Use of autochthonous Pediococcus acidilactici and Staphylococcus vitulus starter cultures in the production of "chorizo" in 2 different traditional industries

The present study determined how the different ripening conditions affected the growth and development of 3 autochthonous starter cultures, and the physico-chemical and sensory characteristics of chorizo. Each of 3 strains of Pediococcus acidilactici (MC184, MS198, and MS200) and one of Staphylococcus vitulus (RS34) were associated to prepare the starter cultures, P184S34, P198S34, and P200S34. Then, chorizo was prepared following 2 manufacturing procedures. The autochthonous starter cultures were able to compete and colonize the sausages in both ripening procedures. The use of the starter cultures showed evident differences by the texture analysis, with the control batches being generally tougher than the starter culture batches. Also, the highest biogenic amine (BA) levels were found in control batches and the lowest in P200S34 batches. While the use of these starter cultures does not change the sensory characteristics of these traditional fermented sausages, it improves their homogeneity and safety, except for P184S34 batch in which more BAs are detected in industry 2. PRACTICAL APPLICATION: The 3 autochthonous starter cultures selected could be used in traditional industries because they are able to compete well and colonize the dry fermented sausages "chorizo." The use of these starter cultures improves the texture and homogeneity of traditional fermented sausages. Biogenic amines decreased in the starter cultures batches improving the safety.     

Influence of starter cultures on the generation of antioxidant nitrogen compounds in Iberian dry‐fermented sausages

The purpose of this work was to investigate the influence of a starter culture on the generation of nitrogen compounds with antioxidant activity during the ripening of Iberian dry‐fermented sausages. Starters P200S34 (P. acidilactici M200 and S. vitulus RS34) and P198S34 (P. acidilactici MS198 and S. vitulus RS34) were used to make the Iberian dry‐fermented sausages ‘salchichón’ and chorizo; then, the physicochemical and microbial properties were determined during the ripening process. The oxygen radical absorbance capacity (ORAC) assay was employed to evaluate the antioxidant potential of the nitrogen extracts obtained during ripening of the sausages. This activity was correlated with the most relevant compounds detected by HPLC‐ESI‐MS in the final ripened extracts. Although a relevant part of the antioxidant activity was attributed to the predominant natural nitrogen fraction, the microbial population found in fermented sausages and the fermentation conditions significantly influenced the low molecular weight nitrogen profile and antioxidant activities. Inoculation with the starter culture P200S34 increased free amino acids and amines, such as methionine and tyramine, but other nitrogen compounds also increased the antioxidant activity of the low molecular weight nitrogen extracts. Thus, these starter cultures in Iberian sausages can contribute to delaying oxidative changes during storage.     

Comparison of the effects of a commercial and an autochthonous Pediococcus acidilactici and Staphylococcus vitulus starter culture on the sensory and safety properties of a traditional Iberian dry‐fermented sausage “salchichón”

The effect of the addition of an autochthonous starter culture and commercial starter culture on the physico‐chemical and sensory characteristics of the dry‐fermented sausage ‘salchichon’ was investigated. Sausages were prepared with a mixture of Pediococcus acidilactici MC184 and Staphylococcus vitulus RS34 as the autochthonous starter culture (P184S34), ripened for 86 days, and compared with a commercial starter batch (CS1). Strains of acid lactic bacteria and Staphylococcus were identified in the samples along the ripening process which demonstrated that the inoculation of sausages assures their implantation during the ripening process. However, P184S34 starter culture showed a higher inhibitory effect on enterobacteria and coliform flora than the CS1 batches, guaranteeing a better quality hygienic sausages. Dry‐fermented sausages ripened with P184S34 showed higher amounts of NPN and volatile compounds derived from amino acid catabolism than the CS1 batches. Especially important was the result found in biogenic amines, since the P184S34 reduced their accumulation compared to the commercial starter batch. The inoculation of a decarboxylase‐negative autochthonous starter culture P184S34 reduced the biogenic amine accumulation guaranteeing safety and homogeneity of the products without producing a negative effect on the sensory characteristics of the traditional fermented sausages.     

