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COLOR PLATES

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Image of COLOR PLATE 1 (chapter 1)

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COLOR PLATE 1 (chapter 1)

Diagrammatic representation of the repetitive nature of dental plaque development between typical daily oral hygiene procedures. Initial colonizers recognize receptors in the salivary pellicle that coats the enamel surface of teeth and are detected within minutes after oral hygiene measures. (), (), (), (), and () bind to salivary receptors or coadhere to already-attached bacteria. Early colonizers spp. (), spp. (), spp. (), spp. (), spp. (), and spp. () typically appear after initial colonizers and before plaque samples are taken at 4 h after oral hygiene procedures. All species coaggregate with initial colonizers. Growth of bacteria is depicted by increased numbers of shapes representing streptococci and actinomyces, but growth is not limited to any particular species in the developing dental plaque biofilm. Middle colonizers (), spp. (), and () coaggregate with initial and early colonizers and typically appear at between 4 and 8 h. and other initial and early colonizers continue to grow. Late colonizers (), (), (), and spp. () coaggregate with and typically appear after 8 h. Growth of the biofilm flourishes, and additional species may be added to the biofilm before the biofilm is removed by oral hygiene procedures. The temporal nature of colonization of enamel is based on in vivo colonization reported in references 25, 27, 28, 82, 94, 95, 97, 98, and 119, and coaggregations are summarized in references 65, 71, and 72. The colors of the depicted species are those of the yellow, blue, purple, green, orange, and red complexes in the work of Socransky et al. (119).

Citation: Kolenbrander P. 2011. COLOR PLATES, In Oral Microbial Communities. ASM Press, Washington, DC.
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Image of COLOR PLATE 2 (chapter 3)

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COLOR PLATE 2 (chapter 3)

Comparisons between proteins predicted from streptococcal genomes. (A) Five-way comparison between five strains of Strain names are indicated at the head of each oval, followed by the total number of predicted proteins for that strain between parentheses. Each strain is coded with a letter (e.g., B for strain SK137). These letter codes are used to indicate the actual number of proteins that a given strain contributes to any given intersection of the Venn diagram. For instance, the middle section shared by all five strains lists five sets of proteins. Numbers vary among strains because of differing numbers of paralogs in any given strain. (B) Comparison of strains considered as a group (see text) to the individual genomes of and Letter codes from panel A are kept the same. (C) Comparison of grouped strains to grouped strains ( = 11). Given the larger number of genomes, only averages across all genomes are reported instead of gene counts per individual genome.

Citation: Kolenbrander P. 2011. COLOR PLATES, In Oral Microbial Communities. ASM Press, Washington, DC.
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Image of COLOR PLATE 3 (chapter 4)

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COLOR PLATE 3 (chapter 4)

Competence regulon in In step 1, the gene encodes a precursor peptide. The peptide is cleaved and exported by unknown proteins in step 2. The mature ComC, termed CSP, accumulates in the extracellular environment. CSP is sensed by the ComD sensor kinase, which responds by phosphorylating the ComE response regulator in step 3. Phosphorylated ComE activates expression of the operon and in step 4. ComX serves as an alternative sigma factor, directing the expression of “late genes” in step 5 required for uptake and incorporation of foreign DNA (step 6) and other functions.

Citation: Kolenbrander P. 2011. COLOR PLATES, In Oral Microbial Communities. ASM Press, Washington, DC.
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Image of COLOR PLATE 4 (chapter 6)

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COLOR PLATE 4 (chapter 6)

In silico reconstruction of ATCC 10790 carbohydrate metabolism glycolysis/gluconeogenesis pathway with KAAS, an automatic genome annotation and pathway reconstruction server (41). An interactive version of the pathway is available at http://www.kegg.jp/kegg/atlas, KEGG global metabolic maps, “edit,” (vpr). A green box identifies the presence of a gene or enzyme; a white box indicates that no homolog was found. The presence of phosphoglyceromutase (EC 5.4.2.1) and the absences of a hexokinase (EC 2.7.1.1), noted by Rogosa et al. (55), and a glucose phosphotransferase system (PTS permease) (EC 2.7.1.69), noted by Winter and Delwiche (61), are indicated with red arrows. (©2010 Kanehisa Laboratories, used with permission.)

Citation: Kolenbrander P. 2011. COLOR PLATES, In Oral Microbial Communities. ASM Press, Washington, DC.
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Image of COLOR PLATE 5 (chapter 6)

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COLOR PLATE 5 (chapter 6)

Confocal micrographs of immunofluorescence- and fluorescence in situ hybridization (FISH)-treated 8-h plaque on enamel showing cells reactive with the VEI488 FISH probe for veillonella 16S rRNA (green) (A), RPS-bearing streptococci reactive with anti- 34 RPS (red) (B), and an overlay of panels A and B showing juxtaposition of veillonellae and RPS-bearing streptococci (C). All images are maximum-projection images. Bar, 40 μm. Reproduced with permission from the work of Chalmers et al. (8).

