Comparison of the acid-base responses to CO2 and acidification in Japanese flounder (Paralichthys olivaceus)

To investigate whether the biological toxicity of aquatic hypercapnia is due to the direct effects of CO2 or to the effects of acidification of seawater by CO2, the Japanese flounder (Paralichthys olivaceus) was subjected to seawater equilibrated with a gas mixture of air containing 5% CO2 (pH 6.18) or seawater acidified to the same pH with 1 N H2SO4. All the fish died within 72 h in the CO2 exposure group, whereas no mortality occurred in the acid group. Acid-base parameters as well as plasma ion concentrations were severely perturbed in the CO2 exposure group, whereas they were minimally affected in the acid group. These results clearly demonstrate that the mortality in the CO2 group is a direct result of the elevated levels of dissolved CO2 and not to the effects of the reduced water pH.     

The inhibition of marine nitrification by ocean disposal of carbon dioxide

In an attempt to reduce the threat of global warming, it has been proposed that the rise of atmospheric carbon dioxide concentrations be reduced by the ocean disposal of CO2 from the flue gases of fossil fuel-fired power plants. The release of large amounts of CO2 into mid or deep ocean waters will result in large plumes of acidified seawater with pH values ranging from 6 to 8. In an effort to determine whether these CO2-induced pH changes have any effect on marine nitrification processes, surficial (euphotic zone) and deep (aphotic zone) seawater samples were sparged with CO2 for varying time durations to achieve a specified pH reduction, and the rate of microbial ammonia oxidation was measured spectrophotometrically as a function of pH using an inhibitor technique. For both seawater samples taken from either the euphotic or aphotic zone, the nitrification rates dropped drastically with decreasing pH. Relative to nitrification rates in the original seawater at pH 8, nitrification rates were reduced by ca. 50% at pH 7 and more than 90% at pH 6.5. Nitrification was essentially completely inhibited at pH 6. These findings suggest that the disposal of CO2 into mid or deep oceans will most likely result in a drastic reduction of ammonia oxidation rates within the pH plume and the concomitant accumulation of ammonia instead of nitrate. It is unlikely that ammonia will reach the high concentration levels at which marine aquatic organisms are known to be negatively affected. However, if the ammonia-rich seawater from inside the pH plume is upwelled into the euphotic zone, it is likely that changes in phytoplankton abundance and community structure will occur. Finally, the large-scale inhibition of nitrification and the subsequent reduction of nitrite and nitrate concentrations could also result in a decrease of denitrification rates which, in turn, could lead to the buildup of nitrogen and unpredictable eutrophication phenomena. Clearly, more research on the environmental effects of ocean disposal of CO2 is needed to determine whether the potential costs related to marine ecosystem disturbance and disruption can be justified in terms of the perceived benefits that may be achieved by temporarily delaying global warming.     

Effects of high CO2 seawater on the copepod (Acartia tsuensis) through all life stages and subsequent generations

We studied the effects of exposure to seawater equilibrated with CO(2)-enriched air (CO(2) 2380 ppm) from eggs to maturity and over two subsequent generations on the copepod Acartia tsuensis. Compared to the control (CO(2) 380 ppm), high CO(2) exposure through all life stages of the 1st generation copepods did not significantly affect survival, body size or developmental speed. Egg production and hatching rates were also not significantly different between the initial generation of females exposed to high CO(2) and the 1st and 2nd generation females developed from eggs to maturity in high CO(2). Thus, the copepods appear more tolerant to increased CO(2) than other marine organisms previously investigated for CO(2) tolerance (i.e., sea urchins and bivalves). However, the crucial importance of copepods in marine ecosystems requires thorough evaluation of the overall impacts of marine environmental changes predicted to occur with increased CO(2) concentrations, i.e., increased temperature, enhanced UV irradiation, and changes in the community structure and nutritional value of phytoplankton.     

