Genotypic diversity and antagonistic activities of enterococci isolated from pastirma.

The biodiversity of enterococci from pastirma (a traditional Turkish dry-cured meat product) by genotypic identification and the antagonistic activities of strains were investigated. Pastirma samples taken from 20 different small-scale factories were subjected to microbiological and physicochemical analysis. A hundred enterococci isolates were identified by 16S rRNA gene sequence analysis. To determine antagonistic activity of strains, Listeria monocytogenes and Staphylococcus aureus were used. The lactic acid bacteria and Micrococcus/Staphylococcus counts were ≥ 6 log cfu/g in 55% of the samples and 75% of the samples, respectively. Enterobacteriaceae was generally below the detectable level (< 2 log cfu/g). The enterococci count was higher than 6 log cfu/g in 30-35% of the samples, depending on the medium used. The enterococci isolates (100 isolates) were identified as E. faecium (80 strains), E. faecalis (19 strains) and E. hirae (1 strain) in genotypic identification. The nine E. faecium strains showed antagonistic activity against L. monocytogenes in the well diffusion test. In contrast, in the same antagonistic activity test, all of the strains had no antagonistic activity against S. aureus. Further studies could be planned to characterize E. faecium strains that show antagonistic activity against L. monocytogenes.

The Effect of a Chitosan/TiO2-Nanoparticle/ Rosmarinic Acid-Based Nanocomposite Coating on the Preservation of Refrigerated Rainbow Trout Fillets (Oncorhynchus mykiss).

The aim of this study was to determine the effect of chitosan (CH)-based nanocomposite coating applications [chitosan+TiO2 (CHT) and chitosan+TiO2+rosmarinic acid (CHTRA)] on changes in quality attributes of rainbow trout fillets during cold storage (4°C). Fish fillets were randomly divided into four groups and subjected to treatments (CH, CHT, CHTRA, and control). After treatments, the groups were packaged under a modified atmosphere (40% CO2+30% O2+30% N2) and stored at 4°C for 18 days. During cold storage, the samples were subjected to physico-chemical and microbiological analyses. During storage, CH, CHT, and CHTRA treatments showed lower aerobic mesophilic and psychrotrophic bacteria counts than the control. However, the differences between coating treatments were not significant. The highest mean pH value was determined in the control group. As the storage time increased, the thiobarbituric acid reactive substances value increased. At the end of the storage period, no significant differences were observed between the treatments, including in the control group. The total volatile basic nitrogen (TVB-N) level in the control group was above 25 mg/100 g on day 15 of storage. However, the TVB-N level in the treatment groups was below 20 mg/100 g on day 18. It was also determined that coating application×storage period interaction had a significant effect on all color parameters (p<0.01). At the end of storage, the highest CIE L* was observed in CHTRA treatment. However, the value of this treatment did not differ from that of the CH treatment.

Partial Replacement of NaCl by KCl, MgCl2 and CaCl2 Chloride Salts in the Production of Sucuk: Effects on Volatile Compounds, Lipid Oxidation, Microbiological and Sensory Properties.

The effects of different chloride salt mixtures (I-control: 100% NaCl, II: 50:50-NaCl:KCl, III: 50:30:20-NaCl:KCl:MgCl2, IV: 50:30:20-NaCl:KCl:CaCl2, V: 50:30:10:10-NaCl:KCl:MgCl2:CaCl2) on the quality properties of sucuk (a dry fermented beef sausage) during ripening were investigated. Lactic acid bacteria reached 8 log cfu/g in the 3 days of fermentation in all treatments. However, salt mixtures including MgCl2 caused an increase in Micrococcus/Staphylococcus. The control group showed the lowest mean aw value at the end of ripening. The salt mixture with 20% CaCl2 showed the lowest mean pH value of 4.97. The mean TBARS value varied between 6.34 and 6.97 µmol MDA/kg but was not affected by the salt mixtures (p > 0.05). According to the results of PCA, salt mixtures I, II and III had a positive correlation in PC1, and PC1 also separated salt mixtures with CaCl2 (IV and V) from other groups. In addition, a strong positive correlation between the control and III group (50:30:20-NaCl:KCl:MgCl2) for sensory properties was determined by heatmap clustering analysis. In addition, the principal component analysis showed that the control, II, and III groups had a stronger correlation with each other for volatile compounds.

