The use of commercial starter cultures in the production of Turkish semi-dry fermented sausages

 The effects of three different starter cultures (Pediococcus acidilactici, Staphylococcus xylosus plus P. pentosaceus, S. carnosus plus Lactobacillus pentosus) were evaluated during the production of Turkish semi-dry fermented sausages. Sausages were studied during the fermentation phase, after heat processing and after drying for 24 h and 72 h. Chemical and organoleptical results indicated that in the processing of these semi-dry sausages a starter culture of P. acidilactici should be used. The use of this culture significantly reduced the pH, increased the lactic acid content and percentage of total heme pigments converted to the cured pigment and improved the development of the sausages' characteristics, i.e. color, appearance, flavor and general acceptability. Received: 20 February 1998 / Revised version: 7 May 1998     

Bacterial community succession and biogenic amine changes during fermentation of fish-chili paste inoculated with different commercial starter cultures

The microbial dynamics, diversity and succession characterisation during the fermentation of fish-chili paste, with and without starter cultures by high-throughput sequencing, were investigated to identify the relationship between microbial composition and the accumulation of biogenic amines. Results showed that Firmicutes was the predominant phylum, and Lactobacillus and Weissella were the main genera during fermentation process, regardless of the inclusion of starter cultures. Compared with naturally fermented samples (NS; no starter cultures), Pediococcus and Staphylococcus flourished in samples inoculated with SBM-52 starter culture (Staphylococcus xylosus + Staphylococcus carnosus + Pediococcus pentosaceus + Pediococcus acidilactici). Staphylococcus also accounted for a larger proportion in samples inoculated with WBX-43 starter culture (Staphylococcus xylosus + Staphylococcus carnosus). Furthermore, Enterobacter was inhibited in inoculated samples. Unweighted pair-group method with arithmetic means and principal component analyses revealed that microbiota structures were different among NS, SBM and WBX samples. In case of biogenic amines, the concentration of putrescine, cadaverine and tyramine increased to higher levels in NS samples compared with inoculated samples. Our work also investigated the relationship between core bacterial communities and biogenic amines. This study provided an improved understanding for the effects of starter cultures on the microbial community and the quality of fish-chili paste.     

The application of staphylococci with flavour-generating potential is affected by acidification in fermented dry sausages

Differences in the production of bacterial metabolites with potential impact on fermented sausage flavour were found in meat simulation medium when comparing different strains of Staphylococcus xylosus and Staphylococcus carnosus as starter cultures. Overall, higher levels of 3-methyl-1-butanol and acetoin were found for S. xylosus, with some intraspecies variability. In addition, sausage fermentation parameters affected staphylococcal growth and metabolism. Strong acidification, as in Northern-European types of fermented dry sausage, inhibited S. xylosus 3PA6 but not S. carnosus 833. During a milder, Southern-European type of acidification, both strains displayed survival over time. During in situ sausage trials, variations in the degree of acidification and the choice of starter microorganisms were of importance, whereas modifications in fat and salt contents had no effects. Staphylococcus sciuri αSg2, Staphylococcus succinus 4PB1, and S. xylosus 3PA6 were unable to survive the fermentation of a Northern-European type of fermented dry sausage, characterized by low or no 3-methyl-1-butanol and acetoin production. Inoculation with S. sciuri αSg2, S. succinus 4PB1, or S. xylosus 3PA6 led to 3-methyl-1-butanol and acetoin production in Southern-European type of fermented dry sausages, which was not observed with S. carnosus 833.     

Destruction of Yersinia enterocolitica by Lactobacillus sake and Pediococcus acidilactici During Low-temperature Fermentation of Turkish Dry Sausage (sucuk)

Effects of starter cultures, Lactobacillus sake and Pediococcus acidilactici, on the survival of Yersinia enterocolitica were studied in Turkish dry sausage (sucuk). Inoculated Y. enterocolitica (approximately 5.0 log CFU/g) was eliminated completely in sausages by L. sake (about 7.0 log CFU/g) and P. acidilactici (about 7.0 log CFU/g) by the 3^rd d of fermentation but was not totally eliminated in natural fermentation without starter(s) after completion of 4 d of fermentation and 12 d drying (p < 0.05). pH decreased from 6.3 to about 4.7 in starter culture treatments and from 6.3 to 5.6 in natural fermentation after completion fermentation and drying (p < 0.05).     

