Toxicity of ammonia to algae in sewage oxidation ponds.

Ammonia, at concentrations over 2.0 mM and at pH values over 8.0, inhibits photosynthesis and growth of Scenedesmus obliquus, a dominant species in high-rate sewage oxidation ponds. Photosynthesis of Chlorella pyrenoidosa, Anacystis nidulans, and Plectonema boryanum is also susceptible to ammonia inhibition. Dark respiration and cell morphology were unaffected by any combination of pH and ammonia concentrations tested, thus limiting the apparent effect to inhibition of the normal function of the chloroplasts. Methylamine had the same effect as ammonia, and its penetration into the cells was found to be pH dependent. Therefore, the dependence of toxicity of amines to algae on pH apparently results from the inability to penetrate the cell membrane in the ionized form. When operated at 120-h detention time of raw wastewater, the high-rate oxidation pond maintained a steady state with respect to algal growth and oxygen concentration, and the concentration of ammonia did not exceed 1.0 mM. Shifting the pond to 48-h detention time caused an increase in ammonia concentration in the pond water to 2.5 mM, and the pond gradually turned anaerobic. Photosynthesis, which usually elevates the pH of the pond water to 9.0 to 10.0, could not proceed beyond pH 7.9 because of the high concentration of ammonia, and the algal population was washed out and reduced to a concentration that could maintain a doubling time of 48 h without photosynthesis bringing the pH to inhibitory levels. Under these conditions, the pH of the bond becomes a factor that limits the operational efficiency of the oxidation pond.     

Carbon limitation of biomass production in high-rate oxidation ponds

Theoretical considerations confirmed by outdoor experiments indicated carbon limitation of biomass production in high-rate oxidation ponds at certain seasonal and operational conditions. Apparently, free carbon dioxide concentration in the pond is the major determinant of carbonlimiting algal photosynthesis. High concentrations of free CO(2) are provided through bacterial respiration which is the main contributor to algal photosynthesis. At high photosynthetic activities and low organic loadings, free CO(2) concentrations are low; its flux into algal cells determines photosynthesis and biomass production rate in the pond.     

Effects of low concentrations of bisulfite-sulfite and nitrite on microorganisms.

A wide range of microorganisms was tested to determine their sensitivity to low concentrations of bisulfite-sulfite and nitrite, solubility products of SO2 and NO2, respectively. Photosynthesis by blue-green algae (cyanobacteria) was more strongly inhibited by 0.1 mM bisulfite-sulfite and 1 mM nitrite at pH 6.0 than photosynthesis by eucaryotic algae and respiration of bacteria, fungi, and protozoa. At pH 7.7, blue-green algae were still more sensitive to bisulfite-sulfite and nitrite than eucaryotic algae, but the toxicity of bisulfite-sulfite and nitrite decreased as the pH increased. Photosynthesis by Anabaena flos-aquae at pH 6.0 was inhibited 25% by a bisulfite-sulfite concentration of 10 micrometer and 15% by a nitrite concentration of 50 micrometer. Photosynthesis by the blue-green alga, Lyngbya sp., was not exceptionally sensitive to chlorate and thiosulfate. Acetylene-reducing activity of Beijerinckia indica was completely inhibited by 0.1 mM bisulfite-sulfite at pH 4.0, the suppression being decreased with increasing pH.     

Factors Affecting Infection of Scenedesmus obliquus by a Chytridium sp. in Sewage Oxidation Ponds

In a high-rate oxidation pond, 0.1 to 1.0% of the algal population of Scenedesmus obliquus was found to be infected by a chytrid. When suitable conditions developed, these infections burst into massive epidemics that killed most of the algae. The major factors triggering massive infections were optimal oxygen concentration and low concentrations of potassium and magnesium cations. The fungicide Benomyl was effective in preventing infection at a concentration of 1 mg/liter.     

Role of heterotrophic nutrition in growth of the alga Scenedesmus obliquus in high-rate oxidation ponds.

