The Case of Botulinum Toxin in Milk: Experimental Data

Toxinology Group, Spiez Laboratory, 3700 Spiez, Switzerland.
Applied and Environmental Microbiology (Impact Factor: 3.67). 04/2010; 76(10):3293-300. DOI: 10.1128/AEM.02937-09
Source: PubMed


Botulinum neurotoxin (BoNT) is the most toxic substance known to man and the causative agent of botulism. Due to its high toxicity and the availability of the producing organism Clostridium botulinum, BoNT is regarded as a potential biological warfare agent. Because of the mild pasteurization process, as well as rapid product distribution and consumption, the milk supply chain has long been considered a potential target of a bioterrorist attack. Since, to our knowledge, no empirical data on the inactivation of BoNT in milk during pasteurization are available at this time, we investigated the activities of BoNT type A (BoNT/A) and BoNT/B, as well as their respective complexes, during a laboratory-scale pasteurization process. When we monitored milk alkaline phosphatase activity, which is an industry-accepted parameter of successfully completed pasteurization, our method proved comparable to the industrial process. After heating raw milk spiked with a set amount of BoNT/A or BoNT/B or one of their respective complexes, the structural integrity of the toxin was determined by enzyme-linked immunosorbent assay (ELISA) and its functional activity by mouse bioassay. We demonstrated that standard pasteurization at 72 degrees C for 15 s inactivates at least 99.99% of BoNT/A and BoNT/B and at least 99.5% of their respective complexes. Our results suggest that if BoNTs or their complexes were deliberately released into the milk supply chain, standard pasteurization conditions would reduce their activity much more dramatically than originally anticipated and thus lower the threat level of the widely discussed "BoNT in milk" scenario.

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    • "Nevertheless, we expect that contamination of milk in dairy plants is less likely than in farms due to restricted access to the dairy plants. Relevant characteristics of milk, the kind of biological agents or toxins, individual dispositions like the age of people [34] [35] [36] [37] and internal processes like pasteurization [38] [39] [40] [41] [42] [43], which may influence the vulnerability of the consumers to contamination, but also the spread of contamination [32], are not considered in this paper. "
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    ABSTRACT: Dairy farms were identified, which can be included in a contingency plan set up to prevent or mitigate the consequences of deliberate contamination of a food supply chain. The deliberate introduction of a contamination into the supply chain of milk was simulated in a scenario where milk producers serve as the entry sources and consumers of milk represent the target to be affected by the contamination. It is shown that the entry sources have an impact on the damage caused, i.e. in terms of the number of consumers reached. A contingency plan is provided that contains a list of entry sources ranked according to their impact on the damage to consumers. To generate this list, a computer program was developed that simulates the impact of the contaminations on consumers via the trade of contaminated milk. Possible variations in the trade links between milk producers, dairies and consumers as well as between dairies are considered. It is investigated how these trade links alter the generated list of entry sources. The results indicate that, regardless of the actual milk trade flow, control measures should be introduced on 39% of the milk producers in order to minimize the damage. The identification of suitable entry sources may help risk managers to focus on these farms in a contingency plan that improves the sensitivity of control activities related to deliberate contamination. & 2014 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY-NC-ND license (
    Omega 12/2014; 53. DOI:10.1016/ · 4.38 Impact Factor
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    • "These include neurotoxigenic clostridia (C. botulinum and C. tetani) (Weingart et al., 2010), clostridia involved in gas gangrene and necrotizing infections (C. perfringens, C. sordellii and C. septicum) (Hatheway, 1990; Aldape et al., 2006), and the enteropathogenic C. difficile (Twine et al., 2009). "
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    ABSTRACT: The genus Clostridium is made up of species that cause disease to both human beings and animals, with zoonotic species/strains playing critical roles in disease dynamics. Clostridial organisms posses pathogenic, therapeutic and industrial uses. Pathogenic clostridia cause lethal or life threatening infections that must be treated, while the industrial clostridia produce bio-fuels, to serve as alternative energy sources in the face of very high global prices of conventional fuels. Genetic engineering is currently developing novel strains by replacing or altering the gene configuration of pathogenic strains (gene knockout/down) to convert them to solvenogenic and non pathogenic strains for industrial uses. Therapeutic clostridia serve as vehicles for treatment of diseases, especially solid tumors. Because of the global problem of antibiotic resistance, which is a survival strategy by microbial pathogens, some of the well known chemotherapeutic protocols have failed and there is the need to evolve new treatment approaches, using novel agents that have superior mechanisms of action, compared to conventional antibiotics that are becoming inconsequential because of resistance. It is believed that natural products may be effective alternatives to resolving this puzzle, since they are cheaper, readily available and possibly effective against clostridial species. In this review article, the authors discussed some of the life threatening and solvenogenic clostridia and listed some natural products that may possibly be employed as drug targets for ameliorating the problem of resistance in the future, if their active principles are thoroughly researched. It is concluded that medical and veterinary research should be re-jigged to evolve active principles from natural products that are not so easily surmounted by the surge in drug resistant strains.
    06/2014; 7(2):81-94. DOI:10.12816/0008220
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    • "It has been discussed in the relevant literature [39], whether BoNT/A might be used in case of a bioterror attack to contaminate the food supply, i.e., the milk production. Accordingly, it seemed pivotal to test the bioanalytical system with toxins in raw milk as food matrix. "
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    ABSTRACT: Prevalent incidents support the notion that toxins, produced by bacteria, fungi, plants or animals are increasingly responsible for food poisoning or intoxication. Owing to their high toxicity some toxins are also regarded as potential biological warfare agents. Accordingly, control, detection and neutralization of toxic substances are a considerable economic burden to food safety, health care and military biodefense. The present contribution describes a new versatile instrument and related procedures for array-based simultaneous detection of bacterial and plant toxins using a bioanalytical platform which combines the specificity of covalently immobilized capture probes with a dedicated instrumentation and immuno-based microarray analytics. The bioanalytical platform consists of a microstructured polymer slide serving both as support of printed arrays and as incubation chamber. The platform further includes an easy-to-operate instrument for simultaneous slide processing at selectable assay temperature. Cy5 coupled streptavidin is used as unifying fluorescent tracer. Fluorescence image analysis and signal quantitation allow determination of the toxin's identity and concentration. The system's performance has been investigated by immunological detection of Botulinum Neurotoxin type A (BoNT/A), Staphylococcal enterotoxin B (SEB), and the plant toxin ricin. Toxins were detectable at levels as low as 0.5-1 ng · mL(-1) in buffer or in raw milk.
    Sensors 12/2012; 12(2):2324-39. DOI:10.3390/s120202324 · 2.25 Impact Factor
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