Kefir Kommunity

1. S. Dellis J. Kidd M. Martin P. Worthington What is the microbial community composition of kefir grains? Kefir Kommunity Is the community static? Is the community dynamic? Kefir Grain Variables: Normal Cultivation After Cultivation Shifts Geography Sources Isolate LAB & Yeast Isolate DNA Label LAB/Yeast with GFP 16SrRNA/18SrRNA primers Sample Tree Construction: Genbank search for “kefir” produced 75 “hits”, which were refined to select the six sequences identified as Lactobacillus kefiri. Five of these six were >220 bases in length and were subjected to a CLUSTALW alignment. Abstract: The fermented probiotic milk beverage, kefir, is made by grains consisting of a consortium of microbes (including lactic acid bacteria and yeasts) surrounded by an exopolysaccharide matrix. The members of this consortium have not been fully characterized by culture-independent methodologies, nor has the population dynamics of these kefir grains been determined. Using culture-independent methodology, we propose to investigate the population structure and possible dynamics of this microbiological consortium under several conditions: from isolate to isolate locally, from isolates at different geographical locations, and before and after changes in cultivation conditions (moving from milk to grape juice or sugar-water). This approach should reveal the level of population diversity among the residents of active fermenting grains, and how the Clone Library Isolate LAB & Yeast from non-Kefir source Transformation Isolate DNA Sequencing BLASTRibosomal Database Project Locate GFP in Kefir grains Phylogenies population structure is altered by changing culture conditions. In addition, we propose to investigate if “non-kefir associated” lactic acid bacteria can become incorporated into kefir grains during culture, using GFP labeling to reveal “microgeographic” aspects of this process, if it occurs. Finally, we will develop a “virtual meeting place” for investigators interested in this phenomenon: “Kamp Kefir.” This site will contain sequence data from experiments such as we describe carried out by as many investigators as have interest in this approach, as well as information about kefir and data pertaining to this interesting system. The resident data will serve as an excellent teaching tool for students, particularly students interested in bioinformatics, metagenomics, and related technologies, at institutions that do not have active research programs.