Growth of dinoflagellates, Ceratium furca and Ceratium fusus in Sagami Bay, Japan: The role of vertical migration and cell division

To better understand the mechanism underlying the bloom outbreaks of dinoflagellates, Ceratium furca, and Ceratium fusus in the temperate coastal area of Sagami Bay, we investigated the diel changes of vertical migration, swimming speed, cell volume, and cell division. Our results from both the field and laboratory indicate that C. furca and C. fusus can migrate vertically between surface and sub-surface layers to avoid strong sunlight (>1000 μmol m⁻² s⁻¹). Diel vertical migration (DVM) of C. furca was observed in the laboratory, while that of C. fusus was not observed. C. furca demonstrated a constant DVM rhythm, i.e., their cells began to descend from the surface before the light was extinguished, and ascended into the surface before the light was turned on. The downward and upward migrations of the cells occurred at every 3 h before turning on and off the light, suggesting that the DVM pattern was independent of nutrient concentration. The swimming speeds of C. furca (avg. 250 μm s⁻¹) were always faster than those of C. fusus (avg. 75 μm s⁻¹). In addition, the speeds of C. furca during light periods were faster than those during dark periods, whereas the speeds of C. fusus remained relatively constant. A higher proportion of dividing cells was recorded near dawn (05:00–07:00 h). Cell volumes of C. furca and C. fusus did not markedly change between 12:00 and 21:00 h, but gradually increased until 03:00 h and then sharply decreased. Furthermore, the cell volume of the two Ceratium species was significantly shifted to the temporal pattern of cell division. Combined with the DVM manner of two Ceratium and cell division timing, only C. furca divided at the bottom, and then moved toward the surface shortly before the dark to light transition. Based on our observations, C. furca has an ecological advantage due to their DVM activity, since nutrients can be obtained well in the near bottom layers, while during the daytime, light present in nutrient-depleted surface water can be obtained using their high swimming speed. On the other hand, C. fusus stimulated by low salinity conditions, might be dependent on external environmental conditions such as additional nutrients following freshwater discharge by heavy rainfall because they may not perform active DVM due to a slow swimming ability. Our findings support that specific characteristics, including the DVM behavior in C. furca, yield a competitive advantage over C. fusus in Sagami Bay.     

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.     

Growth of dinoflagellates, Ceratium furca and Ceratium fusus in Sagami Bay, Japan: The role of nutrients

In order to study the influence of nutrients on the growth characteristics of the dominant dinoflagellates, Ceratium furca and Ceratium fusus, in the temperate coastal area of Sagami Bay, Japan, we conducted field monitoring from January 2000 to December 2005 and performed laboratory culture experiments. In the field study, population densities of C. furca and C. fusus were high, even in low nutrient concentrations (N: 1.58 μM, P: 0.17 μM). Both species were more abundant in the surface and sub-surface layers than in the bottom layers during the stratification periods. In the laboratory study, the specific growth rates of C. furca and C. fusus increased gradually along with increasing nutrients up to the T₅ (N: 5 μM, P: 0.5 μM) and T₁₀ (N: 10 μM, P: 1 μM) concentration levels, after which the growth rate plateaued at the T₅₀ (N: 50 μM, P: 5 μM) concentration level. In contrast, the nutrient uptake rates of both species continuously increased, indicating “luxury consumption”, i.e., excessive cellular storage not related to growth rate. The half-saturation constants (K_s) of C. furca for nitrate (0.49 μM) and phosphate (0.05 μM) were slightly higher than C. fusus (0.32 and 0.03 μM, respectively). We offer two reasons why the two Ceratium population densities were maintained at high levels in low nutrient conditions. First, these two species have a competitive advantage over other algal species because of low K_s values and specific characteristics for nutrient uptake such as luxury consumption. Their ability to obtain nutrients through alternative methods, such as phagotrophy, might contribute to bloom formation and population persistence. Second, the cell densities of both Ceratium species increased along with nitrate concentrations in the media even when phosphorus was held constant. In particular, the growth of C. furca was directly supported by various nitrogen sources such as nitrate, ammonium, and urea, although the highest growth rates were observed only in the nitrate-enriched cultures. Our field and laboratory results revealed that the growth rates of the two Ceratium species increased readily in high N:P nutrient conditions (i.e., conditions of P limitation) indicating an advantage over other algal species in phosphorus-limited environments such as Sagami Bay.     

