Colorimetric Chemical Differentiation and Detection of Phosphorus in Eutrophic and High Particulate Waters: Advantages of a New Monitoring Approach

Phosphorus (P) is a key factor forcing eutrophication in limnic and marine systems, and all monitoring programs for water quality accordingly include P determinations. However, traditional monitoring does not allow an analysis of the different components involved in the P cycle taking place in the water column. Nonetheless, the implementation of measures addressing eutrophication requires a full understanding of the processes involved in the transformation and transport of P, in all its chemical forms. In this study, the P categories present in a river and its estuary in northern Germany, which discharge into the Baltic Sea, were characterized. Using the molybdenum blue method we found that the classification of P into the traditional fractions (DIP, DOP, POP) applied in the ocean cannot be applied to turbid waters such as rivers because interferences between the fractions seems to occur. Therefore a new nomenclature has been introduced. In addition to total phosphorus (TP) and dissolved molybdate-reactive phosphorus (DRP; previously referred to as inorganic phosphorus), dissolved non-molybdate-reactive phosphorus (DNP), particulate molybdatereactive phosphorus (PRP), and particulate non-molybdate-reactive phosphorus (PNP) were distinguished. The high spatial and temporal variations in the proportions of these forms with respect to the TP concentration well-demonstrate the complexity of the P cycle and the involved P fractions and emphasize the need for expanded monitoring approach. The potential of eutrophication could be underestimated if not all P categories were considered. With the new operational nomenclature the common and standardized molybdenum blue reaction could be used to implement the analysis of various P components into regular monitoring programs

Concentrations and Uptake of Dissolved Organic Phosphorus Compounds in the Baltic Sea

The dissolved organic phosphorus (DOP) pool in marine waters contains a variety of different compounds. Knowledge of the distribution and utilization of DOP by phyto- and bacterioplankton is limited, but critical to our understanding of the marine phosphorus cycle. In the Baltic Sea, detailed information about the composition of DOP and its turnover is lacking. This study reports the concentrations and uptake rates of DOP compounds, namely, adenosine triphosphate (dATP), deoxyribonucleic acid (dDNA), and phospholipids (dPL), in the Baltic Proper and in Finnish coastal waters in the summers of 2011 and 2012. Both areas differed in their dissolved inorganic phosphorus (DIP) concentrations (0.16 and 0.02–0.04 μM), in the C:P (123–178) and N:P (18–27) ratios, and in abundances of filamentous cyanobacteria and of autotrophic and heterotrophic picoplankton. The mean concentrations of dATP-P, dDNA-P, and dPL-P were 4.3–6.4, 0.05–0.12, and 1.9–6.8 nM, respectively, together contributing between 2.4 and 5.2% of the total DOP concentration. The concentrations of the compounds varied between and within the investigated regions and the distribution patterns of the individual components are not linked to each other. DIP was taken up at rates of 10.1–380.8 nM d-1. dATP-P and dDNA-P were consumed simultaneously with DIP at rates of 6.9–24.1 and 0.09–0.19 nM d-1, respectively, with the main proportion taken up by the size fraction <3 μm and with DIP to be the dominant source. Groups of hydrographical and biological parameters were identified in the multiple regression analysis to impact the concentrations and uptake rates. It points to the complexity of the regulation. Our results indicate that the investigated DOP compounds, particularly dATP-P, can make significant contributions to the P nutrition of microorganisms and their use seems to be not intertwined. Therefore, more detailed knowledge of all DOP components including variation of concentrations and the utilization is required to understand the roles of DOP in marine ecosystems

Effects of CO2 perturbation on phosphorus pool sizes and uptake in a mesocosm experiment during a low productive summer season in the northern Baltic Sea