Impact of pre-selected autochthonous starter cultures on the flavor quality of Iberian dry-fermented "salchichón" sausage with different ripening processes

The purpose of this study was to investigate the impact on flavor quality of 2 autochthonous starter cultures in the manufacture of Iberian dry-fermented "sálchichón." A total of 2 strains of Pediococcus acidilactici (MS200 and MS198) were combined individually with a strain of Staphylococcus vitulus (RS34) to obtain 2 starter cultures: P200S34 and P198S34. The ability of both starter cultures to implant during 2 different manufacturing procedures of "salchichón" was evaluated by 16S rRNA gene sequence analysis. Changes due to starter culture inoculations on volatile compounds were determined by gas chromatography/mass spectrometry, and the impact on flavor quality evaluated by sensorial analysis. The implantation of starter cultures was adequate and did not significantly modify the flavor of the traditional Iberian dry-fermented "salchichón," while modulating the volatile compound profile with respect to that found in sausages with wild microbial populations. The influence of the starter cultures studied was more evident in the shorter sausage processing time. P198S34 was related with the increase in some volatile compounds deriving from lipid degradation. P200S34 contributed to the formation of several volatile compounds arising from lipid degradation, amino acid catabolism, and microbial esterase activity. PRACTICAL APPLICATION: P200S34 and P198S34 are autochthonous starter cultures for specific use in the processing of Iberian dry-fermented sausages. The application of these starters allows obtaining Iberian dry-fermented sausages with sensorial characteristics standardized. It is greatly interesting for processing and marketing industry of these high-quality meat products.     

Role of an autochthonous starter culture and the protease EPg222 on the sensory and safety properties of a traditional Iberian dry-fermented sausage "salchichón"

The effect of the addition of an autochthonous starter culture and the protease EPg222 on the physico-chemical and sensory characteristics of dry-fermented sausage ‘‘salchichon” was investigated. Sausages were prepared with purified EPg222 and Pediococcus acidilactici MS200 and Staphylococcus vitulus RS34 as starter culture (P200S34), separately and together, ripened for 90 days, and compared with a control batch. Dry-fermented sausages ripened with EPg222 and starter culture showed higher amounts of AN and volatile compounds derived from amino acid catabolism than the control, especially in samples in which was added the association of enzyme and starter culture (P200S34 + EPg222). There were clear differences shown by the texture analysis, with the P200S34 + EPg222 batch being less hard. Especially important was the result found in biogenic amines, since the association P200S34 + EPg222 reduced their accumulation compared to the EPg222 batch. The use of EPg222 may be of great interest to improve the sensory characteristics of dry-fermented sausages, but its association with the selected starter culture with low decarboxylase activity is necessary to guarantee healthiness and homogeneity.     

Effect of autochthonous starter cultures isolated from Siahmazgi cheese on physicochemical,microbiological and volatile compound profiles and sensorial attributes of sucuk,a Turkish dry-fermented sausage

The effect of adding autochthonous starter cultures isolated from Siahmazgi cheese, on the physicochemical parameters and microbial counts of sucuk was investigated during the ripening period. SPME–GC/MS was used in volatile compound analysis and a trained group of panelists carried out sensory analysis of the final product. After preliminary screening, three strains of Lactobacillus plantarum, which possess desirable technological properties, were used to prepare three starter cultures: LBP7, LBP10 and LBP14. The addition of LBP7 and LBP14 starter cultures had a significant effect (P < 0.05) on lightness, leading to higher L values compared to control sausages during the ripening period. Both LBP7 and LBP14 sausages showed higher counts of lactic acid bacteria, lower growth of Enterobacteriaceae and Gram-positive catalase-positive cocci and greatly lowered the pH value compared to control sausages throughout the ripening process. At the end of the ripening process, lactic acid bacteria counts were affected (P < 0.05) by the addition of starter culture since higher counts were observed in sausages prepared with LBP7 (9.14 log CFU/g) and LBP14 (8.96 log CFU/g) batches. The decrease of water activity during the ripening of sausages was not affected by the various starters. The texture profiles of all sausages were similar except for LBP10, which showed lower hardness and gumminess during ripening. Under the conditions of the study, volatile compounds were mainly from spices, and no marked differences were found among inoculated sausages. However, sensory evaluation revealed that most of the sensory attributes were scored higher for inoculated sausages than for the control ones. Therefore, LBP7 and LBP14 could be promising candidates for inclusion as starter cultures for the manufacture of sucuk.     

