Application of bacteriocin-producing starter cultures of lactic acid bacteria in fermented sausage production contributes to food safety. This is sometimes hampered by limited efficacy in situ and by uncertainty about strain dependency and universal applicability for different sausage types. In the present study, a promising antilisterial-bacteriocin producer, Lactobacillus sakei CTC 494, was applied as a coculture in addition to commercial fermentative starters in different types of dry-fermented sausages. The strain was successful in both Belgian-type sausage and Italian salami that were artificially contaminated with about 3.5 log CFU g(-1) of Listeria monocytogenes. After completion of the production process, this led to listerial reductions of up to 1.4 and 0.6 log CFU g(-1), respectively. In a control sausage, containing only the commercial fermentative starter, the reduction was limited to 0.8 log CFU g(-1) for the Belgian-type recipe, where pH decreased from 5.9 to 4.9, whereas an increase of 0.2 log CFU g(-1) was observed for Italian salami, in which the pH rose from 5.7 to 5.9 after an initial decrease to pH 5.3. In a Cacciatore recipe inoculated with 5.5 log CFU g(-1) of L. monocytogenes and in the presence of L. sakei CTC 494, there was a listerial reduction of 1.8 log CFU g(-1) at the end of the production process. This was superior to the effect obtained with the control sausage (0.8 log CFU g(-1)). Two commercial antilisterial cultures yielded reductions of 1.2 and 1.5 log CFU g(-1). Moreover, repetitive DNA sequence-based PCR fingerprinting demonstrated the competitive superiority of L. sakei CTC 494.
The objective of this study was to investigate Clostridium botulinum growth and toxin production in the industrially manufactured Italian Parma ham. The study focuses on the Parma ham production phase identified as maximum risk to C. botulinum proliferation, i.e. the transition from cold phase (salting and resting) to a phase carried out at temperature between 15 and 23°C (drying). A preliminary in vitro test was carried out in order to verify the capability of 6 C. botulinum strains (1 type A, 4 type B, and 1 type E strains) to grow in conditions of temperature, pH and NaCl concentration comparable to those of the beginning stage of ham drying. Five C. botulinum strains grew at 20°C and pH 6, four strains produced toxin when inoculated at a concentration equal to 103 cfu/mL at NaCl concentration of 4%, while when the inoculum concentration was 10 cfu/mL, NaCl concentration of 3% resulted the toxin-genesis limiting factor. An experimental contamination with a mixture of the 5 C. botulinum strains selected by the preliminary in vitro test was performed on 9 thighs inoculated at the end of the resting phase. The study was designed to evaluate the potential growth and toxin production in extremely favourable conditions for the bacterium. Type B proteolytic C. botulinum toxin was produced after 14 days of incubation at 20°C in 2 thighs characterised by high weight, low number of days of resting and anomalous physiochemical characteristics [one for very low NaCl concentration (1.59%), the other for elevated pH (6.27) and both for high water activity values (>0.970)]. The results of this research confirm that the cold resting step is a critical phase in the production process of Parma ham for the investigated hazard. Based on the present study, the long resting phase adopted in the manufacturing of Parma ham is proven effective to prevent the growth of C. botulinum, an event which could not otherwise be excluded if the hams were processed under less stringent technological conditions.
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