Prevalence of Listeria monocytogenes in fresh and fermented Italian sausages and ribotyping of contaminating strains

Listeria monocytogenes has been detected in fresh as well as dry and semidry fermented sausages, rendering preparation and consumption of these products as a potential risk to human health. The aims of this study were (1) to evaluate the L. monocytogenes prevalence in 288 fresh and 237 fermented sausages produced in northern Italy; (2) to quantify the average pathogen Most Probable Number (MPN) per g of sausage; (3) to evaluate the sausage strain genetic diversity by automated PvuII ribotyping; and (4) to predict the pathogenicity lineage of these isolates determining their DuPont Identification Library Codes (DUP-IDs) by EcoRI ribotyping. The overall prevalence of L. monocytogenes in the sampled sausages was 28.2%. The percentage of L. monocytogenes positive fresh sausages was significantly higher than that of fermented sausages (i.e. 38.9 vs 15.2%), which had a pathogen load always lower than 10 MPN/g. In contrast, 16.1% of fresh sausages were contaminated by 10 to 100 MPN/g and 20.5% had more than 100 MPN/g. PvuII successfully discriminated sausage isolates with a Simpson's numerical index of discrimination of 0.637. A total of 12 and 9 different PvuII ribogroups were identified among 47 fresh and 24 fermented randomly selected sausage strains, respectively. Six of those ribogroups were shared between strains contaminating both kinds of sausages. According to the evaluation of the strain DUP-IDs, the majority of the isolates investigated in this study were part of the type II L. monocytogenes pathogenicity lineage, but type I lineage strains were identified among fermented sausage isolates. In conclusion, L. monocytogenes prevalence in Italian sausages was estimated to be around 28.2%. However, 84.2% of the samples were contaminated by less than 100 MPN of L. monocytogenes per g and the majority of L. monocytogenes contaminating strains would be classified in the type II pathogenicity lineage, including serotypes 1/2a, 1/2c and 3a.     

Characterisation of naturally fermented sausages produced in the North East of Italy

In the Friuli Venezia Giulia region, in the North East of Italy, a traditional fermented sausage is produced without the use of microbial starters. It is characterized at the end of the ripening period by accentuated acidity, slight sourness and elastic, semi-hard consistency. In this study, three fermentations, carried out in different seasons (winter, spring and summer) were followed analyzing the microbiological, physicochemical and sensory aspects of this product. The sausages were characterized by an important microbial activity of lactic acid bacteria and micro/staphylococci that resulted in a product with a final pH of about 5.6-5.7. An interesting aspect was the high number of fecal enterococci that can play an important role in the definition of the organoleptic profile of the final product. No Listeria monocytogenes, Salmonella spp. and Staphylococcus aureus were ever isolated from the raw materials or the fermented sausages during the maturation, underlining the safety of this product. The final water activity of the product was 0.91-0.92. One hundred and fifty lactic acid bacteria were isolated and identified by molecular methods to understand which species were more predominant in the product. Lactobacillus curvatus and Lactobacillus sakei were the most numerous (54 and 64 strains isolated, respectively) and they were the only species common to all three fermentations. A cluster analysis of the profiles obtained from these strains after RAPD-PCR highlighted a population distribution that was fermentation-specific.     

Characterization of lactic acid bacteria isolated from a Greek dry-fermented sausage in respect of their technological and probiotic properties

A total of 147 lactic acid bacteria was isolated from two types of naturally fermented dry sausages at four different stages of the ripening process studied in order to select the most suitable strains according to their technological characteristics including probiotic properties and antimicrobial activity against food-borne pathogens. Identification of the isolates revealed that 90% were lactobacilli, 4% enterococci, 3% Pediococcus sp. and sporadic isolates of Weissella viridescens, Leuconostoc pseudomesenteroides, and Leuconostoc sp. The isolated strains of Lactobacillus sakei (49 isolates), Lactobacillus curvatus (24 isolates) and Lactobacillus plantarum (7 isolates) were further characterized. All strains could grow at 15?°C, whereas the majority of the strains was able to grow in the presence of 6.5% NaCl and on acetate agar. The enzymatic potential of the strains was evaluated using the API ZYM system. During in vitro investigations all strains exhibited high leucine and valine aminopeptidase activities and moderate acid phosphatase and phosphohydrolase activities. Some strains showed very weak lipolytic activity. The enzyme profiling is an important factor for selection of strains as starter cultures. A large majority of the strains tolerated 0.1% bile salts whereas 58% of Lactobacillus curvatus strains and all Lactobacillus plantarum strains were resistant to 0.3% bile salts. All Lactobacillus sakei strains and the majority of Lactobacillus curvatus and Lactobacillus plantarum strains exhibited an anti-listerial activity against three Listeria monocytogenes strains. A percentage of 75, 50 and 29% of Lactobacillus sakei, L. curvatus and L. plantarum strains, respectively, could inhibit two Staphylococcus aureus strains. The contribution of the selected strains to a possible inhibition of Listeria monocytogenes and S. aureus in situ on fermented meats would be of considerable interest to enhance the hygienic quality of these products.     

