Suitable temperature and light intensity play important roles in the formation of harmful algae blooms(HABs),which can pose serious threats to aquatic ecosystems and human health.In this study,we measured the growth,p...Suitable temperature and light intensity play important roles in the formation of harmful algae blooms(HABs),which can pose serious threats to aquatic ecosystems and human health.In this study,we measured the growth,physiological function,and paralytic shellfish toxins(PSTs)production of Alexandrium pacificum(CCMA-272),a strain isolated from East China Sea,at different temperatures(15,20,and 25℃)and light intensities(30,60,and 90μmol photons/(m^(2)·s)).Results indicate that temperature and light intensity significantly affected the growth,physiology,and toxigenic potentials of A.pacificum.The optimal conditions for the growth of A.pacificum were observed at 20℃ under60μmol photons/(m^(2)·s).Regarding the production of PSTs,this strain of A.pacificum produced 12 PSTs,including carbamate toxins:saxitoxin(STX),neosaxitoxin(NEO),and gonyautoxin 1–4(GTX1,GTX2,GTX3,GTX4);dicarbamoyl toxins:dicarbamoylsaxitoxin(dcSTX),dicarbamoylgonyautoxin 2,3(dcGTX2,dcGTX3);and N-sulfocarbamoyl toxins:N-sulfocarbamoylgonyautoxin 1,2(C1,C2),and gonyautoxin 5(GTX5).Among all the PSTs,C2 was the most abundant.Low temperature(15℃)and high light intensity(90μmol photons/(m^(2)·s))were beneficial for the production of PSTs in A.pacificum.When cultured at 20 and 25℃,A.pacificum generated comparable total quantities of PSTs,yet the toxicity levels were lower at 25℃.Intra-cellular PSTs contents were greater than extra-cellular PSTs contents,except those under the condition of 25℃ with 30μmol photons/(m^(2)·s).However,as the increase of temperature,A.pacificum released more amounts of analogues with higher toxicity levels(e.g.,STX and dcGTX_(2))into the environment than intracellularly.These findings emphasize the significant sensitivity of A.pacificum to temperature and light intensity,highlighting the importance of evaluating both intra-cellular and extra-cellular PSTs for assessing its toxicity and aiding in the prediction and management of HABs.展开更多
Alexandrium pacificum(A.pacificum)is a typical paralytic shellfish poisonous dinoflagellate.Harmful algal blooms(HABs)caused by this species can bring serious environmental problems and economic losses to the aquacult...Alexandrium pacificum(A.pacificum)is a typical paralytic shellfish poisonous dinoflagellate.Harmful algal blooms(HABs)caused by this species can bring serious environmental problems and economic losses to the aquaculture industry.In this study,transcriptome sequencing and analyses were performed on the neural tissue of Litopenaeus vannamei(L.vannamei)after acute exposure to A.pacificum disrupted solution for 72 h,and differentially expressed genes(DEGs)were identified.The results showed that,compared with the control samples,300 DEGs were identified in the experimental group,of which 194 were up-regulated,and 106 down-regulated.The gene ontology(GO)functional enrichment analysis showed that DEGs were significantly enriched in the cortical cytoskeleton organization,troponin complex,amylo-alpha-1,6-glucosidase and thymidine phosphorylase.Kyoto encyclopedia of genes and genomes(KEGG)enrichment analysis found that DEGs were mainly enriched in the oxidative phosphorylation process,intercellular tight junctions and mitophagy.The results showed that the proteoglycans,signaling pathways,and various metabolic processes that regulate cell proliferation,differentiation,and apoptosis all played an essential role in the response of L.vannamei to A.pacificum toxins.展开更多
Aim To study isolation and purification of Gonyautoxins from two differentstrains of Alexandrium minutum Halim. Methods Ethanol extracts of the two culture strains ofAlexandrium minutum Halim were fractionated by gel ...Aim To study isolation and purification of Gonyautoxins from two differentstrains of Alexandrium minutum Halim. Methods Ethanol extracts of the two culture strains ofAlexandrium minutum Halim were fractionated by gel filtration and ion exchange chromatography toobtain gonyautoxins. The toxin profiles were tested by HPLC method. Results Both strains containgonyautoxin-1, gonyautoxin-2, gonyautoxin-3 and gonyautoxin-4. The total gonyautoxin content ofAmtk4 strain is almost 10 times higher than that of Amtk2 strain. Conclusion Alexandrium minutumHalim Amtk4 strain is suitable for the preparation of Gonyautoxins.展开更多
