کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
10889913 | 1081551 | 2005 | 8 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Improved permeabilization protocols for fluorescence in situ hybridization (FISH) of mycolic-acid-containing bacteria found in foams
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کلمات کلیدی
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
بیوتکنولوژی یا زیستفناوری
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![عکس صفحه اول مقاله: Improved permeabilization protocols for fluorescence in situ hybridization (FISH) of mycolic-acid-containing bacteria found in foams Improved permeabilization protocols for fluorescence in situ hybridization (FISH) of mycolic-acid-containing bacteria found in foams](/preview/png/10889913.png)
چکیده انگلیسی
Formation of thick, stable foams and scums on activated sludge wastewater treatment plants is a worldwide problem, and to better understand what causes this foam and to cure it, there is a need to identify and quantify the bacteria present there. Fluorescence in situ hybridisation (FISH) overcomes the difficulties experienced with microscopic methods of identification for the mycolic-acid-containing actinomycetes (the mycolata), which are present in foams, where many share the morphotype of right-angled branching filaments. However, the presence of hydrophobic mycolic acids in their cell wall makes this group of bacteria particularly difficult to permeabilise, which greatly reduces the usefulness of FISH. While several permeabilisation treatments have been described, none appear to adequately permeabilise all genera of the mycolata. In this study several protocols for permeabilisation were assessed with both pure cultures of selected genera of the mycolata and foam samples. Combining mild acid hydrolysis with enzyme treatments (either mutanolysin/lysozyme or lipase/proteinase K) was found to be the most effective method, although other evidence presented here suggests that negative FISH results can not always be explained in terms of cell permeability to the probes.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Microbiological Methods - Volume 61, Issue 1, April 2005, Pages 47-54
Journal: Journal of Microbiological Methods - Volume 61, Issue 1, April 2005, Pages 47-54
نویسندگان
Emma L. Carr, Kathryn Eales, Jacques Soddell, Robert J. Seviour,