Killing potential protist predators as a survival strategy of the newly described dinoflagellate Alexandrium pohangense

Blooms caused by some species belonging to the dinoflagellate genus Alexandrium are known to cause large-scale mortality of fish. Thus, the dynamics of these species is important and of concern to scientists, officials, and people in the aquaculture industry. To understand the dynamics of such species, their growth and mortality due to predation need to be assessed. The newly described dinoflagellate Alexandrium pohangense is known to grow slowly, with a maximum autotrophic growth rate of 0.1 d⁻¹. Thus, it may not form bloom patches if its mortality due to predation is high. Therefore, to explore the mortality of A. pohangense due to predation, feeding on this species by the common heterotrophic dinoflagellates Gyrodinium dominans, Gyrodinium moestrupii, Luciella masanensis, Noctiluca scintillans, Oxyrrhis marina, Oblea rotunda, Polykrikos kofoidii, and Pfiesteria piscicida, as well as by the ciliate Tiarina fusus, was examined. None of these potential predators was able to feed on A. pohangense. In contrast, these potential predators were killed and their bodies were dissolved when incubated with A. pohangense cells or cell-free culture filtrates. The survival of G. moestrupii, O. marina, P. kofoidii, and T. fusus on incubation with 10 cells ml⁻¹ of A. pohangense was 20–60%, while that at the equivalent culture filtrates was 20–70%. With increasing A. pohangense cell-concentration (up to 1000 cells ml⁻¹ or equivalent culture filtrates), the survival rate of G. moestrupii, O. marina, P. kofoidii, and T. fusus rapidly decreased. The lethal concentration (LC₅₀) for G. moestrupii, O. marina, P. kofoidii, and T. fusus at the elapsed time of 24 h with A. pohangense cells (cultures of 11.4, 13.3, 1.6, and 3.3 cells ml⁻¹, respectively) was lower than that with A. pohangense filtrates (culture filtrates of 35.5, 30.6, 5.5, and 5.0 cells ml⁻¹, respectively). Furthermore, most of the ciliates and heterotrophic dinoflagellates in the water collected from the coast of Tongyoung, Korea, were killed when incubated with cultures of 1000 A. pohangense cells ml⁻¹ and equivalent culture filtrates. The relatively slow growing A. pohangense may form blooms by reducing mortality due to predation through killing potential protist predators.     

Growth of eight Gambierdiscus (Dinophyceae) species: Effects of temperature,salinity and irradiance

Little is known about how the growth of individual Gambierdiscus species responds to environmental factors. This study examined the effects of temperature (15–34 °C), salinity (15–41) and irradiance (2–664 μmol photons m⁻² s⁻¹) on growth of Gambierdiscus: G. australes, G. belizeanus, G. caribaeus, G. carolinianus, G. carpenteri, G. pacificus and G. ruetzleri and one putative new species, Gambierdiscus ribotype 2. Depending on species, temperatures where maximum growth occurred varied between 26.5 and 31.1 °C. The upper and lower thermal limits for all species were between 31–34 °C and 15–21 °C, respectively. The shapes of the temperature vs. growth curves indicated that even small differences of 1–2 °C notably affected growth potentials. Salinities where maximum growth occurred varied between 24.7 and 35, while the lowest salinities supporting growth ranged from <14 to 20.9. These data indicated that Gambierdiscus species are more tolerant of lower salinities than is generally appreciated. Growth of all species began to decline markedly as salinities exceed 35.1–39.4. The highest salinity tested in this study (41), however, was lethal to only one species, Gambierdiscus ribotype 2. The combined salinity data indicated that differences in salinity regimes may affect relative species abundances and distributions, particularly when salinities are <20 and >35. All eight Gambierdiscus species were adapted to relatively low light conditions, exhibiting growth maxima at 50–230 μmol photons m⁻² s⁻¹ and requiring only 6–17 μmol photons m⁻² s⁻¹ to maintain growth. These low light requirements indicate that Gambierdiscus growth can occur up to 150 m depth in tropical waters, with optimal light regimes often extending to 75 m. The combined temperature, salinity and light requirements of Gambierdiscus can be used to define latitudinal ranges and species-specific habitats, as well as to inform predictive models.     

Lethal effects of ichthyotoxic raphidophytes,Chattonella marina,C. antiqua,and Heterosigma akashiwo,on post-embryonic stages of the Japanese pearl oyster,Pinctada fucata martensii

