Ecological status of some Finnish rivers evaluated using benthic diatom communities

The applicability of some diatom methods to quality assessment of differenttypes of Finnish rivers was compared using dominant taxa of diatomcommunities and nutrient level estimations using Trophic Diatom Index(TDI) and a new Phosphorus Diatom Equation (PDE) [Tot. P_calc (g l^-1) = 39 – 0.278 ol-% – 0.117 olme-% – 0.346me-% – 0.006 meeu-% + 1.193 eu-%; n = 97; r² = 0.742;SE = 17.4]. The overall quality estimations were done by Index ofPollution Sensitivity (IPS) and by Generic Diatom Index (GDI); limit valuesfor quality classes are proposed.     

Diatom–environmental relationships and nutrient transfer functions from contrasting shallow and deep limestone lakes in Ontario, Canada

Diatom assemblages were analysed in the surface sediments of 44 alkaline lakes in south-western Ontario, Canada, and combined with a pre-existing 58 south-eastern Ontario lake set: (1) to determine if shallow, polymictic Ontario lakes contain different diatom assemblages from deeper, dimictic lakes, and if so, which environmental variables most influence assemblages; (2) to improve the existing transfer functions; (3) to construct and compare transfer functions separately for dimictic, deep lakes and for polymictic, shallow lakes. Polymictic and dimictic lakes covered a similar nutrient range (spring total phosphorus (TP)=4–54 g/l, spring total nitrogen (TN)=200–927 g/l; n=101) and spring pH levels (7.6–9; n=101). However, polymictic lakes were shallower (median mean depth = 2.9 m vs. 7.3 m in dimictic lakes). Benthic diatoms (average 60% relative abundance) dominated the polymictic lakes, whereas planktonic diatoms (average 60%) dominated dimictic lakes. A Canonical Correspondence Analyses with forward selection (p < 0.05, 999 Monte Carlo permutations) identified TP, alkalinity, watershed to volume ratios and lake depth as the most important measured environmental variables influencing diatom distribution in both polymictic and dimictic lakes. Additionally, pH was identified as an important variable in polymictic lakes, whereas TN was also forward selected in the dimictic lakes. Adding more lakes to the original southern Ontario calibration set improved the TN transfer function (r² _jack=0.42, root mean squared error of prediction (RMSEP)_jack=0.11 [log g TN/l]), although there was a high systematic error in the revised model (r² _residual = 0.48). However, the strongest TP model was derived from the polymictic lakes (r² _boot =0.44, RMSEP_boot=0.20 [log g TP/l]), which was the smallest lake set (n=30) with the lowest number of diatom species. The stronger TP model from the polymictic lakes may be partly due to the relatively low macrophyte cover in our polymictic lakes, which may lead to stronger benthic–pelagic coupling than in lakes with large macrophyte populations. Additionally, our study suggests that the Chrysophyceae cyst:diatom frustule ratio may be useful for indicating trends in TP levels of 35 g/l in alkaline lakes that are dimictic, but is not necessarily indicative of trophic state changes in shallow, polymictic lakes. Our study demonstrates that it may be important to construct separate diatom-based nutrient transfer functions for polymictic and dimictic lakes.     

Correlation of surface sediment diatoms with the present lake water pH in 28 Algoma lakes,Ontario, Canada

The surface sediment diatom analysis of 28 Algoma lakes (pH 4.40–8.13) indicates that even though each lake has a widely different aquatic environment and characteristic diatom assemblage, a definite relationship exists between the lake water pH and their diatom assemblages. In the acidic lakes acidobiontic and acidophilous diatom species predominate whereas in circumneutral and alkaline lakes circumneutral and alkaliphilous diatoms were most common. Cluster analysis of the pH indicator diatom assemblages grouped the study lakes into three distinct cluster groups. These groups also closely corresponded to lake water pH. On the basis of published ecological information as well as their presence in our study lakes, the pH indicator status of a number of diatom taxa have been discussed. A detailed listing of the diatom taxa identified and their pH indicator status is provided in order to facilitate their use in future diatom-inferred pH studies.     