Effect of processing conditions on proteolysis and taste of dry-cured sausages

 This paper deals with the influence of industrial practices such as use of starters (Lactobacillus sake and Staphylococcus carnosus), preripening (3 days at 5°C) and drying temperatures (8 and 16°C) on proteolysis, ammonia production and the taste of “salchichón”, a traditional Spanish dry-cured sausage lacking surface mould. pH dropped more sharply in the presence of a starter and at a high drying temperature (16°C); preripening did not affect the pH. The highest value for non-protein nitrogen (NPN), 16% of total nitrogen, was observed to occur in samples with the lowest pH (4.7–4.8). Total volatile basic nitrogen (TVBN) increased during drying due to the deaminase activity of internal microbial flora, which was not great (0.5–1.0% of total nitrogen) enough to affect the pH, and significantly in samples showing a larger decrease of pH. The greater proteolysis in samples with starter did not affect the assessors’ taste preference. Assessors clearly preferred those samples without starter that were dried at low temperature (8°C); however, samples with starter showed excellent commercial appearance since they dried more homogeneously. An equilibrium point must be found between acid production and taste. Received: 12 February 1996/Revised version: 24 May 1996     

Effect of processing conditions on proteolysis and taste of dry-cured sausages

 This paper deals with the influence of industrial practices such as use of starters (Lactobacillus sake and Staphylococcus carnosus), preripening (3 days at 5°C) and drying temperatures (8 and 16°C) on proteolysis, ammonia production and the taste of “salchichón”, a traditional Spanish dry-cured sausage lacking surface mould. pH dropped more sharply in the presence of a starter and at a high drying temperature (16°C); preripening did not affect the pH. The highest value for non-protein nitrogen (NPN), 16% of total nitrogen, was observed to occur in samples with the lowest pH (4.7–4.8). Total volatile basic nitrogen (TVBN) increased during drying due to the deaminase activity of internal microbial flora, which was not great (0.5–1.0% of total nitrogen) enough to affect the pH, and significantly in samples showing a larger decrease of pH. The greater proteolysis in samples with starter did not affect the assessors’ taste preference. Assessors clearly preferred those samples without starter that were dried at low temperature (8°C); however, samples with starter showed excellent commercial appearance since they dried more homogeneously. An equilibrium point must be found between acid production and taste. Received: 12 February 1996/Revised version: 24 May 1996     

The effect of different starter cultures and ripening temperatures on formation of biogenic amine in Turkish fermented sausages

Different starter culture added groups (Group I: Lactobacillus sake, Staphylococcus xylosus; Group II: Lactobacillus plantarum, Staphylococcus carnosus; Group III: Lactobacillus curvatus, Staphylococcus carnosus, and Staphylococcus xylosus) and control group sausage samples were produced experimentally at two different temperatures (22 and 26 °C). In raw materials and all groups, by the 0th, 2nd, 5th and 7th days of the ripening period, the analysis of HPLC and biogenic amine (tyramine, histamine, cadaverine, putrescine, phenylethylamine, tryptamine, spermine, and spermidine) amounts, microbiological (Lactic acid bacteria, Micrococcus/Staphylococcus, Enterobacteriaceae, Enterococcus spp., and moulds–yeasts) and chemical (pH, dry matter, and salt) have been done. During the ripening period, there is no statistical discrepancy (P > 0.05) in terms of biogenic amine amounts, microbiological, and chemical values detected from the sausage samples produced at 26 and 22 °C. However, in both levels of temperature, there is statistical discrepancy (P < 0.001) detected in terms of tyramine, putrescine values and the count of Enterococcus spp. between the starter culture added samples and control group samples. By this study, it has been stated that the ripening temperature does not make any statistical discrepancy (P > 0.05) for all values but the use of starter culture prevents the formation of biogenic amine in Turkish fermented sausages.     

