• 제목/요약/키워드: Illumina MiSeq Sequencing

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Comparison of Fecal Microbial Communities between White and Black Pigs

  • Guevarra, Robin B.;Kim, Jungman;Nguyen, Son G.;Unno, Tatsuya
    • Journal of Applied Biological Chemistry
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    • 제58권4호
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    • pp.369-375
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    • 2015
  • Meat from black pigs (BP) is in high demand compared with that from modern white pig (WP) breeds such as Landrace pigs owing to its high quality. However, the growth rate of black pigs is slower than that of white pig breeds. We investigated differences in the fecal microbial composition between white and black pigs to explore whether these breeds differed in the composition of their gut microbial communities. The swine gut microbiota was investigated using Illumina's MiSeq-based sequencing technology by targeting the V4 region of the 16S rRNA gene. Our results showed that the composition of the gut microbiota was significantly different between the two pig breeds. While the composition of the WP microbiota shifted according to the growth stage, fewer shifts in composition were observed for the BP gut microbiota. In addition, the WP gut microbiota showed a higher Firmicutes/Bacteroidetes ratio compared with that of BP. A high ratio between these phyla was previously reported as an obesity-linked microbiota composition. Moreover, the WP microbiota contained a significantly higher abundance of cellulolytic bacteria, suggesting a possibility of higher fiber digestion efficiency in WP compared to BP. These findings may be important factors affecting growth performance and energy-harvesting capacities in pigs. Our findings of differences in the gut microbiota composition between the two breeds may provide new leads to understand growth rate variation across pig breeds.

Metagenomic Approach to Identifying Foodborne Pathogens on Chinese Cabbage

  • Kim, Daeho;Hong, Sanghyun;Kim, You-Tae;Ryu, Sangryeol;Kim, Hyeun Bum;Lee, Ju-Hoon
    • Journal of Microbiology and Biotechnology
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    • 제28권2호
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    • pp.227-235
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    • 2018
  • Foodborne illness represents a major threat to public health and is frequently attributed to pathogenic microorganisms on fresh produce. Recurrent outbreaks often come from vegetables that are grown close to or within the ground. Therefore, the first step to understanding the public health risk of microorganisms on fresh vegetables is to identify and describe microbial communities. We investigated the phyllospheres on Chinese cabbage (Brassica rapa subsp. pekinensis, N = 54). 16S rRNA gene amplicon sequencing targeting the V5-V6 region of 16S rRNA genes was conducted by employing the Illumina MiSeq system. Sequence quality was assessed, and phylogenetic assessments were performed using the RDP classifier implemented in QIIME with a bootstrap cutoff of 80%. Principal coordinate analysis was performed using a weighted Fast UniFrac matrix. The average number of sequence reads generated per sample was 34,584. At the phylum level, bacterial communities were composed primarily of Proteobacteria and Bacteroidetes. The most abundant genera on Chinese cabbages were Chryseobacterium, Aurantimonadaceae_g, Sphingomonas, and Pseudomonas. Diverse potential pathogens, such as Pantoea, Erwinia, Klebsiella, Yersinia, Bacillus, Staphylococcus, Salmonella, and Clostridium were also detected from the samples. Although further epidemiological studies will be required to determine whether the detected potential pathogens are associated with foodborne illness, our results imply that a metagenomic approach can be used to detect pathogenic bacteria on fresh vegetables.

Universal Indicators for Oil and Gas Prospecting Based on Bacterial Communities Shaped by Light-Hydrocarbon Microseepage in China

  • Deng, Chunping;Yu, Xuejian;Yang, Jinshui;Li, Baozhen;Sun, Weilin;Yuan, Hongli
    • Journal of Microbiology and Biotechnology
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    • 제26권7호
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    • pp.1320-1332
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    • 2016
  • Light hydrocarbons accumulated in subsurface soil by long-term microseepage could favor the anomalous growth of indigenous hydrocarbon-oxidizing microorganisms, which could be crucial indicators of underlying petroleum reservoirs. Here, Illumina MiSeq sequencing of the 16S rRNA gene was conducted to determine the bacterial community structures in soil samples collected from three typical oil and gas fields at different locations in China. Incubation with n-butane at the laboratory scale was performed to confirm the presence of "universal microbes" in light-hydrocarbon microseepage ecosystems. The results indicated significantly higher bacterial diversity in next-to-well samples compared with background samples at two of the three sites, which were notably different to oil-contaminated environments. Variation partitioning analysis showed that the bacterial community structures above the oil and gas fields at the scale of the present study were shaped mainly by environmental parameters, and geographic location was able to explain only 7.05% of the variation independently. The linear discriminant analysis effect size method revealed that the oil and gas fields significantly favored the growth of Mycobacterium, Flavobacterium, and Pseudomonas, as well as other related bacteria. The relative abundance of Mycobacterium and Pseudomonas increased notably after n-butane cultivation, which highlighted their potential as biomarkers of underlying oil deposits. This work contributes to a broader perspective on the bacterial community structures shaped by long-term light-hydrocarbon microseepage and proposes relatively universal indicators, providing an additional resource for the improvement of microbial prospecting of oil and gas.

