Effects of nitrate and phosphate on growth and C2 toxin productivity of Alexandrium tamarense CI01 in culture

Growth and C2 toxin productivity of a marine dinoflagellate, Alexandrium tamarense CI01 (ATCI01) which predominantly produces C2 toxin, were studied in unialgal batch cultures to optimize the concentrations of nitrate and phosphate for a maximal toxin yield. A range of start concentrations of the two major nutrients was determined in which algal growth was proportional to the nutrient concentrations used. The highest concentrations of nitrate and phosphate in this growth-enhancing range were 264 and 20 μM, respectively. In this concentration range, the C2 toxin yield (μg/l) and cellular toxin content (Qt, fmole per cell) reached a maximum at the lowest end of phosphate (5 μM) and the highest end of nitrate (264 μM). Further increase in the supply of nitrate continued to enhance the toxin yield. Our results indicated that the growth and toxin productivity of this algal strain in batch cultures had distinctly different optimal ranges of nitrate and phosphate concentrations. For a maximum toxin yield, a judicious use of phosphate under a nitrate-replete condition is called for.     

Nitrate and phosphate supplementation to increase toxin production by the marine dinoflagellate Alexandrium tamarense

Alexandrium tamarense toxins have great value in biotechnology research as well as important in connection with shellfish poisoning. The influence of nitrate or nitrate and phosphate supplementation on cell biomass and toxin content were investigated in batch cultures. When cultures at low nitrate (88.2 microM NaNO(3)) were supplemented with 793.8 microM NaNO(3) at day 10 the cell density and cellular toxin contents were increased by 6-29% and 20-76%, respectively, compared with controls, and maximal values were 43,600 cells/ml (day 38) and 0.91 pg/cell (day 31). Supplementation with nitrate at day 14 or with nitrate and phosphate at day 10/14 to the cultures did not increase the cell density compared with the non-supplemented middle nitrate or high phosphate (108 microM NaH(2)PO(4)) cultures, respectively, but increased the cellular toxin contents by an average of 52%. The results showed that supplementation with nitrate or with nitrate and phosphate at different growth phases of the cultures increased toxin yield by an average of 46%. Supplementation with nitrate at selected times to maintain continuous low level of nitrate might contribute to the effective increase of toxin yield of A. tamarense.     

The Impact of the Herbicide Diuron on Photosynthesis in Three Species of Tropical Seagrass

The impact and recovery from exposure to the herbicide diuron [DCMU; 3-(3′,4′-dichlorophenyl)-1,1-dimethylurea] was assessed for three tropical seagrasses, maintained in outdoor aquaria over a 10-day period. Photosynthetic stress was detected using chlorophyll a fluorescence, measured with a Diving-PAM (pulse amplitude modulated fluorometer). Exposure to 10 and 100 μg l⁻¹ diuron resulted in a decline in effective quantum yield (ΔF/F_m^′) within 2 h of herbicide exposure in Cymodocea serrulata, Halophila ovalis and Zostera capricorni. Effective quantum yield also declined over the first 24 h of exposure in H. ovalis at even lower diuron concentrations (0.1 and 1.0 μg l⁻¹). Effective quantum yield in H. ovalis and Z. capricorni was significantly depressed at all diuron concentrations (0.1–100 μg l⁻¹) after 5 days exposure, whereas effective quantum yield in C. serrulata was only significantly lower in plants exposed to highest diuron concentrations (10 and 100 μg l⁻¹). Effective quantum yield depression was present 5 days after plants exposed to 10 and 100 μg l⁻¹ diuron were returned to fresh seawater. These results indicate that exposure to herbicide concentrations present in nearshore Queensland sediments present a potential risk to seagrass functioning.     

