Aerobic bacilli (genus:
Aerobic bacilli (genus:
B
B
acillus)
acillus)
G
G
ram-negative oxidase-
ram-negative oxidase-
positive microaerophilic
positive microaerophilic
fermenting rods (genera:
fermenting rods (genera:
C
C
ampylobacter and
ampylobacter and
H
H
elicobacter)
elicobacter)
Bacillus spp.
Bacillus spp.
Aerobic spore-forming
Aerobic spore-forming
bacteria pathogenic for
bacteria pathogenic for
humans
humans
B. anthracis
B. anthracis
B. cereus
B. cereus
B. subtilis
B. subtilis
General characteristics
General characteristics
o
o
f
f
B
B
.
.
a
a
nthracis
nthracis
Gram
Gram
-positive
-positive
R
R
ods creating short
ods creating short
chains or pairs
chains or pairs
N
N
onmotile
onmotile
Spores in the central part
Spores in the central part
of cell
of cell
In living organism creates
In living organism creates
big capsule (!)
big capsule (!)
Production of pathogenic
Production of pathogenic
toxin
toxin
Existence in soil
Existence in soil
Capsule of
Capsule of
B
B
.
.
a
a
nthracis
nthracis
Contains a virulence factor
Contains a virulence factor
Encoded in bacterial plasmids
Encoded in bacterial plasmids
(only these strains are pathogenic)
(only these strains are pathogenic)
Built from
Built from
polypeptides
polypeptides
Pr
Pr
otects against phagocytosis
otects against phagocytosis
B.
B.
a
a
nthracis
nthracis
toxin
toxin
Encoded in bacterial plasmids
Encoded in bacterial plasmids
Consists of: protective antigen,
Consists of: protective antigen,
lethal factor and edema factor
lethal factor and edema factor
A
A
ffects the central nervous system
ffects the central nervous system
Is a v
Is a v
irulence factor of
irulence factor of
B. anthracis
B. anthracis
Anthrax (zoonotic
Anthrax (zoonotic
disease)
disease)
IN ANIMALS
IN ANIMALS
Digested with
Digested with
grass
grass
Inhaled with air
Inhaled with air
Passed through
Passed through
wounded skin
wounded skin
IN HUMANS
IN HUMANS
Foodborn
Foodborn
e
e
infections
infections
Airborn
Airborn
e
e
infections
infections
Through abrasions
Through abrasions
on the skin
on the skin
routes of infection - spores or
vegetative forms
Clinical manifestation of
Clinical manifestation of
anthrax
anthrax
Pulmonary anthrax (the severest form)
Pulmonary anthrax (the severest form)
:
:
Gastrointestinal anthrax
Gastrointestinal anthrax
Cutaneous anthrax (most common form)
Cutaneous anthrax (most common form)
Septic
Septic
a
a
emia (after passing bacteria into the
emia (after passing bacteria into the
bloodstream)
bloodstream)
Materials for examination
Materials for examination
depen
depen
d
d
on clinical
on clinical
symptom
symptom
s: s
s: s
putum
putum
, s
, s
mear from the skin
mear from the skin
lesion
lesion
, b
, b
lood
lood
Bacterial examination
Bacterial examination
Microscopic view
Microscopic view
in material
in material
s:
s:
capsules
capsules
are seen
are seen
Slide (Gram or Ziehl - Neelsen –staining)
Slide (Gram or Ziehl - Neelsen –staining)
Routine bacteriological media (nutrient
Routine bacteriological media (nutrient
broth, nutrient agar)
broth, nutrient agar)
, a
, a
erobic conditions
erobic conditions
Colony: big, colorless, rough
Colony: big, colorless, rough
Microscopic view
Microscopic view
after culturing
after culturing
: spores
: spores
in the central part of cell
in the central part of cell
Biochemical tests
Biochemical tests
, b
, b
iological assay (white
iological assay (white
mice)
mice)
Antibiotic therapy
Antibiotic therapy
of anthrax
of anthrax
Penicillin
Penicillin
Cephalosporines
Cephalosporines
Chloramphenicol
Chloramphenicol
Aminoglycosides
Aminoglycosides
Effective
Effective
only
only
to cutaneous anthrax
to cutaneous anthrax
and in early systemic diseases
and in early systemic diseases
B
B
acillus
acillus
cereus
cereus
Motile bacteria (unlike
Motile bacteria (unlike
B
B
.
