Objective To study the transfer of paralytic shellfish toxins （PST） using four simulated marine food chains： dinoflagellate Alexandrium tamarense→Artemia Artemia salina→Mysid shrimp Neomysis awatschensis; A. tama...Objective To study the transfer of paralytic shellfish toxins （PST） using four simulated marine food chains： dinoflagellate Alexandrium tamarense→Artemia Artemia salina→Mysid shrimp Neomysis awatschensis; A. tamarense→N. awatschensis; A. tamarense→A, salina→Perch Lateolabrax japonicus; and A. tamarense→L, japonicus. Methods The ingestion of A. tamarense, a producer of PST, by L. japonicus, N. awatschensis, and A. salina was first confirmed by microscopic observation of A. tamarense cells in the intestine samples of the three different organisms, and by the analysis of Chl.a levels in the samples. Toxin accumulation in L. japonicus and N. awatschensis directly from the feeding on A. tamarense or indirectly through the vector of A. salina was then studied, The toxicity of samples was measured using the AOAC mouse bioassay method, and the toxin content and profile of A. tamarense were analyzed by the HPLC method. Results Both A. salina and N. awatschensis could ingest A. tamarense cells. However, the ingestion capability of A. salina exceeded that of N. awatschensis. After the exposure to the culture of A. tamarense （2 000 cells·mL^-1） for 70 minutes, the content of Chl.a in A. salina and N. awatschensis reached 0.87 and 0.024 μg.mg^-1, respectively. Besides, A. tamarense cells existed in the intestines of L. japonicus, N. awatschensis and A. salina by microscopic observation. Therefore, the three organisms could ingest A. tamarense cells directly. A. salina could accumulate high content of PST, and the toxicity of A. salina in samples collected on days 1, 4, and 5 of the experiment was 2.18, 2.6, and 2.1 MU.g^-1, respectively. All extracts from the samples could lead to death of tested mice within 7 minutes, and the toxin content in anemia sample collected on the 1st day was estimated to be 1.65×10 ^5 μg STX equal/individual. Toxin accumulation in L japonicus and N. awatschensis directly from the feeding on A. tamarense or indirectly from the vector ofA. salina was also studied. The mice injected with extracts from L japonicus and N. awatschensis samples that accumulated PST either directly or indirectly showed PST intoxication symptoms, indicating that low levels of PST existed in these samples. Conclusion Paralytic shellfish toxins can be transferred to L. japonicus, N. awatschensis, and A. salina from A. tamarense directly or indirectly via the food chains.展开更多
Sulfotransferase （ST） is the first enzyme discovered in association with paralytic shellfish poisoning （PSP） toxin biosynthesis in toxic dinoflagellates. This study investigates the ST activity m crude enzyme extr...Sulfotransferase （ST） is the first enzyme discovered in association with paralytic shellfish poisoning （PSP） toxin biosynthesis in toxic dinoflagellates. This study investigates the ST activity m crude enzyme extraction of a toxic dinoflagellate species, Alexandrium tamarense CI01. The results show that crude enzyme can transfer a sulfate group from 3＇-phosphoadenosine 5＇-phosphosulfate （PAPS） to N-21 in the carbamoyl group of gonyautoxin 2/3 （GTX2/3） to produce C 1/C2, but is inactive toward STX to produce GTX5. The crude enzyme is optimally active at pH 6.0 and 15℃. The activity is enhanced by Co^2＋, Mg^2＋, Mn^2＋ and Ca^2＋ individually, but is inhibited by Cu^2＋. Moreover, the activity shows no difference when various sulfur compounds are used as sulfate donors. These results demonstrate that the ST specific to GTX2/3 is present in the cells of A. tamarense CI01 and is involved in PSP toxin biosynthesis. In addition, the ST from different dinoflagellates is species-specific, which explains well the various biosynthesis pathways of the PSP toxins in toxic dinoflagellates.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
Dinoflagellate Alexandrium catenella is a cosmopolitan bloom-forming species with complex life cycle,the formation and germination of resting cysts are critical for its bloom dynamics.In the coastal waters of Qinhuang...Dinoflagellate Alexandrium catenella is a cosmopolitan bloom-forming species with complex life cycle,the formation and germination of resting cysts are critical for its bloom dynamics.In the coastal waters of Qinhuangdao,A.catenella has been identified as the major causative agent for paralytic shellfish poisoning,but there is little knowledge concerning its resting cysts in this region.In this study,three surveys were carried out along the coast of Qinhuangdao from 2020 to 2021 to map the distribution of A.catenella resting cysts,using a quantitative PCR(qPCR)assay specific for A.catenella.The resting cysts were detected in surface sediments during all the three surveys,and their distribution patterns were similar.High abundance of resting cysts(maximum 1300 cysts/g sediment(wet weight))were found in a region(119.62°E-119.99°E,39.67°N-39.98°N)northeast to the coastal waters of Qinhuangdao,where surface sediments were mainly composed of clay and silt(percentage above 50%).Prior to the formation of the A.catenella bloom in March 2021,the abundance of A.catenella vegetative cells in seawater had extremely significant positive correlation with the abundance of resting cysts in surface sediments,reflecting the important role of resting cysts in the initiation of A.catenella blooms.As far as we know,this is the first report on the distribution of A.catenella cysts along the coast of Qinhuangdao.The results will of fer a sound basis for the future monitoring and mitigation of toxic A.catenella blooms and paralytic shellfish poisoning events in this region.展开更多
基金The work was supported by National Basic Research Project No. 2001 CB409700, NNSFC KZCX2-YW-208.
