Yellow water in La Jolla Bay,California, July 1980. I. A bloom of the dinoflagellate, Gymnodinium flavum Kofoid & Swezy

During the latter half of July, 1980, a bloom of Gymnodinium flavum Kofoid & Swezy caused water discoloration in La Jolla Bay, California. This naked dinoflagellate dominated the phytoplankton, numeri- cally and by volume, and was found in concentrations as high as 6.15 × 10³ cells · ml^?1. It was most abundant near the surface above a shallow (5–10 m), sharp thermocline and a nitracline at 10 m. Near the end of July, the depth of maximum phytoplankton abundance descended and water discoloration was no longer visible at the surface even though areal concentrations of G. flavum did not decrease. Concurrent with changes in the vertical distribution of the phytoplankton, warm, nutrient-depleted water moved into the area and nitrate availability in the upper 20 m of the water column was drastically reduced. Measure- ments of the chemical composition of the phytoplankton do not, however, indicate progressive nutrient stress during the period of environmental change. We conclude that shoaling of the thermocline and nitracline associated with apparent upwelling were conducive to development of the bloom and that advection of warmer water from offshore led to disappearance of yellow surface water from the bay.     

Hemolytically active components from P. parvum and G. breve toxins

Hemolytically active fractions were isolated from the toxins produced by the red-tide dinoflagellate $\text{Gymnodinium breve}$ (GBTX) and the chrysomonad $\text{Prymnesium parvum}$ (PPTX). High pressure liquid chromatography through bonded phase (ODS) silica columns using a gradient of methanol in chloroform yielded 6 major fractions from GBTX, 3 of which were hemolytic (HD₅₀=0.3?0.56 μg·ml^?1). None were ichthyotoxic. Of the 6 fractions obtained from PPTX, 4 were hemolytic (HD₅₀=0.013?2.8 μg·ml^?1) but only one (fraction 6) was ichthyotoxic. This fraction was ～ 2000 times more hemolytic than the crude PPTX (HD₅₀=33.2 μg·ml^?1). Analysis of their UV spectra indicates that the fractions within each group are closely related.     

Bimodal oxygen uptake in a freshwater air-breathing fish,Notopterus chitala

Measurements of bimodal oxygen uptake have been made in a freshwater air-breathing fish,Notopterus chitala at 29.0±1(S.D.)°C. xhe mean oxygen uptake from continuously flowing water without any access to air, was found to be 3.58±0.37 (S.E.) ml O₂ · h^?1 and 56.84+4.29 (S.E.) ml O₂ · kg^?1 · h^?1 for a fish weighing 66.92 + 11.27 (S.E.) g body weight. In still water with access to air, the mean oxygen uptake through the gills were recorded to be 2.49 ± 0.31 (S.E.) ml O₂ · h^?1 and 38.78 ± 1.92 (S.E.) ml O₂ · kg^?1 · h^?1 and through the accessory respiratory organs (swim-bladder) 6.04±0.87 (S.E.) ml O₂ · h^?1 and 92.32±2.91 (S.E.) ml O₂ · kg^?1 · h^?1 for a fish averaging 66.92±11.27 (S.E.) g. Out of the total oxygen uptake (131.10 ml O₂ · kg^?1 · h^?1), about 70% was obtained through the aerial route and the remainder 30% through the gills.     

