Harmful Algal Bloom

有害的藻华
  • 文章类型: Journal Article
    尽管微藻通常在海洋食物网中充当水母ephyrae的猎物,这项研究调查了有害微藻对月球水母Aureliaaurita产生有害影响的潜力。了解Aurelia和微藻物种之间的生物相互作用至关重要,特别是考虑到它们在全球沿海水域的常见共现。我们检查了11种原生生物菌株的影响,包括七种有害微藻和两种无毒微藻,在A.auritaephyrae上。当暴露于radheterosigmaakashiwo和Chattonellamarinavar时,Aurita的节律脉动行为受到明显抑制。卵黄和鞭毛藻两栖动物,Cooliacanariensis,和PiscicidaPfiesteria.值得注意的是,所有H.akashiwo菌株和C.marinavar的培养基滤液。ovata杀死了Ephyrae,暗示化学物质可能在细胞外释放。这项研究发现了微藻和水母ephyrae之间的新相互作用,暗示有害的藻华可能会抑制水母的大量发生。
    Although microalgae typically serve as prey for jellyfish ephyrae in marine food webs, this study investigated the potential of harmful microalgae to produce detrimental effects on the moon jellyfish Aurelia aurita. Understanding the biological interactions between Aurelia and microalgal species is crucial, particularly considering their common co-occurrence in coastal waters worldwide. We examined the effects of 11 protist strains, comprising seven species of harmful microalgae and two non-toxic microalgae, on A. aurita ephyrae. The rhythmic pulsation behavior of A. aurita was significantly suppressed when exposed to the raphidophytes Heterosigma akashiwo and Chattonella marina var. ovata and the dinoflagellates Amphidinium carterae, Coolia canariensis, and Pfiesteria piscicida. Notably, the media filtrates of all H. akashiwo strains and C. marina var. ovata killed ephyrae, implying a possible extracellular release of chemicals. This study discovered novel interactions between microalgae and jellyfish ephyrae, implying that harmful algal blooms may suppress mass occurrences of Aurelia medusae.
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  • 文章类型: Journal Article
    尽管在伊比利亚半岛海岸报告的第一次赤潮是由于多林格洛丁,了解他们的频率和,特别是,关于促成开花的环境条件仍然很少。出于这个原因,根据1993-2008年记录数据库期间,在夏季的三个加利西亚rias中观察和研究了L.polyedra开花事件;此外,样本于2008年夏季收集。6月和8月,在阿瑞斯和巴奎罗的rias中发生了Polyedra的增殖,分别,在科鲁尼亚的Ria,他们从六月底坚持到九月初。当地表温度达到17℃时,出现了赤潮,具有“季节性热窗”条件,当盐度≥30时,即,“最佳盐度窗口”;当这些参数低于这些阈值时,囊肿萌发减少。还必须存在从沉积物到表面的囊肿运输机制;在Barqueiro的ria中发现这种机制是自然的(潮流),或者在Ares和Coruña的rias中是人为的(疏浚)。在1993-2008年期间,夏季的表面温度通常有利于囊肿萌发(85%至100%);但是,低于10m深度的水温很少达到17°C的阈值(2%至18%)。在这16年期间,疏浚活动可以解释记录的开花事件的71%(Coruña)和44%(Ares)。当盛开在初夏时,有利条件没有导致新的赤潮,可能是由于囊肿发芽所需的滞后期。此外,由于特定年份的夏季上升流脉冲,当水柱中仍有高密度的硅藻(>1,000,000个细胞·L-1)时,不会出现水华。在这项研究中发现的温度-沉积物干扰模式为预防这种鞭毛藻赤潮导致的最终风险提供了有用的工具。
    Despite the fact that the first red tide reported on the coasts of the Iberian Peninsula was due to Lingulodinium polyedra, knowledge about their frequency and, particularly, about the environmental conditions contributing to bloom initiation is still scarce. For this reason, L. polyedra bloom episodes were observed and studied in three Galician rias during the summer season based on the 1993-2008 record database period; additionally, samples were collected in summer 2008. Proliferations of L. polyedra occurred in the rias of Ares and Barqueiro in June and August, respectively, while in the Ria of Coruña, they persisted from the end of June to early September. Red tides developed when the surface temperature reached 17 °C, with \"seasonal thermal window\" conditions, and when salinities were ≥30, i.e., an \"optimal