Inhibition of the biosynthesis ofN-acetylneuraminic acid by metal ions and seleniumin vitro

In liver homogenate the biosynthesis ofN-acetylneuraminic acid usingN-acetylglucosamine as precursor can be followed stepwise by applying different chromatographic procedures. In this cell-free system 16 metal ions (Zn²⁺, Mn²⁺, La³⁺, Co²⁺, Cu²⁺, Hg²⁺, VO ₃ ^– , Pb²⁺, Ce³⁺, Cd²⁺, Fe²⁺, Fe³⁺, Al³⁺, Sn²⁺, Cs⁺ and Li⁺) and the selenium compounds, selenium(IV) oxide and sodium selenite, have been checked with respect to their ability to influence a single or possible several steps of the biosynthesis ofN-acetylneuraminic acid. It could be shown that the following enzymes are sensitive to these metal ions (usually applied at a concentration of 1 mmoll^–1):N-acetylglucosamine kinase (inhibited by Zn²⁺ and vandate), UDP-N-acetylglucosamine-2-epimerase (inhibited by zn²⁺, Co²⁺, Cu²⁺, Hg²⁺, VO ₃ ^– , Pb²⁺, Cd²⁺, Fe³⁺, Cs⁺, Li⁺, selenium(IV) oxide and selenite), andN-acetylmannosamine kinase (inhibited by Zn²⁺, Cu²⁺, Cd²⁺, and Co²⁺). Dose dependent measurements have shown that Zn²⁺, Cu²⁺ and selenite are more efficient inhibitors of UDP-N-acetylglucosamine-2-epimerase than vanadate. As for theN-acetylmannosamine kinase inhibition, a decreasing inhibitory effect exists in the following order Zn²⁺, Cd²⁺, Co²⁺ and Cu²⁺. In contrast, La³⁺, Al³⁺ and Mn²⁺ (1 mmoll^–1) did not interfere with the biosynthesis ofN-acetylneuraminic acid. Thus, the conclusion that the inhibitory effect of the metal ions investigated cannot be regarded as simply unspecific is justified.Dedicated to Professor Theodor Günther on the occasion of his 60th birthday     

Al and Fe Biogeochemistry in a floodplain forest: Implications for P retention

We examined spatial and temporal variationsin soil chemistry in a floodplain forest landscape todetermine the effects of flooding on aluminum (Al) andiron (Fe) oxide biogeochemistry and inorganicphosphorus (P_i) sorption capacity. Whenpreviously sorbed P_i was considered, the sorptioncapacities of floodplain and adjacent upland soilswere comparable, suggesting that floodplain soilsrepresent a second line of defense protectingdownstream aquatic ecosystems from agriculturalrun-off. P_i sorption capacity was highlycorrelated with oxalate-extractable Al (Al_o)(r_s = 0.78); Al_o and percent organic matter(OM) were also highly correlated (r_s = 0.72),suggesting the importance of OM-Al complexes in thesesoils. The correlation of oxalate-extractable Fe(Fe_o) with OM (r_s = 0.64) was improved(r_s = 0.80) by removing lower elevation (swale)soils, suggesting that flooding inhibits theassociation of Fe_o with OM. Fe oxidecrystallinity decreased during seasonal flooding, buttotal extractable Fe did not change significantly. Fesolubilized during flooding was either replaced bysediment deposition (252 ± 3 mmol kg^–1yr^–1), and/or reprecipitated locally. Al oxidecrystallinity also decreased during flooding due to asignificant decline in NaOH-extractable Al (Al_N). Al_N concentrations subsequently returned topre-flooding levels, but sediment Al inputs (57 ±3 mmol kg^–1 yr^–1), were insufficient to account for this recovery. Observed Fetransformations suggest the importance offlooding-induced declines in soil redox potential toFe biogeochemistry; observed Al transformationssuggest the importance of complexation reactions withsoil OM to Al biogeochemistry in this floodplainforest.     

