Effects of essential fatty acids and N and P-limited algae on the growth rate of tropical cladocerans

SUMMARY 1. In this study, the effects of nutrient (N and P) deficiency and the importance of essential polyunsaturated fatty acids (PUFA) [eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA)] to tropical cladocerans, growth and reproduction were determined in a growth bioassay.
2. The animals were fed N/P-sufficient, N-deficient and P-deficient algae, and also N and P-deficient algae supplemented with fish oil emulsions rich in EPA and DHA.
3. Cladocerans showed different responses to nutrient-deficient algae and also to supplements of fish oil emulsions. Moina micrura was most sensitive to P-deficient alga and, surprisingly, grew better and produced more eggs in N-deficient alga than in N/P sufficient alga. Ceriodaphnia cornuta was less sensitive, growing well in both N and P-deficient algae. This species, however, had a lower clutch size in N-deficient alga. On the other hand, Daphnia gessneri was the most sensitive to mineral limitation, showing decreased growth and clutch size in both nutrient-deficient algae.
4. The PUFA supplements to nutrient-deficient algae increased growth rates only for M. micrura and C. cornuta , suggesting that these fatty acids are important food requirements for these species.     

Essential fatty acid content and the phosphorus to carbon ratio in cultured algae as indicators of food quality for Daphnia

1. A series of experiments examined growth of Daphnia magna on three algal diets ( Rhodomonas minuta , Scenedesmus acutus and Synechococcus sp.) at varying physiological states [nitrogen and phosphorus (P) limitations] to test whether variation in algal fatty acid and/or elemental composition can predict Daphnia growth.
2. These algae differed widely in their essential fatty acid (EFA) composition while phosphorus (P) or nitrogen limitation had only a small influence on their ω3-polyunsaturated fatty acid (PUFA) content.
3. Individually, algal ω3-PUFA content explained 69% of the variation in the normalised growth of D. magna, while algal phosphorus content explained 11% of the variation. Quantitative models for D. magna growth used both algal ω3-PUFA content and algal C : P ratio as food quality indices. Together, algal ω3-PUFA content and C : P ratio explained 70% of the variation in the normalised growth rate of D. magna .
4. Our results indicate that EFA influenced algal food quality much more strongly than P. The EFA and mineral P impacts appear to be independent.     

Effects of algal food quality on fecundity and population growth rates of Daphnia

1. Food quality was at least as important as food quantity for both fecundity and population growth responses of the cladoceran Daphnia pulicaria fed the green alga Ankistrodesmus falcatus grown under N limitation, P limitation, or non-limited condition.
2. The fecundity of D. pulicaria was reduced under conditions of low food quality (low N or low P) compared with that for animals fed control non-limited algae regardless of ration size. The reduced fecundity of D. pulicaria fed P-limited food could be partially alleviated by increasing the ration (hence, compensation), but such was not the case for animals fed N-limited food.
3. Population growth rates of D. pulicaria ( r _max) were significantly reduced under conditions of low-quality food for both N-limited and P-limited algae. Population growth rates were unaffected by ration size, indicating no compensation.     

Is the fatty acid composition of Daphnia galeata determined by the fatty acid composition of the ingested diet?

1. The fatty acid (FA) composition of Daphnia galeata and their algal food was analysed and showed many similarities, however, some significant differences were also found in the relative abundance of the FA C16 : 4ω3 and docosahexaenoic acid (DHA). Their relative abundances were much lower in daphnids than in their algal diet.
2. When daphnids were fed three distinct emulsion particles with DHA : eicosapentaenoic acid (EPA) ratios of c. 0.7, 2 and 4, the final DHA : EPA ratio in the daphnids always favoured EPA. The increase of the food DHA : EPA ratio resulted in a minor increase of DHA (to c. 2%). Feeding the animals on emulsion particles with increasing ratios of DHA : EPA, caused a minor ( c. 2%) increase of DHA level but EPA levels remained high ( c. 10%).
3. When labelled with [¹⁴C]linoleic acid and [¹⁴C]linolenic acid daphnids showed low conversion of both essential FA into C20 polyunsaturated fatty acids (PUFAs). This low conversion activity might explain the importance of C20 PUFAs as dietary compounds in the food of Daphnia.
4. The results indicate the insignificance of DHA and C16 : 4ω3 for daphnids. As EPA can be derived from C18 : 3ω3 it is not strictly essential, although it might be a significant factor in food quality for Daphnia.     