salinity window\"; when these parameters were lower than these thresholds, cyst germination decreased. A cyst transport mechanism from sediments to the surface must also exist; this mechanism was found to be natural (tidal currents) in the ria of Barqueiro or anthropogenic (dredging) in the rias of Ares and Coruña. Surface temperatures during summer were usually favorable for cyst germination (85 to 100%) during the 1993-2008 period; however, water temperatures below 10 m depth only rarely reached the 17 °C threshold (2 to 18%). During this 16-year period, dredging activities could explain 71% (Coruña) and 44% (Ares) of the recorded bloom events. When a bloom episode developed in early summer, favorable conditions did not lead to a new red tide, probably due to the lag period required by cysts for germination. Moreover, blooms did not develop when high densities of diatoms (>1,000,000 cells·L-1) remained in the water column as a result of summer upwelling pulses occurring in specific years. The temperature-sediment disturbance pattern found in this study provides a useful tool for the prevention of eventual risks resulting from red tides of this dinoflagellate.
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  • 文章类型: Journal Article
    有害藻华(HAB)的形成导致水生态系统的富营养化,并可能使休闲湖泊不适合人类使用。我们评估了元编码的适用性和比较,宏基因组学,qPCR,以及基于ELISA的方法,用于检测大湖地区的水华和非水华地区的蓝细菌/蓝毒素。基于DNA测序的方法强有力地识别了布卢姆和非布卢姆样品之间的差异(例如,鱼腥草和浮游植物的相对突出)。Shotgun测序策略还确定了水华样品中典型的蓝藻代谢基因的富集,尽管没有检测到毒素基因,提示可能需要更深入的测序或PCR方法来检测低丰度的毒素基因。PCR和ELISA表明,盛开部位的微囊藻毒素水平和微囊藻毒素基因拷贝明显更丰富。然而,并非所有的水华样本都对微囊藻毒素呈阳性,可能是由于不产生毒素的物种的开花发展。此外,微囊藻毒素水平与微囊藻毒素基因拷贝数显着相关(正相关),但与蓝藻16S基因总拷贝数无关。总之,基于下一代测序的方法可以识别特定的分类和功能靶标,可用于绝对定量方法(qPCR和ELISA)以增强常规的水监测策略。
    Harmful algal bloom (HAB) formation leads to the eutrophication of water ecosystems and may render recreational lakes unsuitable for human use. We evaluated the applicability and comparison of metabarcoding, metagenomics, qPCR, and ELISA-based methods for cyanobacteria/cyanotoxin detection in bloom and non-bloom sites for the Great Lakes region. DNA sequencing-based methods robustly identified differences between bloom and non-bloom samples (e.g., the relative prominence of Anabaena and Planktothrix). Shotgun sequencing strategies also identified the enrichment of metabolic genes typical of cyanobacteria in bloom samples, though toxin genes were not detected, suggesting deeper sequencing or PCR methods may be needed to detect low-abundance toxin genes. PCR and ELISA indicated microcystin levels and microcystin gene copies were significantly more abundant in bloom sites. However, not all bloom samples were positive for microcystin, possibly due to bloom development by non-toxin-producing species. Additionally, microcystin levels were significantly correlated (positively) with microcystin gene copy number but not with total cyanobacterial 16S gene copies. In summary, next-generation sequencing-based methods can identify specific taxonomic and functional targets, which can be used for absolute quantification methods (qPCR and ELISA) to augment conventional water monitoring strategies.