The Effects of Sheep Tail Fat, Fat Level, and Cooking Time on the Formation of Nε-(carboxymethyl)lysine and Volatile Compounds in Beef Meatballs.

This study aimed to determine the effects of fat type (sheep tail fat (STF) and beef fat (BF)), fat levels (10, 20, or 30%), and cooking time (0, 2, 4, and 6 min, dry heat cooking at 180 °C) on the carboxymethyl lysine (CML) content in meatballs. pH, thiobarbituric acid reactive substance (TBARS), and volatile compound analyses were also performed on the samples. The use of STF and the fat level had no significant effect on the pH value. The highest TBARS value was observed with the combination of a 30% fat level and STF. CML was not affected by the fat level. The highest CML content was determined in meatballs with STF at a cooking time of 6 min. In the samples cooked for 2 min, no significant difference was observed between STF and BF in terms of the CML content. STF generally increased the abundance of aldehydes. Aldehydes were also affected by the fat level and cooking time. A PCA provided a good distinction between groups containing STF and BF regardless of the fat level or cooking time. Pentanal, octanal, 2,4-decadienal, hexanal, and heptanal were positively correlated with CML.

Evaluation of Autochthonous Coagulase-Negative Staphylococci as Starter Cultures for the Production of Pastirma.

The aim of the study was to investigate the effects of Staphylococcus xylosus 39, S. equorum 53, or S. vitulinus 75, previously isolated from pastirma, on the quality characteristics of pastirma, a Turkish dry-cured meat product, and to evaluate their potential use as starter cultures. The pastirma production was carried out with a traditional method. The control pastirma groups were manufactured without adding any starter culture. At the end of production, the groups were subjected to microbiological and physico-chemical analyses. The pH was above 5.5, and the aw value was below 0.90 in all groups. The strains used exhibited good adaptation to the pastirma. The S. equorum 53 decreased the thiobarbituric acid reactive substances (TBARS) value in pastirma, while the S. xylosus 39 increased the redness (a*) color value. The autochthonous strains caused a decrease in the palmitic acid (C16:0). However, they had no significant effect on the stearic acid (C18:0) and the oleic acid (C18:1n-9c). A total of 41 volatile compounds were identified in the groups. S. vitulinus 75 increased both benzaldehyde and 2-methyl-3-phenylpropanal levels. In addition, the principal component analysis (PCA) of volatile compounds provided a good separation, and PC1 separated S. xylosus 39 from other groups.

Effect of Black Garlic on Microbiological Properties, Lipid Oxidation, Residual Nitrite, Nitrosamine Formation and Sensory Characteristics in a Semi-Dry Fermented Sausage.

This study was conducted with the aim of determining the effects of different black garlic (BG) levels (1%, 2% and 3%) on quality characteristics of a semi-dry fermented sausage (heat-treated sucuk). In addition, the effect of cooking time (0, 1 or 3 min at 180 °C on a hot plate) on nitrosamine formation was investigated. Fresh garlic (FG, 1%) was evaluated as the control group. BG (2% and 3%) caused a reduction in the count of lactic acid bacteria while leading to an increase in pH. FG1% gave the highest number of Micrococcus/Staphylococcus, as well as aw value. The thiobarbituric acid reactive substance (TBARS) value increased with increasing BG levels. FG (1%) showed the highest residual nitrite amount (p < 0.05). The scores for color, taste and general acceptability were reduced by the use of BG (p < 0.05). No significant difference was observed between the garlic treatments in terms of N-Nitrosodimethylamine (NDMA) and N-Nitrosodiethylamine (NDEA) when no additional cooking was applied. Cooking time was determined to have no significant effect on NDMA in 3% BG. The use of BG caused an increase in N-Nitrosopiperidine (NPIP) (p < 0.05). As for PCA, a closer correlation between NPIP and the groups containing BG was observed, while there was a strong correlation between NDMA and the FG group cooked for 3 min. The use of BG caused an increase in NPIP, but affected NDMA and NDEA depending on the cooking time.

A Review on the Role of Lactic Acid Bacteria in the Formation and Reduction of Volatile Nitrosamines in Fermented Sausages.

