Karenia mikimotoi (Miyake & Kominami ex Oda) Hansen & Moestrup is associated with harmful algal blooms in temperate and subtropical zones of the world. The hemolytic substances produced by K. mikimotoi are thought...Karenia mikimotoi (Miyake & Kominami ex Oda) Hansen & Moestrup is associated with harmful algal blooms in temperate and subtropical zones of the world. The hemolytic substances produced by K. mikimotoi are thought to cause mortality in fishes and invertebrates. We evaluated the composition of the hemolytic toxin produced by K. mikimotoi cultured in the laboratory using thin-layer chromatography. In addition, we evaluated the effect of co-occuring algae (Prorocentrum donghaiense and Alexandrium tamarense) and the cladoceran grazer Moina mongoliea on hemolytic toxin production in K. mikimotoi. The hemolytic toxins from K. mikimotoi were a mixture of 2 liposaccharides and 1 lipid. Waterborne clues from P. donghaiense and A. tamarense inhibited the growth of K. mikimotoi but increased the production of hemolytic toxins. Conversely, K. mikimotoi strongly inhibited the growth of caged P. donghaiense and A. tamarense. In addition, the ingestion of K. mikimotoi by M. mongolica induced the production of hemolytic toxins in K. mikimotoi. Taken together, our results suggest that the presence of other microalgae and grazers may be as important as environmental factors for controlling the production of hemolytic substances. K. mikimotoi secreted allelochemicals other than unstable fatty acids with hemolytic activity. The production of hemolytic toxins in dinofiagellates was not only dependent on resource availability, but also on the risk of predation. Hemolytic toxins likely play an important role as chemical deterrents secreted by K. mikimotoi.展开更多
Harmful algal blooms(HABs) have led to extensive ecological and environmental issues and huge economic losses.Various HAB control techniques have been developed,and biological methods have been paid more attention.Alg...Harmful algal blooms(HABs) have led to extensive ecological and environmental issues and huge economic losses.Various HAB control techniques have been developed,and biological methods have been paid more attention.Algicidal bacteria is a general designation for bacteria which inhibit algal growth in a direct or indirect manner,and kill or damage the algal cells.A metabolite which is strongly toxic to the dinoflagellate Alexandrium tamarense was produced by strain DH46 of the alga-lysing bacterium Alteromonas sp.The culture conditions were optimized using a single-factor test method.Factors including carbon source,nitrogen source,temperature,initial pH value,rotational speed and salinity were studied.The results showed that the cultivation of the bacteria at 28℃ and 180 r min-1with initial pH 7 and 30 salt contcentration favored both the cell growth and the lysing effect of strain DH46.The optimal medium composition for strain DH46 was determined by means of uniform design experimentation,and the most important components influencing the cell density were tryptone,yeast extract,soluble starch,NaNO3 and MgSO4.When the following culture medium was used(tryptone 14.0g,yeast extract 1.63g,soluble starch 5.0 g,NaNO3 1.6 g,MgSO4 2.3 g in 1L),the largest bacterial dry weight(7.36 g L-1) was obtained,which was an enhancement of 107% compared to the initial medium;and the algal lysis rate was as high as 98.4% which increased nearly 10% after optimization.展开更多
Rapid,high-throughput and reliable methods are urgently required to accurately detect and monitor harmful algae,which are responsible for algal blooms,such as red and green tides. In this study,we successfully develop...Rapid,high-throughput and reliable methods are urgently required to accurately detect and monitor harmful algae,which are responsible for algal blooms,such as red and green tides. In this study,we successfully developed a multiplex PCR-based DNA microarray method capable of detecting nine harmful algal species simultaneously,namely A lexandrium tamarense,Gyrodinium instriatum,Heterosigma akashiwo,Karenia