Nutrient couplings between on-site sewage disposal systems, groundwaters, and nearshore surface waters of the Florida Keys

We performed a one-year study to determine the effects of on-site sewage disposal systems (OSDS, septic tanks) on the nutrient relations of limestone groundwaters and nearshore surface waters of the Florida Keys. Monitor wells were installed on canal residences with OSDS and a control site in the Key Deer National Wildlife Refuge on Big Pine Key. Groundwater and surface water samples were collected monthly during 1987 and analyzed for concentrations of dissolved inorganic nitrogen (DIN = NO_f3/sup- + NO_f2/sup- + NH_4/su+), soluble reactive phosphate (SRP), temperature and salinity. Significant nutrient enrichment (up to 5000-fold) occurred in groundwaters contiguous to OSDS; DIN was enriched an average of 400-fold and SRP some 70-fold compared to control groundwaters. Ammonium was the dominant nitrogenous species and its concentration ranged from a low of 0.77 μM in control wells to 2.75 mM in OSDS-enriched groundwaters. Concentrations of nitrate plus nitrite were also highly enriched and ranged from 0.05 μM in control wells to 2.89 mM in enriched groundwaters. Relative to DIN, concentrations of SRP were low and ranged from 30 nM in control wells to 107 μM in enriched groundwaters. N : P ratios of enriched groundwaters were consistently > 100 and increased with increasing distance from the OSDS, suggesting significant, but incomplete, adsorption of SRP by subsurface flow through carbonate substrata. Nutrient concentrations of groundwaters also varied seasonally and were approximately two-fold higher during the winter (DIN = 1035 μM; SRP = 10.3 μM) compared to summer (DIN = 470 μM; SRP = 4.0 μM). In contrast, surface water nutrient concentrations were two-fold higher during the summer (DIN = 5.0 μM; SRP = 0.50 μM) compared to winter (DIN = 2.5 μM; SRP = 0.15 μM). Direct measurement of subsurface groundwater flow rate indicated that tides and increased groundwater recharge enhanced flow some two-fold and six-fold, respectively. Accordingly, the observed seasonal coupling of OSDS-derived nutrients from groundwaters to surface waters is maximum during summer because of seasonally maximum tides and increased hydraulic head during the summer wet season. The yearly average benthic flux of anthropogenic DIN into contiguous canal surface waters is 55 mmol m^-2 day^-1, a value some five-fold greater than the highest rate of benthic N-fixation measured in carbonate-rich tropical marine waters.     

Phytoplankton nutrient deficiency in lakes of the McMurdo dry valleys, Antarctica

1. The influence of inorganic nitrogen and phosphorus enrichment on phytoplankton photosynthesis was investigated in Lakes Bonney (east and west lobes), Hoare, Fryxell and Vanda, which lie in the ablation valleys adjacent to McMurdo Sound, Antarctica. Bioassay experiments were conducted during the austral summer on phytoplankton populations just beneath the permanent ice cover in all lakes and on populations forming deep-chlorophyll maxima in the east and west lobes of Lake Bonney. 2. Phytoplankton photosynthesis in surface and mid-depth (13 m) samples from both lobes of Lake Bonney were stimulated significantly (P < 0.01) by phosphorus enrichment (2 μM) with further stimulation by simultaneous phosphorus plus NH₄⁺ (20 μM) enrichment. Similar trends were observed in deeper waters (18 m) from the east lobe of Lake Bonney, although they were not statistically significant at P < 0.05. Photosynthesis in this lake was never enhanced by the addition of 20 μM NH₄⁺ alone. Simultaneous addition of phosphorus plus nitrogen stimulated photosynthesis significantly (P < 0.01) in both Lake Hoare and Lake Fryxell. No nutrient response occurred in Lake Vanda, where activity in nutrient-enriched samples was below unamended controls; results from Lake Vanda are suspect owing to excessively long sample storage in the field resulting from logistic constraints. 3. Ambient dissolved inorganic nitrogen (DIN) (NH4⁺+ NO₂^?+ NO₃^?): soluble reactive phosphorus (SRP) ratios partially support results from bioassay experiments indicating strong phosphorus deficiency in Lake Bonney and nitrogen deficiency in Lakes Hoare and Fryxell. DIN : SRP ratios also imply phosphorus deficiency in Lake Vanda, although not as strong as in Lake Bonney. Particulate carbon (PC): particulate nitrogen (PN) ratios all exceed published ratios for balanced phytoplankton growth, indicative of nitrogen deficiency. 4. Vertical nutrient profiles in concert with low advective flux, indicate that new (sensu Dugdale & Goering, 1967) phytoplankton production in these lakes is supported by upward diffusion of nutrients from deep nutrient pools. This contention was tested by computing upward DIN : SRP flux ratios across horizontal planes located immediately beneath each chlorophyll maximum and about 2 m beneath the ice (to examine flux to the phytoplankton immediately below the ice cover). These flux ratios further corroborated nutrient bioassay results and bulk DIN : SRP ratios indicating phosphorus deficiency in Lakes Bonney and Vanda and potential nitrogen deficiency in Lakes Hoare and Fryxell. 5. Neither biochemical reactions nor physical processes appear to be responsible for differences in nutrient deficiency among the study lakes. The differences may instead be related to conditions which existed before or during the evolution of the lakes.     