Use of molecular methods to characterize the bacterial community and to monitor different native starter cultures throughout the ripening of Galician chorizo

The development of Lactobacillus and Staphylococcus strains used as starter cultures throughout the ripening of Galician chorizo, a traditional dry fermented sausage from the north-west of Spain, was monitored combining different molecular-based techniques. The bacterial diversity occurring in the inoculated sausages at the beginning and the end of the ripening was also studied and compared to the indigenous population in an uninoculated control batch.     

Effect of selected strains of Debaryomyces hansenii on the volatile compound production of dry fermented sausage “salchichón”

Different biotypes of Debaryomyces hansenii, characterized by mitochondrial DNA (mtDNA) restriction analysis, were inoculated in dry fermented sausages to evaluate their influence as single starter culture on volatile compound generation throughout the ripening process. Similar evolution of physicochemical parameters and microbial population was observed in both uninoculated and inoculated sausages. The tested biotypes modified the volatile compound profile of sausages specially in esters, branched alcohols and aldehydes. The biotype of D. hansenii with the E mtDNA restriction pattern is the most suitable to be used as starter culture since it produced volatile compounds involved in flavour development of dry-cured meat products such as 3-methylbutanol, 3-methylbutanal and 2-propanone. Moreover, the use of D. hansenii strains with the B, C2 and E mtDNA restriction patterns, as a mixed starter culture, should be also considered to generate low amount of sulphur compounds in dry-cured meat products.     

Biochemical and sensory characteristics of traditional fermented sausages of Vallo di Diano (Southern Italy) as affected by the use of starter cultures

In this study, two strains of Staphylococcus xylosus isolated from traditional fermented sausages of Vallo di Diano (Southern Italy) were used in combination with an acidifying strain of Lactobacillus curvatus as starter culture for the production of fermented sausages. Two starter formulation were developed combining the proteolytic but not lipolytic (prt(+), lip(-)) S. xylosus CVS11 with the L. curvatus AVL3 (starter S1) and the S. xylosus FVS21 (prt(-), lip(+)) with the same strain of L. curvatus (starter S2). Proteolysis and lipolysis were observed during ripening by the increase in total free amino acids (FAA) and free fatty acids (FFA), respectively. Such activities were observed in both started and non started sausages (control). Moreover, the proteolytic and lipolytic activities were detected in products started by both formulations irrespective of the presence of such activities in the strains used. Therefore, it was not possible to conclude whether the effect of proteolysis and lipolysis during ripening of the started fermented sausages was due to the activity of the starter cultures or to the action of meat endogenous enzymes.     

Tyrosine- and histidine-decarboxylase positive lactic acid bacteria and enterococci in dry fermented sausages

Lactic acid bacteria (LAB) and enterococci were isolated immediately after stuffing (day 0), at the end of ripening (28th day) and at the end of storage (112th day) from dry fermented sausages produced by two different producers (K; R) in two diameters (4.5 and 7 cm) using either of two spice mixtures (P; H) and either of two starter cultures (Pediococcus pentosaceus, C; Lactobacillus curvatus + Staphylococcus carnosus, F), resulting in a total of 16 different combinations. Tyrosine-decarboxylase DNA sequence (tyrdc) was identified on average in 88% and 44% of enterococci and LAB isolates, respectively at the end of ripening, the corresponding figures regarding histidine-decarboxylase gene sequence (hisdc) was 71% and 16%, respectively. Lactobacillus plantarum, L. brevis and L. casei/paracasei, and Enterococcus faecium and Enterococcus faecalis were identified as tyramine/histamine producers in the sausages.     

