Application of rotifer Brachionus plicatilis in detecting the toxicity of harmful algae

The toxicity of seven major HAB (harmful algal bloom) species/strains, Prorocentrum donghaiense, Phaeocystis globosa, Prorocentrum micans, Alexandrium tamarense (AT-6, non-PSP producer), Alexandrium lusitanicum, Alexandrum tamarense (ATHK) and Heterosigma akashiwo were studied against rotifer Brachionus plicatilis under laboratory conditions. The results show that P. donghaiense, P. globosa, P. micans, A. tamarense (AT-6), or A. lusitanicum could maintain the individual survival and reproduction, as well as the population increase of the rotifer, but the individual reproduction would decrease when exposed to these five algae at higher densities for nine days; H. akashiwo could decrease the individual survival and reproduction, as well as population increase of the rotifer, which is similar to that of the starvation group, indicating that starvation might be its one lethal factor except for the algal toxins; A. tamarense (ATHK) has strong lethal effect on the rotifer with 48h LC50 at 800 cells/mL. The experiment on ingestion ability indicated by gut pigment change shows that P.donghaiense, P. globosa, P. micans, A. tamarense (AT-6) and A. lusitanicum can be taken by the rotifers as food, but A. tamarense (ATHK) or H. akashiwo can be ingested by the rotifers. The results indicate that all the indexes of individual survival and reproduction, population increase, gut pigment change of the rotifers are good and convenient to be used to reflect the toxicities of HAB species. Therefore, rotifer is suggested as one of the toxicity testing organisms in detecting the toxicity of harmful algae.     

Experimental study on the interspecific interactions between the two bloom-forming algal species and the rotifer Brachionus plicatilis

The interspecific interactions between the rotifer Brachionus plicatilis and two harmful algal blooms (HAB) species were investigated experimentally by single culture method. B. plicatilis population and the growth of the two algae were compared at different algal cell densities. The results demonstrated that the B. plicatilis obtained sufficient nutrition from Prorocentrum donghaiense to support net population increase. With exposure to 2.5×10⁴ cells mL⁻¹ of P. donghaiense, the number of B. plicatilis increased faster than it did when exposed to other four algal densities (5, 10, 15 and 20 ×10⁴ cells mL⁻¹), and the increase rate of B. plicatilis population (r) at this algal density was 0.104 ± 0.015 rd⁻¹. Cell densities of P. donghaiense decreased due to the grazing of B. plicatilis. In contrast, Heterosigma akashiwo had an adverse effect on B. plicatilis population and its growth was largely unaffected by rotifer grazing. In this case, B. plicatilis population decreased and H. akashiwo grew at a rate similar to that of the control.     

Application of rotifer Brachionus plicatilis in detecting the toxicity of harmful algae

The toxicity of seven major HAB (harmful algal bloom) species/strains, Prorocentrum donghaiense, Phaeocystis globosa, Prorocentrum micans, Alexandrium tamarense (AT-6, non-PSP producer), Alexandrium lusitanicum, Alexandrum tamarense (ATHK) and Heterosigma akashiwo were studied against rotifer Brachionus plicatilis under laboratory conditions. The results show that P. donghaiense, P. globosa, P. micans, A. tamarense (AT-6), or A. lusitanicum could maintain the individual survival and reproduction, as well as the population increase of the rotifer, but the individual reproduction would decrease when exposed to these five algae at higher densities for nine days; H. akashiwo could decrease the individual survival and reproduction, as well as population increase of the rotifer, which is similar to that of the starvation group, indicating that starvation might be its one lethal factor except for the algal toxins; A. tamarense (ATHK) has strong lethal effect on the rotifer with 48h LC₅₀ at 800 cells/mL. The experiment on ingestion ability indicated by gut pigment change shows that P. donghaiense, P. globosa, P. micans, A. tamarense (AT-6) and A. lusitanicum can be taken by the rotifers as food, but A. tamarense (ATHK) or H. akashiwo can be ingested by the rotifers. The results indicate that all the indexes of individual survival and reproduction, population increase, gut pigment change of the rotifers are good and convenient to be used to reflect the toxicities of HAB species. Therefore, rotifer is suggested as one of the toxicity testing organisms in detecting the toxicity of harmful algae. Supported by the Knowledge Innovation Program of Chinese Academy of Sciences (No. KZCX2-YW-208), National Natural Science Foundation of China (No. 40821004, U0733006) and National Basic Research Program of China (973 Program, No. 2001CB409700)     