.
anthracis
anthracis
)
)
Common in the environment
Common in the environment
Causes food poisoning called
Causes food poisoning called
gastroenteritis
gastroenteritis
Severe infections in
Severe infections in
immunosuppressed people
immunosuppressed people
Campylobacter
Campylobacter
spp.
spp.
C. jejuni
C. jejuni
C. coli
C. coli
C. fetus
C. fetus
C. sputorum
C. sputorum
C. consisus
C. consisus
Zoonotic bacteria
Zoonotic bacteria
transmitted
transmitted
by f
by f
a
a
ecal-
ecal-
oral
oral
route
route
(water, food)
(water, food)
C
C
haracteristics of
haracteristics of
Campylobacter
Campylobacter
spp.
spp.
Gram
Gram
-negative
-negative
C
C
urved rods in pairs
urved rods in pairs
(“gull wing” after
(“gull wing” after
Gram staining)
Gram staining)
R
R
equire 10% CO
equire 10% CO
2
2
for
for
growth
growth
Thermophilic (42°C)
Thermophilic (42°C)
Microaerophilic
Microaerophilic
Oxidase
Oxidase
-positive
-positive
Virulent factors of
Virulent factors of
Campylobacter
Campylobacter
spp.
spp.
Fimbriae: adherence, penetration
Fimbriae: adherence, penetration
through mucose layer
through mucose layer
Toxins:
Toxins:
Enterotoxin (similar to cholerae toxin)
Enterotoxin (similar to cholerae toxin)
Cytotoxins
Cytotoxins
Epidemiology
Epidemiology
Reservoir: animals
Reservoir: animals
Way of infection: fecal – oral root,
Way of infection: fecal – oral root,
direct contact, birth
direct contact, birth
Minmal amount for infection: 1000
Minmal amount for infection: 1000
bacieria
bacieria
Season infections, most often in winter
Season infections, most often in winter
Most often infected: children
Most often infected: children
Pathogenicity
Pathogenicity
of
of
Campylobacter
Campylobacter
spp.
spp.
C. jejuni
C. jejuni
:
:
A
A
cute gastroenteritis (enterotoxin and
cute gastroenteritis (enterotoxin and
cytotoxin)
cytotoxin)
:
:
Watery and mucos diarhea with blood
Watery and mucos diarhea with blood
Abdominal pain
Abdominal pain
Mild fever
Mild fever
C. fetus
C. fetus
:
:
gastroenteritis,
gastroenteritis,
Treatment of campylobacteriosis:
Treatment of campylobacteriosis:
Macrolides
Macrolides
and tetracyclines
and tetracyclines
Resistance to fluoroquinolones
Resistance to fluoroquinolones
Bacterial examination
Bacterial examination
Speciment: stools, vomited matter, swabs
Speciment: stools, vomited matter, swabs
from rectum
from rectum
Slide (Gram staining) Gram negative
Slide (Gram staining) Gram negative
curved rods, polimorphic
curved rods, polimorphic
Culture (chocolate agar, Brucella agar with
Culture (chocolate agar, Brucella agar with
5%blood, Campylobacter medium, 42
5%blood, Campylobacter medium, 42
°
°
C,
C,
10% CO
10% CO
2
2
)
)
Biochemic reactions
Biochemic reactions
PCR
PCR
Helicobacter pylori
Helicobacter pylori
(formerly
(formerly
Campylobacter
Campylobacter
pylori
pylori
)
)
Gram
Gram
-negative
-negative
Curved
Curved
rods
rods
Motile
Motile
Microaerophilic
Microaerophilic
Urease-producting
Urease-producting
Pathogenicity: chronic
Pathogenicity: chronic
gastritis, gastric and
gastritis, gastric and
duodenal ulcer, gastric
duodenal ulcer, gastric
carcinoma
carcinoma
Determination of
Determination of
pathogenicity:
pathogenicity:
cytotoxin Vac A
cytotoxin Vac A
that causes
that causes
vacuolation in gastric cells
vacuolation in gastric cells
and other factors
and other factors
Virulence factors
Virulence factors
Adhesins
Adhesins
Extraordinary movement in gastric mucos (shape
Extraordinary movement in gastric mucos (shape