Citation: Kolenbrander P. 2011. COLOR PLATES, In Oral Microbial Communities. ASM Press, Washington, DC.
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Image of COLOR PLATE 6 (chapter 7)

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COLOR PLATE 6 (chapter 7)

Spatiotemporal model of oral bacterial colonization, showing recognition of salivary pellicle receptors by early-colonizing bacteria and coaggregations between early colonizers, fusobacteria, and late colonizers of the tooth surface. Several kinds of coaggregations are shown as complementary sets of symbols of different shapes. One set is depicted in the box at the top. Proposed adhesins (symbols with a stem) represent cell surface components that are heat inactivated and protease sensitive; their complementary receptors (symbols without a stem) are unaffected by heat or protease. Identical symbols represent components that are functionally similar but may not be structurally identical. Rectangular symbols represent galactose-inhibitable coaggregations. Other symbols represent components that have no known inhibitor. Fusobacterial adhesins involved in galactose-inhibitable coaggregations are highlighted in red, FadA in blue, and RadD in green. The bacterial strains shown are , and spp. Modified from (28) with permission.

Citation: Kolenbrander P. 2011. COLOR PLATES, In Oral Microbial Communities. ASM Press, Washington, DC.
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Image of COLOR PLATE 7 (chapter 7)

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COLOR PLATE 7 (chapter 7)

Method used by the Roche 454 sequencer to amplify single-stranded DNA copies from a fragment library on agarose beads. sstDNA, single-stranded template DNA. Reprinted from the (33) with permission.

Citation: Kolenbrander P. 2011. COLOR PLATES, In Oral Microbial Communities. ASM Press, Washington, DC.
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Image of COLOR PLATE 8 (chapter 12)

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COLOR PLATE 8 (chapter 12)

biofilm formation. Following deposition onto a salivacoated glass surface, cells begin to develop pseudohyphae after 2 h at 37°C (A), with formation of true hyphae clearly visible at 5 h (B). biofilms produced under flow at 24 h show a base layer composed principally of yeast morphology cells (blastospores) and an upper layer consisting of developing hyphae, as visualized by CSLM (C). In mixedspecies biofilms of and , the streptococci can be visualized by scanning electron microscopy as being well distributed among the hyphae, but also in close association with some hyphae (D). Scale bars: panels A and B, 20 μm; panel D, 10 μm. Images in panels A, B, and C were supplied by Lindsay Dutton, University of Bristol. The image in panel D is reproduced with permission from reference 54.

Citation: Kolenbrander P. 2011. COLOR PLATES, In Oral Microbial Communities. ASM Press, Washington, DC.
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Image of COLOR PLATE 9 (chapter 12)

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COLOR PLATE 9 (chapter 12)

Interactions of , and with (A) cells (green) render hyphae nonviable, shown by yellow and orange vacuoles detected by yeast live/dead stain (bar = 10 μm); (B), the presence of has no effect upon hyphal cell viability, as shown by red-stained vacuoles (bar = 10 μm); (C), adherence of fluorescein isothiocyanate-labeled Col cells (blue fluorescence) more or less exclusively to hyphae (bar = 20 μm); (D), CSLM image of fluorescein isothiocyanate-labeled cells (green) adhering in clusters and cross-bridging hyphae (blue, stained with calcofluor) (bar = 5 μm).

Citation: Kolenbrander P. 2011. COLOR PLATES, In Oral Microbial Communities. ASM Press, Washington, DC.
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Image of COLOR PLATE 10 (chapter 14)

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COLOR PLATE 10 (chapter 14)

One-day-infected primary gingival epithelial cells (actin, red; nuclei, blue) harboring a high number of intracellular bacteria (green) undergo successful mitosis, as determined by confocal scanning fluorescence microscopy. White arrows indicate the division of the parent nucleus into two daughter nuclei. Bar, 10 μm. Reprinted from (53) with permission of the publisher.

Citation: Kolenbrander P. 2011. COLOR PLATES, In Oral Microbial Communities. ASM Press, Washington, DC.
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Image of COLOR PLATE 11 (chapter 14)

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COLOR PLATE 11 (chapter 14)

Intercellular translocation of through actin fibers at 1 day postinfection. Panel A represents three independent experiments to visualize the localization of 33277 and the organization of actin filaments using immunofluorescence microscopy. Primary gingival epithelial cells were fixed and stained with phalloidintetramethyl rhodamine isocyanate (red) and anti- antibody plus Oregon green 488-conjugated secondary antibody to visualize the intracellular (green). 4′,6-Diamidino-2-phenylindole (DAPI) (blue) was used in staining the nucleus to determine the localization of in the cytoplasm. The white arrow indicates actin projections and bacterial translocation between the host cells. spread is visualized by fluorescent microscopy. Panel B represents three independent experiments to visualize the new infection in the previously uninfected cells (green) after coincubation with the previously infected (blue) cells. The blue cells that were infected with 33277 (red) for 24 h and cocultured with the uninfected green cells for 24 h showed transmission of 33277 to newly infected cells (green), which displayed red-labeled in their cytosol. Bar, 10 μm. Reprinted from (56) with permission of the publisher.

