The significant role of the polysaccharide structure on swarming has been revealed in previous studies (Toguchi et al., 2000; Inoue et al., 2007).

To support this www.selleckchem.com/products/azd-1208.html point, swarming cells of C. freundii were observed to be more hydrophilic compared with vegetative cells (0.961 for swarming cells; 0.814 for vegetative cells, P<0.05; Fig. S2) in this work. In swarming colonies, C. freundii cells moved actively. However, the cells at the periphery of colonies were less active due to the decreased moisture capacity in areas where the cells moved out and back occasionally, or were pushed by the actively moving cells in the central region. As a result, the edge of colonies expanded outward continuously. Citrobacter freundii did not display alternating cycles of swarming and consolidation during the development of swarming colonies as in P. mirabilis.

Once differentiation occurred at the inoculation site, swarming cells spread continuously, until they occupied the entire agar surface. Even when inoculated onto a large plate with a diameter over 20 cm, alternating cycles of swarming and consolidation were not observed. Transposon mutagenesis involving the use of Mini-Tn5 on a suicide plasmid pUT was carried out to further learn more understand the genetic determinants of swarming motility in C. freundii. A total of 85 swarming-defective mutants were screened from approximately 6000 transconjugants; of the 85 mutants, 53 were defective in both swimming and swarming. The remaining 32 mutants were defective in swarming but not swimming. The mutants with normal swimming pattern were subjected to further sequence analysis to determine the insertionally mutated gene. Given that swarming is dependent on functional flagella, as demonstrated in previous studies, of the 53 swimming-defective mutants, only five randomly selected mutants were further subjected to sequence analysis. As a whole, sequences produced valid results with only four exceptions

(CF407, CF415, CF701, and CF711). In most cases, the most similar genes obtained through the homology searches usually belonged to Citrobacter koseri ATCC BAA-895, a species of Citrobacter with complete genome sequence information. The results of the homology searches are listed in Tables 1 and 2 and are also described next in the following two sections on genes that have been previously characterized in other species and those first identified in this study. As many as 16 swarming-related genes identified in our study have already been characterized previously in other species. The underlying causes for the defective swarming motility of the mutants are listed in Table 1. However, some of them are worthy of further discussion. As expected, flhD, motA, and motB mutants were identified among the five mutants found to be defective in both swarming and swimming motilities.