大型和小型萼花臂尾轮虫的遗传和生活史特征比较研究

近年来，有关生物个体大小变异规律的研究已经成为生活史对策研究的重要内容之一。研究发现，大小相似的萼花臂尾轮虫（Brachionus Calyciflorus）母体所产休眠卵孵化出的不同克隆后代个体大小变化显著，其中，最大个体是最小个体体积的6.25倍。推测种群内产生不同大小的后代个体是轮虫应对环境变化的一种适应性进化策略，然而目前对上述不同大小轮虫克隆的遗传和生活史特征的研究尚未见报道。以mtDNA COI基因和rDNA ITS序列作为分子标记，比较了个体大小差异显著的不同克隆萼花臂尾轮虫的遗传分化程度和分类地位，并在不同温度（20℃、25℃、30℃）和不同斜生栅藻（Scenedesmus obliquus）食物密度（1.0×10⁶、3.0×10⁶、5.0×10⁶个/mL）下比较了它们的生活史特征。结果表明，萼花臂尾轮虫种群内个体大小变异并非由于遗传特征的明显分化所导致，大型和小型个体轮虫克隆在两种分子标记上并不构成姐妹种，且两种形态型间还存在共享单倍型。温度、食物密度、轮虫形态型，以及温度和食物密度各自与轮虫形态型之间的交互作用，均显著影响轮虫的生活史特征。小型轮虫在1.0×10⁶个细胞/mL食物密度下显著延长了胚胎和幼体的发育时间，缩短了生殖期历时；大型轮虫在1.0×10⁶个细胞/mL和3.0×10⁶个细胞/mL食物密度下显著延长了幼体的发育时间，但是其用于胚胎发育和生殖的时间却不随食物密度的变化而变化。各温度和食物密度条件下，大型轮虫的生殖期历时、平均寿命和世代时间均显著延长，或有延长趋势；而两者的种群增长能力之间的差异却因温度和食物密度的不同而异。20℃、25℃以及3.0×10⁶个细胞/mL和5.0×10⁶个细胞/mL食物密度下两种形态型轮虫的生殖能力相似；30℃条件下小型轮虫的生殖能力更强；1.0×10⁶个细胞/mL食物密度下大型轮虫的生殖能力更强。小型轮虫在各温度和各食物密度下均未产生混交雌体后代，而大型轮虫在20℃低温下有较高的后代混交率。因此，大型和小型个体轮虫克隆具有显著不同的生活史策略，且利用有性生殖直接产生个体体积明显变异的不同克隆后代是萼花臂尾轮虫适应不可预测环境变化的一种"赌注策略"。     

在培养基和食料中添加铅和镉对轮虫种群动态的影响

由于工业活动的影响,墨西哥水体环境中的重金属浓度在上升.浮游动物,尤其是轮虫类,由于对环境变化十分敏感而且是淡水中的常见组成部分,因此被广泛用于生态毒理试验以确定水质标准.在不同的胁迫途径下(如通过培养基或食料),重金属的毒性是不同的.在本研究中,通过在轮虫Brachionus rubens的培养基和食料中添加重金属这两种途径,我们评估了镉和铅的效应.对于这两种重金属,均采用将轮虫置于含0.5×106个/ml绿藻的培养基中或每天喂食经5倍于LC50值的金属处理(1,2和4h)的绿藻.对于在培养基中添加镉,使用了三个毒性水平(0.1,0.2和0.4 mg/L),铅的浓度分别为0.005,0.010 和0.015 mg/L.基于LC50的数据,B.rubens对铅的敏感性要比镉高24倍.镉浓度为0.4 mg/L时,培养基中加入镉造成B.rubens的生长趋缓.而喂食经不同时间处理的绿藻后,轮虫的密度随着食料在重金属中处理时间的延长而减小.培养基中或食料中添加铅时,轮虫种群生长的的趋势与在镉处理下的情况类似.随着培养基中重金属浓度的增加,每天种群增长率(r值)会减小.在培养基和食料处理两种不同途径下,r值会在0.33(对照)到0.02 d-1(经重金属处理)间变化[动物学报 51(1)46-52,2005].     

