Effects of a Novel Starter Culture on Quality Improvement and Putrescine, Cadaverine, and Histamine Inhibition of Fermented Shrimp Paste.

Fermented shrimp paste is a popular food in Asian countries. However, biogenic amines (BAs) are a typically associated hazard commonly found during the fermentation of shrimp paste and pose a food-safety danger. In this work, an autochthonic salt-tolerant Tetragenococcus muriaticus TS (T. muriaticus TS) strain was used as a starter culture for grasshopper sub shrimp paste fermentation. It was found that with the starter culture, putrescine, cadaverine, and histamine concentrations were significantly lower (p < 0.05) with a maximal reduction of 19.20%, 14.01%, and 28.62%, respectively. According to high-throughput sequencing data, T. muriaticus TS could change the interactions between species and reduce the abundance of bacterial genera positively associated with BAs, therefore inhibiting the BA accumulation during shrimp paste fermentation. Moreover, the volatile compounds during the fermentation process were also assessed by HS-SPME-GC-MS. With the starter added, the content of pyrazines increased, while the off-odor amines decreased. The odor of the shrimp paste was successfully improved. These results indicate that T. muriaticus TS can be used as an appropriate starter culture for improving the safety and quality of grasshopper sub shrimp paste.

Dynamics of Bacterial Composition and Association with Quality Formation and Biogenic Amines Accumulation during Fish Sauce Spontaneous Fermentation.

High-throughput sequencing and high-pressure liquid chromatography (HPLC) methods were used to investigate the influences of microbial dynamics on the quality and biogenic amine (BA) content during fish sauce fermentation. The homogeneity of total viable bacteria and lactic acid bacteria in fish sauce becomes higher as fermentation progresses. Tetragenococcus was the key genus of fish sauce fermentation. Carnobacterium (38.43%) and Lentibacillus (41.01%) were the dominant genera in the samples fermented for 3 months and 18 months, respectively. These three bacterial genera were significantly related to the physicochemical characteristics and characteristic flavors of the sauces. Tetragenococcus was significantly positively correlated with nitrogen oxides, the main characteristic flavor components in fish sauce. The BA content in fish sauce fermentation increased from 106.88 to 376.03 mg/kg, and the content of histamine reached 115.30 mg/kg at the end of fermentation, indicating that fish sauce has health risks. About 66.67% of Lentibacillus isolates were able to produce a large amount of BA, suggesting that Lentibacillus was the key genus for BA accumulation in fish sauce fermentation. Research on reducing the content of BA in fish sauce by intervening with regard to the fermentation temperature showed that a safe fish sauce product could be obtained at the fermentation temperature of about 25°C. These results help us to understand the contribution of microbial community composition to fish sauce fermentation and provide a basis for improving the quality and safety of fermented fish sauce. IMPORTANCE Traditional fermentation of fish sauce is mainly carried out by complex microbial communities from raw anchovies and processing environments. However, it is still unclear how the environmental microbiota influences the quality and the safety of fish sauce products. Therefore, this study comprehensively explained the influence of microorganisms on the quality and safety of fish sauce during the fermentation process in terms of physicochemical characters, flavors, and BA. Additionally, the accumulation of BA in fish sauce fermentation was controlled by intervening in the fermentation temperature. This finding contributes to a deeper understanding of the role of environmental microbiota during fermentation and provides data support for improving the safety of fish sauce.

Impacts of salt-tolerant Staphylococcus nepalensis 5-5 on bacterial composition and biogenic amines accumulation in fish sauce fermentation.

High levels of biogenic amines (BAs) in fermented food can present a health risk to consumers. Microorganisms that can reduce BAs are widely used in fermented foods. However, the mechanism by which microorganisms reduce BAs in foods has not been explored. In this study, we investigated how Staphylococcus nepalensis 5-5 (S. nepalensis 5-5), which was a BA-degrading strain isolated from fish sauce, could reduce BA accumulation in the fish sauce. High-throughput sequencing and HPLC methods were sequentially used to determine the microbial community structure and BA content in fish sauce with/without S. nepalensis 5-5. The results showed that S. nepalensis 5-5 might be a safe strain that could improve the flavor of fish sauce while still exhibiting good BA degradation ability under a high salt environment. The content of BAs in fish sauce inoculated with S. nepalensis 5-5 was significantly decreased compared with the control fish sauce, achieving maximal reductions of 15.74, 14.18 and 16.65% in putrescine, cadaverine and histamine accumulation, respectively. According to high-throughput sequencing data, S. nepalensis 5-5 reduced the abundance of the genera positively associated with BAs, while increasing the number of bacterial genera negatively correlated with BAs in the sample and changed the correlation between some genera and BAs via species interaction. In addition, analysis of amino acid metabolism showed that S. nepalensis 5-5 might use histidine to produce metabolites other than histamine, thereby reducing the production of BAs. These findings not only explained the mechanisms by which the BA level in fish sauce could be reduced but also provided a potential means to control BA production in the fish sauce during the fermentation stage.

Contribution of Microorganisms to Biogenic Amine Accumulation during Fish Sauce Fermentation and Screening of Novel Starters.