mikimotoi,Prorocentrum donghaiense,Prorocentrum minimum,Ulva compressa,Ulva ohnoi and Ulva prolifera. This method achieved a limit of detection(LOD) of 0.5 ng of genomic DNA(orders of magnitude of the deci-nanogram range) in the tested algae cultures. Altogether,230 field samples from ship ballast waters and seaport waters were used to evaluate the DNA microarray. The clinical sensitivity and specificity of the DNA microarray assay in detecting field samples were 96.4% and 90.9%,respectively,relative to conventional morphological methods. This indicated that this high-throughput,automatic,and specific method is well suited for the detection of algae in water samples.展开更多
The impact of harmful algal blooms (HABs) on public health and related economics have been increasing in many coastal regions of the world. Sedimentation of algal cells through flocculation with clay particles is a ...The impact of harmful algal blooms (HABs) on public health and related economics have been increasing in many coastal regions of the world. Sedimentation of algal cells through flocculation with clay particles is a promising strategy for controlling HABs. Previous studies found that removal efficiency (RE) was influenced by many factors, including clay type and concentration, algal growth stage, and physiological aspects of HAB cells. To estimate the effect of morphological characteristics of the aggregates on HAB cell removal, fractal dimensions were measured and the RE of three species of HAB organism, Heterosigma akashiwo, Alexandrium tamarense, and Skeletonema eostatum, by original clay and modified clay, was determined. For all HAB species, the modified clay had a higher RE than original clay. For the original clay, the two-dimensional fractal dimension (D2) was 1.92 and three-dimensional ffactal dimension (D3) 2.81, while for the modified clay, D2 was 1.84 and D3 was 2.50. The addition of polyaluminum chloride (PAC1) lead to a decrease of the repulsive barrier between clay particles, and resulted in lower D2 and D3. Due to the decrease of D3, and the increase of the effective sticking coefficient, the flocculation rate between modified clay particles and HAB organisms increased, and thus resulted in a high RE. The fractal dimensions of flocs differed in HAB species with different cell morphologies. For example, Alexandrium tamarense cells are ellipsoidal, and the D3 and D2 of flocs were the highest, while for Skeletonema costatum, which has filamentous cells, the D3 and D2 of flocs were the lowest.展开更多
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 ...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 pH7 weakened this possibility.Further studies on the allelochemicals responsible for the allelopathy of A.tamarense need to be carried out in future.展开更多
This study investigated the inductive effect ofAlexandrium tamarense, a toxic dinoflagellate producing paralytic shell- fish poison, on oxidative stress and apoptosis in hepatopancreas of Chinese shrimp, Fenneropenaeu...This study investigated the inductive effect ofAlexandrium tamarense, a toxic dinoflagellate producing paralytic shell- fish poison, on oxidative stress and apoptosis in hepatopancreas of Chinese shrimp, Fenneropenaeus chinensis. The individuals of E chinensis were exposed to 200 and 1000 cells mL-1 of A. tamarense with their superoxide dismutase (SOD), glutathione S-transferase (GST) activities, malonyldialdehyde (MDA) concentration, and caspase gene (FcCasp) expression in hepatopancreas determined at 12, 24, 48, 72 and 96 h. In addition, apoptosis in hepatopancreas of E chinensis at 96 h after exposure was determined through terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay. The hepatopancreatic SOD and GST activities of F. chinensis exposed to 1000 cells mL-1 ofA. tamarense showed a bell-shaped response to exposure time. The hepatopancreatic MDA concentration ofF. chinensis exposed to 1000 cellsmL-1 ofA. tamarense increased gradually from 48 to 96h, and such a trend corresponded to the decrease of GST activity. The hepatopancreatic FcCasp transcript abundance of F. chinensis exposed to 1000 cells mL-1 ofA. tamarense was positively and linearly correlated to MDA concentration. Results of TUNEL assay showed that exposure to 1000 cells mL-1 of A. tamarense induced apoptosis in the hepatopancreas of E chinensis. Our study revealed that A. tamarense exposure influenced the antioxidative status ofF. chinensis and caused lipid peroxidation and apoptosis in the hepatopancreas of shrimp.展开更多
