Quantitation of microbial cell surface heterogeneity by microelectrophoresis and electron microscopy — Application to lactobacilli after serial passaging

• Published in: Journal of microbiological methods Vol. 19; no. 4; pp. 269 - 277
• Main Authors: Geertsema-Doornbusch, G.I., Noordmans, J., Bruce, A.W., Reid, G., Khoury, A.E., van der Mei, H.C., Busscher, H.J.
• Format: Journal Article
• Language: English
• Published: Shannon Elsevier B.V 1994; Elsevier Science
• Subjects: Bacteriological methods and techniques used in bacteriology; Bacteriology; Biological and medical sciences; Cell surface heterogeneity; Cell surface structure; Fundamental and applied biological sciences. Psychology; Lactobacillus; Microbiology; Zeta potential; Cell surface heterogeneity; Zeta potential; Cell surface structure; Lactobacillus; Characterization; Heterogeneity; Gel electrophoresis; Bacteria; Lactobacillaceae; Method; Electron microscopy; Cell surface
• ISSN: 0167-7012; 1872-8359
• DOI: 10.1016/0167-7012(94)90030-2

This paper attempts to demonstrate the use of particulate microelectrophoresis as a method to quantitate subpopulations with different cell surface properties in pure, single cultures of microbial cells. To this end, two primary isolates of urogenital lactobacilli were serially passaged in liquid medium up to 50 times. In one strain, serial passaging was accompanied by the development of structural cell surface heterogeneity, in which about half of the cells possessed a thick ruthenium red/uranyl acetate stained layer observed by transmission electron microscopy on sectioned cells, whereas the other half of the cells was devoid of such a layer. Particulate microelectrophoresis with an automated image analysis system enabled determination of the zeta potentials of individual cells in a suspension, showing that in 10 mM potassium phosphate solution, pH 5.0, the primary isolate of this strain had a single zeta potential of +2.8 mV. In both p = 20 and p = 50 cultures a more negatively charged subpopulation (zeta potentials −19.7 and −18.3 mV for p = 20 and p = 50, respectively) existed next to the virtually uncharged fraction. The other strain developed a less clear structural cell surface heterogeneity after serial passaging and consequently only a small shift in zeta potentials occurred, albeit that the standard deviation over the zeta potential distribution increased from 2.3 to 3.4 mV after serial passaging. It is concluded that particulate microelectrophoresis is a good method to quantitate subpopulations with different cell surface properties and that it is less time consuming that electron microscopy. Depending on the nature of the cell surface heterogeneity, either two zeta potential distributions are measured for one cultured or the standard deviation over a single zeta potential distribution is enlarged.