Macroalgal blooms on southeast Florida coral reefs: II. Cross-shelf discrimination of nitrogen sources indicates widespread assimilation of sewage nitrogen

Since 1990, coral reefs off southeast Florida have experienced an unprecedented succession of macroalgal blooms and invasions. To determine if anthropogenic land-based nitrogen (N) sources support these HABs, we collected macroalgal tissue for stable nitrogen isotope (δ¹⁵N) analysis at three spatially distinct depths ranging from the shallow subtidal to the shelf break (43 m) along seven transects from Jupiter to Deerfield Beach, Florida, USA. This sampling was initiated during a historically significant drought in the spring of 2001 when rainfall, stormwater runoff, and upwelling were at a minimum. The sampling was repeated in the summer wet season following significant stormwater runoff and during a strong upwelling event.Despite significant seasonal changes in N source availability, δ¹⁵N values did not vary between samplings. Collectively, δ¹⁵N values were significantly higher on inshore shallow reefs (+8.1‰) compared to mid (+6.1‰) and deep reefs (+6.7‰). Values were also elevated in the southern portion of the study area (e.g., Boca Raton, +8.5‰) where nearly 1.5 billion l/day of secondarily treated wastewater is discharged into the ocean via coastal outfalls. Codium isthmocladum, a macroalga that assimilates its nutrients entirely from the water column, was the dominant bloom species in the southern study area, where tissue δ¹⁵N values matched source values of nearby sewage outfalls. The northern study area was dominated by species of the genus Caulerpa, particularly the invasive Pacific native C. brachypus var. parvifolia, which are capable of accessing benthic nutrient sources (e.g., submarine groundwater discharge, SGD) by means of root-like rhizoids. The northern area does not have sewage outfalls but features a highly transmissive geologic zone where SGD may be enriched with septic tank leachate and effluent from municipal deep injection wells.Multiple lines of evidence supported the hypothesis that land-based sewage N was more important than upwelling as a N source to these HABs: (1) δ¹⁵N values were highest on shallow reefs and decreased with increasing depth, indicating land-based sources of enrichment; (2) elevated δ¹⁵N values occurred in these HABs during the dry season, prior to the onset of the summer upwelling; (3) elevated NH₄⁺ concentrations occur on these reefs during both upwelling and non-upwelling periods and are kinetically preferred by macroalgae compared to upwelled NO₃⁻. These findings provide a case study of a coupling between increasing anthropogenic activities and the development of macroalgal HABs, including invasive species that threaten economically important reef resources in southeast Florida.     

First occurrence of Cochlodinium blooms in Sabah, Malaysia

Harmful algal blooms (HABs) resulting in red discoloration of coastal waters in Sepanggar Bay, off Kota Kinabalu, Sabah, East Malaysia, were first observed in January 2005. The species responsible for the bloom, which was identified as Cochlodinium polykrikoides, coincided with fish mortalities in cage-cultures. Determinations of cell density between January 2005 and June 2006 showed two peaks that occurred in March–June 2005 and June 2006. Cell abundance reached a maximum value of 6 × 10⁶ cells L⁻¹ at the fish cage sampling station where the water quality was characterized by high NO₃–N and PO₄–P concentrations. These blooms persisted into August 2005, were not detected during the north–east monsoon season and occurred again in May 2006. Favorable temperature, salinity and nutrient concentrations, which were similar to those associated with other C. polykrikoides blooms in the Asia Pacific region, likely promoted the growth of this species. Identification of C. polykrikoides as the causative organism was based on light and scanning microscopy, and confirmed by partial 18S ribosomal DNA sequences of two strains isolated during the bloom event (GenBank accession numbers DQ915169 and DQ915170).     