Impact of several harmful algal bloom (HAB) causing species,on life history characteristics of rotifer <Emphasis Type="Italic">Brachionus plicatilis</Emphasis> Müller

In recent years, harmful algal blooms (HABs) have occurred frequently along the coast of China, and have been exhibiting succession from diatom- to dinoflagellate-dominated blooms. To examine the effects of different diatom and dinoflagellate HABs, the life history parameters of rotifers (Brachionus plicatilis Müller) were measured after exposure to different concentrations of HAB species. The HAB species examined included a diatom (Skeletonema costatum) and four dinoflagellates (Prorocentrum donghaiense, Alexandrium catenella, Prorocentrum lima and Karlodinium veneficum). Compared with the control treatment (CT), the diatom S. costatum showed no adverse impacts on rotifers. Exposure to dinoflagellates at densities equivalent to those measured in the field resulted in a reduction in all the life history parameters measured. This included a reduction in: lifetime egg production (CT: 20.34 eggs/ind.) reduced to 10.11, 3.22, 4.17, 7.16 eggs/ind., life span (CT: 394.53 h) reduced to 261.11, 162.90, 203.67, 196 h, net reproductive rate (CT: 19.51/ind.) reduced to 3.01, 1.26, 3.53, 5.96/ind., finite rate of increase (CT: 1.47/d) reduced to 1.16, 1.03, 1.33, 1.38/d, and intrinsic rate of population increase (CT: 0.39/d) reduced to 0.15, 0.03, 0.28, 0.32/d, for the dinoflagellates P. donghaiense, A. catenella, P. lima and K. veneficum, respectively. The results showed that the diatom S. costatum had no detrimental consequences on the reproduction and growth of B. plicatilis, however, the four dinoflagellates tested did show adverse effects. This suggests that dinoflagellate HABs may suppress microzooplankton, resulting in an increase in algal numbers.     

面包酵母悬浮液中添加脂溶性维生素对褶皱臂尾轮虫增殖率的影响

采用批量培育法研究了在面包酵母悬浮液中添加不同浓度的脂溶性维生素对褶皱臂尾轮虫（Ｂｒａｃｂｉｏｎｕｓｐｌｉｃａｔｉｌｉｓ）增殖率的影响。结果表明：面包酵母对褶皱臂尾轮虫的生长繁殖是有营养缺陷的。在面包酵母悬浮液中．当添加维生素Ａ为０．０２～５．００μｇ／ｍｌ时，随着添加浓度的增大，轮虫的种群增殖率Ｒ值也增大。以添加５．００μｇ／ｍｌ为最佳，第４天的Ｒ值达０．７５，是对照组的５．６倍。添加维生素Ｄ０．００５～０．２０μｇ／ｍｌ对轮虫的增殖有一定促进作用，以添加０．０５μｇ／ｍｌ为最佳，第７天的Ｒ值为０．４０７，是对照组的２．７倍。添加维生素Ｅ０．００１～０．０１μｇ／ｍｌ和添加维生素Ｋ０．０１～０．１０μｇ／ｍｌ时，第４天的Ｒ值与对照组无差异。     

Variability of Prorocentrum donghaiense response to allelopathic action from Alexandrium pacificum in laboratory culture

The dinoflagellates Alexandrium pacificum and Prorocentrum donghaiense are two wellknown harmful algal blooms(HABs)-forming species,both were usually found in the same sea areas in form of HABs in China.To date,there is no mechanistic model that can fully explain the occurrence of P.donghaiense blooms with A.pacificum.We found that different strains of P.donghaiense had different intrinsic growth rates of 0.107-0.215/d,and these strains exhibited different responses to the allelopathic action from A.pacificum.Some strains of P.donghaiense could grow well despite some degrees of inhibition in a short period,suggesting the two algal species P.donghaiense and A.pacificum could coexist,even if A.pacificum was allelopathic.Our findings may advance the understanding of phenotypes in P.donghaiense and provide a potential mechanism involved in the coexistence of P.donghaiense and A.pacificum in the same area.     