천궁 초작과 연작 재배지의 토양특성이 토양 곰팡이 군집에 미치는 영향 (Effect of Soil Properties on Soil Fungal Community in First and Continuous Cultivation Fields of Cnidium officinale Makino)

  • 김기윤;한경민;김현준;김충우;전권석;정충렬
    • 한국약용작물학회지
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    • 제28권3호
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    • pp.209-220
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    • 2020
  • Background: This study investigated the effects of soil properties on the soil fungal community in first and continuous cultivation areas of Cnidium officinale Makino. Methods and Results: The soil fungal community was analyzed for relative abundance and principal coordinate analysis (PCoA) was conducted using Illumina MiSeq sequencing. The correlation between the soil chemical properties and the soil fungal community was assessed with distance-based linear models (DISTLM). The soil fungal community showed distinct clusters consisting in the continuous cultivation area of C. officinale Makino. PCoA and DISTLM indicated that soil pH, calcium, and available P2O5 significantly affected the soil fungal community in the cultivation area of C. officinale Makino. In addition, considering 5 different pathogenic fungi the relative abundance of Fusarium in the continuous cultivation area was significantly higher compared to that in the first cultivation area of C. officinale Makino. Conclusions: This study is important because it has determinined the effects of soil properties on the soil fungal community in both first and continuous cultivation areas of C. officinale Makino. Moreover, these results will be helpful to investigate the cause of continuous cropping obstacle in C. officinale Makino by examining the changes of soil fungal community.

Metabolite Profiling and Microbial Community of Traditional Meju Show Primary and Secondary Metabolite Differences Correlated with Antioxidant Activities

  • Song, Da Hye;Chun, Byung Hee;Lee, Sunmin;Reddy, Chagam Koteswara;Jeon, Che Ok;Lee, Choong Hwan
    • Journal of Microbiology and Biotechnology
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    • 제30권11호
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    • pp.1697-1705
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    • 2020
  • Meju, a type of fermented soybean paste, is used as a starter in the preparation of various Korean traditional soybean-based foods. In this study, we performed Illumina-MiSeq paired-end sequencing for microbial communities and mass spectrometry analysis for metabolite profiling to investigate the differences between 11 traditional meju products from different regions across Korea. Even though the bacterial and fungal communities showed remarkable variety, major genera including Bacillus, Enterococcus, Variovorax, Pediococcus, Weissella, and Aspergillus were detected in every sample of meju. The metabolite profile patterns of the 11 samples were clustered into two main groups: group I (M1-5) and group II (M6-11). The metabolite analysis indicated a relatively higher amino acid content in group I, while group II exhibited higher isoflavone, soyasaponin, and lysophospholipid contents. The bioactivity analysis proved that the ABTS (2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid)) radical-scavenging activity was higher in group II and the FRAP (ferric reducing antioxidant power) activity was higher in group I. The correlation analysis revealed that the ABTS activity was isoflavonoid, lipid, and soyasaponin related, whereas the FRAP activity was amino acid and flavonoid related. These results suggest that the antioxidant activities of meju are critically influenced by the microbiome and metabolite dynamics.

Sediment Bacterial Community Structure under the Influence of Different Domestic Sewage Types

