Diel periodicity of photosynthesis in marine phytoplankton

Short-term changes in photosynthesis were documented for 17 of 24 marine phytoplankton species, representing a range of taxonomic groups. Periodicity in phytoplankton photosynthesis on light-dark cycles (diel periodicity) was widespread but not universal for the species studied. The centric diatoms Lauderia borealis, Ditylum brightwellii, Stephanopyxis turris, Coscinodiscus rex, Chaetoceros gracile, and Biddulphia mobiliensis had strong diel periodicity in photosynthetic capacity (P _max). Amplitudes of the daily variations ranged from 2.9 to >50, with maxima in the morning or near midday, and with minima during the dark period, and these variations were not dependent on changes in cell pigmentation. There was some evidence for sustained photosynthetic periodicity in constant conditions in several diatoms, and an endogenous rhythm may have been present. The photosynthesis-irradiance (P-I) relationship was time-dependent for representative marine diatoms, with both the initial slope () and the asymptote (P _max) of P-I curves exhibiting significant synchronous diel oscillations. Moreover, detailed studies of the amplitude and timing of photosynthetic periodicity for the diatoms L. borealis and D. brightwellii demonstrated large temporal variations in photosynthesis with morning maxima. These P-I oscillations are discussed with reference to models of primary production which use the relationship between photosynthesis and light as a component of predictive equations for phytoplankton growth in the sea.     

Diel variation in chlorophyll a,carbohydrate and protein content of the marine diatom Skeletonema costatum

Skeletonema costatum (Greville) Cleve isolated from Narragansett Bay, USA, was incubated at 3 light intensities (ca. 0.008, 0.040 and 0.075 ly min^-1) under a 12 h light: 12 h dark (12L:12D) photoperiod at 2°, 10° and 20°C. Cellular chlorophyll a increased at intensities less than ca. 0.040 ly min^-1; increases occured within one photoperiod at temperatures above 10°C. Cellular carbohydrate increased with light intensity at all temperatures; increases during the photophase were due to net production of the dilute acid-soluble fraction. Cellular protein increased during the photoperiod at 10° and 20°C; there was little difference in cellular protein among all cultures after one photoperiod. The rate at which cellular chlorophyll a increased in response to a decrease in light suggests that diel variation in cellular chlorophyll a is temperature-dependent in S. costatum. Protein: carbohydrate ratios ranged from ca. 0.5 to 2.0 over a diel cycle; ratios increased at lower intensities and higher temperatures. The diel range in protein:carbohydrate ratios equals that in cultures developing nitrogen deficiency; thus, use of this ratio as an index to phytoplankton physiological state must account for diel light effects.     

Pools of chlorophyll and live planktonic diatoms in aphotic marine sediments

Chlorophyll a and numbers of live pelagic diatoms were recorded from sediment depth profiles at 11 stations in the oligotrophic Øresund, Denmark, in late-June. Extraction efficiency of chlorophyll a analysed fluorometrically did not differ significantly between paired samples of frozen-thawed and fresh sediment. The depth profiles of chlorophyll a could be explained by a diagenetic model involving two different chlorophyll pools: one reactive pool declining exponentially with core depth, and one non-reactive pool, of about 1 µg Chl ml^-1 wet sediment, being constant with depth. The number of live diatoms, quantified by the dilution-extinction method, and expressed in terms of most probable number (MPN), declined from an average of about 300,000 g^-1 in the surface sediment to zero values at a depth of 13 cm. The number of live cells was significantly correlated with the sediment chlorophyll a, and the profiles of live cells as well as reactive chlorophyll followed the same exponential decline with core depth, suggesting that the main source of chlorophyll in the sediment was live pelagic diatoms. Taxonomic composition of diatoms in the sediment, dominated by the pelagic genera Chaetoceros, Thalassiosira and Skeletonema, matched the species composition in the water column 3 months earlier during the spring bloom. Regular recordings of the phytoplankton community in the water column showed that only these specific bloom species could be the source of the sediment content of diatoms and chlorophyll a. Further, the ratios between live cells and chlorophyll a were similar in the sediment and in the spring bloom. A conservative estimate of depth-integrated pools of diatoms in the sediment suggested that about 44% of the total phytoplankton biomass during the spring bloom was still present as live cells in the sediment after 3 months. This indicates that the spring bloom input to the sediment is not degraded immediately by the benthic fauna.     

