DNA was extracted from the strain of pathogen of poplar leaf blight using a modified CTAB method. ITS sequence （601bp） was initially amplified from the pathogen by using the universal primers ITSl and ITS4 （registe...DNA was extracted from the strain of pathogen of poplar leaf blight using a modified CTAB method. ITS sequence （601bp） was initially amplified from the pathogen by using the universal primers ITSl and ITS4 （registered No, DQ011257）. Comparing to the nucleotide sequences acquired from GenBank database, the strain is clustered into the homogeneity with Alternaria alternate （AY787684） and Alternaria alternate （AY354228）, with a homology of 98%, thus the strain was checked as Alternaria alternata （Fr.） Keissler. The optimal conditions for conidia germination and mycelium growth of the pathogen were tested, The optimal temperature for conidia germinating and mycelium growth is 25℃, and the optimal pH value is 6. Mycelium grows rather slowly at 10℃ and 30℃ and growth stops at above 35 ℃. Among the six culture mediums tested, PDA ＋ poplar leaf juice medium is most favorable for mycelium growth.展开更多
A sulfate reducing bacteria was isolated from mining sewage of Daqing Oilfield by Hungate anaerobic technology. Physiological-biochemical analysis showed that the strain could utilize polyacrylamide as sole carbon and...A sulfate reducing bacteria was isolated from mining sewage of Daqing Oilfield by Hungate anaerobic technology. Physiological-biochemical analysis showed that the strain could utilize polyacrylamide as sole carbon and nitrogen source. The sequence analysis of 16S rDNA illustrated that the similarity of F8 and Desulfovibrio desulfuricans (AF192153) was 99%, and the similarity sequence of dissimilatory sulfite reductase gene (DSR) cloned from the strain and Desulfovibrio desulfuricans (AF273034) was 98%. Their phylogenitic analysis was basically anastomosed, and thus temporarily named as Desulfovibrio desulfuricans F8. The DSR cloned from F8 strain was 2740 bp in length consisting of three ORF, DSRA, DSRB and DSRD as a single operon (DSRABD) regulated by the same operator. DSRA contained typical conservative box of sulfate—sulfite reducing enzyme (SiteⅠand SiteⅡ), which could bind siroheme and [Fe4S4]. DSRB retained a [Fe4S4] binding site, with an uncomplimentary structure for siroheme binding. There was no conservative box in DSRD. Sequence analysis of DSR will provide a theoretical basis for quantitative detection, metabolic pathway modification through gene engineering, and sulfate reducing bacteria (SRB) suppression.展开更多
Catalpa bungei,a fast-growing timber tree,is threatened by the lepidopteran pest Omphisa plagialis.Previous studies in our laboratory successfully generated transgenic C.bungei lines overexpressing Cry genes(Cry1Ab,Cr...Catalpa bungei,a fast-growing timber tree,is threatened by the lepidopteran pest Omphisa plagialis.Previous studies in our laboratory successfully generated transgenic C.bungei lines overexpressing Cry genes(Cry1Ab,Cry2A,and Cry9-2)that exhibited resistance to O.plagialis,but their potential impact on soil bacterial communities remains unclear.In this study,we analyzed nine transgenic C.bungei lines(three independent lines for each Cry gene)to characterize their rhizosphere bacterial communities using high-throughput sequencing of the 16S ribosomal DNA(rDNA)V4-V5 regions.A total of 628 amplicon sequence variants(ASVs)were shared among all transgenic and wild-type(WT)lines,forming a stable core microbiome dominated by Proteobacteria,Bacteroidota,Acidobacteriota,and Actinobacteriota.Alpha diversity showed no significant differences,while beta diversity revealed minor but distinct compositional shifts.Cry1Ab lines exhibited higher abundances of fast-growing taxa,particularly Proteobacteria and Bacteroidota;Cry2A lines displayed intermediate profiles,whereas Cry9-2 lines were nearly indistinguishable from WT communities.Linear discriminant analysis of the effect size revealed significant enrichment of taxa such as Burkholderiaceae and Ralstonia in the Cry1Ab rhizosphere,in contrast to the higher abundance of Chloroflexi in the WT.Functional predictions indicated consistent metabolic pathways across all treatments,suggesting strong ecological redundancy.This study demonstrates minimal impact on rhizosphere microbial communities in transgenic C.bungei plants.The Cry9-2 construct exhibited superior environmental stability,whereas the Cry1Ab construct caused only slight but ecologically acceptable shifts.These findings support the ecological safety of Bt-transgenic C.bungei and identify Cry9-2 as a particularly favorable candidate for forestry applications.This comparative evaluation of three Cry genes in a tree species provides a framework for future gene-specific biosafety assessments in woody plants.展开更多
The metagenomic DNA of disease tissue samples from four kinds of major edible fungus was extracted by CTAB method combined with DNA gel recovery kit. The genomie DNA was amplified by polymerase chain reaction using th...The metagenomic DNA of disease tissue samples from four kinds of major edible fungus was extracted by CTAB method combined with DNA gel recovery kit. The genomie DNA was amplified by polymerase chain reaction using the universal primers of 16S rDNA and 18S rDNA, and then mone, elonal sequenced after ligated and transformed, rDNA sequences of 20 positive clones were selected randomly from each pair of primers for sequence alignment. The results showed that there were two bacterial diseases and two fungul diseases in the samples, respectively.展开更多
Objective To investigate the effects of Qingguang'anⅡFormula(QGAⅡ)on the gut micro-biota of mice with chronic high intraocular pressure(IOP)model,and explore its key micro-biota for protecting the optic nerve.Me...Objective To investigate the effects of Qingguang'anⅡFormula(QGAⅡ)on the gut micro-biota of mice with chronic high intraocular pressure(IOP)model,and explore its key micro-biota for protecting the optic nerve.Methods A total of 10 specific pathogen free(SPF)grade female DBA/2J mice were random-ly divided into model group and QGAⅡgroup(n=5 for each group),while additional 5 SPF-grade female C57BL/6J mice were assigned to control group.Mice presented spontaneous high IOP and showed elevated approximately at the age of seven months.The high IOP was maintained until week 38,when gavage was initiated.Mice in control group underwent the same intragastric treatment,while those in QGAⅡgroup were gavaged with QGAⅡ(9.67 g/kg),once a day for four weeks.Retinal morphology was examined using hematoxylin and eosin(HE)staining,with the number of retinal ganglion cells(RGCs)counted.The expression level of Brn3a protein,a specific marker for RGCs,was detected by immunofluorescence,with the mean optical density(OD)measured for quantitative analysis.In addition,16S rDNA se-quencing was leveraged to analyze changes in the diversity of gut microbiota,including theirα-diversity(Chao1,Shannon,Pielou’s evenness,and observed species index)andβ-diversity.Venn diagrams and linear discriminant analysis effect size(LEfSe)analysis was employed to investigate the number of amplicon sequence variants(ASVs),the abundance of differential gut microbiota species,and the classification of species at both the phylum and genus levels within the three groups of mice.Results HE staining revealed that compared with control group,model group showed signif-icant reduction in the number of RGCs(P<0.01),with intracellular vacuolar degeneration and nuclear pyknosis.After QGAⅡtreatment,the number of RGCs was significantly in-creased compared with model group(P<0.01),with notable improvements in intracellular vacuolar degeneration.Immunofluorescence analysis showed that the mean OD of Brn3a protein was significantly decreased in model group compared with control group(P<0.01),while QGAⅡtreatment significantly elevated its expression level(P<0.01).Analysis ofα-diversity showed that after QGAⅡintervention,the Chao1,Shannon,and Pielou’s evenness indices were significantly increased(P<0.01),and the observed species index was elevated(P<0.05).β-Diversity analysis demonstrated distinct clustering among the three groups,indicating relatively low similarity in bacterial community structures.ASV clustering identi-fied a total of 14061 ASVs across all groups,with 9514 ASVs shared between model and QGAⅡgroups.At the phylum level,the abundance of Bacteroidetes was significantly decreased in model group compared with control group(P<0.01),while Firmicutes and the Firmicutes/Bacteroidetes(F/B)ratio were significantly increased(P<0.01).QGAⅡtreatment significantly reduced both Firmicutes abundance and the F/B ratio(P<0.01).At the genus level,Lactobacillus was dominant across all groups,with its abundance significantly in-creased in model group(P<0.01)and subsequently decreased following QGAⅡintervention(P<0.05).Conclusion QGAⅡrestructured the gut microbiota of DBA/2J mice with chronic high IOP,bringing changes in their diversity and abundance of components.Firmicutes,Bacteroidetes,Lactobacillus,along with their associated microorganisms,are likely critical components of the gut microbiota that contribute to the optic neuroprotective effects of QGAⅡon chronic high IOP mice.展开更多
