Scholarly Work - Center for Biofilm Engineering
Permanent URI for this collectionhttps://scholarworks.montana.edu/handle/1/9335
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Item Optimal surface estimation and thresholding of confocal microscope images of biofilms using Beer's Law(2020-05) Parker, Albert E.; Christen, J. A.; Lorenz, Lindsey A.; Smith, Heidi J.Beer's Law explains how light attenuates into thick specimens, including thick biofilms. We use a Bayesian optimality criterion, the maximum of the posterior probability distribution, and computationally efficiently fit Beer's Law to the 3D intensity data collected from thick living biofilms by a confocal scanning laser microscope. Using this approach the top surface of the biofilm and an optimal image threshold can be estimated. Biofilm characteristics, such as bio-volumes, can be calculated from this surface. Results from the Bayesian approach are compared to other approaches including the method of maximum likelihood or simply counting bright pixels. Uncertainty quantification (i.e., error bars) can be provided for the parameters of interest. This approach is applied to confocal images of stained biofilms of a common lab strain of Pseudomonas aeruginosa, stained biofilms of Janthinobacterium isolated from the Antarctic, and biofilms of Staphylococcus aureus that have been genetically modified to fluoresce green.Item Janthinobacterium CG23_2: comparative genome analysis reveals enhanced environmental sensing and transcriptional regulation for adaptation to life in an Antarctic supraglacial stream(2019-10) Dieser, Markus; Smith, Heidi J.; Ramaraj, Thiruvarangan; Foreman, Christine M.As many bacteria detected in Antarctic environments are neither true psychrophiles nor endemic species, their proliferation in spite of environmental extremes gives rise to genome adaptations. Janthinobacterium sp. CG23_2 is a bacterial isolate from the Cotton Glacier stream, Antarctica. To understand how Janthinobacterium sp. CG23_2 has adapted to its environment, we investigated its genomic traits in comparison to genomes of 35 published Janthinobacterium species. While we hypothesized that genome shrinkage and specialization to narrow ecological niches would be energetically favorable for dwelling in an ephemeral Antarctic stream, the genome of Janthinobacterium sp. CG23_2 was on average 1.7 ± 0.6 Mb larger and predicted 1411 ± 499 more coding sequences compared to the other Janthinobacterium spp. Putatively identified horizontal gene transfer events contributed 0.92 Mb to the genome size expansion of Janthinobacterium sp. CG23_2. Genes with high copy numbers in the species-specific accessory genome of Janthinobacterium sp. CG23_2 were associated with environmental sensing, locomotion, response and transcriptional regulation, stress response, and mobile elements—functional categories which also showed molecular adaptation to cold. Our data suggest that genome plasticity and the abundant complementary genes for sensing and responding to the extracellular environment supported the adaptation of Janthinobacterium sp. CG23_2 to this extreme environment.Item Impact of hydrologic boundaries on microbial planktonic and biofilm communities in shallow terrestrial subsurface environments(2018-09) Smith, Heidi J.; Zelaya, Anna J.; De León, Kara B.; Chakraborty, R.; Elias, Dwayne A.; Hazen, Terry C.; Arkin, Adam P.; Cunningham, Alfred B.; Fields, Matthew W.Subsurface environments contain a large proportion of planetary microbial biomass and harbor diverse communities responsible for mediating biogeochemical cycles important to groundwater used by human society for consumption, irrigation, agriculture and industry. Within the saturated zone, capillary fringe and vadose zones, microorganisms can reside in two distinct phases (planktonic or biofilm), and significant differences in community composition, structure and activity between free-living and attached communities are commonly accepted. However, largely due to sampling constraints and the challenges of working with solid substrata, the contribution of each phase to subsurface processes is largely unresolved. Here, we synthesize current information on the diversity and activity of shallow freshwater subsurface habitats, discuss the challenges associated with sampling planktonic and biofilm communities across spatial, temporal and geological gradients, and discuss how biofilms may be constrained within shallow terrestrial subsurface aquifers. We suggest that merging traditional activity measurements and sequencing/-omics technologies with hydrological parameters important to sediment biofilm assembly and stability will help delineate key system parameters. Ultimately, integration will enhance our understanding of shallow subsurface ecophysiology in terms of bulk-flow through porous media and distinguish the respective activities of sessile microbial communities from more transient planktonic communities to ecosystem service and maintenance.Item Microbial growth under humic-free conditions in a supraglacial stream system on