Macroalgal blooms on southeast Florida coral reefs: I. Nutrient stoichiometry of the invasive green alga Codium isthmocladum in the wider Caribbean indicates nutrient enrichment

Invasive blooms of the siphonaceous green algae Codium spp. have been considered a symptom of coastal eutrophication but, to date, only limited biochemical evidence supports a linkage to land-based nutrient pollution. Beginning in the summer of 1990, spectacular blooms of unattached Codium isthmocladum developed on deep coral reef habitats in southern Palm Beach County and northern Broward County, and in subsequent years, attached populations formed on reefs in northern Palm Beach County.To better understand the nutrition of these HABs, we collected C. isthmocladum and other reef macroalgae from various locations in southeast Florida as well as the wider Caribbean region for tissue C:N:P analysis in order to gauge variability in the type and degree of N- and/or P-limited growth. Widespread nutrient enrichment in floridian C. isthmocladum populations was evidenced by significantly higher tissue P (0.06% versus 0.04% of dry weight) and lower C:N (12 versus 19), C:P (425 versus 980), and N:P (35 versus 50) ratios compared to more nutrient-depleted Caribbean populations. To determine nutrient availability on southeast Florida's reefs, we sampled near-bottom waters at a variety of locations for DIN (NH₄⁺ + NO₃⁻ + NO₂⁻) and SRP analysis. In general, concentrations of NH₄⁺, NO₃⁻and SRP were all high on southeast Florida's reefs compared to values reported for Caribbean coral reefs. Although summertime upwelling provides episodic NO₃⁻ and SRP enrichment to reefs in southeast Florida, these transient nutrient pulses have not historically supported C. isthmocladum blooms.We suggest that the widespread P enrichment of C. isthmocladum tissue and water column DIN:SRP ratios <16:1 in southeast Florida drive this system toward N limitation where low level NH₄⁺ enrichment becomes of paramount importance. Hence, the recent C. isthmocladum blooms appear to be supported by increasing land-based nutrient pollution, particularly, sewage that is enriched in NH₄⁺ and SRP at a low N:P ratio (<10:1) critical to sustaining balanced growth during bloom formation.     

Ulva rigida (Ulvales,Chlorophyta) tank culture as biofilters for dissolved inorganic nitrogen from fishpond effluents

Ulva rigida was cultivated in 7501 tanks at different densities with direct and continuous inflow (at 2, 4, 8 and 12 volumes d^–1) of the effluents from a commercial marine fishpond (40 metric tonnes, Tm, of Sparus aurata, water exchange rate of 16 m³ Tm^–1) in order to assess the maximum and optimum dissolved inorganic nitrogen (DIN) uptake rate and the annual stability of the Ulva tank biofiltering system. Maximum yields (40 g DW m^–2 d^–1) were obtained at a density of 2.5 g FW 1^–1 and at a DIN inflow rate of 1.7 g DIN m^–2 d^–1. Maximum DIN uptake rates were obtained during summer (2.2 g DIN M^–2 d^–1), and minimum in winter (1.1 g DIN m^–2 d^–1) with a yearly average DIN uptake rate of 1.77 g DIN m^–2 d^–1 At yearly average DIN removal efficiency (2.0 g DIN m^–2 d^–1, if winter period is excluded), 153 m² of Ulva tank surface would be needed to recover 100% of the DIN produced by 1 Tm of fish.Abbreviations DIN= dissolved inorganic nitrogen (NH _inf4 ^sup+ + NO _inf3 ^sup– + NO _inf2 ^sup– ); - FW= fresh weight; - DW= dry weight; - PFD= photon flux density; - V= DIN uptake rate     

Determining trophic state in Lake Whatcom,Washington (USA), a soft water lake exhibiting seasonal nitrogen limitation

