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1.
Environ Monit Assess ; 196(4): 341, 2024 Mar 04.
Article in English | MEDLINE | ID: mdl-38436747

ABSTRACT

Bacterial communities in epilithic biofilm plays an important role in biogeochemistry processes in freshwater ecosystems. Nevertheless, our understanding of the geographical and seasonal variations of the composition of bacterial communities in the biofilm of gravels on river bed is still limited. Various anthropogenic activities also influence the biofilm bacteria in gravel rivers. By taking the Shiting River in the upper Yangtze River basin in Sichuan Province as an example, we studied the geographical and seasonal variations of epilithic bacteria and the impacts of weirs and other human activities (e.g., sewage pollution). The river has experienced severe degradation since the Ms 8.0 Wenchuan Earthquake, and weirs were constructed to prevent bed erosion. We collected epilithic biofilms samples at 17 sites along ~ 30 km river reach of the Shiting River in the autumn of 2021 and the summer of 2022, respectively. We applied 16S rRNA gene high-throughput sequencing technology and Functional Annotation of Prokaryotic Taxa (FAPROTAX) to analyze the seasonal and biogeographic patterns and potential functions of the biofilm bacterial communities. The results showed that epilithic bacteria from the two surveys exhibited variation in community composition, bacterial diversity and potential functions. The bacteria samples collected in the autumn have much higher alpha diversity and richness than those collected in the summer. Bacterial richness and diversity were lower downstream of the weirs than upstream. Low diversity was observed at a sampling site influenced by sewage inflow, which contains high level of nitrogen-related chemicals.


Subject(s)
Anthropogenic Effects , Ecosystem , Humans , Seasons , RNA, Ribosomal, 16S , Sewage , Environmental Monitoring , Bacteria/genetics , Biofilms , China
2.
BMC Microbiol ; 23(1): 281, 2023 10 03.
Article in English | MEDLINE | ID: mdl-37784018

ABSTRACT

BACKGROUND: Angelica polysaccharides (AP) have numerous benefits in relieving type 2 diabetes (T2D). However, the underlying mechanisms have yet to be fully understood. Recent many reports have suggested that altering gut microbiota can have adverse effects on the host metabolism and contribute to the development of T2D. Here, we successfully established the T2D model using the male KKAy mice with high-fat and high-sugar feed. Meanwhile, the male C57BL/6 mice were fed with a normal feed. T2D KKAy mice were fed either with or without AP supplementation. In each group, we measured the mice's fasting blood glucose, weight, and fasting serum insulin levels. We collected the cecum content of mice, the gut microbiota was analyzed by targeted full-length 16S rRNA metagenomic sequencing and metabolites were analyzed by untargeted-metabolomics. RESULTS: We found AP effectively alleviated glycemic disorders of T2D KKAy mice, with the changes in gut microbiota composition and function. Many bacteria species and metabolites were markedly changed in T2D KKAy mice and reversed by AP. Additionally, 16 altered metabolic pathways affected by AP were figured out by combining metagenomic pathway enrichment analysis and metabolic pathway enrichment analysis. The key metabolites in 16 metabolic pathways were significantly associated with the gut microbial alteration. Together, our findings showed that AP supplementation could attenuate the diabetic phenotype. Significant gut microbiota and gut metabolite changes were observed in the T2D KKAy mice and AP intervention. CONCLUSIONS: Administration of AP has been shown to improve the composition of intestinal microbiota in T2D KKAy mice, thus providing further evidence for the potential therapeutic application of AP in the treatment of T2D.


Subject(s)
Diabetes Mellitus, Type 2 , Gastrointestinal Microbiome , Mice , Male , Animals , Gastrointestinal Microbiome/genetics , Diabetes Mellitus, Type 2/drug therapy , Diabetes Mellitus, Type 2/microbiology , Blood Glucose/metabolism , RNA, Ribosomal, 16S/genetics , Mice, Inbred C57BL , Polysaccharides/pharmacology
3.
Biofouling ; 34(10): 1161-1174, 2018 11.
Article in English | MEDLINE | ID: mdl-30740996

