Information on co-adherence of different oral bacterial species is important for understanding interspecies interactions within oral microbial community. Current knowledge on this topic is heavily based on pariwise co...Information on co-adherence of different oral bacterial species is important for understanding interspecies interactions within oral microbial community. Current knowledge on this topic is heavily based on pariwise coaggregation of known, cultivable species. In this study, we employed a membrane binding assay coupled with polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) to systematically analyze the co-adherence profiles of oral bacterial species, and achieved a more profound knowledge beyond pairwise coaggregation. Two oral bacterial species were selected to serve as "bait": Fusobacterium nucleatum (F. nucleatum) whose ability to adhere to a multitude of oral bacterial species has been extensively studied for pairwise interactions and Streptococcus mutans (S. mutans) whose interacting partners are largely unknown. To enable screening of interacting partner species within bacterial mixtures, cells of the "bait" oral bacterium were immobilized on nitrocellulose membranes which were washed and blocked to prevent unspecific binding. The "prey" bacterial mixtures (including known species or natural saliva samples) were added, unbound cells were washed off after the incubation period and the remaining cells were eluted using 0.2 mol·L-1 glycine. Genomic DNA was extracted, subjected to 16S rRNA PCR amplification and separation of the resulting PCR products by DGGE. Selected bands were recovered from the gel, sequenced and identified via Nucleotide BLAST searches against different databases. While few bacterial species bound to S. mutans, consistent with previous findings F. nucleatum adhered to a variety of bacterial species including uncultivable and uncharacterized ones. This new approach can more effectively analyze the co-adherence profiles of oral bacteria, and could facilitate the systematic study of interbacterial binding of oral microbial species.展开更多
The actinomycete populations and functions in cadmium (Cd) contaminated soil were investigated by the cultivationindependent molecular methods.The genomic DNA was extracted and purified from soil adulterated with vari...The actinomycete populations and functions in cadmium (Cd) contaminated soil were investigated by the cultivationindependent molecular methods.The genomic DNA was extracted and purified from soil adulterated with various concentrations of Cd in the laboratory.The partial 16S rDNA genes were amplified by polymerase chain reaction (PCR) using specific primers bound to evolutionarily conserved regions within these actinomycete genes.The diversity in PCRamplified products,as measured by denaturing gradient gel electrophoresis (EGGE),was used as a genetic fingerprint of the population.Principle component analysis and Shannon-Weaver diversity index (H) analyses were used to analyze the DGGE results.Results showed that the two principal components accounted for only a low level of the total variance.The value H in contaminated soil was lower than that in the control at later stages of cultivation,whereas at earlier stages it was higher.Among the six sampling time points,the first,fifth and sixth weeks had the highest values of H.Significantly negative correlations between bioavailable Cd concentration and H values existed in the samples from weeks 2 (R=0.929,P < 0.05) and 4 (R=0.909,P < 0.05).These results may shed light on the effect of Cd on the soil environment and the chemical behavior and toxicity of Cd to actinomycetes.展开更多
To study the structure of microbial communities in the biological hydrogen produc- tion reactor and determine the ecological function of hydrogen producing bacteria, anaerobic sludge was obtained from the continuous s...To study the structure of microbial communities in the biological hydrogen produc- tion reactor and determine the ecological function of hydrogen producing bacteria, anaerobic sludge was obtained from the continuous stirred tank reactor (CSTR) in different periods of time, and the diversity and dynamics of microbial communities were investigated by denaturing gra- dient gel electrophoresis (DGGE). The results of DGGE demonstrated that an obvious shift of microbial population happened from the beginning of star-up to the 28th day, and the ethanol type fermentation was established. After 28 days the structure of microbial community became stable, and the climax community was formed. Comparative analysis of 16S rDNA sequences from reamplifying and sequencing the prominent bands indicated that the dominant population belonged to low