Scheda di dettaglio – i prodotti della ricerca
| Dato | Valore |
|---|---|
| Title | BACTERIAL GROWTH KINETICS ESTIMATION BY FLUORESCENCE IN SITU HYBRIDIZATION AND SPECTROFLUOROMETRIC QUANTIFICATION |
| Abstract | Aims: The aim of this study was to develop a specific and rapid method to identify and quantify relevant bacterial populations in mixed biomass by spectrofluorometric quantification (SQ) of whole cells hybridized with fluorescently labelled oligonucleotide probes targeting mature 16S ribosomal RNA (rRNA). Probe targeting the precursor of rRNA synthesis was also employed because it was being suggested as more indicative of the activity state of the microorganisms. Methods and Results: Original fluorescence in situ hybridization protocol was modified to be applied to liquid samples and the fluorescence emission from the Cy3-labelled cells was measured by spectrofluorometry. The method was calibrated on an exponentially growing cell suspension of Acinetobacter johnsonii and was successfully applied to generate kinetic data. No substantial difference in the estimated maximum specific growth rate (lmax) values was found between the SQ method and the classical method, using absorbance at 420 nm (6Æ2 d)1). The preliminary validation tests showed their direct applicability to target enriched cultures. Conclusions: This study demonstrated the validity of the SQ method to easily quantify the concentration and to determine the growth rate of specific microorganisms present in mixed cultures. Significance and Impact of the Study: The proposed method can be directly utilized for quantification and kinetic characterization of microbial enrichments. It has the advantage of being easily applicable using simple, inexpensive equipment suitable for routine analysis |
| Source | Letters in applied microbiology 44 (6), pp. 1–6 |
| Keywords | Acinetobacter johnsoniibacterial growth kineticsFISH |
| Journal | Letters in applied microbiology |
| Editor | Blackwell Scientific for the Society for Applied Bacteriology,, Oxford, Regno Unito |
| Year | 2007 |
| Type | Articolo in rivista |
| DOI | 10.1111/j.1472-765X.2007.02121.x |
| Authors | ROSSETTI S. (1); TOMEI C. (1); BLACKALL L. (2); TANDOI V. (1) |
| Text | 42151 2007 10.1111/j.1472 765X.2007.02121.x ISI Web of Science WOS 000247227700012 Acinetobacter johnsonii bacterial growth kinetics FISH BACTERIAL GROWTH KINETICS ESTIMATION BY FLUORESCENCE IN SITU HYBRIDIZATION AND SPECTROFLUOROMETRIC QUANTIFICATION ROSSETTI S. 1 ; TOMEI C. 1 ; BLACKALL L. 2 ; TANDOI V. 1 1 Water Research Institute, IRSA CNR, Via Reno, Rome, Italy 2 Advanced Wastewater Management Centre, The University of Queensland, St Lucia, Qld, Australia Aims The aim of this study was to develop a specific and rapid method to identify and quantify relevant bacterial populations in mixed biomass by spectrofluorometric quantification SQ of whole cells hybridized with fluorescently labelled oligonucleotide probes targeting mature 16S ribosomal RNA rRNA . Probe targeting the precursor of rRNA synthesis was also employed because it was being suggested as more indicative of the activity state of the microorganisms. Methods and Results Original fluorescence in situ hybridization protocol was modified to be applied to liquid samples and the fluorescence emission from the Cy3 labelled cells was measured by spectrofluorometry. The method was calibrated on an exponentially growing cell suspension of Acinetobacter johnsonii and was successfully applied to generate kinetic data. No substantial difference in the estimated maximum specific growth rate lmax values was found between the SQ method and the classical method, using absorbance at 420 nm 6Æ2 d 1 . The preliminary validation tests showed their direct applicability to target enriched cultures. Conclusions This study demonstrated the validity of the SQ method to easily quantify the concentration and to determine the growth rate of specific microorganisms present in mixed cultures. Significance and Impact of the Study The proposed method can be directly utilized for quantification and kinetic characterization of microbial enrichments. It has the advantage of being easily applicable using simple, inexpensive equipment suitable for routine analysis 44 Bacterial growth kinetics estimation by fluorescence in situ hybridization and spectrofluorometric quantification rossetti_LAM2007.pdf Articolo in rivista Blackwell Scientific for the Society for Applied Bacteriology, 0266 8254 Letters in applied microbiology Letters in applied microbiology Lett. appl. microbiol. Letters in applied microbiology simona.rossetti ROSSETTI SIMONA mariaconcetta.tomei TOMEI MARIA CONCETTA valter.tandoi TANDOI VALTER |