The green alga Scenedesmus obliquus readily adapted to heterotrophic growth in the dark, utilizing glucose as the sole carbon source. Heterotrophic cells differed significantly from photoautotrophic cells with respect to several physiological properties such as the rate of photoassimilation of CO2, rate of incorporation of glucose, and chlorophyll a concentration. Oxidation pond cells shared features common to both photoautotrophic and heterotrophic cells. Approximately 15 percent of oxidation pond algal carbon was derived from glucose assimilated directly without first being oxidized by bacteria. Bacteria seem to play a minor role in biological oxygen demand reduction in high-rate oxidation ponds, and their role is probably confined to degradation of biopolymers, thus producing substrates for algal consumption.     

Role of heterotrophic nutrition in growth of the alga Scenedesmus obliquus in high-rate oxidation ponds

The green alga Scenedesmus obliquus readily adapted to heterotrophic growth in the dark, utilizing glucose as the sole carbon source. Heterotrophic cells differed significantly from photoautotrophic cells with respect to several physiological properties such as the rate of photoassimilation of CO2, rate of incorporation of glucose, and chlorophyll a concentration. Oxidation pond cells shared features common to both photoautotrophic and heterotrophic cells. Approximately 15 percent of oxidation pond algal carbon was derived from glucose assimilated directly without first being oxidized by bacteria. Bacteria seem to play a minor role in biological oxygen demand reduction in high-rate oxidation ponds, and their role is probably confined to degradation of biopolymers, thus producing substrates for algal consumption.     

Influence of pH on Ammonia Accumulation and Toxicity in Halophilic, Methylotrophic Methanogens

We studied the effects of pH and ammonia concentration on the growth of three methanogens. These three halophilic, methylotrophic methanogens, Methanolobus bombayensis, Methanolobus taylorii, and Methanohalophilus zhilinaeae, grew at environmental pH ranges that overlapped with each other and spanned the pH range from 7.0 to 9.5. During growth they had reversed membrane pH gradients ((Delta)pH) at all pH values tested. The (Delta)pH was in the range of -0.4 to -0.9 pH units, with the cytosol being more acidic than the environmental pH. Methanohalophilus zhilinaeae had the most negative (Delta)pH (-0.9 pH units). These negative pH gradients resulted in the accumulation of ammonium (NH(inf4)(sup+)), and when grown at the highest external ammonia concentrations that allowed good growth, cells had cytosolic NH(inf4)(sup+) concentrations as high as 180 mM. The high concentrations of cytosolic NH(inf4)(sup+) were accompanied by greater (Delta)pH and lower concentrations of the major cytosolic cation K(sup+) (compared with cells grown in medium with only 5 mM ammonia). Methanolobus bombayensis and Methanolobus taylorii were more sensitive to total external ammonia at higher external pH values, but the inhibitory concentration of un-ionized ammonia that resulted in a 50% reduction of the growth rate was about 2 to 5 mM, regardless of the pH. This is consistent with growth inhibition by ammonia in other bacteria. However, Methanohalophilus zhilinaeae was more resistant to un-ionized ammonia than any other known organism. It had a 50% inhibitory concentration for un-ionized ammonia of 13 mM at pH 8.5 and 45 mM at pH 9.5. We examined the effects of pH on three ammonia-assimilating activities (glutamine synthetase, glutamate dehydrogenase, and alanine dehydrogenase) in cell lysates and found that the pH ranges were consistent with the observed ranges of intracellular pH.     

Simulating pH effects in an algal‐growth hydrodynamics model1

Models and numerical simulations are relatively inexpensive tools that can be used to enhance economic competitiveness through operation and system optimization to minimize energy and resource consumption, while maximizing algal oil yield. This work uses modified versions of the U.S. Environmental Protection Agency's Environmental Fluid Dynamics Code (EFDC) in conjunction with the U.S. Army Corp of Engineers' water‐quality code (CE‐QUAL) to simulate flow hydrodynamics coupled to algal growth kinetics. The model allows the flexibility of manipulating a host of variables associated with algal growth such as temperature, light intensity, and nutrient availability. pH of the medium is a newly added operational parameter governing algal growth that affects algal photosynthesis, differential availability of inorganic forms of carbon, enzyme activity in algae cell walls, and oil production rates. A single‐layer algal‐growth/hydrodynamic model without pH limitation was verified by comparing solution curves of algal biomass and phosphorus concentrations to an analytical solution. Media pH, now included in the model as a growth‐limiting factor, can be entered as a measured value or calculated based on CO₂ concentrations. Upon adding the ability to limit growth due to pH, physically reasonable results have been obtained from the model both with and without pH limitation. When the model was used to simulate algal growth from a pond experiment in the greenhouse, a least‐squares fitting technique yielded a maximum algal production (subsequently modulated by limitation factors) of 1.05 d^?1. Overall, the measured and simulated biomass concentrations in the greenhouse pond were in close agreement.     

