• Title/Summary/Keyword: 16S-rRNA

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Characterization of Cholesterol Lowering Lactic Acid Bacteria Isolated from Palm Wine and Maize Beer and Assessment of Their Use in the Production of Probiotic Papaya Juice

  • Bertrand Tatsinkou Fossi;Dickson Ebwelle Ekabe;Liliane Laure Toukam Tatsinkou;Rene Bilingwe Ayiseh;Frederic Tavea;Pierre Michel Jazet
    • Microbiology and Biotechnology Letters
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    • v.51 no.2
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    • pp.191-202
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    • 2023
  • Elevated serum cholesterol is a main risk factor for heart disorders. Most probiotic products administered to lower cholesterol are dairy products which are not suitable for lactose-intolerant individuals. In this study, we assessed the cholesterol-lowering efficacy of LAB isolated from traditionally fermented drinks in diet-induced rats and determine their efficacy in the production of non-dairy, probiotic formulations using papaya juice. LAB were isolated from palm wine and corn beer on MRS agar using a pour-plate technique. Identification was carried out using 16S rRNA gene sequencing. A hypercholesterolemia model in which diet-induced Wistar albino rats were assigned into four groups was established. Oral gavage was carried out for 30 days. On the 31st day, the rats were dissected and the serum lipid profile was analyzed using biochemical kits. A 106 cfu/ml of a 24-h-old culture of selected lactobacilli was used to inoculate papaya juice and incubated at 37℃. Microbial and chemical changes were assessed during papaya fermentation and after four weeks of cold storage. Two selected isolates (Pw1 and Cb4) had in vitro cholesterol reduction of > 80%. These two isolates lowered lipid profile (triglyceride, total cholesterol, LDL-c) significantly, and increased HDL-c levels (p < 0.5) in the rat sera. Phylogenetic analysis showed that Pw1 was 98.86% similar to Limosilactobacillus fermentum, while Cb4 was 99.54% similar to Enteroccocus faecium. Both strains fermented papaya juice with cell viability reaching 8.92 × 108 cfu/ml and 25.3 × 108 cfu/ml respectively, and were still viable after 4 weeks of cold storage.

Biological Control of Fusarium oxysporum, the Causal Agent of Fusarium Basal Rot in Onion by Bacillus spp.

  • Jong-Hwan Shin;Ha-Kyoung Lee;Seong-Chan Lee;You-Kyoung Han
    • The Plant Pathology Journal
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    • v.39 no.6
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    • pp.600-613
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    • 2023
  • Fusarium oxysporum is the main pathogen causing Fusarium basal rot in onion (Allium cepa L.), which incurs significant yield losses before and after harvest. Among management strategies, biological control is an environmentally safe and sustainable alternative to chemical control. In this study, we isolated and screened bacteria for antifungal activity against the basal rot pathogen F. oxysporum. Isolates 23-045, 23-046, 23-052, 23-055, and 23-056 significantly inhibited F. oxysporum mycelial growth and conidial germination. Isolates 23-045, 23-046, 23-052, and 23-056 suppressed the development of Fusarium basal rot in both onion seedlings and bulbs in pot and spray inoculation assays. Isolate 23-055 was effective in onion seedlings but exhibited weak inhibitory effect on onion bulbs. Based on analyses of the 16S rRNA and rpoB gene sequences together with morphological analysis, isolates 23-045, 23-046, 23-052, and 23-055 were identified as Bacillus thuringiensis, and isolate 23-056 as Bacillus toyonensis. All five bacterial isolates exhibited cellulolytic, proteolytic, and phosphate-solubilizing activity, which may contribute to their antagonistic activity against onion basal rot disease. Taken together B. thuringiensis 23-045, 23-046, 23-052, and 23-055 and B. toyonensis 23-056 have potential for the biological control of Fusarium basal rot in onion.

Abundance and Diversity of Microbial Communities in the Coastal Aquifers in Songji Lagoon, South Korea (송지호 해안 대수층 미생물 군집의 풍부도 및 다양성)

  • Jung-Yun Lee;Dong-Hun Kim;Woo-Hyun Jeon;Hee Sun Moon
    • Journal of Soil and Groundwater Environment
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    • v.28 no.5
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    • pp.12-24
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    • 2023
  • The Songji lagoon is brackish environment with a mixture of saline and fresh water, and the interaction of groundwater-lagoon water creates a physicochemical gradient. Although some studies have been conducted on the hydrological and geochemical characteristics of the Songji lagoon, microbial ecological studies have not yet been conducted. In this study, we investigated the effect of groundwater and surface water interaction on water quality as well as microbial community changes in the Songji Lagoon using 16S rRNA gene sequencing. Hydrochemical analyses show that samples were classified as 5 hydrochemical facies (HF) and hydrochemical facies evolution (HFE) revealed the intrusion phase was more dominant (57.9%) than the freshening phase (42.1%). Higher microbial diversity was found in freshwater in comparison to saline water samples. The microbial community at the phylum level shows the most dominance of Proteobacteria with an average of 37.3%, followed by Bacteroidota, Actinobacteria, and Patescibacteria. Heat map analyses of the top 18 genera showed that samples were clustered into 5 groups based on type, and Pseudoalteromonas could be used potential indicator for seawater intrusion.