Effects of anthropogenic seawater acidification on acid-base balance in the sea urchin Psammechinus miliaris

The purple-tipped sea urchin, Psammechinus miliaris, was exposed to artificially acidified seawater treatments (pH(w) 6.16, 6.63 or 7.44) over a period of 8 days. Urchin mortality of 100% was observed at pH(w) 6.16 after 7 days and coincided with a pronounced hypercapnia in the coelomic fluid producing an irrecoverable acidosis. Coelomic fluid acid-base measures showed that an accumulation of CO(2) and a significant reduction in pH occurred in all treatments compared with controls. Bicarbonate buffering was employed in each case, reducing the resultant acidosis, but compensation was incomplete even under moderate environmental hypercapnia. Significant test dissolution was inferred from observable increases in the Mg(2+) concentration of the coelomic fluid under all pH treatments. We show that a chronic reduction of surface water pH to below 7.5 would be severely detrimental to the acid-base balance of this predominantly intertidal species; despite its ability to tolerate fluctuations in pCO(2) and pH in the rock pool environment. The absence of respiratory pigment (or any substantial protein in the coelomic fluid), a poor capacity for ionic regulation and dependency on a magnesium calcite test, make echinoids particularly vulnerable to anthropogenic acidification. Geological sequestration leaks may result in dramatic localised pH reductions, e.g. pH 5.8. P. miliaris is intolerant of pH 6.16 seawater and significant mortality is seen at pH 6.63.     

Detrimental effects of reduced seawater pH on the early development of the Pacific abalone

The hatching process of the Pacific abalone Haliotis discus hannai was prolonged at a pH of 7.6 and pH 7.3, and the embryonic developmental success was reduced. The hatching rate at pH 7.3 was significantly (10.8%) lower than that of the control (pH 8.2). The malformation rates at pH 7.9 and pH 8.2 were less than 20% but were 53.8% and 77.3% at pH 7.6 and pH 7.3, respectively. When newly hatched larvae were incubated for 48 h at pH 7.3, only 2.7% of the larvae settled, while more than 70% of the larvae completed settlement in the other three pH treatments. However, most 24 h old larvae could complete metamorphosis in all four pH treatments. Overall, a 0.3-unit reduction in water pH will produce no negative effect on the early development of the Pacific abalone, but further reduction in pH to the values predicted for seawater by the end of this century will have strong detrimental effects.     

A marine secondary producer respires and feeds more in a high CO2 ocean

Climate change mediates marine chemical and physical environments and therefore influences marine organisms. While increasing atmospheric CO(2) level and associated ocean acidification has been predicted to stimulate marine primary productivity and may affect community structure, the processes that impact food chain and biological CO(2) pump are less documented. We hypothesized that copepods, as the secondary marine producer, may respond to future changes in seawater carbonate chemistry associated with ocean acidification due to increasing atmospheric CO(2) concentration. Here, we show that the copepod, Centropages tenuiremis, was able to perceive the chemical changes in seawater induced under elevated CO(2) concentration (>1700 μatm, pH<7.60) with avoidance strategy. The copepod's respiration increased at the elevated CO(2) (1000 μatm), associated acidity (pH 7.83) and its feeding rates also increased correspondingly, except for the initial acclimating period, when it fed less. Our results imply that marine secondary producers increase their respiration and feeding rate in response to ocean acidification to balance the energy cost against increased acidity and CO(2) concentration.     

Benthic foraminifera show some resilience to ocean acidification in the northern Gulf of California,Mexico

Extensive CO₂ vents have been discovered in the Wagner Basin, northern Gulf of California, where they create large areas with lowered seawater pH. Such areas are suitable for investigations of long-term biological effects of ocean acidification and effects of CO₂ leakage from subsea carbon capture storage. Here, we show responses of benthic foraminifera to seawater pH gradients at 74–207 m water depth. Living (rose Bengal stained) benthic foraminifera included Nonionella basispinata, Epistominella bradyana and Bulimina marginata. Studies on foraminifera at CO₂ vents in the Mediterranean and off Papua New Guinea have shown dramatic long-term effects of acidified seawater. We found living calcareous benthic foraminifera in low pH conditions in the northern Gulf of California, although there was an impoverished species assemblage and evidence of post-mortem test dissolution.     

Effects of antifouling paint components (TBTO,copper and triazine) on the early development of embryos in cod (Gadus morhua L.)

To assess the risk of antifoulant use to the commercially important cod (Gadus morhua L.), fertilised cod eggs were exposed to triazine, copper and TBTO singly or combined in laboratory tests with running seawater. At the highest tested concentrations (11.5 microg Cu l(-1); 5 microg TBTO l(-1)) larval mortality was increased. The highest concentration of triazine (40 microg l(-1)) did not cause any significant mortality. Fertilised eggs that had been exposed to all the three chemicals singly for five days showed a higher buoyancy than the controls. No synergistic or antagonistic effects were indicated. Embryos/larvae exposed to 0.004-0.8 microg TBTO l(-1) did not show any changed respiration compared to the controls after hatching. It is concluded that existing known field concentrations of the three antifoulants are hardly expected to cause detectable effects on fish embryonic/larval development.     

