Proceedings Volume 2010 (format .pdf) - SimpBTH

As one can see in figure 4, the highest active cell densities were found in M1 andthe lowest in M3 (control) probably because of the extra nutrients in M1 (gasolineand ammonium acetate); however, the fastest increase in the number of active cellswas found in M2 (0.3x10 6 cells/mL at 2 hours and 1.2x10 6 cells/mL at 8 hours ofincubation).Taking into account only the cells which increased their length (fig.4) to calculatethe average cell length of these cells, the following values have been obtained:5.82±1.20 for M1; 7.71±0.54 for M2 and 3.15±0.86 for M3. As expected, thesevalues are larger than average values obtained for all cells, both growing and nongrowing (fig.2).CONCLUSIONSIn this paper the original direct viable counts method (Kogure et al., 1978) is usedto count cells that are capable of division in the presence of gasoline, and for thefirst time, up to our best knowledge, the microscopic images obtained with thismethod were treated by special software (Image J and Cell C) for automatedimage analysis which increases the accuracy of the measurements.The presented methods including image analysis systems were performed forcounting and estimating the length of bacteria, but does not totally exclude theneed for double blind validation of both manual microscope analyses andautomated image analysis.REFERENCES1. Ardelean, I., Ghiţă S., Sarchizian, I. (2009a): Isolation of oxygenic phototrophic and oxicheterotrophic bacteria with potential for gasoline consumption. Proceedings of the 2 nd InternationalSymposium “New Research in Biotechnology”, serie F, pp. 278-287.2. Ardelean, I., Ghiţă, S., Sarchizian, I. (2009b): Epifluorescent method for quantification ofplanktonic marine prokaryotes. Proceedings of the 2 nd International Symposium “New Research inBiotechnology”, serie F, pp. 288-296.3. Bakermans, C. & Madsen, E.L. (2000): Use of substrate responsive-direct viable counts to visualizenaphthalene degrading bacteria in a coal tar-contaminated groundwater microbial community. J.Microbiol. Meth. Vol. 43: 81-90.4. Barcina, I., Arana, I., Santoram, P., Iriberri, J., Egca, L. (1995): Direct viable count gram positiveand gram negative bacteria using ciproflocacin as inhibitor of cellular division. J. Microbiol.Methods. Vol. 22: 139-150.5. Baudart, J., Coallier, J., Laurent, P., Prevost, M. (2002): Rapid and sensitive enumeration of viablediluted cells of members of the family enterobacteriaceae in freshwater and drinking water. Appl.Environ. Microbiol. Vol. 68: 5057-5063.6. Congestri, R., Capucci, E., Albertano, P. (2003): Morphometric variability of the genus Nodularia(Cyanobacteria) in Baltic natural communities. Aquat. Microb. Ecol. Vol. 32: 251-259.7. Embleton, K.V., Gibson, C.E., Heaney, S.I. (2003): Automated counting of phytoplankton bypattern recognition: a comparison with a manual counting method. J. Plankt. Res. Vol. 25: 669-681.8. Estep, K.W. & Macintyre, F. (1989): Counting, sizing, and identification of algae using imageanalysis. Sarsia. Vol. 74: 261-268.9. Fry, J.C. (1990): Direct methods and biomass estimation. Methods in Microbiology. Vol. 22: 51-67.211