Studies investigating the effect of increasing CO2 levels on the phosphorus cycle in natural waters are lacking although phosphorus often controls phytoplankton development in many aquatic systems. The aim of our study was to analyse effects of elevated CO2 levels on phosphorus pool sizes and uptake. The phosphorus dynamic was followed in a CO2-manipulation mesocosm experiment in the Storfjarden (western Gulf of Finland, Baltic Sea) in summer 2012 and was also studied in the surrounding fjord water. In all mesocosms as well as in surface waters of Storfjarden, dissolved organic phosphorus (DOP) concentrations of 0.26aEuro-+/- aEuro-0.03 and 0.23aEuro-+/- aEuro-0.04aEuro-A mu molaEuro-L-1, respectively, formed the main fraction of the total P-pool (TP), whereas phosphate (PO4) constituted the lowest fraction with mean concentration of 0.15aEuro-A +/- aEuro-0.02 in the mesocosms and 0.17aEuro-A +/- aEuro-0.07aEuro-A mu molaEuro-L-1 in the fjord. Transformation of PO4 into DOP appeared to be the main pathway of PO4 turnover. About 82aEuro-% of PO4 was converted into DOP whereby only 18aEuro-% of PO4 was transformed into particulate phosphorus (PP). PO4 uptake rates measured in the mesocosms ranged between 0.6 and 3.9aEuro-nmolaEuro-L(-1)aEuro-h(-1). About 86aEuro-% of them was realized by the size fraction aEuro-1000aEuro-A mu atm during periods when phytoplankton biomass increased. In addition, we found significant relationships (e.g., between PP and Chl a) in the untreated mesocosms which were not observed under high fCO(2) conditions. Consequently, it can be hypothesized that the relationship between PP formation and phytoplankton growth changed with CO2 elevation. It can be deduced from the results, that visible effects of CO2 on P pools are coupled to phytoplankton growth when the transformation of PO4 into POP was stimulated. The transformation of PO4 into DOP on the other hand does not seem to be affected. Additionally, there were some indications that cellular mechanisms of P regulation might be modified under CO2 elevation changing the relationship between cellular constituents.Peer reviewe

2008 Inter-laboratory Comparison Study of a Reference Material for Nutrients in Seawater

Autoclaved natural seawater collected in the North Pacific Ocean was used as a reference material for nutrients in seawater (RMNS) during an inter-laboratory comparison (I/C) study conducted in 2008. This study was a follow-up to previous studies conducted in 2003 and 2006. A set of six samples was distributed to each of 58 laboratories in 15 countries around the globe, and results were returned by 54 of those laboratories (15 countries). The homogeneities of samples used in the 2008 I/C study, based on analyses for three determinants, were improved compared to those of samples used in the 2003 and 2006 I/C studies. Results of these I/C studies indicate that most of the participating laboratories have an analytical technique for nutrients that is sufficient to provide data of high comparability. The differences between reported concentrations from the same laboratories in the 2006 and 2008 I/C studies for the same batch of RMNS indicate that most of the laboratories have been maintaining internal comparability for two years. Thus, with the current high level of performance in the participating laboratories, the use of a common reference material and the adaptation of an internationally accepted nutrient scale system would increase comparability among laboratories worldwide, and the use of a certified reference material would establish traceability. In the 2008 I/C study we observed a problem of non-linearity of the instruments of the participating laboratories similar to that observed among the laboratories in the 2006 I/C study. This problem of non-linearity should be investigated and discussed to improve comparability for the full range of nutrient concentrations. For silicate comparability in particular, we see relatively larger consensus standard deviations than those for nitrate and phosphate

KOSMOS Finland 2012 mesocosm study: Phosphorus pool sizes and uptake

Studies investigating the effect of increasing CO2 levels on the phosphorus cycle in natural waters are lacking although phosphorus often controls phytoplankton development in many aquatic systems. The aim of our study was to analyse effects of elevated CO2 levels on phosphorus pool sizes and uptake. The phosphorus dynamic was followed in a CO2-manipulation mesocosm experiment in the Storfjärden (western Gulf of Finland, Baltic Sea) in summer 2012 and was also studied in the surrounding fjord water. In all mesocosms as well as in surface waters of Storfjärden, dissolved organic phosphorus (DOP) concentrations of 0.26 ± 0.03 and 0.23 ± 0.04 µmol/l, respectively, formed the main fraction of the total P-pool (TP), whereas phosphate (PO4) constituted the lowest fraction with mean concentration of 0.15 ± 0.02 in the mesocosms and 0.17 ± 0.07 µmol/l in the fjord. Transformation of PO4 into DOP appeared to be the main pathway of PO4 turnover. About 82% of PO4 was converted into DOP whereby only 18% of PO4 was transformed into particulate phosphorus (PP). PO4 uptake rates measured in the mesocosms ranged between 0.6 and 3.9 nmol/l/h. About 86% of them was realized by the size fraction 1000 µatm during periods when phytoplankton biomass increased. In addition, we found significant relationships (e.g., between PP and Chl a) in the untreated mesocosms which were not observed under high fCO2 conditions. Consequently, it can be hypothesized that the relationship between PP formation and phytoplankton growth changed with CO2 elevation. It can be deduced from the results, that visible effects of CO2 on P pools are coupled to phytoplankton growth when the transformation of PO4 into POP was stimulated. The transformation of PO4 into DOP on the other hand does not seem to be affected. Additionally, there were some indications that cellular mechanisms of P regulation might be modified under CO2 elevation changing the relationship between cellular constituents