Nutrient limitation of Microcystis aeruginosa in northern California Klamath River reservoirs

Nutrient limitations were investigated in Copco and Iron Gate Reservoirs, on the Klamath River in California, where blooms of the toxin-producing cyanobacterium Microcystis aeruginosa were first reported in 2005. Nutrient enrichment experiments conducted in situ in June and August, 2007 and 2008, determined responses in phytoplankton biomass, Microcystis abundance and microcystin concentration to additions of phosphorus and different forms of nitrogen (NH₄⁺, NO₃, and urea). Microcystis abundance was determined using quantitative PCR targeting the phycocyanin intergenic spacer cpcBA.Total phytoplankton biomass increased with additions of N both before and during Microcystis blooms, with no primary effects from P, suggesting overall N limitation for phytoplankton growth during the summer season. NH₄⁺ generally produced the greatest response in phytoplankton growth, while Microcystis abundance increased in response to all forms of N. Microcystis doubling time in the in situ experiments was 1.24–1.39 days when N was not limiting growth. The results from this study suggest availability of N during the summer is a key growth-limiting factor for the initiation and maintenance of toxic Microcystis blooms in Copco and Iron Gate Reservoirs in the Klamath River.     

Seasonal water quality of shallow and eutrophic Lake Pamvotis, Greece: implications for restoration

Lake Pamvotis is a moderately sized (22 km²) shallow (z _avg=4 m) lake with a polymictic stratification regime located in northwest Greece. The lake has undergone cultural eutrophication over the past 40 years and is currently eutrophic (annual averages of FRP=0.07 mg P l^-1, TP=0.11 mg P l^-1, NH₄ ⁺=0.25 mg N l^-1, NO₃ ^–=0.56 mg N l^-1). FRP and NH₄ ⁺ levels are correlated to external loading from streams during the winter and spring, and to internal loading during multi-day periods of summer stratification. Algal blooms occurred in summer (July–August green algae, August–September blue-green algae), autumn (October blue-green algae and diatoms), and winter (February diatoms), but not in the spring (March–June). The phytoplankton underwent brief periods of N- and P-limitation, though persistent low transparency (secchi depth of 60–80 cm) also suggests periods of light limitation. Rotifers counts were highest from mid-summer to early autumn whereas copepods were high in the spring and cladocerans were low in the summer. Removal of industrial and sewage point sources a decade ago resulted in a decrease in FRP. A phosphorus mass balance identified further reductions in external loading from the predominately agricultural catchment will decrease FRP levels further. The commercial fishery and lake hatchery also provides opportunities to control algal biomass through biomanipulation measures.     

Phytoplankton communities in navigation pool no. 7 of the Upper Mississippi River

A study of the phytoplankton community dynamics in Navigation Pool No. 7 of the Upper Mississippi River was conducted from May through October, 1982. The objectives of this study were to estimate total standing crops, determine the taxonomic composition and examine the seasonal succession of the phytoplankton community. Four sampling sites were established: two in Lake Onalaska, a large backwater lake on the Wisconsin side of the main channel; one in the main channel near Dakota, Minnesota; and one in the main channel just upstream from Lock and Dam No. 7.The phytoplankton communities at all sampling sites were dominated by diatoms except during July and August when a bloom of blue-green algae was observed. The dominant diatoms from May through mid-July were Melosira italica, Stephanodiscus niagarae, Stephanodiscus hantzschii, Stephanodiscus astrea, and Synedra ulna. Aphanizomenon flos-aquae and Microcystis aeruginosa were the most prevalent blue-green algae during the mid-summer bloom. The diatoms Melosira italica and Melosira granulata were dominant in September and October. Lesser amounts of green algae, cryptomonads and euglenoids were also observed at various times of the sampling period.Total standing crops based on cell volume were usually greatest at the Lock and Dam No. 7 site. The maximum standing crop (10.4 mm³ 1^–1) was observed at the Lock and Dam No. 7 site on 4 September; the minimum standing crop (0.4 mm³ 1^–1) was observed at the eastern Lake Onalaska site on the same date. Concentrations of nitrogen, phosphorus, and silica remained at high levels throughout the study period and did not appear to limit phytoplankton standing crops.     