The inimical effects of the ichthyotoxic harmful algal bloom (HAB)-forming raphidophytes Heterosigma akashiwo, Chattonella marina, and Chattonella antiqua on the early-life stages of the Japanese pearl oyster Pinctada fucata martensii were studied. Fertilized eggs and developing embryos were not affected following exposure to the harmful raphidophytes; however, all three algal species severely affected trochophores and D-larvae, early-stage D-larvae, and late-stage pre-settling larvae. Exposure to C. marina (5 × 10² cells ml⁻¹), C. antiqua (10³ cells ml⁻¹), and H. akashiwo (5 × 10³ cells ml⁻¹) resulted in decreased success of metamorphosis to the trochophore stage. A complete inhibition of trochophore metamorphosis was observed following exposure to C. antiqua at 5 × 10³ cells ml⁻¹ and C. marina at 8 × 10³ cells ml⁻¹. In all experiments, more than 80% of newly formed trochophores were anomalous, and in the case of exposure to H. akashiwo at 10⁵ cells ml⁻¹ more than 70% of D-larvae were anomalous. The activity rates of D-larvae (1-day-old) were significantly reduced following exposure to C. antiqua (8 × 10³ cells ml⁻¹, 24 h), C. marina (8 × 10³ cells ml⁻¹, 24 h), and H. akashiwo (10⁴ cells ml⁻¹, 24 h). The activity rates of pre-settling larvae (21-day-old) were also significantly reduced following exposure to C. antiqua (10³ cells ml⁻¹, 24 h), C. marina (8 × 10³ cells ml⁻¹, 24 h), and H. akashiwo (5 × 10⁴ cells ml⁻¹, 24 h). Significant mortalities of both larval stages were induced by all three raphidophytes, with higher mortality rates registered for pre-settling larvae than D-larvae, especially following exposure to C. marina (5 × 10²–8 × 10³ cells ml⁻¹, 48–86 h) and C. antiqua (10³–8 × 10³ cells ml⁻¹, 72–86 h). Contact between raphidophyte cells and newly metamorphosed trochophores and D-larvae, 1-day-old D-larvae, and 21-day-old larvae resulted in microscopic changes in the raphidophytes, and then, in the motile early-life stages of pearl oysters. Upon contact and physical disturbance of their cells by larval cilia, H. akashiwo, C. marina and C. antiqua became immotile and shed their glycocalyx. The trochophores and larvae were observed trapped in a conglomerate of glycocalyx and mucus, most probably a mixture of larval mucous and raphidophyte tricosyts and mucocytes. All motile stages of pearl oyster larvae showed a typical escape behavior translating into increased swimming in an effort to release themselves from the sticky mucous traps. The larvae subsequently became exhausted, entrapped in more heavy mucous, lost their larval cilia, sank, become immotile, and died. Although other toxic mediators could have been involved, the results of the present study indicate that all three raphidophytes were harmful only for motile stages of pearl oysters, and that the physical disturbance of their cells upon contact with the ciliary structures of pearl oyster larvae initiated the harmful mechanism. The present study is the first report of lethal effects of harmful Chattonella spp. towards larvae of a bivalve mollusc. Blooms of H. akashiwo, C. antiqua and C. marina occur in all major cultivation areas of P. fucata martensii during the developmental period of their larvae. Therefore, exposure of the motile early-life stages of Japanese pearl oysters could adversely affect their population recruitment. In addition, the present study shows that further research with early-life development of pearl oysters and other bivalves could contribute to improving the understanding of the controversial harmful mechanisms of raphidophytes in marine organisms.     

Thresholds for morphological response to light reduction for four tropical seagrass species

Seagrasses worldwide are highly vulnerable to, and at increasing risk from reduced light availability, and robust light thresholds are required for evaluating future impacts of changing light conditions. We tested the morphological response (shoot density and growth) of four Indo-West Pacific seagrass species (Cymodocea serrulata, Halodule uninervis, Halophila ovalis and Zostera muelleri) to six daily light levels ranging from 0 to 23 mol m⁻² d⁻¹ (0–70% surface irradiance) in cool (∼23 °C) and warm temperatures (∼28 °C) over 14 weeks. The impact of light limitation on shoot densities and growth rates was higher at warm than at cool temperatures, and for Z. muelleri and H. ovalis than for C. serrulata and H. uninervis, in terms of both the time taken for the low light treatment to take effect and the predicted time to shoot loss (e.g. 17–143 days at 0 mol m⁻² d⁻¹). Using fitted curves we estimated temperature-dependent thresholds (with estimates of uncertainty) for 50% and 80% protection of growth and shoot density, defined here as “potential light thresholds” in recognition that they were derived under experimental conditions. Potential light thresholds that maintained 50% and 80% of seagrass shoot density fell within the ranges 1.1–5.7 mol m⁻² d⁻¹ and 3.8–10.4 mol m⁻² d⁻¹, respectively, depending on temperature and species. Light thresholds calculated in separate in situ studies for two of the same species produced comparable results. We propose that the upper (rounded) values of 6 mol m⁻² d⁻¹ and 10 mol m⁻² d⁻¹ can be used as potential light thresholds for protecting 50% and 80% of shoot density for these four species over 14 weeks. As management guidelines should always be more conservative than thresholds for biological declines, we used error estimates to provide a quantitative method for converting potential light thresholds into guidelines that satisfy this criterion. The present study demonstrates a new approach to deriving potential light thresholds for acute impacts, describes how they can be applied in management guidelines and quantifies the timescales of seagrass decline in response to light limitation. This method can be used to further quantify cumulative impacts on potential light thresholds.     