ABSTRACT

Deterioration of concrete is a large societal cost. In the Oslofjord subsea tunnel (Norway), deterioration of sprayed concrete and corrosion of reinforcing steel fibres occur under biofilm formed at sites with intrusion of saline groundwater. In this study, the microbial community structure, in situ environmental gradients and chemical composition of the biofilms were examined at three tunnel sites. Ammonia- and nitrite-oxidising microorganisms, in particular Nitrosopumilus sp., and iron-oxidising bacteria within Mariprofundus sp., were omnipresent, together with a diversity of presumably heterotrophic bacteria. Alpha- and beta diversity measures showed significant differences in richness and community structure between the sites as well as over time and null-models suggested that deterministic factors were important for the community assembly. The superficial flow of water over the biofilm had a strong effect on oxygen penetration in the biofilm and was identified as one major environmental gradient that varied between the sites, likely being important for shaping the microbial communities.


Subject(s)
Biofilms/growth & development , Construction Materials/microbiology , Groundwater/microbiology , Microbiota/physiology , Seawater/microbiology , Steel , Archaea/isolation & purification , Corrosion , Norway , Proteobacteria/isolation & purification , RNA, Ribosomal, 16S , Steel/chemistry
4.
BMC Microbiol ; 17(1): 78, 2017 03 31.
Article in English | MEDLINE | ID: mdl-28359329

ABSTRACT

BACKGROUND: Sample collection for gut microbiota analysis from in-patients can be challenging. Collection method and storage conditions are potential sources of variability. In this study, we compared the bacterial microbiota from stool stored under different conditions, as well as stool and swab samples, to assess differences due to sample storage conditions and collection method. METHODS: Using bacterial 16S rRNA gene sequence analysis, we compared the microbiota profiles of stool samples stored and collected under various conditions. Stool samples (2 liquid, 1 formed) from three different patients at two hospitals were each evaluated under the following conditions: immediately frozen at -80°C, stored at 4°C for 12-48 hours before freezing at -80°C and stored at -20°C with 1-2 thaw cycles before storage at -80°C. Additionally, 8 stool and 30 rectal swab samples were collected from 8 in-patients at one hospital. Microbiota differences were assessed using the Yue and Clayton dissimilarity index (θYC distance) and analysis of molecular variance (AMOVA). RESULTS: Regardless of the storage conditions, the bacterial communities of aliquots from the same stool samples were very similar based on θYC distances (median intra-sample θYC distance: 0.035, IQR: 0.015-0.061) compared to aliquots from different stool samples (median inter-sample θYC distance: 0.93, IQR: 0.85-0.97) (Wilcoxon test p-value: <0.0001). For the stool and rectal swab comparison, samples from different patients, regardless of sample collection method, were significantly different (AMOVA p-values: <0.001-0.029) compared to no significant difference between all stool and swab samples (AMOVA p-value: 0.976). The θYC dissimilarity index between swab and stool samples was significantly lower within individuals (median 0.17, IQR: 0.10-0.27) than between individuals (median 0.93, IQR: 0.85-0.97) (Wilcoxon test p-value: <0.0001), indicating minimal differences between stool and swab samples collected from the same individual over the sampling period. CONCLUSION: For gastrointestinal microbiota studies based on bacterial 16S rRNA gene sequence analysis, interim stool sample storage at 4 °C or -20 °C, rather than immediate storage at -80 °C, does not significantly alter results. Additionally, stool and rectal swab microbiotas from the same subject were highly similar, indicating that these sampling methods could be used interchangeably to assess the community structure of the distal GI tract.


Subject(s)
Bacteria/classification , Feces/microbiology , Gastrointestinal Microbiome , Specimen Handling/methods , Analysis of Variance , Bacteria/genetics , Bacteria/isolation & purification , Chicago , DNA, Bacterial/genetics , Female , Gastrointestinal Microbiome/genetics , Gastrointestinal Tract/microbiology , Hospitals , Humans , Male , Middle Aged , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Temperature , Time Factors
5.
Antonie Van Leeuwenhoek ; 110(6): 811-817, 2017 Jun.
Article in English | MEDLINE | ID: mdl-28303399