Does uptake of Alexandrium fundyense cysts contribute to the levels of PSP toxin found in the sea scallop, Placopecten magellanicus?

Atlantic sea scallops, Placopecten magellanicus, in most areas of the Bay of Fundy, New Brunswick, Canada, have year-round concentrations of paralytic shellfish posioning (PSP) toxins greater than the regulatory concentration of 80 μg STX eq. 100 g⁻¹ wet weight. Scallops (mean shell height of 10.7 cm, age 3–5 years) were collected by SCUBA and individually tagged near Parker Island, Bay of Fundy. Half were hung 2 m below the low tide water level and the remainder were placed on the bottom (11 m depth at low tide) under the scallops held at 2 m. Scallop, water and sediment samples were collected monthly for determination of concentrations of PSP toxins and Alexandrium fundyense.In October, 1993, mean concentrations of PSP toxins in digestive gland, and mantle were 3205 and 1018 μg STX eq. 100 g⁻¹ wet weight, respectively. Eight months later (June 1994), PSP concentrations in digestive glands from the surface and bottom had declined to 504 and 682 μg STX eq. 100 g⁻¹ wet weight, respectively, whereas those in the mantle had declined to 802 and 681 μg STX eq. 100 g⁻¹ wet weight. During July 1994, A. fundyense concentrations observed at Parker Island and offshore were 320 cells l⁻¹ and 14,200 cells l⁻¹, respectively. Subsequently, toxin concentrations in surface and bottom scallop digestive glands increased to 12,720 and 11,408 μg STX eq. 100 g⁻¹ wet weight, whereas concentrations in mantles increased to 2126 and 1748 μg STX eq. 100 g⁻¹ wet weight, respectively. Concentrations of PSP toxins in these tissues in October 1994 were similar to those measured in October 1993. Concentrations of PSP toxin were less than the regulatory concentration in the gonads and non-detectable in adductor muscles of all scallops sampled.There were no statistically significant differences in profiles for uptake and depuration of PSP toxins in scallops held at the surface compared to those from bottom, suggesting that A. fundyense cysts at the concentrations found in the sediment (45 cysts cm⁻³) did not contribute significantly to the year-round presence of PSP toxins within scallop tissues. The year-round occurrence of PSP toxin is probably due to accumulation during summer blooms followed by a very slow rate of depuration.     

Resource use by salmonids in riverine,lacustrine and marine environments: Evidence from stable isotope analysis

Synopsis We measured stable isotope ratios (δ¹³C and δ¹⁵N) of invertebrates, Atlantic salmon, Salmo salar, and brook trout, Salvelinus fontinalis, in three distinct freshwater environments (headwater tributary, ultra-oligotrophic lake, and main-stem river) in the Western Brook system, Newfoundland, Canada. Large differences in the stable carbon signatures of invertebrates allowed the identification of organic matter assimilation from each environment by resident parr and migrating smolts. Brook trout captured in the headwater tributary in June had a carbon signature characteristic of the tributary, while those collected in August had enriched ¹³C (maximum = −15.6‰) and ¹⁵N (maximum = 12.8‰) values. These enriched carbon and nitrogen signatures were indicative of foraging at sea. There was a low correlation between δ¹³C and δ¹⁵N (r² = 0.198) for individual fish that was likely due to the confounding influence of trout feeding in the lake and the lower main-stem of the river, where δ¹³C of food sources was high but δ¹⁵N was low. Smolts emigrating from Western Brook Pond where they had been foraging (based on lacustrine carbon signatures) were significantly larger than those emigrating from a nursery brook and the main river in the same basin, despite having the same median age. These results suggest better growth opportunities in the lake environment. Trout fork length was positively correlated with δ¹³C and δ¹⁵N, demonstrating that larger individuals had been feeding outside the brook. These results support previous studies that found increased growth potential for salmonids in lacustrine and marine environments, and further, indicate possible adaptive advantages for salmonid movement away from natal brooks.     

