• Asian-Australasian Journal of Animal Sciences
• /
• v.33 no.11
• /
• pp.1858-1865
• /
• 2020

Objective: The microbiota of dairy cow milk varies with the season, and this accounts in part for the seasonal variation in mastitis-causing bacteria and milk spoilage. The microbiota of the cowshed may be the most important factor because the teats of a dairy cow contact bedding material when the cow is resting. The objectives of the present study were to determine whether the microbiota of the milk and the cowshed vary between seasons, and to elucidate the relationship between the microbiota. Methods: We used 16S rRNA gene amplicon sequencing to investigate the microbiota of milk, feces, bedding, and airborne dust collected at a dairy farm during summer and winter. Results: The seasonal differences in the milk yield and milk composition were marginal. The fecal microbiota was stable across the two seasons. Many bacterial taxa of the bedding and airborne dust microbiota exhibited distinctive seasonal variation. In the milk microbiota, the abundances of Staphylococcaceae, Bacillaceae, Streptococcaceae, Microbacteriaceae, and Micrococcaceae were affected by the seasons; however, only Micrococcaceae had the same seasonal variation pattern as the bedding and airborne dust microbiota. Nevertheless, canonical analysis of principle coordinates revealed a distinctive group comprising the milk, bedding, and airborne dust microbiota. Conclusion: Although the milk microbiota is related to the bedding and airborne dust microbiota, the relationship may not account for the seasonal variation in the milk microbiota. Some major bacterial families stably found in the bedding and airborne dust microbiota, e.g., Staphylococcaceae, Moraxellaceae, Ruminococcaceae, and Bacteroidaceae, may have greater influences than those that varied between seasons.

• The Plant Pathology Journal
• /
• v.31 no.1
• /
• pp.25-32
• /
• 2015

Pseudomonas coronafaciens causes halo blight on oats and is a plant quarantine bacterium in many countries, including the Republic of Korea. Using of the certificated seed is important for control of the disease. Since effective detection method of P. coronafaciens is not available yet, PCR and TaqMan PCR assays for specific detection of P. coronafaciens were developed in this study. PCR primers were designed from the draft genome sequence of P. coronafaciens LMG 5060 which was obtained by the next-generation sequencing in this study. The PCR primer set Pc-12-F/Pc-12-R specifically amplified 498 bp from the 13 strains of P. coronafaciens isolated in the seven different countries (Canada, Japan, United Kingdom, Zimbabwe, Kenya, Germany, and New Zealand) and the nested primer set Pc-12-ne-F/Pc-12-ne-R specifically amplified 298 bp from those strains. The target-size PCR product was not amplified from the non-target bacteria with the PCR and nested primer sets. TaqMan PCR with Pc-12-ne-F/Pc-12-ne-R and a TaqMan probe, Pc-taqman, which were designed inside of the nested PCR amplicon, generated Ct values which in a dose-dependent manner to the amount of the target DNA and the Ct values of all the P. coronafaciens strains were above the threshold Ct value for positive detection. The TaqMan PCR generated positive Ct values from the seed extracts of the artificially inoculated oat seeds above 10 cfu/ml inoculation level. PCR and TaqMan PCR assays developed in this study will be useful tools to detect and identify the plant quarantine pathogen, P. coronafaciens.

• Journal of Applied Biological Chemistry
• /
• v.59 no.4
• /
• pp.299-304
• /
• 2016

When used for amplicon sequencing, Illumina platforms produce more than hundreds of sequence artefacts, which affects operational taxonomic units based analyses such as differential abundance and network analyses. Nevertheless it has become a major tool for fecal microbial community analysis. In addition, results from sequence-based fecal microbial community analysis vary depending on conditions of samples (i.e., freshness, time of storage and quantity). We investigated if freeze-drying samples could improve quality of sequence data. Our results showed reduced number of possible artefacts while maintaining overall microbial community structure. Therefore, freeze-drying feces prior to DNA extraction is recommended for Illumina-based microbial community analysis.