We evaluated an eleven year data set to assess trophic state and nutrient limitation in Lake Whatcom, an oligotrophic, soft water, chain lake located in the Puget Sound lowlands of Washington (U.S.A.). Although total phosphorus (TP) and soluble reactive phosphate (SRP) concentrations were relatively low throughout the lake, there were significant differences between the northern basin (Site 1) and the other sampling sites (Sites 2–4). Nonparametric correlation coefficients (Kendall's ) were highest between chlorophyll (CHL), Secchi depth (SD), total nitrogen (TN), and dissolved inorganic nitrogen (DIN). Late summer algal biomass correlated best with DIN and TP. Trophic State Indices based on TP, TN, CHL and SD revealed that although algal growth was most likely phosphorus limited throughout the year, the northern basin of the lake may have developed nitrogen co-limitation during late summer and fall. During this period, N/P ratios were often less than 20, and in 1998 the epilimnetic DIN concentrations dropped below 20 g l^–1 while DIN/TP ratios fell below 4. Reviews of the literature suggest that while co-limitation by phosphorus and nitrogen is fairly common in unproductive lakes, the patterns seen in Lake Whatcom were more similar to those reported for eutrophic lakes experiencing secondary nitrogen limitation resulting from excess phosphorus loading.     

Variability in the water chemistry of shallow ponds in southeast England, with special reference to the seasonality of nutrients and implications for modelling trophic status

The variability in water chemistry of samples taken on a monthly basis (October 1990–December 1991) from 31 shallow, artificial ponds in southeast England was examined. The survey revealed great within-year variation in the concentrations of nutrients. Total phosphorus (TP) concentrations displayed no overall marked seasonal pattern, although many sites experienced summer peaks with a simultaneous increase in soluble reactive phosphorus (SRP) concentrations, indicating that sediment P release occurred. SRP and nitrate (NO₃–N) concentrations displayed a marked seasonal pattern similar to that observed in deep, stratifying lakes, with 55% and 94% of the sites surveyed, respectively, experiencing a decline in concentrations in the spring, maintaining low levels throughout summer and the highest levels occurring in winter. Dissolved silica (SiO₂–(Si) also displayed a marked seasonal distribution with a spring decline in concentrations associated with diatom growth, followed by an increase in the summer.The study demonstrated that intra-annual variability in nutrient concentrations is high and tends to be greatest in the most enriched waters. Thus, a high sampling frequency is required to provide representative annual mean data. Furthermore, annual means rather than winter–spring means provide more appropriate estimates of TP and SRP in these waters, owing to the importance of internal cycling of nutrients in summer. The findings are especially relevant to sampling strategy design and the averaging of seasonal water chemistry data for use in predictive models of lake trophic status.     

Septic systems contribute to nutrient pollution and harmful algal blooms in the St. Lucie Estuary,Southeast Florida,USA

Nutrient enrichment is a significant global-scale driver of change in coastal waters, contributing to an array of problems in coastal ecosystems. The St. Lucie Estuary (SLE) in southeast Florida has received national attention as a result of its poor water quality (elevated nutrient concentrations and fecal bacteria counts), recurring toxic Microcystis aeruginosa blooms, and its proximity to the northern boundary of tropical coral species in the United States. The SLE has an artificially large watershed comprised of a network of drainage canals, one of which (C-44) is used to lower the water level in Lake Okeechobee. Public attention has primarily been directed at nutrient inputs originating from the lake, but recent concern over the importance of local watershed impacts prompted a one-year watershed study designed to investigate the interactions between on-site sewage treatment and disposal systems (OSTDS or septic systems), groundwaters, and surface waters in the SLE and nearshore reefs. Results provided multiple lines of evidence of OSTDS contamination of the SLE and its watershed: 1) dissolved nutrients in groundwaters and surface waters were most concentrated adjacent to two older (pre-1978) residential communities and the primary canals, and 2) sucralose was present in groundwater at residential sites (up to 32.0 μg/L) and adjacent surface waters (up to 5.5 μg/L), and 3) δ¹⁵N values in surface water (+7.5 ^o/_oo), macroalgae (+4.4 ^o/_oo) and phytoplankton (+5.0 ^o/_oo) were within the published range (>+3 ^o/_oo) for sewage N and similar to values in OSTDS-contaminated groundwaters. Measured δ¹⁵N values in M. aeruginosa became increasingly enriched during transport from the C-44 canal (∼5.8 ^o/_oo) into the mid-estuary (∼8.0 ^o/_oo), indicating uptake and growth on sewage N sources within the urbanized estuary. Consequently, there is a need to reduce N and P loading, as well as fecal loading, from the SLE watershed via septic-to-sewer conversion projects and to minimize the frequency and intensity of the releases from Lake Okeechobee to the SLE via additional water storage north of the lake. These enhancements would improve water quality in both the SLE and Lake Okeechobee, reduce the occurrence of toxic harmful algal blooms in the linked systems, and improve overall ecosystem health in the SLE and downstream reefs.     