PIGMENTS,FATTY ACIDS,AND STEROLS OF THE TOXIC DINOFLAGELLATE GYMNODINIUM CATENATUM1

Cultures and field samples of the toxic dinoflagellate Gymnodinium catenatum Graham from Tasmania, Australia, were analyzed for pigment, fatty acid, and sterol composition. Gymnodinium catenatum contained the characteristic pigments of photosynthetic dinoflagellates, including chlorophyll a, chlorophyll c₂, and the carotenoids peridinin, dinoxanthin, diadinoxanthin, diatoxanthin, and β,β-carotene. In midlogarithmic and early stationary phase cultures, the chlorophyll a content ranged 50–72 pg · cell^?1, total lipids 956–2084 pg · cell^?1, total fatty acids 426–804 pg · cell^?1, and total sterols 8–20 pg · cell^?1. The major fatty acids (in order of decreasing abundance) were 16:0, 22:6(n-3), and 20:5(n-3) (collectively 65–70% of the total fatty acids), followed by 16:1(n-7), 18:2(n-6), and 14:0. This distribution is characteristic of most dinoflagellates, except for the low abundance (<3%) of the fatty acid 18:5(n-3), considered by some authors to be a marker for dinoflagellates. The three major sterols were 4α-methyl-5α-cholest-7-en-3β-ol, 4α,23,24-trimethyl-5α-cholest-22E-en-3β-ol (the dinoflagellate sterol, dinosterol), and 4α,23,24-trimethyl-5α-cholest-7-en-3β-ol. These three sterols comprised about 75% of the total sterols in both logarithmic and early stationary phase cultures, and they were also found in high proportions (22–25%) in natural dinoflagellate bloom samples. 4-Desmethyl sterols, which are common in most microalgae, were only present in trace amounts in G. catenatum. The chemotaxonomic affinities of G. catenatum and the potential for using specific signature lipids for monitoring toxic dinoflagellate blooms are discussed.     

VEGETATIVE REPRODUCTION AND SEXUAL LIFE CYCLE OF THE TOXIC DINOFLAGELLATE GYMNODINIUM CATENATUM FROM TASMANIA,AUSTRALIA1

The toxic, chain-forming dinoflagellate Gymnodinium catenatum Graham was cultured from vegetative cells and benthic resting cysts isolated from estuarine waters in Tasmania, Australia. Rapidly dividing, log phase cultures formed long chains of up to 64 cells whereas stationary phase cultures were composed primarily of single cells (23-41 pm long, 27-36 pm wide). Vegetative growth (mean doubling time 3-4 days) was optimal at temperatures from 14.5-20° C, salinities of 23-34% and light irradiances of 50-300 μE·m^?2·s^?1. The sexual life cycle of G. catenatum was easily induced in a nutrient-deficient medium, provided compatible opposite mating types were combined (heterothallism). Gamete fusion produced a large (59-73 μm long, 50-59 μm wide) biconical, posteriorly biflagellate planozygote (double longitudinal flagellum) which after several days lost one longitudinal flagellum and gradually became subspherical in shape. This older planozygote stage persisted for up to two weeks before encysting into a round, brown resting cyst (42-52 μm diam; hypnozygote) with microreticulate surface ornamentation. Resting cysts germinated after a dormancy period as short as two weeks under our culture conditions, resulting in a single, posteriorly biflagellate germling cell (planomeiocyte). This divided to form a chain of two cells, which subsequently re-established a vegetative population. Implications for the bloom dynamics of this toxic dinoflagellate, a causative organism of paralytic shellfish poisoning, are discussed.     

THE PHYSIOLOGICAL ECOLOGY OF A FRESHWATER DINOFLAGELLATE BLOOM POPULATION: VERTICAL MIGRATION,NITROGEN LIMITATION,AND NUTRIENT UPTAKE KINETICS1,2

The motile freshwater dinoflagellate Gymnodinium bogoriense Klebs., which forms dense blooms in Jezre'el Valley water reservoirs (Israel) appears to be physiologically suited to exploit stratified environments, where it outcompetes all other phytoplankton types. The dense summer blooms (“red tides”) were found to be nitrogen-limited. The algae's competitive advantage, however, cannot result from superior uptake capabilities: its K_s (μmol NH₄·L^?1) for NH₄ was higher and its V_maxμmol NH₄·mg chlorophyll a^?1·h^?1) was lower than other phytoplankton types commonly occurring in the region. The competitive advantage of G. bogoriense probably stems from other physiological capabilities: dark ammonia and phosphorus assimilation and the ability to undertake diel vertical migration cycles between the upper photic water layers during the day and nutrient-rich deeper layers at night. These findings confirm the vertical nutrient retrieval hypothesis in migrating phytoplankton.     

