Biochemical composition of crustacean zooplankton and their grazing on phytoplankton and ciliated protozoans in a recently founded reservoir (Sahela, Morocco

In order to assess the impact of crustacean zooplankton on phytoplankton and protozoan ciliates in the Sahela reservoir under semi-arid climate, we conducted experiments during the period from July to December 1999 at the deepest point in the lake (15 m). Samplings and measurements were carried out in diffusion chambers submerged in situ over a period of 7 h without (control chambers) and with (experimental chambers) crustacean zooplankton. During these experiments, counts were conducted on phytoplankton and ciliates to determine the abundance and the mortality of these organisms due to zooplankton in each diffusion chambers at t = 0 and t = 7 h of incubation. The study showed that the growth rates of phytoplankton and ciliates populations varied between 0.02 and 0.05 h-1 and from 0.01 to 0.07 h-1, respectively. The mortality caused by zooplankton grazing fluctuated from 0.07 to 0.2 h-1 of phytoplankton and from 0.01 to 0.2 h-1 of ciliates. These mortalities were significantly and positively correlated with the growth rates (r = 0.8; p < 0.02; n = 9). Moreover, the heavy predation by the crustacean zooplankton was exerted on small-sized phytoplankton and ciliates and we demonstrated the relationships between protozoans and zooplankton for the transfer of matter and energy in aquatic food webs. Furthermore, the crustacean zooplankton metabolism was different, whether zooplankton was present in diffusion chambers or in the lake.     

Manipulation of dietary lipids, fatty acids and vitamins in zooplankton cultures

1. A wide range of species that are cultivated in commercial mariculture are planktonic during at least part of their life cycle; for example, the larval stages of shellfish (shrimp and molluscs) and the live feeds (rotifers, brine shrimp, copepods) used in the larviculture of marine fish and shellfish. Over the last decades various techniques have been developed to deliver nutrients to these zooplanktonic organisms either through artificial diets or by manipulating the composition of the live prey fed to the carnivorous stages. This paper reviews the methodology that has allowed aquaculturists to gain knowledge of nutritional requirements and may offer interesting opportunities for ecologists to verify the importance of key nutrients in the natural food chain of marine as well as freshwater ecosystems.
2. Live micro-algae can be replaced partially or completely in the diet of filter-feeders such as rotifers, Artemia , shrimp larvae and bivalves, by various types of preserved algae, micro-encapsulated diets and yeast-based diets, whereas lipid emulsions and liposomes may be utilized to supplement specific lipid-and water-soluble nutrients, respectively. Microbound and micro-encapsulated diets have been designed to supplement live feed in the culture of micro-predators such as fish and shrimp larvae.
3. Live prey organisms, in particular rotifers and Artemia , can be 'bio-encapsulated' with a variety of enrichment diets to manipulate their content in certain nutrients, including ω3 highly unsaturated fatty acids (FA) and the vitamins C, A and E. Nevertheless, the enrichment techniques are not applicable for all nutrients and prey organisms. Phospholipid composition is difficult to manipulate through the diet of live feed and the enrichment of the essential FA docosahexaenoic acid (DHA) is hampered in most Artemia species due to the catabolism of this FA following enrichment.     

Dissolved organic carbon released by zooplankton grazing activity-a high-quality substrate pool for bacteria

Experiments were designed to investigate whether processes relatedto zooplankton feeding have a positive effect on bacterial growth.Bacterial abundance and [³H]thymidine incorporation rates werefollowed in grazer-free batch cultures originally containingeither Scenedesmus quadricauda or Rhodomonas lacustris as foodsources, and Daphnia cucullata or Eudiaptomus graciloides asgrazers. Compared with controls lacking either animals or algae,a significantly higher bacterial abundance and productivityoccurred in cultures which contained both phyto- and zoo-plankton.The same experimental methodology was tested during the declineof a diatom spring bloom in a eutrophic, temperate lake. A significantincrease in bacterial biomass was observed due to the grazingactivity of in situ mesozooplankters during the clear-waterphase. Our results demonstrated that the dissolved carbon pathwayfrom mesozooplankton to bacteria averaged 57% (26–78%)of the algal carbon filtered from suspension.     

