Utility of far-field effects from tip-assisted Raman spectroscopy for the detection of a monolayer of diblock copolymer reverse micelles for nanolithography.

A modified set-up for Raman spectroscopy is proposed to utilize an AFM probe in a regime beyond the dependence on near field optics. Possible mechanisms for the observed enhancement have been explored through comparisons to spectra from other enhanced Raman techniques, including surface enhanced Raman, interference enhanced Raman and polarized Raman spectroscopies. The effects of polarization, focusing and interference are heightened when near field effects are diminished, giving rise to spectral enhancement. This technique allows for the characterization of a sub-20 nm monolayer of polystyrene-block-poly(2 vinyl pyridine) reverse micelles and paves the way for a promising method of non-destructive analysis of large self-assembled arrays of colloids.

Dynamic polyphosphate metabolism in cyanobacteria responding to phosphorus availability.

Despite the crucial role of polyphosphate (polyP) in aquatic environments, its metabolism in cyanobacteria responding to nutrients is poorly understood. We investigate polyP in three cyanobacteria species, specifically unicellular picocyanobacteria, under various nutritional conditions. Our experiments show that the accumulation of polyP in cyanobacteria is strongly dynamic, depending on phosphate levels and growth stages. 'Overplus' uptake of phosphorus (P) during the lag phase leads to the rapid accumulation of polyP, followed by lower polyP quotas during the exponential growth stage as a result of competing 'luxury' P uptake and polyP utilization to support rapid cell division. Cyanobacteria are capable of P deficiency responses that preferentially maintain polyP. However, preferential utilization of polyP occurs under severe P stress, suggesting the crucial role of polyP as P reserve to support cellular survival. Strong variability was observed among different species of cyanobacteria in their ability to accumulate polyP, and likely in the threshold P levels at which preferential polyP degradation occurs. This suggests that some cyanobacteria may be more adaptive to P-stressed or P-fluctuating conditions. Our results explain and provide important insights into the variability of polyP observed in aquatic environments where picocyanobacteria are the dominant primary producers.

Proto-dolomite formation in microbial consortia dominated by Halomonas strains.

Microbes can be found in hypersaline environments forming diverse populations with complex ecological interactions. Microbes in such environments were found to be involved in the formation of minerals including dolomite, a mineral of economic importance and whose origin has been long-debated. Various reports on in vitro experiments using pure cultures provided evidence for the microbial role in dolomite formation. However, culturing experiments have been limited in scope and do not fully address the possible interactions of the naturally occurring microbial communities; consequently, the ability of microbes as a community to form dolomite has been investigated in this study. Our experiments focused on examining the microbial composition by culturing aerobic heterotrophs from the top hypersaline sediments of Al-Khiran sabkha in Kuwait, a modern dolomite-forming environment. The objectives of this study were to assess the ability of two microbial consortia to form dolomite using enrichment culture experiments, mineralogy, and metagenomics. Proto-dolomite was formed by a microbial community dominated by Halomonas strains whereby degradation of the extracellular polymeric substances (EPS) was observed and the pH changed from 7.00 to 8.58. Conversely, proto-dolomite was not observed within a microbial community dominated by Clostridiisalibacter in which EPS continuously accumulated and the pH slightly changed from 7.00 to 7.29.

Microbial community composition and dolomite formation in the hypersaline microbial mats of the Khor Al-Adaid sabkhas, Qatar.

The Khor Al-Adaid sabkha in Qatar is among the rare extreme environments on Earth where it is possible to study the formation of dolomite-a carbonate mineral whose origin remains unclear and has been hypothetically linked to microbial activity. By combining geochemical measurements with microbiological analysis, we have investigated the microbial mats colonizing the intertidal areas of sabhka. The main aim of this study was to identify communities and conditions that are favorable for dolomite formation. We inspected and sampled two locations. The first site was colonized by microbial mats that graded vertically from photo-oxic to anoxic conditions and were dominated by cyanobacteria. The second site, with higher salinity, had mats with an uppermost photo-oxic layer dominated by filamentous anoxygenic photosynthetic bacteria (FAPB), which potentially act as a protective layer against salinity for cyanobacterial species within the deeper layers. Porewater in the uppermost layers of the both investigated microbial mats was supersaturated with respect to dolomite. Corresponding to the variation of the microbial community's vertical structure, a difference in crystallinity and morphology of dolomitic phases was observed: dumbbell-shaped proto-dolomite in the mats dominated by cyanobacteria and rhombohedral ordered-dolomite in the mat dominated by FAPB.

