Cells rely on diffusion to move metabolites and biomolecules. Diffusion is highly efficient but only over short distances. Although eukaryotic cells have broken free of diffusion-dictated constraints on cell size, most bacteria and archaea are forced to remain small. Exceptions to this rule are found among the bacterial symbionts of surgeonfish; Epulopiscium spp. are cigar-shaped cells that reach lengths in excess of 600 m. A large Epulopiscium contains thousands of times more DNA than a bacterium such as Escherichia coli, but the composition of this DNA is not well understood. Here, we present evidence that Epulopiscium contains tens of thousands of copies of its genome. Using quantitative, single-cell PCR assays targeting single-copy genes, we have determined that copy number is positively correlated with Epulopiscium cell size. Although other bacteria are known to possess multiple genomes, polyploidy of the magnitude observed in Epulopiscium is unprecedented. The arrangement of genomes around the cell periphery may permit regional responses to local stimuli, thus allowing Epulopiscium to maintain its unusually large size. Surveys of the sequences of single-copy genes (dnaA, recA, and ftsZ) revealed genetic homogeneity within a cell consistent with only a small amount (Ϸ1%) of the parental DNA being transferred to the next generation. The results also suggest that the abundance of genome copies in Epulopiscium may allow for an unstable genetic feature, a long mononucleotide tract, in an essential gene. With the evolution of extreme polyploidy and large cell size, Epulopiscium has acquired some of the advantages of eukaryotic cells.dnaA ͉ Epulopiscium ͉ large bacteria ͉ mononucleotide repeat ͉ polyploid I t is well appreciated that many eukaryotes are orders of magnitude larger than all known members of the Bacterial and Archaeal domains. Eukaryotes have broken free of constraints on cell size by the development of sophisticated nutrient uptake systems, subcellular compartmentalization, and the use of a cytoskeleton and motor proteins to transport vesicles. With the further advance of multicellularity, cell and tissue specialization have allowed eukaryotes to attain tremendous dimensions (1, 2). Until recently (3, 4), bacterial (and archaeal) cells were considered simple, displaying little subcellular organization. Although we now know that bacterial cells are also highly organized, possessing motor and cytoskeletal proteins, and even extensive intracellular membranes in some instances (5), these cells are believed to rely on diffusion to access nutrients and other metabolically important chemicals. Diffusion coefficients of small molecules may impose time constraints on metabolite flux (6) that require bacterial cells to maintain very high surface-tovolume ratios. As a result, no part of the cytoplasm is very far from the external environment, and so exchange is rapid enough to sustain metabolic processes. Most large bacteria fit this paradigm and maintain a very thin cytoplasm; they are extremely long and sle...

Aims:  Understanding factors that influence the composition of microbial populations of the digestive system of dairy cattle will be key in regulating these populations to improve animal performance. Although rumen microbes are well studied, little is known of the dynamics and role of microbial populations in the small intestine of cows. Comparisons of fingerprints of microbial populations were used to investigate the effects of gastrointestinal (GI) segment and animal on community structure. Methods and Results:  Samples from four lactating dairy cows with ruminal, duodenal and ileal cannulae were collected. Terminal‐restriction fragment length polymorphism (T‐RFLP) comparisons of small subunit rRNA genes revealed differences in microbial populations between GI segments (P < 0·05). No significant differences in either methanogen populations or microbial community profiles between animals were observed. Quantitative PCR was used to assay relative changes in methanogen numbers compared to procaryote rRNA gene numbers, and direct microscopic counts were used to enumerate total procaryote numbers of the duodenal and ileal samples. Conclusions:  T‐RFLP comparisons illustrate significant changes in microbial diversity as digesta passes from one segment to another. Direct counts indicate that microbial numbers are reduced by eight orders of magnitude from the rumen, through the abomasum, and into the duodenum (from c. 1012 to c. 3·6 × 104 cells per ml). Quantitative PCR analyses of rRNA genes indicate that methanogens are present in the duodenum and ileum. Significance and Impact of the Study:  The contribution of microbial populations of the small intestine to the nutrition and health of cattle is seldom addressed but warrants further investigation.