The use of biomarkers in Daphnia magna toxicity testing

We investigated the effect of short-term exposure to cadmium and 2,4-dichlorophenoxy acetic acid on the digestive physiology of Daphnia magna and the consequences for the bioenergetics of the organism. In both cases, ingestion was more drastically reduced compared to digestive enzyme activity. Furthermore a differential shift in catabolism was noted: in general polysaccharidases were less affected than the enzymes responsible for protein and lipid digestion. Comparison of the ‘1 h in vivo fluorescence’ criterion (Janssen & Persoone, 1993) with the ingestion and digestive enzyme activity revealed that this rapid screening assay should be considered as a quantification of ingestion inhibition rather than a methodology assessing digestive enzyme inhibition. This revised version was published online in July 2006 with corrections to the Cover Date.     

The response of European Daphnia magna Straus and Australian Daphnia carinata King to changes in geomagnetic field

This study investigates the effects of lifelong exposure to reversed geomagnetic and zero geomagnetic fields (the latter means absence of geomagnetic field) on the life history of Daphnia carinata King from Australia and Daphnia magna Straus from Europe. Considerable deviation in the geomagnetic field from the usual strength, leads to a decrease in daphnia size and life span. Reduced brood sizes and increased body length of neonates are observed in D. magna exposed to unusual magnetic background. The most apparent effects are induced by zero geomagnetic field in both species of Daphnia. A delay in the first reproduction in zero geomagnetic field is observed only in D. magna. No adaptive maternal effects to reversed geomagnetic field are found in a line of D. magna maintained in these magnetic conditions for eight generations. Integrally, the responses of D. magna to unusual geomagnetic conditions are more extensive than that in D. carinata. We suggest that the mechanism of the effects of geomagnetic field reversal on Daphnia may be related to differences in the pattern of distribution of the particles that have a magnetic moment, or to moving charged organic molecules owing to a change in combined outcome and orientation of the geomagnetic field and Earth's gravitational field. The possibility of modulation of self-oscillating processes with changes in geomagnetic field is also discussed.     

Phylogeography of Daphnia magna in Europe

The phylogeographical structure of the water flea Daphnia magna in Europe was analysed using a 609-bp fragment of the cytochrome oxidase subunit I gene. Only moderate overall genetic divergence was detected within Europe. We detected four genetically and, to a large extent, geographically distinct phylogroups within Europe. Our results suggest that these groups recolonized large parts of Europe around 100,000 BP from different refugia. Overall, the pattern suggests a high degree of provincialism with a patchy occurrence of specific lineages, thus confirming the highly subdivided genetic structure usually observed in freshwater zooplankton populations. Although the region around the Mediterranean Sea was only sampled patchily, we obtained strong indications for the occurrence of more divergent genetic lineages in this region. Comparing our European samples to samples from North America and Japan revealed a higher level of differentiation, reflecting limited intercontinental dispersal.     

Mechanisms of energy allocation to reproduction in the cladoceran Daphnia magna Straus

Understanding rules of resource allocation within individuals is helpful in explaining population dynamics. This is particularly the case under the conditions of resource limitation that are commonly experienced by zooplankton. Here, we evaluate assumptions underlying models of resource allocation in Daphnia and test the predictions of two models of response to starvation.
We also test the predictions of two simple models concerning the mechanisms of egg provisioning in Daphnia magna. Our results: (1) demonstrate that provisioning is discontinuous, occurring over a discrete period of the instar; (2) support the hypothesis that egg production occurs in a serial fashion; (3) show that Daphnia magna responds to starvation by ceasing egg production but that there is an increase in mean size of eggs provisioned during the instar, prior to starvation; and (4) broadly support predictions that the response of Daphnia to starvation is an instantaneous cessation of reproduction but that there is a time lag before death.     

1,8-二羟基蒽醌对大型溞的毒性效应

The study on the acute,sublethal and chronic toxicity of 1,8-dihydroxyanthraquinone (1,8-dihAQ) to Daphnia magna showed that the 48 h LC₅₀ was 0.37 mg稬^-1,and the feeding behavior of Daphnia magna was severely affected by the compound.When exposed to 0.2 mg稬^-1 of 1,8-dihAQ for 5 h,the filtration and ingestion rate of Daphnia magna was inhibited by 97%.Chronic toxicity test results indicated that the reproduction ability decreased dramatically after exposing to sublethal concentration of 1,8-dihAQ.It could be inferred that reproduction parameters and intrinsic rate of natural increase were the sensitive parameters in characterizing sublethal toxicity.The NOEC and LOEC values for reproduction parameters were also given.     