+ fimbriae)
+ fimbriae)
Activation of inflammatory respons
Activation of inflammatory respons
Cicoidal form: possibility to survive in unfavorable
Cicoidal form: possibility to survive in unfavorable
conditions
conditions
Cytotoxin VacA:
Cytotoxin VacA:
vacuolation in gastric cells
vacuolation in gastric cells
-
-
destruction of mucos producing cells – expose of
destruction of mucos producing cells – expose of
underlying connective tissue to acid
underlying connective tissue to acid
CagA protein: ulcers
CagA protein: ulcers
Urease – cleave urea producing NH
Urease – cleave urea producing NH
3
3
that
that
neutralizes stomach acid
neutralizes stomach acid
Catalase, superoxide dysmutase - ptoection
Catalase, superoxide dysmutase - ptoection
against superoxide ions (inside phagocytes)
against superoxide ions (inside phagocytes)
Epdemiology
Epdemiology
way of infection: direct contact
way of infection: direct contact
Frequency rate of colonization
Frequency rate of colonization
increases with age , in adults about
increases with age , in adults about
50%
50%
enviromental differentiation is
enviromental differentiation is
observed
observed
Clinical significance
Clinical significance
Acute gastritis (sometimes with
Acute gastritis (sometimes with
diarrhea )
diarrhea )
Gastric ulcer
Gastric ulcer
Duodenal ulcer
Duodenal ulcer
Risk factor for gastric carcinoma
Risk factor for gastric carcinoma
Gastric ulcer
Gastric ulcer
“
“
Gold standard” approach
Gold standard” approach
involves
involves
Endoscopy
Endoscopy
Biopsy – supplies mucosa to be used
Biopsy – supplies mucosa to be used
for Gram stain analysis, histologic
for Gram stain analysis, histologic
examination, culture and rapid tests
examination, culture and rapid tests
(urease production)
(urease production)
Tests of urease activity
Tests of urease activity
Invasive:
Invasive:
Campylobacter-like (CLO) test
Campylobacter-like (CLO) test
Non-invasive:
Non-invasive:
Breath test, Urine test
Breath test, Urine test
Pathogenicity and
Pathogenicity and
diagnostics of anaerobic
diagnostics of anaerobic
bacteria.
bacteria.
Gram-positive and
Gram-positive and
Gram-negative anaerobic
Gram-negative anaerobic
bacteria
bacteria
Anaerobic bacteria genera of
Anaerobic bacteria genera of
medical interest
medical interest
►
Spore-forming rods:
Spore-forming rods:
Clostridium
Clostridium
[Gram(+)]
[Gram(+)]
►
Non-spore-forming rods:
Non-spore-forming rods:
Actinomyces,
Actinomyces,
Nocardia, Bifidobacterium,
Nocardia, Bifidobacterium,
Eubacterium, Lactobacillus,
Eubacterium, Lactobacillus,
Propionibacterium
Propionibacterium
[Gram(+)];
[Gram(+)];
Bacteroides, Fusobacterium, Prevotella,
Bacteroides, Fusobacterium, Prevotella,
Porphyromonas, Gardnerella
Porphyromonas, Gardnerella
[Gram(-)]
[Gram(-)]
►
Cocci:
Cocci:
Peptococcus, Peptostreptococcus,
Peptococcus, Peptostreptococcus,
Streptococcus
Streptococcus
[Gram(+)];
[Gram(+)];
Veillonella
Veillonella
[Gram (-)]
[Gram (-)]
Clostridium
Clostridium
spp.
spp.
Anaerobic spore-forming bacteria
Anaerobic spore-forming bacteria
!!!
!!!
Severe pathogens:
Severe pathogens:
►
C. perfringens
C. perfringens
►
C. difficile
C. difficile
►
C. tetani
C. tetani
►
C.
C.
b
b
otulinu
otulinu
m
m
Others (
Others (
C. novyi, C. histolyticum
C. novyi, C. histolyticum
,
,
C. septicum, C. sporogenes
C. septicum, C. sporogenes
, C. bifermentans
, C. bifermentans
)
)
General characteristics of
General characteristics of
Clostridium
Clostridium
spp.
spp.