Evaluation of the "Early" Use of Albumin in Children with Extensive Burns: A Randomized Controlled Trial.

OBJECTIVE: To compare early versus delayed albumin resuscitation in children with burns in terms of clinical outcome and response. DESIGN: Randomized controlled trial. SETTING: Burn center at a tertiary care teaching hospital. PATIENTS: Forty-six children aged 1-12 years with burns greater than 15-45% total body surface area admitted within 12 hours of burn injury. INTERVENTIONS: Fluid resuscitation was based on the Parkland formula (3 mL/kg/% total body surface area), adjusted according to urine output. Patients received 5% albumin solution between 8 and 12 hours post burn in the intervention group (n = 23) and 24 hours post burn in the control group (n = 23). Both groups were assessed for reduction in crystalloid fluid infusion during resuscitation, development of fluid creep, and length of hospital stay. MEASUREMENTS AND MAIN RESULTS: There was no difference between groups regarding age, weight, sex, % total body surface area, cause of burn, or severity scores. The median crystalloid fluid volume required during the first 3 days post burn was lower in the intervention than in the control group (2.04 vs 3.05 mL/kg/% total body surface area; p = 0.025 on day 1; 1.2 vs 1.71 mL/kg/% total body surface area; p = 0.002 on day 2; and 0.82 vs 1.3 mL/kg/% total body surface area; p = 0.002 on day 3). The median urine output showed no difference between intervention and control groups (2.1 vs 2.0 mL/kg/hr; p = 0.152 on day 1; 2.58 vs 2.54 mL/kg/hr; p = 0.482 on day 2; and 2.9 vs 3.0 mL/kg/hr; p = 0.093 on day 3). Fluid creep was observed in 13 controls (56.5%) and in one patient (4.3%) in the intervention group. The median length of hospital stay was 18 days (range, 15-21 d) for controls and 14 days (range, 10-17 d) in the intervention group (p = 0.004). CONCLUSIONS: Early albumin infusion in children with burns greater than 15-45% total body surface area reduced the need for crystalloid fluid infusion during resuscitation. Significantly fewer cases of fluid creep and shorter hospital stay were also observed in this group of patients.

Impact of bacterial admixtures on the compressive and tensile strengths, permeability, and pore structure of ternary mortars: Comparative study of ureolytic and phototrophic bacteria.

The addition of bacterial biomass to cementitious materials can improve strength and permeability properties by altering the pore structure. Photoautotrophic bacteria are understudied mortar bio-additives that do not produce unwanted by-products compared to commonly studied ureolytic species. This study directly compares the impact of the addition of heterotrophic Bacillus subtilis to photoautotrophic Synechocystis sp. PCC6803 on mortar properties and microstructure. Cellulose fibers were used as a bacteria carrier. A commercial concrete healing agent composed of dormant bacterial spores was also tested. Strength, water absorption tests, mercury intrusion porosimetry, differential scanning calorimetry, thermogravimetric analysis, and scanning electron microscopy were applied to experimental mortar properties. The photoautotrophic modifications had a stronger positive impact on mortar strength and permeability properties than sporulated heterotrophic modifications due to differences in surface properties and production of exopolysaccharides. The findings provide support for photoautotrophic species as additives for mortars to move away from ammonia-generating species.

Microscopic Raman study of fungal pigment using the genetically amenable rock inhabitant Knufia petricola as a model organism.