ABSTRACT

A Gram-positive, non-motile and coccoid strain, designated XY-FW106T, was isolated from a marine flatworm identified to be Planocera sp. The 16S rRNA gene sequence of this pink organism was consistent with membership of the genus Deinococcus, with high sequence similarity to Deinococcus aetherius ST0316T (94.7%). The optimum growth temperature range of the strain XY-FW106T was found to be 25-30 °C and optimum growth occurs at pH 7.2-7.4 without NaCl. The strain XY-FW106T was found to contain unidentified glycolipids, unidentified phosphoglycolipids, unidentified phospholipids and unidentified lipids, which differed from those of closely related species. Menaquinone MK-8 was identified as the major respiratory quinone and the predominant cellular fatty acids were found to be Summed Feature 3 (C16:1 ω7c/C16:1 ω6c), C16:0, iso-C15:0, and Summed Feature 8 (C18:1 ω7c/C18:1 ω6c). The DNA G+C content was determined to be 70.2 mol%. The biochemical and chemotaxonomic data together suggest that the strain represents a new species for which the name Deinococcus planocerae sp. nov. is proposed. The type strain is XY-FW106T (=MCCC 1K01499T=KCTC 33809T).


Subject(s)
Deinococcus/isolation & purification , Platyhelminths/microbiology , RNA, Ribosomal, 16S , Animals , Bacterial Typing Techniques , Base Composition , DNA, Bacterial , Deinococcus/genetics , Fatty Acids , Phospholipids , Phylogeny , Sequence Analysis, DNA
6.
Microb Ecol ; 72(3): 514-25, 2016 10.
Article in English | MEDLINE | ID: mdl-27412380

ABSTRACT

Didymosphenia geminata (Lyngbye) M. Schmidt is a stalked freshwater diatom that is expanding its range globally. In some rivers, D. geminata forms thick and expansive polysaccharide-dominated mats. Like other stalked diatoms, D. geminata cells attach to the substratum with a pad of adhesive extracellular polymeric substance. Research on D. geminata and other diatoms suggests that bacterial biofilm composition may contribute to successful attachment. The aim of this study was to investigate the composition and role of bacterial biofilm communities in D. geminata attachment and survival. Bacterial biofilms were collected at four sites in the main stem of a river (containing D. geminata) and in four tributaries (free of D. geminata). Samples were characterised using automated rRNA intergenic spacer analysis and high-throughput sequencing (HTS). Mat-associated bacteria were isolated and their effect on the early establishment of D. geminata cells assessed using co-culturing experiments. ARISA and HTS data showed differences in bacterial communities between samples with and without D. geminata at two of the four sites. Samples with D. geminata had a higher relative abundance of Sphingobacteria (p < 0.01) and variability in community composition was reduced. Analysis of the 76 bacteria isolated from the mat revealed 12 different strains representing 8 genera. Co-culturing of a Carnobacterium sp. with D. geminata reduced survival (p < 0.001) and attachment (p < 0.001) of D. geminata. Attachment was enhanced by Micrococcus sp. and Pseudomonas sp. (p < 0.001 and p < 0.01, respectively). These data provide evidence that bacteria play a role in the initial attachment and on-going survival of D. geminata, and may partly explain observed distribution patterns.


Subject(s)
Bacteria/genetics , Biofilms , Coculture Techniques , Diatoms/genetics , Diatoms/microbiology , Fresh Water/microbiology , Bacteria/classification , Bacteria/growth & development , Bacteria/isolation & purification , Bacterial Physiological Phenomena/genetics , Base Sequence , Carnobacterium/growth & development , Cell Adhesion , DNA, Bacterial , Diatoms/growth & development , Diatoms/physiology , High-Throughput Nucleotide Sequencing , New Zealand , Phylogeny , RNA, Ribosomal, 16S/genetics , Rivers/microbiology , Sphingobacterium , Water Microbiology
7.
Anaerobe ; 32: 77-86, 2015 Apr.
Article in English | MEDLINE | ID: mdl-25557787

