Comparison of methods for the concentration of bacterioplankton for in situ hybridization

• Published in: Journal of microbiological methods Vol. 29; no. 1; pp. 23 - 29
• Main Authors: Lemke, Michael J, McNamara, Christopher J, G. Leff, Laura
• Format: Journal Article
• Language: English
• Published: Shannon Elsevier B.V 01.04.1997; Elsevier Science
• Subjects: Bacteriological methods and techniques used in bacteriology; Bacteriology; Bacterioplankton; Biological and medical sciences; Burkholderia cepacia; Concentration; Fundamental and applied biological sciences. Psychology; Hybridization; In situ; Microbiology; Bacterioplankton; Concentration; Hybridization; Burkholderia cepacia; In situ; Freshwater environment; Molecular hybridization; Membrane filtration; Bacteria; Method; Comparative study
• ISSN: 0167-7012; 1872-8359
• DOI: 10.1016/S0167-7012(97)00989-5

Three approaches for concentrating stream bacterioplankton were compared for analysis by in situ hybridization: centrifugation; filtration onto a polymer membrane and transfer to a gel-coated slide; and filtration onto an inorganic filter disk. Stream bacterioplankton were hybridized with a species-specific oligonucleotide probe targeted for 23S rRNA of Burkholderia cepacia for analysis of variability among techniques. The inorganic filter technique yielded three to six times higher total cell counts and appeared to be effected less by interfering suspended particles and differences in site conditions than the other two concentration methods. Hybridization of samples with minimum cell loss yielded different, and presumably more accurate, estimates of the relative abundance of B. cepacia than the other methods.