Microbial Community Dynamics and Metabolome Changes During Spontaneous Fermentation of Northeast Sauerkraut From Different Households.
ABSTRACT: Sauerkraut, one of the most popular traditional fermented vegetable foods in northern China, has been widely consumed for thousands of years. In this study, the physicochemical characteristics, microbial composition and succession, and metabolome profile were elucidated during the fermentation of traditional northeast sauerkraut sampled from different households. The microbial community structure as determined by high-throughput sequencing (HTS) technology demonstrated that Firmicutes and Proteobacteria were the predominant phyla and Weissella was the most abundant genus in all samples. Except for Weissella, higher relative abundance of Clostridium was observed in #1 sauerkraut, Clostridium and Enterobacter in #2 sauerkraut, and Lactobacillus in #3 sauerkraut, respectively. Meanwhile, Principal component analysis (PCA) revealed significant variances in the volatilome profile among different homemade sauerkraut. Acids and lactones were dominant in the #1 sauerkraut. The #2 sauerkraut had significantly higher contents of alcohols, aldehydes, esters, sulfides, and free amino acids (FAAs). In comparison, higher contents of terpenes and nitriles were found in the #3 sauerkraut. Furthermore, the potential correlations between the microbiota and volatilome profile were explored based on Spearman's correlation analysis. Positive correlations were found between Clostridium, Enterobacter, Lactobacillus, Leuconostoc, Weissella and most volatile compounds. Pseudomonas, Chloroplast, Rhizobium, Aureimonas, and Sphingomonas were negatively correlated with volatile compounds in sauerkraut. This study provided a comprehensive picture of the dynamics of microbiota and metabolites profile during the fermentation of different homemade northeast sauerkraut. The elucidation of correlation between microbiota and volatile compounds is helpful for guiding future improvement of the fermentation process and manufacturing high-quality sauerkraut.
Project description:Northeastern Chinese sauerkraut is a well-known traditional fermented vegetable in China. Incomplete identification of the microorganisms' (bacteria in spontaneous fermentation) diversity and accumulation of nitrite make it difficult to normalize the fermentation process and product qualities of northeastern Chinese sauerkraut. Conventional culturing and polymerase chain reaction-denaturing gradient gel electrophoresis methods were combined to describe microbial structure and diversity. Lactobacillus, Leuconostoc, Enterobacter, Accumulibacter, Thermotoga, Pseudomonas, Clostridium, Rahnella and Citrobacter were predominant microorganisms in different fermentation periods. The pH value and nitrite concentration presented a certain relevance to the amount of lactic acid bacteria. Lactobacillus and Leuconostoc had the ability to decrease nitrite by inhibiting nitrate-reducing bacteria such as Enterobacter. Therefore, Northeastern Chinese sauerkraut should not be eaten until 4 weeks of fermentation for the safety and quality of fermented foods. Northeastern Chinese sauerkraut is rich in lactic acid bacteria, which demonstrate its ability as an excellent probiotic for applications in functional foods.
Project description:This study identified the lactic acid bacteria (LAB) biota and the volatilome profile of 28 typical sourdoughs of Irpinia-a large area of the Campania region of Southern Italy where numerous breads are produced, even today, following the ancient procedures of sourdough fermentation and for which information on the microbiological and sensory profile is lacking in literature. For this purpose, microbial quality, LAB biodiversity, chemical, and technological characteristics, as well as aroma profile by solid-phase microextraction technique (SPME)-gas chromatography/mass spectrometry (GC/MS) of Irpinian sourdoughs were investigated. The dominant LAB microbiota was examined by both culture-dependent and culture-independent methods Polymerase Chain Reaction/Denaturing Gradient Gel Electrophoresis (PCR-DGGE). Results showed a high biodiversity in LAB community whereas the most frequent lactobacilli species recognized were Lactobacillus plantarum (ca. 22% of total LAB isolates), Lactobacillus sanfranciscensis (11%), Lactobacillus paralimentarius (8%), and Lactobacillus rossiae (6.5%), whereas LAB cocci could be mainly referred to Pediococcus pentosaceus (9.5% of total LAB isolates), Leuconostoc spp. (7.8%), and Weissella cibaria (7.7%). Sourdoughs were characterized by the dominance of one or two LAB species, thus proving that the environment influences the selection and the establishment of few key LAB species and that no specific correlation can be traced between microbial composition and geographical origin of the samples. Furthermore, although sourdoughs were characterized by different qualitative and quantitative volatile organic compound (VOC) compositions, no noticeable correlation between volatilome profile and geographical origin was found. However, it emerged that for more isolated locations, it was possible to find the existence of microbial biotypes and sensory profiles with a strong identity, thus revealing the existence of highly traditional and evocative bread recipes in those geographical contexts.