Acute toxicity of temporally varying seawater CO2 conditions on juveniles of Japanese sillago (Sillago japonica)

CO(2) ocean storage by which liquefied CO(2) is injected into the deep-sea to mitigate the climate change would increase the CO(2) concentrations of the surrounding seawater. The biological impacts of such dynamic CO(2) environments are, however, unknown. We examined the acute toxicity of temporally changing seawater CO(2) concentrations on juveniles of Sillago japonica. Step-wise increases in ambient CO(2) to fCO(2) (fractional CO(2) concentration of the gas mixture bubbled into seawater) levels of 7% and 9% resulted in mortalities of 0.15 and 0.40-0.67 after 18 h, respectively. In contrast, one-step increases to these CO(2) levels killed all fish within 15 min. Further, a sudden drop of fCO(2) from 9-10% CO(2) to normocapnia (0.038%) killed all the surviving fish within a few minutes. These results demonstrate that impacts of ocean CO(2) storage need to be examined under conditions mimicking the dynamic changes in CO(2) levels expected to occur by the CO(2) injection procedure.     

Effects of raised CO2 concentration on the egg production rate and early development of two marine copepods (Acartia steueri and Acartia erythraea)

Direct injection of CO(2) into the deep ocean is receiving increasing attention as a way to mitigate increasing atmospheric CO(2) concentration. To assess the potential impact of the environmental change associated with CO(2) sequestration in the ocean, we studied the lethal and sub-lethal effects of raised CO(2) concentration in seawater on adult and early stage embryos of marine planktonic copepods. We found that the reproduction rate and larval development of copepods are very sensitive to increased CO(2) concentration. The hatching rate tended to decrease, and nauplius mortality rate to increase, with increased CO(2) concentration. These results suggest that the marine copepod community will be negatively affected by the disposal of CO(2). This could decrease on the carbon export flux to the deep ocean and change the biological pump. Clearly, further studies are needed to determine whether ocean CO(2) injection is an acceptable strategy to reduce anthropogenic CO(2).     

Variation and temporal patterns in the composition of the surface ichthyoplankton in the southern Bay of Biscay (W. Atlantic)

From September 2000 to December 2006, surface plankton samples were collected on a monthly basis, from a station located in the southern Bay of Biscay (43°37N; 1°43W France), near the deep Capbreton canyon. In this paper, the results for the ichthyoplankton assemblage are presented. Among the 62 taxa recorded, only 35 were present in the larval stage whilst only 10 were represented by their eggs. Taxa represented by both stages (eggs+larvae; N=17) had the highest abundance. The presence in the surface plankton assemblage of species, at either or both stage, is interpreted within the context of the bathymetric distribution of species. The maxima in abundance and diversity occurred in February–March, for eggs, and May–June, for larvae. This 3-month time-lag between the stages is proposed to be related to the timing of egg spawning and larval recruitment to the pelagic environment. Mean egg abundances (82.4±29.8 eggs/10 m²) were 10-fold higher than the larval abundances (7.1±1.8 larvae/10 m²). Despite pronounced monthly variability, no statistically significant decrease in either egg or larvae abundance was observed during this 6-year study period. Compared with previous published studies, this study shows that the peak in ichthyoplankton diversity occurred two months earlier. In addition, the spawning period occurred over the whole year, even during autumn and winter. Using ordination techniques, the annual sequence appearance of taxa are described at the study site: Gadiforms, Ammodytidae and Pleuronectiforms were present during the winter whilst Sparidae, Blennidae, Labridae and Gobiidae, formed the summer group. Only three species, European anchovy Engraulis encrasicolus, European pilchard Sardina pilchardus and Atlantic horse mackerel Trachurus trachurus were recorded throughout the year.     

