Nutrient dynamics and microbial community response in macrophyte-dominated lakes: Implications for improved restoration strategies.

Although lakes dominated by macrophytes are conducive to ecological balance, this balance is easily disrupted by excessive nutrients flowing into the lake. However, knowledge of whether excessive nutrients lead to different microbial environmental vulnerabilities in the lake sediment between macrophyte-dominated areas and macrophyte-free areas is a prerequisite for the implementation of targeted protection measures. In this study, we investigated bacterial communities in sediments using high-throughput sequencing of 16S rRNA genes. Our results showed that the sources of total nitrogen (TN) and organic matter (OM) were related to the macrophytes. The structure, drivers, and interspecific associations of bacterial community, which were more susceptible to increased changes in TN and OM, differed significantly between macrophyte-dominated areas and macrophyte-free areas. More precisely, the lake edge, where was occupied by macrophytes, had a higher proportion of deterministic phylogenetic turnover (88.89%) than other sites, as well as a wider ecological niche and a tighter network structure. Further, as the difference in TN increased, the main assembly processes in surface sediments changed from stochastic to deterministic. However, the majority of phyla from the lake edge showed a greater correlation with excessive nutrients, and the selection of the community by excessive nutrients was more obvious at the edge of the lake. In addition, our results demonstrated that the stability of the bacterial community in macrophyte-free areas is greater than in macrophyte-dominated areas, while an excessively high deterministic process ratio and nutrient (TN and OM) concentration significantly reduced bacterial community stability at macrophyte-dominated areas. Taken together, these results provide a better understanding of the effects of excessive nutrients derived from macrophytes on bacterial community patterns, and highlight the importance of avoiding the accumulation of TN and OM in macrophyte-dominated areas to enhance the sustainability of the ecosystem after restoration of lakes with macrophytes.

Probing the metabolic water contribution to intracellular water using oxygen isotope ratios of PO4.

Knowledge of the relative contributions of different water sources to intracellular fluids and body water is important for many fields of study, ranging from animal physiology to paleoclimate. The intracellular fluid environment of cells is challenging to study due to the difficulties of accessing and sampling the contents of intact cells. Previous studies of multicelled organisms, mostly mammals, have estimated body water composition-including metabolic water produced as a byproduct of metabolism-based on indirect measurements of fluids averaged over the whole organism (e.g., blood) combined with modeling calculations. In microbial cells and aquatic organisms, metabolic water is not generally considered to be a significant component of intracellular water, due to the assumed unimpeded diffusion of water across cell membranes. Here we show that the (18)O/(16)O ratio of PO4 in intracellular biomolecules (e.g., DNA) directly reflects the O isotopic composition of intracellular water and thus may serve as a probe allowing direct sampling of the intracellular environment. We present two independent lines of evidence showing a significant contribution of metabolic water to the intracellular water of three environmentally diverse strains of bacteria. Our results indicate that ∼30-40% of O in PO4 comprising DNA/biomass in early stationary phase cells is derived from metabolic water, which bolsters previous results and also further suggests a constant metabolic water value for cells grown under similar conditions. These results suggest that previous studies assuming identical isotopic compositions for intracellular/extracellular water may need to be reconsidered.

Oxygen isotope studies of phosphite oxidation: purification and analysis of reactants and products by high-temperature conversion elemental analyzer/isotope ratio mass spectrometry.

RATIONALE: Increased attention has been recently focused on the origin and reactions of reduced-P oxyanions such as phosphite [PO3 (III)] in terrestrial and biological systems. We present new methods for studying O-isotopic reactions between PO3 (III) and other oxygen sources during oxidation of PO3 (III) to PO4 (V). METHODS: Na2 (HPO3 )·5H2 O, used as a PO3 (III) source, contains structural water due to its hygroscopic nature; thus, we developed a method for determining the δ(18) O value of PO3 (III) after the removal of structural water. Next, we tested two techniques for purifying PO4 (V) from aqueous PO3 (III)/PO4 (V) mixtures: (1) precipitation of PO4 (V) as ammonium phosphomolybdate (APM); and (2) precipitation of PO4 (V) as magnesium ammonium phosphate (MAP). The O-isotope compositions, (18) O:(16) O (δ(18) O values), of Na2 (HPO3 ) and Ag3 PO4 were analyzed by TC/EA/IRMS. RESULTS: Structural water was removed from Na2 (HPO3 )·5H2 O after drying at 100 °C under vacuum and the δ(18) O value of PO3 (III) was obtained. The δ(18) O values of PO4 (V), which was extracted from (18) O-labeled PO3 (III)/PO4 (V) mixtures by APM and MAP precipitations, were not altered by the precipitation process. This result confirms that PO3 (III) is not converted into PO4 (V) by oxidation or hydrolysis under either strong acidic/oxidizing (APM) or alkaline (MAP) conditions for up to a 24-h period. CONCLUSIONS: We conclude that both APM and MAP precipitation are reliable and effective methods for the separation and purification of PO4 (V) from aqueous PO3 (III)/PO4 (V) mixtures. The methods described here will permit the study of the isotopic evolution of various pathways of geochemical as well as biological PO3 (III) oxidation.

