Adaptive responses of aglomerular toadfish to dilute sea water

Plasma and urine of toadfish (Opsanus tau) in sea water and 10% sea water were analyzed to assess responses of an aglomerular fish to hypoosmotic challenge. Following transfer to 10% sea water, plasma osmotic pressure decreased slowly from 318 to 241 mmol · kg H₂O⁻¹, over a period of 10–15 days. Urine osmotic pressure decreased in parallel from 299 to 207 mmol · kg H₂O⁻¹, leaving urine/plasma ratios of osmotic pressure essentially unchanged. In contrast, the volume and composition of urine changed rapidly following transfer to 10% sea water. Urine flow rate increased 110% from 3.0 to 6.3 μl · 100g⁻¹ · h⁻¹ and Na⁺ excretion increased 346%, while excretion of Mg²⁻ and SO₄ ²⁻ decreased 81% and 90%, respectively. Excretion rates for Cl⁻ were low in seawater toadfish and decreased further in 10% sea water. An unknown sulfur-containing anion, present in the urine of seawater toadfish, contributed significantly to the composition and ionic balance in urine of toadfish in 10% sea water. These results suggest that the inability to produce strongly dilute urine obliges toadfish to lose salt in order to excrete water, in hypoosmotic media. The decrease in plasma osmotic pressure may be both a strategy to reduce osmotic and ionic gradients in dilute media and a consequence of the kidney's inability to excrete water without salt. Accepted: 22 August 1996     

Fractionation of sulfur isotopes during dissimilatory reduction of sulfate by a thermophilic gram-negative bacterium at 60 °C

Sulfur isotope (³⁴S/³²S) fractionation during reduction of dissolved sulfate was investigated with a growing batch culture of a thermophilic, gram-negative, sulfate-reducing bacterium (strain MT-96) at 60 °C. The completely oxidizing strain was isolated from geothermally heated sediments of a shallow-water hydrothermal vent in the Mediterranean Sea. The hydrogen sulfide produced in the experiments was enriched in ³²S by approximately 19‰ as compared to sulfate, which indicates that stable isotope discrimination by this thermophile is within the range found previously for mesophilic sulfate-reducing bacteria, and only slightly higher than that observed for the thermophilic gram-positive Desulfotomaculum nigrificans. Received: 1 December 1998 / Accepted: 25 May 1999     

Dissolved and particulate organic matter in contrasting Zostera marina (eelgrass) sediments

The influence of seagrass Zostera marina on sediment characteristics was examined in two contrasting sediments, one organic-rich and one organic-poor. The presence of plants leads to reduced sediment redox potential in both sediment types compared to bare sediment with the largest effects in the organic-poor sediment. Z. marina stimulated the sulfate reduction rates in organic-poor sediment with ∼50% and higher pools of dissolved organic carbon (DOC) were found. In contrast, sulfate reduction rates were lower in vegetated compared to bare sites in the organic-rich sediment. Despite a low contribution of dissolved carbohydrate (DCHO) to the DOC pool (<5%), the seagrass vegetation was responsible for an increase of ∼50% in DCHO pools with a peak in the root zone suggesting that Z. marina supplied DCHO to the pore waters. The Z. marina meadows also enhanced the contribution of particulate carbohydrate (PCHO) to sedimentary particulate organic carbon (POC) pools by 6-14% compared to bare sediment. Although the PCHO pools were higher in organic-rich than organic-poor sediments, the analyses of carbohydrate composition revealed that three groups of neutral sugars including glucose, galactose and mannose+xylose were the major compounds of PCHO and contributed with >60% to sedimentary carbohydrate pools at both sites. Only glucose showed depletion with depth in the vegetated sediments, whereas the percentage of ribose and rhamnose increased indicating a selective degradation of labile carbohydrates in the meadows. Galactose and mannose+xylose appeared to represent a refractory part of carbohydrate that remained after degradation of the more labile components. The sugar content was rather constant with depth at the bare organic-rich sediment indicating that only recalcitrant carbohydrate pools were buried. There was less difference in the PCHO composition profiles between vegetated and bare organic-poor sediments.     

Enhanced anaerobic biodegradation of nonylphenol ethoxylates by introducing additional sulfate or nitrate as terminal electron acceptors

The potential to enhance the anaerobic biodegradation of nonylphenol ethoxylates (NPEOs) by introducing additional sulfate or nitrate as electron acceptor was investigated. The results showed that adding nitrate or sulfate could significantly enhance the anaerobic biodegradation of NPEOs and alleviate the accumulation of their estrogenic intermediates. However, these terminal electron acceptors had no influence on the component of the anaerobic biodegradation products of NPEOs. To the best of our knowledge, it is the first report of the enhancement of anaerobic biodegradation of NPEOs by introducing additional terminal electron acceptor with relatively high redox potential. These observations have significant environmental implications in terms of the environmental behavior of NPEO contaminants in natural environment.     

Identification and biochemical analysis of a homolog of a sulfate transporter from a vanadium-rich ascidian Ascidia sydneiensis samea

Background

Several species of ascidians accumulate extremely high levels of vanadium ions in the vacuoles of their blood cells (vanadocytes). The vacuoles of vanadocytes also contain many protons and sulfate ions. To maintain the concentration of sulfate ions, an active transporter must exist in the blood cells, but no such transporter has been reported in vanadium-accumulating ascidians.

