely route of infection
with biologic weapons
• No person to person spread
B.anthracis…

Bacillus anthracis …
Bacillus anthracis as biological weapon

B.anthracis…
Pathogenesis
• Virulence factors include:
Capsule
Edema toxin: Increase intracellular cAMP and results in
accumulation of fluid (edema)
Lethal toxin: Stimulates macrophages to release TNF-α,
interleukin-1β, and other proinflammatory cytokines and
responsible for tissue necrosis.
Protein antigens: mediate entry of edema factor and
lethal toxin to the cell
B.anthracis…

B.anthracis…

B.anthracis…
Clinical disease
• Cutaneous anthrax
– Painless papule progresses to ulceration with surrounding
vesicles and then to black eschar formation; painful
lymphadenopathy, edema, and systemic signs may develop
• Gastrointestinal anthrax
– ulcers form at site of invasion (e.g., mouth, esophagus, intestine)
leading to regional lymphadenopathy, edema, and sepsis
• Inhalation anthrax
– Initial nonspecific signs followed by the rapid onset of sepsis
with fever, edema, and lymphadenopathy (mediastinal lymph
nodes); meningeal symptoms in half the patients, and most
patients with inhalation anthrax will die unless treatment is
initiated immediately

B.anthracis…

B.anthracis…
Lab diagnosis
CAUTION: Laboratory safety is very
important when working with any materials
suspected of containing Bacillus anthracis
– Gram stain from wound or blood Gram stain of Bacillus
specimens: Seen as long thin gram anthracis showing Gram-
positive rods singly or in chains positive rods
– Spore stain (for isolates from culture):

Malachite green stain
– Capsule: Indian ink, M’Fadyean
methylene blue stain, or a direct
fluorescent antibody (DFA) test
• Culture: non pigmented, non hemolytic Colonial growth of
colonies on sheep blood agar. “Medusa Bacillus anthracis on
head” morphology on nutrient
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sheep blood agar
59
Bacillus cereus
Characteristic
• Spore-forming, motile gram-positive rods
• Hemolytic on blood agar
• Can grow under anaerobic conditions
Epidemiology
• Ubiquitous in soils throughout the world
• People at risk include
– Those who consume food contaminated with the bacterium
(e.g., rice, meat, vegetables, sauces)
– Those with penetrating injuries (e.g., to eye), those who
receive intravenous injections, and immunocompromised
patients exposed to B. cereus
Bacillus cereus…
Pathogenesis
B. cereus produces two type of enterotoxins
• Heat-stable enterotoxin: causes the emetic
form of the disease and
• heat-labile enterotoxin: causes the diarrheal
form of the disease
• Necrotic toxin, cereolysin and phospholipase
C cause tissue destruction in eye infections
Clinical disease
• Gastroenteritis: emetic form characterized by
a rapid onset of vomiting and abdominal pain
Bacillus cereus…
Ocular infections: rapid, progressive

destruction of the eye after traumatic
introduction of the bacteria into the eye
Severe pulmonary disease: severe anthrax-

like pulmonary disease in immunocompetent
patients
Lab Diagnosis
Genus Clostridium
• Large gram positive bacilli
• Strictly anaerobic, spore-forming, fermentative
organisms
• Catalase negative
• Clostridia are widely distributed in the
environment and in the gastrointestinal tract of
mammals
• Medically important species include
• C. perfingens,
• C. difficile,
• C. tetani,
• C. botulinum, C.
3/10/2023 emailsepticum
: zenasahle@gmail,comand C. tertium 63
C.perfringens…
Morphology and identification
• Non-motile, sub-terminal spores.
• Forms irregular, spreading colonies on blood agar surrounded
by a double zone of B-haemolysis
• Five types (A-E) of C.perfringes based on surface antigen and
type of toxin produced
• Type A strains: commonly found in human infections; produce
only a-toxin
• Type B to E strains: commonly found in animals produce a-
and other toxins
Pathogenesis
• Enzyme & toxin production: α-toxin, Hyaluronidase,
Collagenase & lipase ???
C.perfringens…
Clinical signs of gas gangrene