Experimental study on the impact of dinoflagellate Alexandrium species on populations of the rotifer Brachionus plicatilis

To investigate harmful effects of the dinoflagellate Alexandrium species on microzooplankton, the rotifer Brachionus plicatilis was chosen as an assay species, and tested with 10 strains of Alexandrium including one known non-PSP-producer (Alexandrium tamarense, AT-6). HPLC analysis confirmed the PSP-content of the various strains: Alexandrium lusitanicum, Alexandrium minutum and Alexandrium tamarense (ATHK, AT5-1, AT5-3, ATCI02, ATCI03) used in the experiment were PSP-producers. No PSP toxins were detected in the strains Alexandrium sp1, Alexandrium sp2.Exposing rotifer populations to the densities of 2000 cells ml⁻¹ of each of these 10 Alexandrium strains revealed that the (non-PSP) A. tamarense (AT-6) and two other PSP-producing algae: A. lusitanicum, A. minutum, did not appear to adversely impact rotifer populations. Rotifers exposed to these three strains were able to maintain their population numbers, and in some cases, increase them. Although some increases in rotifer population growth following exposures to these three algal species were noted, the rate was less than for the non-exposed control rotifer groups.In contrast, the remaining seven algal strains (A. tamarense ATHK, AT5-1, AT5-3, ATCI02, ATCI03; also Alexandrium sp1 and Alexandrium sp2) all have adverse effects on the rotifers. Dosing rotifers with respective algal cell densities of 2000 cells ml⁻¹ each, for Alexandrium sp1, Alexandrium sp2, and A. tamarense strains ATHK and ATCI03 showed mean lethal time (LT₅₀) on rotifer populations of 21, 28, 29, and 36h, respectively. The remaining three species (A. tamarense strains AT5-1, AT5-3, ATCI02) caused respective mean rotifer LT₅₀s of 56, 56, and 71 h, compared to 160 h for the unexposed “starved control” rotifers. Experiments to determine ingestion rates for the rotifers, based on changes in their Chlorophyll a content, showed that the rotifers could feed on A. lusitanicum, A. minutum and A. tamarense strain AT-6, but could graze to little or no extent upon algal cells of the other seven strains. The effects on rotifers exposed to different cell densities, fractions, and growth phases of A. tamarense algal culture were respectively compared. It was found that only the whole algal cells had lethal effects, with strongest impact being shown by the early exponential growth phase of A. tamarense. The results indicate that some toxic mechanism(s), other than PSP and present in whole algal cells, might be responsible for the adverse effects on the exposed rotifers.     

Herbivore consumers face different challenges along opposite sides of the stoichiometric knife‐edge

Anthropogenic activities have reshaped the relative supply rates of essential elements to organisms. Recent studies suggested that consumer performance is strongly reduced by food that is either very high or very low in relative phosphorus content. However, the generality of such ‘stoichiometric knife‐edge’ and its underlying mechanisms are poorly understood. We studied the response of a planktonic rotifer to a 10‐fold food carbon : phosphorus (C : P) gradient and confirmed the existence of the stoichiometric knife‐edge. Interestingly, we observed a complete homeostatic breakdown associated with strong growth reductions at high food C : P. In contrast, at low food C : P, animals maintained homeostasis despite pronounced performance reductions. Our results suggest that the mechanisms underlying adverse effects of stoichiometric imbalance are determined by both the identity of elements that are limiting and those that are present in excess. Negative effects of excess P reveal an additional way of how eutrophication may affect consumers.     

Sibling species and cryptic speciation in the Brachionus plicatilis species complex (Rotifera)

An attempt is made to identify sibling species within the Brachionus plicatilis complex (Rotifera). Allozyme and morphological data for nine strains from all over the world are provided. Although the analysis of morphological data classified individuals from nine strains into two groups, cluster analysis using genetic distance data obtained from allozyme data revealed at least three groups. A male choice design is described for the first time in rotifers and was used to test for male mating preferences among sympatric strains belonging to three distinct species. The results suggest that the B. plicatilis complex is actually composed of more than three sibling species.     

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.     

pH值对萼花臂尾轮虫种群增长及繁殖的影响

采用种群积累培养法,实验观察了 ｐＨ在３．５～１１．５之间（间隔 １）萼花臂尾轮虫 （Ｂｒａｃｈｉｏｎｕｓ ｃａｌｙｃｉｆｌｏｒｕｓ）种群的增长及繁殖．结果表明,该轮虫种群在ＰＨ６．５～８．５之间增 长较快, ８． ５时增长最快,即瞬时增长率 ｒ和种群密度较大和最大; ｐＨ在 ３． ５～４． ５和 ９． ５ ～１０． ５之间,种群为负增长,即 ｒ为负值; ｐＨ在 ５． ５～９． ５之间种群为正增长,即正为正 值．该轮虫存活的ｐＨ上限为１１．５,下限为３．５．在种群增长最适ｐＨ（８．５）条件下,该轮虫 的繁殖最快,即绝对带卵量最高（１３２个·ｍｌ－１）;ｐＨ在９．５时,其相对带卵量最高．为其它 ｐＨ值条件下的２～４倍．本研究结果可为淡水轮虫的大批量培养提供可靠的ｐＨ值技术指 标．     