Dark survival of autotrophic,planktonic marine diatoms

A general ecological problem is considered: how long can a photoautotrophic microalga, incapable of producing a resting spore (stage), retain its viability in the dark following removal from the euphotic zone? Nine coastal diatoms, including some capable of producing resting spores, were kept in the dark for 90 days at 15°C, and their growth (viability) checked at periodic intervals upon reillumination. Seven of the 9 diatoms retained their viability for 90 days; generation time of illuminated cells then ranged from 2.5 to 10 days. Skeletonema costatum survived only 7 weeks of darkness. Based on the present and published observations, dark survival of this species is inversely related to temperature; it survives at least 24 weeks at 2°C, and from 1 to 4 weeks at 20°C. None of the species was observed to grow in the dark. The effects of temperature and light on dark survival, and of darkness on the chemical composition and photosynthesis following reillumination as reported in the scattered literature are evaluated. Together with the present observations, it is suggested that dark survival of photoautotrophic microalgae: (1) varies between species; (2) may be temperature dependent in some species, as in S. costatum; (3) may be prolonged by periodic illumination at subcompensation intensities for photosynthesis, as shown in Dunaliella tertiolecta. The potential ecological significance of these findings is also considered, should these in vitro results apply to natural populations.     

Significance of cellular nitrate content in natural populations of marine phytoplankton growing in shipboard cultures

Phytoplankton intracellular nitrate concentrations have been monitored in a 56-h experiment on a shipboard culture of surface sea water from an upwelling region. These measurements were related to parameters of biomass (particulate nitrogen) and nitrate assimilation using the ¹⁵N isotope technique and the nitrate reducase (NR) assay. The procedure for measuring cellular nitrate concentrations is described. This parameter exhibited diurnal variations, ranging from 3.1 to 20.6 ng-at nitrate per g-at particulate nitrogen, and could be correlated positively with NR activity. Nitrogen budgets show that NR activity represents only 12% of nitrate incorporation in organic phytoplankton material when nitrate is available in the sea water. However, upon depletion of the environmental nitrate (zero uptake), NR activity can fully account for the decrease of internal nitrate. From the results, it seems that internal nitrate content is a better index of nitrate consumption by marine phytoplankton than the external concentration of nitrate-nitrogen.     

Growth of marine planktonic diatoms on inorganic and organic nitrogen

A study was conducted to determine if coastal diatoms from eutrophic waters (Werribee, Port Phillip Bay, Australia) are able to grow better than diatoms from oligotrophic waters (Bass Strait, Australia) on organic nitrogen compounds as their principal nitrogen sources. Eight clones of marine planktonic diatoms, belonging to 5 species (Skeletonema costatum, Asterionella japonica, Nitzschia closterium, Coscinodiscus sp., and Fragilaria sp.), were incubated with inorganic (either nitrate or ammonia) or organic (either urea, uric acid, alanine, aspartic acid, glutamic acid, glycine, serine, threonine, or valine) nitrogen sources and growth response was measured under high and low light intensities. All clones grew well on the organic as well as thorganic nitrogen compounds under both light regimes. Intraspecific differences were not great, as no appreciable difference was noted between clones from oligotrophic and eutrophic waters. The two negatively-charged amino acids, aspartic and glutamic acids, were somewhat less effective in supporting growth of some clones than were the other amino acids. Virtually all of the dissolved organic nitrogen (DON) compounds tested were utilizable for algal growth. Further, all clones appeared able to utilize at least some natural DON (uncharacterized) for cell division; in 1980, DON represented 97% of total nitrogen in Bass Strait and 83% of total nitrogen in Werribee waters. The results are consistent with previous findings on algal utilization of certain DON compounds and indicate comparable abilities of cells from oligotrophic and eutrophic coastal waters to assimilate these nutrients.Publication No. 307 in the Ministry for Conservation, Victoria, Environmental Studies Series     

Enhanced primary production and nutrient regeneration within aggregated marine diatoms