Effect of temperature on light-limited growth of the harmful diatom Eucampia zodiacus Ehrenberg, a causative organism in the discoloration of Porphyra thalli

The diatom Eucampia zodiacus Ehrenberg is one of the harmful diatoms which indirectly cause, through nutrient depletion, discoloration of Porphyra thalli. The effect of temperature on light-limited growth of E. zodiacus was examined at 13 irradiance levels (5–350 μmol m⁻² s⁻¹) in combination with five temperatures (8.0–25.0 °C). The results showed that all the parameters of growth-irradiance curves, such as the maximum growth rate (μ_m), half saturation constant (K_s), threshold value of irradiance (I₀) and saturation irradiance for growth (S), increased with increasing temperature. On the basis of the relationship between temperature and growth-irradiance curves and seasonal fluctuation of the light environment in Harima-Nada, the effect of irradiance on the population dynamics of E. zodiacus during the period from October to March was evaluated using two indices, depth of the threshold irradiance for growth (D_t) and depth where a half of its maximum growth rate is attained (D_k). D_t and D_k remained almost stable from October to December, but gradually increased in early March. This indicates that the range of depth at which E. zodiacus was able to grow increased markedly in early spring when E. zodiacus blooms in Harima-Nada. As the vegetative cells of E. zodiacus tend to distribute in relatively deeper water layers, where growth is limited by irradiance, the increase in the depth range over which E. zodiacus is able to grow is concluded to be an important factor allowing development of its blooms.     

Red tide blooms of Cochlodinium polykrikoides in a coastal cove

Successive blooms of the dinoflagellate Cochlodinium polykrikoides occurred in Pettaquamscutt Cove, RI, persisting from September through December 1980 and again from April through October 1981. Cell densities varied from <100 cells L⁻¹ at the onset of the bloom and reached a maximum density exceeding 3.4 × 10⁶ cells L⁻¹ during the summer of 1981. The bloom was mainly restricted to the mid to inner region of this shallow cove with greatest concentrations localized in surface waters of the southwestern region during summer/fall periods of both years. Highly motile cells consisting of single, double and multiple cell zooids were found as chains of 4 and 8 cells restricted to the late August/September periods. The highest cell densities occurred during periods when annual temperatures were between 19 and 28 °C and salinities between 25 and 30. A major nutrient source for the cove was Crying Brook, located at the innermost region at the head of the cove. Inorganic nitrogen (NH₃ and NO₂ + NO₃) from the brook was continually detectable throughout the study with maximum values of 57.5 and 82.5 μmol L⁻¹, respectively. Phosphate (PO₄-P) was always present in the source waters and rarely <0.5 μmol L⁻¹; silicate always exceeded 30 μmol L⁻¹ with maximum concentrations reaching 226 μmol L⁻¹. Chlorophyll a and ATP concentrations during the blooms varied directly with cell densities. Maximum Chl a levels were 218 mg m⁻³ and ATP-carbon was >20 g C m⁻³. Primary production by the dinoflagellate-dominated community during the bloom varied between 4.3 and 0.07 g C m⁻³ d⁻¹. Percent carbon turnover calculated from primary production values and ATP-carbon varied from 6 to 129% d⁻¹. The dinoflagellates dominated the entire summer period; other flagellates and diatoms were present in lesser amounts. A combination of low washout rate due to the cove dynamics, active growth, and life cycles involving cysts allowed C. polykrikoides to maintain recurrent bloom populations in this area.     