Nutrient (N,P, Si) fluxes between marine sediments and water column in coastal and open Adriatic

Nutrient benthic fluxes, as well as sediment phosphorus concentration at the open sea and coastal water stations of the Central and South Adriatic were studied during 1997–98. The fluxes were in the ranges: 0.16–2.67 mmol m^–2 d^–1 (silicate); –0.031–0.164 mmol m^–2 d^–1 (phosphate); –0.51–2.03 mmol m^–2 d^–1 (ammonia); and –1.32–1.62 mmol m^–2 d^–1 (nitrate + nitrite). Silicate flux showed a gradient from the coastal area to the open sea. Ammonia was the main nitrogen species in the flux at the estuary and bay stations, while the sum of nitrate and nitrite was predominant at the open sea stations. Relationships between phosphate and ammonia fluxes (r = 0.699, p<0.01) as well as phosphate and silicate (r = 0.529, p<0.01) were established.     

Light emission from the Fe2+–EDTA–ascorbic acid–H2O2 system strongly enhanced by plant phenolic acids

Oxidative reactions can result in the formation of electronically excited species that undergo radiative decay depending on electronic transition from the excited state to the ground state with subsequent ultra‐weak photon emission (UPE). We investigated the UPE from the Fe²⁺–EDTA (ethylenediaminetetraacetic acid)–AA (ascorbic acid)–H₂O₂ (hydrogen peroxide) system with a multitube luminometer (Peltier‐cooled photon counter, spectral range 380–630 nm). The UPE, of 92.6 μmol/L Fe²⁺, 185.2 μmol/L EDTA, 472 μmol/L AA, 2.6 mmol/L H₂O₂, reached 1217 ± 118 relative light units during 2 min measurement and was about two times higher (P < 0.001) than the UPE of incomplete systems (Fe²⁺–AA–H₂O₂, Fe²⁺–EDTA–H₂O₂, AA–H₂O₂) and medium alone. Substitution of Fe²⁺ with Cr²⁺, Co²⁺, Mn²⁺ or Cu²⁺ as well as of EDTA with EGTA (ethylene glycol‐bis(β‐aminoethyl ether)‐N,N,N′,N′‐tetraacetic acid) or citrate powerfully inhibited UPE. Experiments with scavengers of reactive oxygen species (dimethyl sulfoxide, mannitol, sodium azide, superoxide dismutase) revealed the dependence of UPE only on hydroxyl radicals. Dimethyl sulfoxide at the concentration of 0.74 mmol/L inhibited UPE by 79 ± 4%. Plant phenolics (ferulic, chlorogenic and caffec acids) at the concentration of 870 μmol/L strongly enhanced UPE by 5‐, 13.9‐ and 46.8‐times (P < 0.001), respectively. It is suggested that augmentation of UPE from Fe²⁺–EDTA–AA–H₂O₂ system can be applied for detection of these phytochemicals.     

Oxygen consumption and ammonia accumulation in the hypolimnion of Walker Lake, Nevada

Walker Lake (area = 140 km², Z _mean = 19.3 m) is a large, terminal lake in western Nevada. As a result of anthropogenic desiccation, the lake has decreased in volume by 75% since the 1880s. The hypolimnion of the lake, now too small to meet the oxygen demand exerted by decaying matter, rapidly goes anoxic after thermal stratification. Field and laboratory studies were conducted to examine the feasibility of using oxygenation to avoid hypolimnetic anoxia and subsequent accumulation of ammonia in the hypolimnion, and to estimate the required DO capacity of an oxygenation system for the lake. The accumulation of inorganic nitrogen in water overlaying sediment was measured in laboratory chambers under various DO levels. Rates of ammonia accumulation ranged from 16.8 to 23.5 mg-N m^–2 d^–1 in chambers with 0, 2.5 and 4.8 mg L^–1 DO, and ammonia release was not significantly different between treatments. Beggiatoa sp. on the sediment surface of the moderately aerated chambers (2.5 and 4.8 mg L^–1 DO) indicated that oxygen penetration into sediment was minimal. In contrast, ammonia accumulation was reversed in chambers with 10 mg L^–1 DO, where oxygen penetration into sediment stimulated nitrification and denitrification. Ammonia accumulation in anoxic chambers (18.1 and 20.6 mg-N m^–2 d^–1) was similar to ammonia accumulation in the hypolimnion from July through September of 1998 (16.5 mg-N m^–2 d^–1). Areal hypolimnetic oxygen demand averaged 1.2 g O₂ m^–2 d^–1 for 1994–1996 and 1998. Sediment oxygen demand (SOD) determined in experimental chambers averaged approximately 0.14 g O₂ m^–2 d^–1. Continuous water currents at the sediment-water interface of 5–6 cm s^–1 resulted in a substantial increase in SOD (0.38 g O₂ m^–2 d^–1). The recommended oxygen delivery capacity of an oxygenation system, taking into account increased SOD due to mixing in the hypolimnion after system start-up, is 215 Mg d^–1. Experimental results suggest that the system should maintain high levels of DO at the sediment-water interface (10 mg L^–1) to insure adequate oxygen penetration into the sediments, and a subsequent inhibition of ammonia accumulation in the hypolimnion of the lake.     