Ingestion of <Emphasis Type="Italic">Brachionus plicatilis</Emphasis> under different microalgae conditions

The effects of four microalgae,Chlorella vulgaris,Platymonas helgolandicavar,Isochrysis galbana,and Nitzschia closterium on the grazing and filtering rates of the marine rotifer,Brachionus plicatilis,were evaluated under laboratory conditions.The grazing rates in separate cultures of the four microalga were as follows:C.vulgaris > P.helgolandicavar > I.galbana > N.closterium.However,the filtering rates occurred in the following order:P.helgolandicavar > N.closterium > C.vulgaris > I.galbana.A mixed diets ex...     

Abundance, biomass and composition of spring ice algal and phytoplankton communities of the Laptev Sea （Arctic）

Abundance,biomass and composition of the ice algal and phytoplank-ton communities were investigated in the southeastern Laptev Sea in spring 1999.Diatoms dominated the algal communities and pennate diatoms dominated the dia-tom population.12 dominant algal species occurred within sea ice and underlyingwater column,including Fragilariopsis oceanica,F.cylindrus,Nitzschiafrigida,N.promare,Achnanthes taeniata,Nitzschia neofrigida,Naviculapelagica,N.vanhoef fenii,N.septentrionalis,Melosira arctica,Clindrothecaclosterium and Pyrarnimonas sp.The algal abundance of bottom 10 cm sea icevaried between 14.6 and 1562.2×10~4 ceils l~(-1)with an average of 639.0×10~4cells l~(-1),and the algal biomass ranged from 7.89 to 2093.5μg C l~(-1)with an av-erage of 886.9μg C l~(-1),which were generally one order of magnitude higherthan those of sub-bottom ice and two orders of magnitude higher than those ofunderlying surface water.The integrated algal abundance and biomass of lower-most 20 cm ice column were averagely 7.7 and 12.2 times as those of upper 20 mwater column,respectively,suggesting that the ice algae might play an importantrole in maintaining the coastal marine ecosystem before the thawing of sea ice.Icealgae influenced the phytoplankton community of the underlying water column.However,the“seeding”of ice algae for phytoplankton bloom was negligible be-cause of the iow phytoplankton biomass within the underlying water column.     

Identification of dissolved and particulate carbonyl compounds produced by marine harmful algal bloom species

Carbonyl compounds,especially polyunsaturated aldehydes(PUAs),are increasingly reported as teratogenic to grazers and deleterious to phytoplankton.While PUAs have been considered to be mainly produced by diatoms after cell wounding,little is known about whether microalgae other than diatoms,particularly harmful algal bloom(HAB)-forming species,produce carbonyl compounds.In this study,we analyzed the carbonyl compounds from eight common HAB-forming species(Akashiwo sanguinea,Karenia mikimotoi,Karlodinium veneficum,Margalefidinium polykrikoides,Prorocentrum donghaiense,P.minimum,Scrippsiella trochoidea,and Heterosig,ma akashiwo)using gas chromatography-mass spectrometry in full scan and selected ion monitoring(SIM)modes.Our results show a ubiquitous presence of carbonyl colmpounds in both dissolved and particulate forms in all species we examined.In the full scan mode,133 chromatographic peaks were detected from 51 samples altogether.Both the varieties of carbonyl compounds and their quantities were algal species dependent,although most of the carbonyl compounds could not be fully identified according to the mass spectral database only due to the unavailability of enough standards for all analytes.Aided with nine standards in SIM mode,we further identified and quantified all nine aldehydes(2-methyl-2-pentenal,trans-2-nonenal,cis-6-nonenal,2,6-dimethyl-5-heptenal,trans-2-hexenal,trans-2-decenal,2,4-heptadienal,trans-trans-2,4-octadienal,and trans-trans-2,4-decadienal).Four of these nine aldehydes were detected in particulate form,which confirmed that trans-2-nonenal could be produced by K.mikimotoi,Karl.veneficum,P.donghaiense,P.minimum,S.trochoidea,and H.akashiwo,2,4-heptadienal and trans-trans-2,4-decadienal by A.sanguinea,M.polykrikoides,and S.trochoidea,and trans,trans-2,4-octadienal by S.trochoidea,respectively.We proved that some dinoflagellate and raphidophyte species could contribute to the pool of carbonyl compounds including PUAs in a marine ecosystem.Some carbonyl compounds,particularly those in high cell quota and/or dissolved concentration,may be related to fishkilling or allelopathy which needs further identification and quantification.     

