Search

ABSTRACT Twitching motility is a form of bacterial surface translocation powered by the type IV pilus (T4P). It is frequently analyzed by interstitial colony expansion between agar and the polystyrene surfaces of petri dishes. In such assays, the twitching motility of Acinetobacter nosocomialis was observed with MacConkey but not Luria-Bertani (LB) agar media. One difference between these two media is the presence of bile salts as a selective agent in MacConkey but not in LB. Here, we demonstrate that the addition of bile salts to LB allowed A. nosocomialis to display twitching. Similarly, bile salts enhanced the twitching of Acinetobacter baumannii and Pseudomonas aeruginosa in LB. These observations suggest that there is a common mechanism, whereby bile salts enhance bacterial twitching and promote interstitial colony expansion. Bile salts disrupt lipid membranes and apply envelope stress as detergents. Surprisingly, their stimulatory effect on twitching appears not to be related to a bacterial physiological response to stressors. Rather, it is due to their ability to alter the physicochemical properties of a twitching surface. We observed that while other detergents promoted twitching like bile salts, stresses applied by antibiotics, including the outer membrane-targeting polymyxin B, did not enhance twitching motility. More importantly, bacteria displayed increased twitching on hydrophilic surfaces such as those of glass and tissue culture-treated polystyrene plastics, and bile salts no longer stimulated twitching on these surfaces. Together, our results show that altering the hydrophilicity of a twitching surface significantly impacts T4P functionality.IMPORTANCEThe bacterial type IV pilus (T4P) is a critical virulence factor for many medically important pathogens, some of which are prioritized by the World Health Organization for their high levels of antibiotic resistance. The T4P is known to propel bacterial twitching motility, the analysis of which provides a convenient assay for T4P functionality. Here, we show that bile salts and other detergents augment the twitching of multiple bacterial pathogens. We identified the underlying mechanism as the alteration of surface hydrophilicity by detergents. Consequently, hydrophilic surfaces like those of glass or plasma-treated polystyrene promote bacterial twitching, bypassing the requirement for detergents. The implication is that surface properties, such as those of tissues and medical implants, significantly impact the functionality of bacterial T4P as a virulence determinant. This offers valuable insights for developing countermeasures against the colonization and infection by bacterial pathogens of critical importance to human health on a global scale.

Abstract Several moons in the outer solar system host liquid water oceans. A key next step in assessing the habitability of these ocean worlds is to determine whether life’s elemental and energy requirements are also met. Phosphorus is required by all known life and is often limited to biological productivity in Earth’s oceans. This raises the possibility that its availability may limit the abundance or productivity of Earth-like life on ocean worlds. To address this potential problem, here we calculate the equilibrium dissolved phosphate concentrations associated with the reaction of water and rocks—a key driver of ocean chemical evolution—across a broad range of compositional inputs and reaction conditions. Equilibrium dissolved phosphate concentrations range from 10−11 to 10−1 mol/kg across the full range of carbonaceous chondrite compositions and reaction conditions considered, but are generally > 10−5 mol/kg for most plausible scenarios. Relative to the phosphate requirements and uptake kinetics of microorganisms in Earth’s oceans, such concentrations would be sufficient to support initially rapid cell growth and construction of global ocean cell populations larger than those observed in Earth’s deep oceans.