文摘Objective To study the transfer of paralytic shellfish toxins （PST） using four simulated marine food chains： dinoflagellate Alexandrium tamarense→Artemia Artemia salina→Mysid shrimp Neomysis awatschensis; A. tamarense→N. awatschensis; A. tamarense→A, salina→Perch Lateolabrax japonicus; and A. tamarense→L, japonicus. Methods The ingestion of A. tamarense, a producer of PST, by L. japonicus, N. awatschensis, and A. salina was first confirmed by microscopic observation of A. tamarense cells in the intestine samples of the three different organisms, and by the analysis of Chl.a levels in the samples. Toxin accumulation in L. japonicus and N. awatschensis directly from the feeding on A. tamarense or indirectly through the vector of A. salina was then studied, The toxicity of samples was measured using the AOAC mouse bioassay method, and the toxin content and profile of A. tamarense were analyzed by the HPLC method. Results Both A. salina and N. awatschensis could ingest A. tamarense cells. However, the ingestion capability of A. salina exceeded that of N. awatschensis. After the exposure to the culture of A. tamarense （2 000 cells·mL^-1） for 70 minutes, the content of Chl.a in A. salina and N. awatschensis reached 0.87 and 0.024 μg.mg^-1, respectively. Besides, A. tamarense cells existed in the intestines of L. japonicus, N. awatschensis and A. salina by microscopic observation. Therefore, the three organisms could ingest A. tamarense cells directly. A. salina could accumulate high content of PST, and the toxicity of A. salina in samples collected on days 1, 4, and 5 of the experiment was 2.18, 2.6, and 2.1 MU.g^-1, respectively. All extracts from the samples could lead to death of tested mice within 7 minutes, and the toxin content in anemia sample collected on the 1st day was estimated to be 1.65×10 ^5 μg STX equal/individual. Toxin accumulation in L japonicus and N. awatschensis directly from the feeding on A. tamarense or indirectly from the vector ofA. salina was also studied. The mice injected with extracts from L japonicus and N. awatschensis samples that accumulated PST either directly or indirectly showed PST intoxication symptoms, indicating that low levels of PST existed in these samples. Conclusion Paralytic shellfish toxins can be transferred to L. japonicus, N. awatschensis, and A. salina from A. tamarense directly or indirectly via the food chains.
基金the National Natural Science Foundation of China (No.40376032)the Ministry of Science and Technology of the People’s Republic of China (No.2001CB409700)
文摘Sulfotransferase （ST） is the first enzyme discovered in association with paralytic shellfish poisoning （PSP） toxin biosynthesis in toxic dinoflagellates. This study investigates the ST activity m crude enzyme extraction of a toxic dinoflagellate species, Alexandrium tamarense CI01. The results show that crude enzyme can transfer a sulfate group from 3＇-phosphoadenosine 5＇-phosphosulfate （PAPS） to N-21 in the carbamoyl group of gonyautoxin 2/3 （GTX2/3） to produce C 1/C2, but is inactive toward STX to produce GTX5. The crude enzyme is optimally active at pH 6.0 and 15℃. The activity is enhanced by Co^2＋, Mg^2＋, Mn^2＋ and Ca^2＋ individually, but is inhibited by Cu^2＋. Moreover, the activity shows no difference when various sulfur compounds are used as sulfate donors. These results demonstrate that the ST specific to GTX2/3 is present in the cells of A. tamarense CI01 and is involved in PSP toxin biosynthesis. In addition, the ST from different dinoflagellates is species-specific, which explains well the various biosynthesis pathways of the PSP toxins in toxic dinoflagellates.
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Supported by the National S&T Basic Resources Investigation Program of China(No.2018YF100206)the National Key R&D Program of China(Nos.2019YFC1407901,2017YFC1600701)from the Ministry of Science and Technology(MoST)。
文摘Dinoflagellate Alexandrium catenella is a cosmopolitan bloom-forming species with complex life cycle,the formation and germination of resting cysts are critical for its bloom dynamics.In the coastal waters of Qinhuangdao,A.catenella has been identified as the major causative agent for paralytic shellfish poisoning,but there is little knowledge concerning its resting cysts in this region.In this study,three surveys were carried out along the coast of Qinhuangdao from 2020 to 2021 to map the distribution of A.catenella resting cysts,using a quantitative PCR(qPCR)assay specific for A.catenella.The resting cysts were detected in surface sediments during all the three surveys,and their distribution patterns were similar.High abundance of resting cysts(maximum 1300 cysts/g sediment(wet weight))were found in a region(119.62°E-119.99°E,39.67°N-39.98°N)northeast to the coastal waters of Qinhuangdao,where surface sediments were mainly composed of clay and silt(percentage above 50%).Prior to the formation of the A.catenella bloom in March 2021,the abundance of A.catenella vegetative cells in seawater had extremely significant positive correlation with the abundance of resting cysts in surface sediments,reflecting the important role of resting cysts in the initiation of A.catenella blooms.As far as we know,this is the first report on the distribution of A.catenella cysts along the coast of Qinhuangdao.The results will of fer a sound basis for the future monitoring and mitigation of toxic A.catenella blooms and paralytic shellfish poisoning events in this region.