Project description:Bee-collected pollen (BCP) is currently receiving increasing attention as a dietary supplement for humans. In order to increase the accessibility of nutrients for intestinal absorption, several biotechnological solutions have been proposed for BCP processing, with fermentation as one of the most attractive. The present study used an integrated metabolomic approach to investigate how the use of starter cultures may affect the volatilome and the profile of bioaccessible phenolics of fermented BCP. BCP fermented with selected microbial starters (Started-BCP) was compared to spontaneously fermented BCP (Unstarted-BCP) and to unprocessed raw BCP (Raw-BCP). Fermentation significantly increased the amount of volatile compounds (VOC) in both Unstarted- and Started-BCP, as well as modifying the relative proportions among the chemical groups. Volatile free fatty acids were the predominant VOC in Unstarted-BCP. Started-BCP was differentiated by the highest levels of esters and alcohols, although volatile free fatty acids were always prevailing. The profile of the VOC was dependent on the type of fermentation, which was attributable to the selected <i>Apilactobacillus kunkeei</i> and <i>Hanseniaspora uvarum</i> strains used as starters, or to the variety of yeasts and bacteria naturally associated to the BCP. Started-BCP and, to a lesser extent, Unstarted-BCP resulted in increased bioaccessible phenolics, which included microbial derivatives of phenolic acids metabolism.
Project description:The comprehensive bacterial populations and metabolites profile in fermented feed is unclear, which may have significant effects on the stability of fermented feed quality and animal gut health. In this study, 16S rRNA gene sequencing and liquid chromatography-mass spectrometry were used to explore the bacterial populations and metabolites profile in the fermented feed incubated with probiotics (MF) or without probiotics (SF). The probiotics were a combination of Lactobacillus salivarius, Bacillus subtilis, and Saccharomyces cerevisiae. The pH and lactic acid levels were higher in MF than in SF (P < 0.05), while the total volatile fatty acid content was lower (P < 0.05). Interestingly, after fermentation, the most abundant bacterial genus in MF was Enterococcus, rather than the added probiotics Lactobacillus or Bacillus. Weissella and a few potential pathogens (Enterobacter, Escherichia-Shigella, and Pantoea) were dominant in SF (P < 0.05). Metabolomics analysis identified 32 different metabolites in the two types of fermented feed. These metabolites enriched in MF, such as maleic acid, phenylacetic acid, ethyl linoleate, dihomo-gamma-linolenic acid, and L-theanine had potential antimicrobial activities. Conclusively, the addition of probiotics enriched a few potentially beneficial microbes and small molecular compounds with antimicrobial activities, and inhibited the potential pathogens in fermented feed.
Project description:Despite recent interest in microbial communities of fermented foods, there has been little inquiry into the bacterial community dynamics of sauerkraut, one of the world’s oldest and most prevalent fermented foods. In this study, we utilize 16S rRNA amplicon sequencing to profile the microbial community of naturally fermented sauerkraut throughout the fermentation process while also analyzing the bacterial communities of the starting ingredients and the production environment. Our results indicate that the sauerkraut microbiome is rapidly established after fermentation begins and that the community is stable through fermentation and packaging for commercial sale. Our high-throughput analysis is in agreement with previous studies that utilized traditional microbiological assessments but expands the identified taxonomy. Additionally, we find that the microbial communities of the starting ingredients and the production facility environment exhibit low relative abundance of the lactic acid bacteria that dominate fermented sauerkraut.
Project description:Previous studies using traditional biochemical identification methods to study the ecology of commercial sauerkraut fermentations revealed that four species of lactic acid bacteria, Leuconostoc mesenteroides, Lactobacillus plantarum, Pediococcus pentosaceus, and Lactobacillus brevis, were the primary microorganisms in these fermentations. In this study, 686 isolates were collected from four commercial fermentations and analyzed by DNA fingerprinting. The results indicate that the species of lactic acid bacteria present in sauerkraut fermentations are more diverse than previously reported and include Leuconostoc citreum, Leuconostoc argentinum, Lactobacillus paraplantarum, Lactobacillus coryniformis, and Weissella sp. The newly identified species Leuconostoc fallax was also found. Unexpectedly, only two isolates of P. pentosaceus and 15 isolates of L. brevis were recovered during this study. A better understanding of the microbiota may aid in the development of low-salt fermentations, which may have altered microflora and altered sensory characteristics.
Project description:Lactic acid fermentation is one of the oldest methods used worldwide to preserve cereals and vegetables. Europe and Asia have long and huge traditions in the manufacturing of lactic acid bacteria (LAB)-fermented foods. They have different cultures, religions and ethnicities with the available resources that strongly influence their food habits. Many differences and similarities exist with respect to raw substrates, products and microbes involved in the manufacture of fermented products. Many of them are produced on industrial scale with starter cultures, while others rely on spontaneous fermentation, produced homemade or in traditional events. In Europe, common LAB-fermented products made from cereals include traditional breads, leavened sweet doughs, and low and non-alcoholic cereal-based beverages, whereas among vegetable ones prevail sauerkraut, cucumber pickles and olives. In Asia, the prevailing LAB-fermented cereals include acid-leavened steamed breads or pancakes from rice and wheat, whereas LAB-fermented vegetables are more multifarious, such as kimchi, sinki, khalpi, dakguadong, jiang-gua, soidon and sauerkraut. Here, an overview of the main Euro-Asiatic LAB-fermented cereals and vegetables was proposed, underlining the relevance of fermentation as a tool for improving cereals and vegetables, and highlighting some differences and similarities among the Euro-Asiatic products. The study culminated in "omics"-based and future-oriented studies of the fermented products.
