Analysis of the Rumen Bacterial Diversity under two Different Diet Conditions using Denaturing Gradient Gel Electrophoresis, Random Sequencing, and Statistical Ecology Approaches

Anaerobe ◽  
2001 ◽  
Vol 7 (3) ◽  
pp. 119-134 ◽  
Author(s):  
Svetlana A. Kocherginskaya ◽  
Rustam I. Aminov ◽  
Bryan A. White
Keyword(s):  
Bacterial Diversity ◽  
Gel Electrophoresis ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Gradient Gel Electrophoresis ◽  
Random Sequencing ◽  
Statistical Ecology

scholarly journals 16S Ribosomal DNA-Based Analysis of Bacterial Diversity in Purified Water Used in Pharmaceutical Manufacturing Processes by PCR and Denaturing Gradient Gel Electrophoresis

2002 ◽  
Vol 68 (2) ◽  
pp. 699-704 ◽  
Author(s):  
Mako Kawai ◽  
Eiichi Matsutera ◽  
Hisashi Kanda ◽  
Nobuyasu Yamaguchi ◽  
Katsuji Tani ◽  
...  
Keyword(s):  
Bacterial Diversity ◽  
Ribosomal Dna ◽  
Gel Electrophoresis ◽  
Esterase Activity ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Pharmaceutical Manufacturing ◽  
Manufacturing Processes ◽  
Culturable Bacteria ◽  
16S Ribosomal Dna ◽  
Gradient Gel Electrophoresis

ABSTRACT The bacterial community in partially purified water, which is prepared by ion exchange from tap water and is used in pharmaceutical manufacturing processes, was analyzed by denaturing gradient gel electrophoresis (DGGE). 16S ribosomal DNA fragments, including V6, -7, and -8 regions, were amplified with universal primers and analyzed by DGGE. The bacterial diversity in purified water determined by PCR-DGGE banding patterns was significantly lower than that of other aquatic environments. The bacterial populations with esterase activity sorted by flow cytometry and isolated on soybean casein digest (SCD) and R2A media were also analyzed by DGGE. The dominant bacterium in purified water possessed esterase activity but could not be detected on SCD or R2A media. DNA sequence analysis of the main bands on the DGGE gel revealed that culturable bacteria on these media were Bradyrhizobium sp., Xanthomonas sp., and Stenotrophomonas sp., while the dominant bacterium was not closely related to previously characterized bacteria. These data suggest the importance of culture-independent methods of quality control for pharmaceutical water.

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