Phenetic and Phylogenetic Analysis of Kocuria palustris and Kocuria rhizophila Strains isolated from Healthy and Thalassemia Persons

Objective: The normal habitat of Kocuria palustris and K. rhizophila include mammalian skin, soil and rhizoplane. The aim of this study is to find the relatedness among several strains of K. palustris and K. rhizophila isolated from healthy and thalassemia patients according to the biochemical tests, make phylogenetic analysis to construct a database for whole cell protein band profile of these species. Methods: Ninety samples were collected from healthy and thalassemia patients skin in April of 2013, seventeen samples were revealed bacterial isolates. The diagnosis was performed using conventional biochemical tests , eight of them analyzed according to their 16S rRNA gene sequence and used as reference to confirm the diagnosis of other isolates depending on phenetic and protein bands clustering patterns. Three different software used , IBM SPSS v.19, MEGA 5.22 and CLIQS v.1 in dendrogram building and interpreting the results. Results: Seventeen strains of the genus Kocuria were isolated from 90 sample collected from human skin. These strains appeared as coccoid Gram positive cells and had smooth yellow colonies, slightly convex. Three strains were related to K. palustris and fourteen strains were related to K. rhizophila. The morphological dendrogram built upon 76 phenetic characters, and divided to three clusters with similarity ratio of 94-99%. Eight strains were analyzed for their 16S ribosomal RNA gene sequences and compared with NCBI by using BLAST which gave 94-97 % similarity with reference strains. The whole cell protein profile was analyzed by CLIQS v.1, similarity ratio ranged from 29-100% among strains in dendrogram. Analysis of amino acids composition in cell wall using paper chromatography showed four amino acids, alanine, glutamic acid, glycine and lysine. Conclusion: The study showed good relatedness among strains of K. palustris and K. rhizophila according to morphological and biochemical tests and phylogenetic analysis, but low similarity according to whole cell protein bands profile.