  • Zhang, Lei;Xu, Mengli;Li, Xingchen;Lu, Wenxuan;Li, Jing
    • Journal of Microbiology and Biotechnology
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    • 제30권9호
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    • pp.1355-1366
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    • 2020
  • Sediment bacterial communities are critical to the biogeochemical cycle in river ecosystems, but our understanding of the relationship between sediment bacterial communities and their specific input streams in rivers remains insufficient. In this study, we analyzed the sediment bacterial community structure in a local river receiving discharge of urban domestic sewage by applying Illumina MiSeq high-throughput sequencing. The results showed that the bacterial communities of sediments samples of different pollution types had similar dominant phyla, mainly Proteobacteria, Actinobacteria, Chloroflexi and Firmicutes, but their relative abundances were different. Moreover, there were great differences at the genus level. For example, the genus Bacillus showed statistically significant differences in the hotel site. The clustering of bacterial communities at various sites and the dominant families (i.e., Nocardioidaceae, and Sphingomonadaceae) observed in the residential quarter differed from other sites. This result suggested that environmentally induced species sorting greatly influenced the sediment bacterial community composition. The bacterial co-occurrence patterns showed that the river bacteria had a nonrandom modular structure. Microbial taxonomy from the same module had strong ecological links (such as the nitrogenium cycle and degradation of organic pollutants). Additionally, PICRUSt metabolic inference analysis showed the most important function of river bacterial communities under the influence of different types of domestic sewage was metabolism (e.g., genes related to xenobiotic degradation predominated in residential quarter samples). In general, our results emphasize that the adaptive changes and interactions in the bacterial community structure of river sediment represent responses to different exogenous pollution sources.

Characterization of microbiota diversity of engorged ticks collected from dogs in China

  • Wang, Seongjin;Hua, Xiuguo;Cui, Li
    • Journal of Veterinary Science
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    • 제22권3호
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    • pp.37.1-37.14
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    • 2021
  • Background: Ticks are one of the most common external parasites in dogs, and are associated with the transmission of a number of major zoonoses, which result in serious harm to human health and even death. Also, the increasing number of pet dogs and pet owners in China has caused concern regarding human tick-borne illnesses. Accordingly, studies are needed to gain a complete understanding of the bacterial composition and diversity of the ticks that parasitize dogs. Objectives: To date, there have been relatively few reports on the analysis of the bacterial community structure and diversity in ticks that parasitize dogs. The objective of this study was to investigate the microbial composition and diversity of parasitic ticks of dogs, and assessed the effect of tick sex and geographical region on the bacterial composition in two tick genera collected from dogs in China. Methods: A total of 178 whole ticks were subjected to a 16S ribosomal RNA (rRNA) next generation sequencing analysis. The Illumina MiSeq platform targeting the V3-V4 region of the 16S rRNA gene was used to characterize the bacterial communities of the collected ticks. Sequence analysis and taxonomic assignment were performed using QIIME 2 and the GreenGene database, respectively. After clustering the sequences into taxonomic units, the sequences were quality-filtered and rarefied. Results: After pooling 24 tick samples, we identified a total of 2,081 operational taxonomic units, which were assigned to 23 phyla and 328 genera, revealing a diverse bacterial community profile. The high, moderate and low prevalent taxa include 46, 101, and 182 genera, respectively. Among them, dominant taxa include environmental bacterial genera, such as Psychrobacter and Burkholderia. Additionally, some known tick-associated endosymbionts were also detected, including Coxiella, Rickettsia, and Ricketssiella. Also, the potentially pathogenic genera Staphylococcus and Pseudomonas were detected in the tick pools. Moreover, our preliminary study found that the differences in microbial communities are more dependent on the sampling location than tick sex in the tick specimens collected from dogs. Conclusions: The findings of this study support the need for future research on the microbial population present in ticks collected from dogs in China.

Intestinal Microbial Dysbiosis in Beagles Naturally Infected with Canine Parvovirus

  • Park, Jun Seok;Guevarra, Robin B.;Kim, Bo-Ra;Lee, Jun Hyung;Lee, Sun Hee;Cho, Jae Hyoung;Kim, Hyeri;Cho, Jin Ho;Song, Minho;Lee, Ju-Hoon;Isaacson, Richard E.;Song, Kun Ho;Kim, Hyeun Bum
    • Journal of Microbiology and Biotechnology
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    • 제29권9호
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    • pp.1391-1400
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    • 2019
  • Canine parvoviral enteritis (PVE) is an important intestinal disease of the puppies; however, the potential impact of the canine parvovirus (CPV) on the gut microbiota has not been investigated. Therefore, the aim of this study was to evaluate the gut microbial shifts in puppies naturally infected with CPV. Fecal samples were collected from healthy dogs and those diagnosed with PVE at 4, 6, 8, and 12 weeks of age. The distal gut microbiota of dogs was characterized using Illumina MiSeq sequencing of the bacterial 16S rRNA genes. The sequence data were analyzed using QIIME with an Operational Taxonomic Unit definition at a similarity cutoff of 97%. Our results showed that the CPV was associated with significant microbial dysbiosis of the intestinal microbiota. Alpha diversity and species richness and evenness in dogs with PVE decreased compared to those of healthy dogs. At the phylum level, the proportion of Proteobacteria was significantly enriched in dogs with PVE while Bacteroidetes was significantly more abundant in healthy dogs (p < 0.05). In dogs with PVE, Enterobacteriaceae was the most abundant bacterial family accounting for 36.44% of the total bacterial population compared to only 0.21% in healthy puppies. The two most abundant genera in healthy dogs were Prevotella and Lactobacillus and their abundance was significantly higher compared to that of dogs with PVE (p < 0.05). These observations suggest that disturbances of gut microbial communities were associated with PVE in young dogs. Evaluation of the roles of these bacterial groups in the pathophysiology of PVE warrants further studies.