Nitrosamines are N-nitroso compounds with carcinogenic, mutagenic and teratogenic properties. These compounds could be found at certain levels in fermented sausages. Fermented sausages are considered to be a suitable environment for nitrosamine formation due to acid formation and reactions such as proteolysis and lipolysis during ripening. However, lactic acid bacteria (spontaneous or starter culture), which constitute the dominant microbiota, contribute significantly to nitrosamine reduction by reducing the amount of residual nitrite through nitrite degradation, and pH decrease has an important effect on the residual nitrite amount as well. These bacteria also play an indirect role in nitrosamine reduction by suppressing the growth of bacteria that form precursors such as biogenic amines. In recent years, research interest has focused on the degradation or metabolization of nitrosamines by lactic acid bacteria. The mechanism by which these effects are seen has not been fully understood yet. In this study, the roles of lactic acid bacteria on nitrosamine formation and their indirect or direct effects on reduction of volatile nitrosamines are discussed.

Citric acid production by a novel autochthonous Candida zeylanoides isolate: optimization of process parameters.

In this study, citric acid (CA) production by autochthonous Candida zeylanoides 7.12 was investigated and optimized. Response surface methodology (RSM) was used for the analysis of simultaneous effects of the chosen factors and 2 experiment designs were applied. In the first experimental design, the effects of initial pH value (5.5, 6.0 and 6.5), fermentation time (4, 5 and 6 days) and initial glucose concentration (125, 150 and 175 g/L) on CA production were investigated. Initial pH value was adjusted periodically with NaOH. Results of the statistical analysis showed that the model was found to be not applicable sufficiently to the chosen data. A second experimental design was employed at the same levels of glucose concentration and fermentation time by disabling the pH factor. pH level was kept at 6.5 with CaCO3. Results of the statistical analysis showed that the fit of the model was good and the lack of fit was not significant (P > 0.05). The highest CA concentration of 11.36 g/L was obtained after 6 days of fermentation with an initial glucose concentration of 125 g/L. The results indicated that initial glucose concentration and fermentation time were important parameters for CA production by C. zeylanoides 7.12 and this strain could be used for future studies.

The occurrence of volatile N-nitrosamines in heat-treated sucuk in relation to pH, aw and residual nitrite.

In this study, the occurrence of volatile nitrosamines were investigated in heat-treated sucuk, a kind of semi-dry fermented sausage. The pH, aw and residual nitrite of the samples were also determined. In addition, a principal component analysis (PCA) was also performed in order to elucidate the relationship between nitrosamine and these variables. Significant differences between brands were found in terms of NDMA (N- Nitrosodimethylamine), NPYR (N-Nitrosopyrolidine) and NPIP (N-Nitrosopiperidine) (p < 0.05). NDMA and NPYR varied from 1.71 to 3.57 µg/kg and 1.65 to 7.29 µg/kg, respectively. Higher levels were found for NPIP (5.19 to 16.40 µg/kg). NDEA (N-Nitrosodiethylamine) and NDBA (N- Nitrosodibutylamine) were not found in any of the heat-treated sucuk samples. The residual nitrite content was under 10 mg/kg in all samples. The aw and pH values varied between 0.913 and 0.940 and between 4.28 and 5.47, respectively. In PC1 explaining 72% of the variance, NDMA and NPYR were placed on the negative side, NPIP on the positive side. Residual nitrite and aw were more effective for NPIP, while pH was an important parameter for NDMA and NPYR.

The effects of curing agents on the proteolysis and lipid oxidation of pastirma produced by the traditional method.

In the study, the effects of nitrate and/or nitrite (150 mg/kg KNO3, 300 mg/kg KNO3, 150 mg/kg NaNO2, and 150 mg/kg KNO3 + 150 mg/kg NaNO2) on proteolytic changes (free amino acid composition, SDS-PAGE) in pastirma were investigated. Pastirma samples were also analyzed in terms of some qualitative (pH, aw, TBARS, residual nitrite, salt) properties. The lowest total free amino acid content (1818.3 mg/ 100 g DM) was observed in the combination of 150 mg/kg KNO3 + 150 mg/kg NaNO2, while the highest content (2847.49 mg/ 100 g DM) was observed in 150 mg/kg KNO3. Although the pastirma groups generally exhibited similar SDS-PAGE profiles, differences were detected at some band intensities. The lowest TBARS value (22.24 µmol MDA/kg) was observed in 150 mg/kg KNO3 + 150 mg/kg NaNO2. As a result, the use of 150 mg/kg KNO3 in the pastirma curing process causes more intense proteolysis.