G+C Gram-positive bacteria (Clostridium sp. and Ethanologenbacterium sp.), β- proteobacteria (Acidovorax sp.), γ-proteobacteria (Kluyvera sp.), Bacteroides (uncultured bacte- rium SJA-168), and Spirochaetes (uncultured eubacterium E1-K13), respectively. The hydrogen production rate increased obviously with the increase of Ethanologenbacterium sp., Clostridium sp. and uncultured Spirochaetes after 21 days, meanwhile the succession of ethanol type fer- mentation was formed. Throughout the succession the microbial diversity increased however it decreased after 21 days. Some types of Clostridium sp. Acidovorax sp., Kluyvera sp., and Bac- teroides were dominant populations during all periods of time. These special populations were essential for the construction of climax community. Hydrogen production efficiency was de- pendent on both hydrogen producing bacteria and other populations. It implied that the co- metabolism of microbial community played a great role of biohydrogen production in the reactors.展开更多
Bacterial diversity was investigated in soil samples collected from 13 sites around the Great Wall Station, Fildes Peninsula, King George Island, Antarctica, using denaturing gradient gel electrophoresis (DGGE) of 16S...Bacterial diversity was investigated in soil samples collected from 13 sites around the Great Wall Station, Fildes Peninsula, King George Island, Antarctica, using denaturing gradient gel electrophoresis (DGGE) of 16S rRNA genes. The classes α-, β-, and γ-Proteobacteria, as well as the phylum Actinobacteria, were found to be the dominant bacteria in the soils around the Great Wall Station. Although the selected samples were not contaminated by oil, a relationship between soil parameters, microbial biodiversity, and human impact was still seen. Sample sites in human impacted areas showed lower bacterial biodiversity (average H′= 2.65) when compared to nonimpacted sites (average H′= 3.05). There was no statistically significant correlation between soil bacterial diversity and total organic carbon (TOC), total nitrogen, or total phosphorus contents of the soil. Canonical correlation analysis showed that TOC content was the most important factor determining bacterial community profiles among the measured soil parameters. In conclusion, microbial biodiversity and community characteristics within relatively small scales (1.5 km) were determined as a function of local environment parameters and anthropogenic impact.展开更多
基金This work was supported by the National Basic Research Program （973） of China （No. 2002CB412307） the Hi-Tech Research and Development Program （863） of China （No. 2002AA601011） the National Natural Science Foundation of China （No. 40371102）.
基金supported by Chinese State Scholarship Fund to R. WangUS National Institutes of Health (NIH) Grants DE020102 and GM95373 to W. Shi
文摘Information on co-adherence of different oral bacterial species is important for understanding interspecies interactions within oral microbial community. Current knowledge on this topic is heavily based on pariwise coaggregation of known, cultivable species. In this study, we employed a membrane binding assay coupled with polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) to systematically analyze the co-adherence profiles of oral bacterial species, and achieved a more profound knowledge beyond pairwise coaggregation. Two oral bacterial species were selected to serve as "bait": Fusobacterium nucleatum (F. nucleatum) whose ability to adhere to a multitude of oral bacterial species has been extensively studied for pairwise interactions and Streptococcus mutans (S. mutans) whose interacting partners are largely unknown. To enable screening of interacting partner species within bacterial mixtures, cells of the "bait" oral bacterium were immobilized on nitrocellulose membranes which were washed and blocked to prevent unspecific binding. The "prey" bacterial mixtures (including known species or natural saliva samples) were added, unbound cells were washed off after the incubation period and the remaining cells were eluted using 0.2 mol·L-1 glycine. Genomic DNA was extracted, subjected to 16S rRNA PCR amplification and separation of the resulting PCR products by DGGE. Selected bands were recovered from the gel, sequenced and identified via Nucleotide BLAST searches against different databases. While few bacterial species bound to S. mutans, consistent with previous findings F. nucleatum adhered to a variety of bacterial species including uncultivable and uncharacterized ones. This new approach can more effectively analyze the co-adherence profiles of oral bacteria, and could facilitate the systematic study of interbacterial binding of oral microbial species.