Responses of Soil Bacterial and Fungal Communities to Organic and Conventional Farming Systems in East China

  • Zhang, Hanlin;Zheng, Xianqing;Bai, Naling;Li, Shuangxi;Zhang, Juanqin;Lv, Weiguang
    • Journal of Microbiology and Biotechnology
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    • 제29권3호
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    • pp.441-453
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    • 2019
  • Organic farming is considered an effective form of sustainable agricultural management. However, understanding of soil microbial diversity and composition under long-term organic and conventional farming is still limited and controversial. In this study, the Illumina MiSeq platform was applied to investigate the responses of soil bacterial and fungal diversity and compositions to organic farming (OF) and improved conventional farming (CF, applied straw retention) in the rice-wheat rotation system. The results highlighted that the alpha diversity of microbial communities did not differ significantly, except for higher bacterial diversity under OF. However, there were significant differences in the compositions of the soil bacterial and fungal communities between organic and conventional farming. Under our experimental conditions, through the ecological functional analysis of significant different or unique bacterial and fungal taxonomic members at the phyla and genus level, OF enhanced nitrogen, sulfur, phosphorus and carbon dynamic cycling in soil with the presence of Nodosilinea, Nitrospira, LCP-6, HB118, Lyngbya, GOUTA19, Mesorhizobium, Sandaracinobacter, Syntrophobacter and Sphingosinicella, and has the potential to strengthen soil metabolic ability with Novosphingobium. On the other hand, CF increased the intensity of nitrogen cycling with Ardenscatena, KD1-23, Iamia, Nitrosovibrio and Devosia, but enriched several pathogen fungal members, including Coniochaeta, Corallomycetella, Cyclaneusma, Cystostereum, Fistulina, Curvularia and Dissoconium.

Effect of Bacillus mesonae H20-5 Treatment on Rhizospheric Bacterial Community of Tomato Plants under Salinity Stress

  • Lee, Shin Ae;Kim, Hyeon Su;Sang, Mee Kyung;Song, Jaekyeong;Weon, Hang-Yeon
    • The Plant Pathology Journal
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    • 제37권6호
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    • pp.662-672
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    • 2021
  • Plant growth-promoting bacteria improve plant growth under abiotic stress conditions. However, their effects on microbial succession in the rhizosphere are poorly understood. In this study, the inoculants of Bacillus mesonae strain H20-5 were administered to tomato plants grown in soils with different salinity levels (EC of 2, 4, and 6 dS/m). The bacterial communities in the bulk and rhizosphere soils were examined 14 days after H20-5 treatment using Illumina MiSeq sequencing of the bacterial 16S rRNA gene. Although the abundance of H20-5 rapidly decreased in the bulk and rhizosphere soils, a shift in the bacterial community was observed following H20-5 treatment. The variation in bacterial communities due to H20-5 treatment was higher in the rhizosphere than in the bulk soils. Additionally, the bacterial species richness and diversity were greater in the H20-5 treated rhizosphere than in the control. The composition and structure of the bacterial communities varied with soil salinity levels, and those in the H20-5 treated rhizosphere soil were clustered. The members of Actinobacteria genera, including Kineosporia, Virgisporangium, Actinoplanes, Gaiella, Blastococcus, and Solirubrobacter, were enriched in the H20-5 treated rhizosphere soils. The microbial co-occurrence network of the bacterial community in the H20-5 treated rhizosphere soils had more modules and keystone taxa compared to the control. These findings revealed that the strain H20-5 induced systemic tolerance in tomato plants and influenced the diversity, composition, structure, and network of bacterial communities. The bacterial community in the H20-5 treated rhizosphere soils also appeared to be relatively stable to soil salinity changes.