Fungal pigments such as melanin and carotenoids are distinctive markers of animal and plant pathogenic fungi as well as their environmental relatives. These complex pigments play important roles in pathogenicity and stress tolerance while also being useful as biomarkers. Accordingly, it is important to be able to identify in situ the pigments in black fungi, a group of clinical and environmental importance. In this study, wild-type and genetically modified strains of Knufia petricola A95 and wild fungal cells attached to ancient rock were investigated for their spectroscopic and microscopic Raman features and morphological appearance. Knockout mutants of melanin synthesis genes pks1 (polyketide synthase), sdh1 (scytalone dehydratase), and both pks1 and the carotenoid synthesis gene phd1 (phytoene desaturase) were studied We applied two different Raman microscopes using two lasers, with 633 nm and 488 nm wavelengths. We analyzed and compared Raman spectra between the measured reference substances and the mutant and wild-type strains. In the wild strain WT:A95, the peaks close to melanin peals were found at 1353 cm-1 and 1611 cm-1. There are no characteristic melanin peaks at 1580-1600 cm-1 and around 1350 cm-1 at the spectrum of the Δpks1/Δphd1 mutant and the Δsdh1 mutant. The Δpks1 mutant spectrum has the peaks at the beta-carotene v2 C-C in-plane stretch at 1155 cm-1 and v3 C-CH3 deformation at 1005 cm-1. The peaks of carotenoids and melanin were found in all mutants and the wild strain, except the Δpks1/Δphd1 mutant. Raman spectra allow for discrimination between the various pigments. Hence, interactions between natural fungal melanin, as well as other protective pigments, and complex environmental matrices can be characterized on a range of spatial and temporal scales.

Raman Spectroscopic and Microbial Study of Biofilms Hosted Gypsum Deposits in the Hypersaline Wetlands: Astrobiological Perspective.

Gypsum (CaSO4·2H2O) has been identified at the surface of Mars, by both orbiters and rovers. Because gypsum mostly forms in the presence of liquid water as an essential element for sustaining microbial life and has a low porosity, which is ideal for preserving organic material, it is a promising target to look for signs of past microbial life. In this article, we studied organic matter preservation within gypsum that precipitates in a salt flat or a so-called coastal sabkha located in Qatar. Sabkha's ecosystem is considered a modern analog to evaporitic environments that may have existed on early Mars. We collected the sediment cores in the areas where gypsum is formed and performed DNA analysis to characterize the community of extremophilic microorganisms that is present at the site of gypsum formation. Subsequently, we applied Raman spectroscopy, a technique available on several rovers that are currently exploring Mars, to evaluate which organic molecules can be detected through the translucent gypsum crystals. We showed that organic material can be encapsulated into evaporitic gypsum and detected via Raman microscopy with simple, straightforward sample preparation. The molecular biology data proved useful for assessing to what extent complex Raman spectra can be linked to the original microbial community, dominated by Halobacteria and methanogenic archaea, providing a reference for a signal that may be detected on Mars.

Variability of blue carbon storage in arid evaporitic environment of two coastal Sabkhas or mudflats.

Coastal Sabkhas are mudflats found in arid coastal regions that are located within the supratidal zone when high rates of evaporation lead to high salinity. While evaporitic minerals often accumulate underneath the surface, the microbial mats are present on the surface of Sabkhas. Coastal Sabkha, an under-studied ecosystem in Qatar, has the potential to store blue carbon. In the present study, we investigated the carbon storage capacity of two Sabkhas from contrasting geological backgrounds. The spatial and temporal variabilities of the carbon stocks were examined. The results showed that both studied Sabkhas exhibit a considerable potential for soil carbon storage with carbon stocks of 109.11 ± 7.07 Mg C ha-1 and 67.77 ± 18.10 Mg C ha-1 in Dohat Faishakh and Khor al Adaid Sabkha respectively. These values fall within the reported range for carbon stocks in coastal Sabkhas in the region (51-194 Mg C ha-1). Interestingly, the carbon stocks in the sediments of the Sabkhas were higher than those in the sediments of Qatari mangroves (50.17 ± 6.27 Mg C ha-1). These finding suggest that coastal Sabkhas can serve as blue carbon ecosystems in arid environments.

Continuous Bayesian network for studying the causal links between phosphorus loading and plankton patterns in Lake Simcoe, Ontario, Canada.