Effects of algal concentration and initial density on the population growth of Diaphanosoma celebensis Stingelin （Crustacea, Cladocera）

The effects of algal concentration and initial density on the population growth of the estuarine cladocera,Diaphanosoma celebensis Stingelin,were evaluated in an indoor experiment.A 2 × 4 layout that included two algal concentrations(Chlorella pyrenoidosa,1 × 106 and 3 × 106 cell/mL) and four inoculation densities(100,200,300 and 400 ind./L) were established.Diaphanosoma celebensis were reared in 150 mL flasks containing 50 mL of algal medium at 22°C,under salinity of 10 and a photoperiod of 12 h L:12 h D.T...     

南海海域三疣梭子蟹的幼体培育

幼体期Z1和Z2投喂扁藻和轮虫,Z2后期开始以轮虫和卤虫无节幼体混合投喂,Z3后用卤虫无节幼体投喂,大眼幼体用不同饵料培育,进行南海海域三疣梭子蟹幼体培育,结果表明:溞状幼体平均成活率为15.3%;Z2变态为Z3、Z4变态为M为溞状幼体培育的敏感期,幼体的阶段成活率较低;以卤虫无节幼体、冰冻卤虫成虫、活卤虫成虫为饵料,均能使培育大眼幼体至幼蟹,其中活卤虫成虫投喂、加防残网片培育效果最好,成活率达80%。     

A Successful Microbound Diet for the Larval Culture of Chinese Shrimp Fenneropenaeus chinensis

A 13-day feeding trial was carried out to evaluate the effectiveness of a microbound diet for rearing the larvae of Chinese shrimp Fenneropenaeus chinensis in comparison with the live foods that consist of Isochrysis galbana, Chlorella vulgaris, Tetraselrnis chuii, rotifer （Brachionus plicatilis） and Arternia sp. Larvae of 0 to 13d post-hatch （dph） were reared in a temperature-controlled semi-open culture system and stocked at a density of 100 larvae L^-1 in tanks, each containing 50 L sterilized seawater with salinity 30-32. Larvae were manually fed either the live foods or the microbound diet 6 times per day. At 13 dph, the growth of the larvae fed on the microbound diet was approximately 84% of that fed on the live foods. The survival rate of the larvae fed on the microbound diet was 44.29% at 13dph, which was not significantly different from that of larvae fed on live foods （63.55%）. The body length and development index （DI） of the larvae fed on the microbound diet were always lower than those of larvae fed on live foods. However, the differences reached significant levels only at 11 and 13 dph （P〈 0.05）. The mean dry weight loss of the microbound diet was 9.2% after 90 min immersion in seawater, indicating that this diet has a good water stability. The microbound diet contains 52.23% crude protein and 10.27% lipid and is easy to prepare. These characteristics of the diet suggest good potentials for its successful use in the larviculture of other penaeid and fish species.     

环境因子对L型、S型褶皱臂尾轮虫生长的影响

在实验室条件下,通过种群累积培养的方法,研究了不同盐度和小球藻(Chorellavulgaris)、亚心形扁藻(Platymonassubcordiformis)、微绿球藻(Nannochlorisoculata)、波吉卵囊藻(Oocystisborgei)等不同饵料微藻对L型褶皱臂尾轮虫(BrachionusPlicatilistipicus)和S型褶皱臂尾轮虫(BrachionusPlicatilisrotundiformis)生长的影响,探讨这两种轮虫在生态条件上的差异。结果表明:L型和S型褶皱臂尾轮虫对盐度的适应有显著的差异,生长的最适盐度分别为15和20,大水体培养时,其适宜生长的盐度范围分别是15~20和10 ~30。饵料微藻对L型和S型褶皱臂尾轮虫生长的影响一致。小球藻培养效果在4种饵料微藻中最好,其生长率明显高于微绿球藻和亚心形扁藻,波吉卵囊藻培养效果较差。     