High mortality of the larvae of the common mussel at low concentrations of tributyltin

Laboratory reared veliger larvae of the common mussel Mytilus edulis were grown for 15 days in filtered seawater containing 10 μg l^?1, 1 μg l^?1 and 0.1 μg l^?1 tributyltin oxide (TBTO), the biocide in recent antifouling paint formulations. No larvae survived longer than 5 days in 10 μg l^?1 TBTO or 10 days in 1 μg l^?1 TBTO. Approximately half the larvae subjected to 0.1 μg l^?1 TBTO were dead by day 15 (approximate 15 day LC₅₀ = 0.1 μg l^?1 TBTO) and most of the surviving larvae were moribund and had grown significantly more slowly than controls. It is noted that current recorded levels of TBT at several UK estuarine sites are higher than 0.1 μg l^?1 and there has therefore probably been a high mortality of mussel larvae at such sites. The results are discussed in relation to the reported short-term toxicity of organotins to other marine organisms and the value of carrying out longer-term tests at low levels of TBT is pointed out.     

U,U andTh in seawater

We have developed techniques to determine²³⁸U,²³⁴U and²³²Th concentrations in seawater by isotope dilution mass spectrometry. U measurements are made using a²³³U²³⁶U double spike to correct for instrumental fractionation. Measurements on uranium standards demonstrate that²³⁴U/²³⁸U ratios can be measured accurately and reproducibly.²³⁴U/²³⁸U can be measured routinely to ± 5‰ (2σ) for a sample of 5 × 10⁹ atoms of²³⁴U (3 × 10⁻⁸ g of total U, 10 ml of seawater). Data acquisition time is 1 hour. The small sample size, high precision and short data acquisition time are superior to-counting techniques.²³⁸U is measured to ± 2‰ (2σ) for a sample of 8 × 10¹² atoms of²³⁸U ( 3 × 10⁻⁹ g of U, 1 ml of seawater).²³²Th is measured to ± 20‰ with 3 × 10¹¹²³²Th atoms (10⁻¹⁰ g²³²Th, 1 1 of seawater). This small sample size will greatly facilitate investigation of the²³²Th concentration in the oceans. Using these techniques, we have measured²³⁸U,²³⁴U and²³²Th in vertical profiles of unfiltered, acidified seawater from the Atlantic and²³⁸U and²³⁴U in vertical profiles from the Pacific. Determinations of²³⁴U/²³⁸U at depths ranging from 0 to 4900 m in the Atlantic (7°44′N, 40°43′W) and the Pacific (14°41′N, 160°01′W) Oceans are the same within experimental error (± 5‰,2σ). The average of these²³⁴U/²³⁸U measurements is 144 ± 2‰ (2σ) higher than the equilibrium ratio of 5.472 × 10⁻⁵. U concentrations, normalized to 35‰ salinity, range from 3.162 to 3.281 ng/g, a range of 3.8%. The average concentration of the Pacific samples (31°4′N, 159°1′W) is 1% higher than that of the Atlantic (7°44′N, 40°43′W and 31°49′N, 64°6′W).²³²Th concentrations from an Atlantic profile range from 0.092 to 0.145 pg/g. The observed constancy of the²³⁴U/²³⁸U ratio is consistent with the predicted range of²³⁴U/²³⁸U using a simple two-☐ model and the residence time of deep water in the ocean determined from¹⁴C. The variation in salinity-normalized U concentrations suggests that U may be much more reactive in the marine environment than previously thought.     

Respiration rate and swimming activity of larvae of two sub-tidal nassariid gastropods under reduced oxygen levels: implications for their distributions in Hong Kong waters

The effects of hypoxia on the larvae of two sub-tidal nassariid gastropods, Nassarius siquijorensis and N. conoidalis were compared so as to understand how the species-specific tolerance to hypoxia might have resulted in changes in the abundance and distribution of these two species in the hypoxic Tolo Harbour, Hong Kong, since the 1980s. Respiration rates of N. siquijorensis and N. conoidalis larvae were reduced at 4.5 mg O2 l(-1), or below, as compared with the normoxic control. Significant reduction in swimming velocity was also observed for 10-day old larvae which were exposed to <2.0 mg O2 l(-1) for N. siquijorensis and <1.0 mg O2 l(-1) for N. conoidalis. The 48 h LC50 values of N. siquijorensis and N. conoidalis larvae were 0.7 and 1.7 mg O2 l(-1), respectively. The results suggested that N. siquijorensis are more tolerant to hypoxia than N. conoidalis.     