Limitation of oxygenic photosynthesis and oxygen consumption by phosphate and organic nitrogen in a hypersaline microbial mat: a microsensor study

Microbial mats are characterized by high primary production but low growth rates, pointing to a limitation of growth by the lack of nutrients or substrates. We identified compounds that instantaneously stimulated photosynthesis rates and oxygen consumption rates in a hypersaline microbial mat by following the short-term response (c. 6 h) of these processes to addition of nutrients, organic and inorganic carbon compounds, using microsensors. Net photosynthesis rates were not stimulated by compound additions. However, both gross photosynthesis and oxygen consumption were substantially stimulated (by a minimum of 25%) by alanine (1 mM) and glutamate (3.5 mM) as well as by phosphate (0.1 mM). A low concentration of ammonium (0.1 mM) did not affect photosynthesis and oxygen consumption, whereas a higher concentration (3.5 mM) decreased both process rates. High concentrations of glycolate (5 mM) and phosphate (1 mM) inhibited gross photosynthesis but not oxygen consumption, leading to a decrease of net photosynthesis. Photosynthesis was not stimulated by addition of inorganic carbon, nor was oxygen consumption stimulated by organic compounds like glycolate (5 mM) or glucose (5 mM), indicating that carbon was efficiently cycled within the mat. Photosynthesis and oxygen consumption were apparently tightly coupled, because stimulations always affected both processes to the same extent, which resulted in unchanged net photosynthesis rates. These findings illustrate that microsensor techniques, due to their ability to quantify all three processes, can clarify community responses to nutrient enrichment studies much better than techniques that solely monitor net fluxes.     

Ammonium Limitation Results in the Loss of Ammonia-Oxidizing Activity in Nitrosomonas europaea

The effects of limiting concentrations of ammonium on the metabolic activity of Nitrosomonas europaea, an obligate ammonia-oxidizing soil bacterium, were investigated. Cells were harvested during late logarithmic growth and were incubated for 24 h in growth medium containing 0, 15, or 50 mM ammonium. The changes in nitrite production and the rates of ammonia- and hydroxylamine-dependent oxygen consumption were monitored. In incubations without ammonium, there was little change in the ammonia oxidation activity after 24 h. With 15 mM ammonium, an amount that was completely consumed, there was an 85% loss of the ammonia oxidation activity after 24 h. In contrast, there was only a 35% loss of the ammonia oxidation activity after 24 h in the presence of 50 mM ammonium, an amount that was not consumed to completion. There was little effect on the hydroxylamine oxidation activity in any of the incubations. The loss of ammonia oxidation activity was not due to differences in steady-state levels of ammonia monooxygenase (AMO) mRNA (amoA) or to degradation of the active site-containing subunit of AMO protein. The incubations were also conducted at a range of pH values to determine whether the loss of ammonia oxidation activity was correlated to the residual ammonium concentration. The loss of ammonia oxidation activity after 24 h was less at lower pH values (where the unoxidized ammonium concentration was higher). When added in conjunction with limiting ammonium, short-chain alkanes, which are alternative substrates for AMO, prevented the loss of ammonia oxidation activity at levels corresponding to their binding affinity for AMO. These results suggest that substrates of AMO can preserve the ammonia-oxidizing activity of N. europaea in batch incubations by protecting either AMO itself or other molecules associated with ammonia oxidation.     