CELL‐SPECIFIC EXTRACELLULAR PHOSPHATASE ACTIVITY OF DINOFLAGELLATE POPULATIONS IN ACIDIFIED MOUNTAIN LAKES1

High bulk extracellular phosphatase activity (PA) suggested severe phosphorus (P) deficiency in plankton of three acidified mountain lakes in the Bohemian Forest. Bioavailability of P substantially differed among the lakes due to differences in their P loading, as well as in concentrations of aluminum (Al) and its species, and was accompanied by species‐specific responses of phytoplankton. We combined the fluorescently labeled enzyme activity (FLEA) assay with image cytometry to measure cell‐specific PA in natural populations of three dinophyte species, occurring in all the lakes throughout May–September 2007. The mean cell‐specific PA varied among the lakes within one order of magnitude: 188–1,831 fmol · cell^?1 · h^?1 for Gymnodinium uberrimum (G. F. Allman) Kof. et Swezy, 21–150 fmol · cell^?1 · h^?1 for Gymnodinium sp., and 22–365 fmol · cell^?1 · h^?1 for Peridinium umbonatum F. Stein. To better compare cell‐specific PA among the species of different size, the values were normalized per unit of cell biovolume (amol · μm^?3 · h^?1) for further statistical analysis. A step‐forward selection identified concentrations of total and ionic Al together with pH as significant factors (P < 0.05, Monte Carlo permutation test), explaining cumulatively 57% of the total variability in cell‐specific PA. However, this cell‐specific PA showed an unexpected reverse trend compared to an overall gradient in P deficiency of the lake plankton. The autecological insight into dinophyte cell‐specific PA therefore suggested other factors, such as light availability, mixotrophy, and/or zooplankton grazing, causing further PA variations among the acidified lakes.     

Grazing impact on the cyanobacterium Microcystis aeruginosa by the heterotrophic flagellate Collodictyon triciliatum in an experimental pond

We estimated the grazing impact of the heterotrophic flagellate Collodictyon triciliatum on the harmful, bloom-forming cyanobacterium Microcystis aeruginosa in an experimental pond during a Microcystis bloom from summer to winter in 2010. For these experiments, we calculated the grazing rates from the digestion rate of C. triciliatum and its food vacuole contents. During the study period, M. aeruginosa exhibited one bloom event with a maximum density of 1.1 × 10⁵ cells ml^?1. The cell density of C. triciliatum fluctuated from below the detection limit to 291 cells ml^?1. The number of M. aeruginosa cells ingested by C. triciliatum food vacuoles ranged between 0.4 and 10.8 cells flagellate^?1, and the digestion rate of C. triciliatum at 25 °C was 0.73 % cell contents min^?1. The grazing rate of C. triciliatum on the M. aeruginosa prey was 0.2–6.9 cells flagellate^?1 h^?1, and its grazing impact was 0.0–25.3 % standing stock day^?1. The functional response of C. triciliatum to the M. aeruginosa prey followed the Michaelis–Menten model of significance (r ² = 0.873, p < 0.001) in our experimental systems, in which the prey concentration varied from 1.0 × 10⁴ to 2.1 × 10⁶ cells ml^?1. The maximum grazing rate was 6.2 prey cells grazer^?1 h^?1, and the half-saturation constant was 1.2 × 10⁵ cells ml^?1. We present evidence that C. triciliatum grazing explained the remarkable decrease in M. aeruginosa cell density in the pond. The present study is the first demonstration of the high potential of protistan grazing on M. aeruginosa to reduce cyanobacterial blooms.     

Strong small-scale temporal bacterial changes associated with the migrations of bloom-forming dinoflagellates

Bacterial abundances in nearshore Mediterranean planktonic environments tend to change seasonally by 10-fold. Strong daily changes in bacterial abundance, at least as large as seasonal range, occurred in the presence of large dinoflagellate populations performing daily vertical migrations. The daily variability of heterotrophic bacteria was associated with the daily migrations of a bloom of Alexandrium taylori in La Fosca Bay, and Gymnodinium impudicum in Barcelona harbor. Bacterial abundance in surface waters can change daily as much as from 1 × 10⁶ to 5 × 10⁶ with apparent net change rates of 0.24 h⁻¹. We suggest that the migrating dinoflagellates create microstructures exploited by the bacteria, and that the large algal populations (>10⁶ cells l⁻¹) make this microstructure visible with conventional sampling protocols. We also show evidence of the link between dinoflagellate abundance and relative bacterial activity in these waters, as measured by the percentage of bacteria with high nucleic acid content.     

