Global transcriptional responses of the toxic cyanobacterium, Microcystis aeruginosa, to nitrogen stress, phosphorus stress, and growth on organic matter

Whole transcriptome shotgun sequencing (RNA-seq) was used to assess the transcriptomic response of the toxic cyanobacterium Microcystis aeruginosa during growth with low levels of dissolved inorganic nitrogen (low N), low levels of dissolved inorganic phosphorus (low P), and in the presence of high levels of high molecular weight dissolved organic matter (HMWDOM). Under low N, one third of the genome was differentially expressed, with significant increases in transcripts observed among genes within the nir operon, urea transport genes (urtBCDE), and amino acid transporters while significant decreases in transcripts were observed in genes related to photosynthesis. There was also a significant decrease in the transcription of the microcystin synthetase gene set under low N and a significant decrease in microcystin content per Microcystis cell demonstrating that N supply influences cellular toxicity. Under low P, 27% of the genome was differentially expressed. The Pho regulon was induced leading to large increases in transcript levels of the alkaline phosphatase phoX, the Pst transport system (pstABC), and the sphX gene, and transcripts of multiple sulfate transporter were also significantly more abundant. While the transcriptional response to growth on HMWDOM was smaller (5–22% of genes differentially expressed), transcripts of multiple genes specifically associated with the transport and degradation of organic compounds were significantly more abundant within HMWDOM treatments and thus may be recruited by Microcystis to utilize these substrates. Collectively, these findings provide a comprehensive understanding of the nutritional physiology of this toxic, bloom-forming cyanobacterium and the role of N in controlling microcystin synthesis.

Historical nitrogen and phosphorus loadings to the northern Gulf of Mexico

Primary productivity in many coastal systems is nitrogen (N) limited; although, phytoplankton productivity may be limited by phosphorus (P) seasonally or in portions of an estuary. Increases in loading of limiting nutrients to coastal ecosystems may lead to eutrophication (Nixon 1996). Anthropogenically enhanced eutrophication includes symptoms such as loss of seagrass beds, changes in algal community composition, increased algal (phytoplankton) blooms (Richardson et al. 2001), hypoxic or anoxic events, and fish kills (Bricker et al. 2003).

The influence of nitrogen and phosphorus on the growth of a diatom Skeletonema costatum (Greville) Cleve

Nitrogen and phosphorus requirements of a chain-forming diatom, Skeletonema costatum (Greville) Cleve, collected from Yatsushiro Sea, Japan, were investigated in a laboratory culture experiment. Sodium nitrate and sodium glycerophosphate were used as nitrogen and phosphorus sources, respectively. Cultures were grown in modified Provasoli's ASP2NTA medium (Provasoli et al. 1957) at 25±1°C, light intensity 60 µE mˉ² secˉ¹ and photoperiod 12:12-h, L:D cycle. Optimum growth was observed at nitrate concentrations of 3-10 mglˉ¹ and phosphate concentrations of 1.5-15 mglˉ¹. Adequate growth was also found at the nitrate concentration of up to as high as 300 mglˉ¹. Significantly poorer growth was found at lower nitrate (<3.0 mglˉ¹) and higher phosphate (>15 mglˉ¹) concentrations. From the present study, it is concluded that S. costatum can grow well at wide ranges of nitrate concentrations but is sensitive to higher phosphate concentrations.

Microwave power effects on the properties of phosphorus-doped SiC:H films prepared using ECR-CVD

There has been a growing interest in hydrogenated silicon carbide films (SiC:H) prepared using the electron cyclotron resonance-chemical vapour deposition (ECR-CVD) technique. Using the ECR-CVD technique, SiC:H films have been prepared from a mixture of methane, silane and hydrogen, with phosphine as the doping gas. The effects of changes in the microwave power (from 150 to 900 W) on the film properties were investigated in a series of phosphorus-doped SiC:H films. In particular, the changes in the deposition rate, optical bandgap, activation energy and conductivity were investigated in conjunction with results from Raman scattering and Fourier transform infra-red (FTIR) analysis. It was found that increase in the microwave power has the effect of enhancing the formation of the silicon microcrystalline phase in the amorphous matrix of the SiC:H films. This occurs in correspondence to a rapid increase in the conductivity and a reduction in the activation energy, both of which exhibit small variations in samples deposited at microwave powers exceeding 500 W. Analysis of IR absorption results suggests that hydrogen is bonded to silicon in the Si-H stretching mode and to carbon in the sp3 CHn rocking/wagging and bending mode in films deposited at higher microwave powers.

