Population growth dynamics of the rotifer Brachionus plicatilis cultured in non-limiting food condition

Population growth parameters in batch culture of Brachionus plicatilis under a continuous supply of freeze-dried microalgae powder have been determined. Two B. plicatilis strains (L- and S-types) and four microalgae species (Nannochloropsis oculata, Nannochloropsis gaditana, Nannochloris oculata and Tetraselmis suecica) have been tested, establishing the dynamics of growth at different daily food rations. Cultures showed a short lag phase, an exponential growth phase, a long post-exponential growth phase and long decline with episodic increases. In both rotifer strains, the best growth was obtained with Nannochloropsis oculata and the poorest with Nannochloris oculata.     

Use of swimming speed and egg ratio as predictors of the status of rotifer cultures in aquaculture

This study evaluated the use of egg ratio (eggs rotifer^–1) and swimming speed (mm min^–1) as prediction criteria for production and culture quality in mass cultures of the rotifer Brachionus plicatilis. Egg ratio was determined to be a suitable predictor of rotifer growth and production in the cultures. Low egg ratios (i.e., 0–0.17 eggs rotifer^–1) indicate reduced rotifer population over time (i.e., negative net population growth rates). However, at this time egg ratio dynamics are not suitably understood to predict in advance a sudden population collapse.Swimming speed of reproductive, egg-carrying females in the exponential growth phase was 40–45 mm min^–1. During exponential growth swimming speed was independent of the food used. Lower swimming speeds were obtained in late stationary phase (10–25 mm min^–1) when yeast was used as a food source. Both environmental factors (e.g., accumulating metabolites) and changes in nutritional state of the rotifers may have affected the swimming speed, but environmental factors appear to be the most important. We believe that swimming speed has the potential of becoming an accurate predictor of culture quality in mass cultures of rotifers.     

Interactions between red tide microalgae and herbivorous zooplankton: effects of two bloom-forming species on the rotifer <Emphasis Type="Italic">Brachionus plicatilis</Emphasis> (O.F. Muller)

Experiments were carried out to investigate interspecific interactions between the rotifer Brachionus plicatilis and two harmful algal bloom (HAB) species using single and mixed culture methods. B. plicatilis populations and the growth of two algae were compared at different algal cell densities. The results demonstrate that B. plicatilis obtained sufficient nutrition from Alexandrium tamarense to support net population increase. When exposed to a density of 8 × 10⁴ cells ml⁻¹ A. tamarense, the number of B. plicatilis increased faster than it did when exposed to other four algal densities (16 × 10⁴, 24 × 10⁴, 32 × 10⁴, and 40 × 10⁴ cells ml⁻¹). Cell densities of A. tamarense decreased due to the grazing of B. plicatilis. In contrast, Heterosigma akashiwo had an adverse effect on the B. plicatilis population and its growth was largely unaffected by rotifer grazing. In this case, the B. plicatilis population decreased and H. akashiwo grew at a rate similar to that of a control without addition of rotifers. Mixed culture experiments showed that A. tamarense could partly counteract the effect of H. akashiwo in limiting the rate of population increase of rotifer. In addition, the effect of different initial cell densities on interspecific competition between A. tamarense and H. akashiwo in mixed culture(s) was also investigated. The results show that A. tamarense competed very successfully when the inoculation proportions of A. tamarense and H. akashiwo were 40:5 and 40:30. Handling editor: D. Hamilton     

Life history characteristics of Brachionus plicatilis (rotifera) fed different algae

A detailed study of the life history of the rotifer Brachionus plicatilis was done at 20 °C, 20 ppt salinity, and 90 mg C 1^–1 food concentration. Rotifers were grown individually in culture plate wells (150 µl culture volume) and fed Isochrysis galbana Tahiti, Tetraselmis sp., Nannochloris atomus, or a l : 1 mixture (weight) of two of the algae. Observations were made every 2–8 hr and rotifers were sized and transferred to new food daily. A total of 19 different parameters were compared. Rotifers fed Isochrysis averaged 21 offspring per female, a 6.7 day reproductive period, a lifespan of 10.5 days and a mean length of 234 µm. After Isochrysis, the foods giving the highest growth, survival, and reproduction in decreasing order were Isochrysis + Nannochloris, Nannochloris, Isochrysis + Tetraselmis, Tetraselmis + Nannochloris, and Tetraselmis. Although the small volume culture system used in this study seems appropriate for studying life history of B. plicatilis, the results cannot always be directly applied to larger cultures.     