Impact of Debaryomyces hansenii strains inoculation on the quality of slow dry-cured fermented sausages

Debaryomyces hansenii strains, M4 and P2, isolated from natural fermented sausages were inoculated in slow fermented sausages to study their effect on processing parameters, microbial population, volatile compound and sensory characteristics. The inoculation of D. hansenii strains, M4 and P2, did not affect the ripening process as no differences in pH and Aw were detected. The dominance of the inoculated yeast strains along the process was followed by RAPDs of M13 minisatellite. The inoculated yeasts, P2 and M4, were recovered at the end of the ripening process although P2 appeared in higher counts than M4. The sausages inoculated with P2 resulted in a decrease in lipid oxidation values (TBARS) and a reduction of lipid-oxidation derived aldehydes in addition to a highest acid compound abundance. M4 inoculated sausages resulted in highest sulphur containing compound abundance. However, no differences in consumer acceptance were detected. Moreover, both yeast strains were responsible for the generation of ethyl methyl-branched ester compounds in the dry-cured sausages.     

The Influence of Debaryomyces hansenii, Candida deformans and Candida zeylanoides on the aroma formation of dry-cured “lacón”

The volatile profile of dry-cured “lacón” that has been inoculated with three different yeasts were determined and compared with a non-inoculated dry-cured “lacón”. Yeasts (Debaryomyces hansenii, Candida deformans and Candida zeylanoides) that were used as starter cultures in the present study were selected among yeasts that were isolated from native dry-cured “lacón” at different stages of ripening process. These starters were spread on dry-cured “lacón” surface in order to test their capacity to contribute on the generation of volatile compounds. A total of forty two volatile compounds were detected by dynamic headspace sampling followed by gas chromatography–mass spectrometry analysis. Significant differences (P < 0.001) on the volatile profiles of different batches were found in comparison with non-inoculated samples, showing the highest total area values for the inoculated ones. Esters were the most abundant chemical family in all batches studied except for C. zeylanoides batch, which showed greater amount of hydrocarbons than esters. The second more abundant family was hydrocarbons for control and C. deformans batches (147.6 and 445.24 × 10⁶ area units, respectively), alcohols for D. hansenii (363.77 × 10⁶ area units) and esters for C. zeylanoides (248.33 × 10⁶ area units). However, the aldehyde compound group in control batch samples was found to be significantly higher than in the inoculated ones (P < 0.001). Among inoculated batches, D. hansenii batch showed the lowest hexanal content (14.42 × 10⁶ area units) in comparison with non-inoculated batch (105.99 × 10⁶ area units). Among all batches studied, D. hansenii batch presented the highest area values for esters, alcohols, linear hydrocarbons, ketones, acids and furans; control batch for aldehydes and C. zeylanoides batch for branched hydrocarbons. Therefore, the study showed that every yeast strain produced a specific volatile profile which was also different from that of the control dry-cured “lacón”.     

Influence of starters on chemical, biochemical, and sensory changes in Turkish White-brined cheese during ripening