A loop-mediated isothermal amplification （LAMP） assay was designed and evaluated for rapid de- tection of the toxic microalgae Alexandrium catenella and A. minutum, which can produce paralytic shellfish poisoning （...A loop-mediated isothermal amplification （LAMP） assay was designed and evaluated for rapid de- tection of the toxic microalgae Alexandrium catenella and A. minutum, which can produce paralytic shellfish poisoning （PSP）. Two sets of four specific primers targeting these two species were derived from the sequence of internal transcribed spacer （ITS） of ribosomal DNA. The method worked well in less than an hour under isothermal conditions of 65℃. LAMP specificity was validated in closely related algae as a comparison, suggesting the strict specificity of the LAMP primers. Two visual inspection approaches were feasible to interpret the positive or negative results. The detection lim- its of A. catenella and A. minutum samples using the LAMP assay were found to be 5.6 and 4.5 pg DNA, respectively. The sensitivity of this LAMP assay was 10 or 100-fold higher than Polymerase Chain Reaction （PCR） method in detecting the two microalgae. These characteristics of species specificity, sensitivity, and rapidity suggest that this method has the potentiality in the monitoring of red tide caused by A. catenella and A. minutum.展开更多
Enhanced UV-B(280-320 nm) radiation resulting from ozone depletion is one of global environmental problems. Not only marine organisms but also marine ecosystems can be affected by enhanced UV-B radiation. The effects ...Enhanced UV-B(280-320 nm) radiation resulting from ozone depletion is one of global environmental problems. Not only marine organisms but also marine ecosystems can be affected by enhanced UV-B radiation. The effects of UV-B radiation on interaction of macro-algae and micro-algae were investigated using Ulva pertusa Kjellman and Alexandrium tamarense as the materials in this study. The results demonstrated that UV-B radiation could inhibit the growth of Ulva pertusa and Alexandrium tamarense when they were both mono-cultured, and the growth inhibition of algae was more significant with increasing doses of UV-B radiation. Alexandrium tamarense could inhibit the growth of Ulva pertusa in mixed culture, and the growth inhibition was more significant when increasing the initial cell density. However, Ulva pertusa could inhibit the growth of Alexandrium tamarense in early phase and stimulate the growth in latter phase when they were grown in mixed culture. Lower initial cell density(10~2 cell/ml) of Alexandrium tamarense could inhibit the growth of Ulva pertusa under UV-B radiation treatment,however, with the initial cell density increasing(10~3 and 10~4 cell/ml), the growth of Ulva pertusa was stimulated under lower dose of UV-B radiation and inhibited under higher dose of UV-B radiation by Alexandrium tamarense. Compared with that in mixed culture, Ulva pertusa showed more positive inhibition to the growth of Alexandrium tamarense under UV-B radiation treatment.展开更多
Paralytic shellfish poisoning (PSP) toxins are potent neurotoxins mainly produced by dinoflagellates and being concentrated in bivalves through food web transfer. Increasing number of findings of toxin-producing bacte...Paralytic shellfish poisoning (PSP) toxins are potent neurotoxins mainly produced by dinoflagellates and being concentrated in bivalves through food web transfer. Increasing number of findings of toxin-producing bacteria in the cells of dinoflagellate such as Alexandriumtamarense supports the hypothesis of the bacterial origin of PSP toxins. Evidence that there are specific symbiosis bacterial taxa associated with the phytoplankton indicates the presence of specific selective mechanisms between them, and implies that the symbiosis bacteria have some vital function to the benefit of the dinoflagellates. Studies on the role of toxin-producing symbiosis bacteria in the marine ecosystem are considered to be becoming more important. Although toxigenic bacteria could be isolated from toxic dinoflagellates, it was not clearly proven whether the isolated bacterial strains based on culture-dependent manner and the corresponding intracellular bacteria were the same because of microbial unculturability. This paper aims to demonstrate the biodiversity of the symbiotic bacteria associated with toxic dinoflagellate A. tamarense using the culture-indepen- dent high-throughput pyrosequencing method, as well as the phylogenetic analysis based on 16S rDNA sequences of the symbiotic cultivable bacteria strains isolated from toxic Alexander tamarense.展开更多
