Effect of temperature on resting egg formation of the tropical SS-type rotifer Brachionus rotundiformis Tschugunoff

ABSTRACT:   Tropical minute rotifer strains (SS-type) induce mixis at 30–35°C but sexual reproduction and resting egg formation do not proceed well due to rapid environmental change. The present study examined the effect of temperature regulation on rotifer Brachionus rotundiformis (Langkawi strain, SS-type) resting egg formation in small (500 mL in culture volume)- and large-scale (500 L in culture volume) experiments. Rotifers were cultured at 30°C in 15–17 p.p.t. seawater with an initial density of 1 individual (ind.)/mL. After 4 days, when cultures were in exponential growth stage with active mixis induction, the culture temperature of the experimental rotifers was changed to 25°C. Control rotifers were cultured at 30°C throughout the experiment. Fresh or frozen Nannochloropsis oculata and condensed freshwater Chlorella vulgaris were used as the rotifer diets in the small- and large-scale experiments, respectively. Significantly higher resting egg production was observed with the experimental rotifers (30→ 25°C) versus the control rotifers. In the large-scale trial, experimental rotifers produced 2.6 × 10⁶ resting eggs during a 9-day experiment, which was 1.6-fold more than the control rotifers. Moreover, the efficiency of resting egg formation was found to increase by a factor of 1.8. The present study indicates that decreasing culture temperature from 30 to 25°C after active mixis increased resting egg formation in B. rotundiformis (SS-type).     

The enrichment and retention of ascorbic acid in rotifers fed microalgal diets

The enrichment and retention of ascorbic acid (AA) was investigated in rotifers Brachionus plicatilis fed on microalgae ( Nannochloropsis oculata and Isochrysis sp. (T.ISO)) and baker's yeast Saccharomyces cerevisiae . The concentrations of AA of the rotifer diets used in the study differed significantly: 4200 μg g⁻¹ of dry weight in Isochrysis sp. (T.ISO), 2600 μg g⁻¹ in N. oculata and only 77 μg g⁻¹ in S. cerevisiae . Rotifers contained 620 μg AA g⁻¹ prior to the experimental feeding. When subsequently fed for 3 h on microalgae at a ration of 0.13 mg dry microalgae per 10⁶ rotifers rapidly and efficiently increased their content of AA: Isochrysis sp.-fed rotifers contained 1600 μg AA g⁻¹ and N. oculata -fed rotifers contained 1100 μg AA g⁻¹. Concentrations were boosted by a further feeding of a second ration of algae at three times the initial feeding ration; 21 h later, Isochrysis sp.-fed rotifers contained 2500 μg AA g⁻¹ and N. oculata -fed rotifers contained 1700 μg AA g⁻¹. This represented a 180% and 310% increase in the pre-feeding vitamin concentrations in Isochrysis sp. and N. oculata -fed rotifers, respectively. There were no significant changes in AA concentration in rotifers fed a similar ration of yeast throughout the feeding period (520-620 μg AA g⁻¹). Rotifers retained AA during a subsequent 24 h non-feeding period, with no significant changes in the concentrations in any of the rotifer groups. The production of rotifers rich in AA may be particularly valuable for the culture of fish larvae that have a high requirement for the vitamin.     

Fundamental studies on the physiology of rotifers in mass culture—V. Dry Chlorella powder as a food for rotifers

The dietary value of dried, commercial Chlorella was compared with that of living marine Chlorella, and yeast, in relation to growth of the rotifer Brachionus plicatilis raised individually and by batch culture methods.A concentration of 50 μg/ml of dried Chlorella powder is near an optimal density for rotifer growth. The dried material in suspension is less effective for growth than living marine Chlorella, although it is much more effective than a suspension of yeast at the same density (50 μg/ml).In batch culture (12-l glass vessel), the rotifers grew from an initial inoculation of 13.2 individuals/ml to a density of 434 individuals/ml by the 16th day. About 10⁷ rotifers could be removed from one batch culture in five harvests in the 41-day experimental period.The results indicate that dried Chlorella powder is an effective food for the rotifer Brachionus plicatilis.     