Mixotrophy in the newly described dinoflagellate Yihiella yeosuensis: A small,fast dinoflagellate predator that grows mixotrophically,but not autotrophically

To investigate tropical roles of the newly described Yihiella yeosuensis (ca. 8 μm in cell size), one of the smallest phototrophic dinoflagellates in marine ecosystems, its trophic mode and the types of prey species that Y. yeosuensis can feed upon were explored. Growth and ingestion rates of Y. yeosuensis on its optimal prey, Pyramimonas sp. (Prasinophyceae), as a function of prey concentration were measured. Additionally, growth and ingestion rates of Y. yeosuensis on the other edible prey, Teleaulax sp. (Cryptophyceae), were also determined for a single prey concentration at which both these rates of Y. yeosuensis on Pyramimonas sp. were saturated. Among bacteria and diverse algal prey tested, Y. yeosuensis fed only on small Pyramimonas sp. and Teleaulax sp. (both cell sizes = 5.6 μm). With increasing mean prey concentrations, both specific growth and ingestion rates of Y. yeosuensis increased rapidly before saturating at a mean Pyramimonas concentration of 109 ng C mL⁻¹ (2725 cells mL⁻¹). The maximum growth rate (mixotrophic growth) of Y. yeosuensis fed with Pyramimonas sp. at 20 °C under a 14:10-h light-dark cycle of 20 μE m⁻² s⁻¹ was 1.32 d⁻¹, whereas the growth rate of Y. yeosuensis without added prey was 0.026 d⁻¹. The maximum ingestion rate of Y. yeosuensis fed with Pyramimonas sp. was 0.37 ng C predator⁻¹ d⁻¹ (9.3 cells predator⁻¹ d⁻¹). At a Teleaulax concentration of 1130 ng C mL⁻¹ (66,240 cells mL⁻¹), growth and ingestion rates of Y. yeosuensis fed with Teleaulax sp. were 1.285 d⁻¹ and 0.38 ng C predator⁻¹ d⁻¹ (22.4 cells predator⁻¹ d⁻¹), respectively. Thus, Y. yeosuensis rarely grows without mixotrophy, and mixotrophy supports high growth rates in Y. yeosuensis. Y. yeosuensis has the highest maximum mixotrophic growth rate with the exception of Ansanella graniferaamong engulfment feeding mixotrophic dinoflagellates. However, the high swimming speed of Y. yeosuensis (1572 μm s⁻¹), almost the highest among phototrophic dinoflagellates, may prevent autotrophic growth. This evidence suggests that Y. yeosuensis may be an effective mixotrophic dinoflagellate predator on Pyramimonas and Teleaulax, and occurs abundantly during or after blooms of these two prey species.     

Locomotion at –1.0°C: burst swimming performance of five species of Antarctic fish

We investigated the burst swimming performance of five species of Antarctic fish at −1.0°C. The species studied belonged to the suborder, Notothenioidei, and from the families, Nototheniidae and Bathydraconidae. Swimming performance of the fish was assessed over the initial 300 ms of a startle response using surgically attached miniature accelerometers. Escape responses in all fish consisted of a C-type fast start; consisting of an initial pronounced bending of the body into a C-shape, followed by one or more complete tail-beats and an un-powered glide. We found significant differences in the swimming performance of the five species of fish examined, with average maximum swimming velocities (U_max) ranging from 0.91 to 1.39 m s⁻¹ and maximum accelerations (A_max) ranging from 10.6 to 15.6 m s⁻². The cryopelagic species, Pagothenia borchgrevinki, produced the fastest escape response, reaching a U_max and A_max of 1.39 m s⁻¹ and 15.6 m s⁻², respectively. We also compared the body shapes of each fish species with their measures of maximum burst performance. The dragonfish, Gymnodraco acuticeps, from the family Bathdraconidae, did not conform to the pattern observed for the other four fish species belonging to the family Nototheniidae. However, we found a negative relationship between buoyancy of the fish species and burst swimming performance.     

Seasonal variations in photosynthetic irradiance response curves of macrophytes from a Mediterranean coastal lagoon

The main photosynthesis and respiration parameters (dark respiration rate, light saturated production rate, saturation irradiance, photosynthetic efficiency) were measured on a total of 23 macrophytes of the Thau lagoon (2 Phanerogams, 5 Chlorophyceae, 10 Rhodophyceae and 6 Phaeophyceae). Those measurements were performed in vitro under controlled conditions, close to the natural ones, and at several seasons. Concomitantly, measurements of pigment concentrations, carbon, phosphorous and nitrogen contents in tissues were performed. Seasonal intra-specific variability of photosynthetic parameters was found very high, enlightening an important acclimatation capacity. The highest photosynthetic capacities were found for Chlorophyceae (e.g. Monostroma obscurum thalli at 17 °C, 982 μmol O₂ g⁻¹ dw h⁻¹ and 9.1 μmol O₂ g⁻¹ dw h⁻¹/μmol photons m⁻² s⁻¹, respectively for light saturated net production rate and photosynthetic efficiency) and Phanerogams (e.g. Nanozostera noltii leaves at 25 °C, 583 μmol O₂ g⁻¹ dw h⁻¹ and 2.6 μmol O₂ g⁻¹ dw h⁻¹/μmol photons m⁻² s⁻¹ respectively for light saturated net production rate and photosynthetic efficiency). As expected, species with a high surface/volume ratio were found to be more productive than coarsely branched thalli and thick blades shaped species. Contrary to R_d (ranging 6.7–794 μmol O₂ g⁻¹ dw h⁻¹, respectively for Rytiphlaea tinctoria at 7 °C and for Dasya sessilis at 25 °C) for which a positive relationship with water temperature was found whatever the species studied, the evolution of P/I curves with temperature exhibited different responses amongst the species. The results allowed to show summer nitrogen limitation for some species (Gracilaria bursa-pastoris and Ulva spp.) and to propose temperature preferences based on the photosynthetic parameters for some others (N. noltii, Zostera marina, Chaetomorpha linum).     