The effect of marine colloids on the growth of photosysthetic bacteria

Marine colloids could be an important source of nitrogen for bacteria and photoplankton. But elevated concentration of colloids may stimulate algal growth and lead to red tides in coastal waters. The effects of colloidal organic carbon (COC) concentration on the growth of photosysthetic bacteria (PSB) were investigated under different colloidal treatments in the laboratory. The PSB growth was inversely proportional to COC concentration and was restricted by high-molecular-weight (HMW) colloids (>10 KDa) in treatments with non-nutrient or just inorganic nutrient with low COC concentration (5 μMC). However, the PBS growth was enhanced in the presence of HMW colloids in the treatment with inorganic nutrient and high COC (127 and 255 μMC) or with both inorganic nutrient and low-molecular-weight organic matter. Both bacteria number and bacteria growth ratio increased significantly when the concentration of COC was 5 μMC. Our results suggest that marine colloids can be utilized by bacteria and might regulate primary productivity in coastal waters.     

Methane in the hyporheic zone of a small lowland stream (Sitka, Czech Republic)

Methane concentrations and selected chemical parameters in interstitial water were examined along subsurface flowpath in two subsystems (hyporheic and parafluvial sediments) in the Sitka stream, Czech Republic. Interstitial methane concentrations exhibited a distinct spatial pattern. In the hyporheic downwelling zone where the sediments are relatively well oxygenated due to high hydrologic exchange with the surface water, low interstitial methane concentrations, averaging 9.3 μg CH₄/l, were found. In contrast, upwelling sediments and parafluvial sediments (active channel sediments lateral to the wetted channel) had significantly higher methane concentrations (p < 0.05, and p < 0.01, respectively), averaging 43.2 μg CH₄/l and 160.5 μg CH₄/l, respectively. Dissolved oxygen was the highest where surface water entered hyporheic/parafluvial sediments and decreased with water residence time in the sediments (p < 0.01). Nitrate concentrations decreased along the flowpath and were significantly lower at downstream end of the riffle (p < 0.001). Sulfate concentrations also show a slight decline with the water residence time, but differences were not significant. Effect of both nitrate and sulfate on methanogenesis is also discussed. The interstitial methane concentration significantly increased with surface water temperature (p < 0.001) and was negatively correlated with redox potential (p < 0.01) and dissolved oxygen (p < 0.05).     

PAH contamination of western Irish Sea sediments

Concentrations of the sum of 15 PAHs in 22 surficial sediment samples from the western Irish Sea ranged from below 100 ng g⁻¹ in sandy sediments to a maximum of 1422 ng g⁻¹ in the centre of the mud basin. The concentrations are typical of coastal shelf sediments, but greater than those observed for aquatic sediments remote from known anthropogenic sources. Organic C and the % sediment <15 μm were positively correlated with ΣPAH. ΣPAH was normalised to organic C and particle size (i.e. expressed as ΣPAH/C^org and ΣPAH/%<15 μm) and significant relationships were still observed with organic C and %<15 μm. The results are discussed in the context of using organic C to normalise concentrations of PAHs to assess contamination levels in sediments.     

Changes in phytoplankton following nitrate and phosphate additions to large enclosures in Marion Lake,British Columbia

The addition of nitrate, phosphate and a nitrate phosphate combination to replicated wooden enclosures in Marion Lake resulted in significant increases in primary productivity and algal standing crop in each of the three types of treated enclosures. This indicated that no single nutrient is likely to simultaneously limit the growth of all phytoplankton species present at any one time in the lake epilimnion. The relevance of this to the detergent controversy is discussed. Canadian Contribution to the International Biological Programme No. CCIBP 208.     

Dissolved and particulate trace metals in a Scottish sea loch: an example of a pristine environment?