Phenotypic and technological diversity of lactic acid bacteria and staphylococci isolated from traditionally fermented sausages in southern Greece

The physicochemical and microbiological characteristics of spontaneously fermented sausages made by two medium-sized enterprises (MSE) located in southern Greece have been studied. A total of 300 lactic acid bacteria and 300 staphylococcal strains have been isolated and identified by their physiological characteristics. Lactobacillus plantarum strains were found to dominate the lactic acid bacteria microbiota in most of the cases with L. sakei strains prevailing in some of them and L. rhamnosus strains occasionally accompanying the dominant lactic acid bacteria microbiota. On the other hand, S. saprophyticus strains were found to dominate the staphylococcal microbiota in all spontaneously fermented sausages with of S. simulans, S. xylosus, S. gallinarum and S. cohnii cohnii strains being sporadically present. Following the identification, an evaluation of their technological properties, namely proteolytic and lipolytic capacities as well as production of biogenic amines and antimicrobial compounds, took place. None of the lactic acid bacteria and staphylococci was found to possess lipolytic activity whereas a total of 6 lactic acid bacteria and 51 staphylococci strains were found to be able to hydrolyse either the sarcoplasic, myofibrillar or both protein fractions. Furthermore, only one L. sakei strain and 185 staphylococci strains were found to possess decarboxylase activity against lysine, tyrosine, ornithine or histidine. Finally none of the staphylococcal microbiota and 3 lactic acid bacteria strains were found to be able to produce antimicrobial compounds of proteinaceous nature against Listeria monocytogenes.     

Use of bacterial extracts to enhance amino acid breakdown in dry fermented sausages

The effect of the intracellular cell-free extracts (ICFEs) of two bacterial strains (Lactobacillus sakei GO and Bacillus pumilus) on the amino acid catabolism and the sensory properties of dry fermented sausages, was investigated. Extracts were added to sausages alone or in combination with a protease, papain. Amino acid breakdown was monitored by the changes in free amino acids, ammonia and amine content during the ripening process. A 15% decrease in the content of free amino acids was observed in sausages added with the ICFE from L. sakei GO. Furthermore, the extract of L. sakei GO significantly reduced (54-68%) the content of the amino acids considered as precursors of the typical ripened flavour, i.e., valine, leucine and isoleucine. Chemical changes were not reflected in a significant improvement of the sensory quality of sausages added with the ICFEs. The potential use of the bacterial ICFEs studied in the present work for the manufacture of dry fermented sausages, and its comparison with the use of fungal extracts, are discussed.     

Starter cultures and high-pressure processing to improve the hygiene and safety of slightly fermented sausages

The effectiveness of selected starter cultures and high hydrostatic pressure after ripening was evaluated to improve the safety and quality of slightly fermented sausages. Inhibition of common foodborne pathogens, spoilage bacteria, and biogenic amine content was studied. Random amplification of polymorphic DNA and plasmid profiles were used to monitor the competitiveness of the starter cultures during fermentation and ripening. Lactobacillus sakei CTC6626 and Staphylococcus xylosus CTC6013 dominated L. sakei CTC6469 and S. xylosus CTC6169 independently of the product assayed. Starter cultures were able to control the growth of Listeria monocytogenes, Enterobacteriaceae, Enterococcus, and the biogenic amine content. A pH decrease below 5.3 at the seventh day of fermentation was crucial. Salmonella spp. counts decreased significantly during ripening independently of the use of starter culture and product. High hydrostatic pressure treatment was necessary to ensure absence of Salmonella spp. in final products.     

