Indole Alkaloids of the Stigonematales (Cyanophyta): Chemical Diversity,Biosynthesis and Biological Activity

The cyanobacteria are well recognized as producers of a wide array of bioactive metabolites including toxins, and potential drug candidates. However, a limited number of taxa are generally considered with respect to both of these aspects. That said, the order Stigonematales, although largely overlooked in this regard, has become increasingly recognized as a source of bioactive metabolites relevant to both human and environmental health. In particular, the hapalindoles and related indole alkaloids (i.e., ambiguines, fischerindoles, welwitindolinones) from the order, represent a diverse, and phylogenetically characteristic, class of secondary metabolites with biological activity suggestive of potential as both environmental toxins, and promising drug discovery leads. The present review gives an overview of the chemical diversity of biologically active metabolites from the Stigonematales—and particularly the so-called hapalindole-type alkaloids—including their biosynthetic origins, and their pharmacologically and toxicologically relevant bioactivities. Taken together, the current evidence suggests that these alkaloids, and the associated cyanobacterial taxa from the order, warrant future consideration as both potentially harmful (i.e., “toxic”) algae, and as promising leads for drug discovery.     

Lipophilic Toxins in Chile: History,Producers and Impacts

A variety of microalgal species produce lipophilic toxins (LT) that are accumulated by filter-feeding bivalves. Their negative impacts on human health and shellfish exploitation are determined by toxic potential of the local strains and toxin biotransformations by exploited bivalve species. Chile has become, in a decade, the world’s major exporter of mussels (Mytilus chilensis) and scallops (Argopecten purpuratus) and has implemented toxin testing according to importing countries’ demands. Species of the Dinophysis acuminata complex and Protoceratium reticulatum are the most widespread and abundant LT producers in Chile. Dominant D. acuminata strains, notwithstanding, unlike most strains in Europe rich in okadaic acid (OA), produce only pectenotoxins, with no impact on human health. Dinophysis acuta, suspected to be the main cause of diarrhetic shellfish poisoning outbreaks, is found in the two southernmost regions of Chile, and has apparently shifted poleward. Mouse bioassay (MBA) is the official method to control shellfish safety for the national market. Positive results from mouse tests to mixtures of toxins and other compounds only toxic by intraperitoneal injection, including already deregulated toxins (PTXs), force unnecessary harvesting bans, and hinder progress in the identification of emerging toxins. Here, 50 years of LST events in Chile, and current knowledge of their sources, accumulation and effects, are reviewed. Improvements of monitoring practices are suggested, and strategies to face new challenges and answer the main questions are proposed.     

Beta-N-methylamino-l-alanine: LC-MS/MS Optimization,Screening of Cyanobacterial Strains and Occurrence in Shellfish from Thau,a French Mediterranean Lagoon

β-N-methylamino-l-alanine (BMAA) is a neurotoxic non-protein amino acid suggested to be involved in neurodegenerative diseases. It was reported to be produced by cyanobacteria, but also found in edible aquatic organisms, thus raising concern of a widespread human exposure. However, the chemical analysis of BMAA and its isomers are controversial, mainly due to the lack of selectivity of the analytical methods. Using factorial design, we have optimized the chromatographic separation of underivatized analogues by a hydrophilic interaction chromatography coupled to tandem mass spectrometry (HILIC-MS/MS) method. A combination of an effective solid phase extraction (SPE) clean-up, appropriate chromatographic resolution and the use of specific mass spectral transitions allowed for the development of a highly selective and sensitive analytical procedure to identify and quantify BMAA and its isomers (in both free and total form) in cyanobacteria and mollusk matrices (LOQ of 0.225 and 0.15 µg/g dry weight, respectively). Ten species of cyanobacteria (six are reported to be BMAA producers) were screened with this method, and neither free nor bound BMAA could be found, while both free and bound DAB were present in almost all samples. Mussels and oysters collected in 2009 in the Thau Lagoon, France, were also screened, and bound BMAA and its two isomers, DAB and AEG, were observed in all samples (from 0.6 to 14.4 µg/g DW), while only several samples contained quantifiable free BMAA.     