►
Gram
Gram
-positive
-positive
►
Long (2-5
Long (2-5
μ
μ
m)
m)
rods
rods
►
A
A
naerobic
naerobic
►
Most species motile
Most species motile
►
Spores located in
Spores located in
different parts of rods -
different parts of rods -
change the shapes of
change the shapes of
bacteria
bacteria
►
Reside in soil and in
Reside in soil and in
digestive tract
digestive tract
Clostridium
Clostridium
perfringens
perfringens
►
The only one non-
The only one non-
motile species
motile species
among
among
Clostridium
Clostridium
spp.
spp.
►
In tissue forms
In tissue forms
capsules
capsules
►
Spores located
Spores located
subterminally
subterminally
Virulence factors of
Virulence factors of
C.
C.
perfringens
perfringens
(division into
(division into
A
A
, B, C,
, B, C,
D,
D,
E serotypes)
E serotypes)
►
Exotoxins:
Exotoxins:
(12 kinds):
(12 kinds):
ά
ά
-toxin
-toxin
– the most important one – lecithinase,
– the most important one – lecithinase,
degradate lecithin in cell membrane causing lysis of
degradate lecithin in cell membrane causing lysis of
endothelial cells, erythrocytes, leucocytes,
endothelial cells, erythrocytes, leucocytes,
(all strains)
(all strains)
Other exotoxins – hemolytic, cytotoxic, necrotic effect
Other exotoxins – hemolytic, cytotoxic, necrotic effect
A – E groups based on exotoxin spectrum
A – E groups based on exotoxin spectrum
►
Enterotoxin
Enterotoxin
: disrupts ion and glucose transport
: disrupts ion and glucose transport
in epithelial cells of small intestine, leading to
in epithelial cells of small intestine, leading to
lose of fluid and intracellular proteins
lose of fluid and intracellular proteins
►
Degradative enzymes:
Degradative enzymes:
DNA-ses,
DNA-ses,
hyaluronidases, collagenase, proteases – role in
hyaluronidases, collagenase, proteases – role in
spread of infection
spread of infection
Pathogenicity of
Pathogenicity of
C.
C.
perfringens
perfringens
►
Gas gangrene
Gas gangrene
►
Food poisoning
Food poisoning
►
Enteritis necroticans
Enteritis necroticans
Gas gangrene
Gas gangrene
Bacteriological
Bacteriological
examination
examination
►
Material: sme
Material: sme
a
a
rs from wounds, f
rs from wounds, f
a
a
eces
eces
,
,
tissue, vomited matter
tissue, vomited matter
►
Media: blood agar or egg yolk agar with
Media: blood agar or egg yolk agar with
neomycin
neomycin
►
Anaerobic conditions
Anaerobic conditions
►
View: in microscope and by the naked eye
View: in microscope and by the naked eye
►
Colonies: large, smooth, convex,
Colonies: large, smooth, convex,
hemolysis
hemolysis
on blood agar
on blood agar
►
Biochemical tests
Biochemical tests
Antibiotic therapy of
Antibiotic therapy of
C. perfringens
C. perfringens
infection
infection
ENTERITIS
ENTERITIS
NECROTICANS
NECROTICANS
►
Penicillin
Penicillin
►
Chloramphenicol
Chloramphenicol
GAS GANGRENE
GAS GANGRENE
►
Surgery(!)
Surgery(!)
►
Penicillin G
Penicillin G
►
Chloramphenicol
Chloramphenicol
►
Metronidazole
Metronidazole
►
Imipenem
Imipenem
Clostridium
Clostridium
tetani
tetani
►
Causative agent of
Causative agent of
tetanus
tetanus
(lockjaw)
(lockjaw)
►
Spores formed at
Spores formed at
the end of bacterial
the end of bacterial
cells (
cells (
drumstick
drumstick
appearance
appearance
)
)
►
Toxins:
Toxins:
tetanospasmin
tetanospasmin
and
and
tetanolysin
tetanolysin
Toxins of
Toxins of
C. tetani
C. tetani
►
Tetanospasmin
Tetanospasmin
(spasmogenic neurotoxin) –
(spasmogenic neurotoxin) –
two subunits: B
two subunits: B
–
–
binding and cell
binding and cell
penetration, A
penetration, A
–
–
blocks
blocks
the
the
inhibitory
inhibitory
neurotransmitter
neurotransmitter
release at central and
release at central and
peripheral synapses
peripheral synapses
resulting in
resulting in
accumulation of acetylocholine and
accumulation of acetylocholine and
unrestrained excitation of the motor
unrestrained excitation of the motor
neurons.
neurons.