The diagnosis of pathogenic bacteria in severe pneumonia is difficult and the prognosis is poor. Its outcome is closely related to bacterial pathogenicity and the timeliness and pertinence of antibiotic treatment. The...The diagnosis of pathogenic bacteria in severe pneumonia is difficult and the prognosis is poor. Its outcome is closely related to bacterial pathogenicity and the timeliness and pertinence of antibiotic treatment. Therefore, early diagnosis is of great significance to the prognosis of patients. Sputum examination and culture is the gold standard for the diagnosis of pathogens of severe pneumonia. However, due to the long time of bacterial culture, the early use of antibiotics, the change of bacteria species, mixed infection and other problems, the results of bacterial culture in sputum are often false negative. With the continuous application of new molecular biology techniques in clinical detection, the classification of bacteria and microorganisms has deepened from the identification of phenotypic characteristics to the classification of gene characteristics. Sequencing analysis with 16S rDNA sequencing technology has the characteristics of high sequencing flux, large amount of data obtained, short cycle, and can more comprehensively reflect the species composition of microbial community, real species distribution and abundance information. In this paper, 16S rDNA sequencing technology was used to analyze the bacterial population composition in the sputum of severe pneumonia, and to explore a new method of etiological diagnosis.展开更多
<b>Objective:</b> 120 patients with severe pneumonia who were kept in the comprehensive ICU of our hospital in 2018 were selected, and 16s rDNA sequencing was performed to analyze the composition of pathog...<b>Objective:</b> 120 patients with severe pneumonia who were kept in the comprehensive ICU of our hospital in 2018 were selected, and 16s rDNA sequencing was performed to analyze the composition of pathogenic bacteria in the sputum of severe pneumonia. <b>Methods:</b> The sputum samples of patients with severe bacterial pneumonia were collected, and the diversity of pathogens in the samples was analyzed by polymerase chain reaction (PCR) amplification and high-throughput sequencing (16s rDNA PCR-DGGE). <b>Results:</b> Sequence showed that sputum samples contained a relatively large number of species, and there were many species that were not detected by sequencing. The dominant bacteria were <i>Streptococcus, Sphingomonas, Corynebacterium, Denatobacteria, Aquobacteria, Acinetobacteria, Prevotella, Klebsiella, Pseudomonas</i>, etc. <b>Conclusion:</b> Bacteria caused by sputum of severe bacterial pneumonia are complex and diverse, which provides new methods and ideas for individualized treatment of patients with severe pneumonia.展开更多
AIM To investigate the material basis and mechanism underlying the therapeutic effect of DLC in T2DM.METHODS T2DM was triggered in rats using a high-sugar,high-fat diet alongside 35 mg/kg streptozotocin.The effect of ...AIM To investigate the material basis and mechanism underlying the therapeutic effect of DLC in T2DM.METHODS T2DM was triggered in rats using a high-sugar,high-fat diet alongside 35 mg/kg streptozotocin.The effect of DLC on the intestinal microbiota in T2DM rats was analyzed via 16S rDNA sequencing.Targeted metabolomics was conducted to evaluate the impact of DLC on the levels of nine short-chain fatty acids(SCFAs).Untargeted metabolomics examined DLC-induced alterations in fecal metabolites and associated metabolic pathways.Additionally,Spearman’s correlation analysis assessed gut microbiota and fecal metabolite relationships.RESULTS DLC significantly attenuated pathological weight loss,reduced fasting blood glucose levels,restored blood sugar homeostasis,and ameliorated dyslipidemia in T2DM rats.The 16S rDNA sequencing revealed that DLC enhanced microbial diversity and reversed intestinal dysbiosis.Targeted metabolomics indicated decreased acetic acid and propionic acid levels and increased butyric acid,isobutyric acid,and 2-methylbutyric acid levels after DLC treatment.Untargeted metabolomics revealed 57 metabolites with altered expression associated with amino acid,carbohydrate,purine,and biotin pathways.The Spearman analysis demonstrated significant links between specific gut microbiota taxa and fecal metabolites.CONCLUSION DLC may exert hypoglycemic effects by modulating intestinal flora genera,SCFA levels,and fecal metabolites.展开更多
Six strains of moderately thermophilic sulfur-oxidizing bacteria were isolated from several different typical environments in China. The identities of the isolates were confirmed by analyses of their 16S rRNA genes, a...Six strains of moderately thermophilic sulfur-oxidizing bacteria were isolated from several different typical environments in China. The identities of the isolates were confirmed by analyses of their 16S rRNA genes, and some key physiological traits. The isolates are Gram negative, rod-shaped bacteria, their optimal temperature and pH value for growth are 45-50℃ and 2.5-3.5 respectively. They are autotrophic and used elemental sulfur, sodium thiosulfate and potassium tetrathionate as electron donor, while a little glucose stimulated their growth. 16S rDNA sequences analysis reveals that the strains are phylogenetically clustered to Acidithiobacillus caldus.展开更多
Phyllosphere algae are common in tropical rainforests,forming visible biofilms or spots on plant leaf surfaces.However,knowledge of phyllosphere algal diversity and the environmental factors that drive that diversity ...Phyllosphere algae are common in tropical rainforests,forming visible biofilms or spots on plant leaf surfaces.However,knowledge of phyllosphere algal diversity and the environmental factors that drive that diversity is limited.The aim of this study is to identify the environmental factors that drive phyllosphere algal community composition and diversity in rainforests.For this purpose,we used single molecule real-time sequencing of full-length 18S rDNA to characterize the composition of phyllosphere microalgal communities growing on four host tree species(Ficus tikoua,Caryota mitis,Arenga pinnata,and Musa acuminata) common to three types of forest over four months at the Xishuangbanna Tropical Botanical Garden,Yunnan Province,China.Environmental 18S rDNA sequences revealed that the green algae orders Watanabeales and Trentepohliales were dominant in almost all algal communities and that phyllosphere algal species richness and biomass were lower in planted forest than in primeval and reserve rainforest.In addition,algal community composition differed significantly between planted forest and primeval rainforest.We also found that algal communities were affected by soluble reactive phosphorous,total nitrogen,and ammonium contents.Our findings indicate that algal community structure is significantly related to forest type and host tree species.Furthermore,this study is the first to identify environmental factors that affect phyllosphere algal communities,significantly contributing to future taxonomic research,especially for the green algae orders Watanabeales and Trentepohliales.This research also serves as an important reference for molecular diversity analysis of algae in other specific habitats,such as epiphytic algae and soil algae.展开更多