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  • 文章类型: Journal Article
    鱼毒素赤潮是世界面临和需要解决的问题。使用来自海洋大型藻类的抗藻化合物来抑制鱼毒素赤潮被认为是一种有前途的生物防治方法。从Bangiafusco-purpurea中筛选并分离出抗藻类物质,Gelidiumamansii,Gloiopeltis分叉,Hizikiafusifarme,海带,PalmariaPalmata,和Sargassumsp.利用生物活性引导的分离方法,获得开发抗鱼毒性赤潮微藻除藻剂的新材料。7种大型藻类的馏分对两栖动物和卡列尼亚克莫托表现出选择性抑制活性,其中乙酸乙酯组分对两种测试的赤潮微藻具有最强和最广泛的抗藻活性。它们对A.carterae和K.mikimootoi的抑制作用甚至强于重铬酸钾,如紫菜芽孢杆菌的乙酸乙酯馏分,H.fusifarme,和Sargassumsp.进一步进行薄层色谱和紫外光谱以筛选Sargassumsp的乙酸乙酯级分。最后,一种新的糖脂衍生物,2-O-二十烷酰基-3-O-(6-氨基-6-脱氧)-β-D-吡喃葡萄糖基-甘油,是从马尾藻中分离和鉴定的。,它是第一次从海洋大型藻类中分离出来。确定了2-O-二十烷酰基-3-O-(6-氨基-6-脱氧)-β-D-吡喃葡萄糖基甘油对A.carterae和K.mikimootoi的显着抗藻作用。
    Ichthyotoxic red tide is a problem that the world is facing and needs to solve. The use of antialgal compounds from marine macroalgae to suppress ichthyotoxic red tide is considered a promising biological control method. Antialgal substances were screened and isolated from Bangia fusco-purpurea, Gelidium amansii, Gloiopeltis furcate, Hizikia fusifarme, Laminaria japonica, Palmaria palmata, and Sargassum sp. to obtain new materials for the development of algaecides against ichthyotoxic red tide microalgae using bioactivity-guided isolation methods. The fractions of seven macroalgae exhibited selective inhibitory activities against Amphidinium carterae and Karenia mikimotoi, of which the ethyl acetate fractions had the strongest and broadest antialgal activities for the two tested red tide microalgae. Their inhibitory effects on A. carterae and K. mikimotoi were even stronger than that of potassium dichromate, such as ethyl acetate fractions of B. purpurea, H. fusifarme, and Sargassum sp. Thin-layer chromatography and ultraviolet spectroscopy were further carried out to screen the ethyl acetate fraction of Sargassum sp. Finally, a new glycolipid derivative, 2-O-eicosanoyl-3-O-(6-amino-6-deoxy)-β-D-glucopyranosyl-glycerol, was isolated and identified from Sargassum sp., and it was isolated for the first time from marine macroalgae. The significant antialgal effects of 2-O-eicosanoyl-3-O-(6-amino-6-deoxy)-β-D-glucopyranosyl-glycerol on A. carterae and K. mikimotoi were determined.