Safety improvement and preservation of typical sensory qualities of traditional dry fermented sausages using autochthonous starter cultures

Traditional dry fermented sausages are manufactured without addition of starter cultures in small-scale processing units, their fermentation relying on indigenous microflora. Characterisation and control of these specific bacteria are essential for the sensory quality and the safety of the sausages. The aim of this study was to develop an autochthonous starter culture that improves safety while preserving the typical sensory characteristics of traditional sausages. An autochthonous starter composed of Lactobacillus sakei, Staphylococcus equorum and Staphylococcus succinus isolated from a traditional fermented sausage was developed. These strains were tested for their susceptibility to antibiotics and their production of biogenic amines. This starter was evaluated in situ at the French traditional processing unit where the strains had been isolated. Effects of the autochthonous starter were assessed by analysing the microbial, physico-chemical, biochemical and sensory characteristics of the sausages. Inoculation with the chosen species was confirmed using known species-specific PCR assays for L. sakei and S. equorum and a species-specific PCR assay developed in this study for S. succinus. Strains were monitored by pulse-field gel electrophoresis typing. Addition of autochthonous microbial starter cultures improved safety compared with the traditional natural fermentation of sausages, by inhibiting the pathogen Listeria monocytogenes, decreasing the level of biogenic amines and by limiting fatty acid and cholesterol oxidation. Moreover, autochthonous starter did not affect the typical sensory quality of the traditional sausages. This is the first time to our knowledge that selection, development and validation in situ of autochthonous starter cultures have been carried out, and also the first time that S. equorum together with S. succinus have been used as starter cultures for meat fermentation. Use of autochthonous starter cultures is an effective tool for limiting the formation of unsafe compounds in traditional sausage while preserving their original and specific sensory quality.     

Fate of Listeria monocytogenes in experimentally contaminated French sausages

Listeria monocytogenes has been recognized as one of the most important foodborne pathogens dealt with by the food. The bacterium has been found in every part along the pork processing industry from the slaughterhouse to the cutting room and the delicatessen factories. During the fermentation and drying of sausages, L. monocytogenes tends to decrease substantially. However, despite the various hurdles in the dry sausage manufacturing process, L. monocytogenes is able to survive and is detected in the final products. The present study has evaluated growth and survival of eight different L. monocytogenes strains (originating from sausage, sausage industry environment and from clinical cases of listeriosis) in experimentally inoculated French sausages with 10(4) cfu g(-1). This study points out the fact that the decrease of L. monocytogenes contamination rate during the manufacturing process of sausages is strain dependent (p < 0.001) and mainly due to the drying and maturation step than to the fermentation itself. Whatever the strains studied, almost no decrease of the contamination rate was noted during the fermentation step. However hurdle-adapted strains (those isolated from sausages or sausage industry environment) were more difficult to cure from sausages (decrease by 1.5 log10) than non-adapted strains (decrease by 3 log10) at the end of the drying period (day 35), when sausages were ready for consumption. These sausages became safe only at the best before date. As a consequence, L. monocytogenes and more particularly those "adapted" strains might represent a very important issue for hygienists since these strains originating from sausages or production environment themselves are likely to contaminate sausages during manufacturing and remain in the final products. However, the high inoculum levels used in the study (10(4) cfu g(-1)) are not representative of the natural contamination of L. monocytogenes commonly encountered in the raw material for sausages. If such contamination happened to be inferior to 100 cfu g(-1), then the manufacturing process used in this study would be able to produce "safe" sausages according to the European regulation requiring the absence of L. monocytogenes in 25 g of food with a tolerance of below 100 cfu g(-1) at the best before date.     

Survival of Listeria monocytogenes strains in a dry sausage model

The survival of five inoculated Listeria monocytogenes strains (DCS 31, DCS 184, AT3E, HT4E, and HR5E) was studied in dry fermented sausages prepared using two different starter cultures (starter A and B) with or without a protective Lactobacillus plantarum DDEN 2205 strain. L. monocytogenes was detected throughout ripening in every sausage sample in which the L. plantarum DDEN 2205 strain had not been used. The use of either starter A, with a high concentration of protective culture, or starter B, with a low concentration of protective culture, resulted in L. monocytogenes-negative sausages after 17 days of ripening. Differential survival was noted among the L. monocytogenes strains during fermentation. Strains AT3E and DCS 31 survived in sausages with protective cultures more often than did the other strains, whereas HT4E and HR5E were inhibited during ripening by all starter and protective cultures used. Protective cultures such as L. plantarum may be used as part of a hurdle strategy in dry sausage processing, but variations in susceptibility of different L. monocytogenes strains can create problems if other hurdles are not included.     