►
Tetanolysin
Tetanolysin
-
-
hemolytic toxin
hemolytic toxin
Toxins penetrating into bloodstream are
Toxins penetrating into bloodstream are
responsible for clinical
responsible for clinical
symptoms
symptoms
Ttetanus
Ttetanus
Therapy of
Therapy of
C. tetani
C. tetani
infection
infection
►
Specific antiserum neutralizing
Specific antiserum neutralizing
bacterial toxins
bacterial toxins
►
Surgical removal of infected tissue
Surgical removal of infected tissue
►
Antibiotics
Antibiotics
– penicillin
– penicillin
Prevention
Prevention
vaccines consisting of bacterial
vaccines consisting of bacterial
anatoxins
anatoxins
Clostridiu
Clostridiu
m
m
botulinum
botulinum
►
Casuative agent
Casuative agent
of botulism
of botulism
►
Spores situated
Spores situated
subterminally
subterminally
Virulence factors of
Virulence factors of
C.
C.
botulinum
botulinum
►
Neurotoxins (A-F) called botulin
Neurotoxins (A-F) called botulin
(exotoxin)
(exotoxin)
: in peripheral cholinergic
: in peripheral cholinergic
synapses
synapses
blocks acetylocholine
blocks acetylocholine
relase
relase
causing flaccid paralysis
causing flaccid paralysis
►
Toxins active after bacterial lysis
Toxins active after bacterial lysis
►
Botulin is heat-sensitive (boiling for
Botulin is heat-sensitive (boiling for
20 min)
20 min)
Infant botulism
Infant botulism
Treatment
Treatment
of botulism
of botulism
►
Penicillin
Penicillin
►
Antitoxin
Antitoxin
Clostridium difficile
Clostridium difficile
►
Casuative
Casuative
agent of
agent of
antibiotic-
antibiotic-
associated
associated
diarrheas
diarrheas
( AAD)
( AAD)
►
Spores situated
Spores situated
terminally
terminally
Virulent factors of
Virulent factors of
C.
C.
difficile
difficile
►
Cytotoxins
Cytotoxins
A: enterotoxin, stimulation of
A: enterotoxin, stimulation of
inflammatory response
inflammatory response
B: cytotoxin - damage epithelial cells
B: cytotoxin - damage epithelial cells
Clostridium difficile
Clostridium difficile
–
–
pathogenicity
pathogenicity
►
After antibiotics (especially clindamycin,
After antibiotics (especially clindamycin,
ampicillin
ampicillin
,
,
and cephalosporines)
and cephalosporines)
:
:
Antibiotic-associated diarrhea (toxin A
Antibiotic-associated diarrhea (toxin A
)
)
Antibiotic-associated colitis
Antibiotic-associated colitis
Antibiotic-associated pseudomembranous
Antibiotic-associated pseudomembranous
colitis
colitis
TREATMENT
TREATMENT
►
vancomycin, teicoplanin
vancomycin, teicoplanin
Bacteroides fragilis
Bacteroides fragilis
group
group
►
Gram
Gram
-negative
-negative
rods
rods
, h
, h
igh
igh
degree of pleomorphism
degree of pleomorphism
►
Non-motile
Non-motile
►
Non-sporing
Non-sporing
►
C
C
apsuled
apsuled
►
Anaerobi
Anaerobi
c
c
►
Resistant to:
Resistant to:
bile,
bile,
kanamycin, vancomycin,
kanamycin, vancomycin,
colistin
colistin
►
Hydrolyse
Hydrolyse
s
s
esculine
esculine
►
Some produce dark
Some produce dark
pigments (black colonies)
pigments (black colonies)