Harmful cyanobacteria and their cyanotoxins in Egyptian fresh waters – state of knowledge and research needs

Cyanobacterial blooms have increased in freshwater ecosystems worldwide in the last century, mostly resulting from eutrophication and climate change. These blooms represent serious threats to environmental and human health because of the production of harmful metabolites, called cyanotoxins. Like many countries, Egypt has been plagued with cyanobacterial blooms in most water sources, including the Nile River, irrigation canals, lakes and fishponds. However, the data about cyanotoxins produced in these blooms are limited. Only two types of cyanotoxins, microcystins and cylindrospermopsin, have been identified and characterised, mainly from Microcystis and Cylindrospermopsis blooms. The data revealed the presence of microcystins in raw and treated drinking waters at concentrations (0.05–3.8 µg l^?1), exceeding the WHO limit (1 µg l^?1) in some drinking water treatment plants. In addition, Nile tilapia Oreochromis niloticus caught from ponds containing heavy cyanobacterial blooms have accumulated considerable amounts of cyanotoxins in their edible tissues. The data presented here could be the catalyst for the establishment of a monitoring and management programme for harmful cyanobacteria and their cyanotoxins in Egyptian fresh waters. This review also elucidates the important research gaps and possible avenues for future research on cyanobacterial blooms and cyanotoxins in Egypt.     

水华蓝藻对鱼类的营养毒理学效应

水体富营养化导致蓝藻水华的发生已成为全球关注的水环境问题,很多鱼类处于水生态系统食物链的最高级,蓝藻水华的主要次级代谢产物-微囊藻毒素可通过鱼类的摄食活动或生物富集作用在鱼体组织中累积,并通过食物链危及人类健康。近年来,微囊藻毒素对鱼类的毒性效应引起众多科学家的关注。在天然水体中不少鱼类可以主动摄食蓝藻,所以,水华蓝藻对鱼类来说既具有营养物作用、也具有潜在的毒性作用。鉴于目前机械收获的水华蓝藻生物量资源化利用问题以及水产饲料业亟需大力开发鱼粉替代蛋白源的需要,从营养学和毒理学这两个角度来研究水华蓝藻对鱼类的营养作用和毒性效应具有较高的理论和现实意义。主要概述了蓝藻粉、蓝藻细胞对鱼类的营养学和毒理学效应,以期拓展水华蓝藻对鱼类毒性效应的研究视野,同时也为水华蓝藻的资源化利用提供新的思路。     

A 4700-Year History of Cyanobacteria Toxin Production in a Shallow Subtropical Lake

Cyanobacterial toxins or cyanotoxins are secondary metabolites produced by cyanobacteria and are found in water bodies around the world causing negative effects on aquatic ecosystems and human health. There are numerous environmental and biological triggers for toxin production, and the ecological role of most toxins is still being determined. Whereas cyanobacterial toxin occurrence appears to be expanding and monitoring efforts have increased in recent years, the history of toxin existence in lakes is poorly understood. Here, I report the history of the cyanotoxin, cylindrospermopsin (CYN), in sediments of hypereutrophic Lake Griffin, Florida, USA, from approximately 4700 years ago to present. The record includes three periods of toxin abundance: one associated with recent, European settlement in the watershed, and the other two during the middle to late Holocene, prior to human impacts on the lake. Each period corresponds to changes in different paleolimnological measurements suggesting drivers of CYN production have varied through time. This CYN record demonstrates the use of sediment toxin concentrations as a tool to reconstruct historic cyanobacterial toxin occurrence and shows that toxin production can occur independent of anthropogenic stressors.     

Cyanotoxin production and phylogeny of benthic cyanobacterial strains isolated from the northeast of Brazil

Most of the knowledge about cyanobacteria toxin production is traditionally associated with planktonic cyanobacterial blooms. However, some studies have been showing that benthic cyanobacteria can produce cyanotoxins. According to this, we aimed to evaluate the production of microcystins and saxitoxins in benthic cyanobacteria isolated from aquatic ecosystems in the Northeast of Brazil and to use a polyphasic approach for their identification. Forty-five clonal strains were isolated from rivers and water supply reservoirs, and identified using morphological and molecular phylogenetic characteristics. In order to evaluate the toxins production, the strains were screened for genes involved in the biosynthesis of microcystins and saxitoxins, positive results were confirmed and cyanotoxins quantified using HPLC. Eight species were identified belonging to the Phormidiaceae, Pseudanabaenaceae and Nostocaceae families. This is the first study in Brazil that shows that strains from the Geitlerinema genus correspond to at least three phylogenetic lineages, which possibly correspond to three distinct species to be subsequently reclassified. The strains that showed one of the genes involved in the cyanotoxins production were analyzed by HPLC and Geitlerinema amphibium, Geitlerinema lemmermannii, Cylindrospermum stagnale and Phormidium uncinatum were identified as producing one or more saxitoxins variants. Thus, this is the first report of saxitoxins production for those first three species and the first report in Brazil for P. uncinatum.     