Novel starter cultures to inhibit biogenic amines accumulation during fish sauce fermentation

Bacteria with amine oxidase activity have become a particular interest to reduce biogenic amines concentration in food products such as meat and fish sausages. However, little information is available regarding the application of these bacteria in fish sauce. Hence, our study was aimed to investigate the effect of such starter cultures in reducing biogenic amines accumulation during fish sauce fermentation. Staphylococcus carnosus FS19 and Bacillus amyloliquefaciens FS05 isolated from fish sauce which possess amine oxidase activity were used as starter cultures in this study. Fermentation was held for 120 days at 35 °C. The pH value increased in all samples, while salt concentration remained constant throughout fermentation. Aerobic bacteria count was significantly lower (p < 0.05) in the control than in inoculated samples as a result of starter cultures addition. However, it decreased during fermentation due to the growth inhibition by high salt concentration. Proteolytic bacterial count decreased during fermentation with no significant difference (p > 0.05) among samples. These bacteria hydrolyzed protein in anchovy to produce free amino acid precursors for amines formation by decarboxylase bacteria. The presence of biogenic amines producing bacteria in this study was considered to be indigenous from raw material or contamination during fermentation, since our cultures were negative histamine producers. Amino acid histidine, arginine, lysine and tyrosine concentration decreased at different rates during fermentation as they were converted into their respective amines. In general, biogenic amines concentration namely histamine, putrescine, cadaverine and tyramine increased throughout fermentation. However, their concentrations were markedly higher (p < 0.05) in the control (without starter cultures) as compared to the samples treated with starter cultures. Histamine concentration was reduced by 27.7% and 15.4% by Staphylococcus carnosus FS19 and Bacillus amyloliquefaciens FS05, respectively. Both cultures could also reduce other amines during fermentation. After 120 days of fermentation, the overall biogenic amines concentration was 15.9% and 12.5% less in samples inoculated with Staphylococcus carnosus FS19 and Bacillus amyloliquefaciens FS05, respectively, as compared to control samples. These findings emphasized that application of starter cultures with amines oxidase activity in fish sauce fermentation was found to be effective in reducing biogenic amines accumulation.     

The influence of environmental parameters on the catabolism of branched-chain amino acids by Staphylococcus xylosus and Staphylococcus carnosus

Degradation of the amino acids leucine, isoleucine and valine into branched flavour compounds by Staphylococcus xylosus and Staphylococcus carnosus was studied using resting cell cultures added to a defined reaction medium under different environmental conditions relevant to sausage fermentation. The environmental conditions studied were temperature (12–28°C), NaCl concentration (4.0–12.0% (w/w)) acidity (pH 4.8–5.8) and addition of manganese (0–2.5 mg Mn/kg). Flavour compounds were sampled by automatic static headspace collection and separated/quantified using gas chromatography/flame ionization detection (GC/FID). Main volatile catabolic products of leucine, isoleucine and valine were 3-methylbutanoic, 2-methylbutanoic and 2-methylpropanoic acids, respectively. The generation of branched flavour compounds was influenced significantly by most of the investigated environmental parameters, but in particularly by temperature and NaCl.Raising the temperature increased the production of branched-chain acids by both S. xylosus and S. carnosus, whereas increasing NaCl concentration had a clear negative effect, reducing the concentrations of branched-chain acids for both species but in particularly for S. carnosus. The acidity of the medium was also important for both Staphylococcus spp., but with opposite effects. Lowering pH decreased the generation of branched-chain acids by S. carnosus but increased the generation by S. xylosus. In addition, several two-factor interactions between temperature, NaCl concentration and pH were significant. Manganese addition increased the branched-chain acid concentration slightly for S. xylosus but not for S. carnosus.     