Citation: Kolenbrander P. 2011. COLOR PLATES, In Oral Microbial Communities. ASM Press, Washington, DC.
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Image of COLOR PLATE 12 (chapter 15)

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COLOR PLATE 12 (chapter 15)

Crystal structure of mFadA. (a) Ribbon diagram of mFadA. Leucine residues are shown in ball-and-stick representation. (b) Leucine and tyrosine residues in the vicinity of the hairpin loop. The hydroxyl group of tyrosine is shown in red. (c) A trimer of mFadA formed by the interaction of the monomer in the asymmetric unit with neighboring molecules related by a simple translation along the crystallographic b axis. Leucines and residues of the hairpin loop are shown in ball-and-stick representation. Reproduced from (60) with permission.

Citation: Kolenbrander P. 2011. COLOR PLATES, In Oral Microbial Communities. ASM Press, Washington, DC.
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Image of COLOR PLATE 13 (chapter 15)

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COLOR PLATE 13 (chapter 15)

Proposed model of FadA filament formation. mFadA is secreted across the inner membrane and self-assembles in the periplasm. The green dots represent the leucine chain linking the monomers. As more molecules of mFadA are synthesized and secreted across the inner membrane, the chain elongates and extends across the outer membrane. When elongation of the filament stops, pre-FadA is incorporated to the base. It is envisaged that the N-terminal part of pre-FadA forms a short helix connected to the mFadA helix via a hairpin to form interhelical contacts presumably mediated via leucine residues on both helices. This N-terminal helical hairpin (red ribbon) could serve as an anchor in the inner membrane. Only one filament is shown, but several such filaments may interact with each other and form a bundle.

Citation: Kolenbrander P. 2011. COLOR PLATES, In Oral Microbial Communities. ASM Press, Washington, DC.
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Image of COLOR PLATE 14 (chapter 18)

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COLOR PLATE 14 (chapter 18)

The structural similarity of LuxP bound to -THMF-borate (A) and LsrB bound to -THMF (B) is evident from their three-dimensional structures. However, amino acid residues that interact with the borate diester of -THMF-borate are not conserved in LsrB, suggesting that LsrB will not interact efficiently with -THMF-borate (see text). The arrows indicate the substrate-binding pocket for each protein.

Citation: Kolenbrander P. 2011. COLOR PLATES, In Oral Microbial Communities. ASM Press, Washington, DC.
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Image of COLOR PLATE 15 (chapter 19)

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COLOR PLATE 15 (chapter 19)

In situ interactions between and oral streptococci in nascent dental plaque. Clean enamel chips were held in the mouth of a volunteer for 4 h (A) or 8 h (B) using a metal clip. Bacteria were detected with fluorescently labeled antibodies against streptococcal receptor polysaccharides (red), type 1 fimbriae (blue), and type 2 fimbriae (green). Interactions between streptococcal receptor polysaccharides and type 2 fimbriae mediate coaggregation in vitro, and these molecules colocalize in situ (line with large arrowhead). organisms bearing type 1 fimbriae are also commonly juxtaposed to streptococcal cells (lines with small arrowheads). Images were kindly provided by R. J. Palmer, Jr.

Citation: Kolenbrander P. 2011. COLOR PLATES, In Oral Microbial Communities. ASM Press, Washington, DC.
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Image of COLOR PLATE 16 (chapter 19)

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COLOR PLATE 16 (chapter 19)

Coaggregation between DL1 and MG1. Coaggregation was induced in vitro by vigorously mixing dense suspensions of and cells and appears in a test tube as macroscopic clumps, with clarification of the surrounding liquid (bottom panel). Cultures were stained with fluorescently labeled antibodies against whole cells (green) and whole cells (orange) and visualized by microscopy. and appear evenly distributed throughout clumps. Omitting the vigorous (vortex) mixing results in a coculture that initially contains few large coaggregates (middle panel). An evenly turbid monoculture is shown for comparison (top panel).