茶叶氟研究进展:累积特性、含量及安全性评价

综述了茶园土壤氟、茶树氟吸收累积特性及茶叶氟安全性评价等方面的主要研究进展,包括土壤氟含量水平及变化规律、茶树品种间氟吸收差异、pH、铝、钙和环境条件等对茶树氟吸收的的影响、主要茶类氟含量现状、释放态势及与人体健康关系等方面。     

Evolution and Distribution of Saxitoxin Biosynthesis in Dinoflagellates

Numerous species of marine dinoflagellates synthesize the potent environmental neurotoxic alkaloid, saxitoxin, the agent of the human illness, paralytic shellfish poisoning. In addition, certain freshwater species of cyanobacteria also synthesize the same toxic compound, with the biosynthetic pathway and genes responsible being recently reported. Three theories have been postulated to explain the origin of saxitoxin in dinoflagellates: The production of saxitoxin by co-cultured bacteria rather than the dinoflagellates themselves, convergent evolution within both dinoflagellates and bacteria and horizontal gene transfer between dinoflagellates and bacteria. The discovery of cyanobacterial saxitoxin homologs in dinoflagellates has enabled us for the first time to evaluate these theories. Here, we review the distribution of saxitoxin within the dinoflagellates and our knowledge of its genetic basis to determine the likely evolutionary origins of this potent neurotoxin.     

Diarrhetic Shellfish Toxins and Other Lipophilic Toxins of Human Health Concern in Washington State

The illness of three people in 2011 after their ingestion of mussels collected from Sequim Bay State Park, Washington State, USA, demonstrated the need to monitor diarrhetic shellfish toxins (DSTs) in Washington State for the protection of human health. Following these cases of diarrhetic shellfish poisoning, monitoring for DSTs in Washington State became formalized in 2012, guided by routine monitoring of Dinophysis species by the SoundToxins program in Puget Sound and the Olympic Region Harmful Algal Bloom (ORHAB) partnership on the outer Washington State coast. Here we show that the DSTs at concentrations above the guidance level of 16 μg okadaic acid (OA) + dinophysistoxins (DTXs)/100 g shellfish tissue were widespread in sentinel mussels throughout Puget Sound in summer 2012 and included harvest closures of California mussel, varnish clam, manila clam and Pacific oyster. Concentrations of toxins in Pacific oyster and manila clam were often at least half those measured in blue mussels at the same site. The primary toxin isomer in shellfish and plankton samples was dinophysistoxin-1 (DTX-1) with D. acuminata as the primary Dinophysis species. Other lipophilic toxins in shellfish were pectenotoxin-2 (PTX-2) and yessotoxin (YTX) with azaspiracid-2 (AZA-2) also measured in phytoplankton samples. Okadaic acid, azaspiracid-1 (AZA-1) and azaspiracid-3 (AZA-3) were all below the levels of detection by liquid chromatography tandem mass spectrometry (LC-MS/MS). A shellfish closure at Ruby Beach, Washington, was the first ever noted on the Washington State Pacific coast due to DSTs. The greater than average Fraser River flow during the summers of 2011 and 2012 may have provided an environment conducive to dinoflagellates and played a role in the prevalence of toxigenic Dinophysis in Puget Sound.     

Characterization of intracellular and extracellular saxitoxin levels in both field and cultured Alexandrium spp. samples from Sequim Bay, Washington

Traditionally, harmful algal bloom studies have primarily focused on quantifying toxin levels contained within the phytoplankton cells of interest. In the case of paralytic shellfish poisoning toxins (PSTs), intracellular toxin levels and the effects of dietary consumption of toxic cells by planktivores have been well documented. However, little information is available regarding the levels of extracellular PSTs that may leak or be released into seawater from toxic cells during blooms. In order to fully evaluate the risks of harmful algal bloom toxins in the marine food web, it is necessary to understand all potential routes of exposure. In the present study, extracellular and intracellular PST levels were measured in field seawater samples (collected weekly from June to October 2004–2007) and in Alexandrium spp. culture samples isolated from Sequim Bay, Washington. Measurable levels of intra- and extra-cellular toxins were detected in both field and culture samples via receptor binding assay (RBA) and an enzyme-linked immunosorbent assay (ELISA). Characterization of the PST toxin profile in the Sequim Bay isolates by pre-column oxidation and HPLC-fluorescence detection revealed that gonyautoxin 1 and 4 made up 65 ± 9.7 % of the total PSTs present. Collectively, these data confirm that extracellular PSTs are present during blooms of Alexandrium spp. in the Sequim Bay region.     