Turkish White-brined cheese was manufactured using Lactococcus strains (Lactococcus lactis ssp. lactis NCDO763 plus L. lactis ssp. cremoris SK11 and L. lactis ssp. lactis UC317 plus L. lactis ssp. cremoris HP) or without a starter culture, and ripened for 90 d. It was found that the use of starters significantly influenced the physical, chemical, biochemical, and sensory properties of the cheeses. Chemical composition, pH, and sensory properties of cheeses made with starter were not affected by the different starter bacteria. The levels of soluble nitrogen fractions and urea-PAGE of the pH 4.6-insoluble fractions were found to be significantly different at various stages of ripening. Urea-PAGE patterns of the pH 4.6-insoluble fractions of the cheeses showed that considerable degradation of α_s1-casein occurred and that β-casein was more resistant to hydrolysis. The use of a starter culture significantly influenced the levels of 12% trichloroacetic acid-soluble nitrogen, 5% phosphotungstic acid-soluble nitrogen, free amino acids, total free fatty acids, and the peptide profiles (reverse phase-HPLC) of 70% (vol/vol) ethanol-soluble and insoluble fractions of the pH 4.6-soluble fraction of the cheeses. The levels of peptides in the cheeses increased during the ripening period. Principal component and hierarchical cluster analyses of electrophoretic and chromatographic results indicated that the cheeses were significantly different in terms of their peptide profiles and they were grouped based on the use and type of starter and stage of ripening. Levels of free amino acid in the cheeses differed; Leu, Glu, Phe, Lys, and Val were the most abundant amino acids. Nitrogen fractions, total free amino acids, total free fatty acids, and the levels of peptides resolved by reverse phase-HPLC increased during ripening. No significant differences were found between the sensory properties of cheeses made using a starter, but the cheese made without starter received lower scores than the cheeses made using a starter. It was found that the cheese made with strains NCDO763 plus SK11 had the best quality during ripening. It was concluded that the use of different starter bacteria caused significant differences in the quality of the cheese, and that each starter culture contributed to proteolysis to a different degree.     

Effect of selected dairy starter cultures on microbiological, chemical and sensory characteristics of swine and venison (Dama dama) nitrite-free dry-cured sausages

The aim of this study was the evaluation of selected lactic acid bacteria (LAB) starter culture of dairy origin in the production of nitrite-free low-acid fermented venison (Dama dama) sausage (salame di daino) produced in a small-scale plant in Umbria (Italy), and their effect on microbiological, physico-chemical and sensorial properties of the products. Salame di daino was obtained with two different processes: with and without the addition of selected LAB starter cultures. Microbial counts of Enterobacteriaceae, coliform organisms and Pseudomonas spp. were lower in salami made with the addition of starter cultures. Staphylococcus aureus, Salmonella spp, and Listeria monocytogenes after the first week of ripening were only detected from control salami. Control salami were paler and harder, whereas those made with the addition of starter cultures were slightly saltier, juicier and in general more acceptable. Selected dairy-origin starter (SDS) cultures did prevent the growth of both indicators of food safety and of process hygiene and increased the acceptability of full-ripened salami.     

Influence of a defined-strain starter and Lactobacillus plantarum as adjunct culture on volatile compounds and sensory characteristics of Manchego cheese

Three batches of Manchego cheese were manufactured using one of the following starter culture systems: (1) a defined strain starter culture comprising Lactococcus lactis subsp. lactis and Leuconostoc mesenteroides subsp. dextranicum; (2) the above-defined strain starter culture and an adjunct culture (Lactobacillus plantarum), all these strains being isolated from high-quality Manchego cheeses and (3) a commercial starter consisting of two strains of Lactococcus lactis. Differences in volatile profile and the sensory characteristics of these cheeses were studied. After 4 months of ripening, the two batches of cheese made with the defined strain starter cultures obtained the highest scores for sensory attributes and for the overall impression. Additionally, Purge & Trap and SDE analysis showed a more complex volatile profile in these cheeses than in those made with the commercial starter. Extending the maturation time to 8 months for cheeses made with the defined starter cultures led to significant higher levels of free fatty acids and ethyl esters in those cheeses made without adjunct culture. However, panelists did not find significant differences among the sensory characteristics of the two cheeses.     

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.     