PHYSIOLOGICAL VARIATION AND ELECTROPHORETIC BANDING PATTERNS OF GENETICALLY DIFFERENT SEASONAL POPULATIONS OF SKELETONEMA COSTATUM (BACILLARIOPHYCEAE)1

Clones of Skeletonema costatum (Grev.) Cl. isolated from Narragansett Bay, R.I., during different seasons were grouped according to their electrophoretic banding patterns. The growth rates, pg chlorophyll · cell^?1, carbon uptake · cell^?1· h^?1, and carbon uptake · pg chl^?1· h^?1 were measured at 20°C, in a 14:10 h L:D cycle at 180 μE · m^?2· s^?1. Statistically significant sources of variation were found among groups of clones in growth rate, pg chl · cell^?1, and carbon uptake · pg chl^?1· h^?1. It was concluded that there is a significant relationship between the physiological characteristics of clones isolated from populations in different seasons and patterns of genetic variation inferred from the electrophoretic studies. However, genetic diversity detected by banding patterns tends to underestimate the total genetic diversity in natural populations. The groups of clones most common in summer bloom populations had significantly higher growth rates, lower values of pg chl · cell^?1, and higher rates of carbon uptake · pg chl^?1· h^?1 at 20°C than did the group of clones most common in winter bloom populations. However, differences among groups in these parameters at 20°C alone cannot account for the seasonal cycling of genetically variable populations of Skeletonema in Narragansett Bay. The range of growth rates among clones of this species is 0.1–5.0 divisions · d^?1 under a single set of temperature and light conditions. Chlorophyll concentrations range from 0.2–1.7 pg chl · cell^?1 and carbon uptake · pg chl^?1· h^?1 varies by a factor of 7 among clones. The range of physiological variation in this species means that it is difficult to use laboratory studies of single clones to analyze the responses of natural populations of Skeletonema.     

SYMBIOSIS IN THE PLANKTONIC FORAMINIFER,ORBULINA UNIVERSA,AND THE ISOLATION OF ITS SYMBIOTIC DINOFLAGELLATE,GYMNODINIUM BÉII SP. NOV.1

The symbiotic association of the spinose planktonic foraminifer, Orbulina universa, with the dinoflagellate, Gymnodinium béii sp. nov., was examined with light and electron microscopy, and the symbiont was isolated into unialgal culture. The intact association is characterized by a diurnal movement of the symbionts from the distal regions of the spines during the day, to perialgal vacuoles within the host cytoplasm at night. This diurnal migration involves a daily endo- exocytotic cycle. Gymnodinium béii is non-motile and spindle-shaped within the host, whereas it is motile and gymnodinoid in shape when in culture. Ultrastructural examination revealed two or more stalked pyrenoids penetrated by lamellae, a typical dinokaryon nucleus and no trichocysts. A distinct ‘flange’projects over the sulcus from the hypocone. The swimming behavior of this dinoflagellate was characterized by intermittent darting events. Swimming speeds during a dart reached velocities of 770 μm. s^?1 as compared to a mean, non-darting swimming velocity of 126 μm. s^?1. Gymnodinium béii is eurythermal and division rates ranged between 0.16 and 0.65 divisions day^?1 for culture temperatures between 6.5 and 25° C respectively.     

The effect of food composition on ingestion,development, and survival of a harpacticoid copepod, Tisbe cucumariae Humes

Feeding experiments were carried out on the benthic harpacticoid copepod Tisbe cucumariae Humes, using seven different diets of various dried and ground macroalgae and marsh grass, algal Aufwuchs, diatoms, polychaete meat, and cereal. In short-term experiments (1 h), ¹⁴C-labelled foods were used to measure ingestion rate of non-ovigerous adult females (individual dry wt = 5.57 ± 2.49 μg). No significant difference was found among the rates at which all foods, except polychaete meat, were ingested (12.7 to 17.3 × 10^?2μg dry wt· ind^?1· h^?1). Polychaete meat was consumed faster (23.9 × 10^?2μg dry wt·ind ^?1· h^?1). The nutritional value of the foods was estimated in long-term experiments (22 days) by measuring development time and survival of T. cucumariae. Both these variables were significantly correlated with the nitrogen, protein content, and C:N ratio of the foods. No relation was found, however, with the amount of carbon, calories and available calories in the diets. Thus, nitrogen (protein) content of the food was the factor limiting secondary production of the copepods.     

