MIC 251

ENDOSPORE-FORMING
GRAM POSITIVE RODS
Lecture 22

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Learning outcomes
◦ Identify the general properties and diseases caused
by medical important species in this groups.
◦ Organisms focused on
◦ Bacillus sp.
◦ Clostridium sp.

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Endospore-forming Gram Positive rods
◦ Contains 3 classes
◦ Mollicutes
◦ Bacilli (10 orders ; 34 families)
◦ Clostridia

◦ Mollicutes
◦ Do not have a cell wall
◦ E.g. Mycoplasma (See lecture 20)

◦ Bacilli and Clostridia

◦ Contains cell walls
◦ Ubiquitous in the environment
◦ Form endospores and can survive for many years
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 Bacillus sp.
◦ Contains 2 orders
◦ Bacillales and Lactobacillales (17 families ; over 70 genera)

◦ Endospore formation were also important in the phonetic classification of the first edition of Bergey’s Manual
◦ Order Bacillales
◦ Divided into several families with important genera
◦ Family Baciallaceae
◦ Genus Bacillus
◦ Family Caryophanaceae
◦ Genus Caryophanon
◦ Family Listeraceae
◦ Genus Listeria
◦ Family Staphylococcaceae
◦ Genus Staphylococcus
◦ Family Thermoactinomycetaceae
◦ genus Thermoactinomyces
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 Endospore-forming Gram positive rods
Peptidoglycan Structure (diamino-pimelic acid DAP) – cell wall surrounding cytoplasmic membrane
◦ Varies considerably among different gram positive groups
◦ Most gram negatives have a peptidoglycan structure in which meso-DAP in position 3 is directly cross-linked
through its free amino group to the free carboxyl group of the terminal D-Ala of an adjacent peptide chain 
also present in many gram positives.

◦ In some gram positives lysine is

substituted for DAP in position 3 and
the peptide subunits are cross-linked by
interpeptide bridges

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Bacillus
◦ Characteristics
◦ Endospore forming
◦ Chemoheterotrophic rods
◦ Usually motile with peritrichous flagella
◦ Gram positive rods (1 x 3-4µm) and often occurs in chains
◦ Spores located in centre of non-motile bacilli
◦ Aerobic or facultative aerobic
◦ Catalase positive
◦ Most members occur in soil, water, vegetation, air

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 Bacillus
◦ Important species of Bacillus
◦ Bacillus subtilis
◦ Important experimental organism – genome sequenced
◦ Families of genes expanded by gene duplication
◦ Various species produces antibiotics
◦ Bacitracin, gramicidin, polymyxin

◦ Bacillus cereus
◦ Food poisoning
◦ Bacillus anthracis
◦ Antrax
◦ Bacillus thuringiensis and Bacillus sphaericus
◦ Used as pesticide
◦ Solid protein crystal that contains toxin
◦ Parasporal body

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Bacillus anthracis
◦ Anthrax
◦ Anthrax usually disease of herbivores
◦ Humans become infected through accidental contact with infected animals or products

◦ 4 forms of Anthrax
◦ Cutaneous
◦ Gastrointestinal
◦ Inhalation
◦ Injection
◦ Spores germinate at site of entry
◦ Growth result in gelatinous edema and congestion
◦ Virulence is dependant on capsule production
◦ Agent of bioterrorism (2001)

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Cutaneous anthrax
◦ Most common
◦ Causes localized inflammation
◦ Black necrotizing lesion is formed
◦ Eschar
◦ Is a slough or piece of dead tissue that casts off the surface of
the lesion

◦ 1-7 days

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Inhalation anthrax
◦ Rare, but highly fatal
◦ Also known as “woolsorters disease”
◦ Inhalation of spores
◦ Presents flu-like symptoms and chest discomfort
◦ Symptoms 1 week after exposure

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Gastrointestinal anthrax
◦ Also rare, but highly fatal
◦ Ingestion of spores
◦ Symptoms present as fever, swelling of neck, painful swallowing, hoarseness, nausea, (bloody) vomiting,
diarrhoea, red eyes, swelling of abdomen
◦ Symptoms 1-7 days

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Injected anthrax
◦ Emerging form of anthrax amongst drug users
◦ Cases have emerged in Northern Europe
◦ Symptoms similar to cutaneous anthrax, however infection may initiate deeper in tissue
◦ Injection can travel faster in system and may be unrecognizable

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Bacillus cereus
◦ Associated with food poisoning
◦ 2 distinct forms
◦ Emetic (vomiting) and diarrhoea
◦ Also causes eye infections

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Clostridium
◦ Contains 3 orders and 11 families

◦ Genus Clostridium
◦ Includes obligate anaerobic, endospore formers
◦ Large rods
◦ Motile with peritrichous flagella
◦ Spores usually wider than the rod
◦ Terminally, sub-terminally, centrally
◦ Natural habitat is soil or intestinal tract of humans and animals
◦ Over 100 species in distinct phylogenetic clusters
◦ Might be divided into several genera
◦ Responsible for many cases of food spoilage
◦ Products of fermentation
◦ Hydrogen sulphide, ammonia, amines
◦ Responsible for he unpleasant odours of V.Jooste
putrefaction
Clostridium
Important spp. of Clostridia
◦ Clostridium botulinum
◦ Food spoilage (especially canned food)
◦ botulinum
◦ Clostridium tetanus
◦ tetanus
◦ Clostridium perfringens
◦ Gas gangrene
◦ Clostridium difficile
◦ Pseudomembraneous colitis

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Clostridium botulinum

◦ Worldwide distribution
◦ Distinguished by antigenic toxin
◦ Neurotoxin
◦ Toxin is heat stable
◦ Intoxication resulting from ingestion of food
◦ Botulism → flaccid paralysis → Inability to swallow → Respiratory paralysis, cardiac
arrest → death
◦ Patient remains conscious until death
◦ Mortality rate is high
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Clostridium tetani
◦ World wide distribution
◦ Soil and animal feces
◦ Distinguishes by specific flagellar antigens
◦ All produce the same type of neurotoxin
◦ tetanospasmin
◦ Disease is entirely a toxaemia (blood poisoning)
◦ Tetanus → spastic paralysis
◦ Muscular spasms → Patient is fully conscious until death
◦ Mortality rate in generalized tetanus is very high

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 Clostridium perfringens
◦ Many different toxin-producing Clostridia cause invasive infection
◦ Most common is C. perfringens
◦ Myonecrosis and gas gangrene

◦ Also produces an enterotoxin that causes food poisoning

◦ Lecithinase (alpha toxin), Dnase, Hyaluronidase, Collagenase

◦ Ferment carbohydrates and produce gas

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Clostridium difficile
◦ Pseudomembrane colitis
◦ Diagnosis is made when toxins are found in both stool and endoscopic observation of
pseudomembrane / microabscesses of individuals that have diarrhoea and have been administered
antibiotics.

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Laboratory diagnosis

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Willis and Hobbs (Lecithinase) plate

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 END OF LECTURE 22
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