Is sodium carboxymethyl cellulose (CMC) really completely innocent? It may be triggering obesity.

The toxicity of sodium carboxymethyl cellulose (CMC), which has GRAS status and has been determined as "ADI non specified", was re-evaluated with a new modelling and molecular-based data. For this purpose, CMC, a food additive, was injected to the yolk sac (food) of the zebrafish embryo by the microinjection method at the 4th hour of fertilization at different concentrations. As a result, it was found that CMC showed no toxic effects within the framework of the parameters studied. But, we determined increasing lipid accumulation in zebrafish embryos exposed to CMC in a dose-dependent manner. To elucidate the mechanism underlying this lipid accumulation, the expression levels of genes related to obesity-linked lipid metabolism were examined. Our findings show that while CMC does not cause a toxic effect in zebrafish embryos, it can lead important effects on lipid metabolism by causing changes in the expression of some genes associated with obesity.

Technological properties of autochthonous Lactobacillus plantarum strains isolated from sucuk (Turkish dry-fermented sausage).

PURPOSE: Five Lactobacillus strains isolated from sucuk (Turkish dry-fermented sausage) were studied for their genetic and technological properties. METHODS: For genotypic identification, strains 16S rRNA gene sequences were used. To determine the antimicrobial activity of strains, seven foodborne pathogens were tested. Strains technological properties were characterized. RESULTS: These strains were identified as Lactobacillus plantarum by 16S rRNA gene sequence analysis and the phylogenetic tree obtained by neighbor-joining method allowed grouping of these strains into three subgroups. L. plantarum strains showed antagonistic activities against Staphylococcus aureus, Listeria monocytogenes, Bacillus cereus, and Micrococcus luteus strains. PCR assay, using specific primers, showed the presence of bacteriocin (plantaricin) encoding genes in all L. plantarum strains tested. Antimicrobial metabolite production of these strains started at log phase and reached the maximum level at the end of the stationary phase. Regarding their technological properties, better growth was observed at 25 °C compared with 15 °C and 45 °C. The isolates which grown well within the pH scale pH 4.5-6.5 range additionally showed a decent growth at 6.5% salt concentration. It has been found that strains do not exhibit lipolytic and proteolytic activities nor have lysine, ornithine, and arginine decarboxylase activity. On the other hand, one strain showed weak nitrate reductase activity, and four strains produced acetoin from glucose. In addition, all strains were DL-lactic acid producers. Consequently, L. plantarum strains isolated exhibited some biochemical properties required for a starter culture in sucuk and similar products. CONCLUSIONS: All identified strains may be a protective culture in the production of fermented meat products. In particular, L. plantarum S51 was distinguished from other isolates due to the inability to form acetoin from glucose. Further work will be needed to characterize L. plantarum strains as starter culture.

Nitrosamine formation in a semi-dry fermented sausage: Effects of nitrite, ascorbate and starter culture and role of cooking.

In this study, effects of ingoing nitrite level (0, 50, 100 and 150 mg/kg), use of sodium ascorbate, addition of starter culture (Lactobacillus plantarum GM77 + Staphylococcus xylosus GM92) and cooking level (control, medium, medium well, well done and very well done) on nitrosamine formation in heat-treated sucuk, a type of semi-dry fermented sausage, were investigated. The use of ascorbate had no significant effect on NDMA (N-Nitrosodimethylamine) and NPIP (N-Nitrosopiperidine) contents in the presence of starter culture. A higher NPYR (N- Nitrosopyrrolidine) content was detected in the group with starter culture at 150 mg/kg nitrite level in comparison to the group without starter culture. Cooking level affected all identified nitrosamines very significantly. Ingoing nitrite level × cooking level interaction was only effective on NPIP and advanced cooking levels (well done and very well done) at higher ingoing nitrite levels (100 and 150 mg/kg) resulted in significant increases in NPIP content.

Nitrosamines in sucuk: Effects of black pepper, sodium ascorbate and cooking level.