Overthelastseveraldecades,harmfulalgalblooms(HABs)havebecomeaseriousenvironmental problem in many parts of the world. A rapid and accurate detection process for HAB algae has yet to be developed. Heterosigma akashiwo ...Overthelastseveraldecades,harmfulalgalblooms(HABs)havebecomeaseriousenvironmental problem in many parts of the world. A rapid and accurate detection process for HAB algae has yet to be developed. Heterosigma akashiwo is one of the most important HABs species in China. The objective of this study was to develop an immunologic technique that can rapidly and sensitively count H. akashiwo cells. Five HABs species (Alexandrium catenella, Thalassiosira sp., Cryptomonas sp., Alexandrium tamarense and Symbiodinium sp.,) were used in this study to evaluate the analysis process we developed. A polyclonal antibody with high titers against H. akashiwo was obtained by injecting H. akashiwo cells into rabbits. Immuno-magnetic beads (IMB) were produced via conjugated polyclonal antibodies with magnetic beads and applied to isolate and count H. akashiwo cells from the culture. Results show that 66.7%-91.6% of the cells were captured from unialgal culture by IMBs, and only 5.3%-12.5% of the four other HAB microalgae species were captured, indicating that the constructed IMBs combined specifically with the H. akashiwo cells. At the same time, flow cytometry (FCM) sorting was exploited to screen H. akashiwo cells after labeling with FITC conjugated polyclonal antibodies. Using the FCM technique, 91.7% of the targeted cells were sorted out from mixed microalgae samples in just a few minutes. These results indicate that both antibody-involved IMB and antibody-based FCM techniques are highly effective at detecting and quantifying HAB species. These techniques, especially immuno-magnetic separation, have low associated cost, and are fast and simple processes compared with other techniques currently in use.展开更多
基金Supported by the National Natural Science Foundation of China(Nos.30970502,U0733006)the National Basic Research Program of China(973 Program)(No.2010CB428702)
文摘Karenia mikimotoi (Miyake & Kominami ex Oda) Hansen & Moestrup is associated with harmful algal blooms in temperate and subtropical zones of the world. The hemolytic substances produced by K. mikimotoi are thought to cause mortality in fishes and invertebrates. We evaluated the composition of the hemolytic toxin produced by K. mikimotoi cultured in the laboratory using thin-layer chromatography. In addition, we evaluated the effect of co-occuring algae (Prorocentrum donghaiense and Alexandrium tamarense) and the cladoceran grazer Moina mongoliea on hemolytic toxin production in K. mikimotoi. The hemolytic toxins from K. mikimotoi were a mixture of 2 liposaccharides and 1 lipid. Waterborne clues from P. donghaiense and A. tamarense inhibited the growth of K. mikimotoi but increased the production of hemolytic toxins. Conversely, K. mikimotoi strongly inhibited the growth of caged P. donghaiense and A. tamarense. In addition, the ingestion of K. mikimotoi by M. mongolica induced the production of hemolytic toxins in K. mikimotoi. Taken together, our results suggest that the presence of other microalgae and grazers may be as important as environmental factors for controlling the production of hemolytic substances. K. mikimotoi secreted allelochemicals other than unstable fatty acids with hemolytic activity. The production of hemolytic toxins in dinofiagellates was not only dependent on resource availability, but also on the risk of predation. Hemolytic toxins likely play an important role as chemical deterrents secreted by K. mikimotoi.