Isolation of the Bacteria Strains with Highly Active β-glucosidase from Traditional Wine and Use them for the Synthesis of Non G lucoside Product in Plant (식물유래 비배당체 활성 성분 증대를 위한 고활성 β-glucosidase 생산 균주 선별 및 특성 규명)

  • Tae-Su Kim;Sua Im;Do Yun Jeong;Byung Hee Chun;Yun Ji Kim;Myoung Lae Cho;Bo-Ram Kim;Su Hui Seong;Jin-Woo Jeong
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2022.09a
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    • pp.125-125
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    • 2022
  • 식물 내에는 배당체 형태의 생리활성 물질들이 다량 존재하고 있으며, 비배당체로 전환하였을 경우 생리활성이나 투과성이 증대된다는 연구들이 보고되어 있다. 따라서 본 연구에서는 고활성 β-glucosidase 활성을 갖는 균주를 선별함으로써 식물 내의 비배당체 활성 성분을 증대시키는 데 활용하고자 한다. 균주는 전남지역 제조된 막걸리부터 분리하였다. 분리된 균주는 성장 속도 등 산업적으로 활용될 수 있는 가치를 평가하여 11종 22점의 균주를 분리하고, 16S rRNA분석을 통해 균주를 동정하였다. 동정된 균주를 이용하여 β-glucosidase의 활성을 평가하여 고활성 균주를 1종을 선별하였다. 선별된 균주는 다른 균주에 비하여 상대적 활성이 57% ~ 99%정도 활성이 높았으며, 공시 균주와 비교하였을 때도 상대 활성이 높은 았다. 선별된 균주의 특성을 규명하기 위하여 glycerol, L-arbinose를 비롯한 49가지의 탄수화물의 대사 활성을 확인해본 결과 25가지의 탄수화물 대사 활성을 가지고 있는 것을 알수 있었다. 또한 Lipase, β-glucosidase 19가지의 다양한 효소활성을 확인해본 결과 8종의 효소활성이 있음을 확인할 수 있었다.

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Evaluation of Antibiotics Resistance for Human-harmful Bacteria Isolated from Eco-friendly and Practical Cultivation Farms of Hot Pepper and Tomato (고추 및 토마토 친환경 및 관행재배지에서 분리한 인체 유해세균의 항생제 저항성 평가)

  • Lee, Sung-Hee;Do, Jiwon;Kim, Seong Kyeom;Oh, Kwang Kyo;Park, Jae-Ho
    • Korean Journal of Organic Agriculture
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    • v.31 no.4
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    • pp.381-394
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    • 2023
  • This study was conducted to monitor the antibiotics resistance of human-harmful bacteria isolated in the agricultural environment for hot peppers (Capsicum annuum) and tomato (Lycopersicon esculentum). As a result, we isolated 120 bacterial species (34 on fruits, 48 in soil, 21 in water, and 17 in manure), identified them with the 16S rRNA sequence, analyzed minimum inhibitory concentration (MIC) for 26 antibiotics using Sensititre ARIS Hi-Q system and then evaluated whether each bacterial genus acquired resistance for the tested antibiotics or not, according to the CLSI criteria. From difference in MIC between eco-friendly (EFM) and practical (PFM) cultivation farms, Klebsiella spp. isolated from EFM was resistant to ampicillin (AMP) and nalidixic acid (NAL), and that isolated from PFM was resistant to streptomycin (STR) and tetracycline (TET). Enterobacter spp. isolated from EFM was resistant to AMP and azithromycin (AZI), and that isolated from PFM was resistant to AMP, AZI, and STR. Meanwhile, Pseudomonas spp. isolated from EFM and PFM were all resistant to AMP, AZI, cefotaxime (FOT), cefoxitin (FOX), ceftriaxone (AXO), CHL, NAL, and STR. Staphylococcus spp. isolated from EFM and PFM were resistant to gentamycin (GEN), STR, and kanamycin (KAN), and in particular, that from EFM showed resistance for erythromycin (ERY). In conclusion, our study suggested that EFM lead STR antibiotics resistance for human-harmful bacteria to decrease, because only the bacteria isolated from hot pepper and tomato crop with PFM have showed resistance against STR antibiotics, regardless of bacterial genus.