Protein and lipid content of the rotifer Brachionus plicatilis during variable growth and feeding condition

Rotifers (Brachionus plicatilis) grown atdifferent growth rate ( = 0.05–0.39 d^–1)were analyzed for protein, lipid, fatty acids, aminoacids and free amino acids, and values are expressedin terms of individuals and dry weight. Increase ingrowth rate is equivalent with increased food rationof the individual rotifer, which responded by higheregg ratio. The protein content per individualrotifer increased by 60–80% when the growth rate increased, whereas the protein content per dryweight showed a slight, although insignificant,increase (p > 0.05). The lipid content perindividual was constant, whereas lipid per dryweight decreased when the growth rate increased. Theratio DHA/EPA decreased when the growth ratesincreased. The amino acids profile in percent oftotal amino acids showed low variation betweencultures maintained at different growth rates,whereas the values expressed in terms of amino acidper individual showed higher variation. The range ofvariation for free amino acids was more pronouncedthan for total amino acids.Short-term food enrichment of poorly fed rotifers( = 0.05 d^–1) with balanced protein richdiet resulted in increased protein and lipid contentper rotifer. The protein content per dry weightshowed only minor changes whereas lipid per dryweight increased. Contrary, short term enrichmentwith a lipid rich diet resulted in increased lipidcontent per individual rotifer and per dry weight,whereas the protein content per individual remainedconstant and the protein content per dry weightshowed a slight decrease.Our experiments show that the amount of protein, wasquite variable in rotifers, and that feeding andgrowth condition were decisive factors affecting it.The range of variation was large enough to be animportant factor during first feeding of marinelarvae, and should therefore be considered infeeding larvae.     

Application of unicellular algae Chlorella vulgaris for the mass-culture of marine rotifer Brachionus

Condensed suspension of Chlorellavulgaris was used for the food of the rotifer Brachionus plicatilis and B. rotundiformis inplace of Nannochloropsis oculata. Thisreport describes the characteristics of C. vulgaris as arotifer food in comparison with N. oculata and thepresent status of this field.The cell components of C. vulgarissuch as protein content, amino acids, minerals andvitamins are generally similar to those of N. oculata. However, the taxonomic status of thesealgal species are different. Based on thesimilarity of cell components, the dietary value ofC. vulgaris is equal in value to that of N. oculata for rotifer growth. Dietary value ofC. vulgaris can be improved by addition ofvitamin B₁₂. This improved C. vulgaris is currently widely used as an indispensable food organism for rotifer culture. Recent investigationshave shown that the use of the condensed suspensionof C. vulgaris makes it possible tosignificantly increase the rotifer density atharvest. Application of condensed C. vulgaris has made rotifer culture quite easy because theculture of N. oculata is no longer required,and intensive rotifer production in aquaculture cannow be realized.     

Temperature conditions enhancing resting egg production of the euryhaline rotifer Brachionus plicatilis O. F. Müller (Kamiura strain)

We examined the effect of low temperature treatment(12°C), followed by transfer to highertemperature (25°C), on resting egg formation ofthe rotifer Brachionus plicatilis, Kamiurastrain. This strain has been mass cultured as livefeed at Kamiura Station (Japan Sea FarmingAssociation) for 9 years at 20°C without theappearance of sexual reproductive stages.After preculture in 20 l of 27 seawater at 12°C for 0, 10, 20, and 30 days,rotifers were inoculated into 0.5 l mass cultures andcultured at 25°C for 7–9 days. The inoculationdensities were changed from 20 to 400 ind. ml^–1,depending on mixis rate. Condensed and frozen Nannochloropsis oculata was fed to rotifers at thefeeding rate of 0.14 µg (dry weight)rotifer^–1day^–1. The control was cultured at12°C for the entire 36 day experiment. No mixisappeared and no resting eggs were produced when thelow temperature treatment was 0 or 10 days. However,mixis rates reached 50-60% after 20 or 30 days ofexposure to 12°C. The number of resting eggsproduced in these treatments reached 25,500 about 13 times higher than the control. Our resultssuggest that low temperature stimulated mictic femaleproduction and the transfer to the high temperatureaccelerated resting egg formation. This method may beuseful for producing resting eggs of rotifer strainsthat lack sexual reproduction in the common culturecondition at larval rearing facilities.