δC, δN, carbon, nitrogen and phosphorus as indicators of plant ecophysiology and organic matter pathways in Everglades deep slough, Florida, USA

Historically, the Florida Everglades was characterized by a corrugated landscape of shorter hydroperiod, elevated sawgrass (Cladium jamaicense) ridges and longer hydroperiod, deep water slough communities. Drainage and compartmentalization of the Everglades have fundamentally altered this pattern, and sawgrass ridge communities have expanded at the expense of deep water slough communities throughout much of the landscape. In this study we provide a simple isotopic and nutrient characterization of major components of the slough ecosystem to elucidate physiological and nutrient differences among species and to suggest pathways for organic matter decomposition that contribute to peat development in deep water sloughs. We examined carbon (C) and nitrogen (N) isotopes and C, N and phosphorus (P) concentrations of the floating-leaved macrophytes Nymphaea odorata and Nymphoides aquatica, the emergent macrophyte Eleocharis elongata, and the submerged species Utricularia foliosa and Utricularia purpurea, as well as soil and flocculent material from the southern Water Conservation Area 3-A. Flocculent material and soils had the highest N content (4.5 ± 0.2%) and U. foliosa and N. odorata had the highest P content (0.13 ± 0.01% to 0.12 ± 0.01%). The range for δ¹⁵N average ± SE values was 5.81 ± 0.29‰ (U. foliosa) to −1.84 ± 0.63‰ (N. odorata), while the range for δ¹³C values was −23.83 ± 0.12‰ (N. odorata) to −29.28 ± 0.34‰ (U. purpurea). Differences of up to 10‰ in C isotopic values of U. foliosa and N. odorata suggest fundamental physiological differences between these species. Along a degradation continuum, enrichment of ¹³C and ¹⁵N and extent of decomposition was negatively related to phosphorus concentrations. A two end-member ¹³C mixing model suggested that Utricularia species were the primary organic source for flocculent materials, whereas organic matter derived from root decomposition of N. odorata contributed to the progressively enriched δ¹³C values found with depth in soils. These results illustrate the fundamentally important roles of Nymphaea and Utricularia species in ecosystem dynamics of deep water sloughs.     

Occurrence and toxicity of an Anabaena bloom in a tropical reservoir (Southeast Brazil)

Potentially toxic cyanobacterial blooms are becoming common in the Brazilian reservoirs in all regions of the country. During October 2004, a dense bloom of cyanobacteria occurred in the Monjolinho Reservoir (São Carlos, São Paulo State, Brazil) and a significant amount of cyanobacterial material accumulated on the water surface. Phytoplankton analysis showed that the main species in this bloom were Anabaena circinalis and Anabaena spiroides. Cladoceran (Ceriodaphnia dubia and Ceriodaphnia silvestrii) and mouse bioassays were performed to detect toxic products in extracts of the natural samples collected at the three different dates during in short period. To prepare the extracts, freeze-dried cells were dispersed in distilled water and subjected to repeated freeze/thaw cycles and sonication and centrifuging processes. Crude extracts were toxic both to cladocerans (LC₅₀ 94–406 mg freeze-dried cells L⁻¹) and mice (indicative LD₅₀ 297–445 mg freeze-dried cells kg⁻¹) and the toxicity of the bloom increased for cladocerans during the occurrence of the bloom. Toxin analysis by ELISA revealed that microcystin (MC) was found in the water of the reservoir (concentrations ranging from 28 to 45 μg L⁻¹). In addition, microcystin was also found in freeze-dried cyanobacteria cells with concentrations ranging from 138 to 223 μg g⁻¹. On the other hand, neurotoxins (saxitoxin and gonyautoxin) were not detected in any of the natural samples by HPLC. Signs of toxicity in mice did not indicate whether the bloom samples were predominantly hepatotoxic or neurotoxic. It is known that natural Anabaena blooms can contain other toxic compounds besides microcystins and neurotoxins such as lipopolysaccharides or other toxins not identified or known. Methods of detecting cyanotoxins used in this study were insufficient to clarify the toxicological features of Anabaena bloom and indicated that other methods should be investigated.     

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.     