An ecosystem perspective to restoring beneficial uses in Areas of Concern can be interpreted as a shift from the traditional elucidation of simple cause-effect relationships to a multicausal way of thinking that more effectively accommodates ecosystem complexity. This holistic management paradigm has also pervaded the contemporary ecological modeling practice, making compelling the adoption of more sophisticated ecosystem modeling tools. In this study, our primary objective is to develop a Bayesian hierarchical network of simple ecological models for Lake Simcoe, Ontario, Canada, aiming to establish a realistic representation of the causal connections among exogenous nutrient loading, ambient nutrient conditions, and epilimnetic plankton dynamics. In particular, we used a spatially explicit simple mass-balance model forced with idealized sinusoidal loading to predict total phosphorus concentrations. A structural equation model was then used to delineate the interplay among nutrients, ambient light conditions, phytoplankton, and herbivorous biomass. Our analysis highlights the strength of the causal linkages between total phosphorus and water clarity with phytoplankton as well as the capacity of zooplankton grazing to modulate the algal standing crop. Our Bayesian network is also used to examine the exceedance frequency of threshold values for total phosphorus (15 µg/L) and chlorophyll a (4 µg/L) concentrations under scenarios of phosphorus loading reduction. Our study suggests that a 15% phosphorus loading decrease will still result in >25% violations of the 4 µg chla/L value in the two embayments of Lake Simcoe (Cook's Bay and Kempenfelt Bay). The TP levels will decrease in response to the exogenous loading reductions and this improvement will be primarily manifested in the northcentral segments of the system.

Examining the Ability of Aerobic Halophilic Heterotrophic Microbial Consortia to Replace Ca by Mg in Different CaCO3 Precursors.

Recent laboratory experiments have exhibited microbes as promising agents in solving the perplexing origin of ancient dolomite by demonstrating microbial capability to mediate dolomite nucleation and growth. However, dolomite crystals from laboratory experiments have shown irrelevant characteristics to ancient dolomite from mineralogical and petrological perspectives. A major irrelevant characteristic is that ancient dolomite was assumed to be formed after the replacement of Ca by Mg in precursor CaCO3 in a process known as diagenesis, which contrasts with the primary precipitation process observed in laboratory culturing experiments. Considering dolomite microbial experiments, one can imply the involvement of microbes in the formation of ancient dolomite, as microbes have shown the ability to overcome the dolomite kinetic barrier. Despite that fact, the ability of microbes in mediating dolomite diagenesis has not been investigated. In this study, microbes were applied to mediate replacement of Ca by Mg in different CaCO3 precursors. The microbial replacement experiments were based on the enrichment of aerobic halophilic heterotrophic microbial consortia sampled from sediments collected from Al-Subiya sabkha in Kuwait. Two experiments were performed in saturated media at 35°C for 14 and 30 days simulating the conditions of microbial dolomite experiments. The change in mineralogy was examined via powder X-ray diffraction (XRD), and the change in texture and compositional microstructures was examined using scanning electron microscopy with energy-dispersive spectroscopy (SEM-EDS). The effect of microbes on the alteration of CaCO3 precursors was studied by comparing biotic experimentations with abiotic controls. The biotic samples were shown to result in the favorable conditions for dolomite formation including an increase in pH and alkalinity, but no changes were observed in mineralogy or compositional microstructure of CaCO3 precursors. Our results suggest the inability of aerobic halophilic heterotrophic microbial consortia to introduce Mg replacement on CaCO3 precursors in a timely manner that is comparable to primary precipitation in microbial dolomite experiments. The inability of the enriched microbial consortia to mediate replacement can be ascribed to different factors controlling the diagenetic process compared to primary precipitation in microbial dolomite experiments.

Nanoplastics: Detection and impacts in aquatic environments - A review.

