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J Microbiol Methods ; 88(2): 297-303, 2012 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-22230233

RESUMO

Surface associated microbes have historically been difficult to accurately and effectively enumerate. In the current study, we propose a rapid and simple method for estimating abundance of surface associated microbial cells by fluorescence of SYBRGreen stained bacteria and in vivo chlorophyll a fluorescence of benthic diatoms in 24 and 48-well microtiter plates. The effectiveness of this high-throughput technique is demonstrated by assessing sensitivity of a clinical strain of Vibrio cholerae, a benthic bacterial isolate and the benthic microalgae Cylindrotheca closterium to three antibiotics--tylosin, lincomycin and ciproflaxacin. We report on the significant linear relationships between spectral chl a fluorescence and cell abundance and between microalgal growth rates derived from cell counts and fluorescence. Additionally, we provide a simplified and improved method for preparation of a silica gel matrix (SGM), which is an ideal plating media for fluorescence applications. These findings indicate that spectrofluorometry is an inexpensive tool for rapidly estimating abundance of surface associated microbiota and can be employed for assessing antibiotic sensitivity.


Assuntos
Carga Bacteriana/métodos , Diatomáceas/microbiologia , Microalgas/microbiologia , Espectrometria de Fluorescência/métodos , Análise de Variância , Benzotiazóis , Clorofila/análise , Clorofila/química , Clorofila A , Meios de Cultura , Diaminas , Corantes Fluorescentes/química , Ensaios de Triagem em Larga Escala/métodos , Testes de Sensibilidade Microbiana , Compostos Orgânicos/química , Quinolinas , Reprodutibilidade dos Testes , Sílica Gel/química , Vibrio cholerae/citologia
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