In vitro differentiation in callus cultures of moth bean,Vigna aconitifolia (JACQ) marechal

Callus cultures of two cultivars of Vigna aconitifolia (IPCMO-926, RDM-120) were raised and their growth and differentiation studied. In IPCMO-926 callus cultures, numerous shoot buds differentiated on MS medium with BA (0.4–22.2 μM) alone or in combination with IAA (5.7 μM). In RDM-120 best differentiation of shoot buds was observed on a medium with K (23.2 μM) and IAA (5.7 μM). Kinetin alone, however, induced rhizogenesis in callus cultures. In suspension cultures of IPCMO-926 embryoids differentiated on MS medium with K (0.5 μM) and 2,4-D (0.4 and 0.9 μM).     

Studies on a gram-positive hydrogen bacterium,Nocardia opaca 1 b

Nocardia opaca strain 1 b has a NAD-dependent hydrogenase (hydrogen dehydrogenase). The enzyme has been purified from autotrophically grown cells and tested for optimal assay conditions and stability. The purification procedure involved protamine sulfate treatment, ammonium sulfate precipitation, and separation by DEAE-cellulose and Sephadex G-200 chromatography and resulted in a 63-fold increase of specific activity at a 11.7% enzyme recovery. The final specific activity was 103 μmoles H₂/min·mg protein. The purified enzyme was dependent on nickel and magnesium ions at 0.5 and 5.0 mM concentrations, respectively, as well as flavin mononucleotide at a 5–10 μM concentration. Straight enzyme kinetics were achieved by preincubating the enzyme in the presence of NADH₂. A high stability of the enzyme was observed in 0.1 M potassium phosphate buffer, pH 6.5, in the presence of 0.5 mM nickel and 5 mM magnesium ions under hydrogen atmosphere. Even under air the enzyme was remarkably stable, although less than under hydrogen. From double reciprocal plots of substrate saturation curves the Michaelis-Menten constants were calculated: For saturating NAD-concentration the K _m was 0.063 mM H₂ and for saturating hydrogen concentration the K _m was 0.123 mM NAD.     

Expression of the NAD-dependent FDH1 β-subunit from Methylobacterium extorquens AM1 in Escherichia coli and its characterization

The efficient regeneration of nicotinamide cofactors is an important process for industrial applications because of their high cost and stoichiometric requirements. In this study, the FDH1 β-subunit of NAD-dependent formate dehydrogenase from Methylobacterium extorquens AM1 was heterologously expressed in Escherichia coli. It showed water-forming NADH oxidase (NOX-2) activity in the absence of its α-subunit. The β-subunit oxidized NADH and generated NAD⁺. The enzyme showed a low NADH oxidation activity (0.28 U/mg enzyme). To accelerate electron transfer from the enzyme to oxygen, four electron mediators were tested; flavin mononucleotide, flavin adenine dinucleotide, benzyl viologen (BV), and methyl viologen. All tested electron mediators increased enzyme activity; addition of 250 μM BV resulted in the largest increase in enzyme activity (9.98 U/mg enzyme; a 35.6-fold increase compared with that in the absence of an electron mediator). Without the aid of an electron mediator, the enzyme had a substrate-binding affinity for NADH (K _m) of 5.87 μM, a turnover rate (k _cat) of 0.24/sec, and a catalytic efficiency (k _cat/K _m) of 41.31/mM/sec. The addition of 50 μM BV resulted in a 22.75-fold higher turnover rate (k _cat, 5.46/sec) and a 2.64-fold higher catalytic efficiency (k _cat/K _m, 107.75/mM/sec).     

Studies on choline acetyltransferase isolated from human brain

The purified choline acetyltransferase from human striatal tissue was found to have aK _m value of 8 μM for acetyl-coenzyme A and 250 μM for choline. The predominant enzyme component has a molecular weight of about 67,000 daltons, measured by molecular filtration through Sephadex G-100. In a sucrose-density gradient, the enzyme cosedimented with bovine serum albumin with an estimatedS-value of 4.5. The enzyme activity was enhanced 2- to 3-fold by KCl, NaCl, (NH₄)₂SO₄, and chelating agents like EDTA or EGTA. Cupric sulfate (0.1 mM) inhibited the enzyme activity almost completely. This inhibition was circumvented by increasing concentrations of enzyme protein, dithiothreitol, and EDTA, but not by the substrates, histidine, or imidazole.     