Characterization of fermented silver carp sausages inoculated with mixed starter culture

To improve the characteristics and functionality, and increase the use of fish muscle, three groups mixed starter cultures (group one: Lactobacillus plantarum-15, Staphylococcus xylosus-12 and Pediococcus pentosaceus-ATCC33316 [S-PXP]; group two: Lactobacillus planatrum-15, Staphylococcus xylosus-12 and Lactobacillus casei subs casei-1.001 [S-PXC]; and group three: Staphylococcus xylosus-12, Lactobacillus casei subsp. casei-1.001 and P. pentosaceus-ATCC33316 [S-XCP]) were inoculated in minced silver carp muscle to produce a fermented fish product. During the 48 h fermentation at 30 °C, silver carp muscle inoculated with mixed starter cultures resulted in a rapid pH decrease, suppression in the increase of thiobarturic acid (TBARS) values, total volatile base nitrogen (TVB-N), trimethylamine (TMA), and the growth of spoilage bacteria and pathogens, and had higher whiteness than the control (without any starter) (P<0.05). The changes in SDS-PAGE indicated extensive hydrolysis of muscle protein occurred during fermentation. This study showed that the mixed starter cultures could substantially improve the flavor, digestibility, and nutritional value of the silver carp muscle.     

Properties and acceptability of Som-fug, a Thai fermented fish mince, inoculated with lactic acid bacteria starters

Effect of inoculation of different lactic acid bacteria (LAB) on the fermentation and quality of Som-fug from bigeye snapper was investigated. Som-fug inoculated with Pediococcus acidilactici at 10⁴ CFU/g (PA104) had a greater acceptability than those inoculated with Lactobacillus plantarum and Pediococcus pentosaceus at either 10⁴ or 10⁶ CFU/g and the control (without inoculum). During fermentation, PA104 exhibited a higher rate of fermentation than the control as indicated by the greater rate of pH drop and lactic acid production (p<0.05). Based on pH desired (pH 4.5), the fermentation was completed within 48 and 36 h for the control and PA104, respectively. Due to higher acid production rate, the textural development of PA104 was more pronounced. Som-fug inoculated with LAB generally exhibited higher hardness and adhesiveness than the control (p<0.05), when assessed by texture profile analysis (TPA). From the result, inoculation with P. acidilactici resulted in a reduction of fermentation time and improved the quality of Som-fug. Therefore, P. acidilactici can be used as a potential starter for Som-fug fermentation.     

The effect of starter cultures on proteolytic changes and amino acid content in fermented sausages

The main objective of this work was to examine the effects of using five types of commercial starter cultures in fermented sausages. During the fermentation stage, changes in proteolytic characteristics were observed in fermented sausages. Proteolytic activity was high in Ls_b + Sc:(Lactobacillus sakei + Staphylococcus carnosus) and Pp + Sx:(Pediococcus pentosaceus + Staphylococcus xylosus) starter-inoculated sausages during processing. Moreover, a slight increase in proteolytic activity was detected during storage in both these sausages. Sarcoplasmic and myofibrillar proteins were also affected by this starter culture addition, during the fermentation, ripening and intense proteolysis were observed in both the fermented sausages. The content of free amino acids was similar at the beginning of the fermentation stage for all the studied batches. However, the high differences in the content of free amino acids at the end of the process could be attributed to the starter culture activity.     

Characterization of Micrococcaceae isolated from dry fermented sausage

One hundred Micrococcaceae strains were isolated from two types of naturally fermented dry sausages at four different stages of the ripening process in order to select the most suitable strains according to their technological characteristics including antimicrobial activity against foodborne pathogens. Identification of the isolates revealed that 91% belonged to genus Staphylococcus, whereas 5% were identified as Kocuria varians, 2% asArthrobacter agilis and 2% as Dermacoccus nishinomiyaensis. The species most often isolated was S. saprophyticus (22%), followed by S. carnosus (20%) and S. xylosus (10%). The isolated strains of S. xylosus (10 isolates), S. carnosus (20 isolates) and K. varians (five isolates) were further characterized. A large majority of the above strains was able to grow in the presence of 10% and 15% NaCl, to reduce nitrates at 18°C and to hydrolyse casein. The majority of the strains of S. xylosus produced acetoin and 30% of them hydrolysed tributyrin. All S. xylosus strains and a large majority of S. carnosus and K. varians strains exhibited an anti-listerial activity against three Listeria monocytogenes strains.The enzymatic potential of the strains was evaluated using the API ZYM system. S. xylosus strains exhibited high β -galactosidase and esterase–lipase activities, whereas S. carnosus strains indicated strong cysteine aminopeptidase and acid phosphatase activities and K. varians indicated very weak activities of the enzymes tested. A rapid characterization of the above strains according to their enzymatic pattern could be achieved in order to follow the strains selected as starter cultures during sausage fermentation. The contribution of the selected strains to a possible inhibition ofL. monocytogenes in situ on fermented meats would be of considerable interest to enhance the hygienic quality of these products.     