Objective Impact of the presence of bacteria associated with a marine dinoflagellate, Alexandrium tamarense CI01, on the growth and toxin production of the algae in batch culture was investigated. Methods Pronounced c...Objective Impact of the presence of bacteria associated with a marine dinoflagellate, Alexandrium tamarense CI01, on the growth and toxin production of the algae in batch culture was investigated. Methods Pronounced changes in the activities of the algal culture were observed when the culture was treated with different doses of a mixture of penicillin and streptomycin. Results In the presence of antibiotics at the initial concentration of 100 u/mL in culture medium, both algal growth and toxin yield increased markedly. When the concentration of antibiotics was increased to 500 u/mL, the microalgal growth was inhibited, but resumed in a few days to eventually reach the same level of growth and toxin production as at the lower dose of the antibiotics. When the antibiotics were present at a concentration of 1 000 u/mL, the algal growth was inhibited permanently. Conclusions The results indicate that antibiotics can enhance algal growth and toxin production not only through their inhibition of the growth and hence competition for nutrients, but also through their effects on the physiology of the algae.展开更多
The effects of a PSP producing dinoflagellate Alexandrium tamarense on marine bivalvesat their several important life stages: egg, D- shape larva, eyespot larva, juvenile and adult, were studied. The results show that...The effects of a PSP producing dinoflagellate Alexandrium tamarense on marine bivalvesat their several important life stages: egg, D- shape larva, eyespot larva, juvenile and adult, were studied. The results show that the hatching, survival, activity, filtration and growth were adversely affected by the alga and the impact was significantly increased with the increase of algal density. The inhibitory effect on egg hatching was most significant, which the hatching rate was only 30% of the control when exposed to the alga at 100 cell/cm3 after 36 h. Further experiments show that the algal culture, re-suspended cells and cell fragments had the inhibitory effect, while no such effect was from the cell-free medium, cell contents and standard STX. The results indicate that the alga could produce unknown toxins, rather than PSP, associated with the cell surface.展开更多
The effect of Ca 2+ on the removal of Alexandrium sp. LC3 under HDTMAB stress was investigated. The results showed that the toxic effect of HDTMAB on Alexandrium sp. LC3 was significantly reduced in the presence of C...The effect of Ca 2+ on the removal of Alexandrium sp. LC3 under HDTMAB stress was investigated. The results showed that the toxic effect of HDTMAB on Alexandrium sp. LC3 was significantly reduced in the presence of Ca 2+, especially under 4 mmol/L of Ca 2+. To understand the underlying mechanism, the SH group and MDA content of the cell membrane and membrane permeability were measured. It was found that the SH content of cell member increased, the MDA content and membrane permeability decreased when Alexandrium sp. was treated with Ca 2+ and HDTMAB complex, compared with using HDTMAB only. The data suggested that Ca 2+ might promote HDTMAB stress resistance of Alexandrium sp. LC3 by reducing the permeability and increasing the stability of cell membrane.展开更多
Harmful algal blooms （HABs） have been affecting negatively the shellfish and aquaculture industries around the world. Though a lot of efforts have been made to disclose the changes of environmental factors involved ...Harmful algal blooms （HABs） have been affecting negatively the shellfish and aquaculture industries around the world. Though a lot of efforts have been made to disclose the changes of environmental factors involved and their effects on the HABs events, the molecular mechanism of this process remains unclear. To address this problem, proliferating cell nuclear antigen gene （pcna） was isolated and characterized from Alexandrium catenella. It showed high homology to those of other dinoflagellates （89% and 91% homology to Pfiesteria piscicid and Pyrocystis lunula, respectively）, and also 42%–43% homology to those of plant and animals. The expression level of pcna revealed by quantitative real time PCR was the lowest at the late lagging cell growth phase, increased to the highest at the late exponential phase, and then decreased at the stationary phase. Though the cell growth rate was also changing, no positive correlation between pcna expression level and cell growth rate was displayed throughout the whole cell growth stages （r 2 =0.024 6）. However, the pcna expression level had the similar trend with the change of cell growth rate throughout the whole growing process, e.g., from increasing at the earlier cell growth stage to decreasing at the following stages, though slightly lagging to the latter.展开更多
