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)