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Culture-dependent and -independent investigations of bacterial migration into doenjang from its components meju and solar salt.

ABSTRACT: We determined bacterial migration into doenjang from its components, meju and solar salt using culture-based and 16S rRNA gene-based culture-independent techniques (pyrosequencing of total DNA). Pyrosequencing results suggested that the bacterial communities of meju, but not solar salt, significantly affected those of doenjang communities. Culture-based pyrosequencing analysis yielded similar results. These results indicate that most predominant bacterial species in doenjang migrated from meju, not solar salt. We therefore believe that the present study is one of the most comprehensive comparisons of bacterial communities of fermented soybeans using culture-dependent and -independent methods. Furthermore, pyrosequencing of the V3 and V4 regions of bacterial 16S rRNA did not distinguish among Bacillus amyloliquefaciens, B. siamensis, and B. velezensis as well as between Enterococcus faecium and E. hirae.

SUBMITTER: Lee JM

PROVIDER: S-EPMC7553307 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature

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Culture-dependent and -independent investigations of bacterial migration into doenjang from its components meju and solar salt.

Lee Jung-Min JM Heo Sojeong S Kim Yoon-Su YS Lee Jong-Hoon JH Jeong Do-Won DW

PloS one 20201013 10

We determined bacterial migration into doenjang from its components, meju and solar salt using culture-based and 16S rRNA gene-based culture-independent techniques (pyrosequencing of total DNA). Pyrosequencing results suggested that the bacterial communities of meju, but not solar salt, significantly affected those of doenjang communities. Culture-based pyrosequencing analysis yielded similar results. These results indicate that most predominant bacterial species in doenjang migrated from meju, ...[more]

PMID: 33048960

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Project description:In the central plains of North America, the beetle family Silphidae comprised two subfamilies, Silphinae and Nicrophorinae, differentiated by reproductive behaviors. Silphinae, known as carrion beetles, feed on carrion and fly larvae and produce free-living larvae that receive no parental care. Adult Nicrophorinae, known as burying beetles, prepare a vertebrate carcass into a brood ball and provide biparental care to their offspring. Preparation of a brood ball involves coating the carcass in antimicrobial oral and anal secretions. These secretions contain a community of microbes, referred to as the secretion microbiome, which inhibit carcass microbe succession, preventing normal decomposition. Here, the secretion microbiomes of five species of Nicrophorinae and two species of Silphinae, both sampled from Oklahoma, with additional Nicrophorinae from Nebraska, were characterized using culture-independent analyses to understand and decipher factors shaping diversity and community structure. We identify the core secretion microbiome across Silphidae and show that, while the host subfamily, secretion type, and collection locality had no significant effect on the bacterial community alpha diversity, these factors significantly influenced bacterial community structure. Global and local tests of phylogenetic associations identified 14 genera with phylogenetic signals to the host subfamily and species. Description of the bacterial communities present in silphid secretions furthers our understanding of how these beetles interact with microbes for carcass nutrient processing. Future culture-dependent studies from silphid secretions may identify novel antimicrobials and nontoxic compounds that can act as meat preservatives or sources for antimicrobials.IMPORTANCEThe manuscript explores the secretion bacterial community of carrion and burying beetles of the central plains of North America. A core secretion microbiome of 11 genera is identified. The host subfamily, secretion type, and collection locality significantly affects the secretion microbiome. Future culture-dependent studies from silphid secretions may identify novel antimicrobials and nontoxic compounds that can act as meat preservatives or sources for antimicrobials.

Project description:BackgroundChild-care facilities appear to provide daily opportunities for exposure and transmission of bacteria and viruses. However, almost nothing is known about the diversity of microbial contamination in daycare facilities or its public health implications. Recent culture-independent molecular studies of bacterial diversity in indoor environments have revealed an astonishing diversity of microorganisms, including opportunistic pathogens and many uncultured bacteria. In this study, we used culture and culture-independent methods to determine the viability and diversity of bacteria in a child-care center over a six-month period.ResultsWe sampled surface contamination on toys and furniture using sterile cotton swabs in four daycare classrooms. Bacteria were isolated on nutrient and blood agar plates, and 16S rRNA gene sequences were obtained from unique (one of a kind) colony morphologies for species identification. We also extracted DNA directly from nine representative swab samples taken over the course of the study from both toy and furniture surfaces, and used "universal" 16S rRNA gene bacterial primers to create PCR-based clone libraries. The rRNA gene clones were sequenced, and the sequences were compared with related sequences in GenBank and subjected to phylogenetic analyses to determine their evolutionary relationships. Culturing methods identified viable bacteria on all toys and furniture surfaces sampled in the study. Bacillus spp. were the most commonly cultured bacteria, followed by Staphylococcus spp., and Microbacterium spp. Culture-independent methods based on 16S rRNA gene sequencing, on the other hand, revealed an entirely new dimension of microbial diversity, including an estimated 190 bacterial species from 15 bacterial divisions. Sequence comparisons and phylogenetic analyses determined that the clone libraries were dominated by a diverse set of sequences related to Pseudomonas spp., as well as uncultured bacteria originally identified on human vaginal epithelium. Other sequences were related to uncultured bacteria from wastewater sludge, and many human-associated bacteria including a number of pathogens and opportunistic pathogens. Our results suggest that the child-care facility provided an excellent habitat for slime-producing Pseudomonads, and that diaper changing contributed significantly to the bacterial contamination.ConclusionThe combination of culture and culture-independent methods provided powerful means for determining both viability and diversity of bacteria in child-care facilities. Our results provided insight into the source of contamination and suggested ways in which sanitation might be improved. Although our study identified a remarkable array of microbial diversity present in a single daycare, it also revealed just how little we comprehend the true extent of microbial diversity in daycare centers or other indoor environments.

Dataset Information

Culture-dependent and -independent investigations of bacterial migration into doenjang from its components meju and solar salt.

Publications

Culture-dependent and -independent investigations of bacterial migration into doenjang from its components meju and solar salt.

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