The rise in the global production of plastics has led to severe concerns about the impacts of plastics in aquatic environments. Although plastic materials degrade over extreme long periods, they can be broken down through physical, chemical, and/or biological processes to form microplastics (MPs), defined here as particles between 1 µm and 5 mm in size, and later to form nanoplastics (NPls), defined as particles <1 µm in size. We know little about the abundance and effects of NPls, even though a lot of research has been conducted on the ecotoxicological impacts of MPs on both aquatic biota. Nevertheless, there is evidence that NPls can both bypass the cell membranes of microorganisms and bioaccumulate in the tissues and organs of higher organisms. This review analyzes 150 publications collected by searching through the databases Web of Science, SCOPUS, and Google Scholar using keywords such as nanoplastics*, aquatic*, detection*, toxic*, biofilm*, formation*, and extracellular polymeric substance* as singular or plural combinations. We highlight and critically synthesize current studies on the formation and degradation of NPls, NPls' interactions with aquatic biota and biofilm communities, and methods of detection. One reason for the missing data and studies in this area of research is the lack of a protocol for the detection of, and suitable methods for the characterization of, NPls in the field. Our primary aim is to identify gaps in knowledge throughout the review and define future directions of research to address the impacts of NPls in aquatic environments. The development of consistent and standardized sets of procedures would address the gaps in knowledge regarding the formation and degradation of NPls as well as sampling and characterizing natural NPls needed to observe the full extent of NPls on aquatic biota and biofilm communities.

Silicon: The key element in early stages of biocalcification.

Biocalcification is a widespread process of forming hard tissues like bone and teeth in vertebrates. It is also a topic connecting life sciences and earth sciences: calcified skeletons and shells deposited as sediments represent the earth's fossil record and are of paramount interest for biogeochemists trying to get an insight into the past of our planet. This study reports on the role of silicon in the early biocalcification steps, where silicon and calcium were detected on the surface of cyanobacteria (initial stage of lacustrine calcite precipitation) and in crustacean cuticles. By using innovative methodological approaches of correlative microscopy (AFM in combination with analytical TEM: EFTEM, EELS) the chemical form of silicon in biocalcifying matrices and organic-inorganic particles is determined. Previously, silicon was reported to be localized in active growth areas in the young bone of vertebrates. We have found evidence that biocalcification in evolutionarily distant organisms involves very similar initial phases with silicon as a key element at the organic-inorganic interface.

Dolomite genesis in bioturbated marine zones of an early-middle Miocene coastal mud volcano outcrop (Kuwait).

The origin of spheroidal dolomitized burrow from Al-Subiya sabkha in Kuwait was previously described as enigmatic as no evidence of precursor calcium carbonate was found in the siliciclastic sediment. An assumption for the genesis of spheroidal dolomite from the same area was attributed to hydrocarbon seepage but no evidence was provided. Here, we investigated a recently discovered early-middle Miocene coastal mud volcano outcrop in Al-Subiya sabkha where dolomitized burrows and spheroidal dolomite are found in bioturbated marine zones, and associated with traces of salt. Conversely, the continental zone lacks bioturbation features, dolomite and traces of salt, which together contrast with bioturbated rich marine zones. Geochemical signatures of Rare Earth Elements + Yttrium show a true positive Ce anomaly (Ce/Ce* > 1.2) and positive Eu/Eu* anomaly of spheroidal dolomite indicating strictly anoxic conditions, and sulphate reduction to sulphide, respectively. Our results are suggestive of a relationship between dolomite formation and interdependent events of hydrocarbon seepage, flux of hypersaline seawater, bioturbation, and fluid flow in the marine zones of the mud volcano. The bioturbation activity of crustaceans introduced channels/burrows in the sediment-water interface allowing for the mixing of seeped pressurized hydrocarbon-charged fluids, and evaporitic seawater. In the irrigated channels/burrows, the seeped pressurized hydrocarbon-charged fluids were oxidized via microbial consortia of methanotrophic archaea and sulphate-reducing bacteria resulting in elevated alkalinity and saturation index with respect to dolomite, thus providing the preferential geochemical microenvironment for dolomite precipitation in the bioturbated sediment.

Environmentally friendly antibiofilm strategy based on cationized phytoglycogen nanoparticles.