An investigation of the effects of external acidification on sodium transport,internal pH and membrane potential in barnacle muscle fibers

Summary Radiosodium efflux from barnacle muscle fibers is a function of pH _e , the threshold pH _e for stimulation of Na efflux into HCO ₃ ^– -artificial sea water (ASW) being 6.8 and the fixed thresholdpCO₂ (in an open CO₂ system) being approximately 30 mm Hg. Acidification of ASW containing non-HCO ₃ ^– buffer is without effect on the Na efflux. The Na efflux following stimulation by reducing the pH of 10mM HCO ₃ ^– -ASW from 7.8 to 6.3 is reduced by 17.3% as the result of microinjecting 100mM EGTA, and increased by 32.6% as the result of microinjecting 0.5M ATP. The Na efflux into K-free HCO ₃ ^– -ASW is markedly stimulated by external acidification. Ouabain-poisoned fibers are more responsive to a low pH _e than unpoisoned fibers. Applying the 2-¹⁴C-DMO technique, it is found that fibers bathed in 10mM HCO ₃ ^– -ASW at pH 7.8 have an internal pH of 7.09±0.106 (mean±SD), whereas fibers bathed in 25mM TRIS-ASW at pH 7.8 have a pH _i of 7.28±0.112. The relationship between pH _i and pH _e as external pH is varied by adding H⁺ is linear. Measurements of the resting membrane potential indicate that external acidification in the presence of HCO ₃ ^– as buffer is accompanied by a fall inE _m , the threshold pH _e being 7.3 both at 24 and 0°C. This sensitivity amounts to 8.2 mV per pH unit (at 24°C) over a wide range of pH _e . Membrane resistance following external acidification remains unchanged. Microinjection of the protein inhibitor of Walsh before external acidification fails to stop depolarization from occurring. Cooling to 0°C also fails to abolish depolarization following acidification. Whereas external ouabain and ethacrynic acid reduceE _m in the absence or presence of acidification, DPH hyperpolarizes the membrane or arrests depolarization both at 24 and 0°C. This effect of DPH at 0°C is seen in the absence or presence of acidification. It is suggested that depolarization following acidification of a HCO ₃ ^– -containing medium is due to activation of a Cl^–-and/or HCO ₃ ^– -pump and that ouabain and ethacrynic acid reducesE _m by abolishing uncoupled Na transport.     

Development of diatom-based salinity models for paleoclimatic research from lakes in British Columbia (Canada)

Diatoms were identified and enumerated from the surface sediments of 65 lakes located on the Cariboo and Chilcotin Plateaux (British Columbia, Canada). These lakes span a large gradient in lakewater ionic concentration (fresh through hypersaline) and composition, as well as other physical/chemical variables. Almost all of the study lakes had higher salinities in the late-summer than in the spring. The lakes with spring salinities >8 g l^–1 showed the largest seasonal increases in salinity. Ionic composition was similar in the spring and late-summer for most lakes. Both ionic concentration (i.e. salinity) and composition were important environmental variables that could account for the different diatom floras in the lakes. Diatom assemblages characteristic of carbonate-dominated and sulfate-dominated waters were identified. Other variables such as water depth and phosphorus concentration were also important.The majority (87%) of diatom taxa had estimated salinity optima < 3 g l^–1 Halophilic diatom taxa had broader tolerances to salinity when compared to the fresh water taxa, however taxa with narrow and broad tolerances could be identified across the salinity gradient. Species diversity was weakly but significantly correlated to lakewater salinity (r ² = 0.18 to 0.3, P < 0.05).Salinity inference models were developed based on the relationship between the diatom assemblages and the spring, late-summer and average salinity. The correlations between the measured and diatominferred salinity, based on the spring (r = 0.95), late-summer (r = 0.94) and average (r = 0.95) salinity data, are high because there was an extremely strong correlation (r = 0.98) between the log transformed spring and late-summer measured salinities. These salinity reconstruction models provide a tool that can be used to infer past climatic changes as part of paleolimnological studies from appropriate closed-basin lakes in British Columbia.     