Objective:To analyze the characteristics of bacterial flora on the tongue and types of tongue coating between healthy individuals and patients with type 2 diabetes mellitus(T2DM)via detecting 16S rDNA of oral tongue c...Objective:To analyze the characteristics of bacterial flora on the tongue and types of tongue coating between healthy individuals and patients with type 2 diabetes mellitus(T2DM)via detecting 16S rDNA of oral tongue coating microbial group.Methods:A total of 42 patients with T2DM were recruited,including 6 with thin white coating,9 with mirror-like coating,27 with yellow thick coating(YTC),and 28 healthy individuals.The V4 region of 16S rDNA from the tongue coating microbiota was sequenced using the Linux ubuntu sequencing platform,and the structure of bacterial flora in the tongue was analyzed.Treeplot construction,principal coordinates analysis,redundancy analysis,and linear discriminant analysis were conducted using R software to analyze the differences of bacterial flora in the tongue coating in each group.Results:A total of 8131 different operational taxonomic units(OTUs)were obtained via sequencing,of which 719 OTU samples showed significant differences(P<0.05).Comparing OTUs with the 16S rDNA database of known species and annotation of parallel species revealed 16 species with differences at the phylum level,31 at the class level,54 at the order level,88 at the family level,and 161 at the genus level.The dominant bacteria found in patients with T2DM included Lactobacillus,Streptophyta,Chloroplast,Cyanobacteria-Chloroplast,and Bifidobacteriaceae,etc.Dominant bacteria in the control group belonged to Pasteurellales,Pasteurellaceae,Leptotrichiaceae,Lachnoanaerobaculum,and Ignavibacteria among other bacterial families.All samples were clustered into three groups,each group characterized by dominant bacteria.The order of dominant bacteria in group 1 were Prevotella>Neisseria>Veillonella>Streptococcus>Fusobacterium>Leptotrichia and so on.The order of dominant bacteria of group 2 were Neisseria>Prevotella>Fusobacterium>Streptococcus>porphyromonas and so on.The dominant bacteria of group 3 were Prevotella>Fusobacterium>Streptococcus>Neisseria>Leptotrichia>Rothia>Veillonella>porphyromonas>f_Pasteurellaceae>Capnocytophaga>Actinomyces>Alloprevotella and so on.Single factor risk estimation:with group 1 used as the reference(OR=1),the risk of carrying group 2 and group 3 microflora with T2DM increased(OR values were 4.77 and 7.78,respectively).The proportion of group 1 microflora in the control group(25.9%)was higher than that in the T2DM group(5.4%)(χ2=3.873,P=0.049).Compared with group 1 and group 2,group 3 had a significantly dominant microflora structure in YTC group(χ2=7.120,P=0.008).RDA analysis indicated that Neisseria,Fusobacterium,and Prevotella were associated with HbA1c values>10 mmol/L,whereas Rothia,Streptococcus,and Veillonella were more correlated with HbA1c≤10 mmol/L.Conclusion:The incidence of T2DM is related to the imbalance of oral microflora in the human body.The tongue coating flora structure may influence the formation of different tongue coating types in this metabolic condition.Additionally,flora structures of Prevotella,Neisseria,and Fusobacterium were associated with elevated HbA1c.展开更多
The yeast strain （Y18） was isolated from a soil sample collected from Fildes Peninsula, Antarctica. The strain is a psychrophilic yeast with optimum and maximum growth temperatures of 10 ℃ and 18 ℃, respectively. ...The yeast strain （Y18） was isolated from a soil sample collected from Fildes Peninsula, Antarctica. The strain is a psychrophilic yeast with optimum and maximum growth temperatures of 10 ℃ and 18 ℃, respectively. Teliospores were formed after 7 d on malt agar, when the germination of teliospores was observed. Both inositol and D-glucuronate were assimilated. Positive results of the DBB （diazonium blue B） color reaction, urease test, and starch formation were observed. The major CoQ is Qs. All results indicated that Y18 belongs to the genes of Mrakia. The 18S rDNA sequence analyses showed that Y18 is closely related to Mrakiafrigida. DNA-DNA relatedness study, and some biochemistry characteristics indicated that Y18 represents a new species for which Mrakia psychrophila sp. nov. is proposed.展开更多
A slight halophilic heterotrophic nitrobacteria named gs1 was separated from the matured activated sludge.According to the morphological observation,physiological biochemical tests and sequence analysis of the 16S rDN...A slight halophilic heterotrophic nitrobacteria named gs1 was separated from the matured activated sludge.According to the morphological observation,physiological biochemical tests and sequence analysis of the 16S rDNA,strain gs1 was identified to be as Pseudomonas sp.Sodium acetate and ammonium chloride were used as carbon and nitrogen sources,respectively,to investi-gate the characteristics of the bacterium.When cultured for 24 h under aerobic conditions,with the removal rates of the NH4+-N and COD being 82.2%and 74.73%,respectively,strain gs1 will have a nitrification function of producing NO2--N.When cultured for 24 h under aerobic conditions in nitrite medium,the removal rate of the NO2--N became 100%,and when cultured for 24 h under aerobic conditions in nitrate medium,the removal rate of the NO3--N became 97%.The result shows that this strain functions for either nitrification or denitrification,i.e.,it can complete the full process of biological deoxidation.展开更多
The anaerobic process of biohydrogen production was developed recently. The isolation and identification of biohydrogen producing anaerobic bacteria with high evolution rate and yield is an important foundation of the...The anaerobic process of biohydrogen production was developed recently. The isolation and identification of biohydrogen producing anaerobic bacteria with high evolution rate and yield is an important foundation of the fermented biohydrogen production process through which anaerobic bacteria digest organic wastewater. By considering physiological and biochemical traits, morphological characteristics and a 16S rDNA sequence, the isolated Rennanqilyf33 is shown to be a new species.展开更多
While Ramularia leaf blight of cotton caused by Ramularia areola is of top most importance for Brazil, information is lacking regarding the survival mechanism of this pathogen during the cotton-free period. The teleom...While Ramularia leaf blight of cotton caused by Ramularia areola is of top most importance for Brazil, information is lacking regarding the survival mechanism of this pathogen during the cotton-free period. The teleomorph of R. areola is expected to belong to the genus Mycosphaerella. In the present study attempts were made to verify occurrence of this teleomorph in the State of Mato Grosso, Brazil. Decaying cotton leaves were collected two months after harvest of 2014 from 44 commercial and experimental fields where aerial fungicidal applications were made or not during the crop cycle to control the Ramularia leaf blight. Examination of the decaying cotton leaves revealed presence of abundant sclerotia, spermagonia and ascoma of Mycosphaerella sp. intermingled with each other during the cotton-free period in most of the leaf samples. Mono-ascospore isolations were obtained from the ascoma and considering their cultural, morphological, pathological and DNA sequence analysis they were identified as Mycosphaerella areola. M. areola and R. areola isolates produced similar symptoms under glasshouse inoculations. Reisolation of the pathogen from the symptoms produced by M. areola isolates yielded R. areola. Some cotton leaves showing such symptoms were kept on the soil surface on plastic trays for two months under natural field condition. After this period the decaying leaves showed abundant perithecia identical to their original M. areola. ITS rDNA sequence analyses revealed identical sequences from M. areola and R. areola isolates. Occurrence and the viability of the perfect stage M. areola during the cotton-free period on the left-over stubble from one season to another were interpreted as the survival mechanism of the pathogen and were considered responsible for the Ramularia blight epidemics in the State of Mato Grosso. Disease management practices for the State of Mato Grosso are discussed. This is the first report about the occurrence of the M. areola in Brazil.展开更多
BACKGROUND This study aimed to identify characteristic gut genera in obese and normal-weight children(8-12 years old)using 16S rDNA sequencing.The research aimed to provide insights for mechanistic studies and prevent...BACKGROUND This study aimed to identify characteristic gut genera in obese and normal-weight children(8-12 years old)using 16S rDNA sequencing.The research aimed to provide insights for mechanistic studies and prevention strategies for childhood obesity.Thirty normal-weight and thirty age-and sex-matched obese children were included.Questionnaires and body measurements were collected,and fecal samples underwent 16S rDNA sequencing.Significant differences in body mass index(BMI)and body-fat percentage were observed between the groups.Analysis of gut microbiota diversity revealed lowerα-diversity in obese children.Differences in gut microbiota composition were found between the two groups.Prevotella and Firmicutes were more abundant in the obese group,while Bacteroides and Sanguibacteroides were more prevalent in the control group.AIM To identify the characteristic gut genera in obese and normal-weight children(8-12-year-old)using 16S rDNA sequencing,and provide a basis for subsequent mechanistic studies and prevention strategies for childhood obesity.METHODS Thirty each normal-weight,1:1 matched for age and sex,and obese