Mixed starter cultures to control biogenic amine production in dry fermented sausages

Several combinations of an amine-negative Lactobacillus sakei strain, along with proteolytic Staphylococcus carnosus or Staphylococcus xylosus strains, were used to study the influence of mixed starter cultures on biogenic amine production during the manufacture of dry fermented sausages. Changes in pH, water content, proteolysis, microbial counts, and biogenic amine contents were simultaneously examined in a spontaneously fermented batch and in three mixed starter-mediated batches. A double-controlled microbial charge initially inoculated as mixed starter culture of L. sakei and Staphylococcus spp. (all amine-negative strains) drastically reduced tyramine, cadaverine, and putrescine accumulation. No production of other aromatic amines such as histamine, phenylethylamine, or tryptamine was observed in any batch. The polyamines, spermine and spermidine, were found in raw materials and their levels decreased slightly in the spontaneously fermented batch. No correlation between proteolysis and biogenic amine production was observed. The use of proper technological conditions favoring starter development and the use of the raw materials with good hygienic quality make it possible to produce fermented sausages nearly free of biogenic amines.     

Prevalence of Salmonella spp. and Listeria monocytogenes at small-scale spanish factories producing traditional fermented sausages

The manufacturing of fermented sausages is subject to natural contamination processes that can potentially carry foodborne pathogens along the process chain and result in contamination of the final product. The aim of this study was to evaluate the occurrence of Salmonella spp. and Listeria monocytogenes at different sampling points during the manufacturing process of fuet, a type of traditional fermented sausage, at 10 small-scale Spanish factories. The presence of both pathogens was studied in the raw materials (19 casings and 19 meat batters), the final products (19 fermented sausages), and the factory equipment (12 mincing, 12 mixing, and 19 stuffing machines, 19 cutting tables, 11 knives, and 12 cold rooms) by using classical microbiological techniques and real-time PCR. Salmonella was not detected in the equipment analyzed or in the final products, but it was detected in the raw materials (23.7% of samples). L. monocytogenes showed higher incidence than Salmonella and was detected in the equipment (11.8% of samples), the raw materials (28.9%), and the final products (15.8%), confirming its ubiquity throughout the manufacturing process of fermented sausages. Five factories were further investigated to study the changes in the distribution of pathogens in the fuet production process over a period of either 2 or 3 years. There was considerable variation in the incidence of both pathogens at different sampling periods, and there was no relation between seasonal variations or geographic location of the factories.     

Diversity of microorganisms in the environment and dry fermented sausages of small traditional French processing units

Naturally fermented sausages produced in nine traditional French processing units and their environmental surfaces were characterised by microbial and physico-chemical analyses. Salmonella and Staphylococcus aureus were not detected in the environment whereas Listeria monocytogenes was detected in four samples. Staphylococcus/Kocuria, lactic acid bacteria (LAB), Enterobacteriaceae, Pseudomonas, yeasts/moulds and enterococci contaminated the surfaces of two processing units, indicating insufficient cleaning and disinfection procedures. The final sausages did not present any health risk in seven of the processing units. In two of the processing units, the final sausages were contaminated with S. aureus and L. monocytogenes, respectively, at levels exceeding the maximum tolerable limit. Staphylococcus/Kocuria and LAB grew well in the products. Biogenic amines were found in the majority of the final products. Their occurrence was associated with high numbers of lactic acid bacteria and enterococci. The study outlined the processing and microbial diversities of French naturally fermented sausages.     

Proteolytic and lipolytic starter cultures and their effect on traditional fermented sausages ripening and sensory traits

In this study, three starter formulations including Lactobacillus curvatus and Staphylococcus xylosus strains selected in vitro on the basis of their lipolytic and proteolytic activities were employed for the manufacture of traditional fermented sausages of southern Italy. Microbial population, proteolysis, lipolysis, changes in free amino acids (FAA) and free fatty acids (FFA) and development of characteristic taste and flavor of the final product were investigated. Proteolysis and lipolysis were observed in sausages inoculated with proteolytic and lipolytic S. xylosus coupled with L. curvatus, while the sausage started with only S. xylosus without lactobacilli was identical to the non-inoculated control, indicating that the proteolysis could be due to both microbial activity and endogenous proteases activated by the decrease in pH. The statistical analysis applied to the instrumental and sensory data showed that there was an effect of the starter used on the characteristics of the sausage obtained. In particular, the control samples showed very close features different from the sausages obtained by adding starter cultures. Finally, analyzing the sensory parameters the sausages ripened without starter addition and those started without the L. curvatus AVL3 showed similar features indicating an influence of the presence of the lactobacilli on the final organoleptic quality of the sausages. An appropriate choice of a combination of strains in a starter formulation is fundamental to obtain products of the expected quality.     