Effects of rare earth elements on growth rate,lipids, fatty acids and pigments in microalgae

The extensive exploitation of rare earth elements (REEs), particularly in electronic technologies and agriculture has concomitantly raised the environmental load. Their resulting effects on primary producers such as microalgae are, however, poorly understood. We have studied these effects on two microalgae of biotechnological interest. The yellow‐green alga Trachydiscus minutus (Eustigmatophyceae, Ochrophyta), and the green alga Parachlorella kessleri (Trebouxiophyceae, Chlorophyta) were cultivated in mineral medium supplemented with 10 μmol L^?1 chlorides of REEs: cerium, gadolinium, lanthanum, lutetium, praseodymium, scandium, and with monazite, which is a mineral rich in those elements. We observed growth rates at different mean light intensities (20, 50, 150 and 300 μmol m^?2 s^?1). The high growth rate of P. kessleri was not affected by the presence of any lanthanide, and decreased proportionally with light intensity (from 0.2 to 0.04 doublings per hour). In contrast, the growth rate of T. minutus was about three times lower compared with P. kessleri, with an optimum at 50 μmol m^?2 s^?1 and decreased at higher or lower light intensities. In the presence of Ce³⁺, La³⁺ and Sc³⁺, the growth rate markedly increased to a value that corresponded to the growth rate in P. kessleri at the same light intensity. The composition and content of pigments and lipids were followed at the optimum light intensity for both species. The lipid content (percentage of dry weight) varied only slightly in the presence of individual rare earths. There was, however, an increase in saturated fatty acids at the expense of polyunsaturated fatty acids. The effect of REEs on pigments was variable: the presence of Ce³⁺, Gd³⁺, La³⁺ and Sc³⁺ caused an increase in the concentrations of major pigments such as lutein, violaxanthin, β‐carotene or chlorophylls, while Pr³⁺ and Lu³⁺ reduced them.     

Short-term productivity responses of algae and bacteria to zooplankton grazing in two freshwater lakes

SUMMARY. 1. The specific productivities of algae and bacteria were measured in short-term (4 day) experiments consisting of enclosures with natural or reduced zooplankton biomass. Experiments were repeated five times over a season in each of two lakes that differed in the background concentration of dissolved organic carbon (DOC).
2. Algal biomass as estimated by chlorophyll a was suppressed in enclosures with ambient grazer levels in six of ten experiments and enhanced in one experiment. Distribution of chlorophyll among net and nanoplankton was not significantly affected by grazing.
3. Relative to enclosures with reduced zooplankton, normal grazer biomass (97–466μg 1⁻¹ dry weight) enhanced specific algal productivity in only one of five experiments in the low DOC take and had no effect in all five experiments in the high DOC lake. The main effects of grazers on algae was through removal of biomass rather than through indirect changes in turnover rate.
4. Between experiments, bacterial density was either unaffected, or mildly enhanced (4–87%) in enclosures with ambient macrozooplankton compared to those with reduced levels. Bacterial productivity and turnover estimated by incorporation of [³H]thymidine into DNA showed different responses across experiments; increasing, declining or remaining the same with grazer minipulation. This variability was not related to differences in dissolved primary production or to background DOC between lakes or experiments. Comparison of bacterial productivities based on thymidine incorporation rates with changes in cell densities indicated that control of bacterial loss processes by macrozooplankton is more important than control of growth rates.     

Effect of short-chain fatty acids on the size of enteric bacteria

J.L. THOMPSON AND M. HINTON. 1996. The size of Escherichia coli and Salmonella enteritidis PT4 cells was measured using both transmission electron micrographs and image analysis. Incubation in the presence of formic and propionic acids resulted in larger cells, possibly as a result of DNA inhibition, with no apparent damage to the cell membranes. Bacteria incubated with propionic acid were more resistant to shrinkage after fixation, possibly as a result of altered phospholipid and fatty acid composition.     