In the near pristine environment of a silled fjord on the west coast of Scotland samples were taken for the determination of dissolved and particulate trace metals (Fe, Mn, Cu, Ni, Cd, Zn and Pb), together with nutrient and hydrographic data, during 19 surveys carried out over a year. An indication of the pristine nature of the environment are the low concentrations of dissolved silicon, phosphate and nitrate which are considerably lower than those of coastal waters which are subject to larger anthropogenic burdens. Distributions of dissolved Cu, Ni and Cd were found to broadly reflect conservative mixing of freshwater and seawater with both end members having similar concentrations. The concentration of dissolved Cu and Ni in seawater entering upper Loch Linnhe (Cu 0.28 μg l⁻¹; Ni 0.26 μg l⁻¹) was consistent with the 1:1 conservative mix of Irish Sea water and North Atlantic surface water predicted from radio-caesium tracer experiments (Mackay & Baxter, 1985). Atmospheric input of trace metals to upper Loch Linnhe appeared to be a relatively minor term in the mass balance relative to fluvial inputs. Values of distribution coefficients Kd were similar to those previously reported for the coastal environment. Iron showed the strongest affinity for the suspended sediments; with particulate percentages of the total load usually greater than 80%. Lead and Mn showed a similar strong affinity to the particle phase. For Cu, Ni and Zn the mass of the element in the dissolved phase was generally greater than that in the particulate fraction. Cadmium, was least associated with the particles, with typically greater than 90% existing in the dissolved phase.     

Cadmium and manganese distributions in the Hudson River estuary: interannual and seasonal variability

Surface waters collected along the salinity gradient of the Hudson River estuary in four cruises between 1995 and 1997 were size-fractionated into particulate (>0.45 μm), ‘dissolved' (<0.45 μm), colloidal (10 kDa, 0.45 μm) and low molecular weight (<10 kDa) phases. Dissolved Cd concentrations (range: 0.11–1.19 nM) in surface waters of the estuary appear to have decreased fourfold (from an average of 2.36 to 0.61 nM) over a 23-year period, since the initial analysis of samples collected in the 1970s by Klinkhammer and Bender [Estuar. Coastal Shelf Sci. 12 (1981) 629–643]. This interannual decline reflects improvement in sewage treatment and the elimination of industrial Cd sources to the Hudson River estuary. In contrast, dissolved Mn levels (range: 0.033–1.46 μM) have remained relatively constant over the same period of time, suggesting that anthropogenic sources have very limited impact on Mn concentrations in the estuary. The concentrations of both Cd and Mn appeared to strongly depend on the season and/or river discharge. The highest concentrations were detected under low freshwater discharge, implying that limited hydraulic flushing allows a build-up of metals in the water column. Although the decline in Cd levels within the estuary reflects a reduction in the magnitude of anthropogenic inputs, mass balance estimates indicated that current sources of Cd to the estuary include sewage discharges (in the lower estuary around Manhattan) and diagenetic remobilization from industrial Cd deposited in sediments nearly 2 decades ago (in the upper estuary near Foundry Cove). Moreover, under low river discharge, the sources considered in our model (sewage, riverine input, atmospheric deposition, and benthic fluxes) could account for no more than 60% of the Cd exported from the lower estuary to the ocean. This suggests that undefined sources such as ground water and inputs from other watersheds (e.g., Long Island Sound and Newark Bay) may potentially influence the water quality of the New York Harbor. The size-fractionated metal concentrations indicated that most of the traditionally defined ‘dissolved' Cd and Mn consisted of <10 kDa molecular weight species. High molecular weight colloidal species of Mn accounted for about 50% of the dissolved fraction at the riverine end-member and <5% at intermediate and high salinities. Colloidal Cd accounted for <6% of the dissolved phase throughout the estuary. Unlike the non-conservative excess (relative to ideal dilution of river water and seawater) of dissolved Mn observed along the estuary, high molecular weight colloidal Mn appeared to be removed at the head of the estuary. The small contribution of colloidal Cd and Mn to the ‘dissolved' phase suggests that remobilization from suspended particulate phases and/or from sediments occurs through the formation of small molecular weight species.     

Joint FAO/WHO/IOC activities to provide scientific advice on marine biotoxins (research report)

The Joint FAO/WHO/IOC ad hoc Expert Consultation on Biotoxins in Molluscan Bivalves performed risk assessments for a number of biotoxins present in bivalve molluscs. For performing risk assessments, the Expert Consultation categorized the biotoxins into eight distinct groups based on chemical structure. The Expert Consultation established LOAELs for the azaspiracid (AZA), okadaic acid (OA), saxitoxin (STX), and domoic acid (DA) toxin groups. The derived provisional acute RfDs for the AZA, OA, STX, and DA toxin groups were 0.04 μg/kg bw, 0.33 μg/kg bw, 0.7 μg/kg bw, and 100 μg/kg bw, respectively. For the yessotoxin (YTX) group, a NOAEL was established, based on animal studies. Applying a safety factor of 100, a provisional acute RfD of 50 μg/kg bw was suggested for the YTX group. The Expert Consultation considered that the database for cyclic imines, brevetoxins, and pectenotoxins was insufficient to establish provisional acute RfDs for these three toxin groups.     