Factors controlling Zostera marina L. growth in the eastern and western Pacific Ocean: Comparisons between Korea and Oregon,USA

Zostera marina distribution is circum-global and tolerates a wide range of environmental conditions. Consequently, it is likely that populations have adapted to local environmental conditions of light, temperature and nutrient supply. We compared Z. marina growth dynamics over a 2-year period in relation to environmental characters at Jindong Bay, South Korea and Yaquina Bay, Oregon, USA. Water temperature in Jindong Bay showed stronger seasonal variation (summer–winter ΔT = 20 °C) than in Yaquina Bay (summer–winter ΔT < 5 °C). Underwater irradiance in Jindong Bay exhibited a winter maximum, while in Yaquina Bay underwater light exhibited a summer maximum. Integrated annual underwater irradiance during 2003 was 2200 and 1200 mol photons m⁻² year⁻¹ in Korea and Oregon, respectively. Z. marina shoot density, biomass and integrated production were not significantly different between the two study sites. Seasonal Z. marina growth in Jindong Bay appeared to be controlled by temperature and light, while the growth pattern in Yaquina Bay suggested light regulation. Several seagrass parameters were correlated to phosphate concentrations, even though nutrients did not appear limiting. Despite differences in environmental factors, relative growth rates and temporal growth dynamics between study sites, integrated annual leaf production was quite similar at 335 and 353 g DW m⁻² year⁻¹ in the Jindong and Yaquina Bay study sites. We suggest that Z. marina net productivity is acclimated to the local environmental conditions and may be a general characteristic of temperate seagrass populations.     

Evaluation of the effects of light intensity on growth of the benthic dinoflagellate Ostreopsis sp. 1 using a newly developed photoirradiation-culture system and a novel regression analytical method

Benthic dinoflagellates of the genus Ostreopsis are found all over the world in temperate, subtropical, and tropical coastal regions. Our recent studies revealed that a putative “cryptic” species of Ostreopsis ovata is present widely along Japanese coasts. This organism, Ostreopsis sp. 1, possesses palytoxin analogs and thus its toxic blooms may be responsible for potential toxification of marine organisms. To evaluate the bloom dynamics of Ostreopsis sp. 1, the present study examined the growth responses of Ostreopsis sp. 1 strain s0716 to various light intensities (photon flux densities: μmol photons m⁻² s⁻¹) using a newly devised photoirradiation-culture system. This novel system has white light-emitting diodes (LEDs) capable of more closely simulating the wavelength spectrum of light entering the oceanic water column than do fluorescent tubes and halogen lamps. In this system, the light intensity of the white LEDs was reduced through two polarizing filters by varying the rotation angles of the filters. Thereby, the new system was capable of culturing microalgae under well-controlled light intensity conditions. Ostreopsis sp. 1 grew proportionally when light intensity was increased from 49.5 to 199 μmol photons m⁻² s⁻¹, but its growth appeared to be inhibited slightly at ≥263 μmol photons m⁻² s⁻¹. The relationship between observed growth rates and light intensity was calculated at R > 0.99 (P < 0.01) using a regression analysis with a modified equation of the photosynthesis-light intensity (P-L) model. The equation determined the critical light intensities for growth of Ostreopsis sp. 1 and the organism's growth potential as follows: (1) the threshold light intensity for growth: 29.8 μmol photons m⁻² s⁻¹; (2) the optimum light intensity (L_m) giving the maximum growth rate (μ_max = 0.659 divisions day⁻¹): 196 μmol photons m⁻² s⁻¹; (3) the optimum light intensity range (L_opt) giving ≥95% μ_max: 130–330 μmol photons m⁻² s⁻¹; (4) the semi-optimum range (L_sopt) giving ≥80% μ_max: 90 to over 460 μmol photons m⁻² s⁻¹. The L_sopt represents 4.5–23% ambient light intensity present in surface waters off of a temperate region of the Japanese coast, Tosa Bay; putatively, this semi-optimum range of light intensity appears at depth of 12.9–27.8 m. Considering these issues, our data indicate that Ostreopsis sp. 1 in coastal environments may form blooms at ca. ∼28 m depth in regions along Japanese coasts.     