Relative Uptake of Urea and Ammonium by Dinoflagellates or Cyanobacteria in Shrimp Mesocosms

The relative role of the organic nitrogen source, urea, versus ammonium as a nitrogen source for two species of dinoflagellates was compared with one species of cyanobacteria. Experiments were conducted opportunistically in nutrient-rich marine water during blooms of 34either cyanobacteria or dinoflagellates in outdoor mesocosms. These replicate mesocosms, which were stocked with shrimp fed high-protein formulated feeds, contained high biomasses of phytoplankton (mean chlorophyll a concentrations, 439.2–811.2 μg l⁻¹). ¹⁵N-urea and ammonium uptake rates for dinoflagellate-dominated blooms (Gymnodinium pulchellum-complex (Larsen), Karlodinium micrum (Larsen) (Dinophyceae)) were compared with blooms of the cyanobacterium, Romeria sp. (Cyanophyceae) in mesocosms with mean urea and ammonium concentrations ranging from 2.32 to 3.24 μM, and 7.39 to 64.85 μM, respectively. Urea uptake rates were significantly (p < 0.005) lower than ammonium uptake rates irrespective of which algal species dominated the bloom. Additionally urea uptake rates were not significantly higher in G. pulchellum-complex or K. micrum-dominated blooms than in Romeria sp. blooms. These results suggest that G pulchellum complex and K. micrum may not be gaining a competitive advantage in waters high in dissolved organic matter simply by preferentially utilizing urea. The periodic dominance of these species in highly organic environments, such as shrimp ponds, is likely to have a more complex explanation.     

Growth of dinoflagellates, Ceratium furca and Ceratium fusus in Sagami Bay, Japan: The role of temperature, light intensity and photoperiod

Seasonal changes of field populations and growth rates of two dinoflagellates, Ceratium furca and Ceratium fusus, were examined in the temperate coastal water of Sagami Bay, Japan. Weekly field sampling was conducted from August 2002 to August 2003, and laboratory experiments were also carried out to investigate effects of temperature, irradiance and photoperiod on the growth rates of these two Ceratium species. In the field, the abundances of both species increased significantly from April to August 2003, were gradually decreased from November 2002 and were not observed in January 2003. C. fusus was able to increase at lower temperatures in February 2003 compared to C. furca. In the laboratory, the two species did not grow at <10 °C or >32 °C. The highest specific growth rate of C. furca was 0.72 d⁻¹ at 24 °C and 600 μmol m⁻² s⁻¹. Optimum growth rates (>0.4 d⁻¹) of C. furca were observed at temperatures from 18 to 28 °C and at irradiances from 216 to 796 μmol m⁻² s⁻¹. The highest growth rate of C. fusus was 0.56 d⁻¹ at 26 °C and 216 μmol m⁻² s⁻¹. Optimum growth rates of C. fusus were observed at the same irradiance rage of C. furca, whereas optimum temperature range was narrower (26–28 °C). The growth curves of both species indicated saturation of the growth rates when light intensity was above 216 μmol m⁻² s⁻¹, and did not show photoinhibition at irradiances up to 796 μmol m⁻² s⁻¹. The specific growth rates of both Ceratium species were clearly decreased at L:D = 10:14 relative to those at L:D = 14:10 and L:D = 12:12. The present study indicates the two Ceratium species can adapt to a wide range of temperature and irradiance.     

Inorganic nutrient and oxygen fluxes across the sediment–water interface in the inshore macrophyte areas of a shallow estuary (Lake Illawarra,Australia)

Rates of inorganic nutrient and oxygen fluxes, and gross community primary productivity were investigated using incubated cores in July, August and September 2001, in a seagrass meadow of Lake Illawarra, a barrier estuary in New South Wales, Australia. The results indicated that rates of gross primary productivity were high, varying from C = 0.62 to 1.89 g m^–2 d^–1; low P/R ratios of 0.28–0.48 define the system as heterotrophic and indicate that more carbon is respired than is produced. In order to determine the effect of macroalgae on O₂ and nutrient fluxes, measurements were also conducted on cores from which the macroalgae had been removed. The results showed that the O₂ fluxes during light incubations were significantly lower in the cores without macroalgae (P<0.01), indicating that macroalgae could be a significant contributor to the primary production in the lake. In general, nutrient fluxes showed a typical diurnal variation with an efflux from sediments in the dark and a reduced efflux (or uptake) in the light. Dissolved inorganic nitrogen (NO₂ ^–+ NO₃ ^–+NH₄ ⁺) net fluxes were directed from the sediments towards the water column and dominated by the NH₄ ⁺ fluxes (>80%). NO₂ ^–+ NO₃ ^– and o-P fluxes were always very low during the sampling period. The increasing tendency of net nutrient effluxes, especially NH₄ ⁺ from July to September, is consistent with the increase of the water temperature and seagrass biomasses. However, in September, significantly lower light, dark and net NH₄ ⁺ effluxes were found in the cores with macroalgae (SA-sediments) compared with the cores without macroalgae (S-sediments). These results support the hypothesis that actively-growing dense macroalgal mats (i.e., algal blooms in September) may act as a filter reducing the flux of nutrients to the water column.     