PHYSIOLOGICAL INDICATORS OF NUTRIENT DEFICIENCY IN CLADOPHORA (CHLOROPHYTA) IN THE CLARK FORK OF THE COLUMBIA RIVER,MONTANA1

Cellular nutrient concentrations and nutrient uptake rates of Cladophora glomerata (L.) Kuetzing were determined during summer and fall in 1989–1990 at a site on the upper Clark Fork of the Columbia River, Montana. Both physiological tests indicated that Cladophora growth is likely to be limited by nitrogen during late summer-early fall. Maximum uptake rates of ammonia-N and nitrate-N were 5935–6991 and 507–984 μg · g DW^?1· h^?1, respectively, during July–October when dissolved inorganic nitrogen (DIN) concentrations in the river were less than 10 μg · L^?1. During November-December, when DIN was 72–376 μg · L^?1, maximum ammonia-N uptake was 1137–1633 μg · g DW^?1· h^?1 and maximum nitrate-N uptake was 0–196 μg · g DW^?1· h^?1. Cellular nitrogen during summer–early fall was 0.78–1.80% of Cladophora dry weight, frequently at or below 1.1%, a level suggested as a critical minimum N concentration for maximum growth. In contrast, cellular P was 0.18–0.36% of dry weight, 3–6 times the suggested critical P concentration of 0.06%. Molar ratios of cellular N:P (< 16:1) and DIN: SRP (< 4:1) during late summer-early fall also indicated potential N limitation. Cellular N and P from Cladophora collected from a second site influenced by a municipal wastewater discharge in 1990 displayed similar seasonal trends. At both sites, seasonal fluctuations in DIN were closely tracked by changes in cellular N, Cellular P, however, increased through the growing season despite declining levels of SRP in the river.     

Regional analysis of inorganic nitrogen yield and retention in high-elevation ecosystems of the Sierra Nevada and Rocky Mountains

Yields and retention of dissolved inorganic nitrogen (DIN: NO₃ ^– + NH₄ ⁺) and nitrate concentrations in surface runoff are summarized for 28 high elevation watersheds in the Sierra Nevada of California and Rocky Mountains of Wyoming and Colorado. Catchments ranged in elevation from 2475 to 3603 m and from 15 to 1908 ha in area. Soil cover varied from 5% to nearly 97% of total catchment area. Runoff from these snow-dominated catchments ranged from 315 to 1265 mm per year. In the Sierra Nevada, annual volume-weighted mean (AVWM) nitrate concentrations ranged from 0.5 to 13 M (overall average 5.4 M), and peak concentrations measured during snowmelt ranged from 1.0 to 38 M. Nitrate levels in the Rocky Mountain watersheds were about twice those in the Sierra Nevada; average AVWM NO₃ ^– was 9.4 M and snowmelt peaks ranged from 15 to 50 M. Mean DIN loading to Rocky Mountain watersheds, 3.6 kg ha^–1 yr^–1, was double the average measured for Sierra Nevada watersheds, 1.8 kg ha^–1 yr^–1. DIN yield in the Sierra Nevada, 0.69 kg ha^–1 yr^–1, was about 60% that measured in the Rocky Mountains, 1.1 kg ha^–1 yr^–1. Net inorganic N retention in Sierra Nevada catchments was 1.2 kg ha^–1 yr^–1 and represented about 55% of annual DIN loading. DIN retention in the Rocky Mountain catchments was greater in absolute terms, 2.5 kg ha^–1 yr^–1, and as a percentage of DIN loading, 72%.A correlation analysis using DIN yield, DIN retention and surface water nitrate concentrations as dependent variables and eight environmental features (catchment elevation, slope, aspect, roughness, area, runoff, soil cover and DIN loading) as independent variables was conducted. For the Sierra Nevada, elevation and soil cover had significant (p > 0.1) Pearson product moment correlations with catchment DIN yield, AVWM and peak snowmelt nitrate concentrations and DIN retention rates. Log-linear regression models using soil cover as the independent variable explained 82% of the variation in catchment DIN retention, 92% of the variability in AVWM nitrate and 85% of snowmelt peak NO₃ ^–. In the Rocky Mountains, soil cover was significantly (p < 0.05) correlated with DIN yield, AVWM NO₃ ^– and DIN retention expressed as a percentage of DIN loading (%DIN retention). Catchment mean slope and terrain roughness were positively correlated with steam nitrate concentrations and negatively related to %DIN retention. About 91% of the variation in DIN yield and 79% of the variability in AVWM NO₃ ^– were explained by log-linear models based on soil cover. A log-linear regression based on soil cover explained 90% of the variation of %DIN retention in the Rocky Mountains.     