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  • 文章类型: Journal Article
    逻辑方程以sigmoid曲线对单个物种的种群增长进行建模,该曲线以指数形式开始,以渐近方法结束,以自然系统承载能力确定的最终种群。但是自然系统的人口在接近承载能力时通常不会稳定。相反,它表现出周期性的变化,有时振幅非常大。逻辑方程的时间延迟修改通过将当前的人口增长率与较早的条件联系起来来解释这种行为。人口随时间的周期性变化可以从单调方法发展到承载能力;到围绕承载能力的振荡;限制循环周期性变化;和,最后,混乱的变化。多个物种的存在以及采样频率和空间覆盖不足阻碍了时间延迟逻辑方程在现实世界种群中的应用。沿着西南佛罗里达墨西哥湾沿岸的卡列尼亚短命绽放,然而,提供了一个独特的机会,因为花朵几乎是单一的,并且在广泛的地理区域频繁采样,他们是测试时滞逻辑方程的好候选人。我们表明,这些水华的浓度峰值在40-100天的范围内,与时滞Logistic方程预测的结果一致。峰之间的谷中的细胞浓度比峰值低至少2-3个数量级,为潜在的缓解努力提供可预测的机会窗口。
    The logistic equation models single-species population growth with a sigmoid curve that begins as exponential and ends with an asymptotic approach to a final population determined by natural system carrying capacity. But the population of a natural system often does not stabilize as it approaches carrying capacity. Instead, it exhibits periodic change, sometimes with very large amplitudes. The time-delay modification of the logistic equation accounts for this behavior by connecting the present rate of population growth to conditions at an earlier time. The periodic change in population with time can progress from a monotonic approach to the carrying capacity; to oscillation around the carrying capacity; to limit-cycle periodic change; and, finally, to chaotic change. The presence of multiple species and inadequate sampling frequency and spatial coverage hinder the application of the time-delay logistic equation to real-world populations. Blooms of Karenia brevis along the southwest Florida Gulf Coast, however, provide a unique opportunity in that blooms are nearly monospecific and are sampled frequently over a wide geographic region; they are good candidates for testing the time-delay logistic equation. We show that these blooms exhibit peaks in concentration with periods in the range of 40-100 days, consistent with that predicted by the time-delay logistic equation. Cell concentrations in the valleys between the peaks are at least 2-3 orders of magnitude lower than peak values, offering predictable windows of opportunity for potential mitigation efforts.
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  • 文章类型: Journal Article
    由于各种水体中的有毒蓝藻水华在周围区域释放有害物质,因此如今受到了广泛关注。虾池塘中的这些有毒浮游蓝细菌极大地影响了虾的存活。厄瓜多尔是仅次于巴西的美洲第二大虾生产国;由于厄瓜多尔虾池中有毒的蓝藻水华,以虾为基础的经济受到威胁。这项研究调查了厄瓜多尔太平洋沿岸的Chone和Jama河(位于Manabi省)的虾池中不同蓝细菌的丰度,关注不同的环境因素,如温度,pH值,盐度,和光。温度和pH是影响蓝藻丰度的关键因素,raphiopsisraciborskii与pH之间呈显着正相关。在干旱季节,在Chone和Jama河附近的虾池中发现的蓝细菌丰度最高和最低,分别为>3×106和1×106Cell。分别为m-3。Chone和Jama河虾池的Shannon-Wiener多样性指数在0.41-1.15和0.31-1.15之间波动,分别。这种变化与盐度的变化和有害藻华的存在有关,强调持续监测的重要性。此外,研究区域显示出多样性低的富营养化条件,强调需要在两条河流中进行额外的时空研究和扩大研究,更好地理解这些复杂的现象。这些发现强调了持续监测和扩大蓝藻生态学研究的重要性,对公共卫生和水生资源管理有影响。
    Toxic cyanobacterial blooms in various water bodies have been given much attention nowadays as they release hazardous substances in the surrounding areas. These toxic planktonic cyanobacteria in shrimp ponds greatly affect the survival of shrimps. Ecuador is the second highest shrimp producing country in the Americas after Brazil; and the shrimp-based economy is under threat due to toxic cyanobacterial blooms in Ecuador shrimp ponds. This study investigated the abundance of different cyanobacteria in the shrimp ponds at the Chone and Jama rivers (in Manabi province) at Ecuadorian pacific coast, focusing on different environmental factors, such as temperature, pH, salinity, and light. Temperature and pH were identified as key factors in influencing the abundance of cyanobacteria, with a significant positive correlation between Raphidiopsis raciborskii and pH. The highest and lowest abundance of cyanobacteria found during the dry season in the shrimp ponds near the Chone and Jama rivers were > 3 × 106 and 1 × 106 Cell.m-3, respectively. The Shannon-Wiener Diversity Index fluctuated between 0.41-1.15 and 0.31-1.15 for shrimp ponds of Chone and Jama rivers, respectively. This variation was linked to changes in salinity and the presence of harmful algal blooms, highlighting the importance of continuous monitoring. Additionally, the study areas showed eutrophic conditions with low diversity, underlining the need for additional spatiotemporal studies and expanded research in both rivers, to better understand these complex phenomena. The findings underscore the importance of continuous monitoring and expanded research in cyanobacteria ecology, with implications for public health and aquatic resource management.