Nitrogenous preference of toxigenic Pseudo-nitzschia australis (Bacillariophyceae) from field and laboratory experiments

Field and laboratory experiments were designed to determine the differential growth and toxin response to inorganic and organic nitrogen additions in Pseudo-nitzschia spp. Nitrogen enrichments of 50 μM nitrate (KNO₃), 10 μM ammonium (NH₄Cl), 20 μM urea and a control (no addition) were carried out in separate carboys with seawater collected from the mouth of the San Francisco Bay (Bolinas Bay), an area characterized by high concentrations of macronutrients and iron. All treatments showed significant increases in biomass, with chlorophyll a peaking on days 4–5 for all treatments except urea, which maintained exponential growth through the termination of the experiment. Pseudo-nitzschia australis Frenguelli abundance was 10³ cells l⁻¹ at the start of the experiment and increased by an order of magnitude by day 2. Particulate domoic acid (pDA) was initially low but detectable (0.15 μg l⁻¹), and increased throughout exponential and stationary phases across all treatments. At the termination of the experiment, the urea treatment produced more than double the amount of pDA (9.39 μg l⁻¹) than that produced by the nitrate treatment (4.26 μg l⁻¹) and triple that of the control and ammonium treatments (1.36 μg l⁻¹ and 2.64 μg l⁻¹, respectively). The mean specific growth rates, calculated from increases in chlorophyll a and from cellular abundance of P. australis, were statistically similar across all treatments.These field results confirmed laboratory experiments conducted with a P. australis strain isolated from Monterey Bay, CA (isolate AU221-a) grown in artificial seawater enriched with 50 μM nitrate, 50 μM ammonium or 25 μM of urea as the sole nitrogen source. The exponential growth rate of P. australis was significantly slower for cells grown on urea (ca. 0.5 day⁻¹) compared to the cells grown on either nitrate or ammonium (ca. 0.9 day⁻¹). However the urea-grown cells produced more particulate and dissolved domoic acid (DA) than the ammonium- or nitrate-grown cells. The field and laboratory experiments demonstrate that P. australis is able to grow effectively on urea as the primary source of nitrogen and produced more pDA when grown on urea in both natural assemblages and unialgal cultures. These results suggest that the influence of urea from coastal runoff may prove to be more important in the development or maintenance of toxic blooms than previously thought, and that the source of nitrogen may be a determining factor in the relative toxicity of west coast blooms of P. australis.     

Microcystin production by cyanobacteria in the Mwanza Gulf (Lake Victoria,Tanzania)

