The role of zooplankton in the intrabiocoenotic phosphorus cycle and factors affecting phosphorus excretion in a lake

The temporal and spatial excretion rates of specific size groups of limnetic zooplankton were studied by measuring changes in soluble reactive phosphorus following incubation. Animals were collected in the epilimnion and hypolimnion of Stonehouse Pond, New Hampshire from August, 1972 to July, 1973 and separated into > 0.308 and < 0.308 (mm) size groups. Temporal excretion rates varied considerably within groups, however, similar patterns were observed in both strata. Peak excretion rates were observed in the spring and fall with a low in winter months. In addition, smaller animals excreted at higher rates and excretion rates of both size groups in the epilimnion exceeded those in the hypolimnion.The phosphorus uptake of natural seston relative to the amount released by zooplankton is considered using mass balance equations and kinetic analyses. During the spring of 1973 excretion rates increased, however, this increase was offset by an even greater rise in the rate of phosphorus uptake by the seston. This suggests that during this period the phosphorus excretions of zooplankton were not sufficient to meet the amount of phosphorus being removed by the seston.The experimental work was based on a thesis submitted to the Graduate School of the University of New Hampshire in partial fulfillment of the requirements for the degree of Doctor of Philosophy.     

Orthophosphate turnover in East African lakes

Summary Turnover rates of ³²P–PO₄ and concentrations of orthophosphate as soluble reactive phosphorus (SRP) were measured in five East African waters. Rapid incorporation of ³²P–PO₄ by the seston and orthophosphate concentrations below the limit of detectibility were found in Lakes Elmenteita, Naivasha, and Naivasha Crater Lake. Turnover was slow and orthophosphate concentration high in both Lake Nakuru and the Crescent Island Crater basin of Lake Naivasha. Further experiments in Lake Nakuru indicated that colloidal binding of orthophosphate was limited and that particles retained by an 8.0 filter incorporated 66% as much tracer as particles retained by a 0.1 filter. These experiments strengthen our conclusion that a large quantity of orthophosphate is available for algal use in Lake Nakuru.     

Inorganic polyphosphate as an integral part of alkaline phosphatase preparations.

Alkaline phosphatase (from chicken intestinal sources) was shown to contain a considerable amount of polyanionic phosphorus which was released by basic digestion. The polyanionic phosphorus of alkaline phosphatase is not associated with protein or polyalcohols and does not exhibit a visible or ultraviolet absorption spectrum. Alkaline phosphatase and abiogenic inorganic polyphosphate were found to incorporate 32P-orthophosphate under similar experimental conditions. It has been previously reported that this enzyme will incorporate 32P-orthophosphate into its protein phosphoserine without the apparent concomitant utilization of an energy source. This reported phosphorylation was immediately reversible upon dilution of the phosphorylated enzyme with unlabelled orthophosphate, which indicates that the initial phosphorylation was an exchange reaction. These observations suggest that this polyanionic phosphorus from alkaline phosphatase may be inorganic polyphosphate.     

Regeneration of inorganic phosphorus and nitrogen from decomposition of seston in a freshwater sediment

The mineralization of phosphorus and nitrogen from seston was studied in consolidated sediment from the shallow Lake Arreskov (July and November) and in suspensions without sediment (July). In the suspension experiment, phosphorus and nitrogen were mineralized in the same proportions as they occurred in the seston. During the 30 days suspension experiment, 47 and 43% of the particulate phosphorus and nitrogen, respectively, was mineralized with constant rates.Addition of seston to the sediment had an immediate enhancing effect on oxygen uptake, phosphate and ammonia release, whereas nitrate release decreased due to denitrification. The enhanced rates lasted for 2–5 weeks, while the decrease in nitrate release persisted throughout the experiment. The increase in oxygen uptake (equivalent to 21% of the seston carbon) was, however, only observed in the July experiment. The release of phosphorus and nitrogen from seston decomposing on the sediment surface differed from the suspension experiments. Thus, between 91 and 111% of the phosphorus in the seston was released during the experiments. Due to opposite directed effects on ammonium and nitrate release, the resulting net release of nitrogen was relatively low.A comparison of C/N/P ratios in seston, sediment and flux rates indicated that nitrogen was mineralized faster than phosphorus and carbon. Some of this nitrogen was lost through denitrification and therefore not measurable in the flux of inorganic nitrogen ions. This investigation also suggests that decomposition of newly settled organic matter in sediments have indirect effects on sediment-water exchanges (e.g. by changing of redox potentials and stimulation of denitrification) that modifies the release of mineralized phosphate and nitrogen from the sediment.     