• Pain, edema
• Bloody exudate in the lesion
• Fever
• Tachycardia
• Blackened necrotic tissue filled with bubbles of gas
Other infections
• Anaerobic cellulitis
• Endometritis
• Food poisoning: Nausea, abdominal pain, and diarrhea
8-24 hrs after ingestion of spore contaminated food
C.perfringens
Lab diagnosis
• Disease is diagnosed clinically lab findings are
supportive
• Gram stain of pus, blood or tissue
• Culture: need anaerobic environment
• alpha-toxin and lipase production in egg-yolk agar
• Surgical removal of all the dead and foreign matter and
immediate antibiotic therapy with high dose
benzylpenicillin and/or metronidazole and supportive
measures
• Food poisoning is self limiting
• Good cooking hygiene
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and adequate refrigeration
C.tetani
Morphology and identification
• Terminal spore (‘drumstick’ appearance);
• Produces a thin spreading film of growth without discrete
colonies on blood agar;
• Motile with peritrichous flagella.
Epidemiology
• C. tetani is present in mammalian intestines and the
environment (particularly manured soil).
• The spores are introduced into wounds contaminated
with soil or foreign bodies

C.tetani…
Pathogenesis
• Many strains are highly toxigenic, producing
oxygen-labile haemolysin (tetanolysin) and a
potent neurotoxin (tetanospasmin).
• Tetanospasmin produced at the local site of
infection ascends through nerves to anterior horn
Tetanospasmin blocks neurotransmitter release, resulting
in the characteristic motor spasms

C.tetani…
Clinical disease
• C.tetani causes tetanus
• Incubation period: 4-10 days
• Early symptom is trismus (lock jaw):
Spasms of the masseter muscle
Difficulty in opening of the mouth and masticating
Rigidity spreads to muscles of the face, neck and
truck
• Back is usually slightly curved (Opisthonotus )
• The spasms gradually intensify and patient may die of
Paralysis of respiratory muscles and respiratory
collapse
• Fatality rate is 10-70%
C.tetani…
Opisthotono
Trismus s
C.tetani
Diagnosis is more of clinical
• Lab diagnosis
– Anaerobic culture
– Serological detection of circulating neurotoxin by enzyme
immunoassay
Treatment
• Neutralization of toxin with human tetanus immune
globulin
• Benzodiazepines: antagonize the toxin
• Supportive measures: dark environment, airway,
sedation
Prevention
• Childhood toxoid immunization prevents disease
• Boosters required every 10 years
• Passive immunity with
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a prophylactic dose of HTIG
C. Butulinum
• Oval, sub-terminal endospores
• Motile (perithricous flgella)
• Spore, relatively heat resistant (survive improper canning

and growth encouraged in anaerobic environment)
• Spore present in decaying vegetation, intestinal tract of

birds, mammals, sedments of lakes, ponds, soil.
• Spores of the organism are highly resistant to heat,

withstanding 100 °C for several hours.
Virulence factors
• Botulism toxin/ neurotoxin
– it is heat labile
• Seven toxin types (A-G)/serotypes
– Human illness is caused by mainly Type A, B, & E, and
rarely by F
– Type E linked with fish products
– Type A & B associated with variety of foods
– Type D cause botulism in mammals ----- important!!!
– Types C and D are encoded by bacteriophage that infect
the bacteria.
• Affect mainly peripheral nervous system
– Prefer stimulatory motor neurons
– Weak/ flaccid paralysis
• The toxin is formed in food when C. botulinum spores
contaminate food.
Pathogenesis
Food-borne Botulism
• Botulism is not infection but an intoxication resulting from the
ingestion of food in which C. botulism has grown (spores have
germinated) & produced toxin
• rare and usually fatal
• Result from an ingestion of a lethal preformed neurotoxin, about
18-36 hours following ingestion of preformed toxin,
• These protein exotoxins are often released in an inactive form,
then proteolytic cleavage activates them.

Clinical findings
• Sign & symptoms begin: 18 - 36 hrs after ingestion of toxic food
• Visual disturbances (blurred vision)
• Difficulty to swallow
• Speech difficulty
• Gastrointestinal symptoms are not prominent
• No fever, dizziness and dryness of the mouth
• Descending weakness of skeletal muscles
• Death occurs from respiratory paralysis or cardiac arrest
• The patient remains fully conscious until shortly before death.
• In Infant botulism
• constipation, weak sucking ability, generalized weakness
occur in infants at 5-20wks of age