不同种类农药表面活性剂对大型溞的急性毒性

运用评价化学品对水生生物毒性的标准试验方法,比较了39种非离子型、6种阴离子型和3种阳离子型的常用代表性表面活性剂对大型溞的急性毒性.结果表明:3种阳离子型表面活性剂1427、1227及C8-10的急性毒性均为剧毒,其中1427毒性最高,EC50值为0.97×10-2 mg·L-1;非离子型表面活性剂中蓖麻油聚氧乙烯醚、吐温和斯潘系列乳化剂均为低毒,而烷基酚聚氧乙烯醚系列和脂肪醇聚氧乙烯醚系列表面活性剂的毒性稍偏高,而AEO-7和AEO-5的毒性达到高毒水平,EC50值分别为0.82和0.97 mg·L-1,且此类表面活性剂脂溶性越大,对大型溞的毒性越大;大部分阴离子表面活性剂的毒性为中毒,但NNO表现为高毒,EC50值为0.17 mg·L-1.     

Parasite-mediated selection in experimental Daphnia magna populations

Abstract It has been suggested that parasites are a strong selecting force for their hosts and therefore may alter the outcome of competition among host genotypes. We tested the extent to which parasite-mediated selection by different parasite species influenced competition among clones of the cyclic parthenogen Daphnia magna . We monitored clone frequency changes in laboratory microcosm populations consisting of 21 D. magna clones. Parasite treatments (two microsporidians, Glugoides intestinalis and Ordospora colligata ) and a parasite-free control treatment were followed over a nine-month period. A further treatment with the bacterium Pasteuria ramosa failed. We found significant differences in clonal success among the treatments: the two parasite treatments differed from the control treatment and from each other. Additionally, we measured the clone-specific population carrying capacity, competitive ability against tester clones, and reproductive success of infected and uninfected females to test whether they correlate with clonal success in the microcosms. The clone-specific competitive ability was a good predictor of clonal success in the microcosms, but clonal carrying capacity and host reproductive success were not. Our study shows that parasite-mediated selection can strongly alter the outcome of clonal competition. The results suggest that parasites may influence microevolution in Daphnia populations during periods of asexual reproduction.     

有毒和无毒微囊藻对大型溞生长和繁殖的影响

近年来,随着工农业的快速发展和人们生活水平提高,一些城市生活污水和工农业废水流入江河、湖泊等淡水水体中,加剧了水体的富营养化和污染,导致蓝藻过度生长繁殖而形成水华,其中最常见的是微囊藻水华[1,2].     

Thermal avoidance and preference in Daphnia magna

1. Water fleas (Daphnia magna) bred at 23°C were non-responsive to temperatures between 13 and 25°C.

2. At the lower (11°C) and upper limits (30°C) their klinokinetic avoidance behaviour showed a larger intraindividual than interindividual variation.

3. Thermal sensitivity for avoidance responses in D. magna was about 1.5°C.

4. For D. magna bred for one parthenogenetic generation at 14°C heat avoidance temperature was about 8°C lower, and cold avoidance temperature was about 1°C higher than in D. magna from 23°C.

5. In group experiments the animals showed some preference for the acclimation temperature.

6. Cold induced stenothermy and warm induced eurythermy in D. magna were related to the mode of reproduction.

The use of biomarkers in Daphnia magna toxicity testing. IV. Cellular Energy Allocation: a new methodology to assess the energy budget of toxicant-stressed Daphnia populations