Application of Kluyveromyces marxianus, Lactobacillus delbrueckii ssp. bulgaricus and L. helveticus for sourdough bread making

The application of Kluyveromyces marxianus (IFO 288), Lactobacillus delbrueckii ssp. bulgaricus (ATCC 11842) and Lactobacillus helveticus (ATCC 15009) as starter cultures for sourdough bread making was examined. Production of lactic and acetic acids, bread rising, volatile composition, shelf-life and organoleptic quality of the sourdough breads were evaluated. The amount of starter culture added to the flour, the dough fermentation temperature and the amount of sourdough used were examined in order to optimise the bread making process. The use of mixed cultures led to higher total titratable acidities and lactic acid concentrations compared to traditionally made breads. Highest acidity (3.41 g lactic acid/kg of bread) and highest resistance to mould spoilage were observed when bread was made using 50% sourdough containing 1% K. marxianus and 4% L. delbrueckii ssp. bulgaricus. The use of these cultures also improved the aroma of sourdough breads, as shown by sensory evaluations and as revealed by GC–MS analysis.     

Diversity of lactic acid bacteria from Hussuwa, a traditional African fermented sorghum food

The diversity of lactic acid bacteria associated with Hussuwa fermentation, a Sudanese fermented sorghum food, was studied using a polyphasic taxonomical approach. Predominant strains could be well characterised based on a combination of phenotypic tests and genotypic methods such as ARDRA, rep-PCR and RAPD-PCR, as well as 16S rRNA gene sequencing of representative strains. Thus, the majority (128 of 220, 58.3%) of strains exhibited phenotypic properties typical of heterofermentative lactobacilli and of these, 100 strains were characterised more closely using the genotyping methods. The majority (97/100) strains could be characterised as Lactobacillus fermentum strains. Seventy-two of 220 strains (32.7%) showed phenotypic properties that are characteristic of pediococci. Of 41 selected strains investigated by genotyping techniques, 38 (92.7%) could be characterised as Pediococcus acidilactici strains, while three (7.3%) could be characterised as Pediococcus pentosaceus strains. The Hussuwa fermentation thus appears to be dominated by L. fermentum strains and P. acidilactici strains. For this reason, we selected representative and predominant strains as potential starter cultures for Hussuwa fermentation. These strains, L. fermentum strains BFE 2442 and BFE 2282 and P. acidilactici strain BFE 2300, were shown on the basis of RAPD-PCR fingerprinting to predominate in a model fermentation when used as starter cultures inoculated at 1 × 10⁶ CFU/g and to lower the pH of the fermentation to below pH 4.0 within 48 h. These cultures should be studied for further development as starter preparations in pilot scale studies in actual field fermentations.     

Effects of Inoculation of Commercial Starter Cultures on the Quality and Histamine Accumulation in Fermented Sausages

To meet the requirements of high‐quality safe products, starter cultures are used to produce fermented sausages. The effects of 3 commercial starter cultures, namely SM‐194, T‐SPX, and SM‐181, on histamine accumulation and quality parameters including microbial quality, pH, water activity, and total volatile base nitrogen, as well as the color and texture properties, were evaluated during the fermentation and ripening of fermented sausages. Although initial counts of Escherichia coli, Enterobacteriaceae, and Pseudomonas were similar in the 4 batches, the growth of these microorganisms was significantly inhibited (P < 0.05) in batches SM‐194, T‐SPX, and SM‐181 throughout the fermentation and ripening period. The counts of E. coli, Enterobacteriaceae, and Pseudomonas increased to maximum levels of 3.89, 4.41, and 5.15 log₁₀ colony forming units/g in the control sausages, respectively. At the end of ripening, the levels of histamine were 8.85, 0.32, 7.82, and 3.18 mg/kg for batches C, SM‐194, T‐SPX, and SM‐181, respectively. The results revealed that commercial starter cultures, particularly starter cultures SM‐194 and SM‐181, made a great contribution to histamine reduction. In addition, batches inoculated with starter cultures showed a stronger acidification and lower level of total volatile base nitrogen than the control sample during production (P < 0.05). In conclusion, it seems that the inoculation of commercial starter cultures, particularly starter cultures SM‐194 and SM‐181, contributes to improving microbial quality, hygienic quality and food safety of fermented sausages.