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There are reports of a hitherto unknown bovine disease in Germany. The symptoms are, in general, indigestion (constipation alternating with diarrhoea), non-infectious chronic laminitis, engorged veins, oedemas, retracted abdomen, emaciation and apathy. Most cases occur during the peripartal period and often result in unexpected death. In addition, there are findings of delayed growth and wasting in heifers, as well as decreasing milk yield. Clinical and standard laboratory examinations leave the origin undisclosed. Bioassays for Clostridium botulinum, its spores and toxins in animals of affected farms revealed the presence of free botulinum toxin in the contents of the lower sections of the intestine. In two control farms without signs of the disease, the tests remained negative. This seems to support our hypothesis that long-lasting absorption of low quantities of botulinum toxin may interfere with the neurological control of intestinal physiology. The authors propose to name this disease complex 'visceral botulism'.

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A sensitive and specific immunoassay for the simultaneous detection of Clostridium botulinum type C (BoNT/C) and type D neurotoxin was developed. Goat anti-mouse immunoglobulin G was bound to polyethylene disks in a small disposable column used for this assay. The sample was preincubated together with monoclonal antibodies specific for the heavy chain of BoNT/C and D and affinity-purified, biotinylated polyclonal antibodies against these neurotoxins. This complex was captured on the assay disk. Streptavidinpoly-horseradish peroxidase was used as a conjugate, and a precipitating substrate allowed the direct semiquantitative readout of the assay, if necessary. For a more accurate quantitative detection, the substrate can be eluted and measured in a photometer. Depending on the preincubation time, a sensitivity of 1 mouse lethal dose ml ؊1 was achieved in culture supernatants.Clostridium botulinum is a gram-positive, anaerobic, and spore-forming soil bacterium that can be found in the guts of domestic animals (9, 10). The fact that it produces the most toxic metabolite known to humans brought it to the attention of medical microbiologists in the early days of this field (27). Nowadays, seven distinct botulinum neurotoxins (BoNTs) are known. In the order of their discovery, they have been named types A to G. However, the capability to produce BoNTs is not limited to C. botulinum only. Some strains of Clostridium baratii (8) and Clostridium butyricum (3, 21) are toxigenic as well.The prevalence of C. botulinum in the soil might explain where the bacterium enters the food chain (20), leading to the best-known form of the disease in the worst case, food-borne botulism (25). The bacterium multiplies in food or feed under favorable conditions and produces the toxin, which is orally taken up by the host. The typical signs of flaccid paralysis develop, which are caused by the inhibition of acetylcholine release at the synapses (19).In animal husbandry and for wildlife, types C and D botulism are predominant. Throughout the world, millions of waterfowl have reportedly died from botulism caused by BoNT type C (BoNT/C) (24). BoNT/C and D are pathogenic for our domestic animals, with sometimes dramatic losses in the affected farms (6, 15). The losses of cattle reported from Brazil amount to five million animals over the past 10 years (18).However, the disease may present as a toxico infection as well. With the shaker foal syndrome in horses, it was shown that the bacteria colonize the gut and produce the toxin in the host animal (23). Visceral botulism in cattle (5) and equine grass sickness (7) might have a toxico infectious botulinum etiology as well.The recent concerns for the use of botulinum neurotoxins by bioterrorists (2) again highlighted the fact that only a limited number of tools are available to detect BoNTs. The mouse bioassay, still the most common method, needs to be replaced for obvious reasons. Recently, alternative in vitro tests (13, 17) have become commercially available. However, these assays are limited t...

the results show a remarkable incidence of infant botulism without any known previous medical history, partly hidden as sudden infant death. We propose to systematically search for botulism in connection with sudden infant death.