Clin Bacte Mod 3 (G + Spore F: Bacillus and Clostridium)

Bacillus OXYGEN

+ (STRICT AEROBE)

Clostridium OXYGEN

- (STRICT ANAEROBE)

Bacillus CATALASE (15% H202)

+

Clostridium CATALASE (15% H202)

-

Bacillus anthracis is also called?

Bacillus anthracis

Category A Bioterrorism agent

Bacillus anthracis

Bacillus anthracis motility and hemolysis

non-motile and gamma hemolytic

Location of B. anthracis spores

central

Media used to isolate B. anthracis

5% SBA

B. anthracis selective media

PLET — Polymyxin Lysozyme EDTA Thallous acetate

PLET — Polymyxin Lysozyme EDTA Thallous acetate pattern

shows “string-like” pattern because of its susceptibility to penicillin

Appearance of B. Anthracis under the microscope

forms DISJOINTED BAMBOO FISHING ROD appearance because of the squared ends

Appearance of B. Anthracis in MHA media

String of Pearl, — due to susceptibility to penicillin

Demo of capsule of B. anthracis

M’ Fadyean using polychrome methylene blue

Demo SPORES of B. anthracis

Schaeffer and Fulton: spores appear green (due to Malachite Green)

B. anthracis in BAP (5% SBA)

• nonhemolytic

• colonies with swirling projections, “MEDUSA HEAD” or “LION HEAD” or “COMET TAIL” appearance

• colonies with “cut glass” appearance

B. anthracis in TENACIOUS COLONIES

shows BEATEN EGG WHITE consistency

B. anthracis in GELATIN Media

forms so-called “INVERTED FIR TREE/INVERTED PINE TREE” pattern

Virulence Factors of B. anthracis

• Polypeptide D-glutamic acid capsule

• production of EXOTOXIN with 3 components

EXOTOXIN with 3 components

• Edema Factor (EF)

• Lethal Factor (LF)

• Protective Antigen

most common type and least severe of Anthrax

CUTANEOUS ANTHRAX

MOT: Contact with infectious spores while handling wool (sheep)

CUTANEOUS ANTHRAX

develop “BLACK ESCHAR”

CUTANEOUS ANTHRAX

PULMONARY ANTHRAX is also called

• “Woolsorter’s Disease”

• “Ragpicker’s Disease”

• “Hide Porter’s Disease”

MOT: Inhalation of spores while handling wool

PULMONARY ANTHRAX

INTESTINAL ANTHRAX

least common type and most severe of anthrax

MOT: Ingestion of spores (improperly cooked infected meet)

INTESTINAL ANTHRAX

INJECTIONAL ANTHRAX

rare form of anthrax; may develop among addicts

MOT: Use of contaminated drugs of abuse (ex. HEROIN)

INJECTIONAL ANTHRAX

LECITHINASE TEST media

- egg yolk agar

(+) result for LECITHINASE TEST

opaque zone around the colonies (MOTHER OF PEARL COLOR)

also called (Fried Rice Bacillus)

Bacillus cereus

Bacillus cereus Causes food poisoning

— consumption of contaminated rice meals

Emetic

nausea and vomiting 1-5 hrs (incubation period)

Diarrheal

abdominal pain and diarrhea; 8-16 hrs (incubation period)

Virulence Factors of Emetic

Cereulide

Virulence Factors of Diarrheal

• Nhe (Non-hemolytic enterotoxin),

• cytK (cytotoxin K) or Hemolysin IV

• HBL

Best specimen for testing B. cereus

suspected food

Culture Media of B. cereus

• MEYP - Mannitol Egg Yolk Polymyxin B agar

• PEMBA - Polymyxin Egg Yolk Mannitol Bromthymol blue agar

Which are motile and beta hemolytic on BAP?

• B. cereus

• B. subtillis

Bacillus subtilis (Hay Bacillus) is under what BSL?

BSL1 - common laboratory contaminant

Bacillus subtilis (Hay Bacillus) is resistant to?

penicillin

Bacillus subtilis (Hay Bacillus) selective media

MEYP

What Causes eye infection in heroin addicts?