Pesticide and Herbicide Residues in Sediments and Seagrasses from the Great Barrier Reef World Heritage Area and Queensland Coast

Pesticides and herbicides including organochlorine compounds have had extensive current and past application by Queensland's intensive coastal agriculture industry as well as for a wide range of domestic, public health and agricultural purposes in urban areas. The persistent nature of these types of compounds together with possible continued illegal use of banned organochlorine compounds raises the potential for continued long-term chronic exposure to plants and animals of the Great Barrier Reef. Sediment and seagrass samples were collected from 16 intertidal and 25 subtidal sampling sites between Torres Strait and Townsville, near Mackay and Gladstone, and in Hervey and Moreton Bays in 1997 and 1998 and analysed for pesticide and herbicide residues. Low levels of atrazine (0.1–0.3 μg kg⁻¹), diuron (0.2–10.1 μg kg⁻¹), lindane (0.08–0.19 μg kg⁻¹), dieldrin (0.05–0.37 μg kg⁻¹), DDT (0.05–0.26 μg kg⁻¹), and DDE (0.05–0.26 μg kg⁻¹) were detected in sediments and/or seagrasses. Contaminants were mainly detected in samples collected along the high rainfall, tropical coast between Townsville and Port Douglas and in Moreton Bay. Of the contaminants detected, the herbicide diuron is of most concern as the concentrations detected have some potential to impact local seagrass communities.     

Size effects of suspended particles on gill damage in green-lipped mussel Perna viridis

The green-lipped mussels Perna viridis were exposed to <500 μm suspended solids (SS) with concentrations of 0 (control), 250, 500, 750 and 1000 mg/L for 14 days, followed by transferring to clean, filtered seawater for 28 days. Results of scanning microscopy showed significantly higher damages to the cilia on the frontal surface of the gill filaments than that on the abfrontal surface in both demibranchs. Percent ciliary depletion varied with SS concentrations and time. No sign of recovery of the gill filaments was observed after the mussels were transferred to clean seawater. In a second experiment, mussels were exposed to SS with size range from <63, >125–<250 and >250–<500 μm at 600 mg/L, together with a control (0 mg/L) for 14 days, followed by transferring to clean, filtered seawater for 28 days. Results of scanning microscopy showed significant ciliary damages in both the ascending and descending lamellae under the three particle size groups as compared with the control. Percent depletion of frontal cilia was most serious for the >250–<500 μm size group and least for the <63 μm size group. However, percent depletion of abfrontal cilia was most serious for the >125–<250 μm size group and least for the <63 μm size group. No recovery of ciliary damages was observed. The effects of particle size of suspended sediments on the morphological damages of gill filaments in the green-lipped mussels were discussed.     

Alkaline Phosphatase Activity and Utilization of Dissolved Organic Phosphorus by Algae in Subtropical Coastal Waters

Alkaline phosphatase activity (APA) and the availability of dissolved organic phosphorus (DOP) to marine algae were determined in Xiamen Bay and in algal batch culture systems. Results showed that APA changed with seasons, increasing to the highest value in summer and decreasing to the lowest in autumn and spring in Xiamen Bay. Tests on natural populations of planktonic algae and bacteria community showed that algae were mostly responsible for DOP utilization, while bacteria could not take up DOP compounds. Results from algal batch cultures also supported the above conclusion. Relationships between APA and environmental factors indicated that APA was negatively correlated with phosphorus level such as phosphate and small molecular DOP, and APA played an important role in utilization of DOP by algae. All the results emphasized the ecological significance of DOP in subtropical coastal waters.     

