Challenge tests reveal limited outgrowth of proteolytic Clostridium botulinum during the production of nitrate- and nitrite-free fermented sausages

Abstract

Nitrate and nitrite salts perform a versatile role in fermented meats, including the inhibition of food pathogens (in particular proteolytic group I Clostridium botulinum). Despite the increasing interest in clean-label products, little is known about the behaviour of this pathogen in response to the removal of chemical preservatives from fermented meat formulations. Therefore, challenge tests with a cocktail of nontoxigenic group I C. botulinum strains were performed to produce nitrate/nitrite-free fermented sausages under different acidification conditions and starter culture formulations, including the use of an anticlostridial Mammaliicoccus sciuri strain. Results showed limited outgrowth of C. botulinum, even in the absence of acidification. The anticlostridial starter culture did not lead to an additional inhibitory effect. The selective plating procedure adopted within this study proofed robust to follow germination and growth of C. botulinum, inhibiting common fermentative meat microbiota. The challenge tests constitute a suitable tool to assess the behaviour of this food pathogen within fermented meats upon nitrate- and nitrite omission.

Keywords: Amplicon sequencing; Challenge tests; Meat fermentation; Proteolytic Clostridium botulinum; Starter culture.