Influence of food quality on growth and reproduction in Daphnia

SUMMARY. 1. Growth and reproduction were measured for Daphnia pulex (Leydig) fed Scenedesmus obliquus (Turp.) Kutz. grown with nitrogen-sufficient and nitrogen-deficient media. The incorporation of carbon into the body of D. pulex was traced using ¹⁴C-labelled algae and biochemical fractionation.
2. Scenedesmus cultured on N-defkient media allocated more carbon to lipid and less to protein than when cultured on N-sufficient media.
3. In a short-term feeding experiment, Daphnia accumulated more lipid when fed N-deficient algal cells high in lipid content than when fed algae grown on N-sufficient media that were lower in lipid.
4. Animals grew faster and produced larger broods when fed algae grown on an N-sufficient medium over an 8-day period. Daphnids fed N-deficient algae had higher quantities of body lipid, suggesting it was not a lack of energy which limited their growth and reproduction.
5. Daphnia fed algae grown on N-deficient media allocated more lipid to eggs, and their offspring lived longer under starvation conditions.     

Particulate fatty acids in two small Siberian reservoirs dominated by different groups of phytoplankton

SUMMARY 1. We studied the composition of fatty acids (FAs) in the seston from two small freshwater reservoirs (Bugach and Lesnoi) with distinct periodicity of domination by cyanobacteria and eukaryotic algae during the growth season.
2. The diatoms in the both reservoirs were characterised by a high content of 14:0 and C16 unsaturated acids, whereas that of the essential FA 20:5ω3 [eicosapentanoic acid (EPA)] was low. The correlation between this polyunsaturated FA (PUFA) and diatom biomass was not significant in either reservoir. The percentage of 20:5ω3 in seston significantly correlated with the biomass of euglenophyta in Bugach and dinophyta in Lesnoi. Hence the diatoms, usually referred as a valuable food for zooplankton, were not an important source of the essential PUFA in these systems.
3. The dominant cyanobacteria in Bugach, and the green algae in Lesnoi, both contained the same marker acids: 18:3ω3 and 18:2ω6. Hence, a discrimination between these two phytoplanktonic groups on the basis of FA biomarkers may be difficult in some cases.
4. We found no significant correlation between the content of 20:5ω3 in seston and the biomass of the dominant daphniids in either reservoir. This is contrary to expectations, based on the literature, that EPA is generally important. Rather, the biomass of the two dominant Daphnia species in Bugach correlated strongly with the content of 18:3ω3 in the seston. The cyanobacteria were a probable source of this ω3 FA for Daphnia . We conclude that EPA is not always important for Daphnia populations although, in such cases, some other PUFA (e.g. 18:3ω3) might be related to their growth.     

Effect of the addition of polyunsaturated fatty acids to the diet on the growth and fecundity ofDaphnia galeata

1. The importance of long-chain polyunsaturated fatty acids (PUFAs), in particular eicosapentaenoic acid (EPA), for the growth and development of Daphnia galeata (Sars) was tested using food types differing in PUFA and EPA contents.
2. Life history experiments of D. galeata fed with the cryptophyceans Rhodomonas lacustris (Pascher & Ruttner) and Cryptomonas pyrenoidifera (Gentler), and the green alga Scenedesmus acutus (Meyen), showed that both cryptophycean species were higher in quality than S. acutus.
3. Since the cryptomonads contained significant amounts of EPA while no EPA could be detected in Scenedesmus , tests were performed to ascertain whether EPA was responsible for the differences in food quality. Feeding daphnids a mixed diet of Scenedesmus and emulsion particles that were rich in EPA and DHA, resulted in a significant improvement in the intrinsic population growth rate. The initial difference in food quality between Scenedesmus and the cryptomonads was completely compensated for by addition of emulsion to the Scenedesmus food.
4. From the observed stimulatory effect of the addition PUFA to the daphnid diet, this study concludes that the presence of such long-chain PUFA improves food quality for daphnids.     