The Cellular Energy Allocation (CEA) methodology wasdeveloped as biomarker technique to assess the effectof toxic stress on the energy budget of testorganisms. This short-term assay is based on thebiochemical assessment of changes in the energyreserves (total carbohydrate, protein and lipidcontent) and the energy consumption (electrontransport activity). The CEA methodology was evaluatedusing Daphnia magna juveniles exposed for 96hto sublethal lindane and mercury chlorideconcentrations. The ecological relevance of the CEAassay was assessed by comparing the sub-organismalresponse with population level parameters (obtainedfrom 21 day life table experiments) such as theintrinsic rate of natural increase (r_m) and themean total offspring per female. Two differentmethodologies were used to assess the effect levels:the no (lowest) observed effect level (NOAECs-LOAECs)approach and the regression-based approach. Bothtoxicants caused a significant decrease in the netenergy budget of D. magna, with a LowestObserved (Adverse) Effect Concentration (LOAEC) of0.18 mg/l and 5.6 µg/l for lindane andHgCl₂,respectively. Changes in the lipid content of theorganisms were detected at toxicant concentrationslower than those affecting the total carbohydrate andprotein content. Toxicant specific effects wereobserved on the electron transport activity.Comparison of the CEA results with those of thepopulation level tests revealed that for mercury theCEA based LOAEC was a three times lower than thatbased on r_m and the total brood size(18 µg/l). For lindane the CEA based LOAEC was twotimes lower than the LOAEC based on r_m(0.32 mg/l) but was higher than that based on thetotal number of offspring produced (0.1 mg/l).Using the regression-based approach, EC₁₀ valueswere calculated using three parameter sigmoid orlogistic models. Comparison between the CEA andr_m based EC₁₀ values demonstrates that forboth chemicals similar effect concentrations areobtained: the CEA-based EC₁₀ (0.20 mg/l) forlindane is 1.5 times higher than the r_m-basedEC₁₀ threshold (0.13 mg/l), while for mercury thebiomarker-based EC₁₀ value (9 µg/l) was 1.4times lower than the population-based EC₁₀ value(12.5 µg/l).From these results, we suggest that the short-term CEAassay may be useful for predicting long-term effectsat the population level. The consequences of theobserved effects on the energy budget of the testorganism are discussed in the context of the effectsemerging at the population and community level.     

The use of biomarkers in the antioxidant responses of Daphnia magna to the acute and chronic exposure to no. 20 diesel oil and 2,4-dichlorophenol

Abstract

The effects of no. 20 diesel oil exposure, 2,4-dichlorophenol (2,4-DCP) exposure and combined exposure on the antioxidant defences of Daphnia magna have been studied systematically for the first time. Daphnia magna was exposed for 1 day or 10 days to several concentrations of 0, 0.005, 0.01, 0.05, 0.1, 0.5 or 1.0 mg L^?1 solutions. Antioxidant defences consisting of the levels of reduced glutathione (GSH) and glutathione disulfide (GSSG) and activities of superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR), selenium dependent glutathione peroxidase (Se-GPx) and glutathione S-transferase (GST) of daphnids were determined to evaluate their protective roles and to analyse the occurrence of oxidative stress. The possible antioxidant defence mechanisms are discussed. Furthermore, GST can be a potential biomarker and an early-warning index for the pollutants in waters in that GST responded sensitively to 1 day and 10 days of exposure to diesel oil and 2,4-DCP and 10 days of combined exposure. Crossover comparisons showed an antagonistic action about the no-observed-effect concentration (NOEC) against Daphnia magna, which needs further studies.     

Genetic diversity of Daphnia magna populations enhances resistance to parasites

The diversity-disease hypothesis states that decreased genetic diversity in host populations increases the incidence of diseases caused by pathogens (= monoculture effect) and eventually influences ecosystem functioning. The monoculture effect is well-known from crop studies and may be partially specific to the artificial situation in agriculture. The effect received little attention in animal populations of different diversities. Compared with plants, animals are mobile and exhibiting social interactions. We followed the spread of a microsporidian parasite in semi-natural outdoor Daphnia magna populations of low and high genetic diversity. We used randomly selected, naturally occurring host genotypes. Host populations of low diversity were initially monoclonal, while the host populations of high diversity started with 10 genotypes per replicate. We found that the parasite spread significantly better in host populations of low diversity compared with host populations of high diversity, independent of parasite diversity. The difference was visible over a 3-year period. Host genotypic diversity did not affect host population density. Our experiment demonstrated a monoculture effect in independently replicated semi-natural zooplankton populations, indicating that the monoculture effect may be relevant beyond agriculture.     

