Use of the rotifer, Brachionus calyciflorus Pallas, in freshwater ornamental fish larviculture

The Brachionus calyciflorus used in this study were produced by batchculture using Chlorella spp. as feed. Larviculture experiments in indoor10-l and 200-l tanks revealed that, compared with egg yolk, the rotifersused as starter food significantly improved the growth and survival of DwarfGourami larvae (Day 2–12). These beneficial effects also extended tothe subsequent Artemia feeding phase (Day 13–32), suggesting that thequality of starter food is crucial to later development. At metamorphosis,the overall survival rate of larvae fed on rotifers in indoor tanks(65.1–74.5%) was about four times of that obtained in extensiveculture in open ponds (17.5%). In Discus, larvae are dependent on thebody slime of their parent as a nutrient during the first two weeks ofexogenous feeding. Our observation demonstrated that Brown Discus larvaecould be raised in the absence of the parent fish by using rotifers asstarter food followed by Artemia nauplii. Their growth and survival ratewere comparable to those on parental feeding. The artificial feeding wouldeliminate the risk of larvae being eaten by the parent fish and shorten thebrooding interval of the spawners, thereby leading to higher yield of fry.This feeding protocol is less tedious and more practical for use incommercial farming of Discus than the existing strategies of smuggling thebatch of larvae to foster parents or feeding the larvae with egg food. Theuse of rotifers would enable freshwater larviculture to improve larvalperformance, increase yield, and facilitate breeding of new fish specieswith small larvae.     

萼花臂尾轮虫有性生殖、种群增长和休眠卵产量间的关系

利用种群指数增长和Logistic增长模型,通过计算机模拟研究了萼花臂尾轮虫有性生殖发生的频率,后代中的混交雌体发率对种群增长和休眠卵产量的影响。在所模拟的参数范围内,随有性生殖发生频率由100％减小到20％,获得了大休眠卵产量所需的混交雌体百分率由9％增大到69％,随密度制约作用的增大（环境容纳量K值由1000减小到100）,该混交雌体百分率由18％增大到69％,休眠卵产量由1072．10降低至133．67,种群的内禀增长率与获得最大休眠卵产量所需的混交雌体百分率间呈曲线相关。当有性生殖发生得越频繁,种群增长所受的密度制约作用较小时,后代中10％－30％的个体为混交雌体时的种群中休眠卵产量较大。     

食物种类对萼花臂尾轮虫种群增长率的影响

在短期慢性观测过程中,食物类型可能是造成萼花臂尾轮虫(Brachionuscalyciflorus)种群繁殖率变化的一种原因。共观测了分别单独投喂10种不同绿藻对轮虫种群增长率的影响。为验证藻青菌是藻类饵料(如绿藻Scenedesmus)有价值的佐剂这一假说,还用蓝细菌单独投喂或与斜生栅藻(Scenedesmusobliquus)混合投喂轮虫进行实验观测。结果发现食物种类对萼花臂尾轮虫种群增长率影响显著。斜生栅藻组获得最大种群增长(1.6/d),而Desmodesmus组增长率最低(0.3/d)。以占斜生栅藻组最大增长的百分率来表示,其它几种绿藻组种群增长由高到低依次为:Desmodesmussubspicatus88%,小球藻(Chlorellavulgaris)83%,单壳缝藻(Monoraphidiumminutum)77%,D.quadricauda74%,S.falcatus71%,S.acuminatus69%,S.pectinatus64%,莱茵衣藻(Chlamydomonasreinhardtii)57%,D.abundans19%。轮虫增长率的差异不能用藻类饵料的大小差异来解释。蓝细菌(Microcystisaeruginosa)和(Synechococcuselongates)不论是单独投喂还是与优良藻类饵料(斜生栅藻)混合投喂都对萼花臂尾轮虫种群增长有抑制作用。这种副作用似乎与微囊藻素无关。该结果不支持无毒蓝细菌可作为与其他绿藻饵料配合使用的优良佐剂这一假说。本研究所观察到的生长变化显示了饵料种类对萼花臂尾轮虫种群增长的影响,也预示了对毒性实验结果的影响     