Blooms of chain-forming diatoms often terminate with the mass flocculation and subsequent settlement of cells from the nutrient-depleted euphotic zone. While mass diatom aggregation has been suggested as an adaptive mechanism for placing resting spores in the deep sea, we hypothesized that aggregation may confer an immediate adaptive advantage to the associated diatoms as well. We tested this hypothesis by comparing the photosynthetic activity, pigment composition and nutrient-uptake rates of aggregated and suspended diatoms over time. Diatom aggregates were collected by SCUBA divers in the Santa Barbara Channel (34°23N; 119°50W) on 4 March 1987 and monitored for 9 d in the laboratory. Diatom aggregates sustained chlorophyll a-specific primary production rates two to nine times higher than those of freely suspended diatoms from the surrounding seawater. The timing of maximum productivity was strongly correlated with the appearance of remineralized ammonia within the aggregates. Chlorophyll a-specific nitrate-uptake rates were routinely three to nine times lower in diatom aggregates than in the surrounding seawater. Primary production and pigment concentrations of diatom aggregates aged in situ displayed changes similar to those observed in the laboratory. These results suggest that diatoms associated with aggregates maintain higher photosynthetic rates than freely suspended diatoms by efficiently exploiting remineralized ammonia within the aggregate microenvironment, in preference to external nitrate sources. The enhanced nutrient environment within aggregates may be important for understanding the adaptive significance of the mass flocculation of diatom blooms.     

Quantitative evidence concerning the stabilization of sediments by marine benthic diatoms

Six species of benthic diatoms and a natural benthic diatom community were cultured in flasks on a variety of sediments. Diatom species which secreted large quantities of mucilage were effective sediment stabilizers. These mucilage-secreting species significantly reduced resuspension and retarded laminar flow of the sediments when the culture flasks were agitated. Diatom species which secreted little or no mucilage were not effective sediment stabilizers. These non-mucilage-secreting species did not significantly effect resuspension or laminar flow of the sediments when the culture flasks were agitated. A sediment stabilizing mechanism based on the secretion of mucilage by pennate benthic diatoms is proposed. The effect such a process may have on distributional patterns of benthic invertebrates in areas where extensive diatom or other microalgal films occur is discussed.This work was supported by the Belle W. Baruch Foundation, and is Contribution No. 68 of the Belle W. Baruch Institute for Marine Biology and Coastal Research.     

Light response curve methodology and possible implications in the application of chlorophyll fluorescence to benthic diatoms

Chlorophyll a fluorescence has been increasingly applied to benthic microalgae, especially diatoms, for measurements of electron transport rate (ETR) and construction of rapid light response curves (RLCs) for the determination of photophysiological parameters [mainly the maximum relative ETR (rETR_max), the light saturation coefficient (E _k) and the maximum light use coefficient (α)]. Various problems with the estimation of ETR from the microphytobenthos have been identified, especially in situ. This study further examined the effects of light history of the cells and light dose accumulation during RLCs on the fluorescence measurements of ETR using the benthic diatom Navicula phyllepta. RLCs failed to saturate when using incremental increases in irradiance, however, curves with decreasing irradiance did saturate. Patterns indicating photoacclimation in response to light histories were observed, with higher rETR_max and E _k, and lower α, at high light compared to low light. However, these differences could be negated by increasing the RLC irradiance duration from 30 to 60 s. It is suggested that problems arose as a result of rapid fluorescence variations due to ubiquinone (Q_A) oxidation and non-photochemical chlorophyll fluorescence quenching (NPQ) which depended upon the light history of the cells and the RLCs accumulated light dose. Also, RLCs with irradiance duration of 10 s were shown to have a high level of error possibly specific to the fluorimeter programming. It is suggested that RLCs, using a Diving-PAM fluorimeter on benthic diatoms, should be run using decreasing irradiance steps of 30 s duration.     

Diel changes in gut-cell morphology and digestive activity of the marine copepod Acartia tonsa

Morphometric measurements of the gut cells of the marine calanoid copepod Acartia tonsa were made over a 24 h diel cycle to test the hypothesis that feeding rhythms in this copepod are limited by the cycles of blishter-like cells (B-cells) in the gut. Copepods were collected from Long Island Sound, New York, USA, in June 1991. Experimental treatments included low (1 g chlorophyll al^-1) and high (10 g chlorophyll al^-1) food concentrations. Copepods were fixed and embedded in Epon at six time intervals over the 24 h period, and semi-thin sections through the length of Midgut Zone II were analyzed for gut-cell area and vacuole area, with area measurements integrated to yield estimates of gut-cell volume and vacuole volume. Concurrent measurements of gut fluorescence, a measure of gut fullness, and digestive enzyme activities also were made. A diel cycle in gut fluorescence was observed, most notably in the low-food treatment, which exhibited a 3-fold increase in gut pigments. There was little consistent change in digestive enzyme activities over this time span. Gaps in the gut wall indicative of spent B-cells were observed in 17 of 39 copepods, with no trend over time or food treatment. Generally, only a single gap was seen in any one copepod. B-cells were vacuolated throughout the 24 h period. Morphometric analyses revealed a correlation between gut fluorescence and both maximum vacuole area and vacuole volume, as well as percent vacuolated cell volume, in the low-food treatment. The high-food treatment, which had a relatively smaller increase in gut fluorescence (1.5-fold) during the night cycle, showed no significant increase in vacuole size. B-cells appear to have a life-cycle greater than the diel feeding period, and while B-cell vacuoles respond to the ingested food during the diel cycle, production of B-cells does not appear to be a limiting factor in A. tonsa's feeding cycle.     

