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            Biddle, Jennifer F (Ed.)ABSTRACT Autotrophic bacteria are able to fix CO2in a great diversity of habitats, even though this dissolved gas is relatively scarce at neutral pH and above. As many of these bacteria rely on CO2fixation by ribulose 1,5-bisphospate carboxylase/oxygenase (RubisCO) for biomass generation, they must compensate for the catalytical constraints of this enzyme with CO2-concentrating mechanisms (CCMs). CCMs consist of CO2and HCO3−transporters and carboxysomes. Carboxysomes encapsulate RubisCO and carbonic anhydrase (CA) within a protein shell and are essential for the operation of a CCM in autotrophicBacteriathat use the Calvin-Benson-Basham cycle. Members of the genusThiomicrospiralack genes homologous to those encoding previously described CA, and prior to this work, the mechanism of function for their carboxysomes was unclear. In this paper, we provide evidence that a member of the recently discovered iota family of carbonic anhydrase enzymes (ιCA) plays a role in CO2fixation by carboxysomes from members ofThiomicrospiraand potentially otherBacteria. Carboxysome enrichments fromThiomicrospira pelophilaandThiomicrospira aerophilawere found to have CA activity and contain ιCA, which is encoded in their carboxysome loci. When the gene encoding ιCA was interrupted inT. pelophila, cells could no longer grow under low-CO2conditions, and CA activity was no longer detectable in their carboxysomes. WhenT. pelophilaιCA was expressed in a strain ofEscherichia colilacking native CA activity, this strain recovered an ability to grow under low CO2conditions, and CA activity was present in crude cell extracts prepared from this strain. IMPORTANCEHere, we provide evidence that iota carbonic anhydrase (ιCA) plays a role in CO2fixation by some organisms with CO2-concentrating mechanisms; this is the first time that ιCA has been detected in carboxysomes. While ιCA genes have been previously described in other members of bacteria, this is the first description of a physiological role for this type of carbonic anhydrase in this domain. Given its distribution in alkaliphilic autotrophic bacteria, ιCA may provide an advantage to organisms growing at high pH values and could be helpful for engineering autotrophic organisms to synthesize compounds of industrial interest under alkaline conditions.more » « less
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            Abstract Coastal wetlands such as mangrove forests are breeding grounds for nuisance-causing insects. Rotational Impoundment Management (RIM) for mosquito control involves annual summer inundation of impounded mangrove forests with estuarine water during the summer half year. However, in addition to controlling mosquitos, RIM may change biogeochemical pathways. This study set out to investigate how RIM quantitatively affects physicochemical soil characteristics and potential nitrifying and denitrifying activities (PNA and PDA), which are key in the global nitrogen cycle. Before and after the implementation of RIM, soil samples were collected annually in habitats differing in size and abundance of black mangroves (Avicennia germinans) in an impoundment with RIM and in an adjacent impoundment with a more open connection to the lagoon. Compared to the non-managed impoundment, soil moisture content, total nitrogen and PDA increased, while salinity decreased after the start of annual summer flooding, but only in the dwarf habitat. In the sparse and dense habitats, total nitrogen and PDA increased independently of summer flooding, whereas soil moisture content and salinity were not affected by RIM. Labile organic nitrogen increased only in the RIM impoundment, irrespective of the habitat type. PNA was generally not affected with time, except in the dwarf habitat in the absence of intentional summer flooding where it increased. Changes in the non-managed impoundment adjacent to the RIM impoundment demonstrate the importance of groundwater exchange in linked ecosystems. The consequences of interventions in the management of mangrove impoundments and adjacent forests for the nitrogen budget are discussed.more » « lessFree, publicly-accessible full text available December 1, 2025
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            Abstract The extended phenotype of helical burrowing behavior in animals has evolved independently many times since the Cambrian explosion (~540 million years ago [MYA]). A number of hypotheses have been proposed to explain the evolution of helical burrowing in certain taxa, but no study has searched for a general explanation encompassing all taxa. We reviewed helical burrowing in both extant and extinct animals and from the trace fossil record and compiled 10 hypotheses for why animals construct helical burrows, including our own ideas. Of these, six are post‐construction hypotheses—benefits to the creator or offspring, realized after burrow construction—and four are construction hypotheses reflecting direct benefits to the creator during construction. We examine the fit of these hypotheses to a total of 21 extant taxa and ichnotaxa representing 59–184 possible species. Only two hypotheses, antipredator and biomechanical advantage, cannot be rejected for any species (possible in 100% of taxa), but six of the hypotheses cannot be rejected for most species (possible in 86%–100% of taxa): microclimate buffer, reduced falling sediment (soil), anticrowding, and vertical patch. Four of these six are construction hypotheses, raising the possibility that helical burrowing may have evolved without providing post‐construction benefits. Our analysis shows that increased drainage, deposit feeding, microbial farming, and offspring escape cannot explain helical burrowing behavior in the majority of taxa (5%–48%). Overall, the evidence does not support a general explanation for the evolution of helical burrowing in animals. The function and evolution of the helix as an extended phenotype seems to provide different advantages for different taxa in different environments under different physicochemical controls (some traces/tracemakers are discussed in more detail due to their association with body fossils and well‐constrained physicochemical parameters). Although direct tests of many of the hypotheses would be difficult, we nevertheless offer ways to test some of the hypotheses for selected taxa.more » « less