Circadian rhythm and fast responses to blue light of photosynthesis in Ectocarpus (Phaeophyta,Ectocarpales)

Photosynthesis of Ectocarpus siliculosus (Dillwyn) Lyngb. under continuous saturating red irradiation follows a circadian rhythm. Blue-light pulses rapidly stimulate photosynthesis with high effectiveness in the troughs of this rhythm but the effectiveness of such pulses is much lower at its peaks. In an attempt to understand how blue light and the rhythm affected photosynthesis, the effects of inorganic carbon on photosynthetic light saturation curves were studied under different irradiation conditions. The circadian rhythm of photosynthesis was apparent only at irradiances which were not limiting for photosynthesis. The same was found for blue-light-stimulated photosynthesis, although stimulation was observed also under very low red-light irradiances after a period of adaptation, provided that the inorganic-carbon concentration was not in excess. Double-reciprocal plots of light-saturated photosynthetic rates versus the concentration of total inorganic carbon (up to 10 mM total inorganic carbon) were linear and had a common constant for half-saturation (3.6 mM at pH 8) at both the troughs and the peaks of the rhythm and before and after blue-light pulses. Only at very low carbon concentrations was a clear deviation found from these lines for photosynthesis at the rhythm maxima (red and blue light), which indicated that the strong carbon limitation specifically affected photosynthesis at the peak phases of the rhythm. Very high inorganic carbon concentrations (20 mM) in the medium diminished the responses to blue light, although they did not fully abolish them. The kinetics of the stimulation indicate that the rate of photosynthesis is affected by two blue-light-dependent components with different time courses of induction and decay. The faster component seemed to be at least partially suppressed at red-light irradiances which were not saturating for photosynthesis. Lowering the pH of the medium had the same effects as an increase of the carbon concentration to levels of approx. 10 mM. This indicates that Ectocarpus takes up free CO₂ only and not bicarbonate, although additional physiological mechanisms may enhance the availability of CO₂.Abbreviation TIC total inorganic carbon     

Phototrophs in high-iron-concentration microbial mats: physiological ecology of phototrophs in an iron-depositing hot spring

At Chocolate Pots Hot Springs in Yellowstone National Park the source waters have a pH near neutral, contain high concentrations of reduced iron, and lack sulfide. An iron formation that is associated with cyanobacterial mats is actively deposited. The uptake of [(14)C]bicarbonate was used to assess the impact of ferrous iron on photosynthesis in this environment. Photoautotrophy in some of the mats was stimulated by ferrous iron (1.0 mM). Microelectrodes were used to determine the impact of photosynthetic activity on the oxygen content and the pH in the mat and sediment microenvironments. Photosynthesis increased the oxygen concentration to 200% of air saturation levels in the top millimeter of the mats. The oxygen concentration decreased with depth and in the dark. Light-dependent increases in pH were observed. The penetration of light in the mats and in the sediments was determined. Visible radiation was rapidly attenuated in the top 2 mm of the iron-rich mats. Near-infrared radiation penetrated deeper. Iron was totally oxidized in the top few millimeters, but reduced iron was detected at greater depths. By increasing the pH and the oxygen concentration in the surface sediments, the cyanobacteria could potentially increase the rate of iron oxidation in situ. This high-iron-content hot spring provides a suitable model for studying the interactions of microbial photosynthesis and iron deposition and the role of photosynthesis in microbial iron cycling. This model may help clarify the potential role of photosynthesis in the deposition of Precambrian banded iron formations.     

Control of ammonia distribution ratio across the liver cell membrane and of ureogenesis by extracellular pH