Cryopeg brines are isolated volumes of hypersaline water in subzero permafrost. The cryopeg system at Utqiaġvik, Alaska, is estimated to date back to 40 ka BP or earlier, a remnant of a late Pleistocene Ocean. Surprisingly, the cryopeg brines contain high concentrations of organic carbon, including extracellular polysaccharides, and high densities of bacteria. How can these physiologically extreme, old, and geologically isolated systems support such an ecosystem? This study addresses this question by examining the energetics of the Utqiaġvik cryopeg brine ecosystem. Using literature-derived assumptions and new measurements on archived borehole materials, we first estimated the quantity of organic carbon when the system formed. We then considered two bacterial growth trajectories to calculate the lower and upper bounds of the cell-specific metabolic rate of these communities. These bounds represent the first community estimates of metabolic rate in a subzero hypersaline environment. To assess the plausibility of the different growth trajectories, we developed a model of the organic carbon cycle and applied it to three borehole scenarios. We also used dissolved inorganic carbon and nitrogen measurements to independently estimate the metabolic rate. The model reconstructs the growth trajectory of the microbial community and predicts the present-day cell density and organic carbon content. Model input included measured rates of the in-situ enzymatic conversion of particulate to dissolved organic carbon under subzero brine conditions. A sensitivity analysis of model parameters was performed, revealing an interplay between growth rate, cell-specific metabolic rate, and extracellular enzyme activity. This approach allowed us to identify plausible growth trajectories consistent with the observed bacterial densities in the cryopeg brines. We found that the cell-specific metabolic rate in this system is relatively high compared to marine sediments. We attribute this finding to the need to invest energy in the production of extracellular enzymes, for generating bioavailable carbon from particulate organic carbon, and the production of extracellular polysaccharides for cryoprotection and osmoprotection. These results may be relevant to other isolated systems in the polar regions of Earth and to possible ice-bound brines on worlds such as Europa, Enceladus, and Mars.

ABSTRACT In recent years, our appreciation of the extent of habitable environments in Earth’s subsurface has greatly expanded, as has our understanding of the biodiversity contained within. Most studies have relied on single sampling points, rather than considering the long-term dynamics of subsurface environments and their microbial populations. One such habitat are aquifers associated with the aqueous alteration of ultramafic rocks through a process known as serpentinization. Ecological modeling performed on a multiyear time series of microbiology, hydrology, and geochemistry in an ultrabasic aquifer within the Coast Range Ophiolite reveals that community assembly is governed by undominated assembly (i.e., neither stochastic [random] nor deterministic [selective] processes alone govern assembly). Controls on community assembly were further assessed by characterizing aquifer hydrogeology and microbial community adaptations to the environment. These analyses show that low permeability rocks in the aquifer restrict the transmission of microbial populations between closely situated wells. Alpha and beta diversity measures and metagenomic and metatranscriptomic data from microbial communities indicate that high pH and low dissolved inorganic carbon levels impose strong environmental selection on microbial communities within individual wells. Here, we find that the interaction between strong selection imposed by extreme pH and enhanced ecological drift due to dispersal limitation imposed by slow fluid flow results in the undominated assembly signal observed throughout the site. Strong environmental selection paired with extremely low dispersal in the subsurface results in low diversity microbial communities that are well adapted to extreme pH conditions and subject to enhanced stochasticity introduced by ecological drift over time. IMPORTANCE Microbial communities existing under extreme or stressful conditions have long been thought to be structured primarily by deterministic processes. The application of macroecology theory and modeling to microbial communities in recent years has spurred assessment of assembly processes in microbial communities, revealing that both stochastic and deterministic processes are at play to different extents within natural environments. We show that low diversity microbial communities in a hard-rock serpentinizing aquifer are assembled under the influence of strong selective processes imposed by high pH and enhanced ecological drift that occurs as the result of dispersal limitation due to the slow movement of water in the low permeability aquifer. This study demonstrates the important roles that both selection and dispersal limitation play in terrestrial serpentinites, where extreme pH assembles a microbial metacommunity well adapted to alkaline conditions and dispersal limitation drives compositional differences in microbial community composition between local communities in the subsurface.

In the search for life on other planets, the presence of photosynthetic surface vegetation may be detectable from the colors of light it reflects. On the modern Earth, this spectral reflectance is characterized by a steep increase in reflectance between the red and near‐infrared wavelengths, a signature known as the “red edge”. This edge-like signature occurs at wavelengths of peak photon absorbance, which are the result of adaptations of the phototroph to their spectral environment. On planets orbiting different stellar types, red edge analogs may occur at other colors than red. Thus, knowing the wavelengths at which photosynthetic organisms preferentially absorb and reflect photons is necessary to detect red edge analogs on other planets. Using a numerical model that predicts the absorbance spectrum of extant photosynthetic pigments on Earth from Marosvölgyi and van Gorkom (2010), we calculate the absorbance spectrum for pigments on an Earth-like planet around F through late M type stars that are adapted for maximal energy production. In this model, cellular energy production is maximized when pigments are tuned to absorb at the wavelength that maximizes energy input from incident photons while minimizing energy losses due to thermal emission and building cellular photosynthetic apparatus. We find that peak photon absorption for photosynthetic organisms around F type stars tends to be in the blue while for G, K, and early M type stars, red or just beyond is preferred. Around the coolest M type stars, these organisms may preferentially absorb in the near-infrared, possibly past one micron. These predictions are consistent with previous, qualitative estimates of pigment absorptance. Our predicted absorbance spectra for photosynthetic surface organisms depend on both the stellar type and planetary atmospheric composition, especially atmospheric water vapor concentrations, which alter the availability of surface photons and thus the predicted pigment absorption. By constraining the absorbance spectra of alien, photosynthetic organisms, future observations may be better equipped to detect the weak spectral signal of red edge analogs.

