Microbial Diversity Analysis of Fermented Mung Beans (Lu-Doh-Huang) by Using Pyrosequencing and Culture Methods

Abstract

In Taiwanese alternative medicine Lu-doh-huang (also called Pracparatum mungo), mung beans are mixed with various herbal medicines and undergo a 4-stage process of anaerobic fermentation. Here we used high-throughput sequencing of the 16S rRNA gene to profile the bacterial community structure of Lu-doh-huang samples. Pyrosequencing of samples obtained at 7 points during fermentation revealed 9 phyla, 264 genera, and 586 species of bacteria. While mung beans were inside bamboo sections (stages 1 and 2 of the fermentation process), family Lactobacillaceae and genus Lactobacillus emerged in highest abundance; Lactobacillus plantarum was broadly distributed among these samples. During stage 3, the bacterial distribution shifted to family Porphyromonadaceae, and Butyricimonas virosa became the predominant microbial component. Thereafter, bacterial counts decreased dramatically, and organisms were too few to be detected during stage 4. In addition, the microbial compositions of the liquids used for soaking bamboo sections were dramatically different: Exiguobacterium mexicanum predominated in the fermented soybean solution whereas B. virosa was predominant in running spring water. Furthermore, our results from pyrosequencing paralleled those we obtained by using the traditional culture method, which targets lactic acid bacteria. In conclusion, the microbial communities during Lu-doh-huang fermentation were markedly diverse, and pyrosequencing revealed a complete picture of the microbial consortium.

Bacterial communities and volatile compounds in Doubanjiang, a Chinese traditional red pepper paste

“…Furthermore, Enterobacteriaceae family was again identified by MiSeq sequencing analysis with different hypervariable region (V4) of the bacterial 16S rRNA gene; these results confirmed that Enterobacteriaceae family was one of the predominant bacterial communities in Doubanjiang which was in accordance with the previous report that bacteria belonging to the Enterobacteriaceae family are susceptible to grow under low pH (about 4Á5) conditions (Priest and Stewart 2006). The amount of Enterobacteriaceae family is thought to be as a useful indicator for food sanitation (Feng et al 2013) and the species of Enterobacteriaceae are frequently found in food fermentation such as soy sauce (Wei et al 2013) and fermented mung beans (Chao et al 2013). In addition, previous study indicated that some strains of these bacteria identified in this study (e.g.…”

“…) and fermented mung beans (Chao et al . ). In addition, previous study indicated that some strains of these bacteria identified in this study (e.g.…”

Analysis of bacterial and fungal communities in Marcha and Thiat, traditionally prepared amylolytic starters of India

“…Introduction of culture-independent methods and it’s applicability in food microbiology 7 , 17 , has been a motivation for few researchers to profile the microbial community structure of some Asian starter cultures using PCR-DGGE, pyrosequencing, etc. which is suggestive to provide more insight into the microbial diversity of ethnic starters 3 , 5 , 18 – 22 . Rapid evolution in next generation sequencing (NGS) technologies has enabled researchers to have increased accuracy, throughput, with reasonably low cost and in relatively short period of time 17 , 23 .…”

Inhibitory effect of Lactobacillus salivarius on Streptococcus mutans biofilm formation

“…The 64 L. salivarius strains were isolated from various sources, including fermented vegetables (five strains), human saliva (49 strains), and breast milk (10 strains). Identification of bacterial species was performed according to 16S rDNA sequencing, as previously described (Chao et al ., , ), and both K35 and K43 were isolated form human saliva. Lactobacilli and S. mutans were respectively cultured in deMan, Rogosa, and Sharpe (MRS) and brain–heart infusion (BHI) media (BD Difco, Franklin Lakes, NJ) at 37°C for 20 h under anaerobic atmospheric conditions (10% CO 2 , 10% H 2 , 80% N 2 ).…”