Nutrient-induced competition between two species of marine diatoms

Competition experiments betweenPhaeodactylum tricornutum andSkeletonema costatum showed that even at temperatures higher than 10°C (i.c. 14°C), the development ofSkeletonema can be favoured by adjusting nutrient levels and nutrient ratios. Low NSi ratios were found to favourSkeletonema. Additionally, high NP ratios further enhanced the ability ofSkeletonema to dominate the cuftures. Contrary to some statements in literature, it seems that high concentrations of silicates are more important for the dominance ofSkeletonema costatum in large-scale cultures than just low temperatures. This finding is important with regard to stimulating the blooming ofSkeletonema costatum in natural phytoplankton populations as food for bivalve molluscs.     

Effects of light,temperature and salinity on the growth rate of harmful marine diatoms,Chaetoceros convolutus and C. concavicornis that kill netpen salmon

Chaetoceros convolutus and C. concavicornis have been implicated in the death of salmon in netpens in the Pacific Northwest by damaging the salmon's gills. To better understand how environmental factors affect the distribution of these two species, the interacting effects of light, temperature and salinity on growth rate were examined by growing these species under a range of temperatures (4–18 °C), light (10–175 μmol photon m⁻² s⁻¹) and salinities (10–30‰). For C. convolutus, the growth rate showed a hyperbolic relationship with irradiance at 8, 14 and 18 °C and light saturation occurred at 9, 14 and 20 μmol photon m^t s⁻¹ respectively. At 4 °C for C. convolutus and 8 °C for C. concavicornis, cells grew at μ_max, even at the lowest irradiances tested (10 μmol photon m⁻² s⁻¹). For C. convolutus, the amount of light required to saturate growth rate increased with temperature in an approximately linear fashion. The Q₁₀ was 1.88, calculated by averaging over both species. C. concavicornis was the more euryhaline species growing at salinities as low as 17.5‰, while C. convolutus grew only at 25‰ and above.     

Growth physiology of a marine nitrogen-fixing cyanobacterium (Nodularia harveyana) in outdoor culture

The performance ofNodularia harveyana, a N₂-fixing cyanobacterium isolated from seawater, has been studied outdoors in two different culture systems: open pond (OP) and tubular photobioreactor (TPR). The productivity in both devices was influenced by areal density. The maximum yield obtained was 12.0 g (d.wt) m^–2 day^–1 in OP and 14.0 g (d.wt) m^–2 day^–1 in TPR in August, corresponding to the highest solar radiation received. In a month-long experiment with the cyanobacterium cultivated in TPR at high circulation speed, a net increase in productivity was obtained over that at low circulation speed. The influence of temperature on the productivity of the cultures grown in open ponds and tubular photobioreactors has been investigated. The higher productivity obtained in TPR compared to OP was attributed to its better controlled temperature conditions. In outdoor culture the maximum nitrogenase activity did not coincide with the maximum light intensity, but occurred in early afternoon. The amount of carbohydrate accumulated during the day probably influenced the rate of dark nitrogenase activity and its duration in the night.     

Reproductive isolation among sympatric cryptic species in marine diatoms

Pseudo-nitzschia is a marine cosmopolitan genus of chain-forming planktonic diatoms. As for the vast majority of phytoplankton organisms, species identification within this genus mostly relies upon morphological features. Taxa were initially identified based on cell shape and gross morphology of their composite silica cell wall, called the frustule. Yet, observations of the frustule in electron microscopy showed many additional characters for species identification and results of molecular studies have demonstrated that genetically distinct groups might exist within morpho-species. However, these studies have not addressed the biological meaning of these genetic differences. Here, we bridge that gap by comparing ultrastructural features and sequence data (three ribosomal and one plastid marker) of 95 strains with results of mating experiments among these strains. Experiments were performed on two morphologically distinct entities: P. delicatissima and P. pseudodelicatissima. Each of the two entities consisted of multiple genetically distinct and reproductively isolated taxa, all occurring in sympatry: P. delicatissima was composed of three phylogenetic and reproductively distinct groups, whereas P. pseudodelicatissima consisted of up to five. Once these taxa had been defined both genetically and biologically, subtle ultrastructural differences could be detected as well. Our findings not only show that cryptic genetic variants abound in sympatry, but also that they are reproductively isolated and, therefore, biologically distinct units.     