Developing a methodology for estimating the drag in front-crawl swimming at various velocities

We aimed to develop a new method for evaluating the drag in front-crawl swimming at various velocities and at full stroke. In this study, we introduce the basic principle and apparatus for the new method, which estimates the drag in swimming using measured values of residual thrust (MRT). Furthermore, we applied the MRT to evaluate the active drag (Da) and compared it with the passive drag (Dp) measured for the same swimmers. Da was estimated in five-stages for velocities ranging from 1.0 to 1.4 m s⁻¹; Dp was measured at flow velocities ranging from 0.9 to 1.5 m s⁻¹ at intervals of 0.1 m s⁻¹. The variability in the values of Da at MRT was also investigated for two swimmers. According to the results, Da (Da = 32.3 v^3.3, N = 30, R² = 0.90) was larger than Dp (Dp = 23.5 v^2.0, N = 42, R² = 0.89) and the variability in Da for the two swimmers was 6.5% and 3.0%. MRT can be used to evaluate Da at various velocities and is special in that it can be applied to various swimming styles. Therefore, the evaluation of drag in swimming using MRT is expected to play a role in establishing the fundamental data for swimming.     

Uptake and allocation of 13C by Enhalus acoroides at sites differing in light availability

Carbon fixation and allocation were studied using ¹³C incubation and leaf marking techniques in mature monospecific stands of Enhalus acoroides L.f. Royle in August 1998 and January 1999 in Banten Bay, Indonesia. The highest rate of ¹³C uptake (>0.008 g ¹³C g C⁻¹ d⁻¹) was found in the middle to distal parts of leaves of E. acoroides. Young and senescing leaves numbers had lower ¹³C incorporation compared to mature leaves. The incorporation of ¹³C by epiphytes on the leaves was higher than that of the leaves themselves (>0.01 g ¹³C g C⁻¹ d⁻¹). The results showed that turbidity of the water influenced the leaf growth, productivity and Relative Growth Rate of E. acoroides, which were lower at Kepuh Island, the more turbid site. However, at Kepuh Island, where the water column was turbid, the plant could still harvest sufficient light for an uptake rate of ¹³C, higher than the uptake rates at Kubur and Panjang Islands, stations with a much more transparent water column (on average 0.0047 g ¹³C g C⁻¹ d⁻¹ at Kepuh Island, versus 0.0045 g ¹³C g C⁻¹ d⁻¹ at Panjang Island and 0.0034 g ¹³C g C⁻¹ d⁻¹ at Kubur Island). There was evidence that ¹³C was exported from the incubated shoots to the roots and rhizomes and to neighboring shoots of E. acoroides in clear water, but not in turbid water. We suggest that single shoots of E. acoroides are able to grow in turbid water under low light conditions. They assimilate sufficient carbon for their own maintenance but are not able to export to neighboring plant parts.     

Sex-specific light acclimation of Chara canescens (Charophyta)

Bisexual populations of the charophyte Chara canescens (Desv. et Loisel. in Loisel., 1810) containing male and female individuals are rarely found. Two experiments were carried out to study whether male and female algae from the same site exhibit different physiological capacities, especially with respect to light acclimation.Algae from two different shore levels and from laboratory cultures acclimated to six irradiance conditions (35–500 μmol photons m⁻² s⁻¹) were compared. Field measurements showed that both female and male algae of C. canescens are able to acclimate to daily changes in solar irradiance. The quantum yield of Photosystem II (PSII) decreased with increasing irradiance in the morning and increased with decreasing irradiance in the afternoon. Growth experiments showed increasing growth rates from 35 μmol photons m⁻² s⁻¹ (∼7 mg FW) up to 500 μmol photons m⁻² s⁻¹ (∼27 mg FW) in female and male C. canescens. The irradiance saturation point for photosynthesis (E_k) was about 140 μmol m⁻² s⁻¹ for both sexes within the whole range of acclimation irradiances. The maximum photosynthesis rate at saturating irradiances (P_max) of male algae was highest at E_k, whereas P_max of female algae was highest at 500 μmol photons m⁻² s⁻¹. The photosynthetic efficiency in the light-limited range (α) increased in female C. canescens and decreased in male C. canescens. The ratio of the non-photochemical quenching parameter (NPQ) to the relative electron transport rates rETR_(MT) increased in both sexes with irradiance, but showed a steeper increase in male than in female algae. Pigment analysis showed similar acclimation pattern for male and female C. canescens. Chl a/Chl b ratios of both sexes were constant over the whole range of E_g, whereas Chl a/carotenoid ratios in male and female C. canescens decreased from 70 μmol photons m⁻² s⁻¹ upwards. Pigment analysis pointed out that the carotenes α-, β- and γ-carotene were more prominent in male than in female algae.Our results indicate that female C. canescens are more efficient in light acclimation than male algae from the same site. Nevertheless, further investigations of bisexual C. canescens populations resolving CO₂-uptake mechanisms and/or genetic differences are needed.     