Bloom's taxonomy is a classification of learning objectives originally developed for general educational purposes. The taxonomy was revised to expand beyond cognitive processes and to include knowledge types as an orthogonal dimension. As Bloom's taxonomy is a tool widely used in biology education by researchers and instructors, it is important to examine the underlying assumptions embedded within how people may implicitly understand and use the taxonomy. In this paper, we empirically examine two major assumptions: the independence of the knowledge-type and cognitive-process dimensions and the use of action verbs as proxies for different cognitive processes. Contingency analysis on 940 assessment items revealed that the knowledge-type and cognitive-process dimensions are related and not independent. Subsequent correspondence analysis identified two principle axes in how the two dimensions are related, with three clusters of knowledge types and cognitive processes. Using the Shannon evenness index, we did not find a clear relationship between question prompt words (including action verbs) and cognitive processes in the assessment items. Based on these results, we suggest that both dimensions of the revised Bloom's taxonomy should be used and that question prompt words or action verbs alone are not sufficient in classifying the embedded learning objectives within assessment items.

Past studies of hydrogen cycling in hypersaline microbial mats have shown an active nighttime cycle, with production largely from Cyanobacteria and consumption from sulfate-reducing bacteria (SRB). However, the mechanisms and magnitude of hydrogen cycling have not been extensively studied. Two mats types near Guerrero Negro, Mexico -- permanently submerged Microcoleus microbial mats (GN-S), and intertidal Lyngbya microbial mats (GN-I) -- were used in microcosm diel manipulation experiments with 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU), molybdate, ammonium addition, and physical disruption to understand the processes responsible for hydrogen cycling between mat microbes. Across microcosms, H2 production occurred under dark anoxic conditions with simultaneous production of a suite of organic acids. H2 production was not significantly affected by inhibition of nitrogen fixation, but rather appears to result from constitutive fermentation of photosynthetic storage products by oxygenic phototrophs. Comparison to accumulated glycogen and to CO2 flux indicated that, in the GN-I mat, fermentation released almost all of the carbon fixed via photosynthesis during the preceding day, primarily as organic acids. Across mats, although oxygenic and anoxygenic phototrophs were detected, cyanobacterial [NiFe]-hydrogenase transcripts predominated. Molybdate inhibition experiments indicated that SRBs from a wide distribution of dsrA phylotypes were responsible for H2 consumption. Incubation with 13C-acetate and nanoSIMS (secondary ion mass-spectrometry) indicated higher uptake in both Chloroflexi and SRBs relative to other filamentous bacteria. These manipulations and diel incubations confirm that Cyanobacteria were the main fermenters in Guerrero Negro mats and that the net flux of nighttime fermentation byproducts (not only hydrogen) was largely regulated by the interplay between Cyanobacteria, SRBs, and Chloroflexi.

Bloom's taxonomy is a classification of learning objectives originally developed for general educational purposes. The taxonomy was revised to expand beyond cognitive processes and to include knowledge types as an orthogonal dimension. As Bloom's taxonomy is a tool widely used in biology education by researchers and instructors, it is important to examine the underlying assumptions embedded within how people may implicitly understand and use the taxonomy. In this paper, we empirically examine two major assumptions: the independence of the knowledge-type and cognitive-process dimensions and the use of action verbs as proxies for different cognitive processes. Contingency analysis on 940 assessment items revealed that the knowledge-type and cognitive-process dimensions are related and not independent. Subsequent correspondence analysis identified two principle axes in how the two dimensions are related, with three clusters of knowledge types and cognitive processes. Using the Shannon evenness index, we did not find a clear relationship between question prompt words (including action verbs) and cognitive processes in the assessment items. Based on these results, we suggest that both dimensions of the revised Bloom's taxonomy should be used and that question prompt words or action verbs alone are not sufficient in classifying the embedded learning objectives within assessment items.