High density culture of the freshwater rotifer,Brachionus calyciflorus

The freshwater rotifer, Brachionus calyciflorus is one of the live food organisms used for the mass production of larval fish. In this study possibility of obtaining high density cultures of the freshwater rotifer B. calyciflorus were investigated. The two culture systems used differed in their air and dissolved oxygen supplies using three temperatures in each case: 24, 28 and 32 °C. Rotifers were batch-cultured using 5 l-vessels and fed with the freshwater Chlorella. The growth rate of rotifers significantly increased with an increase in temperature. The maximum density of the rotifers with air-supply at 24 °C, 6500 ind. ml^–1, was significantly lower than those cultured at 28 and 32 °C, i.e. 8600 and 8100 ind. ml^–1, respectively. Dissolved oxygen levels decreased with time and ranged from 0.8 to 1.4 mg l^–1 when the density of freshwater rotifer was the highest at each temperature. The highest density (19200 ind. ml^–1) of freshwater rotifer was obtained in cultures with a supply of oxygen at 28 °C. Densities of 13500 and 17200 ind. ml^–1 were found at 24 and 32 °C, respectively. Levels of NH₃-N increased with time and a dramatic increase of NH₃-N was observed at high temperatures. Levels of NH₃-N at 24, 28 and 32 °C were 13.2, 18.5 and 24.5 mg l^–1, respectively. These levels coincided with the highest rotifer density at each of the three temperatures. When rotifers were cultured with an oxygen-supply and pH was adjusted to 7, the maximum density of rotifer reached 33500 ind. ml^–1 at 32 °C . These results suggested that high density culture of freshwater rotifer, B. calyciflorus could be achieved under optimal conditions with DO value of exceeding 5 mg l^–1 and NH₃-N values of lower than 12.0 mg l^–1.     

Impact of turbulence and turbidity on the grazing rate of the rotifer Brachionus calyciflorus (Pallas)

The impact of turbulence and turbidity on Brachionus calyciflorus grazing rate was determined in short feeding periods (10 min), using labelled Chlorella pyrenoïdosa. The response to water motion of B. calyciflorus depends on it physiological state: the grazing rate of recently fed amictic females stomach green (with one or two eggs) is significantly reduced in agitated environments compared with non-agitated environments. In contrast, the grazing rate of starved amictic females is not reduced by water motion, whatever its velocity (V1=0.18 m s?1 and V2=0.22 m s?1). In the presence of suspended particles (3–6-μm silica beads), a larger reduction in grazing rate is observed in agitated water at any water velocity (V1=0.18 m s?1 or V2=0.22 m s?1), than in stagnant water. A synergy between turbulence and turbidity is unfavourable to feeding of rotifers.     

溴氰菊酯对萼花臂尾轮虫实验种群动态的影响

采用生命表技术对暴露于不同浓度溴氰菊酯溶液中的萼花臂尾轮虫的存活和繁殖进行了研究。结果显示, 溴氰菊酯使轮虫的存活时间显著缩短, 繁殖率降低; 当溴氰菊酯浓度高达3 6 mg/L时, 轮虫的存活时间最短、繁殖率最低。与对照组相比, 除1 2 mg/L外, 其它各浓度的溴氰菊酯均使轮虫的生殖前期显著延长; 浓度为0 6和1 2 mg/L的溴氰菊酯使轮虫的生殖期显著延长, 而浓度为2 4和3 0 mg/L的溴氰菊酯却使轮虫的生殖期显著缩短; 1 2 mg/L的溴氰菊酯使轮虫的平均寿命显著延长。轮虫的生命期望、世代时间、净生殖率和种群内禀增长率均随溴氰菊酯浓度的升高而下降。当溴氰菊酯浓度升高达1 2 mg/L时, 轮虫的净生殖率开始与对照组有显著差异; 而轮虫的种群内禀增长率从溴氰菊酯浓度升高达2 4 mg/L时才开始与对照组有显著差异。在溴氰菊酯的毒性监测中, 净生殖率是比种群内禀增长率更敏感的指标。     

斜生栅藻密度对新安江萼花臂尾轮虫种群动态的影响

以浓度分别为1.0×106、2.0×106、4.0×106和8.0×106cells·mL-1的斜生栅藻(Scenedesmus obliquus)为轮虫食物,在25℃下,应用群体累积培养和单个体培养法研究了食物浓度对采自新安江水域(屯溪段)的萼花臂尾轮虫的种群动态影响。结果表明:食物浓度对轮虫的种群增长率和休眠卵产量均有显著的影响,均表现为低食物浓度下(≤4.0×106cells·mL-1)萼花臂尾轮虫的种群增长率和休眠卵产量较小且无差异,高食物浓度下(8.0×106cells·mL-1)轮虫的种群增长率和休眠卵产量较大;食物浓度对轮虫的混交雌体百分率和受精率无显著影响;在食物浓度为4.0×106cells·mL-1时萼花臂尾轮虫的净生殖率最大,世代时间最长,而其内禀增长率在食物浓度为1.0×106cells·mL-1时最小;在食物浓度为8.0×106cells·mL-1时平均寿命和生命期望最长。     

Does excess carbon affect respiration of the rotifer Brachionus calyciflorus Pallas?

