Center for Biofilm Engineering (CBE)

Permanent URI for this communityhttps://scholarworks.montana.edu/handle/1/9334

At the Center for Biofilm Engineering (CBE), multidisciplinary research teams develop beneficial uses for microbial biofilms and find solutions to industrially relevant biofilm problems. The CBE was established at Montana State University, Bozeman, in 1990 as a National Science Foundation Engineering Research Center. As part of the MSU College of Engineering, the CBE gives students a chance to get a head start on their careers by working on research teams led by world-recognized leaders in the biofilm field.

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    Low-Temperature Biosurfactants from Polar Microbes
    (MDPI AG, 2020-08) Trudgeon, Benjamin; Dieser, Markus; Balasubramanian, Narayanaganesh; Messmer, Mitch; Foreman, Christine M.
    Surfactants, both synthetic and natural, are used in a wide range of industrial applications, including the degradation of petroleum hydrocarbons. Organisms from extreme environments are well-adapted to the harsh conditions and represent an exciting avenue of discovery of naturally occurring biosurfactants, yet microorganisms fromcold environments have been largely overlooked for their biotechnological potential as biosurfactant producers. In this study, four cold-adapted bacterial isolates from Antarctica are investigated for their ability to produce biosurfactants. Here we report on the physical properties and chemical structure of biosurfactants from the genera Janthinobacterium, Psychrobacter, and Serratia. These organisms were able to grow on diesel, motor oil, and crude oil at 4  C. Putative identification showed the presence of sophorolipids and rhamnolipids. Emulsion index test (E24) activity ranged from 36.4–66.7%. Oil displacement tests were comparable to 0.1–1.0% sodium dodecyl sulfate (SDS) solutions. Data presented herein are the first report of organisms of the genus Janthinobacterium to produce biosurfactants and their metabolic capabilities to degrade diverse petroleum hydrocarbons. The organisms’ ability to produce biosurfactants and grow on different hydrocarbons as their sole carbon and energy source at low temperatures (4  C) makes them suitable candidates for the exploration of hydrocarbon bioremediation in low-temperature environments.
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    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.
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    Quorum sensing inhibition as a promising method to control biofilm growth in metalworking fluids
    (2019-04) Ozcan, Safiye S.; Dieser, Markus; Parker, Albert E.; Balasubramanian, Narayanaganesh; Foreman, Christine M.
    Microbial contamination in metalworking systems is a critical problem. This study determined the microbial communities in metalworking fluids (MWFs) from two machining shops and investigated the effect of quorum sensing inhibition (QSI) on biofilm growth. In both operations, biofilm-associated and planktonic microbial communities were dominated by Pseudomonadales (60.2–99.7%). Rapid recolonization was observed even after dumping spent MWFs and meticulous cleaning. Using Pseudomonas aeruginosa PAO1 as a model biofilm organism, patulin (40 µM) and furanone C-30 (75 µM) were identified as effective QSI agents. Both agents had a substantially higher efficacy compared to α-amylase (extracellular polymeric substance degrading enzyme) and reduced biofilm formation by 63% and 76%, respectively, in MWF when compared to untreated controls. Reduced production of putatively identified homoserine lactones and quinoline in MWF treated with QS inhibitors support the effect of QSI on biofilm formation. The results highlight the effectiveness of QSI as a potential strategy to eradicate biofilms in MWFs.
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    DOM composition alters ecosystem function during microbial processing of isolated sources
    (2019-01) D'Andrilli, Juliana; Junker, James R.; Smith, Heidi J.; Scholl, Eric A.; Foreman, Christine M.
    Dynamics of dissolved organic matter (DOM) in ecosystems are controlled by a suite of interacting physical, chemical, and biological factors. Growing recognition of the associations between microbial communities and metabolism and intrinsic DOM characteristics, highlight the potential importance of microbe-DOM relationships to modulate the role and fate of DOM, yet these relationships are difficult to isolate because they often operate across confounding environmental gradients. In a controlled laboratory incubation (44 days), we integrated DOM bulk and molecular characterization, bacterial abundances, microbial assemblage composition, nutrient concentrations, and cellular respiration to discern the structural dynamics of biological processing among DOM sources from different allochthonous litters (grass, deciduous leaves, and evergreen needles). We identified two periods, consistent among DOM sources, where processing dynamics differed. Further, bulk fluorescent analyses showed shifts from low to high excitation and emission wavelengths, indicating the biological production of more complex/degraded materials over time. Molecular level analyses revealed similar temporal patterns among DOM sources in the production and consumption of individual chemical components varying in reactivity and heteroatomic content. Despite these similarities, total carbon (C) removed and carbon dioxide (CO2) accumulation differed by ~ 20% and 25% among DOM sources. This range in C processing was apparently tied to key chemical properties of the DOM (e.g., initial DOM composition, N content, and labile nature) as well as differential reorganization of the microbial populations that decomposed the DOM. We conclude that the production, transformation, and consumption of C in aquatic ecosystems is strongly dependent on the source and character of DOM as well as the structure of the microbial communities present, both of which change as DOM is processed over time. It is crucial that stream C processing models represent this complexity accurately.