Habitat selection by marine larvae in changing chemical environments

The replenishment and persistence of marine species is contingent on dispersing larvae locating suitable habitat and surviving to a reproductive stage. Pelagic larvae rely on environmental cues to make behavioural decisions with chemical information being important for habitat selection at settlement. We explored the sensory world of crustaceans and fishes focusing on the impact anthropogenic alterations (ocean acidification, red soil, pesticide) have on conspecific chemical signals used by larvae for habitat selection. Crustacean (Stenopus hispidus) and fish (Chromis viridis) larvae recognized their conspecifics via chemical signals under control conditions. In the presence of acidified water, red soil or pesticide, the ability of larvae to chemically recognize conspecific cues was altered. Our study highlights that recruitment potential on coral reefs may decrease due to anthropogenic stressors. If so, populations of fishes and crustaceans will continue their rapid decline; larval recruitment will not replace and sustain the adult populations on degraded reefs.     

The impact of atmospheric aerosols on trace metal chemistry in open ocean surface seawater, 1. Aluminum

Significant quantities of aerosol aluminum are transported from continental regions through the atmosphere to the oceans. Enrichments in the concentration of dissolved aluminum in open ocean surface seawater suggest that dissolution of aerosol aluminum is an important source of dissolved aluminum to these waters. Atmospheric aerosols collected at Enewetak Atoll were exposed to seawater and artificial rain water to determine directly the importance of atmospheric deposition as a source of marine dissolved aluminum.The results of these experiments indicate that 8–10% of the aluminum in atmospheric aerosols of crustal origin over the North Pacific is soluble in seawater. Approximately 5–6% dissolves very rapidly ( < 0.6 hr). An additional 3–4% dissolves within 60 hr. This bimodal dissolution of aerosol aluminum of crustal origin suggests that this aluminum is present in two forms. The rapidly dissolving fraction is likely aluminum already weathered from primary minerals, while the more slowly dissolving fraction is probably aluminum from the aluminosilicate matrix.Nearly the same amount of aerosol aluminum dissolved in artificial rain water (pH= 5.5) in 6 hr as dissolved in seawater (pH= 8) in 60 hr. The lower pH appears to not only increase the dissolution rate but may also increase the quantity of aerosol aluminum that dissolves. Dissolved organic matter in seawater appears to have relatively little effect on aerosol aluminum dissolution.Considering measured total aerosol aluminum fluxes, aluminum dissolution of 5–10% would constitute the major source for dissolved aluminum in surface waters of the open North Pacific. The calculated residence time of dissolved aluminum in the upper 100 m of the tropical North Pacific ranges from 2 to 6 years.     

Seasonal and spatial characteristics of seawater and sediment at Youngil Bay, southeast coast of Korea

The seasonal geochemical characteristics of the seawater and sediments and the major factors causing heavy metal contamination were investigated at the Youngil bay and the Hyungsan river estuary in the Southeast Coast of Korea, where a world-scale steel-industry complex (Pohang iron and steel industrial complex, POSCO) is located. The seasonal and spatial distribution characteristics of temperature, dissolved oxygen (DO), pH, and nutrients of the seawater were studied at 45 fixed stations, especially focusing on the river mouth area. Sediments at 27 stations were examined during winter and summer to determine the major controlling factors for the distribution of metals, using correlation matrix and R-mode factor analyses, and to evaluate the pollution status, using the modified geoaccumulation (I(geo)(')) index. Temperatures for the effluent from the POSCO located at the Hyungsan river mouth were 2-3 degrees C higher compared to other sampling areas, due to the thermal discharge from the POSCO. The DO concentration of the surface water at the Pohang old port was as low as 2-4 mg/L. In spring, the DO value at the Hyungsan river mouth was higher than 12 mg/L, by the mass multiplication of phytoplanktons at the river mouth where seawater temperature and nutrients concentrations were relatively high, resulting in the pH value of higher than 8.3. The nitrogen to phosphorus (N/P) ratios at the river mouth were 20-150 times higher compared to other areas, implying that the nitrogen loading into this semi-enclosed bay is significantly higher than phosphorus and the major nitrogen sources are not only the domestic sewage from the city but the industrial wastewater from the POSCO and other steel factories nearby. The phosphorus concentrations at the Pohang old port were shown 3-10 times higher than those at other stations, due to the inflow of pollutants generated from the nearby ships anchoring and the release of phosphate from the bottom sediment. Results from the sediment analysis showed that the major controlling factors for the distribution pattern of each metal are grain size and organic carbon (C(org)) content. Based on the factor analysis, Al, Fe, Cr, Li, and Pb were shown strongly correlated with the mean grain size (Mz), whereas Cd, Cu, Zn, and Sn with the C(org) content. Results from the fractionation of the sedimentary metals into lattice and labile fractions to characterize the mobility of sediment metals showed that the mineral lattice fraction was high in the order of Al=K>Cr>Li>Sr>Fe, while the labile fraction, which might be released to the overlying water, was in the order of Pb>Zn>Cd>Cu>Ca>Sn. Evaluation of the sediment pollution status by applying (I(geo)(')) of 13 metals showed Cd, Cu, and Zn as high as 1-3 range at the old port. Even though the overall marine pollution mainly by the world-class steel industrial complex in this semi-enclosed bay area studied does not currently pose a serious threat, due to the seawater circulation and the large influx of river discharge, the countermeasures to implement the sediments concentrated with heavy metals, especially at the old port with no seawater circulation, are still warranted for this coastal water environment.     

