Pathogenesis and Clinical Features

• Three types of botulism are recognised: foodborne botulism, infant or

infectious botulism and wound botulism. Only in the first type is food
invariably involved.
• Foodborne botulism is an example of bacterial food poisoning.
• Unlike enterotoxins, which act locally in the gut, they affect primarily
the cholinergic nerves of the peripheral nervous system.
 Pathogenesis and Clinical Features (Cont.)

• Experiments in animals have shown that toxin ingested with food and
surviving inactivation is absorbed in the upper part of the small
intestine and reaches the bloodstream via the lymphatics.
• It binds to the nerve ending at the nerve–muscle junction, blocking
release of the acetylcholine responsible for transmission of stimuli,
thus producing a flaccid paralysis.
• Survival is therefore critically dependent on early diagnosis and
treatment, principally by alkaline stomach washing to remove any
remaining toxic food, intravenous administration of specific or
polyvalent anti-toxins to neutralize circulating toxin, and mechanical
respiratory support where necessary.
 Isolation and Identification

• In view of the metabolic diversity within the species selective media

are of limited use in the isolation of C. botulinum and identification is
based on the ability of typical colonies to produce toxin in culture.
• C. botulinum will often constitute only a small proportion of the total
microflora so enrichment or pre-incubation is necessary to improve the
chances of isolation. Sometimes enrichment cultures are heated prior
to incubation to eliminate non-sporeforming anaerobes
 Isolation and Identification(Cont.)
• After enrichment in a medium such as cooked meat broth at 30 1C for
7 days, the culture is streaked on to fresh horse-blood or egg yolk agar
and incubated anaerobically for 3 days.
• Characteristic smooth colonies, 2–3mm in diameter with an irregular
edge and showing lipolytic activity on egg-yolk agar (type G
excepted) are transferred into a broth medium to check for toxin
production.
• A technique has been described that simplifies this procedure by
incorporating antitoxin into the agar medium so that toxin-producing
colonies are surrounded by a zone of toxin–antitoxin precipitate.
 Isolation and Identification(Cont.)
• Despite the development of a range of in vitro immunoassay
procedures for toxin, the mouse neutralization test remains the most
sensitive (a typical lethal dose of toxin for a mouse is a few
picograms).
• However, the distressing nature of the test guarantees its eventual
replacement as soon as immunoassay amplification systems have been
sufficiently improved.
• The presence of toxin is confirmed by protection of mice with
polyvalent antitoxin and the toxin type can be identified using
monovalent antisera.