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  • 文章类型: Journal Article
    垂直迁移行为,这是海洋能源循环不可或缺的一部分,是海洋生物中普遍存在的特征。然而,浮游植物的行为,特别是在diel垂直迁移(DVM)之外,与浮游动物等群体相比,仍未充分开发。通过有害藻类Heterosigmaakashiwo的镜头,表现出独特的垂直迁移和波动的赤潮模式,本研究旨在探索浮游植物垂直迁移行为的生态复杂性和多样性益处。在H.akashiwo的盛开时期,我们意外地在白天观察到底层密集的细胞浓度。这一阶段与该物种有性生殖相关细胞的出现相吻合。实验室实验进一步表明,与地表相比,迁移到更深深度的细胞群中有性生殖的频率更高。这一发现暗示了密集的底部积累(BA)与物种的生命周期转变之间的联系。这个BA阶段持续了两天,之后,人口恢复到标准的DVM行为,深入了解赤石独特的红潮波动模式。我们的研究表明,浮游植物的垂直迁移并不是由DVM严格决定的,揭示了不同的垂直迁移行为,这些行为可能导致有害藻华模式的复杂性。
    Vertical migration behaviour, which is integral to marine energy circulation, is a prevalent trait among marine organisms. However, the behaviour of phytoplankton, particularly beyond diel vertical migration (DVM), remain underexplored compared to groups like zooplankton. Through the lens of the harmful alga Heterosigma akashiwo, which exhibits unique vertical migrations and fluctuating red tide patterns, this study aimed to explore the ecological intricacies and diverse benefits of phytoplankton vertical migration behaviours. During the bloom period of H. akashiwo, we unexpectedly observed a dense concentration of cells at bottom layer during daytime. This phase coincided with the emergence of cells related to this species\' sexual reproduction. Laboratory experiments further showed an elevated frequency of sexual reproduction in the cell populations that migrated to deeper depths compared to those at the surface. This finding implies a connection between dense bottom accumulation (BA) and the life cycle transitions of the species. This BA phase persisted for two days, after which the populations returned to their standard DVM behaviour, providing insight into the unique fluctuating red tide patterns of H. akashiwo. Our study suggests that phytoplankton vertical migrations are not strictly dictated by DVM, revealing diverse vertical migration behaviours that may contribute to the complexity of harmful algal bloom patterns.