Purification and characterization of an<Emphasis Type="Italic"> N</Emphasis>-acetylglucosaminidase produced by a<Emphasis Type="Italic"> Trichoderma harzianum</Emphasis> strain which controls<Emphasis Type="Italic"> Crinipellis perniciosa</Emphasis>

Isolate 1051 of Trichoderma harzianum, a mycoparasitic fungus, was found to impair development of the phytopathogen, Crinipellis perniciosa, in the field. This Trichoderma strain growing in liquid medium containing chitin produced substantial amounts of chitinases. The N-acetylglucosaminidase present in the culture-supernatant was purified to homogeneity by gel filtration and hydrophobic interaction chromatography, as demonstrated by SDS-PAGE analysis. The enzyme had a molecular mass of 36 kDa and hydrolyzed the synthetic substrate -nitrophenyl-N-acetylglucosaminide (NGlcNAc) with Michaelis–Menten kinetics. Maximal activities were determined at pH 4.0 and a temperature range of 50–60°C. K _m and V _max values for NGlcNAc hydrolysis were 8.06 moles ml^–1 and 3.36 moles ml^–1 min^–1, respectively, at pH 6.0 and 37°C. The enzyme was very sensitive to Fe³⁺, Mn²⁺ and Co²⁺ ions, but less sensitive to Zn²⁺, Al³⁺, Cu²⁺ and Ca²⁺. Glucose at a final concentration of 1 mM inhibited 65% of the original activity of the purified enzyme. Determination of the product (reducing sugar) of hydrolysis of C. perniciosa mycelium and scanning electron microscopic analysis revealed that the N-acetylglucosaminidase hydrolyses the C. perniciosa cell wall.     

Binding of phosphate in sediment accumulation areas of the eastern Gulf of Finland,Baltic Sea

The relationships between P and components binding P were studied by analysing the concentrations of N, P, Fe, Mn, Ca and Al in sediments and pore water along the estuarine transect of the River Neva in August 1995. The high sediment organic matter concentration resulted in low surface redox potential and high pore-water o-P concentration, whereas the abundance of amphipods resulted in high surface redox potentials and low pore-water o-P concentration. However, despite the variation in sediment organic matter and the abundance of amphipods, very reduced conditions and slightly variable concentrations of Tot-P (0.7–1.1 mg g^–1 DW) were observed in the 10–15 cm sediment depth along the estuarine gradient, indicating that the pools of mobile P were largely depleted within the depth of 0–15 cm. Multiple regression analysis demonstrated that organic matter and Tot-Fe concentration of the sediment were closely related to the variation in Tot-P concentration of the sediments (r ² = 0.817, n=32). In addition, the high total Fe:P ratio suggested that there is enough Fe to bind P in sediments along the estuarine gradient. However, low Fe_diss concentrations in the pore water of reduced sediment (redox-potential <–50 mV) indicated efficient precipitation of FeS (FeS and FeS₂), incapable to efficiently bind P. Consequently, the low Fe_diss:o-P ratio (< 1) recorded in pore water in late summer implied that Fe³⁺ oxides formed by diffusing Fe_diss in the oxic zone of the sediments were insufficient to bind the diffusing o-P completely. The measured high o-P concentrations in the near-bottom water are consistent with this conclusion. However, there was enough Fe_diss in pore water to form Fe³⁺ oxides to bind upwards diffusing P in the oxic sediment layer of the innermost Neva estuary and the areas bioturbated by abundant amphipods.     