children,with an obese status from 2020 to 2022,were included in the control and obese groups,respectively.Basic information was collected through questionnaires and body measurements were obtained from both obese and normal-weight children.Fecal samples were collected from both groups and subjected to 16S rDNA sequencing using an Illumina MiSeq sequencing platform for gut microbiota diversity analysis.RESULTS Significant differences in BMI and body-fat percentage were observed between the two groups.The Ace and Chao1 indices were significantly lower in the obese group than those in the control group,whereas differences were not significant in the Shannon and Simpson indices.Kruskal-Wallis tests indicated significant differences in unweighted and weighted UniFrac distances between the gut microbiota of normal-weight and obese children(P<0.01),suggesting substantial disparities in both the species and quantity of gut microbiota between the two groups.Prevotella,Firmicutes,Bacteroides,and Sanguibacteroides were more abundant in the obese and control groups,respectively.Heatmap results demonstrated significant differences in the gut microbiota composition between obese and normal-weight children.CONCLUSION Obese children exhibited lowerα-diversity in their gut microbiota than did the normal-weight children.Significant differences were observed in the composition of gut microbiota between obese and normal-weight children.展开更多
Wantong Jingu Tablet(WJT),a mixture of traditional Chinese medicine,was reported to relieve the symptoms of rheumatoid arthritis(RA),but its pharmacological mechanism was not completely understood.The aim of this stud...Wantong Jingu Tablet(WJT),a mixture of traditional Chinese medicine,was reported to relieve the symptoms of rheumatoid arthritis(RA),but its pharmacological mechanism was not completely understood.The aim of this study was to investigate the therapeutic mechanisms of WJT for RA in vivo.The effects of WJT on joint pathology,as well as the levels of Bax,Bcl-2,caspase-3,cleaved-caspase-3,ERK1/2,pERK1/2,TNF-α,IL-1β,and IL-6 were measured using collagen-induced arthritis(CIA)rats.The intestinal flora composition and the metabolites alteration were analyzed by 16S rDNA sequencing and metabolomics method,respectively.We found that WJT ameliorated the severity of the CIA rats which might be mediated by inducing apoptosis,inactivating the MEK/ERK signals and reducing the production of pro-inflammatory cytokines.WJT,in part,relieved the gut microbiota dysbiosis,especially bacterial phylum Bacteroidetes,Tenericutes and Deferribacteres,as well as bacterial genus Vibrio,Macrococcus and Vagococcus.3’-N-debenzoyl-2’-deoxytaxol,tubulysin B,and magnoline were significantly associated with the specific genera.We identified serotonin,glutathione disulfide,N-acetylneuraminic acid,naphthalene and thromboxane B2 as targeted molecules via metabolomics.Our findings contributed to the understanding of RA pathogenesis,and WJT played essential roles in gut microbiota health and metabolite modulation in the CIA rats.展开更多
Dinoflagellates are unicellular eukaryotic protists that dominate in all coastal waters, and are also present in oceanic waters. Despite the central importance of dinoflagellates in global primary production, the rela...Dinoflagellates are unicellular eukaryotic protists that dominate in all coastal waters, and are also present in oceanic waters. Despite the central importance of dinoflagellates in global primary production, the relationship between dinoflagellates and bacteria are still poorly understood. In order to understand the ecological interaction between bacterial and dinoflagellates communities, denaturing gradient gel electrophoresis (DGGE) and SSU rRNA sequencing were applied to monitoring the population dynamics of bacteria and dinoflagellates from the onset to disappearance of a dinoflagellates bloom occurred in Baltimore Inner Harbor, from April 15 to 24,2002. Although Prorocentrum minimum was the major bloom forming species under the light microscopy, DGGE method with dinoflagellate specific primers demonstrated that Prorocentrum micans, Gymnodinium galatheanum and Gyrodinium uncatenum were also present during the bloom. Population shifts among the minor dinoflagellate groups were observed. DGGE of PCR-amplified 16S rRNA gene fragments indicated that cyanobacteria, α, β, γ-proteobacteria, Flavobacterium Bacteroides-Cytophaga (FBC), and Planctomcetes were the major components of bacterial assemblages during the bloom. DGGE analysis showed that Cytophagales and α-proteobacteria played important roles at different stages of dinoflagellates bloom. DGGE can be used as a rapid tool to simultaneously monitor population dynamics of both bacterial and dinoflagellates communities in aquatic environments, which is demonstrated here.展开更多
We investigated the diversity and composition of microflora in feces of Lycopus lucidus Turcz.-fed mice.In addition,we evaluated the production of major cytokines(Interleukin-6 and-10)which are related to inflammation...We investigated the diversity and composition of microflora in feces of Lycopus lucidus Turcz.-fed mice.In addition,we evaluated the production of major cytokines(Interleukin-6 and-10)which are related to inflammation and fatty acid composition of several tissues.16S ribosomal DNA sequencing-based microbiome taxonomic profiling analysis was performed utilizing the EzBioCloud data base.Male mice fed on L.lucidus showed a significantly reduced number of lactic acid bacteria and coliform in the feces compared with the control group(p<0.05).16S rDNA sequencing analysis of fecal samples showed that L.lucidus supplementation decreased the community of harmful microflora(Enterobacteriaceae including Escherichia coli and Bacteroides sp.)in feces compared with the control group(p<0.05).There were no significant differences in mRNA expression of cytokine IL-6 and IL-10 between the control and L.lucidus fed groups.The fecal fatty acid composition in the L.lucidus group had percentages of 4:0,6:0,8:0 and 10:0 in the intestine but those short chain fatty acids were not detected in the control group.Our results showed that L.lucidus supplementation influenced gut environment by decreasing harmful microflora and increased the percentages of several short fatty acids.展开更多
Objective:To explore the structural and functional characteristics of microbiota in oropharynx of subhealthy children with gastrointestinal heat retention syndrome(GHRS)differentiated by traditional Chinese medicine,a...Objective:To explore the structural and functional characteristics of microbiota in oropharynx of subhealthy children with gastrointestinal heat retention syndrome(GHRS)differentiated by traditional Chinese medicine,and screen the biometric operational taxonomic units(OTUs)to assist the clinical diagnosis.Methods:We recruited children according to the“GHRS diagnostic scale”,collected their oropharyngeal swabs,and sequenced the 16 SrDNA V4 region.We described the bacterial structure with alpha-indexes,beta-distances,and relative abundances;moreover,we screened the differential genera/OTUs with Wilcoxon rank-sum test,Metagenome Seq analysis,and linear discriminant analysis effect size(LEf Se)analysis,in which biometric OTUs were selected to construct the receiver operating characteristic curve to verify the diagnostic value.The bacterial function was predicted with Kyoto Encyclopedia of Genes and Genomes pathways according to 16S rDNA gene by using Phylogenetic Investigation of Communities by Reconstruction of Unobserved States.Results:The study population was composed of 10 children with GHRS and 10 healthy control children.GHRS children were more likely to overeat(gluttony,P=.033).Alpha-indexes,such as Sobs,abundancebased coverage estimator,Bootstrap,and Qstat,were significantly higher in the GHRS group,while betadistances did not exhibit any significant intergroup differences.There were 9 differently distributed nonpredominant genera between the groups in Wilcoxon rank-sum test,as well as 13 non-predominant genera in Metagenome Seq analysis and 3 non-predominant OTUs in LEfSe analysis.OTU44 and OTU196 were used to construct the receiver operating characteristic curve,and the area under curve was 0.92.Predicted functions showed that pathways related to oxidative phosphorylation and carbon metabolism were enriched in healthy control samples,while the pathway related to renin secretion was remarkably enriched in GHRS samples.Conclusion:Unique oropharyngeal microbial structure and function were identified in GHRS children.OTU44 and OTU196 were specific OTUs,which could be used as biomarkers of GHRS to assist clinical diagnosis.展开更多
基金This paper was supported by National Natural Science Foundation of China （No.30271083）.
