Prebiotics are selectively fermented ingredients that allow specific changes in the gastrointestinal microbiota that confer health benefits to the host. However, the effects of prebiotics on the human gut microbiota are incomplete as most studies have relied on methods that fail to cover the breadth of the bacterial community. The goal of this research was to use high throughput multiplex community sequencing of 16S rDNA tags to gain a community wide perspective of the impact of prebiotic galactooligosaccharide (GOS) on the fecal microbiota of healthy human subjects. Fecal samples from eighteen healthy adults were previously obtained during a feeding trial in which each subject consumed a GOS-containing product for twelve weeks, with four increasing dosages (0, 2.5, 5, and 10 gram) of GOS. Multiplex sequencing of the 16S rDNA tags revealed that GOS induced significant compositional alterations in the fecal microbiota, principally by increasing the abundance of organisms within the Actinobacteria. Specifically, several distinct lineages of Bifidobacterium were enriched. Consumption of GOS led to five- to ten-fold increases in bifidobacteria in half of the subjects. Increases in Firmicutes were also observed, however, these changes were detectable in only a few individuals. The enrichment of bifidobacteria was generally at the expense of one group of bacteria, the Bacteroides. The responses to GOS and the magnitude of the response varied between individuals, were reversible, and were in accordance with dosage. The bifidobacteria were the only bacteria that were consistently and significantly enriched by GOS, although this substrate supported the growth of diverse colonic bacteria in mono-culture experiments. These results suggest that GOS can be used to enrich bifidobacteria in the human gastrointestinal tract with remarkable specificity, and that the bifidogenic properties of GOS that occur in vivo are caused by selective fermentation as well as by competitive interactions within the intestinal environment.

BackgroundStreptococcus thermophilus represents the only species among the streptococci that has “Generally Regarded As Safe” status and that plays an economically important role in the fermentation of yogurt and cheeses. We conducted comparative genome analysis of S. thermophilus LMD-9 to identify unique gene features as well as features that contribute to its adaptation to the dairy environment. In addition, we investigated the transcriptome response of LMD-9 during growth in milk in the presence of Lactobacillus delbrueckii ssp. bulgaricus, a companion culture in yogurt fermentation, and during lytic bacteriophage infection.ResultsThe S. thermophilus LMD-9 genome is comprised of a 1.8 Mbp circular chromosome (39.1% GC; 1,834 predicted open reading frames) and two small cryptic plasmids. Genome comparison with the previously sequenced LMG 18311 and CNRZ1066 strains revealed 114 kb of LMD-9 specific chromosomal region, including genes that encode for histidine biosynthetic pathway, a cell surface proteinase, various host defense mechanisms and a phage remnant. Interestingly, also unique to LMD-9 are genes encoding for a putative mucus-binding protein, a peptide transporter, and exopolysaccharide biosynthetic proteins that have close orthologs in human intestinal microorganisms. LMD-9 harbors a large number of pseudogenes (13% of ORFeome), indicating that like LMG 18311 and CNRZ1066, LMD-9 has also undergone major reductive evolution, with the loss of carbohydrate metabolic genes and virulence genes found in their streptococcal counterparts. Functional genome distribution analysis of ORFeomes among streptococci showed that all three S. thermophilus strains formed a distinct functional cluster, further establishing their specialized adaptation to the nutrient-rich milk niche. An upregulation of CRISPR1 expression in LMD-9 during lytic bacteriophage DT1 infection suggests its protective role against phage invasion. When co-cultured with L. bulgaricus, LMD-9 overexpressed genes involved in amino acid transport and metabolism as well as DNA replication.ConclusionsThe genome of S. thermophilus LMD-9 is shaped by its domestication in the dairy environment, with gene features that conferred rapid growth in milk, stress response mechanisms and host defense systems that are relevant to its industrial applications. The presence of a unique exopolysaccharide gene cluster and cell surface protein orthologs commonly associated with probiotic functionality revealed potential probiotic applications of LMD-9.

We hypothesize that a reduced capacity to withstand or repair cellular damage from ultraviolet radiation may be present in caveadapted microorganisms that never experience such conditions. However, a small number of previous studies have shown that some subsurface bacteria do not show greater sensitivity to ultraviolet radiation (UVR) than surface bacteria. To estimate UVR sensitivity in cave bacteria, bacterial isolates were collected from Carlsbad Cavern, New Mexico, U.S.A., and percent survival following exposure to various UVC and UVA radiation doses was determined. Cave bacteria from Left Hand Tunnel in Carlsbad Cavern and surface bacteria from soil and rocks above Carlsbad Cavern were grown on low and high nutrient media then exposed to 0, 10,000 and 20,000 μWs/ cm 2 of UVR in a laboratory biological safety cabinet. Incubations were conducted at 15°C or 37ºC, in accordance with the isolates' natural temperature environments. In addition, DNA repair capacity was estimated by exposing the organisms to various doses of UVC radiation and measuring survivability. Gram status and pigmentation also were determined. Results showed that most of the cave isolates were more sensitive to UVR than the surface isolates, but survivability data suggest that cave microbes retain some of their capacity to repair UV-induced DNA damage. Selection appears to have favored bacteria that can survive in this low nutrient environment, while not maintaining (or paying the cost of maintaining) unneeded traits such as UVR resistance. Cave bacteria appear to have maintained DNA repair capacity, most likely because of the need to repair damage to their DNA from other environmental stressors found in caves.Keywords: Ultraviolet radiation sensitivity, cave-adaptation, bacteria, subsurface, caves unable to show a correlation between natural levels of radiation exposure and species resistance (Nasim & James, 1978;Arrage et al., 1993a).UV radiation (UVR) at wavelengths shorter than 400 nm is absorbed by DNA and can cause cyclobutane dimer formation, interstrand crosslinking, and other direct and indirect damage to DNA (Nasim & James, 1978;Miller et al., 1999). In addition to enzymatic DNA repair mechanisms, microbes have evolved to survive UV exposure through other molecular and structural protection and avoidance methods. However, whether these additional methods actually protect the bacterial DNA is widely debated (Mathews & Sistrom, 1959; Dworkin & Stanley, 2006;Gascon et al., 1995; Lewis et al., 1973; Singer & Ames, 1970; Nasium & James, 1978;Cockell, 1998;Arrage et al., 1993a). Iron compounds, sand, desert crust, rock, water, sulfur and sodium chloride have all been found to be natural UVR shields for bacteria (Cockell, 1998;Rothschild & Giver, 2003). A well-documented predominance of pigmentation in UVR-resistant species of bacteria suggests the use of carotenoids, yellow, orange and red cellular pigments, as UVR screening agents (Mathews & Sistrom, 1959; Dworkin & Stanley, 2006). UVR screening agents also provide protection from ox...