Paleolimnology of Qilu Hu,Yunnan Province,China

Qilu Hu is a large (A = 36.9 km²), shallow (z_max = 6.8 m) lake that lies at an elevation of 1797 m above msl on the Yunnan Plateau, southern China. Lake waters are hard (Mg = 3.2m eq L^–1, Ca = 1.3 meq L^–1 ), fresh (conductivity = 380 S cm ^–1), and productive (Secchi < 40 cm). An 11-m sediment core has a basal ¹⁴C age of 30960 ± 860 B.P. Sediments between 11 m and 6 m are high in % dry weight, rich in clay components Al₂O₃, Fe₂O₃, K₂O, MgO, and low in organic C (6.1%), carbonate-C (<1.0%), total N (<3.2 mg g^–1), and total S (<-1.7 mg g^–1). Diatoms and pollen indicate open-water conditions between 9.0 m and 6.0 m (1342011790 B.P.). Above 6.0 m, CaCO₃ and organic matter concentrations increase relative to clastics. The transition marks a change to shallow-water conditions as inferred from diatoms and pollen, and probably reflects a shift to drier climate. Uppermost (80-0 cm) red clays were deposited rapidly, probably as a consequence of recent (decades to centuries) riparian disturbances (e.g. agriculture, lake-bottom reclamation, urban development). Dates assigned to events in the Qilu Hu profile are tentative because of potential hard-water-lake error.     

Activities of benthic nitrifiers in streams and their role in oxygen consumption

The in situ rates of oxygen consumption by benthic nitrifiers were estimated at 11 study sites in 4 streams. Two methods were used: an in situ respiration chamber method and a method involving conversion of nitrifying potential measurements to in situ rates. Estimates of benthic nitrogenous oxygen consumption (BNOC) rate ranged from 0–380 mmol of O₂ m^–2·day^–1, and BNOC contributed between 0–85% of the total benthic oxygen consumption rate. The activity of nitrifiers residing in the sediments was influenced by O₂ availability, temperature, pH, and substrate. Depending upon site, nitrification could approximate either first-order or zero-order kinetics with respect to ammonium concentration. The source of ammonium for benthic nitrifiers could be either totally from within the sediment or totally from the overlying water. Nitrate produced in the sediments could flux to the water above or be lost within the sediment. The sediments could act as a source (positive flux) or sink (negative flux) for both ammonium (–185 mmol·m^–2·day^–1 to +195 mmol·m^–2·day^–1) and nitrate (–135 mmol·m^–2·day^–1 to +185 mmol·m^–2·day^–1).This study provides evidence to suggest that measurements of down-stream mass flow changes in inorganic nitrogen forms may give poor estimates of in situ rates of nitrification in flowing waters.     

Nitrogen cycling in lake sediments bioturbated by Chironomus plumosus larvae, under different degrees of oxygenation

Sediment cores containing different densities of Chironomus plumosus, ranging from 0 to 12 000 ind. m^–2, were incubated in the laboratory, with 100 and 39% O₂ saturation in the overlying water. Rates of O₂ uptake, and fluxes of the various inorganic N species were measured after addition of ¹⁵NO _inf3 ^su– to the overlying water. The animals enhanced O₂ and NO _inf3 ^su– uptake, due to irrigation. Denitrification of NO _inf3 ^su– coming from the overlying water (Dw) and dissimilatory NO _inf3 ^su– reduction to NH _inf4 ^sup+ (DNRA) represented 20–30 and 4–10% of the NO _inf3 ^su– uptake, respectively. Only 20–40% of the measured NH _inf4 ^sup+ effluxes corresponded to DNRA, the rest was probably due to animal excretion. Nitrite production, mostly from dissimilatory NO _inf3 ^sup– reduction, was detected at both 39 and 100% oxygen saturation. Higher rates of NO _inf2 ^su– production at the lower oxygen concentrations, were probably due to a thinner oxic layer, compared to fully oxygenated waters. The presence of Chironomus plumosus increased nitrification rates, relative to non-inhabited microcosms. However, nitrification rates were low compared to Dw, probably due to low numbers of nitrifiers in the sediment. At 39% oxygen saturation, rates of nitrification and denitrification of NO _inf3 ^su– generated within the sediment were not measurable.     