Atkinsiella infection in the rotiferBrachionus plicatilis

Atkinsiella infection in the rotiferBrachionus plicatilis has not hitherto been reported. The first known case of this infection is described herein. The fungal growth in vitro was inhibited by 0.05µg/ml of malachite green or 15.6 ppm of formalin. It was experimentally demonstrated that the fungus is pathogenic to the swimming crab,Portunus trituberculatus in the zoeal stage.     

Production and nutritional quality of the rotifer Brachionus plicatilis fed marine Chlorella sp. at different cell densities

The effect of different cell densities of marine Chlorella sp. on the growth rate, doubling time and production of the rotifer Brachionus plicatilis was investigated. A significant increase in rotifer production was achieved at a density of 50 × 10⁶ Chlorella cells ml^–1. The nutritional quality of rotifers grown at different concentrations of Chlorella is discussed.     

Feeding kinetics of Brachionus plicatilis fed Isochrysis galbana

Clearance and ingestion rates of Brachionus plicatilis were measured using ¹⁴C-labeled Isochrysis galbana Tahiti. Experiments were conducted at 20–22 °C, 20 ppt salinity, and algal concentrations ranging from 0.13–64 mg C 1^–1. Clearance rates were constant and maximal at concentrations <2 mg C 1^–1, with maximum rates ranging from 3.4–6.9 µl ind.^–1 hr^–1. The ingestion rate varied with food concentration, and was described by a rectilinear model. The maximum ingestion rate varied considerably, and was dependent on the growth rate of the rotifers. Depending on the pre-conditions, B. plicatilis ingested about 0.5 to 2 times its body carbon per day at saturating food concentrations.     

Studies on life history characteristics of <Emphasis Type="Italic">Brachionus plicatilis</Emphasis> O. F. Müller (Rotifera) in relation to temperature,salinity and food algae

Effects of temperature (18, 24, and 30°C), salinity (5–40 ppt, five intervals) and algal foods (Synechococcus sp., Chlorella pyrenoidosa, Isochrysis zhanjiangensis, Dunaliella salina and Tetraselmis cordiformis) on the life table demography of six geographical Brachionus plicatilis sensu stricto clones, which had been identified according to the prevalent taxonomy and biometric analysis of B. plicatilis sensu lato, were studied. The results showed that temperature, salinity and temperature × salinity significantly influenced the life history parameters. Genotype (clone) had no effect on the population growth rate but did influence the net reproductive rate, generation time and lifespan. All rotifer clones showed the expected increase in population growth rate with increasing temperature. B. plicatilis s. s. attained a higher population growth rate at low–medium salinities (5–20 ppt) than at high salinities (25–40 ppt). The equivalent spherical diameter (ESD) of food algae, salinity and ESD × salinity had significant effects on the life history parameters. In this case, genotype had no effect on population growth rate, net reproductive rate and generation time but did influence lifespan. The population growth rate of B. plicatilis s. s. evaluated against particle retention spectrum of algae at two salinities resulted in bell-shaped curves. Dunaliella salina with an ESD = 7.7 μm was considered to be the best food for B. plicatilis s. s. while Synechococcus appeared to be an inadequate food algae.     

Response of Tetraselmis suecica to nutrient and grazer manipulation

Three methods of algal quantification (direct cell counts, chlorophyll a extraction, in vivo fluorescence) were used to evaluate the response of the unicellular green flagellate Tetraselmis suecica to nutrients and grazers. Nutrient enrichment enhanced total cell counts, chlorophyll a concentration and in vivo and DCMU-fluorescence. Photosynthetic efficiency was reduced in the complete F2 medium as indicated by the high level of in vivo fluorescence, whereas photosynthetic efficiency was increased by the introduction of mussels to the F2 medium. The addition of mussels significantly increased the proportion of non-motile cells, but did not reduce the total cell count. The effect of mussel grazing on algae could be underestimated if only total cells were counted or only the chlorophyll a concentration was measured. The results indicate that these three methods measure different properties of an algal culture and are complementary to each other in assessing the quality and quantity of an algal population. Direct algal counting offers a reliable numerical assessment for cell population abundance. Chlorophyll a concentration was closely correlated to the total cell count. In the presence of mussels, in vivo fluorescence did not correlate with either algal cell counts or chlorophyll a concentration, indicating that the measurement of in vivo fluorescence may be misleading for estimating algal abundance under different culture conditions. This revised version was published online in August 2006 with corrections to the Cover Date.     

