Subdivision of the genus Klebsiella on the basis of DNA-DNA membrane filter hybridization Public Deposited

http://ir.library.oregonstate.edu/concern/graduate_thesis_or_dissertations/7h149s49p

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  • The molecular relationship of 39 biochemically identified strains of Klebsiella pneumoniae was determined by DNA-DNA hybridization experiments. Cultures examined were isolated from vegetables, pulp mills, drinking water and clinical (human and animal) habitats. These organisms produced reassociation values of 7% to 100% with ATCC 13882 K. pneumoniae as the labeled reference indicating a high degree of heterogeneity within the species. After dividing cultures into three groups on the basis of hybridization values, it was observed that isolates exhibiting low values to ATCC 13882 were most often from 'environmental' habitats in contrast to those highly related to ATCC 13882 which were predominantly from clinical origins. No discernable pattern was observed for those cultures producing intermediate values to ATCC 13882, i.e., cultures were isolated from environmental and clinical sources. Additional hybridization experiments were carried out using a vegetable isolate (V-236) and K. pneumoniae biotype oxytoca (ATTC 13182) as labeled references. Values obtained for these two organisms correlated to a high degree with the two low hybridization groups, relative to ATCC 13882, established above. Cultures which were highly related to V-236 exhibited low molecular relatedness (less than 40%) to ATCC 13882 while cultures highly related to ATCC 13182 exhibited intermediate relatedness (40% to 75%). Furthermore, the internal homogeneity of each group, ATCC 13882, ATCC 13182 and V-236, is sufficient to recommend the establishment of three independent species within the genus Klebsiella corresponding to each group. Although biochemically similar, each group was found to have phenotypically distinct reaction patterns to four tests. These tests include indole production (I), pectin liquefaction (Pc), fecal coliform response (FC) and growth at 10°C (10°). The most frequent reaction pattern associated with each group is I-, Pc-, FC+, 10°- for ATCC 13882, I+, Pc+, FC-, 100+ for ATCC 13182 and I-, Pc-, FC-, 10°+ for V-236. The molecular, phenotypic and environmental relationships of these three groups is of public health significance. This is due to the frequency with which Klebsiella exhibiting a FC+ reaction are isolated from a clinical environment and, in general, the molecular relationship between organisms of the group which are FC+ and FC-. In other words, by following the above recommendations, essentially all FC- Klebsiella would be removed from the species K. pneumoniae thereby leaving only those organisms which are FC+ and clearly associated with clinical environments.
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