Ammonium and nitrate as nitrogen sources in two Eriophorum species

Summary We compared ammonium and nitrate nutrition in Eriophorum scheuchzeri and E. vaginatum, two Alaskan sedges that are native to high- and low-fertility sites, respectively. When grown in solution culture, the two species were similar in their kinetics of NH _inf4 ^sup+ NO _inf3 ^sup- absorption: at nitrogen concentrations below 50 M, net NH _inf4 ^sup+ and NO _inf3 ^sup- were absorbed at similar rates, but at higher concentrations, net uptake of NO _inf3 ^sup- was significantly faster than that of NH _inf4 ^sup+ . The two species also showed similar abilities to assimilate NO _inf3 ^sup- . Growth of E. vaginatum under NO _inf3 ^sup- nutrition was only slightly less than that under NH _inf4 ^sup+ . The observed similarities between these species from contrasting edaphic habitats indicate that factors other than tissue-specific rates of nitrogen acquisition and assimilation may underlie local adaptation to soil N fertility. Moreover, the capacity of these species to exploit NO _inf3 ^sup- as a N source supports the view that NO _inf3 ^sup- availability may be significant even in wet, acidic, arctic soils.     

Predominance of phytoplankton-derived dissolved and particulate organic carbon in a highly eutrophic tropical coastal embayment (Guanabara Bay,Rio de Janeiro,Brazil)

We investigate the carbon dynamics in Guanabara Bay, an eutrophic tropical coastal embayment surrounded by the megacity of Rio de Janeiro (southeast coast of Brazil). Nine sampling campaigns were conducted for dissolved, particulate and total organic carbon (DOC, POC and TOC), dissolved inorganic carbon (DIC), partial pressure of CO₂ (pCO₂), chlorophyll a (Chl a), pheo-pigments and ancillary parameters. Highest DOC, POC and Chl a concentrations were found in confined-shallow regions of the bay during the summer period with strong pCO₂ undersaturation, and DOC reached 82 mg L^?1, POC 152 mg L^?1, and Chl a 800 μg L^?1. Spatially and temporally, POC and DOC concentrations varied positively with total pigments, and negatively with DIC. Strong linear correlations between these parameters indicate that the production of TOC translates to an equivalent uptake in DIC, with 85% of the POC and about 50% of the DOC being of phytoplanktonic origin. Despite the shallow depths of the bay, surface waters were enriched in POC and DOC relative to bottom waters in periods of high thermohaline stratification. The seasonal accumulation of phytoplankton-derived TOC in the surface waters reached about 105 g C m^?2 year^?1, representing between 8 and 40% of the net primary production. The calculated turnover time of organic carbon was 117 and 34 days during winter and summer, respectively. Our results indicate that eutrophication of coastal bays in the tropics can generate large stocks of planktonic biomass and detrital organic carbon which are permanently being produced and partially degraded and buried in sediments.     

Anthropogenic nutrient enrichment of seagrass and coral reef communities in the Lower Florida Keys: discrimination of local versus regional nitrogen sources