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    Ultrafast Excited-State Deactivation of the Bacterial Pigment Violacein
    (2017-08) Beckstead, Ashley A.; Zhang, Yuyan; Hilmer, Jonathan K.; Smith, Heidi J.; Bermel, Emily; Foreman, Christine M.; Kohler, Bern
    The photophysical properties of the natural pigment violacein extracted from an Antarctic organism adapted to high exposure levels of UV radiation were measured in a combined steady-state and time-resolved spectroscopic study for the first time. In the low-viscosity solvents methanol and acetone, violacein exhibits low fluorescence quantum yields on the order of 1 x 10(-4), and femtosecond transient absorption measurements reveal excited-state lifetimes of 3.2 +/- 0.2 and 4.6 +/- 0.2 ps in methanol and acetone, respectively. As solvent viscosity is increased, both the fluorescence quantum yield and excited-state lifetime of this intensely colored pigment increase dramatically, and stimulated emission decays 30-fold more slowly in glycerol than in methanol at room temperature. Excited-state deactivation is suggested to occur via a molecular-rotor mechanism in which torsion interring bond leads to a conical intersection with the ground state.
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    A 21 000-year record of fluorescent organic matter markers in the WAIS Divide ice core
    (2017-05) D'Andrilli, Juliana; Foreman, Christine M.; Sigl, Michael; Priscu, John C.; McConnell, Joseph R.
    Englacial ice contains a significant reservoir of organic material (OM), preserving a chronological record of materials from Earth's past. Here, we investigate if OM composition surveys in ice core research can provide paleoecological information on the dynamic nature of our Earth through time. Temporal trends in OM composition from the early Holocene extending back to the Last Glacial Maximum (LGM) of the West Antarctic Ice Sheet Divide (WD) ice core were measured by fluorescence spectroscopy. Multivariate parallel factor (PARAFAC) analysis is widely used to isolate the chemical components that best describe the observed variation across three-dimensional fluorescence spectroscopy (excitation–emission matrices; EEMs) assays. Fluorescent OM markers identified by PARAFAC modeling of the EEMs from the LGM (27.0–18.0 kyr BP; before present 1950) through the last deglaciation (LD; 18.0–11.5 kyr BP), to the mid-Holocene (11.5–6.0 kyr BP) provided evidence of different types of fluorescent OM composition and origin in the WD ice core over 21.0 kyr. Low excitation–emission wavelength fluorescent PARAFAC component one (C1), associated with chemical species similar to simple lignin phenols was the greatest contributor throughout the ice core, suggesting a strong signature of terrestrial OM in all climate periods. The component two (C2) OM marker, encompassed distinct variability in the ice core describing chemical species similar to tannin- and phenylalanine-like material. Component three (C3), associated with humic-like terrestrial material further resistant to biodegradation, was only characteristic of the Holocene, suggesting that more complex organic polymers such as lignins or tannins may be an ecological marker of warmer climates. We suggest that fluorescent OM markers observed during the LGM were the result of greater continental dust loading of lignin precursor (monolignol) material in a drier climate, with lower marine influences when sea ice extent was higher and continents had more expansive tundra cover. As the climate warmed, the record of OM markers in the WD ice core changed, reflecting shifts in carbon productivity as a result of global ecosystem response..
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    Biogeophysical properties of an expansive Antarctic supraglacial stream
    (2017) SanClements, M. D.; Michael, D.; Smith, Heidi J.; Foreman, Christine M.; Tedesco, Marco; Chin, Yu-Ping
    Supraglacial streams are important hydrologic features in glaciated environments as they are conduits for the transport of aeolian debris, meltwater, solutes and microbial communities. We characterized the basic geomorphology, hydrology and biogeochemistry of the Cotton Glacier supraglacial stream located in the McMurdo Dry Valleys of Antarctica. The distinctive geomorphology of the stream is driven by accumulated aeolian sediment from the Transantarctic Mountains, while solar radiation and summer temperatures govern melt in the system. The hydrologic functioning of the Cotton Glacier stream is largely controlled by the formation of ice dams that lead to vastly different annual flow regimes and extreme flushing events. Stream water is chemically dilute and lacks a detectable humic signature. However, the fluorescent signature of dissolved organic matter (DOM) in the stream does demonstrate an extremely transitory red-shifted signal found only in near-stream sediment leachates and during the initial flushing of the system at the onset of flow. This suggests that episodic physical flushing drives pulses of DOM with variable quality in this stream. This is the first description of a large Antarctic supraglacial stream and our results provide evidence that the hydrology and geomorphology of supraglacial streams drive resident microbial community composition and biogeochemical cycling
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    Geomicrobiology of Blood Falls: An iron-rich saline discharge at terminus of the Taylor Glacier, Antarctica
    (2004-09) Mikucki, Jill A.; Foreman, Christine M.; Sattler, Birgit; Lyons, W. Berry; Priscu, John C.