Comparative study of hydrographic conditions for algal bloom formation in the coastal waters of east and west of Hong Kong during 1998

Phytoplankton abundance was found to be positively correlated with seasonal changes of seawater temperature in Port Shelter and Lamma Channel, Hong Kong in 1998. Rising water temperature from around 20°C to 25°C coincided with an increase in phytoplankton abundance at both locations. Heavy rains from June to September reduced salinity from 30 to 20, but the decrease in salinity was not correlated with a decline in phytoplankton abundance. In spring 1998, over 0.6×106 cells dm-3 and 0.1×106 cells dm-3 of the...     

Influences of pika and simulated grazing disturbances on bare patches of alpine meadow in the Yellow River Source Zone

Bare patches in alpine meadow are the main manifestation of its degradation. The change of bare patches in an alpine meadow in the Yellow River Source Zone during 2018-2019 was studied in relation to the disturbances caused by plateau pika(Ochotona curzoniae) population and simulated grazing via artificial mowing both independently and interactively. The disturbance was set at three levels of high, medium and no disturbance(control group). Bare patches were mapped by from unmanned aerial vehicle(UAV) images with fine resolution of 1 cm obtained in August 2018 and August 2019 in ArcGIS. The results showed that the total area of bare patches decreased by 112.05 m~2 in sub-plots devoid of pika disturbance but increased by 126.37 m~2 in other subplots. The highest rate of increase is 89.02%. The individual effect of pika exceeds the joint effect of pika and mowing disturbances. The sole effect of mowing is lower than the joint effect of pika disturbance and intensive mowing, but higher than the joint influence of pika disturbance and moderate mowing. Strong pika disturbance(14 per sub-plot) caused the influence of mowing from moderate to intensive to increase by five-fold. The area of bare patches treated with moderate mowing and no pika disturbance decreased at the highest pace(-37.22%). Intensive mowing and medium density pikas(100 pikas/ha) are considered the thresholds at which the bare patches start to expand. Even if the meadow is mowed at the medium and high intensity, the area of bare patches can be significantly reduced if plateau pika population is controlled to a low level. ANOVA analysis and longterm macro-scale satellite-derived results reveal that pika disturbance is more important in causing the bare patches to change than simulated grazing. Therefore, it is more important to control the number of pikas than to reduce grazing intensity to prevent the expansion of bare patches in the degraded alpine meadow in the study area.     

不同条件下波吉卵囊藻对氨氮的吸收速率

在实验室条件下模拟多种氮源共存的虾池水环境,利用稳定同位素示踪剂,研究温度、光照、盐度、pH、藻浓度对波吉卵囊藻氨氮吸收速率的影响。结果表明:温度、光照和藻浓度对波吉卵囊藻氨氮吸收速率影响极显著(P<0.01)。当温度为20℃,照度为4 500 lx,藻浓度为5.5×105/mL,波吉卵囊藻对氨氮均有最大的吸收速率,分别为1.114、1.400、1.482μg g-1 h-1。单因素方差分析得出波吉卵囊藻吸收氨氮的最优组合为:温度20℃,照度4 500lx,盐度15,pH 7.5,藻浓度5.5×105/mL。可通过对环境因子的调控,提高波吉卵囊藻对水体中氨氮的吸收速率。     

Salt response of the freshwater microalga Scenedesmus obliquus (Turp.) Kutz is modulated by the algal growth phase

