Detection and quantification of Vibrio populations using denaturant gradient gel electrophoresis

2006 ◽  
Vol 67 (2) ◽  
pp. 339-348 ◽  
Author(s):  
Alexander Eiler ◽  
Stefan Bertilsson
2012 ◽  
Vol 52 (7) ◽  
pp. 647 ◽  
Author(s):  
E. D. Martínez ◽  
K. E. Turnbull ◽  
S. P. Quigley ◽  
S. J. Streeter ◽  
A. Swain ◽  
...  

Two experiments were conducted to evaluate the relationship between rumen liquid-associated bacterial community structures and post-weaning liveweight gain (LWG) of Brahman crossbred steers. Bacterial diversity was assessed using denaturant gradient gel electrophoresis (DGGE). In Experiment 1, 16 steers were selected from a group of 100 steers by pairing steers with the same weaning weight, but different LWG 90 days after weaning (n = 8 highest growth, 0.21 ± 0.01 kg/day; and n = 8 lowest growth –0.07 ± 0.01 kg/day). Thereafter, steers were allocated to a 28-day pen study and fed Mitchell grass (Astrebla spp.) hay to examine DM intake and digestibility, rumen parameters and rumen microbial community in these two groups. Rumen fluid samples were taken by stomach tube 3 h after feeding on the last day of the pen phase. In Experiment 2, 12 pairs of weaned steers were selected from a group of 203 steers on the same basis as Experiment 1. The post-weaning LWG were 0.20 ± 0.03 and 0.02 ± 0.03 kg/day for the 12 highest and 12 lowest growth animals selected, respectively. Steers then grazed dry season Sabi grass (Urochloa mosambicensis) dominant pasture for 21 days, before rumen sampling 3 h after morning grazing by stomach tubing on the last day. In Experiment 1, there were no significant differences between the two groups in DM intake, digestibility, ruminal pH, total volatile fatty acid (VFA) concentration or the VFA proportion. In Experiments 1 and 2, rumen ammonia-N concentration was similar between LWG groups and there was no evidence of a relationship between liquid-phase DGGE profiles of rumen bacteria and high or low post-weaning LWG using multivariate analyses. Furthermore, the number of detected DGGE bands, the Shannon–Wiener and evenness indexes were not different between LWG groups. This DGGE analysis of the most abundant groups of rumen fluid-associated bacteria suggests that microbial populations were not related with the differences observed in post-weaning LWG within a group of weaners fed low crude protein diets.


2004 ◽  
Vol 385 (10) ◽  
pp. 967-973 ◽  
Author(s):  
Masahiko Shiraishi ◽  
Adam J. Oates ◽  
Xu Li ◽  
Ying H. Chuu ◽  
Takao Sekiya

Abstract The technique of segregation of partly melted molecules (SPM) is a convenient and efficient method to isolate DNA fragments associated with CpG islands. The approach is conceptually simple and uses denaturant gradient gel electrophoresis to separate DNA molecules digested with restriction endonucleases. The SPM methodology has successfully been applied to the identification of genes from anonymous, unsequenced DNA fragments and CpG islands methylated in human cancer. In this article the theoretical background and practical application of the SPM method is reviewed.


2004 ◽  
Vol 70 (6) ◽  
pp. 3789-3793 ◽  
Author(s):  
Thomas E. Freitag ◽  
James I. Prosser

ABSTRACT To assess links between betaproteobacterial ammonia-oxidizing bacteria (AOB) in marine sediment and in overlying water, communities in Loch Duich, Scotland, were characterized by analysis of clone libraries and denaturant gradient gel electrophoresis of 16S rRNA gene fragments. Nitrosospira cluster 1-like sequences were isolated from both environments, but different sequence types dominated water and sediment samples. Detailed phylogenetic analysis of marine Nitrosospira cluster 1-like sequences in Loch Duich and surrounding regions suggests the existence of at least two different phylogenetic subgroups, potentially indicative of new lineages within the betaproteobacterial AOB, representing different marine ecotypes.


2020 ◽  
Vol 51 (2) ◽  
pp. 125-146
Author(s):  
Nasiruddin Nasiruddin ◽  
Yu Zhangxin ◽  
Ting Zhao Chen Guangying ◽  
Minghui Ji

We grew cucumber in pots in greenhouse for 9-successive cropping cycles and analyzed the rhizosphere Pseudomonas spp. community structure and abundance by PCR-denaturing gradient gel electrophoresis and quantitative PCR. Results showed that continuous monocropping changed the cucumber rhizosphere Pseudomonas spp. community. The number of DGGE bands, Shannon-Wiener index and Evenness index decreased during the 3rd cropping and thereafter, increased up to the 7th cropping, however, however, afterwards they decreased again. The abundance of Pseudomonas spp. increased up to the 5th successive cropping and then decreased gradually. These findings indicated that the structure and abundance of Pseudomonas spp. community changed with long-term cucumber monocropping, which might be linked to soil sickness caused by its continuous monocropping.


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