ABSTRACT

One hundred and seven mesophilic isolates of Clostridium were isolated from agricultural soils cultivated with different plants in Assuit Governorate, Egypt. Eighty isolates (out of 107) showed the ability to produce ABE (Acetone, butanol and ethanol) on T6 medium ranging from 0.036 to 31.89 g/L. The highest numbers of ABE producing isolates were obtained from soil samples of potato contributing 27 isolates, followed by 18 isolates from wheat and 10 isolates from onion. On the other hand, there were three native isolates that produced ABE more than those produced by the reference isolate Clostridium acetobutylicum ATCC 824 (11.543 g/L). The three isolates were identified based on phenotypic and gene encoding 16S rRNA as Clostridium beijerinckii ASU10 (KF372577), Clostridium chauvoei ASU55 (KF372580) and Clostridium roseum ASU58 (KF372581). The highest ABE level from substandard and surplus dates was produced by C. beijerinckii ASU10 (24.07 g/L) comprising butanol 67.15% (16.16 g/L), acetone 30.73% (7.4 g/L) and ethanol 2.12% (0.51 g/L), while C. roseum ASU58 and C. chauvoei ASU55 produced ABE contributing 20.20 and 13.79 g/L, respectively. ABE production by C. acetobutylicum ATCC 824 was 15.01 g/L. This study proved that the native strains C. beijerinckii ASU10 and C. roseum ASU58 have high competitive efficacy on ABE production from economical substrate as substandard and surplus date fruits. Additionally, using this substrate without any nutritional components is considered to be a commercial substrate for desired ABE production.


Subject(s)
Acetone/metabolism , Butanols/metabolism , Clostridium/metabolism , Ethanol/metabolism , Fermentation , Biofuels , Clostridium/classification , Clostridium/genetics , Clostridium/isolation & purification , Genotype , Phenotype , Phylogeny , RNA, Ribosomal, 16S/genetics , Soil Microbiology
8.
J Basic Microbiol ; 55(8): 1013-20, 2015 Aug.
Article in English | MEDLINE | ID: mdl-25871924

ABSTRACT

16S rRNA and nifD-nifK sequences were used to study the molecular phylogeny and evolutionary genetics of Frankia strains isolated from Hippöphae salicifolia D. Don growing at different altitudes (ecologically classified as riverside and hillside isolates) of the Eastern Himalayan region of North Sikkim, India. Genetic information for the small subunit rRNA (16S rRNA) revealed that the riverside Frankia isolates markedly differed from the hillside isolates suggesting that the riverside isolates are genetically compact. Further, for enhanced resolutions, the partial sequence of nifD (3' end), nifK (5' end) and nifD-K IGS region have been investigated. The sequences obtained, failed to separate riverside isolates and hillside isolates, thus suggesting a possible role of genetic transfer events either from hillside to riverside or vice versa. The evolutionary genetic analyses using evogenomic extrapolations of gene sequence data obtained from 16S rRNA and nifD-K provided differing equations with the pace of evolution being more appropriately, intermediate. Values of recombination frequency (R), nucleotide diversity per site (Pi), and DNA divergence estimates supported the existence of an intermixed zone where spatial isolations occurred in sync with the temporal estimates. J. Basic Microbiol. 2015, 54, 1-9.


Subject(s)
Evolution, Molecular , Frankia/genetics , Phylogeny , RNA, Ribosomal, 16S/genetics , DNA, Intergenic , Frankia/classification , Frankia/growth & development , Genes, Bacterial , Genes, rRNA , Hippophae/microbiology , India , Nitrogenase/genetics , Polymerase Chain Reaction , Root Nodules, Plant/microbiology , Sequence Analysis, DNA
9.
Front Microbiol ; 15: 1429035, 2024.
Article in English | MEDLINE | ID: mdl-39104582

ABSTRACT

The alarming rise in antimicrobial resistance (AMR) has created a significant public health challenge, necessitating the discovery of new therapeutic agents to combat infectious diseases and oxidative stress-related disorders. The Lentzea flaviverrucosa strain E25-2, isolated from Moroccan forest soil, represents a potential avenue for such research. This study aimed to identify the isolate E25-2, obtained from soil in a cold Moroccan ecosystem, and further investigate its antimicrobial and antioxidant activities. Phylogenetic analysis based on 16S rRNA gene sequences revealed the strain's classification within the Lentzea genus, with a sequence closely resembling that of Lentzea flaviverrucosa AS4.0578 (96.10% similarity). Antimicrobial activity in solid media showed moderate to strong activity against Staphylococcus aureus ATCC 25923, Bacillus cereus strain ATCC 14579, Escherichia coli strain ATCC 25922, Candida albicans strain ATCC 60193 and 4 phytopathogenic fungi. In addition, ethyl acetate extract of this isolate demonstrated potent antimicrobial activity against 7 clinically multi-drug resistant bacteria. Furthermore, it demonstrated antioxidant activity against 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) free radicals, as well as a significant increase in ferric reducing antioxidant power. A significant positive correlation was observed between antioxidant activities and total content of phenolic compounds (p < 0.0001), along with flavonoids (p < 0.0001). Furthermore, gas chromatography-mass spectrometry (GC-MS) analysis revealed the presence of amines, hydroxyl groups, pyridopyrazinone rings, esters and pyrrolopyrazines. The Lentzea genus could offer promising prospects in the fight against antibiotic resistance and in the prevention against oxidative stress related diseases.