Growth and biochemical responses of the coenobial green alga Scenedesmus obliquus to salinity stress were monitored across different phases of growth. The alga was cultured on BG11 growth medium and subjected to 0, 30, 100, and 200 mmol/L NaCl for a period of 20 d, during which algal cultures were harvested at 4-d intervals. The salinity-induced inhibition of algal growth was accompanied with prolongation of timing of the different growth phases. The sharp and progressive salinity-induced inhibition of algal growth rate during the early phase of growth points to salt shock but moderation of inhibition at the subsequent stages of growth means algal acclimation to salinity. The concentrations of chlorophylls a and b, soluble sugars, proteins as well as those of K+ and Na+ in the alga exhibited peaks at the initiation of the exponential phase of growth, with increasing magnitude in proportion to the increase in the level of salinity. Nevertheless, whereas soluble sugars of the alga peaked at initiation of the exponential phase, starch concentration progressively increased with culture age, reaching saturation towards the stationary phase. Whereas the salinity-induced increase in soluble sugars was most evident at the early stages of growth the reverse was true for starch. The present results point to fast acclimation of S. obliquus to salt stress post a brief salt shock, utilizing soluble sugars, K+ and Na+ for osmotic adjustment. Increasing salinity from 0 to 200 mmol/L NaCl led to progressive increase in soluble sugars, proteins, K + and Na + concentrations of the algal cells, particularly at the early stages of growth. However, the salinity-induced increase in chlorophyll concentration approached a limit at 100 mmol/L NaCl whereas that in starch concentration was more evident at the later stages of growth.     

Effect of Alexandrium tamarense on three bloom-forming algae

We investigated the allelopathic properties of Alexandrium tamarense (Laboar) Balech on the growth of Prorocentrum donghaiense Lu, Chattonella marina (Subrahmanyan) Hara et Chihara and Heterosigma akashiwo (Hada) Hada in a laboratory experiment. We examined the growth of A. tamarense, C. marina, P. donghaiense and H. Akashiwo in co-cultures and the effect of filtrates from A. tamarense cultures in various growth phases, on the three harmful algal bloom (HAB)-forming algae. In co-cultures with A. tamarense, both C. marina and H. akashiwo were dramatically suppressed at high cell densities; in contrast, the growth of P. donghaiense varied in different inoculative ratios of A. tamarense and P. donghaiense. When the ratio was 1:1 (P. donghaiense: A. tamarense), growth of P. donghaiense was inhibited considerably, while the growth of P. donghaiense was almost the same as that of the control when the ratio was 9:1. The growth difference of P. donghaiense, C. marina and H. akashiwo when co-cultured with A. tamarense indicated that the allelopathic effect may be one of the important factors in algal competition and phytoplankton succession involving A. tamarense. In addition, the filtrate from A. tamarense culture had negative impacts on these three HAB algae, and such inhibition varied with different growth phases of A. tamarense in parallel with reported values of PSP toxin content in Alexandrium cells. This implied that PSP toxin was possibly involved in allelopathy of A. tamarense. However, the rapid decomposition and inactivation of PSP toxin above pH 7 weakened this possibility. Further studies on the allelochemicals responsible for the allelopathy of A. tamarense need to be carried out in future.     

A successful microbound diet for the larval culture of Chinese shrimp Fenneropenaeus chinensis

A 13-day feeding trial was carried out to evaluate the effectiveness of a microbound diet for rearing the larvae of Chinese shrimp Fenneropenaeus chinensis in comparison with the live foods that consist of Isochrysis galbana, Chlorella vulgaris, Tetraselmis chuii, rotifer (Brachionus plicatilis) and Artemia sp. Larvae of 0 to 13d post-hatch (dph) were reared in a temperature-controlled semi-open culture system and stocked at a density of 100 larvae L⁻¹ in tanks, each containing 50 L sterilized seawater with salinity 30–32. Larvae were manually fed either the live foods or the microbound diet 6 times per day. At 13dph, the growth of the larvae fed on the microbound diet was approximately 84% of that fed on the live foods. The survival rate of the larvae fed on the microbound diet was 44.29% at 13 dph, which was not significantly different from that of larvae fed on live foods (63.55% ). The body length and development index (DI) of the larvae fed on the microbound diet were always lower than those of larvae fed on live foods. However, the differences reached significant levels only at 11 and 13dph (P<0.05). The mean dry weight loss of the microbound diet was 9.2% after 90min immersion in seawater, indicating that this diet has a good water stability. The microbound diet contains 5223% crude protein and 10.27% lipid and is easy to prepare. These characteristics of the diet suggest good potentials for its successful use in the larviculture of other penaeid and fish species.