Growth and domoic acid content of Pseudo-nitzschia australis isolated from northwestern Baja California,Mexico, cultured under batch conditions at different temperatures and two Si:NO3 ratios

Domoic acid (DA) poisoning in the southern part of the California Current System has been associated typically with blooms of Pseudo-nitzschia australis. The environmental variables that promote growth and DA production in the Mexican part of this system have not been identified. The present study investigated the effect of temperature and two nutrient ratios on the growth characteristics and DA content of two (BTS-1, BTS-2) P. australis strains isolated from the Pacific coast of northern Baja California peninsula, México. Of the different temperatures assayed (10, 12, 14, 15, 18 and 20 °C), the maximum cell abundance was detected at 12 °C for BTS-2 and 14 °C for BTS-1. The highest maximum specific growth rate (1.69 day⁻¹) was measured at 15 °C for BTS-2. With the exception of cells maintained at 15 °C, growth characteristics were similar in P. australis cultured in a high Si:NO₃ (2.5) or low Si:NO₃ (0.5) ratio at each temperature. Dissolved (dDA) and cellular (cDA) DA content measured at the stationary phase of growth was similar in cells cultivated at the different temperatures. No difference in cDA (between 0.11 and 1.87 pg DA cell⁻¹) was observed in cells cultivated at the two nutrient ratios. To evaluate if P. australis accumulates DA (cDA + dDA) at different stages of the culture and not only during the stationary phase of growth, the BTS-1 strain was cultivated at 14 °C and the content of this toxin was measured during culture development. The cultures were maintained at high (HL; 200 μmol quanta m⁻² s⁻¹) and low light (LL; 30 μmol quanta m⁻² s⁻¹) and in the two nutrient ratios to evaluate the effect of these variables on DA content. The photosynthetic performance and pigment concentration were measured as indicators of the physiological condition of the cells. cDA was detected in all culture conditions and during the different stages of growth. The highest DA content was measured during the lag phase of growth and it was present mainly in the medium (dDA = 70.83 pg DA cell⁻¹). Cells cultivated at HL produced more DA than LL cultured cells. P. australis cultured in HL presented lower photosynthetic rates than LL cells and had similar concentrations of photoprotective pigments and the highest maximum photosynthetic rates were detected during the lag phase of growth in all culture conditions. The results demonstrate that P. australis from northern Baja California peninsula presents a narrow temperature range for optimal growth under batch culture conditions. P. australis produce DA at different stages of growth, and DA content was related to the light intensity at which the cells were cultivated.     

Thermoregulation on the air–water interface—II: Foot conductance,activity metabolism and a two-dimensional heat transfer model

We used a quasi-adiabatic calorimeter and respirometry apparatus to measure heat loss from the feet of 3- to 4-d-old mallard ducklings (Anas platyrhynchos). We found that, at cool (<20 °C) operative temperatures, foot conductance increased in proportion to operative temperature, T_e, rather than water temperature. We combined these results with those of an earlier study to develop a heat transfer model for swimming ducklings. This model includes separate thermal conductances to air (0.027 W/°C-animal), to water through the down (0.035[1+2.05×10⁻⁷T_e⁴]) W/°C-animal, and to water through the feet (2.01×10⁻⁸T_e⁴ W/°C-animal). The overall conductance by all three routes is only 21% greater when swimming compared to standing in air at the same operative temperature. Interestingly, ducklings can maintain body temperature >39 °C while swimming in 5 °C water, but not when restrained in a calorimeter with 5 °C water. Peak oxygen consumption is greater when swimming, and apparently exercise metabolism substitutes almost completely for thermoregulatory heat production.     

The effects of light intensity on the growth of Japanese Gambierdiscus spp. (Dinophyceae)