The relationship between dissolved carbohydrates and carbohydrate-degrading enzymes in the salinity gradient of the Pomeranian Bight (southern Baltic)

From 1994 to 1996 changes in the concentrations of dissolved mono-(MCHO) and total dissolved polysaccharides (TCHO) as well as the activities of carbohydrate-degrading enzymes &#945;- and &#946;-glucosidase, glucosaminidase) were investigated during mixing of water from the River Odra and the open Pomeranian Bight. This study addresses the question of whether their distribution was a result of physical dilution alone or if biological interactions were detectable. <br>&nbsp;&nbsp;&nbsp;&nbsp;Within the salinity gradient, ranging from 1.9 to 7.8 PSU, TCHO declined from 13.2 µmol l^-1 near the &#346;wina mouth to 2.8 µmol l^-1 after mixing. Concentrations of MCHO decreased from 3.4 µmol l^-1 to 1.1 µmol l^-1 but its distribution pattern varied more between summer and autumn than that of TCHO. The hydrolysis rate (Hr) by glucosidase and glucosaminidase activities was reduced from 13.9% h^-1 to 0.3% h^-1 and 9.9% h^-1 to 0.2% h^-1, respectively, and correlated with the uptake rate of glucose (To) by bacteria. In summer, the To/Hr ratio increased from about 1.2 to 29.4, mainly because of stronger decreases in Hr than in To. It was shown that the relationship between enzymatic release and uptake of carbohydrates influences the concentration of dissolved carbohydrates within the salinity gradient. Most probably, the decrease in carbohydrate-degrading enzymes is the result of reduced substrate stimulation and the lower number of particle-associated bacteria

Response of Nodularia spumigena to pCO2 - Part 3: Turnover of phosphorus compounds

Diazotrophic cyanobacteria often form extensive summer blooms in the Baltic Sea driving their environment into phosphate limitation. One of the main species is the heterocystous cyanobacterium Nodularia spumigena. N. spumigena exhibits accelerated uptake of phosphate through the release of the exoenzyme alkaline phosphatase that also serves as an indicator of the hydrolysis of dissolved organic phosphorus (DOP). The present study investigated the utilization of DOP and its compounds (e.g. ATP) by N. spumigena during growth under varying CO2 concentrations, in order to estimate potential consequences of ocean acidification on the cell's supply with phosphorus. Cell growth, phosphorus pool fractions, and four DOP-compounds (ATP, DNA, RNA, and phospholipids) were determined in three set-ups with different CO2 concentrations (341, 399, and 508 µatm) during a 15-day batch experiment. The results showed rapid depletion of dissolved inorganic phosphorus (DIP) in all pCO2 treatments while DOP utilization increased with elevated pCO2, in parallel with the growth stimulation of N. spumigena. During the growth phase, DOP uptake was enhanced by a factor of 1.32 at 399 µatm and of 2.25 at 508 µatm compared to the lowest pCO2 concentration. Among the measured DOP compounds, none was found to accumulate preferentially during the incubation or in response to a specific pCO2 treatment. However, at the beginning 61.9 ± 4.3% of the DOP were not characterized but comprised the most highly utilized fraction. This is demonstrated by the decrement of this fraction to 27.4 ± 9.9% of total DOP during the growth phase, especially in response to the medium and high pCO2 treatment. Our results indicate a stimulated growth of diazotrophic cyanobacteria at increasing CO2 concentrations that is accompanied by increasing utilization of DOP as an alternative P source

Phosphorus input by upwelling in the eastern Gotland Basin (Baltic Sea) in summer and its effects on filamentous cyanobacteria