Functional gene expression of oil-degrading bacteria resistant to hexadecane toxicity.

Contamination with oil poses a threat to the environment and to human health worldwide. Biological methodologies have proved to be economical, versatile and efficient for the remediation of pollutants. In this paper, a highly efficient oil-degrading bacterial strain USTB-2 was isolated from an oil production well of Dagang oil field in Tianjin, China. The 16S rRNA sequence of USTB-2 showed 100% similarity with that of Bacillus subtilis BSn5. Hexadecane is one of the most important components in petroleum. The half inhibitory ratio (IC50) of hexadecane inhibited organisms, determined by microcalorimetry, was lower in USTB-2 than in B. BSn5. The results indicate that the strain USTB-2 degrades hexadecane to make it less toxic compared with the normal strain. RT-PCR was used to evaluate the expression of oil-degrading enzymes, specifically 4-hydroxyphenylacetate 3-monooxygenase genes (HPMO). A sharp increase in the expression of HPMO genes was observed for USTB-2, while the expression of HPMO genes in reference strain B. BSn5 remained relatively stable. These methods can be used to study the metabolic potential of microorganisms for in situ oil decontamination.

Oxygen isotope signature of UV degradation of glyphosate and phosphonoacetate: tracing sources and cycling of phosphonates.

The degradation of phosphonates in the natural environment constitutes a major route by which orthophosphate (Pi) is regenerated from organic phosphorus and recently implicated in marine methane production, with ramifications to environmental pollution issues and global climate change concerns. This work explores the application of stable oxygen isotope analysis in elucidating the CP bond cleavage mechanism(s) of phosphonates by UV photo-oxidation and for tracing their sources in the environment. The two model phosphonates used, glyphosate and phosphonoacetic acid were effectively degraded after exposure to UV irradiation. The isotope results indicate the involvement of both ambient water and atmospheric oxygen in the CP bond cleavage and generally consistent with previously posited mechanisms of UV-photon excitation reactions. A model developed to calculate the oxygen isotopic composition of the original phosphonate P-moiety, shows both synthetic phosphonates having distinctly lower values compared to naturally derived organophosphorus compounds. Such mechanistic models, based on O-isotope probing, are useful for tracing the sources and reactions of phosphonates in the environment.

Measurement of δ18O values in arsenic and selenium oxyanions.

RATIONALE: Past stable isotopic studies of selenium and arsenic oxyanions within a biogeochemistry context have focused solely on selenium isotopes due to monoisotopic nature of arsenic and lack of attention to oxygen. A modern quantification method of δ(18)O values in selenate, selenite, arsenate, and arsenite is presented here, similar to methods for other inorganic oxyanions such as phosphate. METHODS: Dissolved oxyanions were recovered after precipitation with silver or barium cations within controlled pH ranges and purified by drying and roasting under vacuum. Oxygen isotope ratio measurements were performed on 250-450 micrograms of solids using continuous flow high-temperature conversion elemental analyzer/isotope ratio mass spectrometry (CF-TCEA/IRMS). Each Se and As solid was evaluated for reproducibility and stability of δ(18)O values and the usefulness of this method for evaluating aqueous phase Se and As oxyanion reactions was addressed. RESULTS: Only silver arsenate and barium selenate were shown to have stable, reproducible δ(18)O values over 5 months and were calibrated to the VSMOW scale. The method was applied to confirm complete exchange of oxygen isotopes between arsenate and solvent water at pH 7 which produced an O-isotope fractionation between arsenate and water of 6.0‰. Furthermore, the method revealed the ability of oxygen in selenate to kinetically fractionate during reduction to elemental selenium by a green rust compound which produced a fractionation (20.9‰) larger than reported δ(80/76)Se values for similar reactions. CONCLUSIONS: The experimental and analytical conditions for δ(18)O analysis of Se and As oxyanions were detailed. δ(18)O analyses of As and Se oxyanions may be useful in biogeochemical studies for tracing chemical reaction pathways, determining environmental sources of Se, or diagnosing specific reaction mechanisms for As and Se oxyanions.