Ion fluxes and pH changes induced by trans-plasmalemma electron transfer and fusicoccin inLemna gibba L. (strain G1)

During the reduction of extracellular [Fe(CN)₆]^3– at the plasmalemma of intact, K⁺-starvedLemna gibba L. fronds, the external medium was acidified and K⁺ released, in the absence of inhibitors with rates of 10 e^–/8.5 H⁺/1.5 K⁺ (mol·(g FW)^–1·^–1). In K⁺ plants the larger K⁺ efflux caused a lag phase in extracellular acidification and a change in rates to 10 e^–/6 H⁺/4 K⁺ and in the presence of CN^–+salicylhydroxamic acid at pH 5 to 5.2 e^–/0 H⁺/6.6 K⁺. The e^– transfer was accompanied by a membrane depolarization of up to 100 mV and a cytosolic acidification of about 0.6 pH units, but only in K⁺ plants, where the extracellular acidification was smaller. These results indicate that a stimulation of the plasmalemma H⁺-ATPase may be triggered either by a cytosolic acidification or by a strong membrane depolarization. It is concluded that the redox system catalyses only uncoupled e^– transfer without H⁺ transfer across the plasmalemma. The obligatory, but secondary charge compensation is partially achieved by the rapid K⁺ release upon membrane depolarization and partially by the activity of the plasma membrane H⁺-ATPase, but not by an e^–/anion exchange. The extracellular acidification during [Fe(CN)₆]^3– reduction is generated by the conversion of a strong trivalent into a strong tetravalent anion. This acidification is caused by changes in the concentration ratio of strong cations to strong anions. Efflux of K⁺ and not the production of organic acids or NAD(P)H oxidation is the chemical cause of the measurable cytosolic acidification. Extracellular acidification was inversely correlated with intracellular acidification. Similarly, fusicoccin-induced pH changes were correlated with changes in the strong-ion concentration difference. Extracellular ± FC-dependent acidification and intracellular alkalinization of up to 0.6 pH units were strongly dependent on K⁺ fluxes. The ferricyanide-triggered trans-plasmalemma electron-transfer system is an example of how measurable pH changes are the consequence and not the cause of charge-transfer-induced changes in strong-ion fluxes.Abbreviations DCCD dicyclohexylcarbodiimide - E_m electrical membrane potential difference - FC fusicoccin - pH_c cytosolic pH - FW fresh weight - PM plasmalemma - SHAM salicylhydroxamic acid - SID strong-ion concentration difference This work was supported by the Deutsche Forschungsgemeinschaft. We gratefully acknowledge the Alexander von Humboldt award donated to J.G. We thank Professor Ulrich Lüttge (TH Darmstadt, FRG) for his kind support and Annett Ehrhardt and Dr. Karl Fischer (TH Darmstadt, FRG) for their valuable help with Cl^– and CO₂ experiments. Special thanks are due to Professor Erasmo Marrè (Università di Milano, Italy) for continuous discussions and also to Professor Alessandro Ballio (Università di Roma, Italy) for their kind gifts of fusicoccin.     

pH equilibration in human erythrocyte suspensions

Summary A stopped-flow rapid reaction apparatus was used to study the rate of pH equilibration in human red cell suspensions. Flux of OH^– or H⁺ was determined over a wide range of extracellular pH (4–11) in CO₂-free erythrocyte suspensions. In these experiments, an erythrocyte suspension at pH 7.3 is rapidly mixed with an equal volume of NaCl solution at 3.0>pH>11.5. The pH of the extracellular fluid of the mixture changes rapidly as OH^– or H⁺ moves across the red cell membrane. Flux and velocity constants can be calculated from the initiald pH/dt using the known initial intra- and extracellular pH. It was found that the further the extracellular pH is from 7.3 (in either direction from 4–11), the greater the absolute value of total OH^– and/or H⁺ flux. Pretreatment with SITS (4-acetamido-4-isothiocyanostilbene-2,2-disulfonic acid), a potent anion exchange inhibitor, greatly reduces flux over the entire pH range, while exposure to valinomycin, a potassium ionophore, has no measurable effect. These data suggest that (i) both H⁺ and OH^– may be moving across the red cell membrane at all pH; (ii) the species dominating pH equilibration is probably dependent on the extracellular pH, which determines the magnitude of the driving gradient for each ion; and (iii) the rapid exchange pathway of the erythrocyte membrane may be utilized for both H⁺ and OH^– transport during CO₂-free pH equilibration.     