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  • 文章类型: Journal Article
    由于废水处理不足和农业养分输入过多,Winam海湾(肯尼亚)经常受到蓝细菌有害藻华(cHAB)的损害。虽然维多利亚湖的浮游植物已经使用形态学标准进行了表征,我们的目标是使用分子方法鉴定潜在的产毒蓝细菌.连续两个夏季对海湾进行了采样,进行16S和18S核糖体RNA基因测序。此外,通过定量PCR检查了参与氰基毒素产生的关键基因。细菌群落是空间可变的,形成与海湾地区一致的不同集群。与重氮营养相关的类群在霍马湾附近占主导地位。在东边,样本显示出cyrA丰度升高,表明圆柱精子合成的遗传能力。的确,2022年,在Nyando河口附近,CyrA超过1000万份L-1,其中有6000多个Cylindrospermopsis。细胞mL-1。相比之下,西南地区在Homa湾附近检测到mcyE基因(微囊藻毒素合成)升高,微囊藻和Dolichospermumspp。被观察到。这些发现表明,在相对较小的范围内,cHABs的组成和毒素合成潜力可以显著变化。这突出表明需要采取多方面的管理方法和经常监测蓝藻毒素,以减少对人类健康的影响。
    The Winam Gulf (Kenya) is frequently impaired by cyanobacterial harmful algal blooms (cHABs) due to inadequate wastewater treatment and excess agricultural nutrient input. While phytoplankton in Lake Victoria have been characterized using morphological criteria, our aim is to identify potential toxin-producing cyanobacteria using molecular approaches. The Gulf was sampled over two successive summer seasons, and 16S and 18S ribosomal RNA gene sequencing was performed. Additionally, key genes involved in production of cyanotoxins were examined by quantitative PCR. Bacterial communities were spatially variable, forming distinct clusters in line with regions of the Gulf. Taxa associated with diazotrophy were dominant near Homa Bay. On the eastern side, samples exhibited elevated cyrA abundances, indicating genetic capability of cylindrospermopsin synthesis. Indeed, near the Nyando River mouth in 2022, cyrA exceeded 10 million copies L-1 where there were more than 6000 Cylindrospermopsis spp. cells mL-1. In contrast, the southwestern region had elevated mcyE gene (microcystin synthesis) detections near Homa Bay where Microcystis and Dolichospermum spp. were observed. These findings show that within a relatively small embayment, composition and toxin synthesis potential of cHABs can vary dramatically. This underscores the need for multifaceted management approaches and frequent cyanotoxin monitoring to reduce human health impacts.
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  • 文章类型: Journal Article
    全球分布的有害藻华(HAB)物种,Heterosigmaakashiwo,已经发现表现出鱼鳞病毒性。先前的研究表明,H.akashiwo通过抑制共存硅藻的生长,在开花期间获得了竞争优势,Skeletonemacostatum,通过化感作用。然而,赤蜡对S.costatum的具体化感作用机制尚不清楚。为了弥合这个差距,本研究采用实时定量PCR和代谢组学相结合的方法,研究了赤石草对S.costatum的化感过程.我们的结果表明,与H.akashiwo共培养时,S.costatum的生长受到阻碍(初始细胞浓度,2×104细胞/mL)。基因表达调查显示细胞色素b6,核糖二磷酸羧化酶大链的mRNA水平显着降低,当在共培养条件下生长时,S.costatum中的硅转运蛋白。此外,代谢途径分析表明,赤石草的化感作用破坏了S.costatum的几个重要代谢途径,包括嘌呤和嘧啶代谢的减少和脂肪酸生物合成的增加。我们的调查揭示了化感作用在H.akashiwo花的形成中的复杂而实质性的参与,证明了akashiwo和S.costatum之间化感作用的复杂性。这些见解也大大有助于我们对HAB物种内部动力学的理解。
    The globally distributed harmful algal blooms (HAB) species, Heterosigma akashiwo, has been found to exhibit ichthyotoxicity. Previous studies have shown that H. akashiwo achieves a competitive edge during bloom occurrences by inhibiting the growth of a coexisting diatom, Skeletonema costatum, through allelopathy. However, the specific allelopathic mechanisms underlying the allelopathic effects of H. akashiwo on S. costatum remain unknown. To bridge this gap, our study utilized a combination of quantitative real-time PCR and metabolomics to examine the allelopathic processes of H. akashiwo on S. costatum. Our results demonstrate that the growth of S. costatum is hindered when co-cultured with H. akashiwo (initial cell concentration, 2 × 104 cell/mL). Gene expression investigation showed a substantial reduction in the mRNA levels of cytochrome b6, ribulose bisphosphate carboxylase large chain, and silicon transporter in S. costatum when grown in co-culture conditions. Furthermore, metabolic pathway analysis suggested that the allelopathic effects of H. akashiwo disrupted several vital metabolic pathways in S. costatum, including a reduction in purine and pyrimidine metabolism and an increase in fatty acid biosynthesis. Our investigation has revealed the intricate and substantial involvement of allelopathy in the formation of H. akashiwo blooms, demonstrating the complexity of the allelopathic interaction between H. akashiwo and S. costatum. These insights also contribute significantly to our understanding of the dynamics within HAB species.