Effects of dietary phosphorus and zinc levels on growth artd bone mineralization in fingerlings of rainbow trout, Oncorhynchus mykiss

A laboratory based 2 x 2 factorial experiment was conducted to investigate the influences of dietary phosphorus and zinc levels on growth and bone mineralization in fingerlings of rainbow trout for 21 weeks. Two levels of phosphorus (19 and 30 mg/g) and two levels of zinc (55 and 103 Ag/g) in the dry diets were tested. Duplicate tanks of 30 rainbow trout (average weight 1.56 ± 0.24 g) per 60L glass tank were fed experimental diets three times a day to apparent satiation level at 15 to 24°C water temperature. The results of the present study demonstrated that dietary phosphorus supplementation influenced the growth and bone mineralization whereas zinc levels significantly (p<0.05) influenced bone mineralization in rainbow trout. Further investigations in this area with different size and age groups of this fish are broadly needed.

Effects of iso-nitrogenous and iso-phosphorus fertilizers as nutrient sources on carp polyculture in Bangladesh

A 120 day long experiment was conducted to find out the effects of cow manure with urea and triple super phosphate (CUT), poultry manure with urea and triple super phosphate (PUT) and cow manure with poultry manure (CP) having similar quantities of nitrogen and phosphorus on pond productivity and fish yield. The stocking fish were rohu (Labeo rohita), catla ( Catla catla) and mrigal ( Cirrhinus mrigala) in each treatment pond at the rate of 10000/ha. All ponds were fertilized fortnightly at the rate of 4000 kg/ha cow manure with 62 kg/ha urea and 65 kg/ha TSP, 2700 kg/ha poultry manure with 62 kg/ha urea and 16 kg/ha TSP, and 4000kg/ha cow manure with 2700 kg/ha poultry manure for the treatment CUT, PUT and CP respectively. Each treatment contained an iso-nitrogen and iso-phosphorus of 56 kg and 46 kg respectively. Though the physico-chemical parameters were more or less similar in all ponds, the chlorophyll-a content and abundance of total plankton were significantly higher (P< 0.05) in the ponds receiving the fertilizer treatment of PUT than those of other treatments. Final growth as well as per unit production of fish of treatment PUT (1773 kg/ha) was significantly higher (P< 0.05) than that of treatment CP (1528 kg/ha) followed by that of treatment CUT (1336 kg/ha). The over all results showed that poultry manure with urea and triple super phosphate proved to be superior to cow manure with urea and triple super phosphate, and poultry manure with cow manure, even when nitrogen and phosphorus content was similar, in carp polyculture system under prevailing conditions.

Alternative phosphorus sources for formulated fish feed

An experiment was conducted to evaluate the possibility of using inorganic fertilizer triple super phosphate (TSP), inorganic fertilizer 16:20 (a 16:20 grade fertilizer contains 16 percent N and 20 percent P20 5), rice-bran and duck-manure as phosphorus sources in formulated fish feed for Nile tilapia ( Oreochromis niloticus). Experiment was conducted for a period of 2 months in net-cages suspended in fertilized earthen ponds and all male sex-reversed Nile tilapia (9.39- 10.37 g) were used in the experiment. Seven treatments including one non-feed treatment were used in this experiment. Treatment 1 (non-feed), treatment 2 (-P) where fish fed with phosphorus non-supplemented diet acted as control 1 and treatment 3, 4, 5, 6 and 7 where fish fed with 3% di-calcium phosphate (DCP), 3% triple supper phosphate (TSP), 7% 16:20 inorganic fertilizer, 30% rice-bran and 30% duck-manure supplemented diet, respectively. Results showed that the TSP and 16:20 grade inorganic fertilizer supplementation in diets as phosphorus sources were equivalent to DCP (Di-calcium phosphate) supplementation in terms of growth performance, feed utilization efficiency and final body composition of Nile tilapia. Ricebran and duck-manure were not found as good phosphorus sources.

Influences of dietary lipid and phosphorus levels on retention and excretion of phosphorus and nitrogen in fingerlings red sea bream, Pagrus major

A laboratory based 2x3 factorial experiment was conducted for 12 weeks to investigate the influences of dietary lipid and phosphorus (P) levels on retention and excretion of phosphorus and nitrogen (N) in fingerling red sea bream. Two levels of lipid (210 and 260 g/kg) and three levels of phosphorus (17, 14 and 12 g/kgˉ¹) in the dry diets were tested. Duplicate groups of 25 red sea bream (average weight 3.74±0.07 g) per 60L glass tank were fed experimental diets three times a day near to satiation level at 22 to 28°C water temperature. A reduction in dietary fish meal from 500 to 300 g/kg dry diet, corresponding to a supplementation in both dietary lipid and P resulted in significant increase in both P and N retention which resulted in the reduction of their excretion by red sea bream. The overall results of the present study demonstrated that both lipid and phosphorus supplementation are necessary for developing less-polluting feed which in turn, reduce fish meal level in the diet of fingerling red sea bream. Further studies in this regard with different size and age groups of red sea bream are warranted.