Rapid physiological assays for nutrient demand by the plankton. II. Phosphorus

Four bioassays for phosphorus demand were compared. ³²P-turnoverrates provided an integrated measure of transport capacity ofthe assemblage and ambient orthophosphate levels, but rateswere often not simply exponential and could not be accuratelyquantified by a single rate constant. ATP responses to phosphorusenrichment were transient and they poorly separated naturalplankton populations with very different degrees of P-deficiency.Alkaline phosphatase activity corresponded well with other measuresof phosphorus demand although low activities did not necessarilyimply non-limiting phosphorus conditions. Selective luxury accumulationof N and P by the seston from an N + P enrichment provided themost informative guide to the relative demand for the two elements.No single approach could be considered conclusive on its own,but in combination the assays provided a reliable measure ofphosphorus demand and deficiency within natural assemblagesof plankton.     

Starch Phosphorylation in Potato Tubers Proceeds Concurrently with de Novo Biosynthesis of Starch

The in vivo phosphorylation of starch was studied in Solanum tuberosum cv Dianella and Posmo. Small starch granules contain 25% more ester-bound phosphate per glucose residue than large starch granules. The degree of phosphorylation was found to be almost constant during tuber development. Isolated tuber discs synthesize starch from externally supplied glucose at a significant rate. Tuber discs supplied with glucose and [32P]orthophosphate incorporate radiolabeled phosphorus into the starch. The level of 32P incorporation is proportional to the amount of starch synthesized. The incorporation of 32P from orthophosphate is correlated to de novo synthesis of starch, since the incorporation of 32P is diminished upon inhibition of starch synthesis by fluoride. Based on the amount of [14C]glucose phosphate isolated after hydrolysis of purified starch from tuber discs incubated in the presence of [U-14C]glucose, approximately 0.5% of the glucose residues of the de novo-synthesized starch are phosphorylated. This value is in general agreement with the observed levels of phosphorus in starch accumulated during tuber development. Thus, the enzyme system responsible for starch phosphorylation is fully active in the isolated tuber discs, and the starch phosphorylation proceeds as an integrated part of de novo starch synthesis.     

Effects of condensed phosphates on plant growth and phosphorus uptake

Summary Four condensed phosphates with large and small molecules, including ring and chain structures, were equivalent to orthophosphate in terms of phosphorus uptake and the dry matter yield of ryegrass grown in two soils in pots. This equivalence was maintained at each of 6 cuts during two seasons.Increasing N rate greatly increased uptake of applied phosphorus but there was no differential effect with different sources of phosphorus, nor was there any interaction between source and application rate.Phosphorus was not leached at any time, indicating that the phosphates were rapidly adsorbed by the soils.Fairly rapid hydrolysis of all the condensed phosphates occurred in a soil of neutral pH but a slower hydrolysis seemed to occur in an acid soil.There was a good correlation between the phosphorus uptake by ryegrass and the additional orthophosphate released by an acid hydrolysis of soil extracts.     

Impact of weather variability on spatial and seasonal dynamics of dissolved and suspended nutrients in water column of meromictic Lake Shira

Depths of thermocline and the redox zone, concentrations of dissolved and suspended carbon, and nitrogen and phosphorus in seston were measured in the pelagial of the saline meromictic Lake Shira (southern Siberia) in different years (2007–2011); the relationship of the values for those parameters with air-temperature variations was assessed. Positive correlations between both air temperatures in the previous year and the depth of the redox zone in winter and air temperature in April and the thermocline depth in summer were revealed. In the mixolimnion, the ratio of total nitrogen to total phosphorus almost always exceeded the Redfield ratio (16: 1); seston deficiency both in nitrogen and phosphorus was monitored in different seasons and at various depths. The amount of seston in the mixolimnion in summer almost doubled the amount of seston in winter and was directly related with the depth of the thermocline. In the monimolimnion, seston was rich in nitrogen and phosphorus. The amount of seston in the monimolimnion varied in different years and depended both on the air temperature in the previous year and the size of the zone.     

Phosphonate utilization by bacteria.

Bacteria able to use at least one of 13 ionic alkylphosphonates of O-alkyl or O,O-dialkyl alkylphosphonates as phosphorus sources were isolated from sewage and soil. Four of these isolates used 2-aminoethylphosphonic acid (AEP) as a sole carbon, nitrogen, and phosphorus source. None of the other phosphonates served as a carbon source for the organisms. One isolate, identified as Pseudomonas putida, grew with AEP as its sole carbon, nitrogen, and phosphorus source and released nearly all of the organic phosphorus as orthophosphate and 72% of the AEP nitrogen as ammonium. This is the first demonstration of utilization of a phosphonoalkyl moiety as a sole carbon source. Cell-free extracts of P. putida contained an inducible enzyme system that required pyruvate and pyridoxal phosphate to release orthophosphate from AEP; acetaldehyde was tentatively identified as a second product. Phosphite inhibited the enzyme system.     