FO-spectra of chlorophyll fluorescence for the determination of zooplankton grazing

In the PHYTO-PAM phytoplankton analyzer the minimal fluorescence of dark-adapted samples (F₀) was assessed, which gives direct information on the chlorophyll-a content. Clearance rates (CR) of Daphnia and Brachionus were calculated from a decrease in chlorophyll-a concentration using the PHYTO-PAM fluorometer for non-sacrificial sampling of chlorophyll-a. Clearance rates of Daphnia were measured and compared with those based on the cell-counts method using an electronic particle counter (Coulter counter). Chlorophyll fluorescence-based CR for Daphnia magna were very strongly correlated with Coulter-based CR, signifying the potential suitability of the PHYTO-PAM in grazing experiments. A procedure for determination of rotifer clearance rates was developed and the effects of rotifer density, duration of the grazing period, and food concentration on CR were investigated. Between 10 and 30 rotifers in 2.5 ml food suspension (i.e. 4–12 rotifers per ml) appeared optimal for calculating CR. The application of the deconvolution of F₀-spectra in food selectivity experiments was evaluated using various mixtures of the green alga Scenedesmus obliquus and the cyanobacterium Microcystis aeruginosa fed to Brachionus. CR for Brachionus on M. aeruginosawere lower than on S. obliquusbut this was not caused by toxicity, because no mortality was observed. The higher CR on Scenedesmus than on Microcystis in the mixtures suggested selectivity. The importance of digital suppression of background fluorescence is highlighted in additional experiments with Daphnia feeding on mixtures of Microcystis and Scenedesmus, or on Microcystis alone. Without background correction of filtered samples, negative clearance rates were obtained for the `blue' Microcystis signal. Soluble fluorescing compounds of cyanobacterial origin, phycocyanin, were released from the Daphniaand contributed 40% to the overall-fluorescence. Deconvolution of F₀-spectra for the determination of chlorophyll-a using the PHYTO-PAM appears to be a suitable tool for determination of rotifer CR even at very low food concentrations. A drawback of the method is that rather high rotifer densities are required. The required grazing period, however, is shorter than for cell-count methods, the method is sensitive, clearance rates can be measured at low food concentrations (< 0.1 mg C l^–1) and information on selective feeding can be obtained.     

A field technique for the determination of zooplankton grazing on natural bacterioplankton

SUMMARY. 1. A method for the determination of zooplankton community grazing rates on natural populations of bacteria is described.
2. Undisturbed zooplankton samples were incubated with ³H-thymidine-labelled bac teriioplankton, and animals and their food subsequently separated by size.
3. Prehandling of zooplankton by concentration before incubation reduce the grazing rates by a factor ranging from three to four.
4. Time series experiments showed that appropriate incubation periods ranged from 40 min at 14°C to 60 min at 10°C.
5. Spatial variability in the zooplankton was the main source of the overall standard error. This was considerably reduced by using composite samples.     

Lipase-catalyzed esterification of glycerol and polyunsaturated fatty acids from fish and microalgae oils