Biofilm tolerance to antibiotics has led to the search for new alternatives in treating biofilms. The use of metallic nanoparticles has been a suggested strategy against biofilms, but their potential environmental toxicity and high cost of synthesizing have limited their applications. In this study, we investigate the potential of polysaccharidic phytoglycogen nanoparticles extracted from corn, in treating cyanobacterial biofilms, which are the source of toxins and pollution in aquatic environments. Our results revealed that the surface of cyanobacterial cells was dominated by the negatively charged functional groups such as carboxylic and phosphoric groups. The native phytoglycogen (PhX) nanoparticles were dominated with non-charged groups, such as hydroxyl groups, and the cationized phytoglycogen (PhXC) nanoparticles showed positively charged surfaces due to the presence of quaternary ammonium cations. Our results indicated that, as opposed to PhX, PhXC strongly inhibited biofilm formation when dispersed in the culture medium. PhXC also eradicated the already grown cyanobacterial biofilms. The antibiofilm properties of PhXC were attributed to its strong electrostatic interactions with the cyanobacterial cells, which could inhibit cell/cell and cell/substrate interactions and nutrient exchange with the media. This class of antibacterial polysaccharide nanoparticles may provide a novel cost-effective and environment-friendly strategy for treating biofilm formation by a broad spectrum of bacteria.

Attachment on mortar surfaces by cyanobacterium Gloeocapsa PCC 73106 and sequestration of CO2 by microbially induced calcium carbonate.

Cyanobacterial carbonate precipitation induced by cells and extracellular polymeric substances (EPS) enhances mortar durability. The percentage of cell/EPS attachment regulates the effectiveness of the mortar restoration. This study investigates the cell coverage on mortar and microbially induced carbonate precipitation. Statistical analysis of results from scanning electron and fluorescence microscopy shows that the cell coverage was higher in the presence of UV-killed cells than living cells. Cells are preferably attached to cement paste than sand grains, with a difference of one order of magnitude. The energy-dispersive X-ray spectroscopy analyses and Raman mapping suggest cyanobacteria used atmospheric CO2 to precipitate carbonates.

Modern dolomite formation caused by seasonal cycling of oxygenic phototrophs and anoxygenic phototrophs in a hypersaline sabkha.

The "Dolomite Problem" has been a controversy for over a century, owing to massive assemblages of low-temperature dolomite in ancient rocks with little dolomite forming today despite favorable geochemical conditions. Experiments show that microbes and their exopolymeric substances (EPS) nucleate dolomite. However, factors controlling ancient abundances of dolomite can still not be explained. To decode the enigma of ancient dolomite, we examined a modern dolomite forming environment, and found that a cyclic shift in microbial community between cyanobacteria and anoxygenic phototrophs creates EPS suited to dolomite precipitation. Specifically, EPS show an increased concentration of carboxylic functional groups as microbial composition cycles from cyanobacterial to anoxygenic phototroph driven communities at low-and high- salinity, respectively. Comparing these results to other low-T forming environments suggests that large turnover of organic material under anoxic conditions is an important driver of the process. Consequently, the shift in atmospheric oxygen throughout Earth's history may explain important aspects of "The Dolomite Problem". Our results provide new context for the interpretation of dolomite throughout Earth's history.

Systematic laboratory approach to produce Mg-rich carbonates at low temperature.

Dolomite is a common Mg-rich carbonate in the geological record, but the mechanism of its formation remains unclear. At low temperature, the incorporation of magnesium ions into the carbonate minerals necessary to form dolomite is kinetically inhibited. Over the decades, several factors that possibly allow for overcoming this kinetic barrier have been proposed, and their effectiveness debated. Here, we present the results of a large number of laboratory precipitation experiments that have been designed to identify and compare the factors that promote the formation of Mg-rich carbonates. Under the tested conditions, the most interesting observations include: (1) from solutions that mimic evaporitic seawater, the maximum mol% of Mg incorporated in high Mg calcite is 35, (2) carbonates with a mol% of Mg above 40 were obtained exclusively in the presence of organic molecules, (3) no correlation was observed between the charge of the organic molecules and the incorporation of Mg, (4) the mode (i.e., slow vs. fast mixing) used to add carbonate to the solution obtaining supersaturation has a significant impact on the forming mineral phase (aragonite vs. nesquehonite vs. high Mg calcite) and its Mg content. These findings allow for a more informed evaluation of the existing models for dolomite formation, which are based on the study of natural environments and ancient sedimentary sequences.