In this study, high-throughput sequencing and culture-dependent and HPLC methods were used to investigate the contribution and regulation of biogenic amines (BAs) by dominant microorganisms during fish sauce fermentation. The results showed that the microbial composition constantly changed with the fermentation of fish sauce. Tetragenococcus (40.65%), Lentibacillus (9.23%), Vagococcus (2.20%), Psychrobacter (1.80%), Pseudomonas (0.98%), Halomonas (0.94%) and Staphylococcus (0.16%) were the dominant microflora in fish sauce. The content of BAs gradually increased as the fermentation progressed. After 12 months of fermentation, the histamine content (55.59 mg/kg) exceeded the toxic dose recommended by the Food and Drug Administration (FDA). Correlation analysis showed that dominant microorganisms have a great contribution to the accumulation of BAs. By analyzing the BA production capacity of dominant isolates, the accumulation of BAs in fish sauce might be promoted by Tetragenococcus and Halomonas. Moreover, four strains with high BA reduction ability were screened out of 44 low BA-producing dominant strains, and their influence on BA accumulation in fermented foods was determined. Results demonstrated that Staphylococcus nepalensis 5-5 and Staphylococcus xylosus JCM 2418 might be the potential starters for BA control. The present study provided a new idea for the control of BAs in fermented foods.

Assessment of the Distribution and Safety of Tetragenococcus muriaticus for Potential Application in the Preparation of Chinese Grasshopper Sub Shrimp Paste.

The bacterial profiles of 63 grasshopper sub shrimp paste samples collected from seven typical regions around the Bohai Sea were investigated by high-throughput sequencing. Tetragenococcus muriaticus was found to be the prevailing species present in all the samples, and the presence of T. muriaticus also weakly correlated with the histamine content in the samples. Six T. muriaticus strains with low biogenic amine (BA)-producing ability and deficient in histamine production were identified and subjected to safety assessment. All six strains displayed weak resistance to fifteen known antibiotics as based on the Enterococcus breakpoint values. None of the strains exhibited hemolytic activity or biofilm formation. All strains exhibited were able to grow on MRS agar containing 21% NaCl and expressed amine oxidase and strain-specific proteases and lipases. Most of the strains exhibited acid production at 18% NaCl. Moreover, three of the strains (designated as SG, TS, and QH) with histamine degradation ability were inoculated into separate shrimp paste samples to determine their effect on BA accumulation. The results indicated that the addition of T. muriaticus to shrimp pastes not only led to a significant reduction of BA content in the pastes but also improved the flavor of the pastes. Consequently, these strains may be used as potential candidates for controlling the content of histamine in fermented foods.

The Impact of Microbial Diversity on Biogenic Amines Formation in Grasshopper Sub Shrimp Paste During the Fermentation.

Biogenic amines (BAs) and microbial diversity are important factors affecting food quality and safety in fermented foods. In this study, the bacterial and fungal diversity in grasshopper sub shrimp paste taken at different fermentation times were comprehensively analyzed, while the pH, colony counts, salinity, total volatile base nitrogen (TVB-N) and BA contents were quantitatively determined. In addition, the correlations among the samples with respect to microbial communities and the different parameters investigated especially BAs were also established. By combining the results of spearman correlation heatmap with the contents of BAs produced by the 102 halotolerant bacteria isolated from the grasshopper sub shrimp paste, six major genera of bacteria (Jeotgalibaca, Jeotgalicoccus, Lysinibacillus, Sporosarcina, Staphylococcus, and Psychrobacter) were found to be positively correlated with BA production level, suggesting that these bacteria might have a strong tendency to produce BAs. Other bacteria such as Lentibacillus, Pseudomonas, and Salinicoccus were considered as poor BA producers. The grasshopper sub shrimp paste was characterized by a relatively high abundance of Tetragenococcus, which was the dominant genus during the fermentation process, and it also produced a relatively high level of BAs but the spearman correlation heatmap revealed a negative correlation between T. muriaticus and BA level. Analysis of the species relevance network in grasshopper sub shrimp explained that the actual production of BAs by a certain strain was closely related to other species present in the complex fermentation system.

Association of cytotoxic T lymphocyte antigen-4 +49A/G polymorphism and cancer risk: An updated meta-analysis.

BACKGROUND: Associations between cytotoxic T lymphocyte antigen-4 (CTLA-4) +49A/G polymorphism and cancer risk are inconclusive. We performed this meta-analysis to derive a more precise estimation of the relationship. METHODS: A comprehensive literature search was performed using electronic databases. Odds ratios (ORs) with 95% confidence intervals (CIs) were used to assess the strength of association. RESULTS: A total of 16,358 cases and 19,737 controls from 46 studies were included. Overall, significant association between CTLA-4 +49A/G polymorphism and cancer risk was observed in all genetic models (G vs. A: OR=0.88, 95%CI=0.83-0.93, PH=0.000; GA vs. AA: OR=0.87, 95%CI=0.79-0.97, PH=0.000; GG vs. AA: OR=0.75, 95%CI= 0.65-0.86, PH=0.000; GG vs. GA+AA: OR=0.84, 95%CI=0.79-0.91, PH=0.001; GG+GA vs. AA: OR=0.83, 95%CI=0.74-0.92, PH=0.000). Stratified analysis by cancer type revealed that the CTLA-4+49A/G polymorphism is associated with the decreased risk of cervical cancer, breast cancer, lung cancer, HCC. Further subgroup analysis by ethnicity indicated that there was a statistically decreased cancer risk in Asian population. CONCLUSION: This meta-analysis suggests that CTLA-4+49A/G polymorphism is associated with cancer risk, especially in Asian population.