Biodiversity and dynamics of meat fermentations: the contribution of molecular methods for a better comprehension of a complex ecosystem

The ecology of fermented sausages is complex and includes different species and strains of bacteria, yeasts and molds. The developments in the field of molecular biology, allowed for new methods to become available, which could be applied to better understand dynamics and diversity of the microorganisms involved in the production of sausages. Methods, such as denaturing gradient gel electrophoresis (DGGE), employed as a culture-independent approach, allow to define the microbial dynamics during the fermentation and ripening. Such approach has highlighted that two main species of lactic acid bacteria, namely Lactobacillus sakei and Lb. curvatus, are involved in the transformation process and that they are accompanied by Staphylococcus xylosus, as representative of the coagulase-negative cocci. These findings were repeatedly confirmed in different regions of the world, mainly in the Mediterranean countries where dry fermented sausages have a long tradition and history. The application of molecular methods for the identification and characterization of isolated strains from fermentations highlighted a high degree of diversity within the species mentioned above, underlining the need to better follow strain dynamics during the transformation process. While there is an important number of papers dealing with bacterial ecology by using molecular methods, studies on mycobiota of fermented sausages are just a few. This review reports on how the application of molecular methods made possible a better comprehension of the sausage fermentations, opening up new fields of research that in the near future will allow to unravel the connection between sensory properties and co-presence of multiple strains of the same species.     

Sensory improvement of dry-fermented sausages by the addition of cell-free extracts from Debaryomyces hansenii and Lactobacillus sakei

The effects of the addition of a combined cell-free extract from Lactobacillus sakei and Debaryomyces hansenii (D+L) or just a D. hansenii cell-free extract (D) to the initial formulation of a dry-fermented sausage were evaluated. The differences found among batches in the main microbial populations, pH, moisture content and global proteolytic and lipolytic indexes (total free amino acids, non protein nitrogen, acidity and tiobarbituric acid index) were not significant. Only, the acidity value of batch D was significantly higher (p<0.05) than that of batch D+L. Thus, cell-free extract from D. hansenii accelerated the lipolysis. Moreover, there were some significant differences (p<0.05) in the amino acid profile and, especially, in the aroma profile. The combination D+L and D promoted the generation of volatile compounds derived from lipid oxidation and carbohydrate fermentation. In batch D, the production of volatile compounds derived from amino acid catabolism and microbial fermentation was also enhanced. The overall quality was improved by both treatments (D+L, D) and also the aroma by addition of the combination of extracts (D+L). It is concluded that the addition of cell-free extracts from D. hansenii and, particularly, D. hansenii plus L. sakei could be useful to improve the final quality of fermented sausages.     

Effect of packaging and storage temperature on the survival of Listeria monocytogenes inoculated postprocessing on sliced salami

The survival of postprocess Listeria monocytogenes contamination on sliced salami, stored under the temperatures associated with retail and domestic storage, was investigated. Sliced salami was inoculated with low and high concentrations of L. monocytogenes before being packaged under vacuum or air. Survival of L. monocytogenes was determined after storage of sausages for 45 or 90 days for low or high sample inocula, respectively, at 5, 15, and 25 degrees C. All survival curves of L. monocytogenes were characterized by an initial rapid inactivation within the first days of storage, followed by a second, slower inactivation phase or "tailing." Greater reduction of L. monocytogenes was observed at the high storage temperature (25 degrees C), followed by ambient (15 degrees C) and chill (5 degrees C) storage conditions. Moreover, vacuum packaging resulted in a slower destruction of L. monocytogenes than air packaging, and this effect increased as storage temperature decreased. Although L. monocytogenes numbers decreased to undetectable levels by the end of the storage period, the time (in days) needed for this reduction and for the total elimination of the pathogen decreased with high temperature, aerobic storage, and high inoculum. Results of this study clearly indicated that the kinetics of L. monocytogenes were highly dependent on the interaction of factors such as storage temperature, packaging conditions, and initial level of contamination (inoculum). These results may contribute to the exposure assessment of quantitative microbial risk assessment and to the establishment of storage-packaging recommendations of fermented sausages.     

Competitiveness and antibacterial potential of bacteriocin-producing starter cultures in different types of fermented sausages

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.     