Changes in food quality of the green alga Scenedesmus induced by Daphnia infochemicals: biochemical composition and morphology

1. The effects of Daphnia infochemicals on the morphology and biochemical composition of Scenedesmus were studied and subsequently their influence on Daphnia life history. Three species of Scenedesmus ( S. acutus, S. obliquus and S. subspicatus ) were tested for Daphnia -induced colony formation. Life history experiments were performed with Daphnia cucullata and D. magna feeding on unicellular or colonial S. acutus.
2. Colony formation was promoted when S. acutus and S. obliquus were exposed to filtered water from a D. magna culture. S. subspicatus did not form colonies when exposed to culture water, but showed some colony formation when exposed to live D. magna.
3. No clear differences were found in total lipid, protein or carbohydrate content between the unicellular and colonial ecomorphs of the three Scenedesmus species. However, fatty acid (FA) concentration and composition were changed. Total FA as a percentage of frozen dry weight (DW) were increased in colonies. The ratio of monounsaturated (MUFA) and polyunsaturated fatty acids (PUFA) and the ratio of ω3 : ω6 fatty acids decreased in colonies.
4. A short-term grazing experiment with three size classes of Scenedesmus revealed lower clearance rates for small Daphnia when feeding on large colonial Scenedesmus.
5. Life history parameters of both Daphnia species were influenced by food type. The intrinsic rate of population increase ( r) was significantly lower for daphnids feeding on colonies.
6. The negative influence of colonial Scenedesmus on population growth of Daphnia seems to be determined by algal morphology rather than biochemical composition.     

Effects of food type on the life history of Daphnia clones from lakes differing in trophic state. I. Daphnia galeata feeding on Scenedesmus and Oscillatoria*

1. Effects of the green alga Scenedesmus obliquus and the filamentous cyanobacterium Oscillatoria limnetica as food sources on the growth and reproduction of nine Daphnia galeata clones were studied. A high concentration of these two algae species was fed to Daphnia clones collected from four lakes with varying trophic conditions in a life history experiment.
2. Although Oscillatoria is regarded as a low quality food for Daphnia because of its filamentous morphology and suspected nutritional inadequacy, the cyanobacterium sustained both body growth and reproduction in D. galeata. Growth and reproduction were, however, lower and the onset of reproduction was delayed on a diet of Oscillatoria compared to a diet of Scenedesmus. The intrinsic growth rate, r , for each D. galeata clone was positive on a diet of Oscillatoria , ranging from 0.06 to 0.16 mm day^–1, whereas the intrinsic growth rates on a diet of Scenedesmus ranged from 0.27 to 0.34 mm day^–1.
3. The degree of growth depression on the Oscillatoria diet varied among clones. Clonal responses were correlated with body size; clones with larger body size decreased their growth and reproduction more on a diet of Oscillatoria. The responses of different clones from the same lake to the two diets resembled each other, irrespective of geography or trophic state of the lake.     

Life history consequences for Daphnia pulex feeding on nutrient-limited phytoplankton

1. The growth and feeding of Daphnia pulex De Geer on different algal species was examined. The green algae Chlamydomonas reinhardtii Dangeard and Scenedesmus acutus Meyen, the diatom Synedra tenuissima Kützing, the cryptophyte Cryptomonas pyrenoidifera Geitler and the cyanobacterium Microcystis aeruginosa Kützing were cultured in non-limiting and in N- or P-limiting medium and used as food for D. pulex.
2. Growth limitations were reflected in the elemental and biochemical composition and the morphological characteristics of the algal resources.
3. The clearance rates of D. pulex feeding on nutrient-limited algae were reduced. This was not observed when nutrient-limited mutant Chlamydomonas cells without cell walls were used as food, indicating that the cell wall may play an important part.
4. In comparison with animals grown on nutrient-sufficient cells, nutrient-limited algae resulted in smaller body length, reduced brood sizes, reduced size at maturity, increased age at first reproduction and, consequently, in reduced Daphnia population growth rates.
5. Daphnia population growth rates ( r ) were negatively correlated with the C : P ratio and the carbohydrate content of the food. Moreover, significant correlations between r and clearance rates were found.
6. The observed differences in the grazing and the life history parameters of Daphnia feeding on non-limited and nutrient-limited algae may be the result of both reduced nutritional value and reduced digestibility of nutrient-limited algae.     