Agar yield and quality of Gracilaria chilensis (Gigartinales,Rhodophyta) in tank culture using fish effluents

Tank cultivation of Gracilaria using fish effluents has permitted a production of 48 kg m^–2 yr^–1 and can reduce the dissolved nitrogen loads in the seawater. We report the yield, gel strength, gelling and melting point of agar from Gracilaria cultivated in tanks with seawater previously utilized in intensive, land-based salmon cultures and compared to a control using directly pumped seawater, over a study period of 22 months. The results show that the highest agar yield (20 to 22%) was obtained when Gracilaria was cultivated with pure seawater as compared to the fish effluents. The gel strength, gelling and melting point were higher in the agar obtained from algae cultured with fish effluents. During the spring, the gel strength, gelling and melting point increased in tanks with fish effluents and decreased in tanks with a supply of pure seawater.     

Changes in fatty acid profiles of thermo-intolerant and thermo-tolerant marine diatoms during temperature stress

Fatty acid composition and degree of fatty acid saturation during temperature stress in thermo-intolerant (Phaeodactylum tricornutum) and thermo-tolerant (Chaetoceros muelleri) marine diatoms were investigated. A greater number of fatty acids were observed in C. muelleri than in P. tricornutum regardless of treatment. The major fatty acids detected were 14:0, 16:0, 16:1, 16:2, 16:3, 18:0, 18:1(n-9)c, 18:2(n-6) and 20:5(n-3) with additional fatty acids 18:1(n-9)t and 20:4(n-6) detected in C. muelleri. Short duration (2 h) temperature increase above optimal growth temperature had a greater effect on fatty acid composition in C. muelleri than in P. tricornutum and the degree of fatty acid saturation was affected more by temperature in C. muelleri than in P. tricornutum during both short and long duration (24 h) treatments. Total protein assay results suggest that P. tricornutum, but not C. muelleri, was undergoing stress under our growing conditions although lipids in both diatoms were affected by increased temperature. Immunodetection of proteins with anti-rubisco indicates that the rubisco large subunit was undergoing greater turnover in C. muelleri than in P. tricornutum. However, the integrity of rubisco as a suitable indicator of lipid status needs further study. This work supports the hypothesis that a particular temperature, and not treatment duration, has the greater effect on changes in fatty acid composition. Furthermore, changes in fatty acid composition and degree of fatty acid saturation occurred more quickly in the diatoms in response to increased temperature than previously observed in nutrient starvation studies. Since diatom lipids represent an important resource for growth and reproduction of marine animals, the rapid alteration of their lipid composition under temperatures normally encountered in marine environments warrants further study.     

Productivity analysis of outdoor chemostat culture in tubular air-lift photobioreactors

Net productivity and biomass night losses in outdoor chemostat cultures ofPhaeodactylum tricornutum were analyzed in two tubular airlift photobioreactors at different dilution rates, photobioreactor surface/volume ratios and incident solar irradiance. In addition, an approximate model for the estimation of light profile and average irradiance inside outdoor tubular photobioreactors was proposed. In both photobioreactors, biomass productivity increased with dilution rate and daily incident solar radiation except at the highest incident solar irradiances and dilution rates, when photoinhibition effect was observed in the middle of the day. Variation of estimated average irradiance vs mean incident irradiance showed two effects: first, the outdoor cultures are adapted to average irradiance, and second, simultaneous photolimitation and photoinhibition took place at all assayed culture conditions, the extent of this phenomena being a function of the (incident)¹ irradiance and light regime inside the culture. Productivity ranged between 0.50 and 2.04 g L^–1 d^–1 in the tubular photobioreactor with the lower surface/volume ratio (S/V = 77.5 m^–1) and between 1.08 and 2.76 g L^–1 d^–1 in the other (S/V = 122.0 m^–1). The optimum dilution rate was 0.040 h^–1 in both reactors. Night-time biomass losses were a function of the average irradiance inside the culture, being lower in TPB0.03 than TPB0.06, due to a better light regime in the first. In both photobioreactors, biomass night losses strongly decreased when the photoinhibition effect was pronounced. However, net biomass productivity also decreased due to lower biomass generation during the day. Thus, optimum culture conditions were obtained when photolimitation and photoinhibition were balanced.     