Subtype-Based Microbial Analysis in Non-small Cell Lung Cancer

  • Hye Jin Jang;Eunkyung Lee;Young-Jae Cho;Sang Hoon Lee
    • Tuberculosis and Respiratory Diseases
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    • v.86 no.4
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    • pp.294-303
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    • 2023
  • Background: The human lung serves as a niche for a unique and dynamic bacterial community related to the development and aggravation of multiple respiratory diseases. Therefore, identifying the microbiome status is crucial to maintaining the microecological balance and maximizing the therapeutic effect on lung diseases. Therefore, we investigated the histological type-based differences in the lung microbiomes of patients with lung cancer. Methods: We performed 16S rRNA sequencing to evaluate the respiratory tract microbiome present in bronchoalveolar lavage fluid. Patients with non-small cell lung cancer were stratified based on two main subtypes of lung cancer: adenocarcinoma and squamous cell carcinoma (SqCC). Results: Among the 84 patients analyzed, 64 (76.2%) had adenocarcinoma, and 20 (23.8%) had SqCC. The α- and β-diversities showed significant differences between the two groups (p=0.004 for Chao1, p=0.001 for Simpson index, and p=0.011 for PERMANOVA). Actinomyces graevenitzii was dominant in the SqCC group (linear discriminant analysis [LDA] score, 2.46); the populations of Haemophilus parainfluenza (LDA score, 4.08), Neisseria subflava (LDA score, 4.07), Porphyromonas endodontalis (LDA score, 3.88), and Fusobacterium nucleatum (LDA score, 3.72) were significantly higher in the adenocarcinoma group. Conclusion: Microbiome diversity is crucial for maintaining homeostasis in the lung environment, and dysbiosis may be related to the development and prognosis of lung cancer. The mortality rate was high, and the microbiome was not diverse in SqCC. Further large-scale studies are required to investigate the role of the microbiome in the development of different lung cancer types.

Comparative analysis of the pig gut microbiome associated with the pig growth performance

  • Jun Hyung Lee;San Kim;Eun Sol Kim;Gi Beom Keum;Hyunok Doo;Jinok Kwak;Sriniwas Pandey;Jae Hyoung Cho;Sumin Ryu;Minho Song;Jin Ho Cho;Sheena Kim;Hyeun Bum Kim
    • Journal of Animal Science and Technology
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    • v.65 no.4
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    • pp.856-864
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    • 2023
  • There are a variety of microorganisms in the animal intestine, and it has been known that they play important roles in the host such as suppression of potentially pathogenic microorganisms, modulation of the gut immunity. In addition, the gut microbiota and the livestock growth performance have long been known to be related. Therefore, we evaluated the interrelation between the growth performance and the gut microbiome of the pigs from 3 different farms, with pigs of varied ages ready to be supplied to the market. When pigs reached average market weight of 118 kg, the average age of pigs in three different farms were < 180 days, about 190 days, and > 200 days, respectively. Fecal samples were collected from pigs of age of 70 days, 100 days, 130 days, and 160 days. The output data of the 16S rRNA gene sequencing by the Illumina Miseq platform was filtered and analyzed using Quantitative Insights into Microbial Ecology (QIIME)2, and the statistical analysis was performed using Statistical Analysis of Metagenomic Profiles (STAMP). The results of this study showed that the gut microbial communities shifted as pigs aged along with significant difference in the relative abundance of different phyla and genera in different age groups of pigs from each farm. Even though, there was no statistical differences among groups in terms of Chao1, the number of observed operational taxonomic units (OTUs), and the Shannon index, our results showed higher abundances of Bifidobacterium, Clostridium and Lactobacillus in the feces of pigs with rapid growth rate. These results will help us to elucidate important gut microbiota that can affect the growth performance of pigs.

Association between LEPR Genotype and Gut Microbiome in Healthy Non-Obese Korean Adults

  • Yoon Jung Cha;In Ae Chang;Eun-Heui Jin;Ji Hye Song;Jang Hee Hong;Jin-Gyu Jung;Jung Sunwoo
    • Biomolecules & Therapeutics
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    • v.32 no.1
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    • pp.146-153
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    • 2024
  • The LEPR (leptin receptor) genotype is associated with obesity. Gut microbiome composition differs between obese and non-obese adults. However, the impact of LEPR genotype on gut microbiome composition in humans has not yet been studied. In this study, the association between LEPR single nucleotide polymorphism (rs1173100, rs1137101, and rs790419) and the gut microbiome composition in 65 non-obese Korean adults was investigated. Leptin, triglyceride, low-density lipoprotein cholesterol, and high-density lipoprotein cholesterol levels were also measured in all participants. Mean ± SD (standard deviation) of age, body mass index, and leptin hormone levels of participants was 35.2 ± 8.1 years, 21.4 ± 1.8 kg/m2, and 7989.1 ± 6687.4 pg/mL, respectively. Gut microbiome analysis was performed at the phylum level by 16S rRNA sequencing. Among the 11 phyla detected, only one showed significantly different relative abundances between LEPR genotypes. The relative abundance of Candidatus Saccharibacteria was higher in the G/A genotype group than in the G/G genotype group for the rs1137101 single nucleotide polymorphism (p=0.0322). Participant characteristics, including body mass index, leptin levels, and other lipid levels, were similar between the rs1137101 G/G and G/A genotypes. In addition, the relative abundances of Fusobacteria and Tenericutes showed significant positive relationship with plasma leptin concentrations (p=0.0036 and p=0.0000, respectively). In conclusion, LEPR genotype and gut microbiome may be associated even in normal-weight Korean adults. However, further studies with a greater number of obese adults are needed to confirm whether LEPR genotype is related to gut microbiome composition.