1. Herbivorous zooplankton maintain a rather constant elemental composition in their body mass as compared with the variability commonly encountered in their food. Furthermore, their high phosphorus (P) and nitrogen (N) content means that they often face an excess of carbon (C) in their diet. Regulation of this surplus of energy may occur via modulation of assimilation efficiency, or postassimilation by increased respiration (CO₂) and/or excretion dissolved organic carbon, DOC. Whereas several studies have examined the effect of elemental imbalance in the genus Daphnia, few have examined other zooplankton taxa. 2. We investigated whether the rotifer Brachionus calyciflorus uses increased respiration as a means of stoichiometrically regulating excess dietary C. Growth rate and respiration were measured under different food qualities (C : N and C : P ratios). 3. Both C : N and C : P ratios in food had strong effects on growth rate, demonstrating strong nutrient limitation of rotifer growth when nutrient elements were depleted in the diet and indicating the need for stoichiometric regulation of excess ingested C. 4. Respiration measurements, supported by a stoichiometric model, indicated that excess C was not released as CO₂ in B. calyciflorus and that nutrient balance must therefore be maintained by other means such as excretion of DOC or egestion in faecal material.     

孤雌生殖累积世代数和雌体年龄对萼花臂尾轮虫繁殖的影响

采用单个体培养方法,研究了孤雌生殖的累积世代数和雌体年龄对萼花臂尾轮虫混交雌体形成和产卵量的影响,结果表明,随着轮虫孤雌生殖累积世代数和增加,各代中的总混交雌体百分率呈减小的趋势,年轮的雌体可产生较多的混交雌体,非混交雌体所产后代中的总混交雌体百分率具有随其祖母年龄的增大而增大的趋势,孤雌生殖的累积世代数对轮对轮虫非混交雌体的平均产卵量无显著的影响,非混交雌体的年龄对其报代的平均产卵量亦无显著的影响。     

Ecotoxicological studies with the freshwater rotifer Brachionus calyciflorus

The use of the freshwater rotifer Brachionus calyciflorus as a test organism for ecotoxicological studies was examined. Changes in the feeding-, swimming behavior- and demographic characteristics of the rotifers exposed to increasing concentrations of copper (Cu) were studied and the relationships between the different test parameters and their ecological relevance evaluated.Relatively simple short-term bioassay methods were developed to measure the swimming activity (2 hours) and the filtration (5 hours) rate. Life-table experiments were performed to asses the long-term effects of Cu on the rotifers. Significant reductions in the filtration rate, the swimming activity and the demographic parameter r were observed at 20, 12 and 5 g 1^–1 of Cu, respectively. The implications and the possible ecotoxicological applications of the results are discussed.     

基于rDNA ITS序列分析莲塘湖萼花臂尾轮虫种群遗传结构的季节变化

对采自芜湖市莲塘湖水体中的萼花臂尾轮虫(Brachionus calyciflorus)夏季种群中的30个克隆(S1-S30)和秋季种群中的20个克隆(A1-A20)的rDNA ITS序列进行了分析;以十指臂尾轮虫B.patulus为外群,构建了50个克隆的NJ、MP和ME系统发生树。结果表明:三个系统树均支持将50个克隆分为两个支系;第一支系包括29个夏季采集的单元型(单元型S9除外)和1个秋季采集的单元型A2,第二个支系包括18个秋季采集的单元型(单元型A2除外)和1个夏季采集的单元型S9;两支系间ITS序列差异为5.1%-8.8%;它们应属于两个不同的姐妹种。夏季,莲塘湖水体中的萼花臂尾轮虫种复合体内以第一支系为主构成的姐妹种在相对丰度上占绝对优势,达29/30;而秋季则相反。上述结果表明:浅水湖泊中萼花臂尾轮虫种群密度的季节变动实际上主要在于各姐妹种相对丰度的季节变化;两姐妹种的种群遗传结构季节变化符合重叠模型。     

Genetic differentiation and phylogeographical structure of the Brachionus calyciflorus complex in eastern China