A model of seaweed growth in an outdoor culture in Israel

The agarophyte red seaweed Gracilaria conferta was used as a model plant to study the relationship between growth and environmental factors. This species was cultured in small outdoor tanks with continuous seawater supply during three years. Seaweeds were kept under constant density by weekly thinning and were also pulse-fed weekly with nitrogen and phosphate. Water temperature and underwater irradiance increase had opposite effects on the weekly growth rate in two seasons: negative in summer and positive in non-summer. Therefore, a dichotomic separation between summer (June–August) and non-summer (September–May) seasons was utilized in the proposed linear model. The linear model, of the analysis of covariance type, accounted for an explained percentage of total variation (R²) of 0.567, with significant coefficients of all variables included. A standardized model showed that season was the dominant variable, with its coefficient being twice that of temperature in summer, and zero in the non-summer season. Water temperature affected the growth rate twice as much as irradiance, and epiphytes showed a significant negative effect on growth only in the summer. This model aids in the prediction of growth on a seasonal basis under local conditions.     

Restoration of Botshol (The Netherlands) by reduction of external nutrient load: The effects on physico-chemical conditions,plankton and sessile diatoms

The nature reserve Botshol (Utrecht, The Netherlands), consisting of two shallow lakes, ditches and reedland, originated from excavation of peat by man in the 17th century. Up to 1960 Botshol was a clear-water Charophyte lake system. Since the sixties water quality deteriorated and phytoplankton concentrations increased, while the number and dispersion of Chara species decreased. Several restoration measures were attempted to restablish a Charophyte-dominated ecosystem. This paper reports the promising results of this restoration experiment and mentions some complications that arose in restoring the reserve to a less fertile state. The restoration measures have resulted in a sixfold reduction of the external phosphorus load, from 0.6 to 0.1 g m^–2.y^–1, and in a significant reduction of phosphorus levels at all locations. Moreover, the light climate improved and the phyto- and zooplankton compositions changed considerably Unexpectedly, a bloom ofPrymnesium parvum and a fish kill were observed during the last three months of 1990. Despite this fish kill the restoration of the lake is successful so far.     

Effect of blue-green light on growth rate and chemical composition of three diatoms

The diatomsChaetoceros sp.,Skeletonema costatum andThalassiosira pseudonana were grown with different irradiances of white and of blue-green light, and with a mixture of blue-green plus 6.5 mol m^–2 s^–1 of white light. Exponential growth rates were higher in mixed blue for the first two, whileT. pseudonana grew faster in white light but, in all cases, mean cell division rates did not differ with increasing irradiances. Harvesting in stationary, rather than in late exponential growth phase, resulted in higher protein contents forChaetoceros sp. andS. costatum, but forT. pseudonana the highest value was in the exponential phase. The highest protein content was in blue-green light for the three species and it increased with irradiance. As to other fractions, the three strains showed different responses, related to quality and quantity, as well as to culture ages.     

Feeding by marine fish larvae: developmental and functional responses

Synopsis The relationship between prey consumption rate and prey concentration (functional response), and its change with growth (developmental response) were examined in the laboratory for three species of marine fish larvae: bay anchovy Anchoa mitchilli (Engraulidae), sea bream Archosargus rhomboidalis (Sparidae) and lined sole Achirus lineatus (Soleidae). The major objective was to determine relative predatory abilities of the larvae by fitting feeding rate data to developmental and functional response models. Feeding success, prey capture success, attack rates, handling times and search rates were estimated. Prey consumption rates and attack rates of bay anchovy usually were highest, but at the lowest prey level (50 per liter) first-feeding sea bream larvae had the highest consumption rate. Sea bream could consume prey at near-maximum rates at prey levels lower than those required by the other species. As larvae grew, time searching per attack decreased rapidly for all species, especially at low prey levels. Handling time also decreased, but most rapidly for bay anchovy. Search rates were highest for bay anchovy and lowest for lined sole. Bay anchovy had the best apparent predation ability, but when previous results on larval growth rates, survival rates and growth efficiencies were considered, sea bream larvae were the most efficient predators and the least likely of the three species to be limited by low prey levels.     