Effects of petroleum contamination on soil microbial numbers, metabolic activity and urease activity.

The influence of petroleum contamination on soil microbial activities was investigated in 13 soil samples from sites around an injection water well (Iw-1, 2, 3, 4) (total petroleum hydrocarbons (TPH): 7.5-78 mg kg(-1)), an oil production well (Op-1, 2, 3, 4, 5) (TPH: 149-1110 mg kg(-1)), and an oil spill accident well (Os-1, 2, 3, 4) (TPH: 4500-34600 mg kg(-1)). The growth rate constant (µ) of glucose stimulated organisms, determined by microcalorimetry, was higher in Iw soil samples than in Op and Os samples. Total cultivable bacteria and fungi and urease activity also decreased with increasing concentration of TPH. Total heat produced demonstrated that TPH at concentrations less than about 1 g kg(-1) soil stimulated anaerobic respiration. A positive correlation between TPH and soil organic matter (OM) and stimulation of fungi-bacteria-urease at low TPH doses suggested that TPH is bound to soil OM and slowly metabolized in Iw soils during OM consumption. These methods can be used to evaluate the potential of polluted soils to carry out self-bioremediation by metabolizing TPH.

Biotic and abiotic pathways of phosphorus cycling in minerals and sediments: insights from oxygen isotope ratios in phosphate.

A key question to address in the development of oxygen isotope ratios in phosphate (δ(18)O(p)) as a tracer of biogeochemical cycling of phosphorus in ancient and modern environments is the nature of isotopic signatures associated with uptake and cycling of mineral-bound phosphate by microorganisms. Here, we present experimental results aimed at understanding the biotic and abiotic pathways of P cycling during biological uptake of phosphate sorbed to ferrihydrite and the selective uptake of sedimentary phosphate phases by Escherichia coli and Marinobacter aquaeolei. Results indicate that a significant fraction of ferrihydrite-bound phosphate is biologically available. The fraction of phosphate taken up by E. coli attained an equilibrium isotopic composition in a short time (<50 h) due to efficient O-isotope exchange (between O in PO(4) and O in water; that is, actual breaking and reforming of P-O bonds) (biotic pathway). The difference in isotopic composition between newly equilibrated aqueous and residual sorbed phosphate groups promoted the ion exchange (analogous to isotopic mixing) of intact phosphate ions (abiotic pathway) so that this difference gradually became negligible. In sediment containing different P phases, E. coli extracted loosely sorbed phosphate first, whereas M. aquaeolei preferred Fe-oxide-bound phosphate. The presence of bacteria always imprinted a biotic isotopic signature on the P phase that was taken up and cycled. For example, the δ(18)O(p) value of loosely sorbed phosphate shifted gradually toward equilibrium isotopic composition. The δ(18)O(p) value of Fe-oxide-bound phosphate, however, showed only slight changes initially but, when new Fe-oxides were formed, coprecipitated/occluded phosphate retained δ(18)O values of the aqueous phosphate at the time of formation of new Fe oxides. Concentrations and isotopic compositions of authigenic and detrital phosphates did not change, suggesting that these phosphate phases were not utilized by bacteria. These findings support burgeoning applications of δ(18)O(p) as a tracer of phosphorus cycling in sediments, soils, and aquatic environments and as an indicator of paleo- environmental conditions.

Phosphate oxygen isotopic evidence for a temperate and biologically active Archaean ocean.

Oxygen and silicon isotope compositions of cherts and studies of protein evolution have been interpreted to reflect ocean temperatures of 55-85 degrees C during the early Palaeoarchaean era ( approximately 3.5 billion years ago). A recent study combining oxygen and hydrogen isotope compositions of cherts, however, makes a case for Archaean ocean temperatures being no greater than 40 degrees C (ref. 5). Ocean temperature can also be assessed using the oxygen isotope composition of phosphate. Recent studies show that (18)O:(16)O ratios of dissolved inorganic phosphate (delta(18)O(P)) reflect ambient seawater temperature as well as biological processing that dominates marine phosphorus cycling at low temperature. All forms of life require and concentrate phosphorus, and as a result of biological processing, modern marine phosphates have delta(18)O(P) values typically between 19-26 per thousand (VSMOW), highly evolved from presumed source values of approximately 6-8 per thousand that are characteristic of apatite in igneous rocks and meteorites. Here we report oxygen isotope compositions of phosphates in sediments from the 3.2-3.5-billion-year-old Barberton Greenstone Belt in South Africa. We find that delta(18)O(P) values range from 9.3 per thousand to 19.9 per thousand and include the highest values reported for Archaean rocks. The temperatures calculated from our highest delta(18)O(P) values and assuming equilibrium with sea water with delta(18)O = 0 per thousand (ref. 12) range from 26 degrees C to 35 degrees C. The higher delta(18)O(P) values are similar to those of modern marine phosphate and suggest a well-developed phosphorus cycle and evolved biologic activity on the Archaean Earth.

