scholarly journals Comparison of Fecal Methanogenic Archaeal Community Between Erhualian and Landrace Pigs Using Denaturing Gradient Gel Electrophoresis and Real-Time PCR Analysis

2014 ◽  
Vol 13 (6) ◽  
pp. 1340-1348 ◽  
Author(s):  
Yong SU ◽  
Hauke Smidt ◽  
Wei-Yun ZHU
2007 ◽  
Vol 73 (18) ◽  
pp. 5731-5741 ◽  
Author(s):  
Beatrice Vitali ◽  
Ciro Pugliese ◽  
Elena Biagi ◽  
Marco Candela ◽  
Silvia Turroni ◽  
...  

ABSTRACT The microbial flora of the vagina plays a major role in preventing genital infections, including bacterial vaginosis (BV) and candidiasis (CA). An integrated approach based on PCR-denaturing gradient gel electrophoresis (PCR-DGGE) and real-time PCR was used to study the structure and dynamics of bacterial communities in vaginal fluids of healthy women and patients developing BV and CA. Universal eubacterial primers and Lactobacillus genus-specific primers, both targeted at 16S rRNA genes, were used in DGGE and real-time PCR analysis, respectively. The DGGE profiles revealed that the vaginal flora was dominated by Lactobacillus species under healthy conditions, whereas several potentially pathogenic bacteria were present in the flora of women with BV. Lactobacilli were the predominant bacterial population in the vagina for patients affected by CA, but changes in the composition of Lactobacillus species were observed. Real-time PCR analysis allowed the quantitative estimation of variations in lactobacilli associated with BV and CA diseases. A statistically significant decrease in the relative abundance of lactobacilli was found in vaginal fluids of patients with BV compared to the relative abundance of lactobacilli in the vaginal fluids of healthy women and patients with CA.


2016 ◽  
Vol 46 (2) ◽  
pp. 190-199 ◽  
Author(s):  
Marliane de Cássia Soares da Silva ◽  
Igor Rodrigues Mendes ◽  
Thiago de Almeida Paula ◽  
Roberto Sousa Dias ◽  
Sérgio Oliveira de Paula ◽  
...  

A large proportion of eucalypt plantations in Brazil are located in areas with low soil fertility. The actions of microorganisms are of great importance for the cycling of nutrients, including nitrogen (N), that are essential for plant metabolism. Denaturing gradient gel electrophoresis (DGGE) was used to monitor and identify the total and active microorganisms involved in the N cycle in both the soil and root systems of a forest of Eucalyptus urograndis with sections that were fertilized with N or unfertilized. Quantitative real-time PCR was used to examine the expression of the nifH gene in N-fixing bacteria present in both the soil and root systems. According to the DGGE analysis, in the total and active populations of N-fixing bacteria, the presence and expression of the nifH gene were influenced by the winter and summer seasons and (or) N fertilization, respectively. DGGE band sequencing from total DNA samples showed that the most abundant group of diazotrophic bacteria belonged to Alphaproteobacteria in both the soil and root systems. Quantitative real-time PCR revealed that nifH expression was higher in the soil samples, especially in those that did not receive N fertilization. The differences in the composition of the total and active diazotrophic populations highlight the importance of evaluating the active populations, because they are effectively responsible for the biogeochemical transformation of N and also control its’ availability to plants.


2011 ◽  
Vol 57 (6) ◽  
pp. 504-513 ◽  
Author(s):  
Min Liu ◽  
Tian Xiao ◽  
Ying Wu ◽  
Feng Zhou ◽  
Wuchang Zhang

The archaeal community and the effects of environmental factors on microbial community distribution were investigated at five sampling sites in the Changjiang Estuary hypoxia area and the adjacent East China Sea in June, August, and October 2006. Profiles of the archaeal communities were generated by denaturing gradient gel electrophoresis of 16S rRNA genes followed by DNA sequence analysis, and the results were analyzed by multivariate statistical analysis. Denaturing gradient gel electrophoresis band patterns were analyzed by cluster analysis to assess temporal changes in the genetic diversity of the archaeal communities. Most of the October samples grouped together separately from those of June and August. Analysis of DNA sequences revealed that the dominant archaeal groups in the Changjiang Estuary hypoxia area and the adjacent East China Sea were affiliated with Euryarchaeota (mainly marine group II) and Crenarchaeota. The effects of environmental factors on the archaeal community distribution were analyzed by the ordination technique of canonical correspondence analysis. Salinity had a significant effect on the archaeal community composition.


2000 ◽  
Vol 66 (8) ◽  
pp. 3269-3276 ◽  
Author(s):  
Eddie Cytryn ◽  
Dror Minz ◽  
Ronald S. Oremland ◽  
Yehuda Cohen

ABSTRACT The vertical and seasonal distribution and diversity of archaeal sequences was investigated in a hypersaline, stratified, monomictic lake, Solar Lake, Sinai, Egypt, during the limnological development of stratification and mixing. Archaeal sequences were studied via phylogenetic analysis of 16S rDNA sequences as well as denaturing gradient gel electrophoresis analysis. The 165 clones studied were grouped into four phylogenetically different clusters. Most of the clones isolated from both the aerobic epilimnion and the sulfide-rich hypolimnion were defined as cluster I, belonging to theHalobacteriaceae family. The three additional clusters were all isolated from the anaerobic hypolimnion. Cluster II is phylogenetically located between the generaMethanobacterium and Methanococcus. Clusters III and IV relate to two previously documented groups of uncultured euryarchaeota, remotely related to the genusThermoplasma. No crenarchaeota were found in the water column of the Solar Lake. The archaeal community in the Solar Lake under both stratified and mixed conditions was dominated by halobacteria in salinities higher than 10%. During stratification, additional clusters, some of which may possibly relate to uncultured halophilic methanogens, were found in the sulfide- and methane-rich hypolimnion.


2003 ◽  
Vol 69 (1) ◽  
pp. 475-482 ◽  
Author(s):  
Christiane B. Meroth ◽  
Jens Walter ◽  
Christian Hertel ◽  
Markus J. Brandt ◽  
Walter P. Hammes

ABSTRACT Four sourdoughs (A to D) were produced under practical conditions by using a starter mixture of three commercially available sourdough starters and a baker's yeast constitutively containing various species of lactic acid bacteria (LAB). The sourdoughs were continuously propagated until the composition of the LAB flora remained stable. Two LAB-specific PCR-denaturing gradient gel electrophoresis (DGGE) systems were established and used to monitor the development of the microflora. Depending on the prevailing ecological conditions in the different sourdough fermentations, only a few Lactobacillus species were found to be competitive and became dominant. In sourdough A (traditional process with rye flour), Lactobacillus sanfranciscensis and a new species, L. mindensis, were detected. In rye flour sourdoughs B and C, which differed in the process temperature, exclusively L. crispatus and L. pontis became the predominant species in sourdough B and L. crispatus, L. panis, and L. frumenti became the predominant species in sourdough C. On the other hand, in sourdough D (corresponding to sourdough C but produced with rye bran), L. johnsonii and L. reuteri were found. The results of PCR-DGGE were consistent with those obtained by culturing, except for sourdough B, in which L. fermentum was also detected. Isolates of the species L. sanfranciscensis and L. fermentum were shown by randomly amplified polymorphic DNA-PCR analysis to originate from the commercial starters and the baker's yeast, respectively.


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