Inorganic carbon uptake by phytoplankton in Vineyard Sound,Massachusetts. II. Comparative primary productivity and nutritional status of winter and summer assemblages

Using time-course, natural-light incubations, we assessed the rate of carbon uptake at a range of light intensities, the effect of supplemental additions of nitrogen (as NH₄⁺ or urea) on light and dark carbon uptake, and the rates of uptake of NH₄⁺ and urea by phytoplankton from Vineyard Sound, Massachusetts from February through August 1982. During the winter, photoinhibition was severe, becoming manifested shortly after the start of an incubation, whereas during the summer, there was little to no evidence of photoinhibition during the first several hours after the start of an incubation. At light levels which were neither photoinhibiting nor light limiting, rates of carbon uptake normalized per liter were high and approximately equal during winter and summer (22–23 μg C·l^?1 · h^?1), and low during spring (<10 μgC·l^?1· h^?1). In contrast, on a chlorophyll a basis, rates of carbon fixation were as high during spring (15–20μg C·μg Chl a^?1·h^?1), when concentrations of chlorophyll a were at the yearly minimum (<0.5 μg · l^?1) as during the summer, when chlorophyll a concentrations were substantially higher (0.8–1.3 μg · l^?1). Highest rates of NH₄⁺ and urea uptake were observed during summer, and at no time of the year was there evidence for severe nitrogen deficiency, although moderate nitrogen nutritional stress was apparent during the summer months.     

Utilisation de La méthode d'analyse par spectrométrie d'émission pour La détermination de 15N en écologie aquatique: Assimilation du nitrate par le phytoplancton en Mer de Banda (Indonésie)

A routine procedure for ¹⁵N determination with an optical emission spectrometer is described. The required amount of labelled sample (minimum 2 μg) is small and so it is possible to reduce the volume of the incubation-flasks (1 liter or less), which is advantageous when working on board ship. Samples are processed by the micro-Dumas method in sealed borosilicated discharge tubes. Combustion by products are trapped in liquid nitrogen. The method has been used successfully in the Banda Sea (cruise Corindon 4, N.O. Coriolis, in Indonesia) with labelled phytoplankton (¹⁴C and ¹⁵N). In surface waters specific uptake rates for N are in the range of 0.00005 · h^?1 to 0.0230 · h^?1 and of 0.0019 · h^?1 to 0.0295 · h^?1 for C.     

Contribution of Ciliated Microprotozoans and Dinoflagellates to the diet of three Copepod species in the Bay of Biscay

Predation of three calanoid copepods (Calanus helgolandicus, Temora longicornis and Centropages chierchiae) on phytoplankton, dinoflagellates and ciliates was estimated in the Gironde estuarine plume (Bay of Biscay) during winter by means of in situ incubations. Both phytoplankton and ciliates were part of the diet of those three species, while only Centropages chierchiae also included a significant portion of dinoflagellates in its diet. The clearance rates of Calanus helgolandicus for ciliates and phytoplankton reached 2.8 and 4.0 ml copepod^–1 h^–1, respectively, those of Temora longicornis were 3.2 and 1.8 ml copepod^–1 h^–1, and those of Centropages chierchiae were 4.3 and 0.8 ml copepod^–1 h^–1.Neither Calanus helgolandicus nor Temora longicornis selected dinoflagellates, given the low clearance rates measured for this prey category (0.05 and 0.03 ml copepod^–1 h^–1, respectively). By contrast, Centropages chierchiae included dinoflagellates in its diet, with a clearance rate of 4.9 ml copepod^–1 h^–1. Within a given prey category (ciliates or dinoflagellates), all three copepods selected larger prey types (>40 m) over smaller ones (40 m). This implies a better detection and capture of larger motile prey compared to small ones. The results are discussed with regard to the omnivorous feeding behavior of these copepods observed here, during a late winter phytoplankton bloom.     