The effect of organic matter accumulation on phosphorus release in sediment of Chinese shallow lakes

The effects of organic matter in sediment on phosphorus release were studied by field investigations in eight Chinese shallow freshwater lakes with different trophic status and a laboratory experiment. The sediment organic matter content paralleled the trophic status, ranging from 6.1 to 173.0 g kg(-1) (dry weight), with the mean value of 63.1 g kg(-1) (dry weight). It was positively proportional to Soluble reactive phosphorus concentration in the interstitial water in a form of exponential function, but inversely related to the sediment Fe/P ratio. The sediment alkaline phosphatase activity was significantly related not only to the organic matter content (r = 0.829, P < 0.01, n = 120), but also to the soluble reactive phosphorus concentration in interstitial water (r = 0.454, P < 0.01, n = 42). In the laboratory experiment, the addition of organic matter (dry materials of an aquatic macrophyte) into the sediment significantly enhanced alkaline phosphatase activity and soluble reactive phosphorus release. However, in the treatment with organic matter added and aeration, this release was generally prevented in spite of an increase in APA. Hence, sediment organic matter can effectively accelerate phosphorus release by enzymatic hydrolysis and anaerobic desorption. The latter mechanism seems to be more important.

Mitigation of lake eutrophication: Loosen nitrogen control and focus on phosphorus abatement

w Traditionally, nitrogen control is generally considered an important component of reducing lake eutrophication and cyanobacteria blooms. However, this viewpoint is refuted recently by researchers in China and North America. In the present paper, the traditional viewpoint of nitrogen control is pointed out to lack a scientific basis: the N/P hypothesis is just a subjective assumption; bottle bioassay experiments fail to simulate the natural process of nitrogen fixation. Our multi-year comparative research in more than 40 Yangtze lakes indicates that phosphorus is the key factor determining phytoplankton growth regardless of nitrogen concentrations and that total phytoplankton biomass is determined by total phosphorus and not by total nitrogen concentrations. These results imply that, in the field, nitrogen control will not decrease phytoplankton biomass. This finding is supported by a long-term whole-lake experiment from North America. These outcomes can be generalized in terms that a reduction in nitrogen loading may not decrease the biomass of total phytoplankton as it can stimulate blooms of nitrogen-fixing cyanobacteria. To mitigate eutrophication, it is not nitrogen but phosphorus that should be reduced, unless nitrogen concentrations are too high to induce direct toxic impacts on human beings or other organisms. Finally, details are provided on how to reduce controls on nitrogen and how to mitigate eutrophication. (C) 2009 National Natural Science Foundation of China and Chinese Academy of Sciences. Published by Elsevier Limited and Science in China Press. All rights reserved.

Phosphorus in suspended matter and sediments of a hypertrophic lake. A case study: Lake Dianchi, China

From June 2004 to December 2004, Lake Dianchi, which had large scale of cyanobacterial blooms was investigated in order to study P-fractionation in the suspended matter and the sediment. The investigation improves our understanding of phosphorus in Lake Dianchi and the relationship between phosphorus and cyanobacterial blooms. It contributes to the available literature on the behavior of P in hypertrophic lakes. The distribution of P-fractions in Lake Dianchi was not uniform from northwest to south, but was closely related to the trophic status of the whole lake. The concentrations of total phosphorus, labile P (NH4Cl-P), Organic P (NaOH-NRP) and loss on ignition in suspended matter were positively correlated with the strength of cyanobacterial blooms. Total phosphorus in suspended matter was relatively stable for almost half an year and closely related to Chl. a concentration. The main content of organic phosphorus is in the cyanobacterial blooms. The concentrations of phosphorus bound to metal oxides and carbonates (NaOH-SRP and HCl-P) in sediment were similar to NaOH-SRP and HCl-P in the corresponding suspended matter. The latter two forms of P in suspended matter were not affected by cyanobacterial blooms, indicating that the inorganic phosphorus is derived from the sediment after resuspension from the sediment due to wind and wave action. The contribution of the different P-fractions to TP in sediment and in suspended matter indicates that NH4Cl-P in the suspended matter is an important buffer for maintaining dissolved phosphorus in water.