Según el trabajo que he realizado hasta ahora he obtenido 2(algo de progreso, he logrado al menos 10% de los objetivos propuestos). Esto se debe a que el trabajo que estoy realizando conlleva de mucho tiempo y trabajo lo cual yo pensaba que era lo contrario, para no atrasar el trabajo he trabajado algunas horas extras. Dado a esto no voy a presentar mi trabajo en el meeting en Mayagüez, en este caso presentare el trabajo que realice el semestre pasado.
Comparison of physiological conditions during biofilm formation of Bacillus subtilis eftA mutant incapable of calcite formation
Crystal formation in bacteria grown on precipitation medium is a phenomenon with applications ranging from the restoration of historic monuments to the formation of kidney stones in humans. Even tough it is an important phenomenon not much is known about the physiological conditions of the bacteria growing within the biofilms that lead to crystal formation. To gain a better understanding of the physiology of Bacillus subtilis biofilm grown using precipitation media we have compared the growth of wild type strains and the eftA mutant which is unable to precipitate calcite. To learn more about the pH within the biofilm microenvironment we added two pH indicators (bromothymol blue and phenol red) to the growth media. After an incubation of one week at 37ºC distinct colorimetric changes were found that clearly indicated biofilm of eftA mutant were more acidic in comparison with the wild type strain. An independent study has shown that etfA is probably a subunit of a flavoprotein. Consequently, we used a commercial kit to compare the NADH/NAD ratio within B. subtilis wild type and eftA biofilm grown on precipitation media. Our preliminary results have shown that biofilm from the eftA mutant exhibited an increase at least 35 times more in their NADH/NAD ratio when compared to WT. Our next step is to determine pH changes during biofilm formation using microelectrodes and the use of an inhibitor of the electron transport chain to specifically confirm our findings.