基金Supported by the National Natural Science Foundation of China(Nos.32101290,52009082)the Natural Science Foundation of Jiangsu Province(No.BK20210364)the Fundamental Research Funds for the Central Universities(No.B220202044)。
文摘Suitable temperature and light intensity play important roles in the formation of harmful algae blooms(HABs),which can pose serious threats to aquatic ecosystems and human health.In this study,we measured the growth,physiological function,and paralytic shellfish toxins(PSTs)production of Alexandrium pacificum(CCMA-272),a strain isolated from East China Sea,at different temperatures(15,20,and 25℃)and light intensities(30,60,and 90μmol photons/(m^(2)·s)).Results indicate that temperature and light intensity significantly affected the growth,physiology,and toxigenic potentials of A.pacificum.The optimal conditions for the growth of A.pacificum were observed at 20℃ under60μmol photons/(m^(2)·s).Regarding the production of PSTs,this strain of A.pacificum produced 12 PSTs,including carbamate toxins:saxitoxin(STX),neosaxitoxin(NEO),and gonyautoxin 1–4(GTX1,GTX2,GTX3,GTX4);dicarbamoyl toxins:dicarbamoylsaxitoxin(dcSTX),dicarbamoylgonyautoxin 2,3(dcGTX2,dcGTX3);and N-sulfocarbamoyl toxins:N-sulfocarbamoylgonyautoxin 1,2(C1,C2),and gonyautoxin 5(GTX5).Among all the PSTs,C2 was the most abundant.Low temperature(15℃)and high light intensity(90μmol photons/(m^(2)·s))were beneficial for the production of PSTs in A.pacificum.When cultured at 20 and 25℃,A.pacificum generated comparable total quantities of PSTs,yet the toxicity levels were lower at 25℃.Intra-cellular PSTs contents were greater than extra-cellular PSTs contents,except those under the condition of 25℃ with 30μmol photons/(m^(2)·s).However,as the increase of temperature,A.pacificum released more amounts of analogues with higher toxicity levels(e.g.,STX and dcGTX_(2))into the environment than intracellularly.These findings emphasize the significant sensitivity of A.pacificum to temperature and light intensity,highlighting the importance of evaluating both intra-cellular and extra-cellular PSTs for assessing its toxicity and aiding in the prediction and management of HABs.
基金supported by the Modern Seed Industry Park for Whiteleg Shrimp of Guangdong Province(No.K22226)the National Natural Science Foundation of China(No.32102796)+3 种基金the Natural Science Foundation of Guangdong Province(No.2020A1515110086)the Program for Scientific Research Start-up Funds of Guangdong Ocean University(Nos.060302022102,060302022201)the Program of Shrimp Aquaculture Talent Development(No.B22424)the Undergraduate Innovation Team of Guangdong Ocean University(No.CXTD2023002).
文摘Alexandrium pacificum(A.pacificum)is a typical paralytic shellfish poisonous dinoflagellate.Harmful algal blooms(HABs)caused by this species can bring serious environmental problems and economic losses to the aquaculture industry.In this study,transcriptome sequencing and analyses were performed on the neural tissue of Litopenaeus vannamei(L.vannamei)after acute exposure to A.pacificum disrupted solution for 72 h,and differentially expressed genes(DEGs)were identified.The results showed that,compared with the control samples,300 DEGs were identified in the experimental group,of which 194 were up-regulated,and 106 down-regulated.The gene ontology(GO)functional enrichment analysis showed that DEGs were significantly enriched in the cortical cytoskeleton organization,troponin complex,amylo-alpha-1,6-glucosidase and thymidine phosphorylase.Kyoto encyclopedia of genes and genomes(KEGG)enrichment analysis found that DEGs were mainly enriched in the oxidative phosphorylation process,intercellular tight junctions and mitophagy.The results showed that the proteoglycans,signaling pathways,and various metabolic processes that regulate cell proliferation,differentiation,and apoptosis all played an essential role in the response of L.vannamei to A.pacificum toxins.