Cryopeg brines are isolated volumes of hypersaline water in subzero permafrost. The cryopeg system at Utqiaġvik, Alaska, is estimated to date back to 40 ka BP or earlier, a remnant of a late Pleistocene Ocean. Surprisingly, the cryopeg brines contain high concentrations of organic carbon, including extracellular polysaccharides, and high densities of bacteria. How can these physiologically extreme, old, and geologically isolated systems support such an ecosystem? This study addresses this question by examining the energetics of the Utqiaġvik cryopeg brine ecosystem. Using literature-derived assumptions and new measurements on archived borehole materials, we first estimated the quantity of organic carbon when the system formed. We then considered two bacterial growth trajectories to calculate the lower and upper bounds of the cellspecific metabolic rate of these communities. These bounds represent the first community estimates of metabolic rate in a subzero hypersaline environment. To assess the plausibility of the different growth trajectories, we developed a model of the organic carbon cycle and applied it to three borehole scenarios. We also used dissolved inorganic carbon and nitrogen measurements to independently estimate the metabolic rate. The model reconstructs the growth trajectory of the microbial community and predicts the present-day cell density and organic carbon content. Model input included measured rates of the in-situ enzymatic conversion of particulate to dissolved organic carbon under subzero brine conditions. A sensitivity analysis of model parameters was performed, revealing an interplay between growth rate, cell-specific metabolic rate, and extracellular enzyme activity. This approach allowed us to identify plausible growth trajectories consistent with the observed bacterial densities in the cryopeg brines. We found that the cell-specific metabolic rate in this system is relatively high compared to marine sediments. We attribute this finding to the need to invest energy in the production of extracellular enzymes, for generating bioavailable carbon from particulate organic carbon, and the production of extracellular polysaccharides for cryoprotection and osmoprotection. These results may be relevant to other isolated systems in the polar regions of Earth and to possible ice-bound brines on worlds such as Europa, Enceladus, and Mars. [ABSTRACT FROM AUTHOR]

There is unambiguous evidence that glaciers have retreated from their 19th century positions, but it is less clear how far glaciers have retreated relative to their long-term Holocene fluctuations. Glaciers in western North America are thought to have advanced from minimum positions in the Early Holocene to maximum positions in the Late Holocene. We assess when four North American glaciers, located between 38–60 ∘ N, were larger or smaller than their modern (2018–2020 CE) positions during the Holocene. We measured 26 paired cosmogenic in situ 14 C and 10 Be concentrations in recently exposed proglacial bedrock and applied a Monte Carlo forward model to reconstruct plausible bedrock exposure–burial histories. We find that these glaciers advanced past their modern positions thousands of years apart in the Holocene: a glacier in the Juneau Icefield (BC, Canada) at ∼2 ka, Kokanee Glacier (BC, Canada) at ∼6 ka, and Mammoth Glacier (WY, USA) at ∼1 ka; the fourth glacier, Conness Glacier (CA, USA), was likely larger than its modern position for the duration of the Holocene until present. The disparate Holocene exposure–burial histories are at odds with expectations of similar glacier histories given the presumed shared climate forcings of decreasing Northern Hemisphere summer insolation through the Holocene followed by global greenhouse gas forcing in the industrial era. We hypothesize that the range in histories is the result of unequal amounts of modern retreat relative to each glacier's Holocene maximum position, rather than asynchronous Holocene advance histories. We explore the influence of glacier hypsometry and response time on glacier retreat in the industrial era as a potential cause of the non-uniform burial durations. We also report mean abrasion rates at three of the four glaciers: Juneau Icefield Glacier (0.3±0.3 mm yr -1), Kokanee Glacier (0.04±0.03 mm yr -1), and Mammoth Glacier (0.2±0.2 mm yr -1). [ABSTRACT FROM AUTHOR]