文摘DNA was extracted from the strain of pathogen of poplar leaf blight using a modified CTAB method. ITS sequence （601bp） was initially amplified from the pathogen by using the universal primers ITSl and ITS4 （registered No, DQ011257）. Comparing to the nucleotide sequences acquired from GenBank database, the strain is clustered into the homogeneity with Alternaria alternate （AY787684） and Alternaria alternate （AY354228）, with a homology of 98%, thus the strain was checked as Alternaria alternata （Fr.） Keissler. The optimal conditions for conidia germination and mycelium growth of the pathogen were tested, The optimal temperature for conidia germinating and mycelium growth is 25℃, and the optimal pH value is 6. Mycelium grows rather slowly at 10℃ and 30℃ and growth stops at above 35 ℃. Among the six culture mediums tested, PDA ＋ poplar leaf juice medium is most favorable for mycelium growth.
基金Sponsored by the National Basic Research and Development (973) Program of China(Grant No.2004CB418505)
文摘A sulfate reducing bacteria was isolated from mining sewage of Daqing Oilfield by Hungate anaerobic technology. Physiological-biochemical analysis showed that the strain could utilize polyacrylamide as sole carbon and nitrogen source. The sequence analysis of 16S rDNA illustrated that the similarity of F8 and Desulfovibrio desulfuricans (AF192153) was 99%, and the similarity sequence of dissimilatory sulfite reductase gene (DSR) cloned from the strain and Desulfovibrio desulfuricans (AF273034) was 98%. Their phylogenitic analysis was basically anastomosed, and thus temporarily named as Desulfovibrio desulfuricans F8. The DSR cloned from F8 strain was 2740 bp in length consisting of three ORF, DSRA, DSRB and DSRD as a single operon (DSRABD) regulated by the same operator. DSRA contained typical conservative box of sulfate—sulfite reducing enzyme (SiteⅠand SiteⅡ), which could bind siroheme and [Fe4S4]. DSRB retained a [Fe4S4] binding site, with an uncomplimentary structure for siroheme binding. There was no conservative box in DSRD. Sequence analysis of DSR will provide a theoretical basis for quantitative detection, metabolic pathway modification through gene engineering, and sulfate reducing bacteria (SRB) suppression.
基金funded by the Chinese Academy of Forestry-Special funds for basic scientific research service expenses of the central level public welfare research institutes(Grant No.CAFYBB2020QD001)the National Natural Science Foundation of China(Grant Nos.32101550,32271917)+1 种基金Jiangsu Agricultural Science and Technology Innovation Fund(Grant No.CX(24)3052)National Forestry and Grassland Administration’s Center for Science and Technology Development Projects(Grant No.KJZXSA202202).
文摘Catalpa bungei,a fast-growing timber tree,is threatened by the lepidopteran pest Omphisa plagialis.Previous studies in our laboratory successfully generated transgenic C.bungei lines overexpressing Cry genes(Cry1Ab,Cry2A,and Cry9-2)that exhibited resistance to O.plagialis,but their potential impact on soil bacterial communities remains unclear.In this study,we analyzed nine transgenic C.bungei lines(three independent lines for each Cry gene)to characterize their rhizosphere bacterial communities using high-throughput sequencing of the 16S ribosomal DNA(rDNA)V4-V5 regions.A total of 628 amplicon sequence variants(ASVs)were shared among all transgenic and wild-type(WT)lines,forming a stable core microbiome dominated by Proteobacteria,Bacteroidota,Acidobacteriota,and Actinobacteriota.Alpha diversity showed no significant differences,while beta diversity revealed minor but distinct compositional shifts.Cry1Ab lines exhibited higher abundances of fast-growing taxa,particularly Proteobacteria and Bacteroidota;Cry2A lines displayed intermediate profiles,whereas Cry9-2 lines were nearly indistinguishable from WT communities.Linear discriminant analysis of the effect size revealed significant enrichment of taxa such as Burkholderiaceae and Ralstonia in the Cry1Ab rhizosphere,in contrast to the higher abundance of Chloroflexi in the WT.Functional predictions indicated consistent metabolic pathways across all treatments,suggesting strong ecological redundancy.This study demonstrates minimal impact on rhizosphere microbial communities in transgenic C.bungei plants.The Cry9-2 construct exhibited superior environmental stability,whereas the Cry1Ab construct caused only slight but ecologically acceptable shifts.These findings support the ecological safety of Bt-transgenic C.bungei and identify Cry9-2 as a particularly favorable candidate for forestry applications.This comparative evaluation of three Cry genes in a tree species provides a framework for future gene-specific biosafety assessments in woody plants.
基金Supported by Project for State Edible Fungus Industrial Technology System Construction(CARS-24)Agricultural Improved Variety Project of Shandong Province(2012LZ006-04)Science and Technology Development Program of Shandong Province
文摘The metagenomic DNA of disease tissue samples from four kinds of major edible fungus was extracted by CTAB method combined with DNA gel recovery kit. The genomie DNA was amplified by polymerase chain reaction using the universal primers of 16S rDNA and 18S rDNA, and then mone, elonal sequenced after ligated and transformed, rDNA sequences of 20 positive clones were selected randomly from each pair of primers for sequence alignment. The results showed that there were two bacterial diseases and two fungul diseases in the samples, respectively.
基金National Natural Science Foundation of China(81904260)Scientific Research Project of Hunan Province Department of Education(22B0398)Scientific Research Project of Traditional Chinese Medicine of Hunan Province(D2022045).
文摘Objective To investigate the effects of Qingguang'anⅡFormula(QGAⅡ)on the gut micro-biota of mice with chronic high intraocular pressure(IOP)model,and explore its key micro-biota for protecting the optic nerve.Methods A total of 10 specific pathogen free(SPF)grade female DBA/2J mice were random-ly divided into model group and QGAⅡgroup(n=5 for each group),while additional 5 SPF-grade female C57BL/6J mice were assigned to control group.Mice presented spontaneous high IOP and showed elevated approximately at the age of seven months.The high IOP was maintained until week 38,when gavage was initiated.Mice in control group underwent the same intragastric treatment,while those in QGAⅡgroup were gavaged with QGAⅡ(9.67 g/kg),once a day for four weeks.Retinal morphology was examined using hematoxylin and eosin(HE)staining,with the number of retinal ganglion cells(RGCs)counted.The expression level of Brn3a protein,a specific marker for RGCs,was detected by immunofluorescence,with the mean optical density(OD)measured for quantitative analysis.In addition,16S rDNA se-quencing was leveraged to analyze changes in the diversity of gut microbiota,including theirα-diversity(Chao1,Shannon,Pielou’s evenness,and observed species index)andβ-diversity.Venn diagrams and linear discriminant analysis effect size(LEfSe)analysis was employed to investigate the number of amplicon sequence variants(ASVs),the abundance of differential gut microbiota species,and the classification of species at both the phylum and genus levels within the three groups of mice.Results HE staining revealed that compared with control group,model group showed signif-icant reduction in the number of RGCs(P<0.01),with intracellular vacuolar degeneration and nuclear pyknosis.After QGAⅡtreatment,the number of RGCs was significantly in-creased compared with model group(P<0.01),with notable improvements in intracellular vacuolar degeneration.Immunofluorescence analysis showed that the mean OD of Brn3a protein was significantly decreased in model group compared with control group(P<0.01),while QGAⅡtreatment significantly elevated its expression level(P<0.01).Analysis ofα-diversity showed that after QGAⅡintervention,the Chao1,Shannon,and Pielou’s evenness indices were significantly increased(P<0.01),and the observed species index was elevated(P<0.05).β-Diversity analysis demonstrated distinct clustering among the three groups,indicating relatively low similarity in bacterial community structures.ASV clustering identi-fied a total of 14061 ASVs across all groups,with 9514 ASVs shared between model and QGAⅡgroups.At the phylum level,the abundance of Bacteroidetes was significantly decreased in model group compared with control group(P<0.01),while Firmicutes and the Firmicutes/Bacteroidetes(F/B)ratio were significantly increased(P<0.01).QGAⅡtreatment significantly reduced both Firmicutes abundance and the F/B ratio(P<0.01).At the genus level,Lactobacillus was dominant across all groups,with its abundance significantly in-creased in model group(P<0.01)and subsequently decreased following QGAⅡintervention(P<0.05).Conclusion QGAⅡrestructured the gut microbiota of DBA/2J mice with chronic high IOP,bringing changes in their diversity and abundance of components.Firmicutes,Bacteroidetes,Lactobacillus,along with their associated microorganisms,are likely critical components of the gut microbiota that contribute to the optic neuroprotective effects of QGAⅡon chronic high IOP mice.