This study determined effects of different black pepper levels (5, 10 or 15 g kg-1), sodium ascorbate usage and cooking level (raw, medium, medium well and well done) on nitrosamine formation in sucuk (a kind of dry fermented sausage). Sodium ascorbate decreased residual nitrite considerably. Cooking level had a very significant effect on nitrosodimethylamine (NDMA), nitrosopyrrolidine (NPYR) and nitrosopiperidine (NPIP) contents. NPIP content increased in sucuk both with and without ascorbate, as cooking level increased. High black pepper level decreased NPIP content in both raw and well done samples. The use of 15 g kg-1 black pepper level caused an increase in both NDMA and NPYR contents in the absence of ascorbate. In contrast, the combination of ascorbate and 15 g kg-1 black pepper resulted in a decrease in NPYR.

Effects of vacuum and high-oxygen modified atmosphere packaging on physico-chemical and microbiological properties of minced water buffalo meat.

OBJECTIVE: In this study, the effects of vacuum (VP) and high-oxygen modified atmosphere (80% O2+20% CO2) packaging (HiOx-MAP) on physico-chemical and microbiological properties of minced water buffalo meat were investigated. METHODS: After minced meat preparation, samples were packaged under VP and HiOx-MAP and stored at 2°C±0.5°C for 14 days. Samples taken on certain days were subjected to total aerobic mesophilic bacteria, total aerobic psychrotrophic bacteria, lactic acid bacteria, Pseudomonas, Enterobacteriaceae and yeast-mold counts as well as pH, color (L*, a*, and b*) and thiobarbituric acid reactive substances (TBARS) analyses. RESULTS: In minced water buffalo meat packaged under HiOx-MAP, TBARS value exceeded 1 mg malondialdehyde/kg on the 4th day of the storage. In VP samples, TBARS value remained close to initial TBARS value during storage. According to the findings, a* value was determined to be high in the HiOx-MAP samples within initial days of the storage. However, no significant changes in a* value were observed in VP samples during storage. In contrast, the mean value of L* was detected as higher in HiOx-MAP sample than VP samples. The count of psychrotrophic bacteria increased more than that of mesophilic bacteria during storage. The growth of Enterobacteriaceae and Pseudomonas was delayed in both the packaging methods. However, lactic acid bacteria exhibited more growth in VP samples compared to MAP samples. CONCLUSION: The lipid oxidation proceeded faster than expected in minced water buffalo meat packed with HiOx-MAP method. This situation adversely affected the a* value. On the other hand, similar microbiological results were obtained in both packing methods.

The effects of different processing conditions on biogenic amine formation and some qualitative properties in pastirma.

Pastirma, a Turkish dry-cured meat product, was cured at two different temperatures (4 or 10 °C) with two different curing agents (150 mg/kg NaNO2 or 300 mg/kg KNO3). The aim of this research was to determine the effects of these factors on biogenic amine content and other qualitative properties (pH, aw, color, residual nitrite, TBARS, NPN-M, microbiological properties). Residual nitrite was below 10 mg/kg in all samples. Both the curing agent and temperature were found to have a very significant effect on the TBARS value, and the curing agent had a significant effect on the NPN-M content. Curing at 10 °C increased the L* value; the use of nitrate increased the a* value. The use of nitrite had a negative effect on the growth of lactic acid bacteria. Micrococcus/Staphylococcus showed good growth in the presence of nitrate. In all samples, Enterobacteriaceae counts were below detectable levels. Neither temperature nor curing agent had significant effects on the amounts of tryptamine, cadaverine, histamine, tyramine, or spermine. There were very significant effects of temperature on the amount of putrescine and of the curing agent on the amount of spermidine.

Volatile compounds and some physico-chemical properties of pastirma produced with different nitrate levels.