Land-based nutrient pollution represents a significant human threat to coral reefs globally. We examined this phenomenon in shallow seagrass and coral reef communities between the Content Keys (southern Florida Bay) and Looe Key (south of Big Pine Key) in the Lower Florida Keys by quantifying the role of physical forcing (rainfall, wind, tides) and water management on mainland South Florida to nutrient enrichment and blooms of phytoplankton, macroalgae, and seagrass epiphytes. Initial studies (Phase I) in 1996 involved daily water quality sampling (prior to, during, and following physical forcing events) at three stations (AJ, an inshore area directly impacted by sewage discharges; PR, a nearshore patch reef located inshore of Hawk Channel; and LK, an offshore bank reef at Looe Key) to assess the spatial and temporal patterns in advection of land-based nutrients to the offshore reefs. Concentrations of dissolved inorganic nitrogen (DIN=NH₄⁺+NO₃⁻+NO₂⁻), soluble reactive phosphorus (SRP), and chlorophyll a increased at PR and LK following a wind event (∼15 knots, northeast) in mid-February. The highest DIN (mostly NH₄⁺) and SRP concentrations of the entire study occurred at the inshore AJ during an extreme low tide in March. Following the onset of the wet season in May, mean NH₄⁺ and chlorophyll a concentrations increased significantly to maximum seasonal values at PR and LK during summer; relatively low concentrations of NO₃⁻ and a low f-ratio (NO₃⁻/NH₄⁺+NO₃⁻) at all stations during summer do not support the hypothesis that the seasonal phytoplankton blooms resulted from upwelling of NO₃⁻. A bloom of the seagrass epiphyte Cladosiphon occidentalis (phaeophyta) followed the onset of the rainy season and increased NH₄⁺ concentrations at LK, resulting in very high epiphyte:blade ratios (∼3:1) on Thalassia testudinum. Biomass of macroalgae increased at all three stations from relatively low values (<50 g dry wt m⁻²) in winter and early spring to higher values (∼100-300 g dry wt m⁻²) typical of eutrophic seagrass meadows and coral reefs following the onset of the rainy season. The mean δ¹⁵N value of Laurencia intricata (rhodophyta) during 1996 at AJ (+4.7‰) was within the range reported for macroalgae growing on sewage nitrogen; lower values at the more offshore PR (+3.1‰) and LK (+2.9‰) were at the low end of the sewage range, indicating an offshore dilution of the sewage signal during the 1996 study. However, transient increases in δ¹⁵N of Cladophora catanata (chlorophtyta) from ~+2% to +5% at LK concurrent with elevated NH₄⁺ concentrations following rain and/or wind events in May and July suggest episodic advection of sewage nitrogen to the offshore LK station. The Phase II study involved sampling of macroalgae for δ¹⁵N along a gradient from the Content Keys through Big Pine Key and offshore to LK in the summer wet season of 2000 and again in the drought of spring 2001. During the July 2000 sampling, macroalgae in nearshore waters around Big Pine Key had elevated δ¹⁵N values (~+4‰) characteristic of sewage enrichment; lower values (~+2‰) at LK were similar to values reported for macroalgae in upstream waters of western Florida Bay influenced by nitrogen-rich Everglades runoff. That pattern contrasted with the drought sampling in March 2001, when δ¹⁵N values of macroalgae were elevated (+6‰) to levels characteristic of sewage enrichment over a broad spatial scale from the Content Keys to LK. These results suggest that regional-scale agricultural runoff from the mainland Everglades watersheds as well as local sewage discharges from the Florida Keys are both significant nitrogen sources supporting eutrophication and algal blooms in seagrass and coral reef communities in the Lower Florida Keys. Hydrological and physical forcing mechanisms, including rainfall, water management on the South Florida mainland, wind, and tides, regulate the relative importance and variability of these anthropogenic nitrogen inputs over gradients extending to the offshore waters of the Florida Reef Tract.     

Antagonistic effects of Satureja hortensis essential oil against AFB,on human lymphocytes in vitro

Satureja hortensis L. (Lamiaceae) has been used as a folk remedy to treat various such as cramps, muscle pains, nausea, indigestion, diarrhea, and infectious diseases. In this study, the antagonistic effects of essential oil of S. hortensis (SHE) were studied against aflatoxin B₁ (AFB₁) in human lymphocytes in vitro. The analysis of the essential oil was performed by using Gas chromatography-mass spectrometry (GC-MS). Anti-genotoxic effects of the SHEs was evaluated using sister chromatid exchange (SCE), micronuclei (MN) tests against AFB1. Also level of malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione peroxidase (GPx) activities used to determine the anti-oxidative effects of the SHEs. This result showed AFB₁ (5 μM) increased the frequencies of SCE, MN and the level of MDA. AFB1 at the same concentration decreased the activities of SOD and GPx. However, different concentrations of SHE with AFB¹ decreased the frequency of SCE and MN and level of MDA and also increased the activities of SOD and GPx significantly. Especially, the 1.0, 1.5, 2.0 μL dose of SHE are more effective than other doses. The results of this experiment have clearly shown that SHE has strong antioxidative and antigenotoxic effects, these biological activities of SHEs can be due to its component.     