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  • 文章类型: Journal Article
    背景:有害藻华(HAB),由微生物的快速增殖或聚集引起,对环境来说是灾难性的。Prymnesiumparvum是一种在世界范围内发现的藻类物种,负责幼虫两栖动物和双壳类动物的大量繁殖和死亡,对生态环境造成严重的负面影响。为了防止和管理环境污染,使用简单的方法探索和制定HABs的现场早期检测策略至关重要。与早期检测相关的主要挑战是准确和灵敏地检测低丰度的藻类。
    结果:这里,重组酶聚合酶扩增(RPA)与成簇的规则间隔短回文重复序列和Cas12a蛋白(CRISPR-LbaCas12a)系统相结合,首次使用侧流试纸(LFD)对小疟原虫进行早期检测。选择细小疟原虫的内部转录间隔区(ITS)作为靶序列,以及单链DNA报告基因的浓度,缓冲液系统,反应时间,优化了金颗粒的用量。RPA-CRISPR-LbaCas12a-LFD方法在实验测试过程中表现出高度特异性,对用作对照的不同微藻没有交叉反应。此外,最低检测限比独立RPA方法的最低检测限好10,000倍.利用不同的环境样本进一步验证了该方法的可行性和鲁棒性。它还观察到细小疟原虫在中国海域广泛分布,但细小疟原虫的细胞密度相对较低(<0.1个细胞/mL)。
    结论:开发的方法具有出色的特异性,并且比独立的RPA方法具有10,000倍的灵敏度。这些优点使这种方法适用于环境水中HAB事件的预警检测和预防。此外,这项研究的结果可以促进从传统的实验室检测到现场监测的转变,促进对HAB的早期预警。
    BACKGROUND: Harmful algal blooms (HABs), caused by the rapid proliferation or aggregation of microorganisms, are catastrophic for the environment. The Prymnesium parvum is a haptophyte algal species that is found worldwide and is responsible for extensive blooms and death of larval amphibians and bivalves, causing serious negative impacts on the ecological environment. For the prevention and management of environmental pollution, it is crucial to explore and develop early detection strategies for HABs on-site using simple methods. The major challenge related to early detection is the accurate and sensitive detection of algae present in low abundance.
    RESULTS: Herein, recombinase polymerase amplification (RPA) was combined with clustered regularly interspaced short palindromic repeats and Cas12a protein (CRISPR-LbaCas12a) systems, and the lateral flow dipstick (LFD) was used for the first time for early detection of P. parvum. The internal transcribed spacer (ITS) of P. parvum was selected as the target sequence, and the concentration of single-strand DNA reporters, buffer liquid system, reaction time, and amount of gold particles were optimized. The RPA-CRISPR-LbaCas12a-LFD approach demonstrated highly specificity during experimental testing, with no cross-reaction against different microalgae used as controls. In addition, the lowest detection limit was 10,000 times better than the lowest detection limit of the standalone RPA approach. The feasibility and robustness of this approach were further verified by using the different environmental samples. It also observed that P. parvum are widely distributed in Chinese Sea, but the cell density of P. parvum is relatively low (<0.1 cells/mL).
    CONCLUSIONS: The developed approach has an excellent specificity and offers 10,000 times better sensitivity than the standalone RPA approach. These advantages make this approach suitable for early warning detection and prevention of HAB events in environmental water. Also, the outcomes of this study could promote a shift from traditional laboratory-based detection to on-site monitoring, facilitating early warning against HABs.
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