10.
Stem Cell Res Ther ; 13(1): 475, 2022 09 14.
Article in English | MEDLINE | ID: mdl-36104756

ABSTRACT

BACKGROUND: Crohn's disease (CD) is a chronic non-specific inflammatory bowel disease. Current CD therapeutics cannot fundamentally change the natural course of CD. Therefore, it is of great significance to find new treatment strategies for CD. Preclinical and clinical studies have shown that mesenchymal stromal cells (MSCs) are a promising therapeutic approach. However, the mechanism by which MSCs alleviate CD and how MSCs affect gut microbes are still unclear and need further elucidation. METHODS: We used 2,4,6-trinitrobenzenesulfonic acid (TNBS) to induce experimental colitis in mice and analysed the microbiota in faecal samples from the control group, the TNBS group and the TNBS + MSC group with faecal 16S rDNA sequencing. Subsequent analyses of alpha and beta diversity were all performed based on the rarified data. PICRUStII analysis was performed on the 16S rRNA gene sequences to infer the gut microbiome functions. RESULTS: MSC Treatment improved TNBS-induced colitis by increasing survival rates and relieving symptoms. A distinct bacterial signature was found in the TNBS group that differed from the TNBS + MSC group and controls. MSCs prevented gut microbiota dysbiosis, including increasing α-diversity and the amount of Bacteroidetes Firmicutes and Tenericutes at the phylum level and decreasing the amount of Proteobacteria at the phylum level. MSCs alleviated the increased activities of sulphur and riboflavin metabolism. Meanwhile some metabolic pathways such as biosynthesis of amino acids lysine biosynthesis sphingolipid metabolism and secondary bile acid biosynthesis were decreased in the TNBS group compared with the control group and the TNBS + MSC group CONCLUSIONS: Overall, our findings preliminarily confirmed that colitis in mice is closely related to microbial and metabolic dysbiosis. MSC treatment could modulate the dysregulated metabolism pathways in mice with colitis, restoring the abnormal microbiota function to that of the normal control group. This study provides insight into specific intestinal microbiota and metabolism pathways linked with MSC treatment, suggesting a new approach to the treatment of CD.


Subject(s)
Colitis , Crohn Disease , Gastrointestinal Microbiome , Mesenchymal Stem Cells , Animals , Colitis/chemically induced , Colitis/metabolism , Colitis/therapy , Crohn Disease/therapy , Disease Models, Animal , Dysbiosis/therapy , Humans , Mesenchymal Stem Cells/metabolism , Mice , RNA, Ribosomal, 16S/genetics , Trinitrobenzenesulfonic Acid , Umbilical Cord/metabolism
11.
Front Immunol ; 13: 938326, 2022.
Article in English | MEDLINE | ID: mdl-36091072

ABSTRACT

Organisms are colonized by microorganism communities and play a pivotal role in host function by influencing physiology and development. In mammals, bacterial community may alter gonadal maturation and drive sex-specific differences in gene expression and metabolism. However, bacterial microbiota diversity in the gonads of early vertebrates has not been fully elucidated. Here, we focused on the swamp eel (Monopterus albus), which naturally undergoes sex reversal, and systematically analyzed the bacterial microbiota profiles between females and males using 16S rRNA gene sequences. Specifically, the microbial abundance and community diversity of gonads in males were higher than in females. Although Proteobacteria, Firmicutes, Bacteroidetes, and Actinobacteria were characterized as the dominating phyla in ovary and testis, the relative abundance of Firmicutes was significantly higher in males than females. Detailed analysis of the microbial community revealed that Bacilli were the dominant bacteria in ovaries and Clostridium in testes of M. albus. More importantly, we proposed that differences in the microbial composition and distribution between ovaries and testes may be linked to functional categories in M. albus, especially metabolism. These findings represent a unique resource of bacterial community in gonads to facilitate future research about the mechanism of how microbiota influence sex-specific differences and sex reversal in vertebrates.