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            Abstract Between the 1780 and 1980s, more than half of the wetlands in the conterminous US were lost. As wetlands have been lost, numerous artificial water features (AWFs), such as stormwater retention ponds, golf course water features, and reservoirs, have been constructed. We contrasted the loss of wetland area and perimeter to the gain of AWF area and perimeter and further explored how this transformation has altered the spatial characteristics of the waterscape. We conducted this analysis in the Tampa Bay Watershed, a large coastal watershed that lost 33% of its wetland area between the 1950s-2007. Trends have been towards fewer, smaller wetlands and more, smaller AWFs. The loss of wetland area far exceeds the gain in AWF area, leading to an overall loss of 23% of the combined wetland and AWF area. However, the loss of wetland perimeter almost equals the gain in AWF perimeter, leading to an overall loss of just 2% of the combined wetland and AWF perimeter. The loss of wetlands and gain of AWFs have predominantly occurred in different geographic locations, with the loss of wetlands predominantly in the headwaters and the gain in AWFs predominantly adjacent to Tampa Bay. Wetlands became further apart, though generally retained their natural distribution, while AWFs became closer to one another and now mirror the more natural wetland distribution. Overall, the physical structure of the waterscape of today is different than in the past, which likely reflects a change in functions performed and related ecological services provided at local and landscape scales.more » « less
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            Abstract Many headwater wetlands are integrated into flowpath networks and can serve as sources of streamflow for downgradient waters. We demonstrate this with five years of data in vernal pool, swale, and headwater stream complexes in the Central Valley, California. Long-term United States Geological Survey data suggest that the mean flow duration from the smallest watersheds in this region, including those with vernal pool, swale, and headwater stream complexes, is ~ 85 days per year. Our data concur, indicating that the annual days of flow per year from our vernal pool, swale, and headwater stream complexes ranges from ~ 20–200, but is ~ 85 when annual precipitation is 100% of normal. Peak stages are evident first in vernal pools which then propagate sequentially downstream through swales, headwater streams, and to the Sacramento River at celerities of ~ 1-1.5 m/s, consistent with expected flood wave velocities. Geospatial analyses show that these vernal pool, swale, and headwater stream features cover > 4% of the study area. Our results suggest these systems can be significant sources of streamflow, and therefore play an important role in maintaining the chemical, physical, and biological integrity of downstream waters, which has important implications for the definition of waters of the United States subject to regulation under the Clean Water Act.more » « less
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            Abstract Watershed resilience is the ability of a watershed to maintain its characteristic system state while concurrently resisting, adapting to, and reorganizing after hydrological (for example, drought, flooding) or biogeochemical (for example, excessive nutrient) disturbances. Vulnerable waters include non-floodplain wetlands and headwater streams, abundant watershed components representing the most distal extent of the freshwater aquatic network. Vulnerable waters are hydrologically dynamic and biogeochemically reactive aquatic systems, storing, processing, and releasing water and entrained (that is, dissolved and particulate) materials along expanding and contracting aquatic networks. The hydrological and biogeochemical functions emerging from these processes affect the magnitude, frequency, timing, duration, storage, and rate of change of material and energy fluxes among watershed components and to downstream waters, thereby maintaining watershed states and imparting watershed resilience. We present here a conceptual framework for understanding how vulnerable waters confer watershed resilience. We demonstrate how individual and cumulative vulnerable-water modifications (for example, reduced extent, altered connectivity) affect watershed-scale hydrological and biogeochemical disturbance response and recovery, which decreases watershed resilience and can trigger transitions across thresholds to alternative watershed states (for example, states conducive to increased flood frequency or nutrient concentrations). We subsequently describe how resilient watersheds require spatial heterogeneity and temporal variability in hydrological and biogeochemical interactions between terrestrial systems and down-gradient waters, which necessitates attention to the conservation and restoration of vulnerable waters and their downstream connectivity gradients. To conclude, we provide actionable principles for resilient watersheds and articulate research needs to further watershed resilience science and vulnerable-water management.more » « less