Zinc in Fish, Crabs, Oysters, and Mangrove Flora and Fauna from Cleveland Bay

Zinc in muscle tissue from 28 species of fish, three species of sharks and rays, two species of squid and cuttlefish, and three species of crustacea collected from Cleveland Bay, together with some measurements made on oysters and mangrove flora and fauna are reported. In the fish species the mean Zn concentration in muscle tissue was 4.83 ± 2.82 μg/g wet weight, slightly lower than zinc measurements (7 μg/g wet weight) made in 14 fish species collected from the bay in 1975. Zinc in sharks and rays ranged from 3.5 to 7.2 μg/g wet wt, in squid and cuttlefish 13–16 μg/g wet wt, and in crustacea levels ranged from 14 to 18 μg/g wet wt. Zinc levels in fish varied between species with concentrations well below the ANZECC Maximum Residue Limit of zinc in seafood (150 μg/g wet wt). Although lower zinc concentrations were found in oysters collected from the Townsville Harbour area (2080 μg/g wet wt), compared with previous measurements made 25 years ago at this site, these levels are still above the Maximum Residue Limit for zinc in oysters (1000 μg/g wet wt). Concentrations of zinc in oysters from Orpheus Island (2547 μg/g wet wt), about 74 km away from industrial and urban activity, are also above the safe guideline values. Horseshoe Bay oysters transplanted to Ross Creek accumulated zinc at a rate of about 100 μg/g of oyster tissue per week, suggesting that dissolved zinc levels at this site are elevated, and that oysters rapidly accumulate zinc. Highest concentrations of zinc in mangrove leaves (30–65 μg/g dry wt) occurred in Osbornia octodonta, Exocaria agalocha, and Aegialitus annulata, compared with Ceriops tagal, and Avicennia marina (5–10 μg/g dry wt). No significant difference in zinc concentration occurred between leaves and litter fall for most of these species, with the sole exception of Exocaria, which showed almost a twofold increase in concentration. In seven species of mangrove fauna levels of zinc were very uniform and close to 50 μg/g (dry wt) Telescopium telescopium from the mouth of the Ross Estuary was the exception with levels at 400 μg/g (dry wt).     

Nitrate and herbicide loading in two groundwater basins of Illinois'' sinkhole plain

This investigation was designed to estimate the mass loading of nitrate (NO₃⁻) and herbicides in spring water discharging from groundwater basins in an agriculturally dominated, mantled karst terrain. The loading was normalized to land use and NO₃⁻ and herbicide losses were compared to estimated losses in other agricultural areas of the Midwestern USA. Our study area consisted of two large karst springs that drain two adjoining groundwater basins (total area of 37.7 km²) in southwestern Illinois' sinkhole plain, USA. The springs and stream that they form were monitored for almost 2 years. Nitrate–nitrogen (NO₃–N) concentrations at three monitoring sites were almost always above the background concentration (1.9 mg/l). NO₃–N concentrations at the two springs ranged from 1.08 to 6.08 with a median concentration of 3.61 mg/l. Atrazine and alachlor concentrations ranged from <0.01 to 34 μg/l and <0.01 to 0.98 μg/l, respectively, with median concentrations of 0.48 and 0.12 μg/l, respectively. Approximately 100,000 kg/yr of NO₃–N, 39 kg/yr of atrazine, and 2.8 kg/yr of alachlor were discharged from the two springs. Slightly more than half of the discharged NO₃⁻ came from background sources and most of the remainder probably came from fertilizer. This represents a 21–31% loss of fertilizer N from the groundwater basins. The pesticide losses were 3.8–5.8% of the applied atrazine, and 0.05–0.08% of the applied alachlor. The loss of atrazine adsorbed to the suspended solid fraction was about 2 kg/yr, only about 5% of the total mass of atrazine discharged from the springs.     

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.     