• Microbiology and Biotechnology Letters
• /
• v.49 no.2
• /
• pp.192-200
• /
• 2021

Kunun-zaki, produced by submerged fermentation of a combination of millet and sorghum, is a popular beverage in Northern Nigeria. Owing to the nature of the process involved in its production, kunun-zaki is highly susceptible to contamination by food spoilage microorganisms, leading to inconsistent quality and short shelf-life. In this study, we investigated various food spices, including cinnamon, garlic, and nutmeg, as potential preservatives that could be used to extend kunun-zaki shelf-life. Kunun-zaki varieties were fermented with each of these spices mentioned above and subjected to bacterial, nutritional, sensory, and quality maintenance assessments (using a twelve-member sensory panel to evaluate the organoleptic properties of kunun-zaki). Bacterial counts in the final products ranged between 10^5-7 CFU/ml. We identified two bacterial genera, Weissella and Enterococcus, based on partial 16S rRNA gene amplicon sequencing. Three amino acids, namely leucine, aspartate, and glutamate, were abundant in all kunun-zaki varieties, while the total essential amino acid content was above 39%, suggesting that kunun-zaki could potentially be considered as a protein-rich food source both for infants and adults. The kunun-zaki products were also rich in carbohydrates, crude proteins, ash, crude fiber, and fat, with contents estimated as 81-84, 8-11, 0.8-4.0, 2.9-3.58, and 5.1-6.3%, respectively. However, this nutritional content depreciated rapidly after 24 h of storage, except for kunun-zaki fermented with garlic, which its crude protein and fat content was maintained for up to 48 h. Our results revealed that organic spices increased the nutritional content of the kunun-zaki varieties and could be potentially be used as natural preservatives for enhancing the kunun-zaki shelf-life. However, garlic might be considered a better alternative based on our preliminary investigation. The presence of the isolated microorganisms in the analyzed kunun-zaki samples should be highlighted to raise awareness on the possible health hazards that could arise from poor handling and processing techniques.

• Asian-Australasian Journal of Animal Sciences
• /
• v.32 no.6
• /
• pp.891-895
• /
• 2019

Objective: Aim of present study was the set up of a fast and reliable protocol using species-specific markers for the quali-quantitative analysis of DNA and the detection of ruminant biological components in dairy products. For this purpose, the promoter of the gene coding for the ${\alpha}$-lactoalbumin (LALBA) was chosen as possible candidate for the presence of short interspersed nuclear elements (SINEs). Methods: DNA was isolated from somatic cells of 120 individual milk samples of cattle (30), Mediterranean river buffalo (30), goat (30), and sheep (30) and the gene promoter region (about 600/700 bp) of LALBA (from about 600 bp upstream of exon 1) has been sequenced. For the development of a single polymerase chain reaction (PCR) protocol that allows the simultaneous identification of DNA from the four species of ruminants, the following internal primers pair were used: 5'-CACTGATCTTAAAGCTCAGGTT-3' (forward) and 5'-TCAGA GTAGGCCACAGAAG-3' (reverse). Results: Sequencing results of LALBA gene promoter region confirmed the presence of SINEs as monomorphic "within" and variable in size "among" the selected species. Amplicon lengths were 582 bp in cattle, 592 bp in buffalo, 655 in goat and 729 bp in sheep. PCR specificity was demonstrated by the detection of trace amounts of species-specific DNA from mixed sources ($0.25ng/{\mu}L$). Conclusion: We developed a rapid PCR protocol for the quali-quantitative analysis of DNA and the traceability of dairy products using a species-specific marker with only one pair of primers. Our results validate the proposed technique as a suitable tool for a simple and inexpensive (economic) detection of animal origin components in foodstuffs.

