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Browsing by Author "Laakso, Kristina"

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  • Laakso, Kristina (2019)
    Clostridium botulinum is a Gram-positive, anaerobic, spore-forming bacterium that is found widely in nature. C. botulinum produces highly potent neurotoxin which causes paralysis. Yet, it is not known why the toxin is produced. C. botulinum poses a risk for the food industry, when spores germinate in food and start producing toxin. The aim of this study was to report how different carbohydrates and metabolites affect growth, toxin production and sporulation of C. botulinum. The hypothesis was that different substrates have different influence on the metabolism of C. botulinum. Earlier studies show which carbohydrates are utilized by C. botulinum but only few relate nutrient availability to toxin production. Glucose is the far most studied carbohydrate and it is known to support growth and toxin production. Growth in defined medium with added substrates was measured with Bioscreen. The method is based on measuring optical density of the cultures; optical density increases when the bacteria divide. Toxin levels were measured from 1 d and 5 d samples with a commercial ELISA (ELISA, enzyme-linked-immunosorbent-assay). The number of spores produced was measured after five days of growth. Vegetative cells were destroyed by heating (10 min, 80 °C) and MPN-method was conducted (MPN, most-probable number). With added glucose and glucose derivates (trehalose, maltotriose), growth and toxin production were induced most compared to control medium. High spore numbers were also measured. Higher concentrations of these substrates supported growth more than the lower concentrations. With higher concentrations of glucose and maltotriose the toxin levels were lower compared to low concentrations. With trehalose toxin levels were similar at high and low concentration. It can be concluded that the maximal growth density alone does not determine the toxin levels. N-acetylglucosamine (GlcNAc), which is found in bacterial cell walls, supported growth similar to glucose. GlcNAc supported also high sporulation and toxin production. Even though GlcNAc is a major component of chitin (e.g. the insect exoskeleton), chitin did not support toxin production or sporulation as efficiently as GlcNAc. The results indicate that C. botulinum might favor environments with fungi, dead bacteria or degraded chitin. By utilizing compounds from dead bacteria or insects, C. botulinum might have a role in circulating nutrients in anaerobic environment.