Mechanical damage to Gram-negative bacteria by surface plating with the Drigalski-spatula technique

Colony counting by spreading bacterial suspensions on plating media by various techniques is of general concern. Comparative studies between hand plating (Drigalski-spatula technique) for different time intervals and spiral plating resulted in significant differences in colony counts. Lower counts of Gram-negative bacteria were obtained by using hand plating for more than 10 s, compared with short time hand plating (5 s) or spiral plating. Colony counting of Gram-positive bacteria showed no differences between both techniques. Further characterisation of Escherichia coli cells spread with the Drigalski-spatula technique by electron microscopy revealed a large number of damaged cells compared to control samples. The data clearly shows that the mechanical forces during hand plating are sufficient to damage E. coli cells.