In situ estimates of gut evacuation and its dependence on temperature in five cyprinids

Gut evacuation and its dependence on the temperature in bream Abramis brama , silver bream Blicca bjoerkna , roach Rutilus rutilus , gibel Carassius aurtaus gibelio and carp Cyprinus carpio , held in mesh cages in Lake Balaton, was best fitted by an exponential model in 18 of 36 trials, while in 14 and four trials, a linear and a square-root model, respectively, gave a better fit. Adjusted r ² values, however, often did not differ markedly between the three models. The shape parameter, B , was 0.36 for carp and ranged from 0.81 to 1.24 for the other four cyprinids, according to the general evacuation model expanded with the temperature variable, and fitted to whole data sets. Relationships between the food evacuation rates obtained from the exponential models and the temperature were described by exponential functions for bream, silver bream, roach and gibel.     

The effects of parasitism and inbreeding on the competitive ability in Daphnia magna: evidence for synergistic epistasis

Synergistic epistasis for fitness is often assumed in models of how selection acts on the frequency and distribution of deleterious mutations. Evidence for synergistic epistasis would exist if the logarithm of fitness declines more quickly with number of deleterious mutations, than predicted by a linear decline. This can be studied indirectly by quantifying the effect of different levels of inbreeding on fitness. Here, six sets (different genetic backgrounds) of three increasingly inbred Daphnia magna clones were used to assess their relative fitness according to changes in frequency in a competition experiment against a tester clone. A novelty of the mating procedure was that the inbreeding coefficients (F) of the three clones belonging to each set increased in steps of 0.25 independent of the (unknown) inbreeding coefficient of the common ancestor. The equal increase of the inbreeding coefficients is important, because deviations influence the quantification of inbreeding depression, its variance and the detection of epistasis. In a simple mathematical model we show that when working with a partially inbred population inbreeding depression is underestimated, the variance of fitness is increased, and the detection of epistasis more difficult. Further, to examine whether an interaction between inbreeding and parasitism exists, each inbred clone was tested with and without a microsporidium infection (Octosporea bayeri). We found a nonlinear decrease of the logarithm of fitness across the three levels of inbreeding, indicating synergistic epistasis. The interaction term between parasitism and inbreeding was not significant. Our results suggest that deleterious mutations may be purged effectively once the level of inbreeding is high, but that parasitism seems not to influence this effect.     

Sex ratio of the cyclic parthenogen Daphnia magna in a variable environment

The relationship between environmental factors, sex ratio and mating system in Daphnia magna was examined, and the adaptiveness of environmental sex determination over ametic sex determination was explored. Monthly sexual sex ratios (males over total number of males and sexual females) ranged from 0.31 to 1.0, the three-year average equalling 0.61. However, if only the samples collected during the period of frequent sexuality from August to October are included, the sexual sex ratio becomes equal, 0.51. Sexual sex ratios varied real between samples during the same period and the standard errors appeared highest in July ad August. Typical of suck times is some uncertainty in the environment, and the environmental cues can be contradictory. Sex expression in Daphnia appears to be determined by responses to complicated interactions between different environmental factors, which adaptively alter the sex ratio. The longterm sexual sex ratio of Daphnia aproaching the equilibrium 1:1, despite environmental sex %termination, gives support for Fisier's classic theory of equal parental investment in both sexes. An equal sex ratio is advantageous also during periods of small population size because it maximizes the effective population size.     

温度和食物浓度对大型溞Daphnia magna种群动态和两性生殖的影响

在4个温度(15、20、25、30℃)和3个食物浓度(1×104,1×105,5×105 cells ml-1)下研究了大型溞(Daphnia magna)淮河种群的动态和两性生殖,结果表明:(1)在实验初期,大型溞种群密度不断增加,达到最大值后种群密度呈现缓慢下降或趋于稳定的趋势.除15℃下最大瞬时增长率(1.36 d-1)出现在中食物浓度组外,20℃(1.51 d-1)、25℃(1.39 d-1)、30℃(0.69 d-1)下的最大瞬时增长率均出现在高食物浓度组.(2)相同温度下,食物浓度与大型溞最大种群密度间存在显著正相关(p<0.01).(3)相同食物浓度下,随温度升高,大型溞首次产幼溞时间(5～21d)和到达最大种群密度的时间(15～29d)缩短,首次抱卵时的体长(1.75～2.67 mm)减少.(4)在较低的温度和中食物浓度组下,大型溞产生了较多的休眠卵或卵鞍(15℃:(16.7±2.5) ind. 和20℃:(18.3±3.8) ind.);30℃下没有出现休眠卵或卵鞍.除15℃、中食物浓度组外,其它实验组均产生雄体.实验25d后,雄体密度与种群密度呈显著的相关性(30℃:p<0.05;15～25℃:p<0.01).(5)本研究结果暗示大型溞休眠卵的形成受温度、食物浓度和种群密度的共同影响,且较低的温度是大型溞休眠卵形成的主要诱因.     