维生素B12对圆型臂尾轮虫种群增长、繁殖和个体大小的影响

为了解维生素B 12 对海水圆型臂尾轮虫( Brachionus rotundiformis )种群增长、繁殖和个体大小的影响,采用四种不同浓度的维生素B 12 (0、0.3、0.6、1.2 μg/mL)强化培养轮虫7 d。每天测定轮虫的种群密度和怀卵率,每隔一天测定轮虫背甲长和宽。试验结果显示:0.3 μg/mL维生素B 12 组轮虫的种群增长、怀卵率显著高于其他组( P<0.05);0.3 μg/mL维生素B 12 组轮虫的背甲长和宽均显著小于其他三组( P>0.05),而其他三组差异不显著。试验结论:在本试验条件下,维生素B 12 对圆型臂尾轮虫种群增长、繁殖和个体大小影响的最适浓度为0.3 μg/mL。     

An indoor solution for mass production of the marine rotifer Brachionus plicatilis Müller fed on the marine microalga Tetraselmis suecica butcher

Mass production of the rotifer Brachionus plicatilis Müller is carried out in a set of 50-liter polyethylene bags in which first the microalga Tetraselmis suecica Butcher is cultured and then the rotifer is grown till most of the algae are grazed.This novel procedure based on a monoxenic production technique, has been conceived with a number of aims in view among which the following should be mentioned: shortest time elapsing between the start and the end of the culture (1 week); minimal care of the cultures; light energy saving for indoor invertebrate mass production systems; unified culture vessel for growing algae and rotifers; accuracy and timing of planned rotifer production. Each bag maintained in the best reported conditions gives over 400 rotifers per ml as final production. Rotifer mass production in plastic bags is better achieved when operated in conjunction with a system of continuous monoxenic cultures of microalgae and of rotifers which provide the inocula.     

Improved performance of an intensive rotifer culture system by using a nitrifying inoculum (ABIL)

Abstract A dense nitrifying culture (ABIL) has been examined for its capacity to stimulate rotifer growth in a labscale culture system. The nitrifiers were applied in different ways. When ABIL was added directly to rotifer batch cultures, it gave rise to significantly higher population densities (factor 1.5–2.5 higher, P  < 0.05). The nitrifiers were subsequently examined for their capacity to enhance the start-up of bioreactors, commonly installed in aquaculture rearing tanks. Of the different carrier materials used in these bioreactors, i.e. CaCO₃, gravel and a PVC matrix (Bionet), CaCO₃ gave by far the best results. In a third set of experiments, effectively nitrifying bioreactor systems were connected to rotifer culture tanks and operated over a period of up to 10 days. It was demonstrated that the ABIL inoculated CaCO₃-based bioreactor allowed excellent rotifer growth reaching rotifer densities up to 5500 rotifers per mL. Moreover, a new system in which the ABIL culture was recirculated through hollow fibres was developed and demonstrated to be effective for supporting rotifer growth up to 3500 rotifers per mL. Overall, the use of the dense nitrifying culture either in seed batch cultures, conventional bioreactors or hollow fibre bioreactor systems in support of rotifer cultures was demonstrated to be effective for improving the water quality and the rotifer growth.     

Optimum Feeding Rate of the Rotifer Brachionus plicatilis on the Marine Alga Nannochloropsis sp.

The optimum feeding rate of the rotifer Bruchionus plicutilis was investigated to determine the best conditions for growth of the rotifer, and also in order to maintain good water quality of the culture. Fifty rotifers of a large size strain were cultured individually at five food density levels of Nannoehloropsis sp. (0.5 ± 10⁶, 1.5 ± 10⁶, 3 ± 10⁶, 5 ± 10⁶, and 8 ± 10⁶ cells/ml). At each level, daily survival and offspring production were recorded until the death of the final individual. The data obtained were analyzed by the life table method. The maximum value of three growth indices (the net reproduction rate, mean life expectancy at age 0, and intrinsic rate of increase) was obtdned at the food density of 1.5 ± 10⁶ cells/ml. The ration size was calculated to be 325% (dry weight)/ day, which is equivalent to about 70% of the saturated feeding rate. It was suggested that the feeding rate should be controlled to lower than the saturated value for an efficient mass culture.     

