This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Kawai, M.
Right arrow Articles by Nasu, M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Kawai, M.
Right arrow Articles by Nasu, M.
Agricola
Right arrow Articles by Kawai, M.
Right arrow Articles by Nasu, M.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, February 2002, p. 699-704, Vol. 68, No. 2
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.2.699-704.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

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

Mako Kawai,1 Eiichi Matsutera,1 Hisashi Kanda,1 Nobuyasu Yamaguchi,2 Katsuji Tani,2 and Masao Nasu2*

Quality Assurance Division, Biological Quality Assurance Section, Dainippon Pharmaceutical Co. Ltd., Osaka 553-0001 and ,1 Graduate School of Pharmaceutical Science, Osaka University, Osaka 565-0871, Japan2

Received 21 September 2001/ Accepted 26 November 2001

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.

* Corresponding author. Mailing address: Graduate School of Pharmaceutical Science, Osaka University, 1-6, Yamada-oka, Suita, Osaka 565-0871, Japan. Phone 81-6879-8170. Fax: 81-6879-8174. E-mail: nasu{at}phs.osaka-u.ac.jp.

Applied and Environmental Microbiology, February 2002, p. 699-704, Vol. 68, No. 2
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.2.699-704.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Yamaguchi, N., Baba, T., Nakagawa, S., Saito, A., Nasu, M. (2007). Rapid monitoring of bacteria in dialysis fluids by fluorescent vital staining and microcolony methods. Nephrol Dial Transplant 22: 612-616 [Full Text]  
  • Alexeeva, I., Elliott, E. J., Rollins, S., Gasparich, G. E., Lazar, J., Rohwer, R. G. (2006). Absence of Spiroplasma or Other Bacterial 16S rRNA Genes in Brain Tissue of Hamsters with Scrapie. J. Clin. Microbiol. 44: 91-97 [Abstract] [Full Text]  
  • Hoefel, D., Monis, P. T., Grooby, W. L., Andrews, S., Saint, C. P. (2005). Culture-Independent Techniques for Rapid Detection of Bacteria Associated with Loss of Chloramine Residual in a Drinking Water System. Appl. Environ. Microbiol. 71: 6479-6488 [Abstract] [Full Text]  
  • Yu, Z., Morrison, M. (2004). Comparisons of Different Hypervariable Regions of rrs Genes for Use in Fingerprinting of Microbial Communities by PCR-Denaturing Gradient Gel Electrophoresis. Appl. Environ. Microbiol. 70: 4800-4806 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»