On the biophilic nature of iodine in seawater

Vertical profiles of concentrations of iodate- and total-iodine have been measured at thirty stations in the Pacific, Atlantic and Antarctic Oceans. The salinity-normalised iodine profiles are indicative of both iodine removal and iodate reduction in the euphotic zone. Thus, surface waters appear to be depleted in iodate-iodine (by 0.03?0.22 μM) but less so in total-iodine (by<0.01?0.06 μM) when compared with the near-constant iodine concentrations (～0.46 μM) at depth. Graphs of specific total-iodine versus specific phosphate fit a linear model well and lie within a narrow envelope for all stations, suggesting a direct coupling of iodine and nutrients during assimilation/regeneration. The I/C atom ratio calculated from these hydrographic data (1.0 × 10^?4) agrees well with contemporary plankton compositions (I/C= 1.4 (±0.8) × 10^?4). Similar graphs involving specific iodate also fit a linear model well. However, their gradients vary from station to station leading to a variability in I/C interconversion ratio, analogous to the variability of Redfield nutrient ratios for coastal waters. This variation is attributed to changes in both productivity and nitrate availability. Pacific deep waters contain anomalously high total-iodine concentrations which may reflect regional differences of I/P ratio in surface waters or else diffusion of iodine from bottom sediments.     

Characterization and Application of Dried Plants to Remove Heavy Metals,Nitrate, and Phosphate Ions from Industrial Wastewaters

Low cost adsorbents were prepared from dried plants for the removal of heavy metals, nitrate, and phosphate ions from industrial wastewaters. The efficiency of these adsorbents was investigated using batch adsorption technique at room temperature. The dried plant particles were characterized by N₂ at 77 K adsorption, scanning electron microscopy, energy dispersive X‐ray spectroscopy, Fourier transform infrared spectroscopy, and phytochemical screening. The adsorption experiments showed that the microparticles of the dried plants presented a good adsorption of heavy metals, phosphate, and nitrate ions from real wastewaters. This adsorption increased with increasing contact time. The equilibrium time was found to be 30 min for heavy metals and nitrate ions and 240 min for phosphate ions. After the adsorption process, the Pb(II) concentrations, as well as those of Cd(II), Cu(II), and Zn(II) were below the European drinking water norms concentrations. The percentage removal of heavy metals, nitrates, and phosphates from industrial wastewaters by dried plants was ～94% for Cd²⁺, ～92% for Cu²⁺, ～99% for Pb²⁺, ～97% for Zn²⁺, ～100% for ${\rm NO}_{{\rm 3}}^{{-} } $ and ～77% for ${\rm PO}_{{\rm 4}}^{3{-} } $ ions. It is proved that dried plants can be one alternative source for low cost absorbents to remove heavy metals, nitrate, and phosphate ions from municipal and industrial wastewaters.     

Levels of chlorinated hydrocarbons and trace metals in bivalves and nearshore sediments from the Dominican Republic

Bivalves (Crassostrea rizhophorae, Isognomon alatus, Codakia orbicularis Linné, and Tellina fausta) and sediment samples collected from 12 locations along the Dominican Republic coastline were used to assess the degree of nearshore contamination by selected chlorinated hydrocarbons and heavy metals. Concentrations of total chlordanes, DDTs and PCBs ranged from 0.51 to 7.47 ng/g, < MDL to 30.9 ng/g and 11.3 to 82.3 ng/g, and from < MDL to 1.73 ng/g, 0.21 to 12.5 ng/g and 0.46 to 41.9 ng/g in bivalves and surface sediments, respectively. The concentration ranges of predominantly anthropogenic Cu, Pb and Zn were 3.08–866 μg/g, 0.08–1.46 μg/g and 22.9–4380 μg/g in bivalve soft parts and 1.01–111 mg/g, 0.420–134 μg/g and 1.96–277 μg/g in surface sediments, respectively. Mercury concentrations ranged from 0.290 to 7.02 μg/g and from 0.096 to 0.565 μg/g in both matrices. The relatively high Hg concentrations found in bivalves from two stations in Bahia Samana, near to a known source of past Hg contamination in the sediments, require a more in-depth survey of the area. In general the concentrations of selected chlorinated hydrocarbons and trace metals, compared with those reported for the same matrices in the Caribbean and west Atlantic coastal areas, show at present the existence of only very localized contamination problems in the Dominican Republic.