三种农药在茶汤中的浸出及对人体健康影响的风险评价

选择三唑磷、硫丹、氯氟氰菊酯三种农药,在生长均匀的茶园喷施,采摘施药后第2、5、9、14天和21天的茶树鲜叶加工成绿茶,获得含不同浓度农药残留的成茶样品,用气相色谱法测定了农药在成茶、茶汤和茶渣中的残留含量动态,研究表明：成茶和茶汤中的残留水平随施药间隔天数的增加呈逐渐下降趋势,茶汤中农药残留的浓度与成茶中的浓度呈正相关且为线性关系,平均浸出率分别为29.06％(三唑磷)、5.11％(硫丹)、1.73％(氯氟氰菊酯),不溶于茶汤的农药残留大部分存留于茶渣中。将人体因饮茶而摄入的农药残留量与农药ADI相比较,结果表明饮茶摄入的此三种农药的残留对人体健康影响的风险非常小,仅在10^-3~10^-5 水平上。     

Genetic and Geographic Patterns of Duplicate DPL Genes Causing Genetic Incompatibility Within Rice: Implications for Multiple Domestication Events in Rice

Strong genetic incompatibilities exist between two primary rice subspecies,indica and japonica.However,the wild ancestors of rice,O.nivara Sharma et Shastry and O.rufipogon Griff.,are genetically compatible.How this genetic incompatibility became established has not been clearly elucidated.To provide insights into the process,we analyzed a pair of hybrid sterility genes in rice,DOPPELGANGER 1(DPL1)and DOPPELGANGER 2(DPL2).Either of the two loci can have one defective allele(DPL1-and DPL2-).Hybrid pollen carrying both DPL1-and DPL2-alleles is sterile.To explore the origination of DPL1-and DPL2-,we sequenced the DPL1 and DPL2 genes of 811 individual plants,including Oryza sativa(132),O.nivara(296)and O.rufipogon(383).We then obtained 20 DPL1 and 34 DPL2 sequences of O.sativa from online databases.Using these sequences,we analyzed the genetic and geographic distribution patterns of DPL genes in modern rice and its wild ancestors.Compared with the ancestral populations,DPL1-and DPL2-showed reduced diversity but increased frequency in modern rice.We speculated that the diversity reduction was due to a historic genetic bottleneck,and the frequency had likely increased because the defective alleles were preferred following this artificial selection.Such results indicated that standing variances in ancestral lines can lead to severe incompatibilities among descendants.Haplotype analysis indicated that the DPL1-haplotype of rice emerged from an O.nivara population in India,whereas the DPL2-haplotype emerged from O.rufipogon in South China.Hence,the evolutionary history of DPLs conforms to the presumed multiple domestication events of modern rice.     

防治水稻白背飞虱高毒农药替代药剂的室内筛选及对吡虫啉的抗性风险评估

 为筛选高毒农药的替代药剂,采用稻茎浸渍法测定了7类共21种杀虫剂对2006年和2007年采自南京市江浦地区的白背飞虱种群3龄若虫的毒力。结果表明,噻虫嗪、吡虫啉、噻嗪酮对白背飞虱3龄若虫的毒力最高(LC50=004~026 mg/L);烯啶虫胺、丁烯氟虫腈、吡蚜酮、啶虫咪、氯噻啉、毒死蜱、氟虫腈等也有较高的毒力(LC50＜4.50 mg/L）;异丙威、敌敌畏虽然触杀毒力稍低(LC50＞19.85 mg/L),但两者在生产上仍广泛应用,所以把以上12种药剂作为田间药效试验的推荐品种;同时还比较了该地区白背飞虱种群对供试药剂敏感性的年度间差异,其中对毒死蜱、异丙威、丁硫克百威、速灭威这4种药剂的敏感性下降了66.7%~71.4%。对2006年南京江浦种群用吡虫啉连续筛选15代的结果表明,1~15代抗性上升433倍,抗性现实遗传力为0.2295,存在抗性风险。另外,还讨论了白背飞虱的综合防治。     

Implications of climate change for grassland in Europe: impacts, adaptations and mitigation options: a review

Climate change associated with greenhouse gas (GHG) emissions may have important implications for Europe's grasslands. Projected scenarios indicate that increased temperatures and CO₂ concentrations have the potential to increase herbage growth and to favour legumes more than grasses, but changes in seasonal precipitation would reduce these benefits particularly in areas with low summer rainfall. Further implications for grasslands may arise from increased frequency of droughts, storms and other extreme events. Potential farm-scale adaptive responses to climate change are identified. Grassland agriculture also contributes to GHG emissions, particularly methane and nitrous oxide, and management of grassland affects net carbon balances and carbon sequestration. Management options are identified for mitigating grassland's contribution to GHG emissions which need to be developed in a holistic way that also considers other pressures.