Bacillus subtilis (Hay Bacillus)

a blood bank contaminant

Bacillus subtilis (Hay Bacillus)

Bacillus subtilis (Hay Bacillus) is a _____ pathogen, a source of

opportunistic, bacitracin

Sterilization

all are removed

Disinfection

only pathogens are removed

Methods for Sterilization:

• gamma rays

• moist heat

• dry heat

Autoclaving biological indicator

B. stearothermophilus

Autoclaving

• KILLS SPORES

• to sterilize with heat

• monitor temperature and efficiency: daily

biological indicator of OVEN

B. subtilis var. niger:

for maintenance

B. subtilis var. niger:

B. anthracis

• Hemolysis in SBA -

• Motility -

• Penicillin susceptibility S (+)

• Lecithinase production +

• Fermentation of salicin -

• Growth in penicillin (10 U/mL) agar -

• Gelatin hydrolysis -

• Growth on phenylethyl alcohol agar -

B. cereus

• Hemolysis in SBA +

• Motility +

• Penicillin susceptibility R (-)

• Lecithinase production +

• Fermentation of salicin +/-

• Growth in penicillin (10 U/mL) agar -

• Gelatin hydrolysis -

• Growth on phenylethyl alcohol agar -

Bacillus thuringiensis

insect pathogen

can be observed using phase contrast microscopy or spore staining

Bacillus thuringiensis

NEUROTOXIC

causes paralysis

NEUROTOXIC bacteria

• Clostridium tetani

• Clostridium botulinum

HISTOTOXIC

affects tissues

HISTOTOXIC bacteria

Clostridium perfringens

ENTERIC

affects GIT

ENTERIC bacteria

Clostridium difficile

Clostridium difficile

Normal flora of the colon

can cause PSEUDOMEMBRANOUS COLITIS or ANTIBIOTIC-ASSOCIATED DIARRHEA

Clostridium difficile

SEUDOMEMBRANOUS COLITIS or ANTIBIOTIC-ASSOCIATED DIARRHEA is due to

to long term antibiotic therapy; CLINDAMYCIN

Virulence factors of C. difficile

Toxin A & B

Toxin A

enterotoxin

Toxin B

cytotoxin

Cytotoxin Detection

EIA

C. difficile Culture

CCFA (Cefoxitin Cyclosporine Fructose Agar)

BAP

CCFA (Cefoxitin Cyclosporine Fructose Agar) pH indicator

neutral red

CCFA (Cefoxitin Cyclosporine Fructose Agar) colonies

HORSE TABLE / BARNYARD ODOR

BAP in C. difficile

fluoresce CHARTREUSE (yellow fluorescence)

SPECIMEN for C. difficile

• Freshly passed, LIQUID stool

• Formed stool / Rectal Swab

Freshly passed, LIQUID stool

for culture and toxin assay (EIA)

Formed stool / Rectal Swab

– detection of CARRIER STATE — normal presence/normal flora

Clostridium botulinum is also called

Home Canned Good Bacillus

What clostridium is not cultured?

C. botulinum

Von ermengen’s Bacillus

Clostridium botulinum

Clostridium tetani is also called

Tennis Racquet, Lollipop, Drumstick Bacillus, Tack Head Bacillus

Iron-loving

Clostridium tetani

What causes Tetanus?

Clostridium tetani

Virulence factor of C.botulinum

Botulinum toxin (BOTOX) - neurotoxin

Botulinum toxin (BOTOX) blocks release of _______- that causes?

acetylcholine, FLACCID paralysis

FLACCID paralysis

muscles are unresponsive

BOTULISM

Fatal type of FOOD POISONING

Infant Botulism:

may develop as a result of SPORE INGESTION via BREAST FEEDING

Infant Botulism symptom

FLOPPY BABY SYNDROME (hypotonia)

What is to detect LIPASE production of C. botulinum?

EYA

(+) of EYA

opaque zone around colonies

Virulence Factor of C.tetani

Tetanospasmin – neurotoxin

Tetanospasmin block release of neurotransmitters causing

• SPASTIC paralysis

• LOCK JAW

SPASTIC paralysis

very limited/little movement

LOCK JAW if prolonged causes

RISUS SARDONICUS

Clinical symptoms of C.tetani

• Tetanic Triad

• Observation of TERMINAL SWOLLEN SPORES

Tetanic Triad includes

• Trismus

• Risus Sardonicus

• Opisthotonos

Trismus

“Lockjaw” - mild → severe spasms of lower jaw

Risus Sardonicus

“Sardiman Grin” - grin caused by facial muscle spasm

Opisthotonos

severe simultaneous spasm of all muscles