Effects of diet on population development of the rotiferBrachionus plicatilis in culture

Experiments were conducted in order to observe the effect of five diets on the population development of the rotiferBrachionus plicatilis Müller under laboratory conditions. Diets were based on baker’s yeast (Saccharomyces cerevisiae) and the algaeTetraselmis suecica andIsochrysis galbana, mixed, or as simple diets. Growth rates, fecundity and biometric parameters were studied for 15 days. The cultures were divided in a logarithmic phase and a harvesting phase. Rotifers fed onTetraselmis, alone or mixed with yeast orIsochrysis, gave good performances with the best results in all the parameters studied. Average growth rates in all diets were similar during the exponential phase, with values ranging from 0.72 (Tetraselmis andTetraselmis + yeast) to 0.47 (yeast). During the harvesting phase there were high differences between diets, with rates highly reduced in the yeast-group (0.17) and good rates whenTetraselmis was ingested (0.65–0.51). This alga had a positive influence on the rotifers, increasing individual growth and fecundity.     

Continuous culture of the rotifer Brachionus plicatilis fed recycled algal diets

A culture system for the rotifer Brachionus plicatilis was designed to maintain higher food conversion rates and stable population densities. Two 2001 plastic tanks were employed in the culture experiments, tank A for feedback culture and tank B for a control culture. The experiments were carried out for 70 days at 24 °C, light intensity, 1500 lux, and a photoperiod of L:D 15:9. B. plicatilis were fed once a day on baker's yeast and Chlorella.Food conversion rates in tanks A and B were 24.7% and 10.1%, respectively. Population density of B. plicatilis in tank A was consistently stable at 100–150 ind. ml^–1 throughout the culture period. Density in tank B, however, showed large fluctuations after 40 or 50 days and by the end of the experiment, declined to zero.     

Microalgal growth enhancement by levoglucosan isolated from the green seaweed <Emphasis Type="Italic">Monostroma nitidum</Emphasis>

Microalgal growth was enhanced by the addition of levoglucosan to the culture medium. The growth-enhancing compound levoglucosan was isolated from the green seaweed Monostroma nitidum using water extraction, molecular fractionation, DEAE-cellulose column chromatography, and high-performance liquid chromatography. Yield of the compound from seaweed powder was 5 × 10⁻³% (w/w). At 10 mM concentration, levoglucosan enhanced cell growth and the specific growth rate of all feed microalgal species tested (Chaetoceros gracilis, Chlorella ellipsoidea, Dunaliella salina, Isochrysis galbana, Nannochloris oculata, Navicula incerta, Pavlova lutheri, Tetraselmis suecica) in most culture media by approximately 150%. Cellular fatty acid profiles and cell size differed marginally between cultures with and without levoglucosan.     

Growth of dominant zooplankton species feeding on plankton microflora in Lake Shira

Batch cultures and continuous flow cultures were used to study the growth rates of zooplankton species from Shira lake, the rotifer Brachionus plicatilis Muller and calanoid copepod Arctodiaptomus salinus Daday, which were fed on phytoplankton and bacterioplankton from the lake. Analyses of the birth and survival rates were used to demonstrate that the lake phytoplankton, consisting mostly of cyanobacteria and diatomaceous algae, is inadequotes for optimal realisation of the reproductive potential of B. plicatilis when compared with the bacterial diet. The study revealed that the kinetic growth characteristics of the two zooplankters were similar: B. plicatilis r _max, 0.120 d^–1; S ₀, 0.253; and K _s, 0.114 mg dry mass l^–1; and for A. salinus r _max, 0.129 d^–1; S ₀, 0.240; and K _s, 0.171 mg dry mass l^–1. Fluctuations in natural food concentration reduced the growth rate of both species. Even though the threshold concentration of food for B. plicatilis and A. salinus were quite similar, the copepods were less sensitive to food limitation.     