In order to investigate the potential for microcystin (MC) production by cyanobacteria in the Mwanza Gulf (Lake Victoria, Tanzania), nutrients, phytoplankton and microcystins were sampled inshore (3 m depth) and offshore (18 m depth) from May to August 2002. Significant differences in soluble reactive phosphorus (SRP) and nitrate concentrations between offshore and inshore indicated eutrophication via terrestrial run-off. Though the concentrations of SRP and nitrate ranged between 36–127 and 35–726 μg l ⁻¹ each, the phytoplankton biovolume was generally low. The phytoplankton community was dominated by diatoms (Nitzschia acicularis), a number of cyanobacterial species (Aphanocapsa sp., Anabaena sp., Planktolyngbya spp., Microcystis sp.) and cryptomonads. The water column was completely mixed and Nitzschiapeaked in abundance during July. All cyanobacteria were low in abundance during the entire study period (0.1–1.6 mm ³ l ⁻¹). Microcystins were analysed using high performance liquid chromatography coupled with diode array detection High Performance Liquid Chromatography with Diode Array Detection (HPLC-DAD) and in most samples no microcystins were detected. The highest concentration of [Asp ³]-MC-RR was found in open water at the surface on July 2nd, 2002 (1 μg l ⁻¹). MC concentrations did not pose a potential health risk in the Mwanza Gulf during the study period, however, it is possible that the period of higher cyanobacterial biovolumes has been missed during the sampling period of this study.     

Potential impacts of brown tide, Aureococcus anophagefferens, on juvenile hard clams, Mercenaria mercenaria, in the Coastal Bays of Maryland, USA

The rate of growth of juvenile hard clams, Mercenaria mercenaria, was studied in the Coastal Bays of Maryland during an outbreak of the brown tide, Aureococcus anophagefferens. Brown tide dominated the plankton community during the month of June 2002, with cell densities at several sites reaching category 3 (>200,000 cells ml⁻¹) levels. Temperatures during the bloom were 18.6–27.5 °C. Nutrient conditions preceding and during the bloom were conducive for the proliferation of A. anophagefferens: while inorganic nitrogen and phosphorus were <1 μg at N or P l⁻¹, urea was elevated during bloom development. Organic nitrogen, phosphorus and carbon were in the range of levels observed in previous brown tide blooms and increased following the collapse of the bloom. Growth rates of juvenile clams were significantly lower during the period of the brown tide bloom than following its collapse. Growth rates of M. mercenaria were found to be negatively impacted at brown tide densities as low as 20,000 cells ml⁻¹, or category 1 levels. The low growth rates of M. mercenaria could not be explained by temperature, as the lowest growth rates were found when water temperatures were at levels previously found to be optimal for growth.     

Dynamics of transparent exopolymeric particles (TEP) production by Phaeocystis globosa under N- or P-limitation: a controlling factor of the retention/export balance

The concentration of transparent exopolymeric particles (TEP) was monitored during Phaeocystis globosa blooms that developed in mesocosms under different initial N:P ratios (from N- to P-limited conditions). TEP concentration was measured using the microscopic (TEP_micro, ppm) and the colorimetric (TEP_color, Xanthan equiv. L⁻¹) methods. TEP concentrations varied from 5 to >75 ppm and from 60 to >1500 μg Xanthan equiv. L⁻¹, and were relatively low until the mesocosms reached nutrient (either N or P) depletion and then increased abruptly. From the TEP_micro versus TEP_color concentrations comparison and from their relation to chlorophyll a concentrations, two phases for the dynamics of TEP production were identified: (1) production through active release of precursors during the growth phase of P. globosa — defined as TEP₁ — and their integration into the TEP pool through coagulation processes; (2) release of large TEP from the mucilaginous matrix of P. globosa colonies subsequent to disruption caused by nutrient depletion — defined as TEP₂ — and their direct integration into the TEP pool outside the constraint of coagulation. The formation of a multiorigin TEP pool during P. globosa blooms may have implications for the fate of the blooms, due to difference in TEP bioreactivity according to their source and to difference in timing and intensity of TEP₁ versus TEP₂ production according to N- or P-depletion. For P. globosa blooms developing under N-limiting conditions, the transition from the first source (i.e. TEP₁) to the second one (i.e. TEP₂) was a slow and continuous process. In contrast, the P. globosa bloom developing under P-limiting conditions showed the sudden formation of heavy mucous aggregates when P became depleted, that may have been caused by a massive release of TEP₂. Our study suggests that the nutrient regime may control the export vs. retention balance during P. globosa blooms, via production of a multiorigin TEP pool.     

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.