Relationships between irradiance and photosynthesis for marine benthic green algae (Chlorophyta) of differing morphologies

Photosynthesis-irradiance relationships were determined in the field for five species of littoral and shallow sublittoral marine benthic green algae (Chlorophyta) of differing morphologies. Each species exhibited a linear increase in photosynthetic rate with increasing irradiance up to a maximum light-saturated value. Full sunlight (1405 to 1956 μE·m^?2·s^?1) inhibited photosynthesis of all species except the thick, optically dense, Codium fragile (Sur.) Har. Compensation irradiances ranged from 6.1 μE·m^?2·s^?1 for Enteromorpha intestinalis (L.) Link to 11.4 μE·m^?2·s^?1 for Ulva lobata (Kütz) S. & G. and did not reveal a consistent relationship to seaweed morphology. Saturation irradiances were determined statistically (I_k) and visually from graphical plots. with the latter technique resulting in values three to eight times higher and different comparative rankings of species than the former. I_k saturation irradiances were highest for Chaetomorpha linum (Müll.) Kütz. (81.9 μE·m^?2·s^?1) and lowest for Codium fragile (49.6 μE·m^?2·s^?1) and did not reveal a relationship with seaweed morphology. Regression equations describing light-limited photosynthetic rates and the relative magnitudes of the maximal net photosynthetic responses both strongly suggested a relationship with seaweed morphology. Highest net photosynthetic rates were obtained for the thin, sheet-like algae Ulva lobata (9.2 mg C·g dry wt^?1·h^?1), U. rigida C. Ag. (6.5 mg C·g dry wt^?1·h^?1) and the tubular form, Enteromorpha intestinalis (7.3 mg C·g dry wt^?1·h^?1), while lowest rates occurred for Codium fragile (0.9 mg C·g dry wt^?1·h^?1). Similarly, steepest light-limited slopes were found for the algae of simpler morphology, while the most gradual slope was determined for Codium fragile, the alga with greatest thallus complexity.     

THE TOXIC DINOFLAGELLATE GYMNODINIUM CATENATUM (DINOPHYCEAE) REQUIRES MARINE BACTERIA FOR GROWTH1

Interactions with the bacterial community are increasingly considered to have a significant influence on marine phytoplankton populations. Here we used a simplified dinoflagellate‐bacterium experimental culture model to conclusively demonstrate that the toxic dinoflagellate Gymnodinium catenatum H. W. Graham requires growth‐stimulatory marine bacteria for postgermination survival and growth, from the point of resting cyst germination through to vegetative growth at bloom concentrations (10³ cells · mL^?1). Cysts of G. catenatum were germinated and grown in unibacterial coculture with antibiotic‐resistant or antibiotic‐sensitive Marinobacter sp. DG879 or Brachybacterium sp., and with mixtures of these two bacteria. Addition of antibiotics to cultures grown with antibiotic‐sensitive strains of bacteria resulted in death of the dinoflagellate culture, whereas cultures grown with antibiotic‐resistant bacteria survived antibiotic addition and continued to grow beyond the 21 d experiment. Removal of either bacterial type from mixed‐bacterial dinoflagellate cultures (using an antibiotic) resulted in cessation of dinoflagellate growth until bacterial concentration recovered to preaddition concentrations, suggesting that the bacterial growth factors are used for dinoflagellate growth or are labile. Examination of published reports of axenic dinoflagellate culture indicate that a requirement for bacteria is not universal among dinoflagellates, but rather that species may vary in their relative reliance on, and relationship with, the bacterial community. The experimental model approach described here solves a number of inherent and logical problems plaguing studies of algal‐bacterium interactions and provides a flexible and tractable tool that can be extended to examine bacterial interactions with other phytoplankton species.     

Phytoplankton ecology in the Southern Benguela current. III. Dynamics of a bloom

The development of a phytoplankton bloom was studied by placing a drogue in a patch of cold upwelled water and following the water mass for 4–5 days. Chaetoceros compressus Laud and Skeletonemacostatum (Grev.) Cleve dominated the bloom which reached its peak in 3 days. In this period chlorophyll a concentrations increased by 19.2 mg · m^?3 in the euphotic zone while the concomitant decrease in nitrate concentration was 18.7 mg-at. NO₃-N · m^?3. There was an overall increase in the concentration of protein with the highest concentration (412.9mg · m^?3) being measured just prior to the peak of the bloom. Carbohydrate concentrations increased rapidly during the day but decreased at night. The pattern of carbon-14 assimilation at the 50% light intensity was characterised by high activity in the polysaccharide fraction as the bloom developed, but at the peak of the bloom a greater percentage of the label was found in the ethanol-soluble fraction. The percent incorporation into protein was greater at night than during the day. These physiological changes are related to the growth pattern of the bloom.