锯缘青蟹幼体饵料的营养强化

用酵母、水球藻、鱼油强化和豆油强化四种不同方式培养轮虫，再分别投喂锯缘青蟹幼体，分析测定轮虫和体的生化组成，结果显示，（1）不同方式培养的轮虫之间以及摄食这些轮虫的锯缘青蟹幼体之间的蛋白质含量都没有显著差异；（2）轮虫的脂类含量和脂肪酸组成与培养方式密切相关，小球藻轮虫的脂类含量最高，20：5n-3(EPA)占总脂肪酸的比例也最高 ，为18.05%，鱼油轮虫则含有最多的22：6n-3(DHA)，占总脂肪酸3.16%，脂类含量仅次于小球藻轮虫；（3）锯缘青蟹幼体的脂类含量和脂肪酸组成受相应饵料营养成分的影响。另外，幼体培育实验也发现，饵料营养成分影响幼体的存活率，结果表明，提高轮虫的EPA和DHA含量，尤其晨DHA含量，将有利于锯缘青蟹幼体的存活和发育。     

Morphometric changes in a strain of the lineage 'Nevada', belonging to the Brachionus plicatilis (Rotifera) complex

The rotifer Brachionus plicatilis is an important component of aquaculture as a larval feed. Its taxonomic status has been recently re-defined as a species complex, consisting of at least 14 new species/lineages. This study deals with the lineage Brachionus 'Nevada', which has been shown to occur in European hatcheries. A strain of B . 'Nevada' was mass cultured using two commonly applied feeding regimes and analysed in terms of its morphometry. A new formula was proposed for the calculation of volume, which can be used as an index of adequacy of rotifers as feed for fish larvae. The results were related to life cycle parameters. The pre-reproductive and reproductive phases were divided into distinct size groups. Differences were also found between the two diets in morphometry and demography. Rotifers of a larger size (yeast-based diet) showed a lower growth rate and a longer reproductive period, lifespan and mean generation time compared with smaller-sized rotifers (Culture Selco^®-based diet). In terms of lorica length, the present study's strain of B . 'Nevada' (238.5 μm) was intermediate between values reported for Brachionus ibericus (193.5 μm) and B. plicatilis sensu stricto (299 μm).     

几种单胞藻对褶皱臂尾轮虫的营养强化研究

利用气相色谱法测定了小球藻、牟氏角毛藻、球等鞭金藻和投喂不同藻类后12h的褶皱臂尾轮虫的脂肪酸含量。试验结果表明,不论是投喂单种藻类还是2种混合藻类,强化后的轮虫其体内的脂肪酸含量均发生了明显变化,其中单链脂肪酸及低度不饱和脂肪酸的含量在投喂前后没有变化,各种脂肪酸的含量界于未强化的轮虫和所投喂的单胞藻饵料含量之间。3种高度不饱和脂肪酸的含量,除了二十二碳六烯酸因在牟氏角毛藻中含量较低,在单独投喂牟氏角毛藻后的轮虫中未检测出外,其余均与其所投喂的单胞藻的脂肪酸一致;轮虫中被检出的3种高度不饱和脂肪酸的含量,在单种藻类试验组中均低于所投喂藻类的含量;在2种藻类混合试验组中界于2种单胞藻含量之间。混合藻类比单种藻类强化轮虫具有更加全面的营养效果。     

Bacterial colonization of winter flounder Pseudopleuronectes americanus fed live feed enriched with three different commercial diets

The proliferation of bacteria in intensive aquaculture systems may be responsible for poor growth and mass mortality of marine fish larvae. Essential fatty acids provided in the diet could protect larvae by modulation of the immune response via arachidonic acid (AA) and eicosapentaenoic acid (EPA). Winter flounder Pseudopleuronectes americanus larvae were fed rotifers Brachionus plicatilis enriched with three commercial diets containing different fatty acid profiles. Bacterial colonization on the gills and skin and in the intestinal lumen was evaluated at the end of the rotifer feeding period (day 26), and growth was surveyed until metamorphosis. At 26 days post hatching, larvae fed rotifers containing the higher AA content and with a higher docosahexaenoic acid (DHA) to EPA ratio showed better growth and the lowest bacterial colonization of the intestinal lumen compared to larvae fed rotifers with the lowest AA and DHA : EPA levels. AA had been selectively incorporated into the polar lipids of larvae fed the rotifers enriched with the three diets. This is the first study in winter flounder larvae to report a link between different commercial rotifer enrichments and bacterial density in intestinal lumen.     