Photosynthesis and chlorophyll a fluorescence rhythms of marine phytoplankton

Circadian rhythms in photosynthesis were defined in field populations of phytoplankton. Measurements of carbon-dioxide fixation rates demonstrated that a diurnal periodicity of photosynthesis in samples incubated under natural light-dark (LD) cycles also were observed to continue in similar samples which had been photoadapted to constant dim light (LL) for 48 h. These changes in photosynthetic rates preceded sunset and sunrise, had daily amplitudes that ranged from 1.5 to 2.0, appeared to be independent of light-intensity, and displayed maxima about midday, while rates of dark fixation of carbon dioxide and the photosynthetic pigment content per cell were constant over the circadian cycle. Similar rhythmicity also was detected in room-temperature (22°C) chlorophyll a fluorescence yield, in both the obsence and presence of the photosynthesis inhibitor DCMU [3-(3,4-dichlorophenyl)-1, 1-dimethylurea]. However, the magnitude and timing of the fluorescence rhythm maxima seem to depend on wavelengths monitored and, in part, on the measuring technique used. Also, the circadian changes in the fluorescence intensity were abolished at low temperature (-60°C), and the shape of the emission spectra of chlorophyll fluorescence of cells in LD and LL did not change over time. The significance of the fluorescence rhythms with regard to chlorophyll a determinations and photosynthetic rates is discussed. It was concluded that there was sufficient similarity between circadian rhythms of photosynthesis in natural phytoplankton populations and in laboratory cultures of dinoflagellates to suggest that the mechanism of regulation may be the same for both of them.     

Effects of UV-B irradiation on growth and survival of Antarctic marine diatoms

Growth rate, survival, and stimulation of the production of UV-B (280 to 320 nm) absorbing compounds were investigated in cultures of five commonly occurring Antarctic marine diatoms exposed to a range of UV-B irradiances. Experimental UV-B exposures ranged from 20 to 650% of the measured peak surface irradiance at an Antarctic coastal site (0.533 J m^-2 s^-1). The five diatom species (Nitzschia lecointei, Proboscia alata, P. inermis, Thalassiosira tumida and Stellarima microtrias) appear capable of surviving two to four times this irradiance. In contrast to Phaeocystis cf. pouchetii, another major component of the Antarctic phytoplankton, the concentrations of pigments with discrete UV absorption peaks in diatoms were low and did not change significantly under increasing UV-B irradiance. Absorbance of UV-B by cells from which pigments had been extracted commonly greatly exceeded that of the pigments themselves. Most of this absorbance was due to oxidisable cell contents, with the frustule providing the remainder. Survival of diatoms did not correlate with absorption by either pigments, frustules or oxidisable cell contents, indicating that their survival under elevated UV-B irradiances results from processes other than screening mechanisms.     

Autecological investigation on marine diatoms 3. Thalassiosira nordenskioeldii and Chaetoceros diadema