15N evidence for the origin and cycling of inorganic nitrogen in a small Amazonian catchment

The ¹⁵N composition of the dominant form of dissolved inorganic nitrogen (DIN) was determined in upland groundwater, riparian groundwater, and stream water of the Barro Branco catchment, Amazônas, Brazil. The ¹⁵N composition of organic nitrogen in riparian and upland leaf litter was also determined. The data for these waters could be divided into three groups: upland groundwater DIN predominately composed of NO₃ ^– with ¹⁵N values averaging 6.25 ± 0.9 riparian groundwater DIN primarily composed of NH₄ ⁺ with ¹⁵N values averaging 9.17 ± 1.0 and stream water DIN predominately composed of NO₃ ^– with ¹⁵N values averaging 4.52 ± 0.8 Nitrate samples taken from the stream source and from the stream adjacent to the groundwater transects showed a downstream increase in ¹⁵N from 1.0to 4.5 Leaf litter samples averaged 3.5 ± 1.2The observed patterns in isotopic composition, together with previously observed inorganic nitrogen species and concentration shifts between upland, riparian and stream waters, suggest that groundwater DIN is not the primary source of DIN to the stream. Instead, the isotopic data suggest that remineralization of organic nitrogen within the stream itself may be a major source of stream DIN, and that the majority of DIN entering the stream via groundwater flowpaths is removed at the riparian-stream interface.     

The Status and Characteristics of Eutrophication in the Yangtze River (Changjiang) Estuary and the Adjacent East China Sea, China

Eutrophication has become increasingly serious and noxious algal blooms have been of more frequent occurrence in the Yangtze River Estuary and in the adjacent East China Sea. In 2003 and 2004, four cruises were undertaken in three zones in the estuary and in the adjacent sea to investigate nitrate (NO₃–N), ammonium (NH₄–N), nitrite (NO₂–N), soluble reactive phosphorus (SRP), dissolved reactive silica (DRSi), dissolved oxygen (DO), phytoplankton chlorophyll a (Chl a) and suspended particulate matter (SPM). The highest concentrations of DIN (NO₃–N+NH₄–N+NO₂–N), SRP and DRSi were 131.6, 1.2 and 155.6 μM, respectively. The maximum Chl a concentration was 19.5 mg m⁻³ in spring. An analysis of historical and recent data revealed that in the last 40 years, nitrate and SRP concentrations increased from 11 to 97 μM and from 0.4 to 0.95 μM, respectively. From 1963 to 2004, N:P ratios also increased from 30–40 up to 150. In parallel with the N and P enrichment, a significant increase of Chl a was detected, Chl a maximum being 20 mg m⁻³, nearly four times higher than in the 1980s. In 2004, the mean DO concentration in bottom waters was 4.35 mg l⁻¹, much lower than in the 1980s. In comparison with other estuaries, the Yangtze River Estuary was characterized by high DIN and DRSi concentrations, with low SRP concentrations. Despite the higher nutrient concentrations, Chl a concentrations were lower in the inner estuary (Zones 1 and 2) than in the adjacent sea (Zone 3). Based on nutrient availability, SPM and hydrodynamics, we assumed that in Zones 1 and 2 phytoplankton growth was suppressed by high turbidity, large tidal amplitude and short residence time. Furthermore, in Zone 3 water stratification was also an important factor that resulted in a greater phytoplankton biomass and lower DO concentrations. Due to hydrodynamics and turbidity, the open sea was unexpectedly more sensitive to nutrient enrichment and related eutrophication processes.     

Subcellular and developmental studies of the tyrosyl protein sulfotransferase in rat brain

• 1.1. Tyrosyl protein sulfotransferase (TPS) activity in the newborn and mature rat brain was studied using the cholecystokinin derivative terbutyloxycarbonyl-Asp-Tyr-Met-Gly-Trp-Met-Asp-PheNH₂, BocCCK-8(ns), as the peptide substrate.
• 2.2. TPS activity was enriched 4 times in the microsomal and synaptic vesicular enriched fractions of rat cerebral cortex.
• 3.3. CCK-8 content, in the subcellular fractions and the peptide sulfation activity distribution was in accord with the hypothesis that tyrosyl protein sulfotransferase plays a key role in the maturation process of bioactive CCK.
• 4.4. TPS activity measured in membranes from newborn brain was 2.5 times higher than the activity observed in the mature brain membranes with a V_max = 0.83 ± 0.05 and 0.31 ± 0.02 respectively. The apparent K_M for the sulfate donor, 3'-phosphoadenosine 5'-phosphosulfate (PAPS), was similar, 94 ± 4 nM and 90 ± 6 nM and the k_M for the peptide substrate, BocCCK-8(ns), was 234 ± 16 μM and 160 ± 12 μM in the newborn and adult brain membranes respectively.
• 5.5. TPS activity reached normal mature values within 20 days of age.
• 6.6. These data support the idea that tyrosyl protein sulfation is an important process in the secretion mechanism and in the CCK maturation.