pH modulation of the kinetics of rabbit jejunal,brush-border folate transport

Summary In jejunal brush-border membrane vesicles, an outwardly directed OH^– gradient (in>out) stimulates DIDS-sensitive, saturable folate (F) uptake (Schron, C.M. 1985.J. Clin. Invest. 76:2030–2033), suggesting carrier-mediated folate: OH^– exchange (or phenomenologically indistinguishable H⁺: folate cotransport). In the present study, the precise role of pH in the transport process was elucidated by examining F uptake at varying pH. For pH gradients of identical magnitude, F uptake (0.1 M) was greater at lower (pH_int/pH_ext: 5.5/4.5) compared with higher (6.5/5.5) pH ranges. In the absence of a pH gradient, internal Ftrans stimulated DIDS-sensitive³H-folate uptake only at pH6.0. Since stepwise increments ininternal pH (4.57.5; pH_ext=4.5) stimulated F uptake, an inhibitory effect of higherinternal pH was excluded. In contrast, with increasing external pH (4.356.5; pH_int=7.8), a 50-fold decrement in F uptake was observed (H⁺ K _m =12.8±1.2 M). Hill plots of these data suggest involvement of at least one H⁺ (OH^–) at low pH (monovalent F^– predominates) and at least 2 H⁺ (OH^–) at high pH (divalent F^–2 predominates). Since an inside-negative electrical potential did not affect F uptake at either pH_ext 4.55 or 5.8, transport of F^– and F^–2 is electroneutral. Kinetic parameters for F^– and F^–2 were calculated from uptake data at pH_ext 4.55 and 5.0. Comparison of predictedvs. experimentally determined kinetic parameters at pH_ext5.8 (K _m =1.33vs. 1.70 M;V _max=123.8vs. 58.0 pmol/mg prot min) suggest that increasing external pH lowers theV _max, but does not affect theK _m for carrier-mediated F transport. These data are consistent with similarK _i ^' s for sulfasalazine (competitive inhibitor) at pH_ext 5.35 and 5.8 (64.7 and 58.5 M, respectively). In summary, the jejunal F carrier mediates electroneutral transport of mono- and divalent F and is sensitive to external pH with a H⁺ K _m (or OH^– lC₅₀) corresponding to pH 4.89. External pH effects theV _max, but not theK _m for carriermediated F uptake suggesting a reaction mechanism involving a ternary complex between the outward-facing conformation of the carrier and the transported ions (F and either OH^– or H⁺),rather than competitive binding that is mutually exclusive.     

Diatom distribution and diatom inferred pH in the sediment of four alpine lakes

The diatom distribution in the sediment of four alpine lakes in the central alpine area (Ötztaler Alpen, Tyrolia, and Mölltal, Carinthia) has been analysed in order to assess possible influences of atmospheric pollutants on remote lakes. The lakes are located more than 2000 m above sea level on poorly buffered bedrock (granite, gneiss and mica schist) and are therefore susceptible to acidification by acid deposition. The pH history of the lakes has been calculated by using Index B from Renberg & Hellberg (1982), which has been adjusted to central European conditions and applied in this region for the first time. The results indicate that there have been no pH decreases in three of the lakes, while in one lake diatom composition and distribution change significantly towards the sediment surface. But taxonomical problems and difficulties in the pH classification of the rarely found diatoms complicate the interpretation of the results. Important taxa will be discussed in view of their ecological significance.     

Disruption of sulfur cycling and acid neutralization in lakes at low pH

Experimental acidification of a softwater lake to below pH 5 fundamentally changed the sulfur cycle and lowered internal alkalinity generation (IAG). Prior to reaching pH 4.5, the balance of sulfur reduction and oxidation reactions within the lake was in favour of reduction, and the lake was a net sink for sulfate. In the four years at pH 4.5 the balance of reduction and oxidation reactions was in favour of oxidation, and there was a net production of sulfate (SO₄ ^2–) within the lake. Evidence indicating a decrease in net SO₄ ^2– reduction at pH 4.5 was also obtained in an anthropogenically acidified lake that had been acidified for many decades. In both lakes, the decrease in net SO₄ ^2– reduction appeared to be linked not to a simple inhibition of SO₄ ^2– reduction but rather to changes in benthic ecosystem structure, especially the development of metaphytic filamentous green algae, which altered the balance between SO₄ ^2– reduction and sulfur oxidation.At pH's above 4.5, net SO₄ ^2– reduction was the major contributor to IAG in the experimental lake, as it is in many previously studied lakes at pH 5 and above. At pH 4.5, the change in net annual SO₄ ^2– reduction (a decrease of 110%) resulted in a 38% decrease in total IAG. Because of the important role of net SO₄ ^2– reduction in acid neutralization in softwater lakes, models for predicting acidification and recovery of lakes may need to be modified for lakes acidified to pH <5.     