The Effect of Copper on the Settlement Success of Larvae from the Scleractinian Coral Acropora tenuis

This study examined the effect of copper on the settlement success of planula larvae of the reef-building coral Acropora tenuis during 1994 and 1996 at Magnetic Island, Great Barrier Reef. Copper concentrations of 2, 10, 20 μg l⁻¹ did not inhibit larval settlement after 48-h exposure. However, copper concentrations of 42 μg l⁻¹ and 81 μg l⁻¹ significantly reduced settlement success of A. tenuis larvae after 48-h exposure compared with controls using normal seawater. At 200 μg l⁻¹ copper, all larvae died. EC₅₀ values for the effect of copper on A. tenuis larval settlement were calculated from the 1996 results using measured copper concentrations. The 48-h EC₅₀ was 35 μg l⁻¹ with an upper and lower 95% confidence limit of 37 μg l⁻¹ and 32 μg l⁻¹, respectively. The 48-h NOEC value for both experiments was 20 μg l⁻¹ copper. These experiments provide some of the first data on sub-lethal effects of trace metals on tropical marine organisms, and demonstrate that relatively low copper concentrations impair or inhibit settlement of coral larvae.     

Impact of elevated levels of CO2 on animal mediated ecosystem function: The modification of sediment nutrient fluxes by burrowing urchins

A mesocosm experiment was conducted to quantify the relationships between the presence and body size of two burrowing heart urchins (Brissopsis lyrifera and Echinocardium cordatum) and rates of sediment nutrient flux. Furthermore, the impact of seawater acidification on these relationships was determined during this 40-day exposure experiment. Using carbon dioxide (CO₂) gas, seawater was acidified to pH_NBS 7.6, 7.2 or 6.8. Control treatments were maintained in natural seawater (pH ≈ 8.0). Under normocapnic conditions, burrowing urchins were seen to reduce the sediment uptake of nitrite or nitrate whilst enhancing the release of silicate and phosphate. In acidified (hypercapnic) treatments, the biological control of biogeochemical cycles by urchins was significantly affected, probably through the combined impacts of high CO₂ on nitrifying bacteria, benthic algae and urchin behaviour. This study highlights the importance of considering biological interactions when predicting the consequences of seawater acidification on ecosystem function.     

Enzymatic methods for the determination of pollution in seawater using salt resistant alkaline phosphatase from eggs of the sea urchin Strongylocentrotus intermedius

A new salt resistant alkaline phosphatase from eggs of the sea urchin Strongylocentrotus intermedius (StAP) has been shown to have a unique property to hydrolyze substrate in seawater without loss of enzymatic activity. The enzyme has pH optimum at 8.0–8.5. Model experiments showed various concentrations of copper, zinc, cadmium and lead added to seawater or a standard buffer mixture to inhibit completely the enzyme activity at the concentrations of 15–150 μg/l. StAP sensitivity to the presence in seawater of metals, pesticides, detergents and oil products appears to be considerably less. Samples of seawater taken from aquatic areas of the Troitsy Bay of the Peter the Great Bay, Japan Sea have been shown to inhibit the enzyme activity; the same was shown for the samples of fresh waters. The phosphatase inhibition assay developed proved to be highly sensitive, technically easy-to use allowing to test a great number of samples.