Screening of diatoms for heterotrophic eicosapentaenoic acid production

Nine strains of diatoms (representing four species) were screened for their ability to produce eicosapentaenoic acid (EPA) when cultured heterotrophically on glucose. Four strains were able to produce EPA heterotrophically using glucose as its carbon and energy source. Of the four,Nitzschia laevis was the best EPA producer, yielding 0.017 g g^–1 dry cell weight.N. laevis was the only species tested which synthesised more EPA heterotrophically than photosynthetically. This study shows thatN. laevis is a potential source of EPA production using heterotrophic culture conditions with glucose as the carbon and energy substrate.Author for correspondence     

Experiments on Spirulina culture on waste-effluent media and at the pilot plant

The prevalence of the microsporidian, Nosema herpobdellae, in the leech, Erpobdella octoculata, collected from a eutrophic English lake, was examined at monthly intervals over two years. The leech has an annual life cycle and, for each cohort, infection appeared in recently recruited animals in early autumn and prevalence levels rose to a peak in late autumn or winter before declining till the cohort died out the following autumn. Prevalence levels never exceeded 9%. The effect of the parasite on young leeches was difficult to assess, but there was some evidence of parasite-induced host mortality in older animals.     

A direct immunoassay for detecting diatoms in groundwater as an indicator of the direct influence of surface water

Groundwater under the direct influence of surface water (GWUDISW) is of concern in communities where growing public demand on groundwater resources has resulted in increased withdrawals and hydraulic stress near surface water bodies. Under these conditions, contaminants such as methyl-tert butyl ether (MTBE) and biological materials have been detected in domestic wells. Other contaminants and pathogens associated with surface water are not routinely tested for in groundwater-supplied systems. To address the need for methods to easily identify potentially vulnerable supplies, a direct immunoassay for the quantitative detection of diatoms in raw water samples was developed as a measure of surface water influence on groundwater. Cell wall preparations from Nitzschia palea Kützing, a freshwater diatom found throughout North America, were used to produce a polyclonal antibody that was applied in a direct enzyme-linked immunosorbent assay (ELISA) developed to detect the presence of N. palea cell wall components. The direct immunoassay allows detection at 500 cells L⁻¹, a level similar to diatom concentrations observed in samples of groundwater collected near the test site. This investigation was the first attempt to utilize an ELISA as an indicator of surface water influence on groundwater. Further research is needed to develop more specific diatom-based monoclonal antibodies, determine cross-reactivity, and optimize sample processing and ELISA procedures for development of a standardized method.     

Ingestion of live filamentous diatoms by the Great Lakes amphipod,Diporeia sp.: a case study of the limited value of gut contents analysis

Individuals of the Great Lakes amphipod, Diporeia sp. (formerly named Pontoporeia hoyi) were collected from a 45-m deep station in southeastern Lake Michigan and isolated in small laboratory feeding vessels at 4 °C, after the animals had voided their guts over a 24-hour period. Over a 20-day period, following introduction of a single ration of live cells of the filamentous diatom, Melosira varians, 9 of 10 animals had ingested this material, and 7 of these 9 individuals had deposited fecal pellets. Subsequent examination of gut contents and fecal pellets showed that although animals had ingested whole algal cells/filaments, little of the material in gut contents or fecal pellets bore any identifiable structural similarity to cells/filaments prior to ingestion. The results suggest that earlier studies of pontoporeiid gut contents may seriously underestimate the importance of algal components in the amphipod's diet and imply that Diporeia sp. growth and production may be more closely linked to primary production than previously thought.     