Enriching Rotifers with “Premium” Microalgae. Nannochloropsis gaditana

The nutritive quality of Nannochloropsis gaditana cultured semicontinuously with different daily renewal rates was tested as a diet for short-term enrichment of the rotifer Brachionus plicatilis. After 24 h, dramatic differences in the survival, dry weight, and biochemical composition of the rotifers depending on the renewal rate of microalgal cultures were observed. Survival after the feeding period increased with increasing renewal rates. Rotifers fed microalgae from low renewal rate, nutrient-deficient cultures showed low dry weight and organic contents very similar to those of the initial rotifers that were starved for 12 h before the start of the feeding period. On the contrary, rotifers fed nutrient-sufficient microalgal cells underwent up to twofold increases of dry weight and protein, lipid, and carbohydrate contents with regard to rotifers fed nutrient-depleted N. gaditana. Consequently, feed conversion rate decreased in these conditions, indicating a better assimilation of the microalgal biomass obtained at high renewal rates. No single microalgal biochemical parameter among those studied can explain the response of the filter feeder. Similarly to gross composition, EPA and n-3 contents in rotifers fed microalgae from nutrient-sufficient cultures were double than the contents found in rotifers fed nutrient-limited microalgae. In addition, very high positive correlations between the contents of EPA and n-3 in N. gaditana and B. plicatilis were observed. These results demonstrate that selecting the appropriate conditions of semicontinuous culture can strongly enhance the nutritional value of microalgae that is reflected in the growth and biochemical composition of the filter-feeder even in short exposure periods.     

Nitrogen flow through a Brachionus/Chlorella mass culture system

A model of nitrogen flow is presented through the Brachionus plicatilis/Chlorella saccharophila mass (batch) culture system, from the initial input of inorganic nitrogen to the algal culture medium to the final production of rotifers, dissolved nitrogen and particulate nitrogen.A nitrogen budget was first formulated for B. plicatilis relating ingestion, excretion, egestion, somatic growth and reproductive growth. Measurements were made on rotifers from 20° and 10° cultures.The calculated model of nitrogen flow through the rotifer/algal batch culture system estimates the percentages of the original input nitrogen which will be incorporated into algal nitrogen, rotifer nitrogen and the particulate and dissolved nitrogen pools. It is suggested that the dissolved nitrogen pool could be recycled directly for use in subsequent algal culture.     

Atkinsiella parasitica sp. nov. isolated from a rotifer,Brachionus plicatilis

A new species ofAtkinsiella (Lagenidiales, Oomycetes),Atkinsiella parasitica is described and illustrated. It was isolated from the eggs and bodies of a rotifer,Brachionus plicatilis, with fungal infection in 1992. The rotifer was reared in a hatchery as the first food supply for seed production of crustaceans and fishes. The fungus is characterized by producing monoplanetic, lateral biflagellate zoospores, and infrequently branched discharge tubes. Optimum temperature for the fungus was 25°C. The fungus was considered an obligate marine fungus, because its growth was observed only on PYGS medium including seawater.     

High yield mixotrophic cultures of the marine microalga Tetraselmis suecica (Kylin) Butcher (Prasinophyceae)

The effects of three organic compounds were tested on one of the most used marine micro-algae in the aquaculture of molluscs and crustaceans, Tetraselmis suecica. Studies were made in axenic conditions with yeast extract, peptone and glucose added to the culture medium, each alone, in combinations of two or all together. Medium without any organic compound was used for the control. Cultures containing yeast extract grew best, reaching maximum cell density of 3.79 × 10⁶ and 3.84 × 10⁶ cells ml⁻¹. The organic carbon source affected the biochemical composition. The components most affected were the carbohydrates, with values between 6.5 pg cell⁻¹ in control cultures and 48.5 pg cell⁻¹ in glucose cultures. Protein content ranged between 27.5 pg cell⁻¹ in control cultures and 88.6 pg cell⁻¹ in yeast + glucose + peptone cultures. The lipid content changed little. Maximum protein yields were reached in cultures with yeast + glucose and with yeast - glucose - peptone, with values of 24.6 and 28.2 mg 1⁻¹ d⁻¹, respectively. These values are 22 and 25 times those in control cultures. A maximum carbohydrate yield of 7.9 mg carbohydrate per litre per day was obtained in yeast + glucose + peptone cultures, 27 times that in the control cultures. The maximum lipid yield was obtained with yeast + glucose + peptone and yeast + glucose. Maximum energy values were 308 kcal 1⁻ in yeast extract - glucose - peptone cultures and 279 kcal 1⁻¹ in yeast extract + glucose cultures. Gross energy values in control cultures were 24.5 kcal 1⁻¹, but peptone cultures presented the minimum energy value, 22 kcal 1⁻¹. The yeast extract: glucose ratio in the culture medium was optimized. A ratio 2:1 produced the best yields in cells, protein, carbohydrate and gross energy.     