OBJECTIVE: The aim of the study was to evaluate the effects of different nitrate levels (150, 300, 450, and 600 ppm KNO3) on the volatile compounds and some other properties of pastirma. METHODS: Pastirma samples were produced under the controlled condition and analyses of volatile compounds, and thiobarbituric acid reactive substances (TBARS) as an indicator of lipid oxidation, non-protein nitrogenous matter content as an indicator of proteolysis, color and residual nitrite were carried out on the final product. The profile of volatile compounds of pastirma samples was analyzed by gas chromatography/mass spectrometry using a solid phase microextraction. RESULTS: Nitrate level had a significant effect on pH value (p<0.05) and a very significant effect on TBARS value (p<0.01). No significant differences were determined in terms of aw value, non-protein nitrogenous substance content, color and residual nitrite between pastirma groups produced by using different nitrate levels. Nitrate level had a significant (p<0.05) or a very significant (p<0.01) effect on some volatile compounds. It was determined that the amounts and counts of volatile compounds were lower in the 450 and especially 600 ppm nitrate levels than 150 and 300 ppm nitrate levels (p<0.05). While the use of 600 ppm nitrate did not cause an increase in residual nitrite levels, the use of 150 ppm nitrate did not negatively affect the color of pastirma. However, the levels of volatile compounds decreased with an increasing level of nitrate. CONCLUSION: The use of 600 ppm nitrate is not a risk in terms of residual nitrite in pastirma produced under controlled condition, however, this level is not suitable due to decrease in the amount of volatile compounds.

The Effects of Geographic Region, Cultivar and Harvest Year on Fatty Acid Composition of Olive Oil.

In this study, the effect of cultivar, harvest year (2010 and 2011) and the geographic regions (Mediterranean, Aegean, Southeastern Anatolia, Marmara and Black Sea) on fatty acid compositions of olive oil was investigated. The proportions of palmitic acid and linoleic acid, which comprised most of the amount of fatty acids after oleic acid, showed variations according to the region and cultivar. The amount of linoleic acid was found higher in some cultivars than that of palmitic acid. In the Aegean, Marmara and Mediterranean Regions, no significant difference was observed between olive cultivars for oleic and linoleic acids. The highest linoleic and lowest oleic acid content were determined in the Black Sea Region. One cultivar (Butko) showed an oleic acid proportion below 55% in this region. The effect of the harvest year on the fatty acid composition of the samples was also found significant. In oils harvested in the second year, a decrease in the amount of linoleic acid was observed, while the amount of oleic acid increased. The highest oleic acid proportion was found in olive oils produced from harvests in the Aegean and Mediterranean regions.

The effects of different levels of orange fiber and fat on microbiological, physical, chemical and sensorial properties of sucuk.

The effects of orange fiber and fat levels on the physical, chemical and microbiological properties of sucuk (Turkish dry-fermented sausage) were investigated. Different levels of orange fiber (0, 2 and 4%) were added to sucuk batters with different levels of fat (10, 15 and 20%). Sucuk samples were analyzed during ripening. Lactic acid bacteria, Micrococcus/Staphylococcus and pH value were effected very significantly by orange fiber. Orange fiber caused a decrease in residual nitrite levels and an increase in the TBARS values. L(∗) and b(∗) values of sucuk samples increased with the use of fiber. Fiber use also decreased the cooking loss. No statistically significant difference was found between control and 2% fiber groups in terms of texture, color, odor taste and general acceptability scores. While fat levels had significant effects on cooking loss, taste and L(∗) values, ripening time effected all analyzed parameters.

Effects of starter cultures and nitrite levels on formation of biogenic amines in sucuk.

The effects of starter cultures, starter A (Lactobacillus sakei+Staphylococcus carnosus) and starter B (Pediococcus acidilactici+Staphylococcus xylosus+Lactobacillus curvatus), nitrite levels (0, 75, and 150ppm) and ripening period on biogenic amine production were investigated in sucuk (Turkish dry-fermented sausage) production. Changes in biogenic amines, pH, water activity, moisture, non-protein nitrogen, nitrite and counts of lactic acid bacteria, Micrococcus/Staphylococcus, Enterobacteriaceae, and moulds-yeasts were determined. Ripening period had a very significant effect (P<0.01) on the all biogenic amines. Sausages produced by fermentation with starter cultures, as compared to natural fermentation (control), had lower amounts of putrescine, cadaverine and tyramine (P<0.05). Starter cultures A and B were found to have almost the same effect on the formation of biogenic amines. Use of nitrite in sucuk production affected the formation of biogenic amines except for spermidine and spermine (P<0.01). Lactic acid bacteria counts in the control group were lower than that of starter culture groups. Lactic acid bacteria, Micrococcus/Staphylococcus, and moulds-yeasts counts increased during ripening period while levels of species of Enterobacteriaceae decreased, which was also found to be under the detectable level (<100cfug(-1)), by day 3 of ripening.