EVALUATION OF FUNCTIONAL ASPECTS IN LACTOBACILLUS STRAINS ISOLATED FROM DRY FERMENTED SAUSAGES

ABSTRACT

The aim of this study was to characterize lactobacilli isolated from a traditional Italian dry‐fermented sausages inoculated with Lactobacillus casei/paracasei for use as probiotics. Strains isolated were L. casei (38%), L. paracasei paracasei (32%), Lactobacillus rhamnosus (21%) and Lactobacillus sakei sakei (9%). L. paracasei paracasei and L. rhamnosus showed no significant decrease in viability compared with L. casei and L. sakei sakei in artificial gastric juice; L. casei and L. sakei sakei were not affected by artificial intestinal fluid. In human embryonic intestine cell line (INT407) in vitro model, L. rhamnosus and L. sakei sakei were more adhesive than the other strains. The production of acetic acid (40.92 ± 1.94 mmol/L) and propionic acid (0.14 ± 0.07 mmol/L) was similar in all strains tested. Considerably higher (100–250 mmol/L) levels of lactic acid were produced by L. casei, L. paracasei paracasei and L. rhamnosus, and were varied depending on microaerobic or anaerobic conditions. L. casei and L. rhamnosus had the best antibacterial activity against Escherichia coli and Salmonella enterica ssp. enterica (serovar Typhimurium). These findings suggest that L. casei and L. rhamnosus merit further investigation as novel starter cultures in meat products.

PRACTICAL APPLICATIONS

Probiotics are well established in dairy products but they are not widely used in meat products and it is expected their use in this area will expand considerably in the near future. Starter cultures of active or resting forms of microorganisms are widely employed in fermented meat products to improve sensory qualities. More recently they are being used in fermented meat products to improve the nutritional value of these products as functional foods. Probiotics in meat may exert their benefits by several methods, and more research is required to identify the strains and combinations of strains that produce the most health benefits. Our study demonstrates that the in vitro characterization of Lactobacillus strains directly isolated from dry fermented sausages can be useful to select novel starter cultures with potential probiotic characteristics among lactic acid bacteria used as commercial meat starter culture.

     

Effectiveness of a Lactobacillus sakei starter culture in the reduction of biogenic amine accumulation as a function of the raw material quality

The effectiveness of an amine-negative starter culture (Lactobacillus sakei CTC494) in the reduction of biogenic amine production during the ripening of fermented sausages was examined. Four batches were manufactured in parallel: spontaneously fermented and starter-mediated sausages were manufactured from two lots of raw materials of different hygienic quality. Besides the biogenic amine contents, changes in the microbial counts, nitrogenous fractions, pH, and water content were measured at several sampling points during the ripening process. In sausages manufactured from good quality meat, the starter strain of L. sakei reduced and even inhibited biogenic amine accumulation during sausage fermentation, the end products showing extremely low biogenic amine contents (tyramine levels less than 15 mg/kg of dry matter and putrescine and cadaverine levels less than 5 mg/kg of dry matter). Nevertheless, starter-mediated sausages made from poorer-quality raw materials showed much higher amine contents (308, 223, and 36 mg/kg of dry matter of cadaverine, tyramine, and putrescine, respectively), which were only slightly lower than those of the spontaneously fermented sausages made from the same raw materials. The relatively high bacterial numbers of raw materials of poorer-hygienic quality diminished the beneficial effect of the starter strain. Therefore, the effectiveness of the starter was strongly dependent on the hygienic quality of the raw materials used.     

Industrial application of an antilisterial strain of Lactobacillus sakei as a protective culture and its effect on the sensory acceptability of cooked, sliced, vacuum-packaged meats.

The application of a protective lactic acid bacterium (LAB) during the commercial production of cooked meat products is described. The LAB, a strain of Lactobacillus sakei, was previously isolated from cooked ham and inhibited growth of Listeria monocytogenes and Escherichia coli O157:H7 in this product. L. sakei was applied to the cooked products at a concentration of 10(5)-10(6) cfu/g immediately before slicing and vacuum-packaging using a hand-operated spraying bottle. The LAB strain inhibited growth of 10(3) cfu/g of a cocktail of three rifampicin resistant mutant L. monocytogenes strains both at 8 degrees C and 4 degrees C. Consumer acceptance tests of cooked ham and of servelat sausage, a Norwegian non-fermented cooked meat sausage, showed that control and inoculated products were equally acceptable. The products were still acceptable after storage for 28 days at 4 degrees C and, after opening the packages, for a further 5 days at 4 degrees C. The findings presented here confirm that the L. sakei strain is suitable for use as a protective culture and may technically easily be implemented in the commercial production of cooked meat products.