Combined Effects of Food Concentration and Temperature on Competition Among Four Species of <Emphasis Type="Italic">Brachionus </Emphasis>(Rotifera)

We evaluated the effect of algal food density (1.5 × 10⁶, 3.0 × 10⁶ and 4.5 × 10⁶ cells ml⁻¹ of Chlorella) and temperature (22° and 28 °C) on competition among the rotifers Brachionus calyciflorus, Brachionus havanaensis, Brachionus patulus and Brachionus rubens, based on population growth experiments for 24 days. The growth experiments were conducted seperately for each individual rotifer species (i.e., controls), and in mixtures of all four species in equal initial proportions (i.e., under competition). The population growth of B. calyciflorus, B. havanaensis, B. patulus and B. rubens grown separately at two temperatures and at three algal food densities showed typical patterns of lag, exponential and retardation phases in the controls. This pattern differed considerably under competition. In general, we observed that in all of the test species, the highest growth rates were observed at higher food levels and in the absence of congenerics. At 22 °C, under the lowest food level, the differences in the population abundances of B. havanaensis, B. patulus and B. rubens grown alone and in the presence of competition were large. However, these differences reduced as food density was increased from 0.5 × 10⁶ to 4.5 × 10⁶ cells ml⁻¹. At 28 °C and at the lowest food level, all of the other rotifer species eliminated B. havanaensis in mixed cultures. Each brachionid species had a higher rate when grown alone than when cultured with other species. The highest r (mean ± standard error: 0.54 ± 0.01 day⁻¹) was recorded for B. havanaensis at 28 °C under 4.5 × 10⁶ cells ml⁻¹ of algal food density. At 28 °C at low algal food density, the presence of competitors resulted in negative population growth rates for three of the four rotifer species tested.     

Characterization of the bacterial flora of mass cultivated Brachionus plicatilis

Bacterial density and composition in association of mass cultivated rotifers (Brachionus plicatilis, SINTEF-strain) was investigated, during experimental conditions identical to the procedures used for preparing rotifers as live food for marine cold water fish larvae. These procedures include cultivation, enrichment with squid meal and acclimation to low temperature by storage of the rotifer culture at 6 °C. Large variations were observed in the number of rotifer associated (1.8–7.6 · 10³ colony forming units per rotifer^–1) and free-living (0.6–25 10⁷ cells·ml^–1) bacteria. An increase of 50–150% in the bacterial number was normally observed after feeding the rotifer with squid meal, but after three days of acclimation at 6 °C, the bacterial numbers decreased to the initial level.After enrichment of the cultures with squid meal, the similarity in the composition of the bacterial flora between the rotifers and water was reduced. However, acclimation of the culture at 6 °C resulted in better agreement of the rotifer associated flora and that in water. Enrichment of the cultures induced a shift in the bacterial composition from Cytophaga/Flavobacterium dominance to Pseudomonas/Alcaligenes dominance. The bacterial flora of the rotifer cultures are dominated by presumably opportunistic species after enrichment, which may have detrimental effects when rotifers are fed as live food to marine fish larvae.     

Use of freeze-dried microalgae for rearing gilthead seabream, Sparus aurata L., larvae. II. Biochemical composition

The objectives of this study were to test whether freeze-dried microalgae are nutritionally adequate for rearing rotifers as food for gilthead seabream larvae. The elemental composition (C, N, H) and fatty acid composition were analysed in larvae of gilthead seabream, Sparus aurata L., rotifers Brachionus plicatilis and Brachionus rotundiformis and freeze-dried microalgae Nannochloropsis oculata. Four larval feeding treatments were tested: (A) larvae fed rotifers cultivated with freeze-dried microalgae and daily addition of freeze-dried microalgae to the larval tanks; (B) larvae fed rotifers cultivated with freeze-dried microalgae and daily addition of live microalgae to the larval tanks; (C) larvae fed rotifers cultivated with freeze-dried microalgae, without addition of microalgae to the larval tank and (D) larvae fed rotifers cultivated with live microalgae and daily addition of live microalgae to the larval tanks. No significant differences were observed between the biochemical composition of larvae with treatment A (with freeze-dried microalgae) and the composition of larvae in treatment D that were obtained with the acceptable methods for culture systems (with live microalgae).