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            Abstract Stream dissolved oxygen (DO) dynamics are an outcome of metabolic activity and subsequently regulate ecosystem functions such as in‐stream solute and sediment reactions. The synchronization of DO signals in and across stream networks is both a cause and effect of the mode and timing of these functions, but there is limited empirical evidence for network patterns of DO synchrony. We used high frequency DO measurements at 42 sites spanning five catchments and stream orders to evaluate DO signal synchrony in response to variation in light (a driver of photosynthesis) and discharge (a control on DO signal spatial extent). We hypothesized that stream network DO synchrony arises when regional controls dominate: when light inputs are synchronous and when longitudinal hydrologic connectivity is high. By complement, we predicted that DO signal synchrony decreases as light becomes more asynchronous and stream flows decline or become discontinuous. Our results supported this hypothesis: greater DO signal synchrony arose with increasing light synchrony and flow connectivity. A model including these two controls explained 70% of variation in DO synchrony. We conclude that DO synchrony patterns within‐ and across‐networks support the current paradigm of discharge and light control on stream metabolic activity. Finally, we propose that DO synchrony patterns are likely a useful prerequisite for scaling subdaily metabolism estimates to network and regional scales.more » « less
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            Free, publicly-accessible full text available December 1, 2025
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            Groundwater extraction compromises the function of groundwater-dependent ecosystems, such as freshwater wetlands. Identifying whether groundwater conservation restores wetland hydrology is a first step toward rehabilitating impaired wetlands. In the Tampa Bay region of Florida (U.S.), groundwater extraction rates have been declining since 1998, partly in response to desiccation of wetlands and waterbodies. This study uses monthly water-level data from 152 depressional wetlands over 28 years (1991–2018) to identify trends in wetland inundation, determine whether those trends vary among wetlands historically exposed to different rates of groundwater extraction, and describe relationships between the timing and extent of cutbacks in groundwater extraction and the timing and extent of changes in wetland inundation. Many wetlands (57 %) exhibited increased inundation in response to cutbacks in groundwater extraction, indicating that water conservation measures are inducing recovery. Further, increased inundation began in most wetlands immediately upon, or within two years of, the time extraction cutbacks occurred, although some recovering wetlands exhibited longer lags. An additional 26 % of wetlands had steady-state water levels with inundation similar to that of reference wetlands, potentially revealing a population of wetlands hydrologically unimpaired by nearby groundwater extraction. Another subset of wetlands (14 %) with steady-state water depths exhibited increasing deviations from basin-full water levels, suggesting subsidence of the wetland basin. Active intervention beyond cutbacks in groundwater extraction may be necessary to restore this subset, whereas passive restoration (reducing extraction) appears adequate for the majority of impacted wetlands. Rising water levels may amplify surface-water connections among wetlands, with ecological and biogeochemical consequences both for individual wetlands and for the whole wetlandscape. As a host of human activities continue to rely on groundwater extraction, this study demonstrates the potential for, as well as variability in, hydrological recovery across a wetland-rich, low-relief landscape following the enactment of water conservation policies.more » « lessFree, publicly-accessible full text available November 1, 2025
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            In communities where people lack on-demand, safely managed drinking water, stored household water often becomes contaminated by fecal bacteria, regardless of the source-water quality. The objectives of this paper are to assess and control bacterial contamination in stored household water in Toamasina, a rapidly urbanizing city in eastern coastal Madagascar. We collected samples of source water and stored household water from 10 representative households that use different water sources and different storage strategies, and we analyzed the samples for several fecal indicator bacteria. We also tested three methods that residents of Toamasina could realistically employ for cleaning their household water storage vessels, assessing the effect of the cleaning methods on measured bacterial levels in the water. Consistent with the previous literature, we found that concentrations of total coliforms in stored household water were significantly higher than in the corresponding source water (p < 0.05). In 100% of households that stored their water in 20 L polyethylene jerrycans (n = 4), biofilms on the walls of the jerrycan harbored total coliforms and Enterococcus. The use of a closed storage container was, on its own, not found to provide a meaningful protective effect against bacterial regrowth; to be protective, closed storage containers must be combined with high-quality source water and/or with adequate cleaning to prevent biofilm formation. A dilute solution of sodium hypochlorite, known locally as Sûr’Eau or Manadio Rano, was both the most effective and the least expensive method for cleaning household water storage containers. We conclude that regular and effective cleaning of storage containers is an essential component of safe water storage. Because household storage of collected water is common in many low- and middle-income countries, these results are important towards the worldwide achievement of the United Nations’ Sustainable Development Goal 6.more » « less
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