Microbial diversity of boron-rich volcanic hot springs of St. Lucia, Lesser Antilles.

The volcanic Sulphur Springs, St. Lucia, present an extreme environment due to high temperatures, low pH values, and high concentrations of sulfate and boron. St. Lucia offers some unique geochemical characteristics that may shape the microbial communities within the Sulphur Springs area. We chose six pools representing a range of geochemical characteristics for detailed microbial community analyses. Chemical concentrations varied greatly between sites. Microbial diversity was analyzed using 16S rRNA gene clone library analyses. With the exception of one pool with relatively low concentrations of dissolved ions, microbial diversity was very low, with Aquificales sequences dominating bacterial communities at most pools. The archaeal component of all pools was almost exclusively Acidianus spp. and did not vary between sites with different chemical characteristics. In the pool with the highest boron and sulfate concentrations, only archaeal sequences were detected. Compared with other sulfur springs such as those at Yellowstone, the microbial diversity at St. Lucia is very different, but it is similar to that at the nearby Lesser Antilles island of Montserrat. While high elemental concentrations seem to be related to differences in bacterial diversity here, similarities with other Lesser Antilles sites suggest that there may be a biogeographical component as well.

Transport of single-walled carbon nanotubes in porous media: filtration mechanisms and reversibility.

Deposition of nanomaterials onto surfaces is a key process governing their transport, fate, and reactivity in aquatic systems. We evaluated the transport and deposition behavior of carboxyl functionalized single-walled carbon nanotubes (SWNTs) in a well-defined porous medium composed of clean quartz sand over a range of solution chemistries. Our results showthat increasing solution ionic strength or addition of calcium ions result in increased SWNT deposition (filtration). This observation is consistent with conventional colloid deposition theories, thereby suggesting that physicochemical filtration plays an important role in SWNT transport. However, the relatively insignificant change of SWNT filtration at low ionic strengths (< or = 3.0 mM KCl) and the incomplete breakthrough of SWNTs in deionized water (C/Co = 0.90) indicate that physical straining also plays a role in the capture of SWNTs within the packed sand column. It is proposed that SWNT shape and structure, particularly the very large aspect ratio and its highly bundled (aggregated) state in aqueous solutions, contribute considerably to straining in flow through porous media. We conclude that both physicochemical filtration and straining play a role at low (< 3.0 mM) ionic strength, while physicochemical filtration is the dominant mechanism of SWNT filtration at higher ionic strengths. Our results further show that deposited SWNTs are mobilized (released) from the quartz sand upon introduction of low ionic strength solution following deposition experiments with monovalent salt (KCl). In contrast, SWNTs deposited in the presence of calcium ions were not released upon introduction of low ionic strength solution to the packed column, even when humic acid was present in solution during SWNT deposition.

Marine phosphate oxygen isotopes and organic matter remineralization in the oceans.

We show that the isotopic composition of oxygen (delta18O) in dissolved inorganic phosphate (Pi) reveals the balance between Pi transport and biological turnover rates in marine ecosystems. Our delta18Op of Pi (delta18Op) measurements herein indicate the importance of cell lysis in the regeneration of Pi in the euphotic zone. Depth profiles of the delta18Op in the Atlantic and Pacific Oceans are near a temperature-dependent isotopic equilibrium with water. Small deviations from equilibrium below the thermocline suggest that P remineralization in the deep ocean is a byproduct of microbial carbon and energy requirements. However, isotope effects associated with phosphohydrolase enzymes involved in P remineralization are quite large and could potentially lead to significant disequilibration of Pi oxygen. The observed near equilibration of deep water Pi likely calls for continued slow rates of microbial uptake and release of Pi and/or extracellular pyrophosphatase-mediated oxygen exchange between water and Pi along the deep water flow path.