Chemolithoautotrophy in the marine,magnetotactic bacterial strains MV-1 and MV-2

Magnetite-producing magnetotactic bacteria collected from the oxic–anoxic transition zone of chemically stratified marine environments characterized by O₂/H₂S inverse double gradients, contained internal S-rich inclusions resembling elemental S globules, suggesting they oxidize reduced S compounds that could support autotrophy. Two strains of marine magnetotactic bacteria, MV-1 and MV-2, isolated from such sites grew in O₂-gradient media with H₂S or thiosulfate (S₂O₃^2–) as electron sources and O₂ as electron acceptor or anaerobically with S₂O₃^2– and N₂O as electron acceptor, with bicarbonate (HCO₃^–)/CO₂ as sole C source. Cells grown with H₂S contained S-rich inclusions. Cells oxidized S₂O₃^2– to sulfate (SO₄^2–). Both strains grew microaerobically with formate. Neither grew microaerobically with tetrathionate (S₄O₆^2–), methanol, or Fe²⁺ as FeS, or siderite (FeCO₃). Growth with S₂O₃^2– and radiolabeled ¹⁴C-HCO₃^– showed that cell C was derived from HCO₃^–/CO₂. Cell-free extracts showed ribulose 1,5-bisphosphate carboxylase/oxygenase (RubisCO) activity. Southern blot analyses indicated the presence of a form II RubisCO (cbbM) but no form I (cbbL) in both strains. cbbM and cbbQ, a putative post-translational activator of RubisCO, were identified in MV-1. MV-1 and MV-2 are thus chemolithoautotrophs that use the Calvin–Benson–Bassham pathway. cbbM was also identified in Magnetospirillum magnetotacticum. Thus, magnetotactic bacteria at the oxic–anoxic transition zone of chemically stratified aquatic environments are important in C cycling and primary productivity.     

Buffered,phosphate-containing media suitable for aluminum toxicity studies

Studies of Al rhizotoxicity sometimes require the use of well-defined rooting media. For that reason, buffers and phosphate are often omitted from Al solutions for which species composition must be determined precisely. Homopipes and succinate appear to be suitable buffers for short-term studies with seedlings of an Al-sensitive wheat (Triticum aestivum L. cv. Scout 66) and white clover (Trifoliau repens L. cv. Huia). In the case of homopipes (homopiperazine-N,N-bis-2-[ethane-sulfonic acid]), a slight inhibition of root elongation must be taken into account, but no binding of Al³⁺ was observed. In the case of succinate, no inhibition of root elongation was observed, but Al³⁺ binding must be considered. Phosphate-containing media remain free of solid-phase or polynuclear species whenever {Al³⁺}²{HPO₄ ^2-}{OH^–}³ < 10^–47.0 (or {Al³⁺}{HPO₄ ^2-}{OH^–}< 10^–22.7) and when {Al³⁺}³ / {H⁺}³ < 10^8.8. These ion activity products, that define stable Al solutions in the laboratory, appear to apply in soils also, according to an analysis of published data. The published equilibrium values {AlH₂PO₄ ²⁺} / ({Al³⁺}{H₂PO₄ ^–}) = 10^3.0, {AlHPO₄ ⁺} / ({Al³⁺}{HPO₄ ^2-}) = 10^7.0, and {Alsuccinate⁺} / ({Al³⁺}{succinate ^2-}) = 10^4.62 appear to be suitable, because solution toxicity could be accounted for entirely on the basis of computed Al³⁺ even in solutions containing high levels of Alsuccinate⁺ and AlHPO₄ ⁺ (in every case {AlHPO₄ ⁺}>> {AlH₂PO₄ ²⁺}). Thus, AlHPO₄ ⁺ and Alsuccinate⁺ were not toxic at achieved concentrations.     