The temperature range for the best competitive position by growth of Thalassiosira nordenskioeldii Cleve has been determined by comparing generation times. It ranges from-1.5° to 6°C. At these temperatures, especially at lower light intensities, it was one of the fastest growing species, whereas above 6°C many other species grew faster. High light intensities at increasing temperatures became damaging. A flowering of the cold oligo-eurytherm diatom T. nordenskioeldii occurs not only in the upper layers, but can also occur at greater depth simultaneously, because decreasing daylengths at 6°C had the least influence on its growth. Continuous light at 6°C had a positive influence on its growth. The start of the T. nordenskioeldii spring flowering under the Arctic Sea Ice is discussed in connection with the occurrence of enclosed marine diatoms in Polar Sea Ice. The influence of the winter temperature on the spring flowering of the North Sea, the southern border for flowerings of T. nordenskioeldii, is discussed. For Chaetoceros diadema (Ehrenberg) Gran the best competitive position by growth is reached at-1.5° to about 6°C. It has the best opportunity of reaching high cell numbers at the lowest temperatures of the range. The occurrence of the cold oligo-eurytherm diatom Ch. diadema in plankton samples at temperatures above 10°C need not be incorret, for the species did grow in cultures at 12° and 16°C. The wrong interpretation of the experimentally determined optimum temperature of e.g. T. nordenskioeldii caused a discrepancy between experimental results and field data that does not exist. The question is discussed whether ecologically it is relevant to talk about a temperature optimum. On account of the results of T. nordenskioeldii the question of the adaptation of diatom cultures for the start of the real experiments is discussed.     

Diel eclosion rhythm of a sublittoral population of the marine insect Pontomyia pacifica

Within the scope of studies on adaptation of insects to intertidal and sublittoral environments, correlations between the reproduction period of the short-lived chironomid Pontomyia pacifica Tokunaga and tidal conditions were examined at the only known Japanese location of this species in 1980. The larval habitat is situated, except for deeper tide pools in the lower midlittoral zone, mainly within the inner sublittoral zone, whose upper area is covered by Hypnea choroides, the dominating algae during the summer. The aerial adults emerge on the surface of the open sea and swarm there independent of the tidal situation above the submersed sublittoral habitat. Eclosion always started during dusk and all reproductive activities were ended within 2.5 h after sunset. The adaptation of P. pacifica (sublittoral habitat, diel eclosion after sunset with mass concentration of the adults on the water surface, no semilunar or lunar timing of the reproduction period and sinking egg masses) corresponds with those of the convergent marine chironomid Clunio balticus from Europe. On the basis of a few laboratory observations with P. pacifica and detailed experiments with C. balticus, it is supposed that the diel eclosion of P. pacifica is also controlled by an endogenous, circadian timing mechanism and the 24-h light-dark cycle as an environmental time cue.Communicated by O. Kinne, Oldendorf/Luhe     

Persistent diel rhythms in the chlorophyll fluorescence of marine phytoplankton species

The diel periodicities of in-vivo chlorophyll fluorescence and DCMU-enhanced chlorophyll fluorescence of 47 marine phytoplankton species were examined for 2 d in a 14 h L:10 h D light: dark cycle and then in continous light for another 2 to 3 d. Almost all phytoplankton species exhibit a more rapid increase in in-vivo fluorescence and DCMU-enhanced fluorescence during the light phase than during the dark phase. About one-half of the species examined exhibited persistent diel rhythms in continous light, indicating the operation of a biological clock. No phylogenetic or habitat related trends as to which species exhibited persistent rhythms were apparent. Of the phyla of eukaryotic phytoplankton adequately examined, none lacked biological clocks. Contrary to past hypotheses, some phytoplankton species maintain a persistent diel rhythm in a constant environment while reproducing at a rate greater than one division per day.     

Influence of light and temperature on the photosynthetic rate of marine benthic diatoms

The influence of light and temperature on photosynthetic rate as measured by C¹⁴O₂-fixation of marine benthic diatoms was investigated, using both intact sediment samples (Marshall et al., 1973) and suspensions of diatoms harvested by a lenstissue technique (Eaton and Moss, 1966). After C¹⁴-incubation, sediment samples were filtered, burned in a sample oxidizer, and their activity determined in a liquid scintillation counter. Photosynthetic rate of mixed field populations is saturated by a light intensity of approximately 10,000 lux; at still higher light intensities no photoinhibition was found. In contrast to the mixed field populations, unialgal cultures of the benthic diatom Amphiprora alata Kütz. exhibited strong photoinhibition at higher light intensities (10,000 to 60,000 lux). Within a range of 4° to 20°C, the photosynthetic rate increased about 10%/C^o. No differences in photosynthetic pattern were observed between epipelic and epipsammic species.     