Effect of Rotifer Enrichment on Sunshine Bass Morone chrysops × M. saxatilis Larvae Growth and Survival and Fatty Acid Composition

The effect of enriching rotifer prey with highly unsaturated fatty acids on sunshine bass Morone chrysops × M. saxatilis larval survival and growth from ages 4 to 12 d posthatch was determined. Comparisons were made among larvae fed (1) rotifers cultured with Nannochloropsis paste versus rotifers cultured with Nannochloropsis paste and enriched with Culture Selco 3000; (2) no rotifers versus rotifers cultured with Nannochloropsis paste and enriched with Culture Selco 3000; and (3) rotifers cultured with Nannochloropsis paste and enriched with Culture Selco 3000, rotifers cultured with Nannochloropsis and Pavlova pastes and enriched with Culture Selco 3000, and rotifers cultured with Nannochloropsis paste and enriched with Culture Selco 3000 and Super Selco. The only differences in survival were unfed larvae with practically no survival compared to 55.4% survival for larvae fed rotifers cultured with paste plus Culture Selco 3000. Larvae fed rotifers cultured with paste plus Culture Selco 3000 were longer and had greater condition than those fed rotifers cultured with paste. Additional enrichment with Pavlova sp. or Super Selco had no affect. A canonical analysis of fatty acid contents of diets, rotifers, and fry supported evidence from harvest results. Distances between centroids indicated distinct differences among diets, less distinction among the rotifers, and little difference among fry. Enrichment enhanced growth, but additional enrichment beyond that done during rotifer culture did not increase survival, growth, or condition.     

Production and Nutritional Quality of Two Small-Sized Strains of the Rotifer Brachionus plicatilis1

The rotifer Brachionus glicatilis, strain-S (from Japan) and strain-S1 (Pozuelo 1975) were mass cultured using Chlorella and bakers' yeast. Under the same feed regime, a significant increase (P < 0.001) in rotifer strain-S production (44.19 ± 8.50 individuals/mL/d) was observed compared with strain-S1 (29.64 ± 9.38 individuals/mL/d). The population growth rate was significantly higher (P> 0.001), and doubling time was significantly lower (P> 0.0001) in rotifer strain-S than in strain-S1. There was a significant decline (P> 0.001) in birth rate of strain-S rotifers in the culture system compared with strain-S1, suggesting short longevity and deterioration of the culture system while using strain-S. In general, no significant difference (P< 0.05) in the presence of ω3 polyunsaturated fatty acids (ω3 PUFA) in different strains of rotifers was observed.     

Insulin/insulin-like growth factor-like activity in the aqueous extracts of the rotifer Brachionus plicatilis

Insulin/insulin-like growth factor (IGF)-like signaling plays important roles in the aging processes of various animals. However, little is known about this signaling in rotifers, which have been used as a model animal in aging studies. Here we report that the aqueous extracts of the rotifer Brachionus plicatilis show activity similar to that of insulin/IGFs. Rotifers were cultured under four different feeding regimens (fed, starved for about ten days, or re-fed for 30 and 120 min after starvation), and then their aqueous extracts were added to culture media of rat L6 myoblasts. Treatment with these extracts increased the phosphorylation levels of mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase (ERK) and two Akt substrates of approximately 48 and 60 kDa, and these phosphorylations were diminished when cells were preincubated with specific inhibitors of their upstream kinases (MAPK/ERK kinase and phosphoinositide 3-kinase, respectively). Furthermore, the extracts from fed rotifers provoked higher phosphorylation levels of MAPK/ERK and Akt substrates than the extract from starved rotifers, suggesting that the production of substance(s) with insulin/IGF-like activity is stimulated upon feeding in the rotifer.     