文摘Aim To study isolation and purification of Gonyautoxins from two differentstrains of Alexandrium minutum Halim. Methods Ethanol extracts of the two culture strains ofAlexandrium minutum Halim were fractionated by gel filtration and ion exchange chromatography toobtain gonyautoxins. The toxin profiles were tested by HPLC method. Results Both strains containgonyautoxin-1, gonyautoxin-2, gonyautoxin-3 and gonyautoxin-4. The total gonyautoxin content ofAmtk4 strain is almost 10 times higher than that of Amtk2 strain. Conclusion Alexandrium minutumHalim Amtk4 strain is suitable for the preparation of Gonyautoxins.
基金The Science and Technology Commission of Shanghai Municipality under contract Nos 06235810108DZ1980802 and 10JC1418600a special research fund for the national non-profit institutes (East China Sea Fisheries Research Institute) under contract Nos 2007M22 and 2007Z01
文摘A loop-mediated isothermal amplification （LAMP） assay was designed and evaluated for rapid de- tection of the toxic microalgae Alexandrium catenella and A. minutum, which can produce paralytic shellfish poisoning （PSP）. Two sets of four specific primers targeting these two species were derived from the sequence of internal transcribed spacer （ITS） of ribosomal DNA. The method worked well in less than an hour under isothermal conditions of 65℃. LAMP specificity was validated in closely related algae as a comparison, suggesting the strict specificity of the LAMP primers. Two visual inspection approaches were feasible to interpret the positive or negative results. The detection lim- its of A. catenella and A. minutum samples using the LAMP assay were found to be 5.6 and 4.5 pg DNA, respectively. The sensitivity of this LAMP assay was 10 or 100-fold higher than Polymerase Chain Reaction （PCR） method in detecting the two microalgae. These characteristics of species specificity, sensitivity, and rapidity suggest that this method has the potentiality in the monitoring of red tide caused by A. catenella and A. minutum.
文摘Enhanced UV-B(280-320 nm) radiation resulting from ozone depletion is one of global environmental problems. Not only marine organisms but also marine ecosystems can be affected by enhanced UV-B radiation. The effects of UV-B radiation on interaction of macro-algae and micro-algae were investigated using Ulva pertusa Kjellman and Alexandrium tamarense as the materials in this study. The results demonstrated that UV-B radiation could inhibit the growth of Ulva pertusa and Alexandrium tamarense when they were both mono-cultured, and the growth inhibition of algae was more significant with increasing doses of UV-B radiation. Alexandrium tamarense could inhibit the growth of Ulva pertusa in mixed culture, and the growth inhibition was more significant when increasing the initial cell density. However, Ulva pertusa could inhibit the growth of Alexandrium tamarense in early phase and stimulate the growth in latter phase when they were grown in mixed culture. Lower initial cell density(10~2 cell/ml) of Alexandrium tamarense could inhibit the growth of Ulva pertusa under UV-B radiation treatment,however, with the initial cell density increasing(10~3 and 10~4 cell/ml), the growth of Ulva pertusa was stimulated under lower dose of UV-B radiation and inhibited under higher dose of UV-B radiation by Alexandrium tamarense. Compared with that in mixed culture, Ulva pertusa showed more positive inhibition to the growth of Alexandrium tamarense under UV-B radiation treatment.
文摘Paralytic shellfish poisoning (PSP) toxins are potent neurotoxins mainly produced by dinoflagellates and being concentrated in bivalves through food web transfer. Increasing number of findings of toxin-producing bacteria in the cells of dinoflagellate such as Alexandriumtamarense supports the hypothesis of the bacterial origin of PSP toxins. Evidence that there are specific symbiosis bacterial taxa associated with the phytoplankton indicates the presence of specific selective mechanisms between them, and implies that the symbiosis bacteria have some vital function to the benefit of the dinoflagellates. Studies on the role of toxin-producing symbiosis bacteria in the marine ecosystem are considered to be becoming more important. Although toxigenic bacteria could be isolated from toxic dinoflagellates, it was not clearly proven whether the isolated bacterial strains based on culture-dependent manner and the corresponding intracellular bacteria were the same because of microbial unculturability. This paper aims to demonstrate the biodiversity of the symbiotic bacteria associated with toxic dinoflagellate A. tamarense using the culture-indepen- dent high-throughput pyrosequencing method, as well as the phylogenetic analysis based on 16S rDNA sequences of the symbiotic cultivable bacteria strains isolated from toxic Alexander tamarense.