• Journal of Ginseng Research
• /
• v.45 no.2
• /
• pp.334-343
• /
• 2021

Background: Gut microbiota mainly function in the biotransformation of primary ginsenosides into bioactive metabolites. Herein, we investigated the effects of three prebiotic fibers by targeting gut microbiota on the metabolism of ginsenoside Rb1 in vivo. Methods: Sprague Dawley rats were administered with ginsenoside Rb1 after a two-week prebiotic intervention of fructooligosaccharide, galactooligosaccharide, and fibersol-2, respectively. Pharmacokinetic analysis of ginsenoside Rb1 and its metabolites was performed, whilst the microbial composition and metabolic function of gut microbiota were examined by 16S rRNA gene amplicon and metagenomic shotgun sequencing. Results: The results showed that peak plasma concentration and area under concentration time curve of ginsenoside Rb1 and its intermediate metabolites, ginsenoside Rd, F2, and compound K (CK), in the prebiotic intervention groups were increased at various degrees compared with those in the control group. Gut microbiota dramatically responded to the prebiotic treatment at both taxonomical and functional levels. The abundance of Prevotella, which possesses potential function to hydrolyze ginsenoside Rb1 into CK, was significantly elevated in the three prebiotic groups (P < 0.05). The gut metagenomic analysis also revealed the functional gene enrichment for terpenoid/polyketide metabolism, glycolysis, gluconeogenesis, propanoate metabolism, etc. Conclusion: These findings imply that prebiotics may selectively promote the proliferation of certain bacterial stains with glycoside hydrolysis capacity, thereby, subsequently improving the biotransformation and bioavailability of primary ginsenosides in vivo.

• The Plant Pathology Journal
• /
• v.38 no.4
• /
• pp.313-322
• /
• 2022

Seed-borne pathogens in crops reduce the seed germination rate and hamper seedling growth, leading to significant yield loss. Due to the growing concerns about environmental damage and the development of resistance to agrochemicals among pathogen populations, there is a strong demand for eco-friendly alternatives to synthetic chemicals in agriculture. It has been well established during the last few decades that plant seeds harbor diverse microbes, some of which are vertically transmitted and important for plant health and productivity. In this study, we isolated culturable endophytic bacteria and fungi from soybean seeds and evaluated their antagonistic activities against common bacterial and fungal seed-borne pathogens of soybean. A total of 87 bacterial isolates and 66 fungal isolates were obtained. Sequencing of 16S rDNA and internal transcribed spacer amplicon showed that these isolates correspond to 30 and 15 different species of bacteria and fungi, respectively. Our antibacterial and antifungal activity assay showed that four fungal species and nine bacterial species have the potential to suppress the growth of at least one seed-borne pathogen tested in the study. Among them, Pseudomonas koreensis appears to have strong antagonistic activities across all the pathogens. Our collection of soybean seed endophytes would be a valuable resource not only for studying biology and ecology of seed endophytes but also for practical deployment of seed endophytes toward crop protection.

• Animal Bioscience
• /
• v.36 no.9
• /
• pp.1453-1464
• /
• 2023

Objective: This study investigated the changes in bacterial communities within decomposing swine microcosms, comparing soil with or without intact microbial communities, and under aerobic and anaerobic conditions. Methods: The experimental microcosms consisted of four conditions: UA, unsterilized soil-aerobic condition; SA, sterilized soil-aerobic condition; UAn, unsterilized soil-anaerobic condition; and San, sterilized soil-anaerobic condition. The microcosms were prepared by mixing 112.5 g of soil and 37.5 g of ground carcass, which were then placed in sterile containers. The carcass-soil mixture was sampled at day 0, 5, 10, 30, and 60 of decomposition, and the bacterial communities that formed during carcass decomposition were assessed using Illumina MiSeq sequencing of the 16S rRNA gene. Results: A total of 1,687 amplicon sequence variants representing 22 phyla and 805 genera were identified in the microcosms. The Chao1 and Shannon diversity indices varied in between microcosms at each period (p<0.05). Metagenomic analysis showed variation in the taxa composition across the burial microcosms during decomposition, with Firmicutes being the dominant phylum, followed by Proteobacteria. At the genus level, Bacillus and Clostridium were the main genera within Firmicutes. Functional prediction revealed that the most abundant Kyoto encyclopedia of genes and genomes metabolic functions were carbohydrate and amino acid metabolisms. Conclusion: This study demonstrated a higher bacteria diversity in UA and UAn microcosms than in SA and SAn microcosms. In addition, the taxonomic composition of the microbial community also exhibited changes, highlighting the impact of soil sterilization and oxygen on carcass decomposition. Furthermore, this study provided insights into the microbial communities associated with decomposing swine carcasses in microcosm.