Effects of yeast and algal diets on the growth and biochemical composition of the rotifer Brachionus plicatilis (Müller) in culture

Abstract. Growth rates and fecundity of Brachionus plicatilis fed baker's yeast, a diatom, and three different flagellates, were monitored during a 15-day feeding trial. Identical growing conditions and equivalent food levels for the different diets were maintained during rotifer culture. The highest growth rate and mean fecundity were exhibited by rotifers fed Chlorella saccharophila , followed in decreasing order by those fed Isochrysis galbana (T-iso), Tetraselmis suecica, Saccharomyces cerevisiae and Thalassiosira pseudonana . Significant differences in contents of neutral lipids, polar lipids, sugars, glycogen, protein, ash and corresponding energy reserves in the resultant five groups of rotifers were diet dependent. Total available energy, about 15 kJ/g in rotifers fed S. cerevisiae and I. galbana (T-iso) was significantly different from the 14kJ/g in rotifers fed T. suecica and C saccharophila, and in turn from the 13kJ/g in rotifers fed T. pseudonana .     

Enrichment of Rotifers Brachionus plicatilis with Eicosapentaenoic Acid and Docosahexaenoic Acid Produced by Bacteria

Abstract— Two bacterial strains, rich in either eicosapentaenoic acid [EPA, 20:5(n-3)] ( Shewanella gel-idimarina ACAM 456) or docosahexaenoic acid [DHA, 22:6(n-3)] ( Colwellia psychroeryrhrus ACAM 605) were tested for their ability to enrich rotifers Erachionus plicatilis in these polyunsaturated fatty acids. Rotifers were exposed for 24 h to each bacterial strain and to a mixture of the two strains. They were then harvested and their fatty acid compositions were analysed and compared to those of rotifers that had been either starved or fed yeast Saccharomyces cerevisiae or microalgae Tetraselmis suecica in 2-L glass flasks. Exposure to 1.4 × 10⁹ cells/ml of the EPA-producing bacterium only resulted in rotifer EPA levels increasing from 0.1% to 1.2% of total dry weight (%dw). Similarly, following exposure to 1.0 × 10⁹ cells/mL of the DHA-producing bacterium only, rotifer DHA levels increased from below detection to 0.1% dw. When exposed to a mixture of the two bacterial strains, containing 7.0 × 10⁸ celldml of the EPA producer and 5.0 × 10⁸ cells/mL of the DHA producer, the rotifers'final EPA and DHA levels were 0.5% dw and 0.3% dw respectively. Although feeding strategies need refining, these results show, for the first time, that rotifers can be enriched with DHA from bacteria, and that rotifers can be enriched simultaneously with both DHA and EPA from different bacterial strains.     

Expression and characterization of the periplasmic cobalamin-binding protein of Photobacterium damselae subsp. piscicida

Cobalamin (vitamin B₁₂) is an essential cofactor in a variety of enzymatic reactions and most prokaryotes contain transport systems to import vitamin B₁₂. A gene coding for a periplasmic cobalamin-binding protein of Photobacterium damselae subsp. piscicida was identified by in silico analysis of sequences from a genomic library. The open reading frame was composed of 834 bp encoding a protein of 277 amino acids. The protein showed 61% identity with the vitamin B₁₂-binding protein precursor of P. profundum , 53% identity with the corresponding protein of Vibrio parahaemolyticus and 43% identity with the periplasmic binding protein BtuF of Escherichia coli. The expression of the native protein was investigated in P. damselae subsp. piscicida , but BtuF was weakly expressed under normal conditions. To characterize the BtuF of P. damselae subsp. piscicida , the recombinant protein was expressed with a C-terminal His₆-tag and purified; the molecular weight was estimated to be approximately 30 kDa. The protein does not contain any free thiol group, consistent with the view that the two cysteine residues are involved in a disulphide bond. The purified BtuF binds cyanocobalamin with an affinity constant of 6 ± 2 μ m .     

Channel Catfish,Ictalurus punctatus,Immunity to Saprolegnia sp.