Spatio-temporal patterns and processes of genetic differentiation in passively dispersing zooplankton are drawing much attention from both ecologists and evolutionary biologists. Two opposite phylogeographical scenarios have already been demonstrated in rotifers, which consist of high levels of genetic differentiation among populations even on small geographical scales on the one hand and the traditionally known cosmopolitanism that is associated with high levels of gene flow and long-distance dispersal via diapausing stages on the other hand. Here, we analysed the population genetic structure and the phylogeography of the Brachionus calyciflorus species complex in eastern China. By screening a total of 318 individuals from ten locations along a 2320-km gradient and analysing samples from two growing seasons, we aimed at focusing on both small- and large-scale patterns. We identified eight cryptic species and verified species status of two of these by sexual reproduction tests. Samples in summer and winter yielded different cryptic species. The distribution patterns of these genetically distinct cryptic species were diverse across eastern China, from full cosmopolitanism to local endemism. The two most abundant cryptic species BcWIII and BcSW showed a pattern of strong genetic differentiation among populations and no significant isolation by distance. Long-distance colonization, secondary contact and recent range expansion are probably responsible for the indistinct pattern of isolation by distance. Our results suggest that geographical distance is more important than temporal segregation across seasons in explaining population differentiation and the occurrence of cryptic species. We explain the current phylogeographical structure in the B. calyciflorus species complex by a combination of recent population expansion, restricted gene flow, priority effects and long-distance colonization.     

Near-coronal fluid flow patterns and food cell manipulation in the rotifer Brachionus calyciflorus

Brachionus calyciflorus is able to differentially capture or deflect potential food cells depending on the overall density (numbers µl^–1) and size of suspended particles the rotifer encounters while feeding. This has been shown to correlate with behavioral regulation of pseudotrochal structures, and primarily by large medial tufts of compound cilia (cirri). When pseudotrochal cirri are extended laterally, inferred fluid flow pathlines show that only cells in the central 25 µm of the coronal disk are subject to ingestion. When pseudotrochal cirri form medially-oriented screens, fluid flow is directed away from the pseudotrochas and central corona, an effect apparent up to 70 µm in front of a restrained animal. Fluid flow in all areas of the B. calyciflorus corona is at very low Reynolds Number (Re), with calculated values always less than 10^–1. Accordingly, removal of cells from feeding currents probably is not due to sieving but is more likely due to direct interception of particles by individual ciliary elements. Certain large cells — such as Euglena gracilis — are retained in the inner pseudotrochal space for up to 500 msec, ample time for B. calyciflorus to assess the physical and chemical characteristics of this or other potential foods.     

Reproductive success of the rotifer Brachionus calyciflorus feeding on ciliates and flagellates of different trophic modes

SUMMARY 1. The nutritional value of the bacterivorous ciliate Tetrahymena pyriformis and the algivorous ciliate Coleps sp., as well as the heterotrophic flagellate Chilomonas paramecium and the autotrophic flagellate Cryptomonas ovata , were investigated in population growth experiments using the rotifer B. calyciflorus . The two ciliates, both flagellates, which were of similar size, shape and mobility, were each offered as a sole diet and as a supplement to the alga Monoraphidium minutum , known to support reproduction of B. calyciflorus .
2. To further test nutritional differences between the prey organisms, prey selection experiments were conducted in which B. calyciflorus was able to select between the bacterivorous and algivorous ciliate, and between the heterotrophic and autotrophic flagellate.
3. The results demonstrated that both ciliates and the heterotrophic flagellate were not sufficient to support reproduction of B. calyciflorus when offered as a sole diet. They were, however, a good supplement to algal prey (except for the bacterivorous ciliate T. pyriformis ). In the prey selection experiments, B. calyciflorus positively selected for the algivorous Coleps sp. and the autotrophic C. ovata.
4. Overall, ciliates and heterotrophic flagellates may enhance survival of B. calyciflorus , but reproduction of the rotifer is likely to rely on algal prey. Both higher population growth of B. calyciflorus when fed the algivorous Coleps and the autotrophic Cryptomonas, along with their positive selection, give evidence for prey specific differences in nutrition, with algivorous or autotrophic prey species tending to be of higher nutritional value.     