Subject(s)
Microbiota , Smegmamorpha , Animals , Bacteria/metabolism , Female , Male , Mammals/genetics , Ovary , RNA, Ribosomal, 16S/genetics , Smegmamorpha/genetics
12.
J Microbiol ; 56(7): 478-484, 2018 Jul.
Article in English | MEDLINE | ID: mdl-29948823

ABSTRACT

A non-motile, pink-pigmented bacterial strain designated IMCC25679T, was isolated from freshwater Lake Chungju of Korea. Phylogenetic trees based on 16S rRNA gene sequences showed that the strain IMCC25679T formed a lineage within the genus Pedobacter. The strain IMCC25679T was closely related to Pedobacter daechungensis Dae 13T (96.4% sequence similarity), Pedobacter rivuli HME8457T (95.3%) and Pedobacter lentus DS-40T (94.3%). The major fatty acids of IMCC- 25679T were iso-C15:0, iso-C16:0 and summed feature 3 (comprising C16:1ω6c and/or C16:1ω7c). The major respiratory quinone was MK-7. The major polar lipids were phosphatidylethanolamine (PE), an unidentified sphingolipid (SL), an unidentified aminolipid (AL) and three unidentified polar lipids (PL). The DNA G + C content of IMCC25679T was 32.2 mol%. Based on the evidence presented in this study, the strain IMCC25679T represents a novel species within the genus Pedobacter, with the proposed name Pedobacter aquicola, sp. nov. The type strain is IMCC25679T (= KACC 19486T = NBRC113131T).


Subject(s)
Fresh Water/microbiology , Pedobacter/genetics , Pedobacter/isolation & purification , Bacterial Typing Techniques , Base Composition , DNA, Bacterial/genetics , Fatty Acids/analysis , Lakes , Pedobacter/chemistry , Pedobacter/classification , Phosphatidylethanolamines , Phylogeny , RNA, Ribosomal, 16S/genetics , Republic of Korea , Sequence Analysis, DNA , Sphingolipids/analysis , Vitamin K 2/analysis
13.
Environ Sci Pollut Res Int ; 24(6): 5509-5520, 2017 Feb.
Article in English | MEDLINE | ID: mdl-28028705

ABSTRACT

ABS resin wastewater is a high-temperature nitrogenous organic wastewater. It can be successfully treated with anoxic/aerobic (A/O) process. In this study, the effect of temperature on nitrogen removal and microbial community after quick temperature rise (QTR) was investigated. It was indicated that QTR from 25 to 30 °C facilitated the microbial growth and achieved a similar effluent quality as that at 25 °C. QTR from 25 to 35 °C or 40 °C resulted in higher effluent concentration of chemical oxygen demand (COD), biochemical oxygen demand (BOD5), total nitrogen (TN), and total phosphorus (TP). Illumina MiSeq pyrosequencing analysis illustrated that the richness and diversity of the bacterial community was decreased as the temperature was increased. The percentage of many functional groups was changed significantly. QTR from 25 to 40 °C also resulted in the inhibition of ammonia oxidation rate and high concentration of free ammonia, which then inhibited the growth of NOB (Nitrospira), and thus resulted in nitrite accumulation. The high temperature above 35 °C promoted the growth of a denitrifying bacterial genus, Denitratisoma, which might increase N2O production during the denitrification process.


Subject(s)
Biological Oxygen Demand Analysis , Waste Disposal, Fluid/methods , Wastewater , Acrylic Resins , Ammonia/analysis , Bioreactors/microbiology , Butadienes , Denitrification , Nitrites/analysis , Nitrogen/analysis , Phosphorus/analysis , Polystyrenes , Temperature
14.
3 Biotech ; 6(2): 178, 2016 Dec.
Article in English | MEDLINE | ID: mdl-28330250

ABSTRACT

To decipher an evolutionary lineage between two different but important bacterial groups, i.e., Pseudomonas strain (γ-Proteobacteria) and Frankia strain (actinobacteria) growing in the same ecological niche in and around of an actinorhizal plant Hippophae salicifolia D. Don, genetic diversity and comparative molecular phylogeny have been investigated using 16S rRNA gene sequences and computer-simulated and virtually directed restriction fragment length polymorphism (RFLP) through 10 restriction enzymes. Bayesian and coalescent analyses on the basis of 16S rRNA gene sequences suggested three major groups with close proximity between Pseudomonas and Frankia isolates. This result has been further validated based on the data observed through similarity coefficient value and computational RFLP. Principal component analysis and Mandel h and k statistical analysis also confirmed and strengthen the findings. Approximately 458 aligned sequence of all the taxa were used to decipher nucleotide diversity, polymorphism and gene flow between these taxa. Thus, our results suggest for a possible co-evolution or a heterologous gene transfer of distantly related microbial forms. Further, our study also advocate for the use of computer aided, virtual RFLP analysis as a cost effective and rapid identification tool.