the Cotton Glacier, Antarctica(2013-07) Foreman, Christine M.; Cory, R. M.; Morris, Cindy E.; SanClements, M. D.; Smith, Heidi J.; Lisle, John T.; Miller, P. L.; Chin, Yu-Ping; McKnight, Diane M.During the austral summers of 2004 and 2009, we sampled a supraglacial stream on the Cotton Glacier, Antarctica. The stream dissolved organic matter (DOM) was low (44–48 µ M C) and lacked detectable humic fluorescence signatures. Analysis of the excitation emissions matrices (EEMs) indicated that amino-acid fluorophores dominated, consistent with DOM of microbial origin, with little humic-like fluorescence. In most aquatic ecosystems, humic DOM attenuates harmful UV radiation and its absence may represent an additional stressor influencing the microbial community. Nonetheless, the stream contained an active microbial assemblage with bacterial cell abundances from 2.94 x 104 to 4.97 x 105 cells ml-1, and bacterial production ranging from 58.8 to 293.2 ng C l-1 d-1. Chlorophyll-a concentrations ranged from 0.3 to 0.53 µ g 1-1 indicating that algal phototrophs were the probable source of the DOM. Microbial isolates produced a rainbow of pigment colors, suggesting adaptation to stress, and were similar to those from other cryogenic systems (Proteobacteria and Bacteroidetes lineages). Supraglacial streams provide an example of contemporary microbial processes on the glacier surface and a natural laboratory for studying microbial adaptation to the absence of humics.Item Draft genome sequence and description of Janthinobacterium sp. strain CG3, a psychrotolerant antarctic Supraglacial stream bacterium(2013-11) Smith, Heidi J.; Akiyama, Tatsuya; Foreman, Christine M.; Franklin, Michael J.; Woyke, Tanja; Teshima, H; Davenport, K.; Daligault, H.; Erkkila, T.; Goodwin, L. A.; Gu, W.; Xu, Yan; Chain, P. S.Here we present the draft genome sequence of Janthinobacterium sp. strain CG3, a psychrotolerant non-violacein-producing bacterium that was isolated from the Cotton Glacier supraglacial stream. The genome sequence of this organism will provide insight into the mechanisms necessary for bacteria to survive in UV-stressed icy environments.Item Draft genome sequence of a metabolically diverse Antarctic supraglacial stream organism, Polaromonas sp. strain CG9_12, determined using pacific biosciences single-molecule real-time sequencing technology(2014-12) Smith, Heidi J.; Foreman, Christine M.; Ramaraj, ThiruvaranganPolaromonas species are found in a diversity of environments and are particularly common in icy ecosystems. Polaromonas sp. strain CG9_12 is an aerobic, Gram-negative, catalase-positive, white-pigmented bacterium of the Proteobacteria phylum. Here, we present the draft genome sequence of Polaromonas sp. strain CG9_12, isolated from an Antarctic supraglacial stream.Item Archaeal ammonium oxidation coupled with bacterial nitrite oxidation in a simulated drinking water premise plumbing system(2016-01) Santillana, G. E.; Smith, Heidi J.; Burr, Mary; Camper, Anne K.Simulated copper and PVC premise plumbing reactors modeling chloramine decay were monitored for complete nitrification of 0.71 mg NH4-N L−1 ammonium to nitrate with no nitrite detected. PCR, qPCR, fluorescent in situ hybridization (FISH) and DNA sequencing were used to investigate the microbial community responsible for nitrification in the reactors' influent and biofilm on copper and PVC surfaces. No bacterial ammonium oxidizers were detected by directly targeting the bacterial amoA gene or 16S rRNA gene amplicons. FISH images indicated an archaeal population on both surfaces. Archaeal 16S rRNA and amoA gene sequences showed 98.6% and 87.6% similarity to the known archaeal ammonium oxidizer, Candidatus Nitrosotenuis uzonenis. Copy numbers of the archaeal 16S rRNA gene and archaeal amoA approximated a 1:1 ratio, suggesting that any archaea in the systems are likely to be ammonium oxidizers. Further, there was evidence for the presence of bacterial nitrite oxidizers. Copper surfaces supported fewer archaea as detected using the archaeal 16S rRNA and amoA genes. The results provide strong evidence for biofilms in a drinking water premise plumbing system composed of archaeal ammonium oxidizers and bacterial nitrite oxidizers, capable of complete oxidation of ammonium to nitrate. Since no bacterial ammonium oxidizers were found, this study adds to the growing body of research indicating an important role for archaeal ammonium oxidizers in freshwater/drinking water environments in the conversion of ammonium to nitrite.Item Genome sequence of Janthinobacterium sp. CG23_2, a violacein-producing isolate from an Antarctic supraglacial stream.(2016-01) Smith, Heidi J.; Foreman, Christine M.; Akiyama, Tatsuya; Franklin, Michael J.; Devitt, N. P.; Ramaraj, ThiruvaranganHere, we present the draft genome sequence for the violacein-producing Janthinobacterium sp. CG23_2 isolated from an Antarctic supraglacial stream. The genome is ~7.85 Mb, with a G+C content of 63.5%. The genome includes 7,247 candidate protein coding genes, which may provide insight into UV tolerance mechanisms.