Citation: Kolenbrander P. 2011. COLOR PLATES, In Oral Microbial Communities. ASM Press, Washington, DC.
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Image of COLOR PLATE 17 (chapter 21)

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COLOR PLATE 17 (chapter 21)

Schematic (not to scale) model of initial events leading to heterotypic biofilm community development on the supragingival tooth surface. (circles) is a pioneer colonizer, and cells attach to the salivacoated tooth surface. produces multiple adhesins, many of which have cognate salivary receptors; for simplicity only SspA/B is shown. Initial localization of (rods) is mediated by interaction of FimA on the porphyromonad surface with glyceraldehyde-3-phosphate dehydrogenase (GAPDH) on the streptococcal surface. Higher-affinity binding occurs through engagement of Mfa with SspA/B. This interaction initiates a signal transduction event that modulates the transcriptome. The resulting phenotypic adaptation of , along with the production of signaling molecules such as AI-2, is necessary for the recruitment of additional cells from the planktonic phase and the initiation of community development.

Citation: Kolenbrander P. 2011. COLOR PLATES, In Oral Microbial Communities. ASM Press, Washington, DC.
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Image of COLOR PLATE 18 (chapter 21)

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COLOR PLATE 18 (chapter 21)

Sequence and predicted structural domain features of the BAR binding domain of SspB (surface of ) that interacts with Mfa (minor fimbrial protein of ). H, potential hydrophobic region, –, potential electrostatic region.

Citation: Kolenbrander P. 2011. COLOR PLATES, In Oral Microbial Communities. ASM Press, Washington, DC.
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Image of COLOR PLATE 19 (chapter 23)

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COLOR PLATE 19 (chapter 23)

Confocal micrographs and corresponding DPD (chemically synthesized AI-2) concentration-dependent changes in biovolumes of two-species biofilms containing 34 (green bars and green cells) with T14V (red bars and red cells) after 22 h of growth at 37°C in saliva-fed flow cells. The 34 wild type with T14V control is shown at the left. Biofilm cells were labeled with species-specific polyclonal antibodies conjugated to Alexa Fluor 488 ( cells) and Alexa Fluor 633 ( cells). Image taken from the work of Rickard et al. (49).

Citation: Kolenbrander P. 2011. COLOR PLATES, In Oral Microbial Communities. ASM Press, Washington, DC.
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Image of COLOR PLATE 20 (chapter 23)

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COLOR PLATE 20 (chapter 23)

(A) Photograph of a Sorbarod housed in a 3-ml syringe with a needle which delivers saliva through the plunger septum at the top of the unit. The system is designed to be a drip reactor with a bed of cellulose acetate fibers (the Sorbarod) upon which bacteria attach and form biofilms. (B) CLSM image taken of a 48-h Sorbarod biofilm that was coinoculated with 34 (green cells) and T14V (red cells). Clusters of interdigitated biofilm aggregates attach to Sorbarod fibers (shown in blue). (C) CLSM image taken of a 48-h Sorbarod biofilm that was coinoculated with 34 mutant (green cells) and T14V (red cells). Cells are mostly attached as individuals to Sorbarod fibers (shown in blue). CLSM images were generated by labeling Sorbarod fibers with calcofluor, and cells of each species were labeled with species-specific polyclonal antibodies conjugated to Alexa Fluor 488 ( 34 cells) and Alexa Fluor 633 ( T14V cells). Bar, 30 μm.

Citation: Kolenbrander P. 2011. COLOR PLATES, In Oral Microbial Communities. ASM Press, Washington, DC.
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Image of COLOR PLATE 21 (chapter 25)

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COLOR PLATE 21 (chapter 25)

Organization of multispecies microbial communities in situ within 8-h-old dental plaque showing cell-cell interweaving among at least three types of cells. General nucleic acid stain Syto 59 stains all cells (blue). Red-purple cells are reactive with an antibody against a streptococcal receptor polysaccharide involved in cell-cell recognition (coaggregation). Green cells are reactive with an antibody against many streptococcal strains known to be coaggregation partners of the receptor polysaccharide-bearing cells. Heterogeneity of antireceptor polysaccharide-reactive cells (purple) as well as antireceptor polysaccharide-unreactive streptococci (green) within single colonies is apparent and illustrates streptococcus-streptococcus intrageneric coaggregation. Multispecies communities (purple-, green-, and blue-stained cells), rather than clonal growth, characterize initial colonization of human enamel. Bar marker, 10 μm. Images are from reference 36.

Citation: Kolenbrander P. 2011. COLOR PLATES, In Oral Microbial Communities. ASM Press, Washington, DC.
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Image of COLOR PLATE 22 (chapter 25)

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COLOR PLATE 22 (chapter 25)

Representative confocal micrographs of biofilms from three-species-inoculated flow cells. Multispecies communities at 4 h (left column) and 18 h (right column) show intimate interaction of a sp. (blue) with (green) and (red) (top panels) and of a sp. (blue) with (green) and (red) (bottom panels). Images are from reference 39.

Citation: Kolenbrander P. 2011. COLOR PLATES, In Oral Microbial Communities. ASM Press, Washington, DC.
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