15.
Technol Health Care ; 24 Suppl 2: S841-8, 2016 Apr 29.
Article in English | MEDLINE | ID: mdl-27177110

ABSTRACT

BACKGROUND: Despite medical progress worldwide, dental caries are still widespread. Miswak is derived from the plant Arak (Salvadora persica). It is used by Muslim peoples as a natural product for the cleansing of teeth, to ensure oral and dental hygiene. OBJECTIVE: This study was conducted to evaluate the antimicrobial effects of ethanol, methanol, and ethanol/methanol extracts of Miswak against three bacterial pathogens of the oral cavity. METHODS: The pathogens were isolated from the oral cavity of volunteers/patients and were identified on the basis of 16S rRNA gene amplification data. Sequence comparisons were made with 16S rRNA gene sequences available in the GenBank database. RESULTS: The results of sequence alignment and phylogenetic analysis identified the three pathogens as being Staphylococcus aureus strain KKU-020, Enterococcus faecalis strain KKU-021 and Klebsiella pneumoniae strain KKU-022. All Miswak extracts showed powerful antimicrobial activity against the three pathogens. The maximum zone of inhibition (40.67 ± 0.88 mm) was observed against E. faecalis KKU-021 with ethanolic extracts whilst methanolic extracts showed the minimum zone of inhibition (10.33 ± 0.88 mm) against K. pneumonia KKU-022. CONCLUSION: Based upon the significant effects of the Miswak extracts, against the oral cavity pathogens in our study, we recommend that Miswak could be used as a dental hygiene method to prevent tooth caries.


Subject(s)
Anti-Bacterial Agents/administration & dosage , Anti-Bacterial Agents/pharmacology , Enterococcus faecalis/drug effects , Klebsiella/drug effects , Mouth/microbiology , Phytotherapy , Plant Extracts/administration & dosage , Plant Extracts/pharmacology , Salvadoraceae , Staphylococcus aureus/drug effects , Databases, Genetic , Dental Caries/prevention & control , Dental Plaque/prevention & control , Genes, rRNA , Humans , Oral Hygiene
16.
mSystems ; 1(2)2016.
Article in English | MEDLINE | ID: mdl-27832214

ABSTRACT

Assignment of 16S rRNA gene sequences to operational taxonomic units (OTUs) allows microbial ecologists to overcome the inconsistencies and biases within bacterial taxonomy and provides a strategy for clustering similar sequences that do not have representatives in a reference database. I have applied the Matthews correlation coefficient to assess the ability of 15 reference-independent and -dependent clustering algorithms to assign sequences to OTUs. This metric quantifies the ability of an algorithm to reflect the relationships between sequences without the use of a reference and can be applied to any data set or method. The most consistently robust method was the average neighbor algorithm; however, for some data sets, other algorithms matched its performance.