邻苯二甲酸酯类物质对萼花臂尾轮虫种群增长和有性生殖的影响

以萼花臂尾轮虫(Brachionus calyciflourus)为受试动物,以17β-雌二醇(E2)为阳性对照,运用3d种群增长和4d休眠卵产量实验方法研究了不同浓度的邻苯二甲酸二丁酯(DBP)、邻苯二甲酸二异辛酯(DEHP)和邻苯二甲酸丁苄酯(BBP)对轮虫种群增长和有性生殖的影响。结果显示,与空白对照组相比,0.05-5000μg/L的E2和500μg/L的BBP显著提高了轮虫种群增长率,而0.5和5000μg/L的DBP以及0.05和0.5μg/L的BBP使轮虫种群增长率显著降低;0.5-500μg/L的E2、50和500μg/L的DBP、0.005和500μg/L的DEHP和500μg/L的BBP显著提高了轮虫的休眠卵产量,而5000μg/L的DBP和0.05μg/L的BBP却与其相反;5和500μg/L的E2、50和500μg/L的DEHP以及500μg/L的DBP和BBP均使轮虫混交率显著上升,而0.005和0.05μg/L的E2、5000μg/L的DBP和DEHP及0.5μg/L的BBP显著降低了轮虫种群中混交雌体受精率;除5000μg/L的E2显著降低轮虫种群中的混交雌体数/非混交雌体数之外,其它所有处理组均对其无显著性影响;5、500和5000μg/L的E2、5000μg/L的DBP和5-5000μg/L的DEHP均使轮虫种群中的带卵雌体数/不带卵雌体数显著升高,而BBP对其无显著影响。在实验设置浓度范围内,轮虫种群中的混交雌体数/非混交雌体数与E2浓度间、轮虫种群中的混交雌体受精率和种群增长率与DBP浓度间、轮虫休眠卵产量与DEHP浓度间、轮虫混交率与BBP浓度间均具有显著的效应-剂量关系     

食物浓度对角突臂尾轮虫种群增长、个体大小和卵大小的影响

以蛋白核小球藻为食物 ,应用群体累积培养法研究了食物浓度对角突臂尾轮虫种群增长、个体大小和卵大小等的影响 .结果表明 ,食物浓度对角突臂尾轮虫种群增长率、个体大小和卵大小均有极显著影响 .其中 ,轮虫种群增长率与食物浓度间呈曲线相关 ;当食物浓度为 8.2 4 5 3× 10 6cells·ml-1时 ,种群增长率达最大值 0 .6 0 85d-1;在所研究的食物浓度范围内 ,轮虫个体具有随食物浓度的增大而增大的趋势 ,轮虫卵体积在中等食物浓度范围 (6 .0× 10 6～ 9.0× 10 6cells·ml-1)时较大     

Algae and rotifer turbidostats: studies on stability of live feed cultures

A two stage turbidostat was developed according toBoraas & Bennet (1988), but with highly improvedturbidity sensors. The first stage was an algalturbidostat where algal density was regulated byturbidity measurements. Algal density was also heldconstant in the second stage (rotifer production)according to turbidity measurements. Additionally,the growth rates were monitored. The regulation system allowed an effective on-line process control.Initially, the production of rotifers in long-timestudies was variable. However, after furtherimprovements of the turbidity measurement, fluctuations in the rotifer turbidostat decreasedsignificantly.     

Chronic toxicity of methylparathion to the rotifer Brachionus calyciflorus fed on Nannochloris oculata and Chlorella pyrenoidosa

The effect of sublethal levels of methylparathion (0, 1, 3, 5, 7 mg l^–1) on the freshwater rotifer, Brachionus calyciflorus, during their entire life cycle was studied. Rotifers were fed on two species of unicellular algae: Nannochloris oculata and Chlorella pyrenoidosa; both algal concentrations were 5 × 10⁵ cell ml^–1.The parameters used to determine the toxicity of this compound were survival, fecundity, net reproductive rate (R)_o, generation time (T), intrinsic rate of natural increase (r), reproductive value (V _x/V_o) and life expectancy at hatching (eo). All the demographic parameters studied were affected by methyl-parathion exposure on rotifers fed on both species of algae, but the toxic effect was larger when animals were fed on Chlorella pyrenoidosa; in this case, animals showed a decreased in fertility and also a delayed first reproduction. Sublethal methylparathion levels produced a reduction in most of the parameters selected, especially after exposure to 7 mg l^–1, where the animals died before reproducing.     

Nutritional effect of freshwater Chlorella on growth of the rotifer Brachionus plicatilis

Mass production of Brachionus plicatilis is usually accomplished by feeding so-called marine Chlorella (Nannochloropsis oculata) to the rotifers in marine fish hatcheries. If the marine Chlorella are in short supply, baker's yeast is usually used as a supplementary food. Recently, a condensed suspension of freshwater Chlorella (Chlorella vulgaris, k-22) was commercially developed as another supplementary food. We have evaluated the dietary value of this freshwater Chlorella for growth of the rotifer by means of individual and batch cultures. Rotifers cultured with the freshwater Chlorella suspension under almost bacteria-free conditions, showed very suppressed growth. However if the Chlorella was supplemented with vitamin B₁₂ by adding the vitamin solution into the suspension or by culturing the Chlorella in a medium containing vitamin B₁₂, the nutritional value of freshwater Chlorella was greatly improved and almost at the same level as that of marine Chlorella. Condensed Chlorella may therefore be effective as a supplementary food if vitamin B₁₂ is supplied.