ABSTRACT

The Japanese flounder, Paralichthys olivaceus, is one of the most common finfish cultured in Japan and Korea. Despite the relatively high production of fingerlings, some problems remain, mainly related to the larval feeding and cost of maintaining microalgae and rotifers. In order to determine the effects of different diets on the Japanese flounder larval growth and survival, a series of experiments was carried out related to the size and nutritional value of different live feeds. The larvae culture conditions were at 10 or 20 larvae/L in 50 to 2,000 L tanks, with aeration and with or without “green water,” and a temperature range of 18.5 to 22.5°C. The live foods used were microalgae (Chlorella ellipsoidea and Nannochloris oculata), baker's yeast, experimental n-yeasts, oyster trochophore larvae, three strains of rotifer Brachionus plicatilis (L-type, S-type and U-type) and Artemia nauplii. Variations were detected in size, dry weight, and chemical composition of the three strains of rotifers used. The maximum number of rotifers ingested by flounder larvae increased steadily from 7 individuals, at first feeding (3.13 mm), to 42 individuals at 5.25 mm of total length (6 days after first feeding). There was a relationship between larval total length and size of the rotifers ingested. The effect of rotifer size on larval growth and survival appeared to be limited to the first two days of feeding. Of the diets tested in the growth and survival of larval flounder during 14 days after hatching, rotifers fed on C. ellipsoidea and raised in green-water gave the best results. Rotifers cultured on enriched N. oculata and n-yeasts did not support larval growth and caused higher mortalities. The n-yeasts used as rotifer enrichment appeared to satisfy, partially, the nutritional requirement of 7-day-old flounder larvae, as did n-yeast squid wintering oil the requirements of 14-day-old larvae. From 7-9-days after hatching and throughout the second 14-day period, rotifers and Artemia cultured on N. oculata improved the survival of flounder compared with those fed on rotifers cultured on C. ellipsoidea. Moreover, the larval growth did not vary significantly between both microalgae-rotifer feedings. No clear relation was found between total protein, lipid, amino acids and fatty acids of live feeds with the growth and survival of flounder larvae, although the total lipid was higher in C. ellipsoidea than in N. oculata. The Artemia nauplii San Francisco strain appeared to be more suitable for the growth and survival of flounder larvae, than the Utah strain. The nutritional value of Artemia nauplii (Utah strain) for flounder larvae remained unchanged despite the use of either microalgae as nauplii enrichment.     

Mass Production of Freshwater Rotifer,Brachionus calyciflorus,Under Different Diets and Regimes

ABSTRACT

The production ability of freshwater rotifer, Brachionus calyciflorus, cultured with two algal species, Chlorella vulgaris, and Spirulina platensis, and a baker's yeast, Saccharomyces cerevisiae, were studied using semicontinuous culture method. B. calyciflorus was fed with the above three types of food at five different concentrations (125, 250, 500,750, and 1,000 μg/mL). Among the three different types of diet, maximum production of B. calyciflorus (489.20 ±10.91 individuals/mL; P < 0.05) was obtained with C. vulgaris, followed by S. cerevisiae, and S. platensis. Moreover, in each of the three diets, the maximum rotifer production was obtained at a particular concentration (C. vulgaris, 750 μg/mL; S. cerevisiae, 750 μg/mL; and S. platensis, 500 μg/mL) beyond which the rotifer production decreased. The peak production due to C. vulgaris (489 individuals/mL) was better than S. cerevisiae (321 individuals/mL) when the number of rotifers was considered. The present study indicates that the quantity and quality of food have a significant role on the rotifer production and that C. vulgaris at 750 μg/mL appears suitable to feed to rotifers for maximal production.     

Waste‐grown phototrophic bacterium supports culture of the rotifer,Brachionus rotundiformis

The present and commonly used batch culture system (BCS) as adopted by many small‐scale tropical hatcheries is beset by poor rotifer production and sudden crashes. This study aimed to produce nutritive rotifers and evaluate their performance based on the BCS by using phototrophic bacteria (PB) that can be easily and cheaply cultured from palm oil mill effluent (POME), an agro‐industrial byproduct usually discarded as waste. Brachionus rotundiformis given a sole diet of POME‐grown PB (Rhodovulum sulfidophilum) grew as well as on the commercially produced microalgae, Nannochloropsis. Production, growth rate and fecundity of rotifers fed condensed bacterial cells (bPB) and culture broth of bacteria grown in POME (cPB) were evaluated. The best performance in terms of the stated parameters was obtained for rotifers fed 200 mL of cPB in 3 L of culture water; this media sustained a mean rotifer density of 600–900 individuals mL^?1 after 3–6 days of culture. The biochemical composition of rotifers fed PB was comparable to those fed microalgae, except that the former contained more polyunsaturated fatty acids.