The mechanisms involved in ammonia uptake by rat liver cells and the effects of changes in extracellular pH have been investigated in vivo and in vitro. When NH4Cl solutions were infused in the hepatic portal vein, ammonia uptake by the liver was practically quantitative up to about 1 mM in afferent blood. Ammonia transfer into hepatocytes was extremely rapid: for 2 mM ammonia in external medium, the intracellular concentration reached 5 mM within 10 s. Comparatively, [14C]methylamine influx was slower and the cell concentrations did not reach a steady-state level, probably in relation with diffusion into the acidic lysosomal compartment. Intracellular accumulation of ammonia was dependent on the delta pH across the plasma membrane: the distribution ratio (internal/external) was about 1 for an external pH of 6.8 and about 5 at pH 8. Urea synthesis was maximal at physiological pH and markedly declined at pH 7.05. This inhibition was not affected by manipulation of bicarbonate concentrations in the medium, down to 10 mM. Additional inhibition of ureogenesis by 100 microM acetazolamide was also observed, particularly at low concentrations of bicarbonate in the medium. Inhibition of ureogenesis when extracellular pH is decreased could be ascribed to a lower availability of the NH3 form. Assuming that NH3 readily equilibrates between the various compartments, the availability of free ammonia for carbamoyl-phosphate synthesis could be tightly dependent on extracellular pH.     

低强度超声波抑制铜绿微囊藻生长的研究

铜绿微囊藻(Microcystis aeruginosa)是常见的水华蓝藻,会对湖库的生态环境造成严重危害.室内研究了低强度超声波在不同藻生长时相、藻细胞浓度、水体pH、水温和二次超声条件下对铜绿微囊藻的抑制效果.结果表明:藻细胞浓度和pH对超声抑藻效果无明显影响,藻生长时相、水温和超声次数对超声抑藻效果有明显影响.15℃、20℃和25℃时的超声抑藻效果好于30℃和35℃,不经超声作用40℃高温即已不利于藻细胞生长;低强度超声对延滞期、稳定期和衰退期铜绿微囊藻抑制效果好于指数生长期铜绿微囊藻;二次超声可有效延长残余藻细胞的恢复时间,稳定抑藻效果.     

Phototrophs in High-Iron-Concentration Microbial Mats: Physiological Ecology of Phototrophs in an Iron-Depositing Hot Spring

At Chocolate Pots Hot Springs in Yellowstone National Park the source waters have a pH near neutral, contain high concentrations of reduced iron, and lack sulfide. An iron formation that is associated with cyanobacterial mats is actively deposited. The uptake of [¹⁴C]bicarbonate was used to assess the impact of ferrous iron on photosynthesis in this environment. Photoautotrophy in some of the mats was stimulated by ferrous iron (1.0 mM). Microelectrodes were used to determine the impact of photosynthetic activity on the oxygen content and the pH in the mat and sediment microenvironments. Photosynthesis increased the oxygen concentration to 200% of air saturation levels in the top millimeter of the mats. The oxygen concentration decreased with depth and in the dark. Light-dependent increases in pH were observed. The penetration of light in the mats and in the sediments was determined. Visible radiation was rapidly attenuated in the top 2 mm of the iron-rich mats. Near-infrared radiation penetrated deeper. Iron was totally oxidized in the top few millimeters, but reduced iron was detected at greater depths. By increasing the pH and the oxygen concentration in the surface sediments, the cyanobacteria could potentially increase the rate of iron oxidation in situ. This high-iron-content hot spring provides a suitable model for studying the interactions of microbial photosynthesis and iron deposition and the role of photosynthesis in microbial iron cycling. This model may help clarify the potential role of photosynthesis in the deposition of Precambrian banded iron formations.     

Acid Water Interferes with Salamander-Green Algae Symbiosis during Early Embryonic Development

Abstract The inner egg capsule of embryos of the yellow-spotted salamander (Ambystoma maculatum) are routinely colonized by green algae, such as Oophila amblystomatis, that supply O(2) in the presence of light and may consume nitrogenous wastes, forming what has been proposed to be a mutualistic relationship. Given that A. maculatum have been reported to breed in acidic (pH <5.0) and neutral lakes, we hypothesized that low water pH would negatively affect these symbiotic organisms and alter the gradients within the jelly mass. Oxygen gradients were detected within jelly masses measured directly in a natural breeding pond (pH 4.5-4.8) at midday in full sunlight. In the lab, embryo jelly masses reared continuously at pH 4.5 had lower [Formula: see text] and higher ammonia levels relative to jelly masses held at pH 8.0 (control). Ammonia and lactate concentrations in embryonic tissues were approximately 37%-93% higher, respectively, in embryos reared at water pH 4.5 compared with pH 8.0. Mass was also reduced in embryos reared at pH 4.5 versus pH 8.0. In addition, light conditions (24 h light, 12L∶12D, or 24 h dark) and embryonic position (periphery vs. center) in the jelly mass affected [Formula: see text] but not ammonia gradients, suggesting that algal symbionts generate O(2) but do not significantly impact local ammonia concentrations, regardless of the pH of the water. We conclude that chronic exposure to acidic breeding ponds had a profound effect on the microenvironment of developing A. maculatum embryos, which in turn resulted in an elevation of potentially harmful metabolic end products and inhibited growth. Under acidic conditions, the expected benefit provided by the algae to the salamander embryo (i.e., high O(2) and low ammonia microenvironment) is compromised, suggesting that the A. maculatum-algal mutualism is beneficial to salamanders only at higher water pH values.     