文摘The diagnosis of pathogenic bacteria in severe pneumonia is difficult and the prognosis is poor. Its outcome is closely related to bacterial pathogenicity and the timeliness and pertinence of antibiotic treatment. Therefore, early diagnosis is of great significance to the prognosis of patients. Sputum examination and culture is the gold standard for the diagnosis of pathogens of severe pneumonia. However, due to the long time of bacterial culture, the early use of antibiotics, the change of bacteria species, mixed infection and other problems, the results of bacterial culture in sputum are often false negative. With the continuous application of new molecular biology techniques in clinical detection, the classification of bacteria and microorganisms has deepened from the identification of phenotypic characteristics to the classification of gene characteristics. Sequencing analysis with 16S rDNA sequencing technology has the characteristics of high sequencing flux, large amount of data obtained, short cycle, and can more comprehensively reflect the species composition of microbial community, real species distribution and abundance information. In this paper, 16S rDNA sequencing technology was used to analyze the bacterial population composition in the sputum of severe pneumonia, and to explore a new method of etiological diagnosis.
文摘<b>Objective:</b> 120 patients with severe pneumonia who were kept in the comprehensive ICU of our hospital in 2018 were selected, and 16s rDNA sequencing was performed to analyze the composition of pathogenic bacteria in the sputum of severe pneumonia. <b>Methods:</b> The sputum samples of patients with severe bacterial pneumonia were collected, and the diversity of pathogens in the samples was analyzed by polymerase chain reaction (PCR) amplification and high-throughput sequencing (16s rDNA PCR-DGGE). <b>Results:</b> Sequence showed that sputum samples contained a relatively large number of species, and there were many species that were not detected by sequencing. The dominant bacteria were <i>Streptococcus, Sphingomonas, Corynebacterium, Denatobacteria, Aquobacteria, Acinetobacteria, Prevotella, Klebsiella, Pseudomonas</i>, etc. <b>Conclusion:</b> Bacteria caused by sputum of severe bacterial pneumonia are complex and diverse, which provides new methods and ideas for individualized treatment of patients with severe pneumonia.
基金Supported by the National Natural Science Foundation of China,No.82160771NATCM's Project of High-Level Construction of Key TCM Disciplines:Traditional Medicine of Chinese Minority(Zhuang Medicine),No.zyyzdxk-2023165+7 种基金Guangxi One Thousand Young and Middle-Aged College and University Backbones Teachers Cultivation Program,No.[2019]5Guangxi Traditional Chinese Medicine Multidisciplinary Cross Innovation Team Project,No.GZKJ2309Guangxi Key R&D Plan Project,No.AB21196016Guangxi Key Discipline of Traditional Chinese Medicine Zhuang Pharmacy,No.GZXK-Z-20-64The First-Class Subject of Traditional Chinese Medicine(Ethnic Pharmacy)in Guangxi,No.[2018]12Guangxi Science and Technology Base and Talent Special Project,No.AD20238058 and No.AD21238031the Third Batch of Cultivating High-level Talent Teams in the“Qi Huang Project”of the Guangxi University of Chinese Medicine,No.202406and Huang Danian Style Teacher Team From Universities in Guangxi Zhuang Autonomous Region“Traditional Chinese Medicine Inheritance and Innovation Teacher Team”,No.[2023]31.
文摘AIM To investigate the material basis and mechanism underlying the therapeutic effect of DLC in T2DM.METHODS T2DM was triggered in rats using a high-sugar,high-fat diet alongside 35 mg/kg streptozotocin.The effect of DLC on the intestinal microbiota in T2DM rats was analyzed via 16S rDNA sequencing.Targeted metabolomics was conducted to evaluate the impact of DLC on the levels of nine short-chain fatty acids(SCFAs).Untargeted metabolomics examined DLC-induced alterations in fecal metabolites and associated metabolic pathways.Additionally,Spearman’s correlation analysis assessed gut microbiota and fecal metabolite relationships.RESULTS DLC significantly attenuated pathological weight loss,reduced fasting blood glucose levels,restored blood sugar homeostasis,and ameliorated dyslipidemia in T2DM rats.The 16S rDNA sequencing revealed that DLC enhanced microbial diversity and reversed intestinal dysbiosis.Targeted metabolomics indicated decreased acetic acid and propionic acid levels and increased butyric acid,isobutyric acid,and 2-methylbutyric acid levels after DLC treatment.Untargeted metabolomics revealed 57 metabolites with altered expression associated with amino acid,carbohydrate,purine,and biotin pathways.The Spearman analysis demonstrated significant links between specific gut microbiota taxa and fecal metabolites.CONCLUSION DLC may exert hypoglycemic effects by modulating intestinal flora genera,SCFA levels,and fecal metabolites.
基金Project (50321402) supported by the National Natural Science Foundation of China Project(2004CB619204) supported by the State Basic Research Development Program of China Project (DY105-02-04-05) supported by the China Ocean Mineral Resources Research and Development Association
文摘Six strains of moderately thermophilic sulfur-oxidizing bacteria were isolated from several different typical environments in China. The identities of the isolates were confirmed by analyses of their 16S rRNA genes, and some key physiological traits. The isolates are Gram negative, rod-shaped bacteria, their optimal temperature and pH value for growth are 45-50℃ and 2.5-3.5 respectively. They are autotrophic and used elemental sulfur, sodium thiosulfate and potassium tetrathionate as electron donor, while a little glucose stimulated their growth. 16S rDNA sequences analysis reveals that the strains are phylogenetically clustered to Acidithiobacillus caldus.
基金supported by the National Natural Science Foundation of China (Grant no.31870189 and 32000168)。
文摘Phyllosphere algae are common in tropical rainforests,forming visible biofilms or spots on plant leaf surfaces.However,knowledge of phyllosphere algal diversity and the environmental factors that drive that diversity is limited.The aim of this study is to identify the environmental factors that drive phyllosphere algal community composition and diversity in rainforests.For this purpose,we used single molecule real-time sequencing of full-length 18S rDNA to characterize the composition of phyllosphere microalgal communities growing on four host tree species(Ficus tikoua,Caryota mitis,Arenga pinnata,and Musa acuminata) common to three types of forest over four months at the Xishuangbanna Tropical Botanical Garden,Yunnan Province,China.Environmental 18S rDNA sequences revealed that the green algae orders Watanabeales and Trentepohliales were dominant in almost all algal communities and that phyllosphere algal species richness and biomass were lower in planted forest than in primeval and reserve rainforest.In addition,algal community composition differed significantly between planted forest and primeval rainforest.We also found that algal communities were affected by soluble reactive phosphorous,total nitrogen,and ammonium contents.Our findings indicate that algal community structure is significantly related to forest type and host tree species.Furthermore,this study is the first to identify environmental factors that affect phyllosphere algal communities,significantly contributing to future taxonomic research,especially for the green algae orders Watanabeales and Trentepohliales.This research also serves as an important reference for molecular diversity analysis of algae in other specific habitats,such as epiphytic algae and soil algae.