Marine toxic dinoflagellates of the genus Gambierdiscus are the causative agents of ciguatera fish poisoning (CFP), a form of seafood poisoning that is widespread in tropical, subtropical and temperate regions worldwide. The distributions of Gambierdiscus australes, Gambierdiscus scabrosus and two phylotypes of Gambierdiscus spp. type 2 and type 3 have been reported for the waters surrounding the main island of Japan. To explore the bloom dynamics and the vertical distribution of these Japanese species and phylotypes of Gambierdiscus, the effects of light intensity on their growth were tested, using a photoirradiation-culture system. The relationship between the observed growth rates and light intensity conditions for the four species/phylotypes were formulated at R > 0.92 (p < 0.01) using regression analysis and photosynthesis-light intensity (P-L) model. Based on this equation, the optimum light intensity (L_max) and the semi-optimum light intensity range (L_s-opt) that resulted in the maximum growth rate (μ_max) and ≥80% μ _max values of the four species/phylotypes, respectively, were as follows: (1) the L_max and L_s-opt of G. australes were 208 μmol photons m⁻² s⁻¹ and 91–422 μmol photons m⁻² s⁻¹, respectively; (2) those of G. scabrosus were 252 and 120–421 μmol photons m⁻² s⁻¹, respectively; (3) those of Gambierdiscus sp. type 2 were 192 and 75–430 μmol photons m⁻² s⁻¹, respectively; and (4) those of Gambierdiscus sp. type 3 were ≥427 and 73–427 μmol photons m⁻² s⁻¹, respectively. All four Gambierdiscus species/phylotypes required approximately 10 μmol photons m⁻² s⁻¹ to maintain growth. The light intensities in coastal waters at a site in Tosa Bay were measured vertically at 1 m intervals once per season. The relationships between the observed light intensity and depth were formulated using Beer’s Law. Based on these equations, the range of the attenuation coefficients at Tosa Bay site was determined to be 0.058–0.119 m⁻¹. The values 1700 μmol photons m⁻² s⁻¹, 500 μmol photons m⁻² s⁻¹, and 200 μmol photons m⁻² s⁻¹ were substituted into the equations to estimate the vertical profiles of light intensity at sunny midday, cloudy midday and rainy midday, respectively. Based on the regression equations coupled with the empirically determined attenuation coefficients for each of the four seasons, the ranges of the projected depths of L_max and L_s-opt for the four Gambierdiscus species/phylotypes under sunny midday conditions, cloudy midday conditions, and rainy midday conditions were 12–38 m and 12–54 m, 1–16 m and 1–33 m, and 0 m and 0–16 m, respectively. These results suggest that light intensity plays an important role in the bloom dynamics and vertical distribution of Gambierdiscus species/phylotypes in Japanese coastal waters.     

An LED-based UV-B irradiation system for tiny organisms: System description and demonstration experiment to determine the hatchability of eggs from four Tetranychus spider mite species from Okinawa

We developed a computer-based system for controlling the photoperiod and irradiance of UV-B and white light from a 5 × 5 light-emitting diode (LED) matrix (100 × 100 mm). In this system, the LED matrix was installed in each of four irradiation boxes and controlled by pulse-width modulators so that each box can independently emit UV-B and white light at irradiances of up to 1.5 and 4.0 W m⁻², respectively, or a combination of both light types. We used this system to examine the hatchabilities of the eggs of four Tetranychus spider mite species (T. urticae, T. kanzawai, T. piercei and T. okinawanus) collected from Okinawa Island under UV-B irradiation alone or simultaneous irradiation with white light for 12 h d⁻¹ at 25 °C. Although no eggs of any species hatched under the UV-B irradiation, even when the irradiance was as low as 0.02 W m⁻², the hatchabilities increased to >90% under simultaneous irradiation with 4.0 W m⁻² white light. At 0.06 W m⁻² UV-B, T. okinawanus eggs hatched (15% hatchability) under simultaneous irradiation with white light, whereas other species showed hatchabilities <1%. These results suggest that photolyases activated by white light may reduce UV-B–induced DNA damage in spider mite eggs and that the greater UV-B tolerance of T. okinawanus may explain its dominance on plants in seashore environments, which have a higher risk of exposure to reflected UV-B even on the undersurface of leaves. Our system will be useful for further examination of photophysiological responses of tiny organisms because of its ability to precisely control radiation conditions.     

Varying endophyte status and energy supplementation of fresh tall fescue in continuous culture

Eight dual-flow continuous culture vessels (700 ml) were used to compare in vitro effects of toxic, endophyte-infected (E+), endophyte-free (E−), and non-toxic, endophyte-infected (EN) Jesup tall fescue (vegetative stage) on ruminal fermentation at 4 levels (0, 150, 300, and 450 g kg⁻¹ DM) of concentrate supplementation (ground corn) for a total of 12 experimental diets in a randomized incomplete block design with 2 replicates. Each culture vessel was offered a total of 15 g DM d⁻¹. Forage was fed in four equal portions (fed at 03:00, 09:00, 15:00, and 21:00 h); and corn was fed in two equal portions (fed at 09:00 and 21:00 h). Headspace gas and liquid samples were analyzed for methane, ruminal culture pH, ammonia–N, and volatile fatty acid production. Ammonia–N output (g d⁻¹) varied by grass; EN had lower values compared to those of E+ and E−. Increasing the level of grain linearly decreased ruminal culture pH, ammonia–N, acetate production, and the acetate-to-propionate ratio, whereas propionate and butyrate production increased with higher grain supplementation. Ruminal fermentation was minimally altered by the presence of the endophyte; however, for the highest level of grain fed (450 g kg⁻¹ DM fed) the methane production pattern for all three grasses was altered. In addition to having the lowest ruminal ammonia–N accumulation, the non-toxic, endophyte-infected fescue resulted in the lowest methane production measured.