Weather-driven ecology of planktonic diatoms in Lake Tovel (Trentino,Italy)

The seasonal development of planktonic diatoms in Lake Tovel has been studied from 2002 to 2004 within the SALTO research project. The objective was to investigate the role of weather conditions and lake hydrology in regulating the development of Cyclotella and Fragilaria functional species groups. The different responses of the two species to the environmental variability and their reciprocal relationships were considered. Cyclotella showed a weak relationship with the selected environmental variables and its seasonal development was difficult to interpret, also in relation to its opportunistic behaviour. On the contrary, Fragilaria complex showed a prompt response to environmental variability and its seasonal development appeared to be mainly regulated by hydrological conditions, through nitrate nitrogen and, secondarily, by silica concentrations. Water temperature and thermal stability affected only Fragilaria species, while different nutrient requirements allowed Cyclotella species to grow under conditions of limited resources for Fragilaria. Since Lake Tovel has scarce internal nutrient reserves and its hydrology is strongly affected by precipitation both in winter and summer, diatom phenology seems to be indirectly regulated by weather conditions.     

In vitro culture of marine trematodes from their snail first intermediate host

The ability to culture parasites outside their host (i.e. in vitro) is essential for several aspects of parasitological research. Here, a culture medium for marine trematode parthenitae was optimized using Philophthalmus sp. rediae from the intermediate snail host, Zeacumantus subcarinatus. The medium was optimized by sequentially testing the suitability of different levels of osmolality, different commercially available media, and different concentrations of supplemented chicken serum, while controlling for genetic variation among cultures. Philophthalmus sp. rediae survived up to 56 days in cultures of the best tested medium, remaining active and continuously shedding cercariae. The broader suitability of the culture medium was tested using five other trematode species from different families (using either the same or other marine snails as first intermediate hosts): Galactosomum sp., Acanthoparyphium sp., Maritrema novaezealandensis, Curtuteria australis, and an undescribed species of the family Opecoelidae. Survivorship of rediae and sporocysts from these species ranged from eight days to 42 days. The culture procedures developed here can therefore be used in the future as a system under which to culture marine trematode parthenitae for experimental studies.     

Effect of UV radiation on photoinhibition of marine macrophytes in culture systems

The present study examined the effect of UV andphotosynthetically active radiation (PAR) onphotoinhibition and recovery in the Phaeophyte Macrocystis pyrifera, the Rhodophyte Chondruscrispus and the Chlorophyte Ulva lactuca underoutdoor culture conditions. There was an increase inphotoinhibition as a consequence of high exposure toUV-B radiation in M. pyrifera, however, highlevels of PAR accounted for most of thephotoinhibition in C. crispus and U.lactuca. Photodamage by UV-A, UV-B and PAR wascompletely repaired within 5 h and effective quantumyield reached pretreatment values in the three speciesstudied. Species were less susceptible tophotoinhibition after being incubated for 5 d underhigh exposures of natural irradiance suggesting aphotoadaptive process. The recovery of the effectivequantum yield was impaired by long exposure to highlevels of UV-B in C. crispus and UV-A, UV-B andPAR in M. pyrifera. This suggests a differentkind of damage by UV-A and PAR radiation, one to thephotosynthetic apparatus and another which affects therepair mechanism of some species. There was anincrease in UV-absorption ( 330 nm) in M. pyrifera and C. crispus within four days ofthe initiation of the experiment suggesting that thesespecies photoprotect their photosynthetic system whenexposed to elevated UV and PAR levels.     

Variation of fatty acid profile with solar cycle in outdoor chemostat culture ofIsochrysis galbana ALII-4

Outdoor chemostat cultures of the marine microalgaIsochrysis galbana at constant dilution rate (0.034 h^–1 ) have been carried out under different weather conditions. Steady-state biomass concentrations were 1.61±0.03 kg m^–3 in May and 0.95±0.04 kg m^–3 in July, resulting in biomass output rates of 0.54 kg m^–3 d^–1 and 0.32 kg m^–3 d^–1 in May and July, respectively. Two patterns of daily variation with the solar cycle were observed in the fatty acid content. Saturated and mono-unsaturated fatty acids (16:0 and 16:1n7) show significant variation with the solar cycle, associated with short-term changes in environmental factors. Palmitic and palmitoleic acids are generated during daylight and consumed during the dark period. However, polyunsaturated fatty acids do not show a significant response to the solar cycle and their changes are associated with long-term variation in environmental factors. The maximum EPA productivity was obtained in May, 14.1 g m^–3 d^–1, which is close to that found in the literature for indoor continuous cultures. Nonetheless, the outdoor EPA content (up to 2.61 % d.wt) was lower than the indoor EPA content from a previous study (5% d.wt).