Methods

We determined the concentration of vanadium and sulfate ions in the blood cells (except for the giant cells) of Ascidia sydneiensis samea. We cloned cDNA for an Slc13-type sulfate transporter, AsSUL1, expressed in the vanadocytes of A. sydneiensis samea. The synthetic mRNA of AsSUL1 was introduced into Xenopus oocytes, and its ability to transport sulfate ions was analyzed.

Results

The concentrations of vanadium and sulfate ions in the blood cells (except for the giant cells) were 38 mM and 86 mM, respectively. The concentration of sulfate ions in the blood plasma was 25 mM. The transport activity of AsSUL1 was dependent on sodium ions, and its maximum velocity and apparent affinity were 2500 pmol/oocyte/h and 1.75 mM, respectively.

General significance

This could account for active uptake of sulfate ions from blood plasma where sulfate concentration is 25 mM, as determined in this study.     

Anaerobic degradation of citrate under sulfate reducing and methanogenic conditions

Citrate is an important component of metal processing effluents such as chemical mechanical planarization wastewaters of the semiconductor industry. Citrate can serve as an electron donor for sulfate reduction applied to promote the removal of metals, and it can also potentially be used by methanogens that coexist in anaerobic biofilms. The objective of this study was to evaluate the degradation of citrate with sulfate-reducing and methanogenic biofilms. During batch bioassays, the citrate, acetate, methane and sulfide concentrations were monitored. The results indicate that independent of the biofilm or incubation conditions used, citrate was rapidly fermented with specific rates ranging from 566 to 720 mg chemical oxygen demand (COD) consumed per gram volatile suspended solids per day. Acetate was found to be the main fermentation product of citrate degradation, which was later degraded completely under either methanogenic or sulfate reducing conditions. However, if either sulfate reduction or methanogenesis was infeasible due to specific inhibitors (2-bromoethane sulfonate), absence of sulfate or lack of adequate microorganisms in the biofilm, acetate accumulated to levels accounting for 90–100% of the citrate-COD consumed. Based on carbon balances measured in phosphate buffered bioassays, acetate, CO₂ and hydrogen are the main products of citrate fermentation, with a molar ratio of 2:2:1 per mol of citrate, respectively. In bicarbonate buffered bioassays, acetogenesis of H₂ and CO₂ increased the yield of acetate. The results taken as a whole suggest that in anaerobic biofilm systems, citrate is metabolized via the formation of acetate as the main metabolic intermediate prior to methanogenesis or sulfate reduction. Sulfate reducing consortia must be enriched to utilize acetate as an electron donor in order to utilize the majority of the electron-equivalents in citrate.     

Are bioleaching rates determined by the available particle surface area concentration?

It is well known that pulp density and particle size determine the available surface area concentration and have an influence in the overall rate of bioleaching of minerals. As metal solubilization takes place through the surface area of the particles, it can be expected that different combinations of pulp densities and particle sizes giving the same surface area concentration would determine the same leaching rate. The objective of this work was to test this hypothesis on the effect of surface area concentration, pulp density and particle size of the biooxidation of a pyritic gold concentrate by the thermophilic Archaeon Sulfolobus metallicus in shake flasks. The gold concentrate was used at 2.5%, 5%, 10%, and 15% w/v pulp density and at four size fractions: 150–106, 106–75, 75–38 and –38 μm. Temperature was 68°C and the initial pH was 2.0. Results showed that the volumetric productivities of iron and sulfate depend not only on the surface area concentration but also on pulp density and particle size considered separately. These two variables not only determine surface area but also exert additional effects on the process, so the hypothesis was not confirmed. Maximum attained iron productivity was 1.042 g/l day with the 75–38 μm fraction at 5% pulp density. Maximum sulfate productivity was 4.279 g/l day with the 75–38 μm fraction at 10% pulp density.     

Sulfate Activation Enzymes: Phylogeny and Association with Pyrophosphatase

The enzymes catalyzing the first two reactions in the sulfate activation pathway, ATP-sulfurylase (S) and APS-kinase (K), are fused as ‘KS’ in animals but are fused as ‘SK’ in select bacteria and fungi. We have discovered a novel triple fusion protein of K, S, and pyrophosphatase (P) in several protozoan genomes within the Stramenopile lineage. These triple domain fusion proteins led us to hypothesize that pyrophosphatase enzymes and sulfate activation enzymes physically interact to impact the thermodynamics of the sulfate activation pathway. In support of this hypothesis, we demonstrate through biochemical assays that separately encoded KS and P proteins physically interact and that KS/P complexes activate more sulfate than KS alone. We also conclude on the basis of phylogenetic analyses that all known KS fusions originate from a single fusion event early in the eukaryotic lineage. Strikingly, our analyses support the same conclusion for all known SK fusions. These observations indicate that the patchwork of fused and nonfused S and K genes observed in modern-day eukaryotes and prokaryotes are the result of the two ancestral fusion genes evolving by an assortment of gene fissions, duplications, deletions, and horizontal transfers in different lineages. Our integrative use of genomics, phylogenetics, and biochemistry to characterize pyrophosphatase as a new member of the sulfate activation pathway should be effective at identifying new protein members and connections in other molecular pathways. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users. Nucleotide sequence data reported here are available in the GenBank/DDBJ/EMBL databases under accession number EU352210.