Phosphorus availability and alkaline phosphatase activities in two Israeli fishponds

1. Alkaline phosphatase activities and the release of orthophosphate from endogenous substrates by these enzymes were measured in waters from two commercial fishponds in the watershed area of Lake Kinneret (Northern Israel). These data were compared with results from the lake at seasons of adequate or limited phosphorus supply. In the fishponds, high Relative Phosphatase Activity ratios (>2) and relatively large amounts of orthophosphate extracted from plankton by autoclaving (average 27% of readily available phosphorus) indicated adequate, or even excess, levels of phosphorus availability despite elevated pond productivity (2 to 3 tons carp/ha/yr). Therefore, we suggest that decreasing routine phosphorus fertilization of these ponds would not affect overall productivity but would eventually lower the amounts of phosphorus reaching Lake Kinneret.
2. In general, the R. P. A. ratio may be a useful index to evaluate phosphorus availability for a wide range of natural waters. Values for this ratio of <1 and >2 appear indicative of limited or adequate phosphorus availability respectively.
3. Three sources of orthophosphate, (Pi), readily available to phytoplankton, are indicated: (1) enzymatically released Pi, (2) Pi in intracellular pools and (3) Pi initially present in the water. Although the first source is always important, relatively greater amounts of Pi are contributed by the other fractions in situations of plentiful phosphorus availability.
4. Activity of free dissolved phosphatases was found in filtered samples of fishpond water. However, neither these enzymes or added phosphatases released significant amounts of Pi from the dissolved organic phosphorus compounds in the filtered water.

     

Enzymatic release of phosphate in water from subarctic lakes in Northern Sweden

Enzymatic phosphate release was studied in water from three small lakes in the subarctic region of northern Sweden. Free dissolved hydrolytic enzymes were shown to release considerable amounts of phosphate from autoclaved lake water. It was possible to show that phosphate was enzymatically released from particulate matter by exposure of autoclaved, ³²P-labelled plankton to lake enzymes followed by characterization of degradation products with gel chromatography. Seston turned out to be vitally important as a phosphorus source for planktic organisms since plankton inoculated in lake water freed from seston were shown to produce high activities of phosphatase.     

Limax Amoebae in Public Swimming Pools of Albany, Schenectady, and Rensselaer Counties, New York: Their Concentration, Correlations, and Significance

A survey was conducted on 30 halogenated public swimming pools, located in Albany, Schenectady, and Rensselaer counties, to determine their open-water limax amoeba densities. Six were outdoor pools. Other variables measured were the standard plate count, total seston, free residual chlorine or bromine, total alkalinity, total hardness, orthophosphate, total soluble phosphorus, specific conductance, pH, temperature, and several engineering parameters including the rate and type of filtration as well as a saturation index. Amoebae were isolated on agar plates at 37°C using heat-killed bacterial suspensions of Enterobacter cloacae or Escherichia coli. Most probable number estimates of amoebic densities ranged from not detectable (<0.01) to 110 amoebae per liter. The median concentration of amoebae was 0.9/liter. Eighty percent of the pools examined had less than 5 amoebae per liter. Significant correlations (P < 0.05) were found between amoebic densities and the log₁₀ of the standard plate count, orthophosphate, and total soluble phosphorus. No significant difference was found between amoebic densities in outdoor and indoor pools. Preliminary tests for the presence of the human pathogen Naegleria fowleri were inconclusive.     

Cell wall metabolism in Bacillus subtilis subsp. niger: effects of changes in phosphate supply to the culture

Chemostat cultures of Bacillus subtilis subsp. niger WM were exposed to changes in the availability of phosphorus by means of a resuspension technique. Responses in wall metabolism were recorded by measuring the amounts of peptidoglycan and anionic polymers (teichoic or teichuronic acid) in the wall and extracellular fluid fractions. With respect to the wall composition, the effect of a change in orthophosphate supply was a complete shift in the nature of the anionic polymer fraction, the polymer originally present in the walls ("old" polymer) being replaced by the alternative ("new") anionic polymer. The peptidoglycan content of the walls remained constant. It was concluded that the incorporation of old polymer was completely blocked from the moment the orthophosphate supply was changed. However, from a measurement of the total amount of polymer in the whole culture during the course of the experiments, it was evident that synthesis of old polymer continued, but it was secreted. Synthesis of the new polymer started immediately, and it was incorporated exclusively into the wall. During adaption of the cells to the new environment, wall turnover continued in an identical fashion to that extant in steady-state cultures. It was concluded that the primary adaptive response to a change in orthophosphate supply occurred through a mechanism interacting with polymer incorporation and thus at the level of wall assembly at the membrane.     