文摘Objective:To analyze the characteristics of bacterial flora on the tongue and types of tongue coating between healthy individuals and patients with type 2 diabetes mellitus(T2DM)via detecting 16S rDNA of oral tongue coating microbial group.Methods:A total of 42 patients with T2DM were recruited,including 6 with thin white coating,9 with mirror-like coating,27 with yellow thick coating(YTC),and 28 healthy individuals.The V4 region of 16S rDNA from the tongue coating microbiota was sequenced using the Linux ubuntu sequencing platform,and the structure of bacterial flora in the tongue was analyzed.Treeplot construction,principal coordinates analysis,redundancy analysis,and linear discriminant analysis were conducted using R software to analyze the differences of bacterial flora in the tongue coating in each group.Results:A total of 8131 different operational taxonomic units(OTUs)were obtained via sequencing,of which 719 OTU samples showed significant differences(P<0.05).Comparing OTUs with the 16S rDNA database of known species and annotation of parallel species revealed 16 species with differences at the phylum level,31 at the class level,54 at the order level,88 at the family level,and 161 at the genus level.The dominant bacteria found in patients with T2DM included Lactobacillus,Streptophyta,Chloroplast,Cyanobacteria-Chloroplast,and Bifidobacteriaceae,etc.Dominant bacteria in the control group belonged to Pasteurellales,Pasteurellaceae,Leptotrichiaceae,Lachnoanaerobaculum,and Ignavibacteria among other bacterial families.All samples were clustered into three groups,each group characterized by dominant bacteria.The order of dominant bacteria in group 1 were Prevotella>Neisseria>Veillonella>Streptococcus>Fusobacterium>Leptotrichia and so on.The order of dominant bacteria of group 2 were Neisseria>Prevotella>Fusobacterium>Streptococcus>porphyromonas and so on.The dominant bacteria of group 3 were Prevotella>Fusobacterium>Streptococcus>Neisseria>Leptotrichia>Rothia>Veillonella>porphyromonas>f_Pasteurellaceae>Capnocytophaga>Actinomyces>Alloprevotella and so on.Single factor risk estimation:with group 1 used as the reference(OR=1),the risk of carrying group 2 and group 3 microflora with T2DM increased(OR values were 4.77 and 7.78,respectively).The proportion of group 1 microflora in the control group(25.9%)was higher than that in the T2DM group(5.4%)(χ2=3.873,P=0.049).Compared with group 1 and group 2,group 3 had a significantly dominant microflora structure in YTC group(χ2=7.120,P=0.008).RDA analysis indicated that Neisseria,Fusobacterium,and Prevotella were associated with HbA1c values>10 mmol/L,whereas Rothia,Streptococcus,and Veillonella were more correlated with HbA1c≤10 mmol/L.Conclusion:The incidence of T2DM is related to the imbalance of oral microflora in the human body.The tongue coating flora structure may influence the formation of different tongue coating types in this metabolic condition.Additionally,flora structures of Prevotella,Neisseria,and Fusobacterium were associated with elevated HbA1c.
基金Project (No. 30170001) supported by the National Natural Science Foundation of China
文摘The yeast strain （Y18） was isolated from a soil sample collected from Fildes Peninsula, Antarctica. The strain is a psychrophilic yeast with optimum and maximum growth temperatures of 10 ℃ and 18 ℃, respectively. Teliospores were formed after 7 d on malt agar, when the germination of teliospores was observed. Both inositol and D-glucuronate were assimilated. Positive results of the DBB （diazonium blue B） color reaction, urease test, and starch formation were observed. The major CoQ is Qs. All results indicated that Y18 belongs to the genes of Mrakia. The 18S rDNA sequence analyses showed that Y18 is closely related to Mrakiafrigida. DNA-DNA relatedness study, and some biochemistry characteristics indicated that Y18 represents a new species for which Mrakia psychrophila sp. nov. is proposed.
基金Supported by the National Natural Science Foundation of China(50678085,50878107)the Innovative Programs Foundation of Graduate Education in Shandong Province,China(SDYY07091)the Cultivate Pro-ject of Excellent Graduate Students’Thesis of Qingdao University,China(YSPY2009014)
文摘A slight halophilic heterotrophic nitrobacteria named gs1 was separated from the matured activated sludge.According to the morphological observation,physiological biochemical tests and sequence analysis of the 16S rDNA,strain gs1 was identified to be as Pseudomonas sp.Sodium acetate and ammonium chloride were used as carbon and nitrogen sources,respectively,to investi-gate the characteristics of the bacterium.When cultured for 24 h under aerobic conditions,with the removal rates of the NH4+-N and COD being 82.2%and 74.73%,respectively,strain gs1 will have a nitrification function of producing NO2--N.When cultured for 24 h under aerobic conditions in nitrite medium,the removal rate of the NO2--N became 100%,and when cultured for 24 h under aerobic conditions in nitrate medium,the removal rate of the NO3--N became 97%.The result shows that this strain functions for either nitrification or denitrification,i.e.,it can complete the full process of biological deoxidation.
文摘The anaerobic process of biohydrogen production was developed recently. The isolation and identification of biohydrogen producing anaerobic bacteria with high evolution rate and yield is an important foundation of the fermented biohydrogen production process through which anaerobic bacteria digest organic wastewater. By considering physiological and biochemical traits, morphological characteristics and a 16S rDNA sequence, the isolated Rennanqilyf33 is shown to be a new species.
文摘While Ramularia leaf blight of cotton caused by Ramularia areola is of top most importance for Brazil, information is lacking regarding the survival mechanism of this pathogen during the cotton-free period. The teleomorph of R. areola is expected to belong to the genus Mycosphaerella. In the present study attempts were made to verify occurrence of this teleomorph in the State of Mato Grosso, Brazil. Decaying cotton leaves were collected two months after harvest of 2014 from 44 commercial and experimental fields where aerial fungicidal applications were made or not during the crop cycle to control the Ramularia leaf blight. Examination of the decaying cotton leaves revealed presence of abundant sclerotia, spermagonia and ascoma of Mycosphaerella sp. intermingled with each other during the cotton-free period in most of the leaf samples. Mono-ascospore isolations were obtained from the ascoma and considering their cultural, morphological, pathological and DNA sequence analysis they were identified as Mycosphaerella areola. M. areola and R. areola isolates produced similar symptoms under glasshouse inoculations. Reisolation of the pathogen from the symptoms produced by M. areola isolates yielded R. areola. Some cotton leaves showing such symptoms were kept on the soil surface on plastic trays for two months under natural field condition. After this period the decaying leaves showed abundant perithecia identical to their original M. areola. ITS rDNA sequence analyses revealed identical sequences from M. areola and R. areola isolates. Occurrence and the viability of the perfect stage M. areola during the cotton-free period on the left-over stubble from one season to another were interpreted as the survival mechanism of the pathogen and were considered responsible for the Ramularia blight epidemics in the State of Mato Grosso. Disease management practices for the State of Mato Grosso are discussed. This is the first report about the occurrence of the M. areola in Brazil.