Seasonal variation in phytoplankton primary production in relation to light and nutrients in Lake Awasa,Ethiopia

The biomass and primary production of phytoplankton in Lake Awasa, Ethiopia was measured over a 14 month period, November 1983 to March 1985. The lake had a mean phytoplankton biomass of 34 mg chl a m^–3 (n = 14). The seasonal variation in phytoplankton biomass of the euphotic zone (mg chl a m^–2 h^–1) was muted with a CV (standard deviation/mean) of 31%. The vertical distribution of photosynthetic activity was of a typical pattern for phytoplankton with light inhibition on all but overcast days. The maximum specific rates of photosynthesis or photosynthetic capacity (Ø_max) for the lake approached 19 mg O₂ (mg chl a)^–1 h^–1, with high values during periods of low phytoplankton biomass. Areal rates of photosynthesis ranged between 0.30 to 0.73 g O₂ m^–2 h^–1 and 3.3 to 7.8 g O₂ m^–2 d^–1. The efficiency of utilisation of PhAR incident on the lake surface varied from 2.4 to 4.1 mmol E^–1 with the highest efficiency observed corresponding to the lowest surface radiation. Calculated on a caloric basis, the efficiency ranged between 1.7 and 2.9%. The temporal pattern of primary production by phytoplankton showed limited variability (CV = 21 %).     

Nitrification, denitrification, and nitrate ammonification in sediments of two coastal lagoons in Southern France

Summary Seasonal and diurnal variations in sediment-water fluxes of O₂, NO ₃ ^– , and NH ₄ ⁺ as well as rates of nitrification, denitrification, and nitrate ammonification were determined in two different coastal lagoons of southern France: The seagrass (Zostera noltii) dominated tidal Bassin d'Arcachon and the dystrophic Etang du Prévost. Overall, denitrification rates in both Bassin d'Arcachon (<0.4 mmol m^–2 d^–1) and Etang du Prévost (<1 mmol m^–2 d^–1) were low. This was mainly caused by a combination of low NO ₃ ^– concentrations in the water column and a low nitrification activity within the sediment. In both Bassin d'Arcachon and Etang du Prévost, rates of nitrate ammonification were quantitatively as important as denitrification.Denitrification played a minor role as a nitrogen sink in both systems. In the tidal influenced Bassin d'Arcachon, Z. noltii was quantitatively more important than denitrification as a nitrogen sink due to the high assimilation rates of the plants. Throughout the year, Z. noltii stabilized the mudflats of the bay by its well- developed root matrix and controlled the nitrogen cycle due to its high uptake rates. In contrast, the lack of rooted macrophytes, and dominance of floating macroalgae, made nitrogen cycling in Etang du Prévost more unstable and unpredictable. Inhibition of nitrification and denitrification during the dystrophic crisis in the summer time increased the inorganic nitrogen flux from the sediment to the water column and thus increased the degree of benthic-pelagic coupling within this bay. During winter, however, benthic microalgae colonizing the sediment surface changed the sediment in the lagoon from being a nitrogen source to the over lying water to being a sink due to their high assimilation rates. It is likely, however, that this assimilated nitrogen is liberated to the water column at the onset of summer thereby fueling the extensive growth of the floating macroalgae, Ulva sp. The combination of a high nitrogen coupling between sediment and water column, little water exchange and low denitrification rates resulted in an unstable system with fast growing algal species such as phytoplankton and floating algae.     