五氯酚对大型溞的急性亚慢性和慢性毒性

本文采用换水式试验研究了五氨酚溞（ＰＣＰ）对大型（Ｄａｐｈｎｉａｍａｇｎａ）的急性、亚慢性和慢性毒性,稀释水硬度为８０─１００ｍｇ／Ｌ（以ＣａＣＯ３计）。急性和慢性试验均使用小于一日龄的幼溞,试验温度为２５─２６℃,慢性试验进行了２０ｄ。用小于一日龄幼溞进行的亚慢性试验暴露了１９ｄ,而用四日龄幼溞的亚慢性试验则进行了１６ｄ,水温均保持在１９─２０℃。ＰＣＰ对大型溞的２４ｈ和４８ｈＥＣ５０分别是４８９和２４５μｇ／Ｌ。依据第１胎所产幼溞数求得的最低可观察效应浓度（ＬＯＥＣ）和无可观察效应浓度（ＮＯＥＣ）,在慢性试验中分别是１６０和８０μｇ／Ｌ,在１９ｄ亚慢性试验中分别为２００和１００μｇ／Ｌ,二者相近。试验结果表明,第１胎所产幼溞数是敏感的指标。     

Detection of triclosan using 2-domain hemoglobin promoter of Daphnia magna

Triclosan has been widely used as an antimicrobial agent. However, triclosan was found to cause toxicity, including muscle contraction disturbances, carcinogenesis, and endocrine disorders. In addition, it was found to affect central nervous system function adversely and even have ototoxic effects. Conventional methods for detecting such triclosan can be performed easily. However, the conventional detection methods are inadequate in precisely reflecting the impact of toxic substances on stressed organisms. Therefore, a test model for the toxic environment at the molecular level through the organism is needed. From that point of view, Daphnia magna is being used as a ubiquitous model. D. magna has the advantages of easy cultivation, a short lifespan and high reproductive capacity, and high sensitivity to chemicals. Therefore, the protein expression pattern of D. magna that appear in response to chemicals can be utilized as biomarkers for detecting specific chemicals. In this study, we characterized the proteomic response of D. magna following triclosan exposure via two-dimensional (2D) gel electrophoresis. As a result, we confirmed that triclosan exposure completely suppressed D. magna 2-domain hemoglobin protein and evaluated this protein as a biomarker for triclosan detection. We constructed the HeLa cells in which the GFP gene was controlled by D. magna 2-domain hemoglobin promoter, which under normal conditions, expressed GFP, but upon triclosan exposure, suppressed GFP expression. Consequently, we consider that the HeLa cells containing the pBABE-HBF3-GFP plasmid developed in this study can be used as novel biomarkers for triclosan detection.     

Coping with predator stress: interclonal differences in induction of heat-shock proteins in the water flea Daphnia magna

Although predation is a strong selection pressure, little is known about the molecular mechanisms to cope with predator stress. This is crucial to understanding of the mechanisms and constraints involved in the evolution of antipredator traits. We quantified the expression of heat-shock protein 60 (Hsp60), a potential marker for predator stress, in four clones of the water flea Daphnia magna, when exposed to fish kairomones. Expression of Hsp60 induction increased after 6 h and returned to base levels after 24 h of predator stress. This suggests that it is a costly transient mechanism to temporarily cope with novel predator stress, before other defences are induced. We found genetic variation in the fixed levels and in the fish-induced levels of Hsp60, which seemed to be linked to each clone's history of fish predation. Our data suggest that Hsp60 can be considered part of a multiple-trait antipredator defence strategy of Daphnia clones to cope with predator stress.