基金financially supported by the National Natural Science Foundation(40930847,31070442)the Natural Science Foundation of Fujian Province(2012J01150)Public science and technology research funds projects of ocean(201305016,201305041,201305022) and MELRI1003
文摘Harmful algal blooms(HABs) have led to extensive ecological and environmental issues and huge economic losses.Various HAB control techniques have been developed,and biological methods have been paid more attention.Algicidal bacteria is a general designation for bacteria which inhibit algal growth in a direct or indirect manner,and kill or damage the algal cells.A metabolite which is strongly toxic to the dinoflagellate Alexandrium tamarense was produced by strain DH46 of the alga-lysing bacterium Alteromonas sp.The culture conditions were optimized using a single-factor test method.Factors including carbon source,nitrogen source,temperature,initial pH value,rotational speed and salinity were studied.The results showed that the cultivation of the bacteria at 28℃ and 180 r min-1with initial pH 7 and 30 salt contcentration favored both the cell growth and the lysing effect of strain DH46.The optimal medium composition for strain DH46 was determined by means of uniform design experimentation,and the most important components influencing the cell density were tryptone,yeast extract,soluble starch,NaNO3 and MgSO4.When the following culture medium was used(tryptone 14.0g,yeast extract 1.63g,soluble starch 5.0 g,NaNO3 1.6 g,MgSO4 2.3 g in 1L),the largest bacterial dry weight(7.36 g L-1) was obtained,which was an enhancement of 107% compared to the initial medium;and the algal lysis rate was as high as 98.4% which increased nearly 10% after optimization.
基金Supported by the National High Technology Research and Development Program of China(863 Program)(No.2012AA092001)the Science and Technology Project of Zhejiang Province(No.2013C03045-1)+2 种基金the Zhejiang Marine Biotechnology Innovation Team(No.2010R50029-12)the Natural Science Foundation of Ningbo City of China(No.2013A610168)the KC Wong Magna Fund in Ningbo University
文摘Rapid,high-throughput and reliable methods are urgently required to accurately detect and monitor harmful algae,which are responsible for algal blooms,such as red and green tides. In this study,we successfully developed a multiplex PCR-based DNA microarray method capable of detecting nine harmful algal species simultaneously,namely A lexandrium tamarense,Gyrodinium instriatum,Heterosigma akashiwo,Karenia mikimotoi,Prorocentrum donghaiense,Prorocentrum minimum,Ulva compressa,Ulva ohnoi and Ulva prolifera. This method achieved a limit of detection(LOD) of 0.5 ng of genomic DNA(orders of magnitude of the deci-nanogram range) in the tested algae cultures. Altogether,230 field samples from ship ballast waters and seaport waters were used to evaluate the DNA microarray. The clinical sensitivity and specificity of the DNA microarray assay in detecting field samples were 96.4% and 90.9%,respectively,relative to conventional morphological methods. This indicated that this high-throughput,automatic,and specific method is well suited for the detection of algae in water samples.
基金Supported by the Fund for Creative Research Groups by National Natural Science Foundation of China (No. 40821004)the National Natural Science Foundation of China (No. 40906055)the National Basic Research Program of China (973 Program) (No. 2010CB428706)
文摘The impact of harmful algal blooms (HABs) on public health and related economics have been increasing in many coastal regions of the world. Sedimentation of algal cells through flocculation with clay particles is a promising strategy for controlling HABs. Previous studies found that removal efficiency (RE) was influenced by many factors, including clay type and concentration, algal growth stage, and physiological aspects of HAB cells. To estimate the effect of morphological characteristics of the aggregates on HAB cell removal, fractal dimensions were measured and the RE of three species of HAB organism, Heterosigma akashiwo, Alexandrium tamarense, and Skeletonema eostatum, by original clay and modified clay, was determined. For all HAB species, the modified clay had a higher RE than original clay. For the original clay, the two-dimensional fractal dimension (D2) was 1.92 and three-dimensional ffactal dimension (D3) 2.81, while for the modified clay, D2 was 1.84 and D3 was 2.50. The addition of polyaluminum chloride (PAC1) lead to a decrease of the repulsive barrier between clay particles, and resulted in lower D2 and D3. Due to the decrease of D3, and the increase of the effective sticking coefficient, the flocculation rate between modified clay particles and HAB organisms increased, and thus resulted in a high RE. The fractal dimensions of flocs differed in HAB species with different cell morphologies. For example, Alexandrium tamarense cells are ellipsoidal, and the D3 and D2 of flocs were the highest, while for Skeletonema costatum, which has filamentous cells, the D3 and D2 of flocs were the lowest.