A comparison of oxygen,nitrate, and sulfate respiration in coastal marine sediments

Aerobic respiration with oxygen and anaerobic respiration with nitrate (denitrification) and sulfate (sulfate reduction) were measured during winter and summer in two coastal marine sediments (Denmark). Both aerobic respiration and denitrification took place in the oxidized surface layer, whereas sulfate reduction was most significant in the deeper, reduced sediment. The low availability of nitrate apparently limited the activity of denitrification during summer to less than 0.2 mmoles NO ₃ ^– m^–2 day^–1, whereas activities of 1.0–3.0 mmoles NO ₃ ^– m^–2 day^–1 were measured during winter. Sulfate reduction, on the contrary, increased from 2.6–7.6 mmoles SO ₄ ^2– m^–2 day^–1 during winter to 9.8–15.1 mmoles SO ₄ ^2– m^–2 day^–1 during summer. The aerobic respiration was high during summer, 135–140 mmoles O₂ m^–2 day^–1, as compared to estimated winter activities of about 30 mmoles O₂ m^–2 day^–1. The little importance of denitrification relative to aerobic respiration and sulfate reduction is discussed in relation to the availability and distribution of oxygen, nitrate, and sulfate in the sediments and to the detritus mineralization.     

Measurement of Nitrous Oxide Reductase Activity in Aquatic Sediments

Denitrification in aquatic sediments was measured by an N₂O reductase assay. Sediments consumed small added quantities of N₂O over short periods (a few hours). In experiments with sediment slurries, N₂O reductase activity was inhibited by O₂, C₂H₂, heat treatment, and by high levels of nitrate (1 mM) or sulfide (10 mM). However, ambient levels of nitrate (<100 μM) did not influence activity, and moderate levels (about 150 μM) induced only a short lag before reductase activity began. Moderate levels of sulfide (<1 mM) had no effect on N₂O reductase activity. Nitrous oxide reductase displayed Michaelis-Menten kinetics in sediments from freshwater (K_m = 2.17 μM), estuarine (K_m = 14.5 μM), and alkaline-saline (K_m = 501 μM) environments. An in situ assay was devised in which a solution of N₂O was injected into sealed glass cores containing intact sediment. Two estimates of net rates of denitrification in San Francisco Bay under approximated in situ conditions were 0.009 and 0.041 mmol of N₂O per m² per h. Addition of chlorate to inhibit denitrification in these intact-core experiments (to estimate gross rates of N₂O consumption) resulted in approximately a 14% upward revision of estimates of net rates. These results were comparable to an in situ estimate of 0.022 mmol of N₂O per m² per h made with the acetylene block assay.     

Ulva rigida (Ulvales,Chlorophyta) tank culture as biofilters for dissolved inorganic nitrogen from fishpond effluents

Ulva rigida was cultivated in 7501 tanks at different densities with direct and continuous inflow (at 2, 4, 8 and 12 volumes d^–1) of the effluents from a commercial marine fishpond (40 metric tonnes, Tm, of Sparus aurata, water exchange rate of 16 m³ Tm^–1) in order to assess the maximum and optimum dissolved inorganic nitrogen (DIN) uptake rate and the annual stability of the Ulva tank biofiltering system. Maximum yields (40 g DW m^–2 d^–1) were obtained at a density of 2.5 g FW 1^–1 and at a DIN inflow rate of 1.7 g DIN m^–2 d^–1. Maximum DIN uptake rates were obtained during summer (2.2 g DIN M^–2 d^–1), and minimum in winter (1.1 g DIN m^–2 d^–1) with a yearly average DIN uptake rate of 1.77 g DIN m^–2 d^–1 At yearly average DIN removal efficiency (2.0 g DIN m^–2 d^–1, if winter period is excluded), 153 m² of Ulva tank surface would be needed to recover 100% of the DIN produced by 1 Tm of fish.Abbreviations DIN= dissolved inorganic nitrogen (NH _inf4 ^sup+ + NO _inf3 ^sup– + NO _inf2 ^sup– ); - FW= fresh weight; - DW= dry weight; - PFD= photon flux density; - V= DIN uptake rate     