基金This study was supported by the key project of the National 10th Five-Year Plan Programthe research project title was research and development of red tide microalgal toxin (2001BA707B03)
文摘Objective Impact of the presence of bacteria associated with a marine dinoflagellate, Alexandrium tamarense CI01, on the growth and toxin production of the algae in batch culture was investigated. Methods Pronounced changes in the activities of the algal culture were observed when the culture was treated with different doses of a mixture of penicillin and streptomycin. Results In the presence of antibiotics at the initial concentration of 100 u/mL in culture medium, both algal growth and toxin yield increased markedly. When the concentration of antibiotics was increased to 500 u/mL, the microalgal growth was inhibited, but resumed in a few days to eventually reach the same level of growth and toxin production as at the lower dose of the antibiotics. When the antibiotics were present at a concentration of 1 000 u/mL, the algal growth was inhibited permanently. Conclusions The results indicate that antibiotics can enhance algal growth and toxin production not only through their inhibition of the growth and hence competition for nutrients, but also through their effects on the physiology of the algae.
基金This study was supported by the National Basic Research Project under contract No.2001CB409709,National Natural Science Foundation of China under contract Nos 49906007,39950001,20177023 and 40076030,CAS Innovation Program under contract No.KZCX2-206,Dire
文摘The effects of a PSP producing dinoflagellate Alexandrium tamarense on marine bivalvesat their several important life stages: egg, D- shape larva, eyespot larva, juvenile and adult, were studied. The results show that the hatching, survival, activity, filtration and growth were adversely affected by the alga and the impact was significantly increased with the increase of algal density. The inhibitory effect on egg hatching was most significant, which the hatching rate was only 30% of the control when exposed to the alga at 100 cell/cm3 after 36 h. Further experiments show that the algal culture, re-suspended cells and cell fragments had the inhibitory effect, while no such effect was from the cell-free medium, cell contents and standard STX. The results indicate that the alga could produce unknown toxins, rather than PSP, associated with the cell surface.
基金The National Natural Sciences Foundation of China(No. 40406003)
文摘The effect of Ca 2+ on the removal of Alexandrium sp. LC3 under HDTMAB stress was investigated. The results showed that the toxic effect of HDTMAB on Alexandrium sp. LC3 was significantly reduced in the presence of Ca 2+, especially under 4 mmol/L of Ca 2+. To understand the underlying mechanism, the SH group and MDA content of the cell membrane and membrane permeability were measured. It was found that the SH content of cell member increased, the MDA content and membrane permeability decreased when Alexandrium sp. was treated with Ca 2+ and HDTMAB complex, compared with using HDTMAB only. The data suggested that Ca 2+ might promote HDTMAB stress resistance of Alexandrium sp. LC3 by reducing the permeability and increasing the stability of cell membrane.
基金The Nature Science Foundation of Qingdao, China under contract No. 05-1-JC-87International Foundation for Science under contract No.AA/16180 awarded to Sui Zhenghong
文摘Harmful algal blooms （HABs） have been affecting negatively the shellfish and aquaculture industries around the world. Though a lot of efforts have been made to disclose the changes of environmental factors involved and their effects on the HABs events, the molecular mechanism of this process remains unclear. To address this problem, proliferating cell nuclear antigen gene （pcna） was isolated and characterized from Alexandrium catenella. It showed high homology to those of other dinoflagellates （89% and 91% homology to Pfiesteria piscicid and Pyrocystis lunula, respectively）, and also 42%–43% homology to those of plant and animals. The expression level of pcna revealed by quantitative real time PCR was the lowest at the late lagging cell growth phase, increased to the highest at the late exponential phase, and then decreased at the stationary phase. Though the cell growth rate was also changing, no positive correlation between pcna expression level and cell growth rate was displayed throughout the whole cell growth stages （r 2 =0.024 6）. However, the pcna expression level had the similar trend with the change of cell growth rate throughout the whole growing process, e.g., from increasing at the earlier cell growth stage to decreasing at the following stages, though slightly lagging to the latter.