Nicotinic acid changes rumen fermentation and apparent nutrient digestibility by regulating rumen microbiota in Xiangzhong black cattle

  • Zhuqing Yang;Linbin Bao;Wanming Song;Xianghui Zhao;Huan Liang;Mingjin Yu;Mingren Qu
    • Animal Bioscience
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    • v.37 no.2
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    • pp.240-252
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    • 2024
  • Objective: The aim of this study was to investigate the impact of dietary nicotinic acid (NA) on apparent nutrient digestibility, rumen fermentation, and rumen microbiota in uncastrated Xiangzhong black cattle. Methods: Twenty-one uncastrated Xiangzhong black cattle (385.08±15.20 kg) aged 1.5 years were randomly assigned to the control group (CL, 0 mg/kg NA in concentrate diet), NA1 group (800 mg/kg NA in concentrate diet) and NA2 group (1,200 mg/kg NA in concentrate diet). All animals were fed a 60% concentrate diet and 40% dried rice straw for a 120-day feeding experiment. Results: Supplemental NA not only enhanced the apparent nutrient digestibility of acid detergent fiber (p<0.01), but also elevated the rumen acetate and total volatile fatty acid concentrations (p<0.05). 16S rRNA gene sequencing analysis of rumen microbiota revealed that dietary NA changed the diversity of rumen microbiota (p<0.05) and the abundance of bacterial taxa in the rumen. The relative abundances of eight Erysipelotrichales taxa, five Ruminococcaceae taxa, and five Sphaerochaetales taxa were decreased by dietary NA (p<0.05). However, the relative abundances of two taxa belonging to Roseburia faecis were increased by supplemental 800 mg/kg NA, and the abundances of seven Prevotella taxa, three Paraprevotellaceae taxa, three Bifidobacteriaceae taxa, and two operational taxonomic units annotated to Fibrobacter succinogenes were increased by 1,200 mg/kg NA in diets. Furthermore, the correlation analysis found significant correlations between the concentrations of volatile fatty acids in the rumen and the abundances of bacterial taxa, especially Prevotella. Conclusion: The results from this study suggest that dietary NA plays an important role in regulating apparent digestibility of acid detergent fiber, acetate, total volatile fatty acid concentrations, and the composition of rumen microbiota.

Comparative Genome analysis of the Genus Curvibacter and the Description of Curvibacter microcysteis sp. nov. and Curvibacter cyanobacteriorum sp. nov., Isolated from Fresh Water during the Cyanobacterial Bloom Period

  • Ve Van Le;So-Ra Ko;Mingyeong Kang;Seonah Jeong;Hee-Mock Oh;Chi-Yong Ahn
    • Journal of Microbiology and Biotechnology
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    • v.33 no.11
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    • pp.1428-1436
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    • 2023
  • The three Gram-negative, catalase- and oxidase-positive bacterial strains RS43T, HBC28, and HBC61T, were isolated from fresh water and subjected to a polyphasic study. Comparison of 16S rRNA gene sequence initially indicated that strains RS43T, HBC28, and HBC61T were closely related to species of genus Curvibacter and shared the highest sequence similarity of 98.14%, 98.21%, and 98.76%, respectively, with Curvibacter gracilis 7-1T. Phylogenetic analysis based on genome sequences placed all strains within the genus Curvibacter. The average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) values between the three strains and related type strains supported their recognition as two novel genospecies in the genus Curvibacter. Comparative genomic analysis revealed that the genus possessed an open pangenome. Based on KEGG BlastKOALA analyses, Curvibacter species have the potential to metabolize benzoate, phenylacetate, catechol, and salicylate, indicating their potential use in the elimination of these compounds from the water systems. The results of polyphasic characterization indicated that strain RS43T and HBC61T represent two novel species, for which the name Curvibacter microcysteis sp. nov. (type strain RS43T =KCTC 92793T=LMG 32714T) and Curvibacter cyanobacteriorum sp. nov. (type strain HBC61T =KCTC 92794T=LMG 32713T) are proposed.