The importance of highly unsaturated fatty acids in zooplankton nutrition: evidence from experiments with Daphnia, a cyanobacterium and lipid emulsions

1. We used laboratory growth and feeding experiments to evaluate the role of ω3 highly unsaturated fatty acids (HUFA) in zooplankton nutrition. Polyunsaturated fatty acids (PUFA) comprised 40% of total fatty acids (FA) in the green alga Scenedesmus acutus but only 6% in the cyanobacterium Synechococcus elongatus.Scenedesmus contained modest amounts of the ω3 HUFA eicosapentaenoic acid (EPA, 20 : 5ω3) and docosahexaenoic acid (DHA, 22 : 6ω3), whereas Synechococcus contained only a trace of EPA and no DHA. 2. One-day-old Daphnia maintained high growth rates for 2–3 days on a diet of Synechococcus but exhibited reduced growth or even lost weight over the later part of the 6- or 7-day experiments. Daphnia magna grew better than D. galeata or D. pulicaria on a diet of pure Synechococcus. Daphnids fed the same concentration (0.5 mgC l^–1) of green algae or a mixture of green algae and Synechococcus maintained high growth rates. 3. Supplementing Synechococcus with fish oil emulsions rich in ω3 HUFA markedly improved the growth and reproduction of all three Daphnia species. Supplementation with an emulsion of monounsaturated oleic acid (18 : 1ω9), however, did not affect the growth of D. galeata and caused a decrease in the growth of D. magna. 4. Short-term (7 min) and long-term (3 h) feeding trials with ¹⁴C-labelled Synechococcus were used to evaluate the effects of acclimation and mixed diets on carbon incorporation. D. galeata that had fed on unlabelled Synechococcus for 3 days exhibited no decline in clearance rate but a marked decline in carbon incorporation in comparison with animals acclimated with Scenedesmus or fed a mixture of Synechococcus and Scenedesmus. 5. Our results support the hypothesis that the poor nutritional quality of a cyanobacterium is at least partially due to a HUFA deficiency. Growth and feeding experiments both suggest that the utilization of Synechococcus is enhanced by HUFA from lipid reserves or a mixed diet.     

Food quantity and quality regulation of trophic transfer between primary producers and a keystone grazer (Daphnia) in pelagic freshwater food webs

The transfer of energy and nutrients from plants to animals is a key process in all ecosystems. In lakes, inefficient transfer of primary producer derived energy can result in low animal growth rates, accumulation of nuisance phytoplankton blooms and dissipation of energy from the ecosystem. Most research on carbon transfer efficiency in pelagic food webs has focused on either food quantity or food quality, with the latter considered separately as either elemental stoichiometry or biochemical composition. The natural occurrence and magnitude of these types of growth limitations and their combined effects on Daphnia , a keystone grazer in pelagic freshwater ecosystems, are largely unknown. Our empirical models predict that the strength and nature of food quantity and quality limitation varies greatly with lake trophic state (total phosphorus, TP) and that Daphnia growth rates and thus energy and nutrient transfer efficiency are highest in lakes with intermediate trophic status (TP 10–25 μg l⁻¹). We predict that food availability place the greatest constraint on Daphnia growth in nutrient poor lakes (TP≤4 μg l⁻¹). Phosphorus limitation of Daphnia growth increased with decreasing TP, but the overall effect was never predicted to be the dominant constraining factor. Eicosapentaenoic acid (EPA, 20:5ω3) limitation was predicted to occur in both nutrient poor and nutrient rich lakes and placed the primary constraint on food quality in the most productive lakes. Two contrasting EPA-models gave different results on the magnitude of EPA-limitation, implying that additional food quality factors decrease Daphnia growth at high TP. In conclusion, the model predicts that Daphnia growth should peak in mesotrophic lakes, food quantity will place the greatest constraint on growth in oligotrophic lakes and EPA will primarily limit growth in eutrophic lakes.     