基金Supported by the NSFC-Guangdong Province Association Foundation(No.U0733006)the National Natural Science Foundation of China(No.30970502,40976065)the National Basic Research Program of China(973 Program)(No.2010CB428702)
文摘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 pH7 weakened this possibility.Further studies on the allelochemicals responsible for the allelopathy of A.tamarense need to be carried out in future.
基金supported by Earmarked Fund for Modern Agro-industry Technology Research System of China (Grant No. CARS-47)Special Fund for Agroscientific Research in the Public Interest of China (Grant No. 201103034)the National ‘863’ Project of China (Grant No. 2012AA10A409)
文摘This study investigated the inductive effect ofAlexandrium tamarense, a toxic dinoflagellate producing paralytic shell- fish poison, on oxidative stress and apoptosis in hepatopancreas of Chinese shrimp, Fenneropenaeus chinensis. The individuals of E chinensis were exposed to 200 and 1000 cells mL-1 of A. tamarense with their superoxide dismutase (SOD), glutathione S-transferase (GST) activities, malonyldialdehyde (MDA) concentration, and caspase gene (FcCasp) expression in hepatopancreas determined at 12, 24, 48, 72 and 96 h. In addition, apoptosis in hepatopancreas of E chinensis at 96 h after exposure was determined through terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay. The hepatopancreatic SOD and GST activities of F. chinensis exposed to 1000 cells mL-1 ofA. tamarense showed a bell-shaped response to exposure time. The hepatopancreatic MDA concentration ofF. chinensis exposed to 1000 cellsmL-1 ofA. tamarense increased gradually from 48 to 96h, and such a trend corresponded to the decrease of GST activity. The hepatopancreatic FcCasp transcript abundance of F. chinensis exposed to 1000 cells mL-1 ofA. tamarense was positively and linearly correlated to MDA concentration. Results of TUNEL assay showed that exposure to 1000 cells mL-1 of A. tamarense induced apoptosis in the hepatopancreas of E chinensis. Our study revealed that A. tamarense exposure influenced the antioxidative status ofF. chinensis and caused lipid peroxidation and apoptosis in the hepatopancreas of shrimp.
基金Supported by Specialized Research Fund for the Doctoral Program of Higher Education of China (No. SRFDP 20100132110007)Shandong Provincial Natural Science Foundation, China (No. ZR2011DZ002)
文摘Overthelastseveraldecades,harmfulalgalblooms(HABs)havebecomeaseriousenvironmental problem in many parts of the world. A rapid and accurate detection process for HAB algae has yet to be developed. Heterosigma akashiwo is one of the most important HABs species in China. The objective of this study was to develop an immunologic technique that can rapidly and sensitively count H. akashiwo cells. Five HABs species (Alexandrium catenella, Thalassiosira sp., Cryptomonas sp., Alexandrium tamarense and Symbiodinium sp.,) were used in this study to evaluate the analysis process we developed. A polyclonal antibody with high titers against H. akashiwo was obtained by injecting H. akashiwo cells into rabbits. Immuno-magnetic beads (IMB) were produced via conjugated polyclonal antibodies with magnetic beads and applied to isolate and count H. akashiwo cells from the culture. Results show that 66.7%-91.6% of the cells were captured from unialgal culture by IMBs, and only 5.3%-12.5% of the four other HAB microalgae species were captured, indicating that the constructed IMBs combined specifically with the H. akashiwo cells. At the same time, flow cytometry (FCM) sorting was exploited to screen H. akashiwo cells after labeling with FITC conjugated polyclonal antibodies. Using the FCM technique, 91.7% of the targeted cells were sorted out from mixed microalgae samples in just a few minutes. These results indicate that both antibody-involved IMB and antibody-based FCM techniques are highly effective at detecting and quantifying HAB species. These techniques, especially immuno-magnetic separation, have low associated cost, and are fast and simple processes compared with other techniques currently in use.