Effect of potassium sorbate on development of biogenic amines during sausage fermentation

Different concentrations of potassium sorbate were examined for their effect on the formation of biogenic amines during sausage fermentation using Lactobacillus plantarum and Pediococcus acidilactici as starter cultures. Microbiological examination revealed that there was a slight decrease in bacterial counts during aging period of fermented sausages. No differences in bacterial counts could be observed between treatment up to 0.06% potassium sorbate, although they were lower than that of control. Tyramine development continued, although potassium sorbate was used in concentration up to 0.06%. Histamine, tryptamine and phenylethylamine concentrations increased, then decreased in the later stage of aging to reach undetectable levels. Putrescine, cadaverine and spermidine were not detected in any sample throughout the study. Statistical analyses proved that there was a positive relation between total biogenic amine (TBA) content of fermented sausage and aging period. On the contrary, a negative relation between TBA and potassium sorbate concentration was found. The interrelationship between the concentration of TBA and the two factors (aging period and potassium sorbate concentration) with higher determination coefficient (R² = 0.94) was given.     

Survival and detection of Escherichia coli O157:H7 and Listeria monocytogenes during the manufacture of dry sausage using two different starter cultures

A sausage batter (35% pork, 35% beef, 30% fat) was inoculated with high (5·46–5·68), medium (3·78–4·54) or low (2·30–2·60 log₁₀cfug⁻¹) levels of Escherichia coli O157:H7 and with high (5·05–5·41) or medium (2·92–3·35 log₁₀cfug⁻¹) levels of Listeria monocytogenes serovar 4b and fermented using starter cultures A (Staphylococcus xylosus DD-34 with bacteriocin-producingPediococcusacidilactici PA-2 and Lactobacillus bavaricus MI-401) and B(S. carnosus MIII withLb. curvatus Lb3). Sausages were manufactured (fermented and dried) in a smoke chamber at 17–23°C for 15 days and further stored at 15–17°C for 34 days. The numbers of E. coli O157:H7 decreased more using starter B than starter A (first experiment P<0·0015, second experiment P<0·0002) but the organism was not eliminated. Small numbers of E. coli O157:H7 were more often detected after enrichment for 18–24 h than for 6 h (P=0·0044) when tested after deep freezing. By contrast, L. monocytogenes decreased more rapidly in the high-inoculum sausages produced with starter A (P<0·0001) but no significant difference was detected between the starters in the medium-inoculum sausages. L. monocytogenes was eliminated from the medium-inoculum sausages after 49 days.     

Degradation of polysaccharides and non-digestible oligosaccharides by Bacillus subtilis and Bacillus pumilus isolated from Soumbala, a fermented African locust bean (Parkia biglobosa) food Condiment

In order to select starter cultures for controlled fermentation of African locust beans, Bacillus subtilis and B. pumilus from Soumbala were investigated for their ability to degrade arabinogalactan, stachyose, raffinose and sucrose—the main polysaccharides and oligosaccharides in African locust bean. The methods used were liquefaction of gels of galactomannan for screening, pH measurement and High Performance Anion Exhange Chromatography – Pulsed Amperometric Detection (HPAEC–PAD) for detailed degradation study of the carbohydrates. For B. subtilis isolates, liquefaction of galactomannan gels was observed and HPAEC–PAD showed that arabinogalactan and stachyose were partially degraded while raffinose was fully degraded after 48 h of fermentation. Melibiose, fructose and traces of galactose were detected as residual sugars. For B. pumilus isolates much weaker degradation of the carbohydrates was observed. Two isolates of B. subtilis showed strong ability to degrade the oligosaccharides and they are seen as promising starter cultures for fermentation of African locust bean.     

