Gram Positive Bacilli

Gram Positive Bacilli can be divided into:

Spore Formers Non-Spore Formers
Bacillus Clostridium Corynebacteria
Aerobes Anaerobes (Clos-) Erysipelothrix rhusiopathiae – H2S 
Catalase  Catalase (-) Listeria monocytogenes
No gas production Gas producers Lactobacillus

Bacillus spp.
Bacillus anthracis (anthrax bacillus)  On Gram Stain: Bamboo fishing rod appearance
 Penicillin Susceptible – w/ ‘String of Pearl’ appearance on
Bioterrorism Agent! MHA with 10u Penicillin
Causative agent of:  Lecithinase  = opaque zone around colonies on egg yolk
Cutaneous Anthrax - Most common
Pulmonary Anthrax (“Woolsorter’s agar
Disease”)  Gelatin medium: “Inverted Pine Tree” appearance Ascoli Test
Gastrointestinal Anthrax – test to detect anthrax antigens Capsule: D-glutamic acid
On BAP: Non-hemolytic, w/ medusa-head or
beaten egg white appearance

Bacillus cereus (Fried Rice Bacillus) Bacillus subtilis (Hay Bacillus)

 Cholera-like toxin causes food poisoning  Laboratory Contaminant
 Also Lecithinase   Produces Bacitracin
 Frosted glass appearance on BAP  Ground glass appearance on BAP

Capsule Hemolysis Motility Penicillin

B. anthracis Positive Negative Negative Susceptible
B. cereus Negative Positive Positive Resistant

Geobacillus stearothermophilus – indicator for autoclave (steam sterilization)

Bacillus subtilis var. niger (now B. atropheus) – indicator for dry heat sterilization
Clostridium
1. Clostridium botulinum (Canned food bacillus)
 Causative agent of botulism (flaccid paralysis)
 Virulence factor: botulinum toxin – the most potent toxin
 Lipase 
2. Clostridium tetani (Tack Head Bacillus)
 Causative agent of tetanus (spasmic paralysis)
o Associated terms: opisthotonos, devil’s grin
o Virulence factors: tetanolysin and tetanospasmin
 Motile, fine spreading film on culture
 Terminal Spores: “Drumstick/tennis racquet appearance”
 Biochemical:  Gelatinase, Indole; Negative for Lecithinase and Lipase
3. Clostridium perfringens
 Former name: Clostridium welchii
 Causative agent of gas gangrene (myonecrosis)
 Capsulated, nonmotile, subterminal spores
 Virulence factors: alpha toxin = lecithinase; beta toxin, enterotoxin – food poisoning
 Tests:
o Ferments Lactose, Sucrose; aerogenic
o Litmus Milk: acidified with stormy clot formation
o On SBAP: double zone hemolysis
o Reverse CAMP Positive with Group B Strep (S. agalactiae)
o Nagler Reaction: to detect lecithinase activity
  result = opacity on medium with egg yolk / human serum
 Specific result = neutralized with anti-alpha toxin
4. Clostridium difficile
 Causative agent of pseudomembranous ulcerative colitis (antibiotic associated diarrhea)
 Media: CCFA (Cycloserine Cefoxitin Fructose Agar); ferments Fructose
 = ground-glass colonies with yellow halo
 BAP: “Horse stable” odor; non hemolytic; fluorescent under UV light

Motility Lecithinase Lipase Lactose Glucose

C. perfringens Nonmotile  Negative 

C. botulinum 
C. tetani  Negative
Negative
Negative Negative
C. difficile 
Corynebacterium spp. – Catalase , Oxidase 
 Non motile; Highly Pleomorphic:
 On Gram stain: Chinese Characters, Palisading/Picket fence, club shaped

Corynebacterium diptheriae aka Klebs-Loeffler bacillus

 Causative agent of diphtheria
 Pseudomembrane formation and bull’s neck
 Urease Negative; Nitrate 
 Metachromatic granules (Babe Ernst granules) Tests:
o Visualized with Loeffler’s Methylene Blue In Vitro Toxigenicity: Elek Test
 Enhanced by Pai Agar and Loeffler’s Serum Slant In Vivo Toxigenicity: Animal Inoculation
Susceptibility: Schick’s Test

Colonial types of C. diptheriae (accdg. Delost)

Gravis 1 to 2 mm colonies on blood agar; largest colonial type

Mitis Fried egg appearance on BAP; bleach-like odor on tellurite medium
Intermedius Small colonies (0.5 mm) on blood agar; black colonies with gray
borders on tellurite medium
*some references mention a fourth: belfanti

Media: Other Corynebacterium spp.

 Tindale’s Medium
 Cystine Tellurite Blood Agar (CTBA)
– preferred medium for isolation and
ID of Corynebacteria
o  - Brown halo around colonies:
C. diptheriae, C. ulcerans, C.
pseudotuberculosis
 C. urealyticum
 urinary pathogen
 BAP: Pinpoint, white, smooth, non-hemolytic
 Rapid Urease Producer
 C. jeikium
Infects immunocompromised patients
Negative for Urease and Nitrate
 C. pseudodipthericum (Hoffman Bacillus)
 Does not exhibit pleomorphism

Listeria monocytogenes
 Listeriosis: Food poisoning, Meningitis
 Virulence Factor: Listeriolysin O
 Catalase , H2S Negative, Motile
 Beta hemolytic, CAMP + (Note: Uses Rhodococcus equi)
 On wet prep (Hanging drop): Tumbling Motility
 On semisolid media (SIM) at RT (25 C): Inverted Christmas Tree / Umbrella
 Selective Media: McBride Medium
 Virulence test: Ocular Test of Anton
 Cold Enrichment test: Nutrient Broth, 4 degrees Celsius
o Also positive: Yersinia enterocolitica

May resemble Group B Streptococci To differentiate from Corynebacteria

Group B Strep L. monocytogenes LM C
Motility Negative  Motility  Negative
Catalase Negative  Salicin  Negative
CAMP “arrowhead” “block” Fermentation

Erysipelothrix rhusiopathiae Lactobacillus acidophilus

 Causative agent of Erysipeloid
o Erysipelas: Streptococcus pyogenes
 Catalase Negative, H2S , Non-motile
 On semisolid media: Test Tube Brush appearance
 Associated with: livestock (veterinary pathogen)
 Normal flora of mouth, GIT, and vaginal canal
 Vaginal canal: Doderlain bacillus
 GI Tract: Boas-Oppler bacillus
 Negative for: Catalase, Motility, H2S, Bile Esculin
 Media: Tomato Juice Agar