pH Modulation of the kinetics of rabbit jejunal,brush-border folate transport

Summary In jejunal brush-border membrane vesicles, an out-wardly directed OH^– gradient (in>out) stimulates DIDS-sensitive, saturable folate (F) uptake (Schron, C.M., 1985).J. Clin. Invest. 76:2030–2033), suggesting carrier-mediated folate: OH^– exchange (or phenomenologically indistiguishable H⁺: folate cotransport). In the present study, the precise role of pH in the transport process was elucidated by examinin F uptake at varying pH. For pH gradients of identical magnitude, F uptake (0.1 M) was geater at lower (pH_int/pH_ext:5.5/4.5) compared with higher (6.5/5.5) pH ranges. In the absence of a pH gradient, internal Ftrans stimulated DIDS-sensitive³H-folate uptake only at pH6.0. Since setepwise increments ininternal pH (4.57.5; pH_ext=4.5) stimulated F uptake, an inhibitory effect of higherinternal pH was excluded. In contrast, with increasing external pH(4.356.5; pH_int=7.8), a 50-fold decrement in F uptake was observed (H⁺ K _m =12.8±1.2m). Hill plots of these data suggest involvement of at least one H⁺ (OH^–) at high pH (divalent F^–2 predominates). Since an inside-negative electrical potential did not affect F uptake at either pH_ext 4.55 or 5.8, transport of F^– and F^–2 is electroneutral. Kinetic parameters for F^– and F^–2 were calculated from uptake data at pH_ext 4.55 and 5.0. Comparision of predictedvs. experimentally determined kinetic parameters at pH_ext 5.8 (K _m =1.33vs. 1.70 m;V _max=12.8vs. 58.0 pmol/mg prot min) suggest that increasing external pH lowers theV _max, but does not affect thatK _m, for carrier-mediated F transport. These data are consistent with similarK _i's for sulfasalazine (competitive inhibitor) at pH_ext 5.35 and 5.8 (64.7 and 58.5 m, respectively). In summary, the jejunal F carrier mediates electroneutral transport of mono- and divalen F and is sensitive to extermal pH with a H⁺ K _m (or OH^– IC₅₀) corresponding to pH 4.89. External pH affects theV _max, but not theK _m for carriermediated F uptake suggesting a reaction mechanism involving a ternary complex between the outward-facing conformation of the carrier and the transported ions (F and either OH^– or H⁺) rather than competitive binding that is mutually exclusive.     

The intracellular pH of isolated,photosynthetically active Asparagus mesophyll cells

The intracellular pH of isolated, photosynthetically active mesophyll cells of Asparagus sprengeri Regel has been determined, in the light and dark, by the distribution of the weak acid 5,5-dimethyl-[2-¹⁴C]oxazolidine-2,4-dione ([¹⁴C]DMO) between the cells and the liquid medium. [¹⁴C]DMO was taken up rapidly, reaching equilibrium in 7–10 min of incubation, but was not metabolized by the cells, and intracellular binding of the compound was minimal. The intracellular pH, measured at saturating light fluence and 1.5 mM sodium bicarbonate, was found to remain relatively constant at 6.95–7.21 over the external pH range of 5.5–7.2. Illumination of the cells increased the intracellular pH compared to dark controls. The pH of the cytoplasm, excluding and including the chloroplasts (cytoplasmic and bulk cytoplasmic, respectively) was calculated from the experimentally derived intracellular [¹⁴C]DMO concentration and estimates of the vacuolar, chloroplastic and cytoplasmic volumes. The calculated cytoplasmic pH was similar in the light and dark, being 7.75 and 7.74, respectively, while the calculated pH of bulk cytoplasm was 7.85 in the light and 7.49 in the dark. Theoretical analysis indicated that intracellular pH is a good indicator of changes in the bulk cytoplasmic pH but insensitive to changes in vacuolar pH. The external pH optimum for photosynthesis (O₂ evolution) of isolated Asparagus cells was pH 7.2. At pH 8.0 photosynthesis was inhibited by 30% and at pH 5.25 by 45%. Inhibition at alkaline pH may be the result of a decrease in the pH gradient between the cells and the medium, causing CO₂ limitation in the cell. At acid pH, decrease in internal pH caused by substantial accumulation of inorganic carbon may account for the loss in photosynthetic activity.Abbreviations [¹⁴C]DMO 5,5-dimethyl[2-¹⁴C]oxazolidine-2,4-dione - pH_i overall intracellular pH - pH_e pH of external medium     