Identification of Lactic Acid Bacteria and Gram‐Positive Catalase‐Positive Cocci Isolated from Naturally Fermented Sausage (Sucuk)

ABSTRACT: The aim of the study was to identify lactic acid bacteria and Gram‐positive catalase‐positive cocci isolated from Turkish dry fermented sausage (sucuk) produced by 7 different manufacturers without using starter culture. A total of 129 isolates of lactic acid bacteria were identified phenotypically. Lactobacillus plantarum was the dominant species (45.7%) followed by L. curvatus (10.9%) and L. fermentum (9.3%). Pediococcus isolates were identified as P. pentosaceus and P. acidilactici. All the isolates of gram‐positive and catalase‐positive cocci (123 isolates) were classified as Staphylococcus except for 1 isolate assigned to Kocuria rosea. The species isolated most often were S. xylosus (41.5%) and S. saprophyticus (28.5%). Four isolates were identified as S. equorum (3.3%), 1 isolate was assigned to S. carnosus (0.8%).     

Starter Culture and Time/Temperature of Storage Influences on Quality of Fermented Mutton Sausage

Rate of acid production and ability to produce antimicrobial activity were tested on P. acidilactici H and L. plantarum 27 during growth in a sausage formula mixture. Lactacel 75 was used as control. Starter culture did not affect water activity, pH, ash or protein content, fatty acid composition (except for nonadecanoate), cholesterol content or retention of moisture, protein, fat, ash or fatty acids. Sausages made with P. acidilactici H had the lowest (P<0.05) retention (89.6%) of cholesterol during processing and storage. Sensory evaluation of fermented sausage and bologna prepared from the two starter cultures or from Lactacel 75 indicated both products were acceptable after 50–60 days storage at 4°C.     

Effect of starter culture on proteolytic changes during processing of fermented beef sausages

Fermented beef sausages inoculated with four different starter cultures (Pediococcus acidilactici,Lactobacillus curvatus, Lactobacillus sake, or Streptomyces griseus) were evaluated for proteolysis during process stages (prefermentation, fermentation, drying and heating). Increases (p ? 0.05) in the nonprotein nitrogen (NPN) fraction were found at sequential stages of processing, while starter cultures had no major effects on NPN content. Concentrations of most free amino acids increased (p ? 0.05) during fermentation and drying, and culture effects were found for differences among concentrations of some individual free amino acids. From SDS–PAGE analysis of sarcoplasmic and myofibrillar protein fractions after fermentation and drying, myosin heavy and light chains, actin and troponin were degraded during processing. However, starter culture effects were absent from SDS–PAGE protein patterns.     

Fermentation of bighead carp (Aristichthys nobilis) surimi and the characteristics of fermented bighead carp surimi products

BACKGROUND: To improve the properties and functionality of bighead carp (Aristichthys nobilis) muscle, bighead carp surimi was inoculated with combinations of Saccharomyces cerevisiae, Lactobacillus and Monascus and fermented, and then bighead carp sausages were prepared. The characteristics of fermented bighead carp surimi and sausages were investigated. RESULTS: During the 24 h fermentation at 30 °C, bighead carp surimi inoculated with mixed starter cultures resulted in a rapid decrease of pH, and suppression in the growth of Enterobacteria. The bighead carp sausages exhibited better colour, taste, flavour and appearance than the control (P < 0.05). The changes in non‐protein nitrogen (NPN), free amino acid in the fermented bighead carp surimi and SDS–PAGE patterns indicated that severe hydrolysis of muscle protein had occurred during fermentation. The sausages inoculated with the mixed starter cultures gained higher scores for flavour and overall acceptability than the control. CONCLUSIONS: The characteristic of fermented bighead carp surimi and sausages can be significantly improved by fermentation with the combinations of yeast, Lactobacillus and Monascus. Copyright © 2008 Society of Chemical Industry