This paper reports on the synthesis of triglycerides by enzymatic esterification of polyunsaturated fatty acids (PUFA) with glycerol. The lipase Novozym 435 (Novo Nordisk, A/S) from Candida antarctica was used to catalyze this reaction. The main factors influencing the degree of esterification and triglyceride yield were the amount of enzyme, water content, temperature and glycerol/fatty acid ratio. The optimum reaction conditions were established as: 100 mg of lipase; 9 ml hexane; 50°C; glycerol/PUFA concentrate molar ratio 1.2:3; 0% initial water; 1 g molecular sieves added at the start of reaction; and an agitation rate of 200 rpm. Under these conditions, a triglyceride yield of 93.5% was obtained from cod liver oil PUFA concentrate; the product contained 25.7% eicosapentaenoic acid and 44.7% docosahexaenoic acid. These optimized conditions were used to study esterification from a PUFA concentrate of the microalgae Phaeodactylum tricornutum and Porphyridium cruentum. With the first, a triglyceride yield of 96.5%, without monoglycerides and very few diglycerides, was obtained after 72 h of reaction; the resulting triglycerides had 42.5% eicosapentaenoic acid. A triglyceride yield of 89.3% was obtained from a P. cruentum PUFA concentrate at 96 h of reaction, which contained 43.4% arachidonic acid and 45.6% EPA. These high triglyceride yields were also achieved when the esterification reaction was scaled up 5-fold.     

Effects of UV-B irradiated algae on zooplankton grazing

We tested the effects of UV-B stressed algae on grazing rates of zooplankton. Four algal species (Chlamydomonas reinhardtii, Cryptomonas sp., Scenedesmus obliquus and Microcystis aeruginosa) were used as food and fed to three zooplankton species (Daphnia galeata, Bosmina longirostris and Brachionus calyciflorus), representing different taxonomic groups. The phytoplankton species were cultured under PAR conditions, and under PAR supplemented with UV-B radiation at two intensities (0.3 W m^–2 and 0.7 W m^–2, 6 hours per day). Ingestion and incorporation experiments were performed at two food levels (0.1 and 1.0 mg C l^–1) using radiotracer techniques. The effect of food concentration on ingestion and incorporation rate was significant for all three zooplankton species, but the effect of UV-B radiation was more complex. The reactions of the zooplankton species to UV-B stressed algae were different. UV-B stressed algae did not affect Daphnia grazing rates. For Bosmina the rates increased when feeding on UV-B stressed Microcystis and decreased when feeding on UV-B stressed Chlamydomonas, compared with non-stressed algae. Brachionus grazing rates were increased when feeding on UV-B stressed Cryptomonas and UV-B stressed Scenedesmus, and decreased when feeding on UV-B stressed Microcystis, compared with non-stressed algae. These results suggest that on a short time scale UV-B radiation may result in increased grazing rates of zooplankton, but also in decreased grazing rates. Long term effects of UV-B radiation on phytoplankton and zooplankton communities are therefore difficult to predict.     

Studies on dissimilatory sulfate-reducing bacteria that decompose fatty acids

Three strains (2ac9, 3ac10 and 4ac11) of oval to rodshaped, Gram negative, nonsporing sulfate-reducing bacteria were isolated from brackish water and marine mud samples with acetate as sole electron donor. All three strains grew in simple defined media supplemented with biotin and 4-aminobenzoic acid as growth factors. Acetate was the only electron donor utilized by strain 2ac9, while the other two strains used in addition ethanol and/or lactate. Sulfate served as electron acceptor and was reduced to H₂S. Complete oxidation of acetate to CO₂ was shown by stoichiometric measurements with strain 2ac9 in batch cultures using sulfate, sulfite or thiosulfate as electron acceptors. With sulfate an average growth yield of 4.8 g cell dry weight was obtained per mol of acetate oxidized; with sulfite or thiosulfate the growth yield on acetate was about twice as high. None of the strains contained desulfoviridin. In strain 2ac9 cytochromes of the b- and c-type were detected. Strain 2ac9 is described as type strain of the new species and genus, Desulfobacter postgatei.     