The relationship between surface sediment diatom assemblages and pH in 33 Galloway lakes: Some regression models for reconstructing pH and their application to sediment cores

The species composition of surface sediment diatom assemblages in 33 Galloway lakes, pH range c. 4.5 to 7.4, is related in a statistically significant manner to water acidity (or to factors closely associated with pH). Predictive models of summer mean pH, using simple regression equations of Index B (Renberg & Hellberg, 1982) with both Scandinavian and Galloway data sets, and multiple regression equations using diatom pH preference groups and individual species, are described and applied to fossil diatoms in six sediment cores. Although multiple regression of individual species gives the highest correlation coefficient (r²=0.87) in the modern data set this method is least appropriate for reconstructing pH values from fossil material where predictor species are often absent. Of the four methods examined here it is suggested that multiple regression of diatom preference groups is probably the most suitable for pH reconstructions from sediment cores.     

Inferring pH from diatoms: a comparison of old and new calibration methods

Two new methods for inferring pH from diatoms are presented. Both are based on the observation that the relationships between diatom taxa and pH are often unimodal. The first method is maximum likelihood calibration based on Gaussian logit response curves of taxa against pH. The second is weighted averaging. In a lake with a particular pH, taxa with an optimum close to the lake pH will be most abundant, so an intuitively reasonable estimate of the lake pH is to take a weighted average of the pH optima of the species present.Optima and tolerances of diatom taxa were estimated from contemporary pH and proportional diatom counts in littoral zone samples from 97 pristine soft water lakes and pools in Western Europe. The optima showed a strong relation with Hustedt's pH preference groups. The two new methods were then compared with existing calibration methods on the basis of differences between inferred and observed pH in a test set of 62 additional samples taken between 1918 and 1983. The methods were ranked in order of performance as follows (between brackets the standard error of inferred pH in pH units); maximum likelihood (0.63) > weighted averaging (0.71) = multiple regression using pH groups (0.71) = the Gasse & Tekaia method (0.71) > Renberg & Hellberg's Index B (0.83) » multiple regression using taxa (2.2). The standard errors are larger than those usually obtained from surface sediment samples. The relatively large standard may be due to seasonal variation and to the effects of other factors such as humus content. The maximum likelihood method is statistically rigorous and can in principle be extended to allow for additional environmental factors. It is computer intensive however. The weighted averaging approach is a good approximation to the maximum likelihood method and is recommended as a practical and robust alternative.     

Recent ecological and biogeochemical changes in alpine lakes of Rocky Mountain National Park (Colorado, USA): a response to anthropogenic nitrogen deposition

Dated sediment cores from five alpine lakes (>3200 m asl) in Rocky Mountain National Park (Colorado Front Range, USA) record near‐synchronous stratigraphic changes that are believed to reflect ecological and biogeochemical responses to enhanced nitrogen deposition from anthropogenic sources. Changes in sediment proxies include progressive increases in the frequencies of mesotrophic planktonic diatom taxa and diatom concentrations, coupled with depletions of sediment δ¹⁵N and C : N values. These trends are especially pronounced since approximately 1950. The most conspicuous diatoms to expand in recent decades are Asterionella formosa and Fragilaria crotonensis. Down‐core species changes are corroborated by a year‐long sediment trap experiment from one of the lakes, which reveals high frequencies of these two taxa during autumn and winter months, the interval of peak annual limnetic []. Although all lakes record recent changes, the amplitude of stratigraphic shifts is greater in lakes east of the Continental Divide relative to those on the western slope, implying that most nitrogen enrichment originates from urban, industrial and agricultural sources east of the Rocky Mountains. Deviations from natural trajectories of lake ontogeny are illustrated by canonical correspondence analysis, which constrains the diatom record as a response to changes in nitrogen biogeochemistry. These results indicate that modest rates of anthropogenic nitrogen deposition are fully capable of inducing directional biological and biogeochemical shifts in relatively pristine ecosystems.     