Evidence of phosphorus-limited individual and population growth of Daphnia in a Canadian Shield lake

We performed bag experiments in a Canadian Shield lake with generally high seston (suspended food particles mainly composed of algae) carbon (C):phosphorus (P) ratios, and investigated the responses of individual and population growth of herbivorous Daphnia dentifera on their abundance with (+P) and without (−P) a phosphorus enrichment to lake water. In both treatments, increased abundance of D. dentifera reduced seston C concentration and was accompanied by decreases in population and individual growth rates. However, P-enrichment increased seston P concentration and then reduced seston C:P ratio from 400–700 to ca 100 (by atoms). As a result, both individual and population growth rates were significantly higher in the +P treatment at all animal abundances even though seston C concentrations were similar between the treatments. The magnitude of the growth enhancement by the P-enrichment was independent of animal abundance. Stepwise regression analyses revealed that 71 and 90% of the variance in the population and individual growth rates, respectively, were explained by seston C and P concentrations, and that the contribution of the seston P concentration was roughly the same as that of seston C. Such joint effects of seston C and P indicate that food quality (P content) as well as food quantity (C concentration) can influence Daphnia not only at the level of individual growth but also at the level of population dynamics in P-limited lakes. Our results thus strongly corroborate the hypothesis that the population development of a key herbivore Daphnia in P-limited Canadian Shield lakes is inhibited by the direct effects of P-limited food on individual growth, which weaken the strength of trophic cascading interactions starting from piscivorous fish through planktivorous fish and zooplankton to algae.     

Grazing resistance in nutrient-stressed phytoplankton

Grazing experiments were performed with the zooplankters Daphnia pulex and Daphnia magna, feeding on phosphorus-saturated and phosphorus-limited cells of two green algae (Scenedesmus subspicatus and Selenastrum capricornutum). P-limited algal cells passed largely intact through the gut and were thus spared from heavy grazing pressure. P-saturated algal cells, in contrast, were efficiently assimilated. Structural and morphological changes in the P-limited cells most probably reduced their digestibility. This phenomenon may be an important factor in zooplankton production and competition, and may serve as an example of a highly efficient strategy of P-limited algae to resist heavy grazing pressure.     

Effects of UVB-irradiated algae on life history traits of Daphnia pulex

1. The impact of ultraviolet-B (UVB)-irradiated phytoplankton on the life history parameters of Daphnia was studied. Three species of Chlorophyceae ( Chlamydomonas reinhardtii, Scenedesmus acutus and S. subspicatus ) and one species of Cryptophyceae ( Cryptomonas pyrenoidifera ) were cultured with and without exposure to UVB radiation (biologically effective dose of 3 kJ m^–2 day^–1 DNA weighted) and fed to Daphnia pulex.
2. The daphnids feeding on UVB-irradiated algae tended to be smaller in size, to produce less and smaller offspring, and to have a reduced population growth rate.
3. Fatty acid (FA) analysis of the phytoplankton showed a reduced total FA content under UVB radiation for all four species. Changes in the content of unsaturated fatty acids (UFA) were species specific.
4. The results indicate that the energy transfer between the primary producers' and grazers' level might be negatively influenced by UVB radiation. Direct effects on food quality are modest but detectable, changes in quantity or species composition may be more important in the energy transfer.     

Trophic interactions in rockpool food webs: regulation of zooplankton and phytoplankton by Notonecta and Daphnia

We studied trophic interactions in experimental rockpools with three different food web structures: phytoplankton and small-bodied zooplankton; phytoplankton, small-bodied zooplankton and Daphnia ; and phytoplankton, small-bodied zooplankton, Daphnia and Notonecta . Nutrients, primary productivity, chlorophyll a and zooplankton species composition and biomass were measured over eight weeks.
2. Daphnia had a negative impact on other zooplankton and reduced the phytoplankton biomass and primary productivity. In the absence of Daphnia , small-bodied zooplankton species were abundant, in particular cyclopoid copepods. Concentrations of dissolved nutrients were lower and the standing crop of primary producers was higher when Daphnia was absent.
3. The presence of the invertebrate predator Notonecta produced a top-down effect which was similar to that reported for planktivorous fish, i.e. a selective reduction of daphnids followed by an increase of small-bodied zooplankton species and phytoplankton biomass.
4. The study showed that consumer regulation of Daphnia by Notonecta and of algae by Daphnia are important, but also demonstrated that trophic level biomasses were controlled by a combination of predation and resource limitation.     