Evidence of phosphorus-limited individual and population growth of Daphnia in a Canadian Shield lake

We performed bag experiments in a Canadian Shield lake with generally high seston (suspended food particles mainly composed of algae) carbon (C):phosphorus (P) ratios, and investigated the responses of individual and population growth of herbivorous Daphnia dentifera on their abundance with (+P) and without (−P) a phosphorus enrichment to lake water. In both treatments, increased abundance of D. dentifera reduced seston C concentration and was accompanied by decreases in population and individual growth rates. However, P-enrichment increased seston P concentration and then reduced seston C:P ratio from 400–700 to ca 100 (by atoms). As a result, both individual and population growth rates were significantly higher in the +P treatment at all animal abundances even though seston C concentrations were similar between the treatments. The magnitude of the growth enhancement by the P-enrichment was independent of animal abundance. Stepwise regression analyses revealed that 71 and 90% of the variance in the population and individual growth rates, respectively, were explained by seston C and P concentrations, and that the contribution of the seston P concentration was roughly the same as that of seston C. Such joint effects of seston C and P indicate that food quality (P content) as well as food quantity (C concentration) can influence Daphnia not only at the level of individual growth but also at the level of population dynamics in P-limited lakes. Our results thus strongly corroborate the hypothesis that the population development of a key herbivore Daphnia in P-limited Canadian Shield lakes is inhibited by the direct effects of P-limited food on individual growth, which weaken the strength of trophic cascading interactions starting from piscivorous fish through planktivorous fish and zooplankton to algae.     

Nutritional quality of seston for the freshwater herbivore Daphnia galeata × hyalina: biochemical versus mineral limitations

The quality of natural seston as food for zooplankters can be highly variable. Thus far, experimental evidence on the factors affecting food quality under natural conditions is scarce. Hence, in this study, we set out to investigate how Daphnia galeata 2 hyalina responded to qualitative variation in natural seston. This was done in laboratory experiments where we supplement natural seston from a mesotrophic lake with dissolved phosphorus and emulsions of highly unsaturated "essential" fatty acids. The growth rate of juveniles increased upon the supply of both phosphorus and fatty acids. These results suggest that these phosphorus and highly unsaturated fatty acids are substitutable and thus challenge our existing interpretation/understanding of how herbivore growth is "limited".     

Phosphatases; origin,characteristics and function in lakes

Phosphatases catalyze the liberation of orthophosphate from organic phosphorus compounds. The total phosphatase activity in lake water results from a mixture of phosphatases localized on the cell surfaces of algae and bacteria and from dissolved enzymes supplied by autolysis or excretion from algae, bacteria and zooplankton. External lake water phosphatases usually have pH optima in the alkaline region. Acid phosphatases generally seem to be active in the internal cell metabolism. The synthesis of external alkaline phosphatases is often repressed at high phosphate concentrations and derepressed at low phosphate concentrations. Phosphatase activity has therefore been used as a phosphorus deficiency indicator in algae and in natural plankton populations. The possibilities for this interpretation of phosphatase activity in lake water are limited, however, and this is discussed. The in situ hydrolysis capacity, i.e. the rate by which orthophosphate is released from natural substrates, is unknown. However, we advocate that this process is important and that the rate of substrate supply, rather than phosphatase activity, limits the enzymatic phosphate regeneration.     

淡水中型浮游动物群体与悬浮物碳氮磷比耦联研究

通过对湖北省13个湖库悬浮物和中型浮游动物群体碳氮磷比的分析, 探讨了浮游动物与食物之间的元素耦联和不平衡性是如何随食物质量的改变而变化, 以及这种不平衡性对生态系统浮游动植物间能量传递效率的影响。结果显示: 中型浮游动物C﹕N比和C﹕P比分别随悬浮物C﹕N比和C﹕P比的增加而变大。悬浮物-中型浮游动物之间的元素不平衡性不仅与悬浮物C﹕N比显著正相关(r=0.97, P<0.001), 且与悬浮物C﹕P比也显著正相关(r=0.81, P=0.001)。浮游动植物之间的能量传递效率与悬浮物-中型浮游动物C﹕P比差值呈显著负相关(r= –0.58, P=0.037), 但与其C﹕N比差值关系不显著(P>0.05)。此外, 中型浮游动物生物量也随其与悬浮物之间C﹕P比差值的增大而下降(r= –0.59, P=0.033)。这些结果表明, 悬浮物碳氮磷比能显著影响中型浮游动物群体的碳氮磷比组成, 两者之间的元素不平衡性随前者的增加而变大并进而降低能量向后者的传递。     