    Blood Falls, a saline subglacial discharge from the Taylor Glacier, Antarctica provides an example of the diverse physical and chemical niches available for life in the polar desert of the McMurdo Dry Valleys. Geochemical analysis of Blood Falls outflow resembles concentrated seawater remnant from the Pliocene intrusion of marine waters combined with products of weathering. The result is an iron-rich, salty seep at the terminus of Taylor Glacier, which is subject to episodic releases into permanently ice-covered Lake Bonney. Blood Falls influences the geochemistry of Lake Bonney, and provides organic carbon and viable microbes to the lakesystem. Here we present the first data on the geobiology of Blood Falls and relate it to the evolutionary history of this unique environment. The novel geological evolution of this subglacial environment makes Blood Falls an important site for the study of metabolic strategies in subglacial environments and the impact of subglacial efflux on associated lake ecosystems.
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    Impact of episodic warming events
    (2004-09) Foreman, Christine M.; Wolf, Craig F.; Priscu, John C.
    Lakes in the Taylor Valley, Antarctica, were investigated to determine the impact of a significant air temperature warming event that occurred during the austral summer of 2001–2002. The warming in the valleys caused an increase in glacial run-off, record stream discharge, an increase in lake levels, and thinning of the permanent ice covers. These changes in the physical environment drove subsequent changes in the biogeochemistry of the lakes. Primary production in West Lake Bonney during the flood was reduced 23% as a consequence of stream induced water column turbidity. Increased nutrient levels within the lakes occurred in the year following the temperature induced high flow year. For example, soluble reactive phosphorus loading to Lake Fryxell was four-fold greater than the long-term average loading rates. These high nutrient levels corresponded to an increase in primary production in the upper water columns of Lakes Bonney and Fryxell. Depth integrated chlorophyll-a values increased 149% in East Lake Bonney, 48% in West Lake Bonney, and showed little change in Lake Fryxell; chlorophyll-a in Lake Hoare decreased 18% compared to long-term averages recorded as part of our ten year monitoring program, presumably from a reduction in under-ice PAR caused by increased sediment loads on the ice cover. Overall the warming event served to recharge the ecosystem with liquid water and associated nutrients. Such floods may play an important role in the long-term maintenance of liquid water in these dry valley lakes.
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    Glacial ice cores: A model system for developing extraterrestrial decontamination protocols
    (2005-04) Christner, Brent C.; Mikucki, Jill A.; Foreman, Christine M.; Denson, Jackie; Priscu, John C.
    Evidence gathered from spacecraft orbiting Mars has shown that water ice exists at both poles and may form a large subsurface reservoir at lower latitudes. The recent exploration of the martian surface by unmanned landers and surface rovers, and the planned missions to eventually return samples to Earth have raised concerns regarding both forward and back contamination. Methods to search for life in these icy environments and adequate protocols to prevent contamination can be tested with earthly analogues. Studies of ice cores on Earth have established past climate changes and geological events, both globally and regionally, but only recently have these results been correlated with the biological materials (i.e., plant fragments, seeds, pollen grains, fungal spores, and microorganisms) that are entrapped and preserved within the ice. The inclusion of biology into ice coring research brings with it a whole new approach towards decontamination. Our investigations on ice from the Vostok core (Antarctica) have shown that the outer portion of the cores have up to 3 and 2 orders of magnitude higher bacterial density and dissolved organic carbon (DOC) than the inner portion of the cores, respectively, as a result of drilling and handling. The extreme gradients that exist between the outer and inner portion of these samples make contamination a very relevant aspect of geomicrobiological investigations with ice cores, particularly when the actual numbers of ambient bacterial cells are low. To address this issue and the inherent concern it raises for the integrity of future investigations with ice core materials from terrestrial and extraterrestrial environments, we employed a procedure to monitor the decontamination process in which ice core surfaces are painted with a solution containing a tracer microorganism, plasmid DNA, and fluorescent dye before sampling. Using this approach, a simple and direct method is proposed to verify the authenticity of geomicrobiological results obtained from ice core materials. Our protocol has important implications for the design of life detection experiments on Mars and the decontamination of samples that will eventually be returned to Earth.
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