文摘BACKGROUND This study aimed to identify characteristic gut genera in obese and normal-weight children(8-12 years old)using 16S rDNA sequencing.The research aimed to provide insights for mechanistic studies and prevention strategies for childhood obesity.Thirty normal-weight and thirty age-and sex-matched obese children were included.Questionnaires and body measurements were collected,and fecal samples underwent 16S rDNA sequencing.Significant differences in body mass index(BMI)and body-fat percentage were observed between the groups.Analysis of gut microbiota diversity revealed lowerα-diversity in obese children.Differences in gut microbiota composition were found between the two groups.Prevotella and Firmicutes were more abundant in the obese group,while Bacteroides and Sanguibacteroides were more prevalent in the control group.AIM To identify the characteristic gut genera in obese and normal-weight children(8-12-year-old)using 16S rDNA sequencing,and provide a basis for subsequent mechanistic studies and prevention strategies for childhood obesity.METHODS Thirty each normal-weight,1:1 matched for age and sex,and obese children,with an obese status from 2020 to 2022,were included in the control and obese groups,respectively.Basic information was collected through questionnaires and body measurements were obtained from both obese and normal-weight children.Fecal samples were collected from both groups and subjected to 16S rDNA sequencing using an Illumina MiSeq sequencing platform for gut microbiota diversity analysis.RESULTS Significant differences in BMI and body-fat percentage were observed between the two groups.The Ace and Chao1 indices were significantly lower in the obese group than those in the control group,whereas differences were not significant in the Shannon and Simpson indices.Kruskal-Wallis tests indicated significant differences in unweighted and weighted UniFrac distances between the gut microbiota of normal-weight and obese children(P<0.01),suggesting substantial disparities in both the species and quantity of gut microbiota between the two groups.Prevotella,Firmicutes,Bacteroides,and Sanguibacteroides were more abundant in the obese and control groups,respectively.Heatmap results demonstrated significant differences in the gut microbiota composition between obese and normal-weight children.CONCLUSION Obese children exhibited lowerα-diversity in their gut microbiota than did the normal-weight children.Significant differences were observed in the composition of gut microbiota between obese and normal-weight children.
文摘Wantong Jingu Tablet(WJT),a mixture of traditional Chinese medicine,was reported to relieve the symptoms of rheumatoid arthritis(RA),but its pharmacological mechanism was not completely understood.The aim of this study was to investigate the therapeutic mechanisms of WJT for RA in vivo.The effects of WJT on joint pathology,as well as the levels of Bax,Bcl-2,caspase-3,cleaved-caspase-3,ERK1/2,pERK1/2,TNF-α,IL-1β,and IL-6 were measured using collagen-induced arthritis(CIA)rats.The intestinal flora composition and the metabolites alteration were analyzed by 16S rDNA sequencing and metabolomics method,respectively.We found that WJT ameliorated the severity of the CIA rats which might be mediated by inducing apoptosis,inactivating the MEK/ERK signals and reducing the production of pro-inflammatory cytokines.WJT,in part,relieved the gut microbiota dysbiosis,especially bacterial phylum Bacteroidetes,Tenericutes and Deferribacteres,as well as bacterial genus Vibrio,Macrococcus and Vagococcus.3’-N-debenzoyl-2’-deoxytaxol,tubulysin B,and magnoline were significantly associated with the specific genera.We identified serotonin,glutathione disulfide,N-acetylneuraminic acid,naphthalene and thromboxane B2 as targeted molecules via metabolomics.Our findings contributed to the understanding of RA pathogenesis,and WJT played essential roles in gut microbiota health and metabolite modulation in the CIA rats.
文摘Dinoflagellates are unicellular eukaryotic protists that dominate in all coastal waters, and are also present in oceanic waters. Despite the central importance of dinoflagellates in global primary production, the relationship between dinoflagellates and bacteria are still poorly understood. In order to understand the ecological interaction between bacterial and dinoflagellates communities, denaturing gradient gel electrophoresis (DGGE) and SSU rRNA sequencing were applied to monitoring the population dynamics of bacteria and dinoflagellates from the onset to disappearance of a dinoflagellates bloom occurred in Baltimore Inner Harbor, from April 15 to 24,2002. Although Prorocentrum minimum was the major bloom forming species under the light microscopy, DGGE method with dinoflagellate specific primers demonstrated that Prorocentrum micans, Gymnodinium galatheanum and Gyrodinium uncatenum were also present during the bloom. Population shifts among the minor dinoflagellate groups were observed. DGGE of PCR-amplified 16S rRNA gene fragments indicated that cyanobacteria, α, β, γ-proteobacteria, Flavobacterium Bacteroides-Cytophaga (FBC), and Planctomcetes were the major components of bacterial assemblages during the bloom. DGGE analysis showed that Cytophagales and α-proteobacteria played important roles at different stages of dinoflagellates bloom. DGGE can be used as a rapid tool to simultaneously monitor population dynamics of both bacterial and dinoflagellates communities in aquatic environments, which is demonstrated here.
基金This research was supported by Basic Science Research Program of the National Research Foundation of Korea(NRF)in the Ministry of ScienceICT and Future Planning(NRF-2017R1A2B4005915)supported this research.
文摘We investigated the diversity and composition of microflora in feces of Lycopus lucidus Turcz.-fed mice.In addition,we evaluated the production of major cytokines(Interleukin-6 and-10)which are related to inflammation and fatty acid composition of several tissues.16S ribosomal DNA sequencing-based microbiome taxonomic profiling analysis was performed utilizing the EzBioCloud data base.Male mice fed on L.lucidus showed a significantly reduced number of lactic acid bacteria and coliform in the feces compared with the control group(p<0.05).16S rDNA sequencing analysis of fecal samples showed that L.lucidus supplementation decreased the community of harmful microflora(Enterobacteriaceae including Escherichia coli and Bacteroides sp.)in feces compared with the control group(p<0.05).There were no significant differences in mRNA expression of cytokine IL-6 and IL-10 between the control and L.lucidus fed groups.The fecal fatty acid composition in the L.lucidus group had percentages of 4:0,6:0,8:0 and 10:0 in the intestine but those short chain fatty acids were not detected in the control group.Our results showed that L.lucidus supplementation influenced gut environment by decreasing harmful microflora and increased the percentages of several short fatty acids.
基金supported by the National Natural Science Foundation of China(81373769)the Scientific Research and Postgraduate Training Co-construction Project of Beijing Municipal Education Commission(1000062520115)the Province Natural Science Foundation of Beijing(7172131)。
文摘Objective:To explore the structural and functional characteristics of microbiota in oropharynx of subhealthy children with gastrointestinal heat retention syndrome(GHRS)differentiated by traditional Chinese medicine,and screen the biometric operational taxonomic units(OTUs)to assist the clinical diagnosis.Methods:We recruited children according to the“GHRS diagnostic scale”,collected their oropharyngeal swabs,and sequenced the 16 SrDNA V4 region.We described the bacterial structure with alpha-indexes,beta-distances,and relative abundances;moreover,we screened the differential genera/OTUs with Wilcoxon rank-sum test,Metagenome Seq analysis,and linear discriminant analysis effect size(LEf Se)analysis,in which biometric OTUs were selected to construct the receiver operating characteristic curve to verify the diagnostic value.The bacterial function was predicted with Kyoto Encyclopedia of Genes and Genomes pathways according to 16S rDNA gene by using Phylogenetic Investigation of Communities by Reconstruction of Unobserved States.Results:The study population was composed of 10 children with GHRS and 10 healthy control children.GHRS children were more likely to overeat(gluttony,P=.033).Alpha-indexes,such as Sobs,abundancebased coverage estimator,Bootstrap,and Qstat,were significantly higher in the GHRS group,while betadistances did not exhibit any significant intergroup differences.There were 9 differently distributed nonpredominant genera between the groups in Wilcoxon rank-sum test,as well as 13 non-predominant genera in Metagenome Seq analysis and 3 non-predominant OTUs in LEfSe analysis.OTU44 and OTU196 were used to construct the receiver operating characteristic curve,and the area under curve was 0.92.Predicted functions showed that pathways related to oxidative phosphorylation and carbon metabolism were enriched in healthy control samples,while the pathway related to renin secretion was remarkably enriched in GHRS samples.Conclusion:Unique oropharyngeal microbial structure and function were identified in GHRS children.OTU44 and OTU196 were specific OTUs,which could be used as biomarkers of GHRS to assist clinical diagnosis.