Identification of 5'-Adenylylimidodiphosphate-Hydrolyzing Enzyme Activity in Rabbit Taste Bud Cells Using X-Ray Microanalysis

X-ray microanalysis has been used to characterize the enzyme activity hydrolyzing the ATP analogue 5'-adenylylimidodiphosphate (AMP-PNP) in taste bud cells. Rabbit foliate papillae fixed with paraformaldehyde and glutaraldehyde were incubated cytochemically with AMP-PNP as the substrate and lead ion as capture agent. The reaction product which appeared on the microvilli of taste bud cells was examined using an energy dispersive X-ray microanalyzer connected to an analytical electron microscope. The X-ray spectrum thus obtained was compared with that obtained from the product obtained from the demonstration of ATPase activity. Comparison of the phosphorus/lead ratios in the two products showed that twice as much phosphorus was released from an AMP-PNP molecule by the activity in question compared with that released from an ATP molecule by ATPase activity. This indicates that the enzyme hydrolyzes AMP-PNP into AMP and imidodiphosphate and that the enzyme is adenylate cyclase or ATP pyrophosphohydrolase, which possesses a similar hydrolytic property, but not ATPase or alkaline phosphatase, which hydrolyzes AMP-PNP into ADP-NHj and orthophosphate. This paper provides an example of the use of X-ray microanalysis as a tool for enzyme distinction. The method is applicable to a variety of enzymes and tissues.     

Identification of 5'-adenylylimidodiphosphate-hydrolyzing enzyme activity in rabbit taste bud cells using X-ray microanalysis

X-ray microanalysis has been used to characterize the enzyme activity hydrolyzing the ATP analogue 5'-adenylylimidodiphosphate (AMP-PNP) in taste bud cells. Rabbit foliate papillae fixed with paraformaldehyde and glutaraldehyde were incubated cytochemically with AMP-PNP as the substrate and lead ion as capture agent. The reaction product which appeared on the microvilli of taste bud cells was examined using an energy dispersive X-ray microanalyzer connected to an analytical electron microscope. The X-ray spectrum thus obtained was compared with that obtained from the product obtained from the demonstration of ATPase activity. Comparison of the phosphorus/lead ratios in the two products showed that twice as much phosphorus was released from an AMP-PNP molecule by the activity in question compared with that released from an ATP molecule by ATPase activity. This indicates that the enzyme hydrolyzes AMP-PNP into AMP and imidodiphosphate and that the enzyme is adenylate cyclase or ATP pyrophosphohydrolase, which possesses a similar hydrolytic property, but not ATPase or alkaline phosphatase, which hydrolyzes AMP-PNP into ADP-NH2 and orthophosphate. This paper provides an example of the use of X-ray microanalysis as a tool for enzyme distinction. The method is applicable to a variety of enzymes and tissues.     

Biological Uptake of Phosphorus by Activated Sludge

The ability of activated sludge to remove phosphates was studied by adding carrier-free (32)P to raw sewage and measuring incorporation of the radioactivity into the cells over a period of time. Radioisotope determinations indicated that 48% of the (32)P radioactivity was removed by 12 hr. However, chemical methods indicated that only 30% of the orthophosphate apparently disappeared from the sewage during this period. Experiments with sludge prelabeled with (32)P indicated that considerable phosphate turnover occurred. The cells released large amounts of radioactivity as they were incorporating fresh phosphates. Starvation in isotonic saline for 18 hr caused the sludge to dump phosphate. When introduced into fresh sewage containing (32)P, the starved sludge removed about 60% of the radioactivity in 6 hr with little phosphate turnover. The ability of sludge to remove (32)P was inhibited approximately 83% by 10(-3)m 2,4-dinitrophenol. This inhibition was at the expense of the cell fraction that contained ribonucleic acid and deoxyribonucleic acid. The sludge cells released orthophosphate when exposed to the chemical agent. Experiments using (45)Ca indicated that calcium phosphate precipitation plays a minor role in phosphate removal under our experimental conditions.