In July 2007, phosphorus input by an upwelling event along the east coast of Gotland Island and the response of filamentous cyanobacteria were studied to determine whether introduced phosphorus can intensify cyanobacterial bloom formation in the eastern Gotland Basin. Surface temperature, nutrient concentrations, phytoplankton biomass and its stoichiometry, as well as phosphate uptake rates were determined in two transects between the coasts of Gotland and Latvia and in a short grid offshore of Gotland. In the upwelling area, surface temperatures of 11–12 °C and average dissolved inorganic phosphorus (DIP) concentrations of 0.26 μM were measured. Outside the upwelling, surface temperatures were higher (15.5–16.6 °C) and DIP supplies in the upper 10 m layer were exhausted. Nitrite and nitrate concentrations (0.01–0.22 μM) were very low within and outside the upwelling region. Abundances of filamentous cyanobacteria were highly reduced in the upwelling area, accounting for only 1.4–6.0% of the total phytoplankton biomass, in contrast to 18–20% outside the upwelling. The C:P ratio of filamentous cyanobacteria varied between 32.8 and 310 in the upwelling region, most likely due to the introduction of phosphorus-depleted organisms into the upwelling water. These organisms accumulate DIP in upwelling water and have lower C:P ratios as long as they remain in DIP-rich water. Thus, diazotrophic cyanobacteria benefit from phosphorus input directly in the upwelling region. Outside the upwelling region, the C:P ratios of filamentous cyanobacteria varied widely, between 240 and 463, whereas those of particulate material in the water ranged only between 96 and 224. To reduce their C:P ratio from 300 to 35, cyanobacteria in the upwelling region had to take up 0.05 mmol m−3 DIP, which is about 20% of the available DIP. Thus, a larger biomass of filamentous cyanobacteria may be able to benefit from a given DIP input. As determined from the DIP uptake rates measured in upwelling cells, the time needed to reduce the C:P ratio from 300 to 35 was too long to explain the huge bloom formations that typically occur in summer. However, phosphorus uptake rates increased significantly with increasing C:P ratios, allowing phosphorus accumulation within 4–5 days, a span of time suitable for bloom formation in July and August

Variations in the elemental ratio of organic matter in the central Baltic Sea: Part I—Linking primary production to remineralization

Highlights: • Elemental C:N:P variations of organic matter are simulated at monitoring site BY15. • No N2 fixation needed to explain observed PO4PO4 and pCO2pCO2 levels after spring bloom. • Model features relevance of DOP production and remineralization for N2 fixation. • Model estimates of annual N2 fixation are View the MathML source297±24mmolNm-2a-1. • Model estimates of annual total production are View the MathML source14.16±0.71molCm-2a-1. Abstract: For most marine ecosystems the growth of diazotrophic cyanobacteria and the associated amount of nitrogen fixation are regulated by the availability of phosphorus. The intensity of summer blooms of nitrogen (N2) fixing algae in the Baltic Sea is assumed to be determinable from a surplus of dissolved inorganic phosphorus (DIP) that remains after the spring bloom has ended. But this surplus DIP concentration is observed to continuously decrease at times when no appreciable nitrogen fixation is measured. This peculiarity is currently discussed and has afforded different model interpretations for the Baltic Sea. In our study we propose a dynamical model solution that explains these observations with variations of the elemental carbon-to-nitrogen-to-phosphorus (C:N:P) ratio during distinct periods of organic matter production and remineralization. The biogeochemical model resolves seasonal C, N and P fluxes with depth at the Baltic Sea monitoring site BY15, based on three assumptions: (1) DIP is utilized by algae though not needed for immediate growth, (2) the uptake of dissolved inorganic nitrogen (DIN) is hampered when the algae׳s phosphorus (P) quota is low, and (3) carbon assimilation continues at times of nutrient depletion. Model results describe observed temporal variations of DIN, DIP and chlorophyll-a concentrations along with partial pressure of carbon dioxide (pCO2)(pCO2). In contrast to other model studies, our solution does not require N2 fixation to occur shortly after the spring bloom to explain DIP drawdown and pCO2pCO2 levels. Model estimates of annual N2 fixation are View the MathML source297±24mmolNm-2a-1. Estimates of total production are View the MathML source14200±700mmolCm-2a-1, View the MathML source1400±70mmolNm-2a-1, and View the MathML source114±5mmolPm-2a-1 for the upper 50 m. The models C, N and P fluxes disclose preferential remineralization of P and of organic N that was introduced via N2 fixation. Our results are in support of the idea that P uptake by phytoplankton during the spring bloom contributes to the consecutive availability of labile dissolved organic phosphorus (LDOP). The LDOP is retained within upper layers and its remineralization affects algal growth in summer, during periods of noticeable N2 fixation