Modelling interactions of food quality and quantity in homeostatic consumers

1. Boundaries between growth limitation by carbon (C) and phosphorus (P) in homeostatic heterotrophic consumers such as zooplankton will vary as demands on these two elements vary, as they should at different food quantities. At very low food quantity when production is close to zero, metabolic requirements (for carbon) become more important than growth requirements (for carbon and phosphorus in fixed proportion). Thus, the boundary separating C-and P-limited growth should be at a higher C : P ratio at low food quantity than at high food quantity.
2. A model including both metabolism and growth indicates that consumer growth should differ between foods of high vs. low phosphorus concentration only when food quantity is above a certain level. Thus, two foods might give identical consumer growth rates at low food quantity but give different consumer growth at higher quantity.
3. Solution of the model using parameters based on 2 mm Daphnia compared with a survey of C and P in seston of marine and freshwater sites supports earlier conclusions of the potential importance of food quality constraints on Daphnia growth.     

Effects of natural and commercial diets on the fatty acid content of European grayling

Eicosaenoic acid (20:1ω9) and docosaenoic acid (22:1ω11) levels were about 10 and 100 times higher in food pellets fed to cultured grayling than in the insect larvae on which wild grayling fed. Among the PUFA, docosahexaenoic acid (DHA) was very high in the pellets, resulting in an unnaturally elevated, and probably unbalanced, ω3/ω6 ratio of 7–13 in the cultured fish whereas the same ratio varied only from 4 to 6 in the wild fish. Despite very low DHA levels in the native food, wild grayling muscle tissue contained relatively high amounts of DHA. DHA is probably not essential in the diet of grayling.     

Influence of biochemical,mineral and morphological features of natural food on tropical cladocerans

Five laboratory experiments were performed to evaluate the effect of supplements of fatty acids and a green alga on the individual growth and reproduction of three species of tropical cladocerans Ceriodaphnia richardi, Daphnia ambigua, and D. gessneri feeding on natural seston from the Brazilian Lake Monte Alegre. Cohorts of newborns from cultivated females were submitted to one of the following treatments: (1) Natural seston, (2) Natural seston + microcapsules of EPA and DHA or linoleic and linolenic fatty acids, (3) Natural seston + oil-free microcapsules, and (4) Natural seston + green alga Scenedesmus spinosus (1 mg C l⁻¹). Particulate organic carbon, algal carbon, C:P ratios of seston and green alga, polyunsaturated fatty acids (PUFA) content of seston and cladocerans, as well as phytoplankton composition, size, and shape were measured. The addition of fatty acids to seston did not significantly enhance growth and reproduction of the cladocerans, suggesting that sestonic PUFA content is sufficient for promoting cladoceran development, even in the cool–dry season when the fatty acids used in the experiments were 5–10 times lower in the seston than in the warm–wet season. Despite high C:P molar ratios in most experiments, there was only one indication of growth limitation by P. Reproduction was more affected than individual growth on some occasions by food quantity (energy) caused apparently by algal size, morphology, and digestion resistance. Energy availability, which is affected by algae morphological characteristics, seems to prevail over PUFA and P in controlling growth and reproduction of cladocerans in tropical Lake Monte Alegre.     

Clonal variation in the survival and reproduction of Daphniapulicaria under low-food stress

1. Survivorship and reproductive parameters of eight Daphnia pulicaria clones were evaluated in a life-table experiment under four dietary regimes ranging from food limitation to complete food deprivation.
2. D . pulicaria exhibited both diet-independent interclonal differences and genotype–environment interactions in life-span.
3. Reproduction ceased entirely or nearly so under the three treatments with lowest food availability. Under life-long food limitation, differences between clones were evident in clutch sizes and age at first reproduction. Some clones produced ephippia at a markedly higher frequency than others under food-limiting conditions.
4. For most of the life-history traits evaluated, within-lake variation among clones exceeded differences between the two lake populations they represented.
5. Intraspecific variations in response to periods of food deprivation and to extended quantitative or qualitative food-limitation could affect the clonal composition of Daphnia populations. They are therefore of potential importance in determining the effect of altered phytoplankton assemblages on zooplankton communities.