Studies on dissimilatory sulfate-reducing bacteria that decompose fatty acids

Gliding motility, ultrastructure and nutrition of two newly isolated filamentous sulfate-reducing bacteria, strains 5ac10 and 4be13, were investigated. The filaments were always attached to surfaces. Growth was supported by addition of insoluble aluminium phosphate or agar as substrata for gliding movement. Electron microscopy of ultrathin sections revealed cell walls characteristic of Gramnegative bacteria; the undulated structure of the outer membrane may pertain to the translocation mechanism. Intracytoplasmic membranes were present. Acetate, higher fatty acids, succinate or fumarate served as electron donors and carbon sources. Strain 5ac10 grew also with lactate, but not with benzoate that was used only by strain 4be13. Strain 5ac10 was able to grow slowly on H₂ plus CO₂ or formate in the presence of sulfate without additional organic carbon source. The capacity of complete oxidation was shown by stoichiometric measurements with acetate plus sulfate. Both strains contained b- and c-type cytochromes. Desulfoviridin was detected only in strain 5ac10. The two filamentous gliding sulfate reducers are described as new species of a new genus, Desulfonema limicola and Desulfonema magnum.     

Response of algae and zooplankton to C18 fatty acids of Chlamydomonas reinhardtii

Three C₁₈ fatty acids were assayed for their activity against a number of algae and zooplankton. The three acids, lenolenic, lenoleic, and oleic, reduced the growth of Haematococcus lacustris, Synechococcus leopoliensis, and Botrydiopsis alpina by 50% of control growth in concentrations below 7 ppm. Calanoid and cyclopoid copepods in mixed cultures inoculated with lenolenic and lenoleic acid had LD₅₀ values below 10 ppm. An increase in copepod mortality was observed with increases in cyclopoid density and decreased with increases in calanoid density. Eucyclops agilis inoculated with lenolenic acid had a LD₅₀ value of 4 ppm.     

Biomass productivity and productivity of fatty acids and amino acids of microalgae strains as key characteristics of suitability for biodiesel production

Microalgae are discussed as an alternative source for the production of biofuels. The lipid content compared to cultivation time of used species is the main reason for any choice of a special strain. This paper reviews more analytical data of 38 screened microalgae strains. After the cultivation period, total content of lipids was analysed. The extracted fatty acids were quantified as fatty acid methyl esters by GC analysis. The amino acids were analysed by HPLC. Chlorella sp., Chlorella saccharophila, Chlorella minutissima and Chlorella vulgaris were identified as species with the highest productivity of fatty acids relevant to transesterification reactions. The components were mainly linoleic acid, palmitic acid and oleic acid. To increase productivity of highly saturated fatty acids, cultivation parameters light intensity and temperature were varied. In this manner, the ideal conditions for biodiesel production were defined in this publication.     

A new sensitive tracer for the determination of zooplankton grazing activity

A new tracer compound is presented for determining zooplanktongrazing activity. The gut content in zooplankton is measuredas ß-dimethylsulphoniopropionate (DMSP), which canbe measured even in individual copepods. Species-specific DMSP/Chl-aratios allow applications in, for example, prey selectivitystudies.     

Polyunsaturated fatty acids in zooplankton: variation due to taxonomy and trophic position

1. Food quality has major effects on the transfer of energy and matter in food webs, and essential long‐chained polyunsaturated fatty acids (PUFAs) can affect the quality of phytoplankton as food. In a study of oligotrophic lakes in north‐western Sweden, we investigated the fatty acid composition of four planktonic cladocerans and two calanoid copepods, representing herbivorous and carnivorous species. We also collected seston samples. 2. The proportions of long‐chain PUFAs in the organisms increased with their increasing trophic position. Thus, both their quality as food for other organisms, as well as their requirement for fatty acids (FAs), differed among taxa and depended on their trophic position. 3. We found taxon‐specific differences in the FA composition of zooplankton that were not related to sestonic FA composition. This implies that the variation in zooplankton FA composition is constrained by phylogenetic origin, life history characteristics, or both. 4. The cladoceran taxa contained 12–23% eicosapentaenoic acid (EPA) but only 0.9–2.1% docosahexaenoic acid (DHA) of the total FA content. In contrast, the calanoid copepods contained 7–11% EPA and 14–21% DHA. Thus, our results show that differences in the PUFA content among zooplankton species could have repercussions for both food web structure and function.