Diatom species–environment relationships and inference models from Isachsen, Ellef Ringnes Island, Canadian High Arctic

Surface sediment diatom assemblages were examined from 26 freshwater sites near Isachsen (78°47N, 103°32W), Ellef Ringnes Island, a region of diverse and atypical water chemistry for high arctic sites. One hundred and sixty eight diatom taxa were identified from these samples, over 50% of which had not previously been recorded in the Canadian High Arctic. Variations in diatom assemblages were related to changes in measured environmental variables using multivariate techniques. Canonical correspondence analysis (CCA) indicated that five variables contributed significantly to explaining patterns of diatom variation (i.e., COND, DIC, Mn, TPF, TPU). The first CCA axis (=0.44) was primarily controlled by conductivity-related variables, while CCA axis 2 (=0.21) was related to particulate concentrations. Diatom-based inference models were generated for the reconstruction of conductivity (RMSEP_jack=0.32, r²_jack=0.76) and pH (RMSEP_jack=0.40, r²_jack=0.69). The strengths of these models indicate that it will be possible to reliably infer past trends in conductivity and pH from diatom assemblages preserved in dated sediment cores from the Isachsen region.     

Proton/electron stoichiometry of mitochondrialbc 1 complex. Influence of pH and transmembrane δpH

The effect of pH and transmembrane pH on the efficiency of the proton pump of the mitochondrialbc ₁ complex bothin situ and in the reconstituted state was studied. In both cases the H⁺/e ^– ratio for vectorial proton translocation by thebc ₁ complex respiring at the steady state, under conditions in which the transmembrane pH difference (pH) represents the only component of the proton motive force (p), was significantly lower than that measured under level flow conditions. The latter amounts, at neutral pH, to 1 (2 including the scalar H⁺ release). In the reconstituted system steady-state pH was modulated by changing the intravesicular buffer as well as the intra/extra-liposomal pH. Under these conditions the H⁺/e^– ratio varied inversely with the pH. The data presented show that pH exerts a critical control on the proton pump of thebc ₁ complex. Increasing the external pH above neutrality caused a decrease of the level flowH ⁺/e ^– ratio. This effect is explained in terms of proton/electron linkage inb cytochromes.     

pH effects on light dependence of the stoichiometry of proton influx and efflux to electron transport in spinach chloroplasts

Initial and steady state rates of proton transport at low light intensity have been measured and compared with steady state rates of electron transport in the pH range of 6.0–7.6 in envelope-free spinach chloroplasts. At pH 6–7, the H⁺/e^- values computed using the initial rate of proton transport varied with light intensity, from a value of 2 at low light to almost 5 at high light. In contrast, the H⁺/e^- values computed using the steady state rate of proton transport did not show a dependence on light intensity, having a constant value of 1.7±0.2. Likewise, at pH 7.6, the H⁺/e^- ratio, computed using either the initial or steady state rates of proton transport did not vary with light intensity but was constant at H⁺/e^-=1.7±0.1. Analysis of the light dependence of electron and proton transport allowed determination of (a) the quantam requirements of transport, (b) the rates of transport at light saturation, and (c) H⁺/e^- ratios for initial and steady state proton transport. Extrapolating the initial proton transport to zero light, we found that both H⁺/photon and H⁺/e^- values were not strongly dependent on pH, approaching a near constant value of 2.0. Using the initial rate of proton transport extrapolated to saturating light intensity we found the H⁺/e^- ratio to be strongly pH-dependent. We suggest that internal pH controls electron transport at high light intensities. The true stoichiometry is reflected only in measurements taken at low light using the initial proton transport data. Our findings and interpretation reconcile some conflicting data in the literature regarding the pH-dependence of the H⁺/e^- ratio and support the concept that internal pH controls noncyclic electron transport.Abbreviations Bicine N, N-bis [2-hydroxyethyl] glycine - HEPES N-2-hydroxy-ethylpiperazine-N-2-ethansulfonic acid - MES 2-(N-morpholino) ethanesulfonic acid     