Project description:Xiaoqu is a traditional fermentation starter that is used for Chinese liquor production. Although microorganisms in the starters are closely associated with the quality and flavor of liquor, knowledge of the microbiota in xiaoqu is still far from complete, let alone the starters produced by new processes. Here, Illumina MiSeq high-throughput sequencing was applied to study bacterial composition in three types of xiaoqu used in Cantonese soybean-flavor (Chi-flavor) liquor, namely two traditional starters (Jiu Bing and Bing Wan) and a Round-Koji-maker starter (San qu) produced by the automatic starter-making disk machine. The results showed bacterial diversity in traditional starters was similar and higher than that in the Round-Koji-maker starter. Lactobacillus and Pediococcus were the dominant genera in all starters, while other different dominant genera also existed in different starters, which were Weissella, Acetobacter, and Gluconobacter for Jiu Bing, Weissella for Bing Wan, and Bacillus, Acetobacter, Acinetobacter and Klebsiella for San qu, respectively. Meanwhile, Cytophagaceae, one particular microbial family, and some pathogens including Klebsiella, Cronobacter, and Enterobacter were also found in San qu, indicating the automatic starter-making disk machine should be ameliorated before applied into industrial production. These results enriched our knowledge on xiaoqu-related microorganisms and might be helpful in industrial Chi-flavor liquor production and the development of fermentation technology.
Project description:Changes in the microbial gene content and abundance can be analyzed to detect shifts in the microbiota composition due to the use of a starter culture in the food fermentation process, with the consequent shift of key metabolic pathways directly connected with product acceptance. Meat fermentation is a complex process involving microbes that metabolize the main components in meat. The breakdown of carbohydrates, proteins, and lipids can lead to the formation of volatile organic compounds (VOCs) that can drastically affect the organoleptic characteristics of the final products. The present meta-analysis, performed with the shotgun DNA metagenomic approach, focuses on studying the microbiota and its gene content in an Italian fermented sausage produced by using a commercial starter culture (a mix of Lactobacillus sakei and Staphylococcus xylosus), with the aim to discover the connections between the microbiota, microbiome, and the release of volatile metabolites during ripening. The inoculated fermentation with the starter culture limited the development of Enterobacteriaceae and reduced the microbial diversity compared to that from spontaneous fermentation. KEGG database genes associated with the reduction of acetaldehyde to ethanol (EC 126.96.36.199), acetyl phosphate to acetate (EC 188.8.131.52), and 2,3-butanediol to acetoin (EC 184.108.40.206) were most abundant in inoculated samples (I) compared to those in spontaneous fermentation samples (S). The volatilome profiles were highly consistent with the abundance of the genes; elevated acetic acid (1,173.85 ?g/kg), ethyl acetate (251.58 ?g/kg), and acetoin (1,100.19 ?g/kg) were observed in the presence of the starters at the end of fermentation. Significant differences were found in the liking of samples based on flavor and odor, suggesting a higher preference by consumers for the spontaneous fermentation samples. Inoculated samples exhibited the lowest scores for the liking data, which were clearly associated with the highest concentration of acetic acid.IMPORTANCE We present an advance in the understanding of meat fermentation by coupling DNA sequencing metagenomics and metabolomics approaches to describe the microbial function during this process. Very few studies using this global approach have been dedicated to food, and none have examined sausage fermentation, underlying the originality of the study. The starter culture drastically affected the organoleptic properties of the products. This finding underlines the importance of starter culture selection that takes into consideration the functional characteristics of the microorganism to optimize production efficiency and product quality.
Project description:The microbial community diversity and succession of Chinese Sichuan sausages during the spontaneous fermentation were demonstrated using high-throughput sequencing technology. The bacterial diversity was abundant and the succession of bacterial community along the direction of Lactobacillus spp. increased and Weissella spp. decreased. While fungal diversity was single and trace fungal population was detected. The core functional microbiota were lactic acid bacteria, including Lactobacillus spp., Weissella spp. and Pediococcus spp. In initial fermentation, Weissella spp. was the dominant bacteria and its relative abundance was 49.84%, but then its relative abundance decreased to 11.96% during fermentation before recovering to 26.74% at the end of fermentation. Meanwhile, Lactobacillus spp. rose from 24.70 to 55.74% and became the dominant genus. Moreover, Pediococcus spp. increased from 0.06 to 18.05% on day 20 but then decreased to 1.89% on day 30. These results revealed that the primary microorganisms contributing to spontaneous fermentation of Chinese Sichuan sausages were bacteria, while eukaryotic microorganisms such as yeast scarcely contributed to fermentation.