The effect of pH on the toxicity of ammonia to a murine hybridoma.

The growth inhibition of a murine hybridoma mediated by ammonium chloride was shown to vary with the pH of the culture medium. Values for the initial media concentration causing 50% growth inhibition (IC50) ranged from 4 mM to 7.6 mM as the pH was reduced from 7.8 to 6.8. A significant negative correlation was observed between the IC50 and the NH3 concentration of the medium, suggesting that ammonia and not ammonium may be the toxic species in the culture medium. The optimum initial pH for cell growth was 7.4. However, this optimum shifts to lower pH as ammonia accumulates in culture as a metabolic by-product. This suggests that in order to obtain high cell yields, it may be beneficial to adopt a culture strategy of lowering pH during cell growth to offset the inhibitory effects of accumulated ammonia.     

Effect of Hard Detergents on Algae in a High-Rate-Oxidation Pond

Regular concentrations of hard detergents in domestic wastewater do not affect algal growth in a high-rate-oxidation pond. The addition of nonionic hard detergent at concentrations above 60 mg/liter decreased the algal concentration in the batch culture, and complete lysis of algal cells was observed within a few days at a detergent concentration of 100 mg/liter.     

Modification of the interaction between<Emphasis Type="Italic">Escherichia coli</Emphasis> and bacteriophage in saline sediment

The effects of various combinations of light intensity, oxygen concentration, and CO₂ concentration on photosynthesis and growth in several algal types were studied. The results suggest the following. (1) Different algae show different responses to high oxygen concentrations and high light intensities. (2) Inhibition of photosynthesis (CO₂ fixation and growth), if seen, increases with increasing oxygen concentration and with increasing light intensity (at light intensities greater than saturation). (3) The inhibition of net photosynthesis observed cannot be attributed to high light intensity alone. (4) The inhibition cannot be attributed to increased rates of excretion of organic materials under conditions of high oxygen concentration and high light intensity. (5) Increased concentrations of CO₂ can decrease the effect of high oxygen and light in some algae. (6) The decrease in net photosynthesis observed is probably the result of photorespiration. (7) The effect of light intensity, oxygen concentration, or CO₂ concentration on algal photosynthesis should not be studied without considering the effect of the other factors. Some implications of these results, as related to primary productivity measurements, are also discussed.     

Ammonia inhibition of the maximum growth rate (μm) of hydrogenotrophic methanogens at various pH-levels and temperatures

Summary A method for the quantitative measurement of the maximum growth rate (_m) of hydrogen-consuming methanogenic populations was applied to assess the toxicity of ammonia¹ under various pH and temperature conditions. The maximum uninhibited growth rate of the hydrogenotrophic population present in sludge from an industrial anaerobic wastewater treatment system appeared to be 0.126 h^-1 at pH=7 and 37°C. At 350 mM ammonia the maximum growth rate had decreased to almost half that value. At a temperature of 29°C the maximum growth rates in the ammonia range tested appeared to be approximately 60% of that at 37°C, while increasing ammonia concentrations caused a similar maximum growth rate decline. At 37°C an increase of the pH to 7.8 appeared to enhance ammonia inhibition of the maximum growth rate. Increased propionate concentrations (tested up to 60 mM) appeared to have no effect on ammonia inhibition.