Sinking rate response to depletion of nitrate,phosphate and silicate in four marine diatoms

The effect of nutrient (N, P, and Si) depletion on sinking rates was studied for two small (Skeletonema costatum (Grev.) Cleve and Chaetoceros gracile Schütt) and two large [Ditylum brightwellii (West) Grun and Coscinodiscus wailesii (Gran et Angst)] centric diatoms obtained from stock cultures. Each diatom was examined under conditions of (1) nutrient repletion (=log growth phase), (2) nutrient depletion (48 h without a given substrate), and (3) recovery (24h after addition of limiting substrate to nutrient-deplete populations). All nutrient-replete cultures displayed low sinking rates despite large differences in cell size. In nutrient-deplete populations, sinking rate was related to the kind of nutrient depleted and varied among species. Silicate depletion elicited by far the greatest increase in sinking rates in all 4 species, indicating that biochemical aspects of silicon metabolism are more important to buoyancy regulation than density-related variations in the amount of silicon per cell. Since N- and P-depletion caused lower sinking rates in 3 of the species, this observation calls for re-evaluation of the axiom that nutrient depletion necessarily causes increased sinking rates. The exception was Coscinodiscus wailesii, which sank faster under all types of nutrient limitation. In most cases, sinking rates typical of log-phase cultures were not regained within 24 h after the addition of limiting nutrient to nutrient-depleted populations. Ultimately, the length of the recovery period may be useful in identifying the metabolic processes responsible for buoyancy regulation in actively growing cells.     

Effect of temperature on nitrite excretion by three marine diatoms during nitrate uptake

Nitrite excretion during nitrate uptake by three nitrate-limited diatoms was measured at different temperatures. Phaeodactylum tricornutum Bohlin and Chaetoceros affinis Lauder Hustedt excreted nitrite over the whole range of physiological temperatures, whereas Thalassiosira pseudonana Hasle and Heimdal (Clone 13-1) excreted nitrite only at 25°C. Parallel to growth and nitrate-uptake rates, excretion rates increased exponentially with temperature, attaining a maximum between 20° and 25°C (optimum temperature range). At the end of nitrate uptake, when the nitrate concentration in the culture medium had decreased below 1 M, nitrite was reabsorbed at all temperatures, except when cells were in the dark or at very low light intensity. Nitrite uptake was also inhibited by the presence of nitrate in the medium. These results are discussed in relation to the formation and position of the maximum layer of primary nitrite in the thermocline, below the maximum layer of chlorophyll in stratified oceanic areas.     

Chlorophyll a fluorescence in marine centric diatoms: Responses of chloroplasts to light and nutrient stress

Observations at sea of large variations in the cellular fluorescence of phytoplankton prompted a study of the fluorescence responses in marine diatoms to light and nutrient stress. When older cultures of Lauderia borealis were exposed to intense light, the in vivo fluorescence of chlorophyll a declined within the first 2 min of exposure. This initial response to light stress appeared to be correlated with a contraction of the chloroplasts. Continued exposure led to a second decline in fluorescence, which required 30 to 60 min for completion. A movement of chloroplasts to the valvar ends of the cell caused this secondary response. Both the contraction and intracellular movement of chloroplasts appeared to be related to both photoinhibition of photosynthesis and diel fluctuations in cellular fluorescence. An investigation of continuous cultures of Cyclotella nana showed that in vivo chlorophyll a fluoresced more strongly in nitrogen-starved cells than in enriched ones. Photoinhibition of cellular fluorescence also increased with the cell's state of nitrogen deficiency.     

Interactions between two species of marine diatoms: Effects on their individual copper tolerance

The purpose of this study was to examine the effect of the interaction of two species of marine diatoms,Skeletonema costatum (Cleve) Greville andNitzschia thermalis (Ehrenberg) Auerswald, on their individual copper tolerances. The two species, obtained from stock cultures in 1989, were grown together at three copper concentrations (1, 4 and 5 × 10^–7 M added total copper). In the unialgal cultures that were used as controls, the two species grew as predicted from their tolerance tests. However, in mixed cultures,N. thermalis was the only species that exhibited growth, regardless of the copper concentration in the medium. Growth retardation ofS. costatum in the presence ofN. thermalis was attributed to an inhibitory exudate. The effect of the exudate appeared to have been temporary, as demonstrated by the extended lag phase and subsequent satisfactory exponential growth rate ofS. costatum. It is suggested that the exudate degraded within a period of 5 d (=lag phase) because exponential growth rate was resumed. It thus appears that the interaction between the two diatom species is more important in determining the survival ofS. costatum than its individual copper tolerance. This is not the case forN. thermalis. Such interactions would be unaccounted for in single-species toxicity tests. On the other hand, if they are known, prediction of how a community that includes these two species would respond to copper additions becomes possible.