基金Project supported by the National Natural Science Foundation of China (Nos. 30570053 and 40501037)the National High Technology Research and Development Program (863 Program) of China (No. 2007AA10Z409)+1 种基金the National"Eleventh Five Years Plan" Key Project on Science and Technology of China (No. 2006BAJ08B01)the Research Fund of Science and Technology Bureau of Zhejiang Province,China (No. 2008C23088)
文摘The actinomycete populations and functions in cadmium (Cd) contaminated soil were investigated by the cultivationindependent molecular methods.The genomic DNA was extracted and purified from soil adulterated with various concentrations of Cd in the laboratory.The partial 16S rDNA genes were amplified by polymerase chain reaction (PCR) using specific primers bound to evolutionarily conserved regions within these actinomycete genes.The diversity in PCRamplified products,as measured by denaturing gradient gel electrophoresis (EGGE),was used as a genetic fingerprint of the population.Principle component analysis and Shannon-Weaver diversity index (H) analyses were used to analyze the DGGE results.Results showed that the two principal components accounted for only a low level of the total variance.The value H in contaminated soil was lower than that in the control at later stages of cultivation,whereas at earlier stages it was higher.Among the six sampling time points,the first,fifth and sixth weeks had the highest values of H.Significantly negative correlations between bioavailable Cd concentration and H values existed in the samples from weeks 2 (R=0.929,P < 0.05) and 4 (R=0.909,P < 0.05).These results may shed light on the effect of Cd on the soil environment and the chemical behavior and toxicity of Cd to actinomycetes.
基金This work was supported by the National Science Foundation for Distinguished Young Scholars(No.50125823)National Natural Science Foundation of China(Grant No.30470054)Key Project of Chinese National Programs for Fundamental Research and Development(No.G2000026402).
文摘To study the structure of microbial communities in the biological hydrogen produc- tion reactor and determine the ecological function of hydrogen producing bacteria, anaerobic sludge was obtained from the continuous stirred tank reactor (CSTR) in different periods of time, and the diversity and dynamics of microbial communities were investigated by denaturing gra- dient gel electrophoresis (DGGE). The results of DGGE demonstrated that an obvious shift of microbial population happened from the beginning of star-up to the 28th day, and the ethanol type fermentation was established. After 28 days the structure of microbial community became stable, and the climax community was formed. Comparative analysis of 16S rDNA sequences from reamplifying and sequencing the prominent bands indicated that the dominant population belonged to low G+C Gram-positive bacteria (Clostridium sp. and Ethanologenbacterium sp.), β- proteobacteria (Acidovorax sp.), γ-proteobacteria (Kluyvera sp.), Bacteroides (uncultured bacte- rium SJA-168), and Spirochaetes (uncultured eubacterium E1-K13), respectively. The hydrogen production rate increased obviously with the increase of Ethanologenbacterium sp., Clostridium sp. and uncultured Spirochaetes after 21 days, meanwhile the succession of ethanol type fer- mentation was formed. Throughout the succession the microbial diversity increased however it decreased after 21 days. Some types of Clostridium sp. Acidovorax sp., Kluyvera sp., and Bac- teroides were dominant populations during all periods of time. These special populations were essential for the construction of climax community. Hydrogen production efficiency was de- pendent on both hydrogen producing bacteria and other populations. It implied that the co- metabolism of microbial community played a great role of biohydrogen production in the reactors.
基金supported by the National Natural Science Foundation of China (No. 51108331)the Chinese Polar Environment Comprehensive Investigation and Assessment Programmes (No. CHINARE2012-02-01-08, CHINARE2013-02-01-08, CHINARE2013-04-01-07)
文摘Bacterial diversity was investigated in soil samples collected from 13 sites around the Great Wall Station, Fildes Peninsula, King George Island, Antarctica, using denaturing gradient gel electrophoresis (DGGE) of 16S rRNA genes. The classes α-, β-, and γ-Proteobacteria, as well as the phylum Actinobacteria, were found to be the dominant bacteria in the soils around the Great Wall Station. Although the selected samples were not contaminated by oil, a relationship between soil parameters, microbial biodiversity, and human impact was still seen. Sample sites in human impacted areas showed lower bacterial biodiversity (average H′= 2.65) when compared to nonimpacted sites (average H′= 3.05). There was no statistically significant correlation between soil bacterial diversity and total organic carbon (TOC), total nitrogen, or total phosphorus contents of the soil. Canonical correlation analysis showed that TOC content was the most important factor determining bacterial community profiles among the measured soil parameters. In conclusion, microbial biodiversity and community characteristics within relatively small scales (1.5 km) were determined as a function of local environment parameters and anthropogenic impact.