• Spores of C. botulinum are widely distributed in soil,
• They often contaminate vegetables fruits and other foods
• When such foods are canned / preserved, they either:
• must be sufficiently heated to destroy spores or
• boiled for 20min before consumption
• Strict regulation of commercial canning
• Careful observation of the cans
– E.g. Swelling of the cans
Treatment
• Trivalent (A, B, E,) anti - toxins are available
• Promptly administered intravenously
Clostridium difficile
• Slender bacilli - with large, oval, subterminal spores
• Members of the intestinal flora
• Nosocomial pathogen
Virulence factor
• Two toxin: Toxin A (enterotoxin), Toxin B (extremely lethal/cytopathic
toxin)
Pathogenesis
Pseudomembranous colitis
– Although many antibiotics have been associated with
pseudomembranous colitis, the most common are
ampicillin and clindamycin.
– Administration of antibiotics results in proliferation of
drug-resistant C difficile that produces two toxins.
• Toxin A, a potent enterotoxin that also has some cytotoxic
activity, binds to the brush border membranes of the gut at
receptor sites.
• Toxin B is a potent cytotoxin.
Antibiotic-Associated Diarrhea
– The administration of antibiotics frequently leads to a mild to
moderate form of diarrhea, termed antibiotic-associated diarrhea.
– This disease is generally less severe than the classic form of
pseudomembranous colitis.
– As many as 25% of cases of antibiotic-associated diarrhea may be
associated with C difficile.
– The diarrhea may be watery or bloody, and the patient frequently has
associated abdominal cramps, leukocytosis, and fever
Lab diagnosis
• detection of one or both C difficile toxins in stool
• endoscopic observation of pseudomembranes or
microabscesses in patients who have diarrhea and have
been given antibiotics.
Treatment
• The disease is treated by discontinuing administration of
the offending antibiotic and orally giving either
metronidazole or vancomycin.
L. monocytogens
General characteristics:
• Widely present in plants, soil and surface water
• Zoonotic pathogen of domestic animals
• Non-sporulating, facultative anaerobe, intracellular.

Gram positive rods
Antigenic structure:
• . Listreriolysin( hemolysin)

Pathogenesis and clinical features:
• Transmitted to humans through ingestion of poorly coooked meat

and unpasteurized milk and milk products
1. Perinatal human listeriosis: Granulomatous infantisepticum
– . Early onset syndrome: Intrauterine sepsis
– . Late onset syndrome: Neonatal meningitis

2. Adult human listeriosis:
– . Meningoencephalitis
– . Bacteremia
Lab. Diagnosis:
• Specimen: Blood/ CSF
• Culture: Grow in blood agar and demonstrate
narrow zone of β-hemolysis
• Biochemical reaction:
– Catalase positive
– Oxidase negative
Treatment:
• . Ampicillin
• . Erythromycin
• . Cotrimaxazole
Prevention and control:
• . Proper cooking of animal souce foods
• . Pasteurization of milk and milk products
Any questions?

Gram Posetive

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MEDICAL IMPORTANCE BACTERIA

Common Pathogenic bacteria

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Body sites Nose, Skin and Periurethra,

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Scalded skin syndrome Toxic shock syndrome

• Urinary tract infections:

• Catheter and shunt infections:

• Prosthetic device infections:

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Gram stain showing Staphylococcus S.aureus colonies on

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• Rapidly growing gram-positive cocci arranged in

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Acute stage erysipelas

• Microscopy: gram stain of affected tissues help

• Antigen detection from throat swab

• Antibody detection: ASO test: for confirming

• Nucleic acid amplification tests: for pharyngeal

• S.agalactiae is Lance field group B

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• Microscopy (gram stain): useful for meningitis

• Culture: most sensitive test

• Polymerase chain reaction–based assays

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• Contains almost 250 species

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Bacillus anthracis as biological weapon

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– Spore stain (for isolates from culture):

Ocular infections: rapid, progressive

Severe pulmonary disease: severe anthrax-

Clinical signs of gas gangrene

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• Motile (perithricous flgella)

• Spore, relatively heat resistant (survive improper canning

• Spore present in decaying vegetation, intestinal tract of

• Spores of the organism are highly resistant to heat,

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• Widely present in plants, soil and surface water

• Zoonotic pathogen of domestic animals

• Non-sporulating, facultative anaerobe, intracellular.

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• Transmitted to humans through ingestion of poorly coooked meat

1. Perinatal human listeriosis: Granulomatous infantisepticum

– . Early onset syndrome: Intrauterine sepsis

– . Late onset syndrome: Neonatal meningitis

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