Role of vacuolar adenosine triphosphatase in the regulation of cytosolic pH in hepatocytes

The responses of the cytosolic pH of hepatocytes in suspension to agents affecting the activity of vacuolar adenosine triphosphatase (V-ATPase) and Na/H exchange have been studied. Changes of cytosolic pH were determined both with dual-wavelength excitation (500/440 nm) of the fluorescence of 2,7-bis-(2-carboxyethyl)-5(and 6)-carboxyfluorescein and from the distribution of ¹⁴C-dimethyloxazolidinedione; both methods gave very similar results. Changes of vesicular pH were determined by comparing the fluorescence of fluorescein isothiocyanate-dextran and rhodamine B isothiocyanate-dextran taken up by endocytosis. Nitrate, which inhibits V-ATPase in isolated organelles, induced a concentration-dependent acidification of the cytosol and alkalinization of vesicles, with maximal effects at 25–37.5 mm in each case, indicating that V-ATPase contributes to removal of cytosolic protons. On continued exposure to nitrate, the acidification underwent an amiloride-inhibitable reversal. At the higher concentrations of NO ₃ ^– , both cytosolic acidification and vesicular alkalinization were reduced or absent. Bafilomycin A₁ caused alkalinization of vesicular pH; cytosolic acidification was not observed, possibly because of other ionic exchanges. Recovery of cytosolic pH from an acid load (2 min exposure to 5% CO₂) was sensitive to both 25 mm NO ₃ ^– and to ouabain. The pH dependence of the nitrate effect was tested with media of different pH; the activity was negligible at cytosolic pH 6.2 and rose to a maximum at cytosolic pH 7.3. Treatment of hepatocytes with 0.5–1.0 mm ouabain resulted in an initial alkalinization (0.5–2 min duration) of the cytosol, followed by a spontaneous reversal and, on occasion, further acidification. The alkalinization was blocked by 25 mm NO ₃ ^– , but not by 25 mm gluconate. The results suggest that the cytosolic alkalinization is caused by a stimulation of H⁺ uptake by V-ATPase activity. We conclude that V-ATPases make an important contribution to the regulation of the cytosolic pH of hepatocytes.This work was supported in part by National Institutes of Health B.R.S. Grant 507 RR05417 to Temple University.     

Impact of acidification on diatoms and chemistry of Dutch moorland pools

Old (ca 1920) and recent (1978) diatom assemblages from sixteen pristine moorland pools are compared by analysis of pH-spectra, diversity (Hill's index), (dis)similarity (number of species in common, Dyer dissimilarity) and principal component analysis. The pH-spectra of clear water pools indicate that the formerly wide range of pH (4–6) is very narrow now (3.7–4.6). No significant change of pH (ca 4.5) is observed in brown water moorland pools. Diversity significantly declines in clear water pools and has a tendency to rise in brown water pools. The number of species in common does not change. The Dyer dissimilarity significantly decreases in clear water lakes, no change is found in brown water lakes. The first principal component (PC 1) explains 61% of total variance between samples and is nearly completely determined byEunotia exigua. PC 1 has a strong positive correlation with both the absolute and relative sulfate concentration and other factors related to acidification (Ca²⁺, Al³⁺, Mg²⁺, electrical conductivity). PC1 has a negative correlation with factors characterizing humic acid waters (K MnO₄-consumption, Fe, (Na⁺ + K⁺)/ (Ca²⁺+Mg²⁺) etc.). Old samples have low scores on PC 1. Recent samples from clear waters have high scores on PC 1. The original variation, caused by regional factors, is replaced by a sulfate controled variation. The factors which are responsible for the recent differences in sulfate concentrations between pools are discussed (sulfate reduction, accumulation by dry deposition in adjacent pine forests and drought). Some observations contribute to the opinion that acidification may be considered as eutrophication and not as oligotrophication.