Uptake of iron byGeotrichum candidum,a non-siderophore producer

Summary Geotrichum candidum (isolate 1–9) pathogenic on citrus fruits, appears to lack siderophore production. Iron uptake byG. candidum is mediated by two distinct iron-regulated, energy-and temperature-dependent transport systems that require sulfhydryl groups. One system exhibits specificity for either ferric or ferrous iron, whereas the other exhibits specificity for ferrioxamine-B-mediated iron uptake and presumably other hydroxamate siderophores. Radioactive iron uptake from⁵⁹FeCl₃ showed an optimum at pH 6 and 35° C, and Michaelis-Menten kinetics (apparentK _m = 3 m,V _max = 0.054 nmol · mg^–1 · min^–1). The maximal rate of Fe²⁺ uptake was higher than Fe³⁺ (V _max = 0.25 nmol · mg^–1 · min^–1) but theK _m was identical. Reduction of ferric to ferrous iron prior to transport could not be detected. The ferrioxamine B system exhibits an optimum at pH 6 and 40° C and saturation kinetics (K _m = 2 M,V _max = 0.22 nmol · mg^–1 · min^–1). The two systems were distinguished as two separate entities by negative reciprocal competition, and on the basis of differential response to temperature and phenazine methosulfate. Mössbauer studies revealed that cells fed with either⁵⁷FeCl₃ or⁵⁷FeCl₂ accumulated unknown ferric and ferrous binding metabolites.     

Physical and structural characterization of yersiniophore, a siderophore produced by clinical isolates of Yersinia enterocolitica

Clinical isolates of Yersinia enterocolitca, which belong to mouse-lethal serotypes, produce the siderophore yersiniophore. Siderophore production was shown to be iron regulated and to reach maximum production in late log phase. Yersiniophore is a fluorescent siderophore with maximum excitation at 270 nm and a major emission peak at 428 nm. Absorption maxima were seen at 210 and 250 nm with a low broad peak from 280 to 320 nm. Purification of unchelated yersiniophore for structural analysis was made difficult by low yields (1–2 mg mg^-1), and susceptibility to acid hydrolysis, oxidation and possibly polymerization. Yersinophore was therefore purified as an Al³⁺ chelate, which was found to be stable in solution for several weeks. To purify Al³⁺-yersiniophore, unchelated yersiniophore was first extracted from culture supernatants with dichloromethane, concentrated by rotary evaporation and adsorbed to a DEAE-sephacel column. Al³⁺-yersiniophore was eluted with 0.01 m AlCl₃ and further purified by HPLC. The structure was established by a combination of elemental analysis, high resolution mass spectrometry and two-dimensional NMR experiments. Yersiniophore is a phenolate-thiazole siderophore with the formula C₂₁H₂₄N₃O₄S₃Al and a molecular weight of 505.07404 when chelated to Al³⁺. The structure of yersiniophore was determined to be closely related to the structures of pyochelin, produced by Pseudomonas aeruginosa, and anguibactin, produced by Vibrio anguillarum.     

Inactivation of the plasma membrane ATPase ofSchizosaccharomyces pombe by hydrogen peroxide and by the fenton reagent (Fe2+/H2O2): Nonradicalvs. Radical-induced oxidation

In the absence of added Fe²⁺, the ATPase activity of isolatedSchizosaccharomyces pombe plasma membranes (5–7 μmolP _i per mg protein per min) is moderately inhibited by H₂O₂ in a concentration-dependent manner. Sizable inactivation occurs only at 50–80 mmol/L H₂O₂. The process, probably a direct oxidative action of H₂O₂ on the enzyme, is not induced by the indigenous membrane-bound iron (19.3 nmol/mg membrane protein), is not affected by the radical scavengers mannitol and Tris, and involves a decrease of both theK _m of the enzyme for ATP and theV of ATP splitting. On exposing the membranes to the Fenton reagent (50 μmol/L Fe²⁺ +20 mmol/L H₂O₂), which causes a fast production of HO⁻ radicals, the ATPase is 50–60% inactivated and 90% of added Fe²⁺ is oxidized to Fe³⁺ within 1 min. The inactivation occurs only when Fe²⁺ is added before H₂O₂ and can thus bind to the membranes. The lack of effect of radical scavengers (mannitol, Tris) indicates that HO⁻ radicals produced in the bulk phase play no role in inactivation. Blockage of the inactivation by the iron chelator deferrioxamine implies that the process requires the presence of Fe²⁺ ions bound to binding sites on the enzyme molecules. Added catalase, which competes with Fe²⁺ for H₂O₂, slows down the inactivation but in some cases increases its total extent, probably due to the formation of the superoxide radical that gives rise to delayed HO⁻ production.