• Animal Bioscience
• /
• v.36 no.11
• /
• pp.1727-1737
• /
• 2023

Objective: The poultry industry is a primary source of animal protein worldwide. The gut microbiota of poultry birds, such as chickens and ducks, is critical in maintaining their health, growth, and productivity. This study aimed to identify longitudinal changes in the gut microbiota of laying hens from birth to the pre-laying stage. Methods: From a total of 80 Hy-Line Brown laying hens, birds were selected based on weight at equal intervals to collect feces (n = 20 per growth) and ileal contents (n = 10 per growth) for each growth stage (days 10, 21, 58, and 101). The V4 regions of the 16S rRNA gene were amplified after extracting DNA from feces and ileal contents. Amplicon sequencing was performed using Illumina, followed by analysis. Results: Microbial diversity increased with growth stages, regardless of sampling sites. Microbial community analysis indicated that Firmicutes, Proteobacteria, and Bacteroidetes were the dominant phyla in the feces and ileal. The abundance of Lactobacillus was highest on day 10, and that of Escherichia-shigella was higher on day 21 than those at the other stages at the genus level (for the feces and ileal contents; p<0.05). Furthermore, Turicibacter was the most abundant genus after changing feed (for the feces and ileal contents; p<0.05). The fecal Ruminococcus torques and ileal Lysinibacillus were negatively correlated with the body weights of chickens (p<0.05). Conclusion: The gut microbiota of laying hens changes during the four growth stages, and interactions between microbiota and feed may be present. Our findings provide valuable data for understanding the gut microbiota of laying hens at various growth stages and future applied studies.

• Journal of Microbiology and Biotechnology
• /
• v.34 no.4
• /
• pp.863-870
• /
• 2024

Meju, a fermented soybean brick, is a key component in soybean foods like doenjang and ganjang, harboring a variety of microorganisms, including bacteria and fungi. These microorganisms significantly contribute to the nutritional and sensory characteristics of doenjang and ganjang. Amplicon-based next-generation sequencing was applied to investigate how the microbial communities of meju fermented at low and high temperatures differ and how this variation affects the microbial communities of doenjang, a subsequently fermented soybean food. Our metagenomic data showed distinct patterns depending on the fermentation temperature. The microbial abundance in the bacterial community was increased under both temperatures during the fermentation of meju and doenjang. Weissella was the most abundant genus before the fermentation of meju, however, it was replaced by Bacillus at high temperature-fermented meju and lactic acid bacteria such as Weissella and Latilactobacillus at low temperature-fermented meju. Leuconostoc, Logiolactobacillus, and Tetragenococcus gradually took over the dominant role during the fermentation process of doenjang, replacing the previous dominant microorganisms. Mucor was dominant in the fungal community before and after meju fermentation, whereas Debaryomyces was dominant under both temperatures during doenjang fermentation. The dominant fungal genus of doenjang was not affected regardless of the fermentation temperature of meju. Strong correlations were shown for specific bacteria and fungi linked to specific fermentation temperatures. This study helps our understanding of meju fermentation process at different fermentation temperatures and highlights different bacteria and fungi associated with specific fermentation periods which may influence the nutritional and organoleptic properties of the final fermented soybean foods doenjang.