Construction and validation of safe Clostridium botulinum Group II surrogate strain producing inactive botulinum neurotoxin type E toxoid

Nowakowska, Maria B.; Selby, Katja; Przykopanski, Adina; Krüger, Maren; Krez, Nadja; Dorner, Brigitte G.; Dorner, Martin B.; Jin, Rongsheng; Minton, Nigel P.; Rummel, Andreas; Lindström, Miia

doi:10.1038/s41598-022-05008-1

Cited by 11 publications

(6 citation statements)

References 56 publications

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“…This has been the main barrier to elucidating a greater understanding of the pathogen. An established set of genetic tools to produce mutants in the bacteria is now available [ 321–324 ] which can allow the development of safe, non-toxigenic strains of C. botulinum, which may offer the possibility for further work outside of these select facilities in the future [ 325 ].…”

Section: Discussionmentioning

confidence: 99%

Pathogenicity and virulence of Clostridium botulinum

Rawson,

Dempster,

Humphreys

et al. 2023

Virulence

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Clostridium botulinum , a polyphyletic Gram-positive taxon of bacteria, is classified purely by their ability to produce botulinum neurotoxin (BoNT). BoNT is the primary virulence factor and the causative agent of botulism. A potentially fatal disease, botulism is classically characterized by a symmetrical descending flaccid paralysis, which is left untreated can lead to respiratory failure and death. Botulism cases are classified into three main forms dependent on the nature of intoxication; foodborne, wound and infant. The BoNT, regarded as the most potent biological substance known, is a zinc metalloprotease that specifically cleaves SNARE proteins at neuromuscular junctions, preventing exocytosis of neurotransmitters, leading to muscle paralysis. The BoNT is now used to treat numerous medical conditions caused by overactive or spastic muscles and is extensively used in the cosmetic industry due to its high specificity and the exceedingly small doses needed to exert long-lasting pharmacological effects. Additionally, the ability to form endospores is critical to the pathogenicity of the bacteria. Disease transmission is often facilitated via the metabolically dormant spores that are highly resistant to environment stresses, allowing persistence in the environment in unfavourable conditions. Infant and wound botulism infections are initiated upon germination of the spores into neurotoxin producing vegetative cells, whereas foodborne botulism is attributed to ingestion of preformed BoNT. C. botulinum is a saprophytic bacterium, thought to have evolved its potent neurotoxin to establish a source of nutrients by killing its host.

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Section: Discussionmentioning

confidence: 99%

Pathogenicity and virulence of Clostridium botulinum

Rawson,

Dempster,

Humphreys

et al. 2023

Virulence

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“…After 24 to 48 h of incubation at 30 °C or 37 °C, the cells grown in each MPN dilution series were harvested and the genomic DNA was extracted [ 50 ]. PCR detection of BoNT genes was performed as previously described using gene specific primers of bont/A [ 51 ], bont/E , bont/F [ 52 ], and bont/CD [ 53 ].…”

Section: Methodsmentioning

confidence: 99%

Specific Isolation of Clostridium botulinum Group I Cells by Phage Lysin Cell Wall Binding Domain with the Aid of S-Layer Disruption

Zhang

Douillard

Korkeala

et al. 2022

IJMS

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Clostridium botulinum is a notorious pathogen that raises health and food safety concerns by producing the potent botulinum neurotoxin and causing botulism, a potentially fatal neuroparalytic disease in humans and animals. Efficient methods for the identification and isolation of C. botulinum are warranted for laboratory diagnostics of botulism and for food safety risk assessment. The cell wall binding domains (CBD) of phage lysins are recognized by their high specificity and affinity to distinct types of bacteria, which makes them promising for the development of diagnostic tools. We previously identified CBO1751, which is the first antibotulinal phage lysin showing a lytic activity against C. botulinum Group I. In this work, we assessed the host specificity of the CBD of CBO1751 and tested its feasibility as a probe for the specific isolation of C. botulinum Group I strains. We show that the CBO1751 CBD specifically binds to C. botulinum Group I sensu lato (including C. sporogenes) strains. We also demonstrate that some C. botulinum Group I strains possess an S-layer, the disruption of which by an acid glycine treatment is required for efficient binding of the CBO1751 CBD to the cells of these strains. We further developed CBO1751 CBD-based methods using flow cytometry and magnetic separation to specifically isolate viable cells of C. botulinum Group I. These methods present potential for applications in diagnostics and risk assessment in order to control the botulism hazard.

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“…Although the vegetative cells of C. botulinum are sensitive to air, in spore form, they can retain viability for long periods. Spores of C. botulinum Group II pose the highest risk of food poisoning due to their ubiquitous presence in the environment and their ability to survive pasteurization [ 134 ], thus germinating in toxic cultures at low temperatures. Proteolytic strains can grow at temperatures below 10–12 °C (non-proteolytic—at 3–4 °C), and contaminate raw meat, fruits, vegetables, and seafood [ 135 , 136 ].…”

Section: Lab Against Bacterial Toxins and Their Producersmentioning

confidence: 99%

The Complex Role of Lactic Acid Bacteria in Food Detoxification

et al. 2022

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Toxic ingredients in food can lead to serious food-related diseases. Such compounds are bacterial toxins (Shiga-toxin, listeriolysin, Botulinum toxin), mycotoxins (aflatoxin, ochratoxin, zearalenone, fumonisin), pesticides of different classes (organochlorine, organophosphate, synthetic pyrethroids), heavy metals, and natural antinutrients such as phytates, oxalates, and cyanide-generating glycosides. The generally regarded safe (GRAS) status and long history of lactic acid bacteria (LAB) as essential ingredients of fermented foods and probiotics make them a major biological tool against a great variety of food-related toxins. This state-of-the-art review aims to summarize and discuss the data revealing the involvement of LAB in the detoxification of foods from hazardous agents of microbial and chemical nature. It is focused on the specific properties that allow LAB to counteract toxins and destroy them, as well as on the mechanisms of microbial antagonism toward toxigenic producers. Toxins of microbial origin are either adsorbed or degraded, toxic chemicals are hydrolyzed and then used as a carbon source, while heavy metals are bound and accumulated. Based on these comprehensive data, the prospects for developing new combinations of probiotic starters for food detoxification are considered.

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Construction and validation of safe Clostridium botulinum Group II surrogate strain producing inactive botulinum neurotoxin type E toxoid

Cited by 11 publications

References 56 publications

Pathogenicity and virulence of Clostridium botulinum

Pathogenicity and virulence of Clostridium botulinum

Specific Isolation of Clostridium botulinum Group I Cells by Phage Lysin Cell Wall Binding Domain with the Aid of S-Layer Disruption

The Complex Role of Lactic Acid Bacteria in Food Detoxification

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