17.
PeerJ ; 3: e1487, 2015.
Article in English | MEDLINE | ID: mdl-26664811

ABSTRACT

Background. 16S rRNA gene sequences are routinely assigned to operational taxonomic units (OTUs) that are then used to analyze complex microbial communities. A number of methods have been employed to carry out the assignment of 16S rRNA gene sequences to OTUs leading to confusion over which method is optimal. A recent study suggested that a clustering method should be selected based on its ability to generate stable OTU assignments that do not change as additional sequences are added to the dataset. In contrast, we contend that the quality of the OTU assignments, the ability of the method to properly represent the distances between the sequences, is more important. Methods. Our analysis implemented six de novo clustering algorithms including the single linkage, complete linkage, average linkage, abundance-based greedy clustering, distance-based greedy clustering, and Swarm and the open and closed-reference methods. Using two previously published datasets we used the Matthew's Correlation Coefficient (MCC) to assess the stability and quality of OTU assignments. Results. The stability of OTU assignments did not reflect the quality of the assignments. Depending on the dataset being analyzed, the average linkage and the distance and abundance-based greedy clustering methods generated OTUs that were more likely to represent the actual distances between sequences than the open and closed-reference methods. We also demonstrated that for the greedy algorithms VSEARCH produced assignments that were comparable to those produced by USEARCH making VSEARCH a viable free and open source alternative to USEARCH. Further interrogation of the reference-based methods indicated that when USEARCH or VSEARCH were used to identify the closest reference, the OTU assignments were sensitive to the order of the reference sequences because the reference sequences can be identical over the region being considered. More troubling was the observation that while both USEARCH and VSEARCH have a high level of sensitivity to detect reference sequences, the specificity of those matches was poor relative to the true best match. Discussion. Our analysis calls into question the quality and stability of OTU assignments generated by the open and closed-reference methods as implemented in current version of QIIME. This study demonstrates that de novo methods are the optimal method of assigning sequences into OTUs and that the quality of these assignments needs to be assessed for multiple methods to identify the optimal clustering method for a particular dataset.

18.
Microbiol Res ; 168(8): 485-96, 2013 Oct 01.
Article in English | MEDLINE | ID: mdl-23706760

ABSTRACT

Studies on the microbial ecology of gut microbiota in bats are limited and such information is necessary in determining the ecological significance of these hosts. Short-nosed fruit bats (Cynopterus brachyotis brachyotis) are good candidates for microbiota studies given their close association with humans in urban areas. Thus, this study explores the gut microbiota of this species from Peninsular Malaysia by means of biochemical tests and 16S rRNA gene sequences analysis. The estimation of viable bacteria present in the stomach and intestine of C. b. brachyotis ranged from 3.06×10(10) to 1.36×10(15)CFU/ml for stomach fluid and 1.92×10(10) to 6.10×10(15)CFU/ml for intestinal fluid. A total of 34 isolates from the stomach and intestine of seven C. b. brachyotis were retrieved. A total of 16 species of bacteria from eight genera (Bacillus, Enterobacter, Enterococcus, Escherichia, Klebsiella, Pantoea, Pseudomonas and Serratia) were identified, Enterobacteriaceae being the most prevalent, contributing 12 out of 16 species isolated. Most isolates from the Family Enterobacteriaceae have been reported as pathogens to humans and wildlife. With the possibility of human wildlife transmission, the findings of this study focus on the importance of bats as reservoirs of potential bacterial pathogens.


Subject(s)
Chiroptera/microbiology , Gastrointestinal Tract/microbiology , Microbiota , Animals , Bacteria/classification , Bacteria/genetics , Bacteria/isolation & purification , Bacterial Load , Phylogeny , RNA, Ribosomal, 16S/genetics
19.
Clin Exp Gastroenterol ; 5: 173-86, 2012.
Article in English | MEDLINE | ID: mdl-23049264

ABSTRACT

OBJECTIVES: The aim of this study was to define the composition of the gut bacterial flora in Norwegian patients with early stage Crohn's disease (CD). METHODS: By using a nonselective metagenomics approach, the general bacterial composition in mucosal biopsies from the ileum and the colon of five subjects, four patients with different phenotypes of CD, and one noninflammatory bowel disease control, was characterized. After partial 16S ribosomal RNA (rRNA) gene sequencing, BLAST homology searches for species identification and phylogenetic analysis were performed. RESULTS: An overall biodiversity of 106 different bacterial operational taxonomic units (OTUs) was detected in the cloned libraries. Nearly all OTUs belonged to the phylae Bacteroidetes (42% in CD, 71% in the control) or Firmicutes (42% in CD, 28% in the control), except for some OTUs that belonged to the phylum Proteobacteria (15% in CD, 0% in the control) and a few OTUs that could not be assigned to a phylum (2% in CD, 1% in the control). CONCLUSION: Based on the high incidence of inflammatory bowel disease (IBD) in Norway, this pilot study represents a relevant determination of the gut microbiota in Norwegian patients compared to previous findings in other countries. The bacterial profile of Norwegian CD patients was found to be similar to that of CD patients in other countries. The findings do not support a particular bacterial composition as a predominant causative factor for the high incidence of IBD that exists in some countries.

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