Health Effects of Toxin-Producing Cyanobacteria: “The CyanoHABs”

Increasingly, harmful algal blooms (HABs) are being reported worldwide due to several factors, primarily eutrophication, climate change and more scientific monitoring. All but cyanobacteria toxin poisonings (CTPs) are mainly a marine occurrence. CTPs occur in fresh (lakes, ponds, rivers and reservoirs) and brackish (seas, estuaries, and lakes) waters throughout the world. Organisms responsible include an estimated 40 genera but the main ones are Anabaena, Aphanizomenon, Cylindrospermopsis, Lyngbya, Microcystis, Nostoc, and Oscillatoria (Planktothrix). Cyanobacteria toxins (cyanotoxins) include cytotoxins and biotoxins with biotoxins being responsible for acute lethal, acute, chronic and sub-chronic poisonings of wild/domestic animals and humans. The biotoxins include the neurotoxins; ana-toxin-a, anatoxin-a(s) and saxitoxins plus the hepatotoxins; microcystins, nodularins and cylindrospermopsins. Confirmations of human deaths from cyanotoxins are limited to exposure through renal dialysis at a haemodialysis center in Caruaru, Brazil, in 1996. A major effort to compile all available information on toxic cyanobacteria including issues of human health, safe water practices, management, prevention and remediation have been published by the World Health Organization. This paper will review our current understanding of CTP's including their risk to human health.     

Use of qPCR for the study of hepatotoxic cyanobacteria population dynamics

Toxic cyanobacteria blooms are increasingly frequent and object of greater concern due to its ecological and health impacts. One important lack in the toxic cyanobacteria research field is to understand which parameters influence most and how they operate to regulate the overall levels of cyanotoxins in a body of water. MC concentration is believed to be influenced by changes in several seasonal environmental factors that influence the succession of toxic cyanobacteria. In the last years, qPCR (quantitative polymerase chain reaction) has been applied to determine the seasonal and temporal shifts in the proportions of MC-producing and non-MC-producing subpopulations by quantifying both mcy genotypes and total population numbers. We discuss the most prominent and recent studies using qPCR to address hepatotoxic cyanobacteria population dynamics and evaluate how they helped understanding the factors promoting the growth of toxic strains in situ and the succession of hepatotoxin-producing genera in natural populations.     

Dark side of cyanobacteria: searching for strategies to control blooms

Cyanobacteria are ecologically one of the most prolific groups of photosynthetic prokaryotes in marine and freshwater habitats. They are primary producer microorganisms and are involved in the production of important secondary metabolites, including toxic compounds such as cyanotoxins. Environmental conditions promote massive growth of these microbes, causing blooms that can have critical ecological and public health implications. In this highlight, we discuss some of the approaches being addressed to prevent these blooms, such as control of nutrient loading, treatments to minimize growth or monitoring interactions with other species.     

蓝藻毒素对底栖动物的毒理学研究进展

近年,由于人类活动加剧,大量氮磷等营养物质流入湖泊等缓流水体,导致水体富营养化。而由此引起有害蓝藻水华的频繁爆发,使生态环境和人类健康受到严重威胁。相关研究表明,蓝藻水华的爆发不仅能够使水体水质恶化,其中一些产毒藻类还会产生大量蓝藻毒素,对水生生物产生重要影响。底栖动物作为水体生态系统的重要组成部分,在食物网中有重要作用,同时其中的许多种类又与人类息息相关,因此关于水华蓝藻毒素对淡水底栖动物的毒理学研究具有重要意义。在介绍蓝藻毒素概况的基础上,综述了蓝藻毒素的致毒机理和对底栖动物的影响,展望了研究方向。     

Evaluation of sampling and screening techniques for tiered monitoring of toxic cyanobacteria in lakes

Exposure to cyanotoxins can pose serious human health consequences both through drinking water and recreational use of a contaminated waterway. We adapted a tiered framework proposed by the World Health Organization (WHO) for monitoring cyanobacteria populations and assessing public health risks and implemented the program in Lake Champlain. This study focused on evaluating the sampling protocols employed in this adapted monitoring program over two field seasons (2003 and 2004). Using a paired sampling design, we evaluated whether 63-μm Wisconsin net samples adequately represented whole-water conditions and whether chlorophyll a concentration could serve as a useful predictor of cyanobacteria density and microcystin concentration. We also evaluated the spatial and temporal dynamics of blooms and their implications for monitoring. Our results suggest that using threshold values of either potentially toxic cyanobacteria density counted using a rapid screening protocol or chlorophyll a concentration serve as initial indicators of potentially high levels of cyanotoxin; however, the utility of chlorophyll a is system-dependent. Whole-water samples provide more accurate estimates of population density and higher microcystin concentrations than net samples, offering a more cautionary approach to assessing risk to recreational lake users. Shoreline samples generally showed higher cyanotoxin concentrations than offshore, but restricting sampling to the shoreline may miss early warnings of bloom development.     

Effects of Cyanobacteria on Survival and Reproduction of the Littoral Crustacean Gammarus zaddachi (Amphipoda)

Toxic cyanobacteria can have harmful or fatal impacts on aquatic organisms. In the archipelagos of the northern Baltic Sea, the open sea blooms often drift into littoral areas, where they decompose and release toxins and other chemical compounds in the water. However, the effects of cyanobacteria on the littoral organisms have not previously been investigated. We studied the effects of three cyanobacteria species (toxic Nodularia spumigena, non-toxic N. sphaerocarpa and non-toxic Aphanizomenon flos-aquae) and purified dissolved nodularin (produced by N. spumigena) on a common littoral amphipod Gammarus zaddachi. Nodularin was transferred to eggs, juveniles and adults of G. zaddachi, but no significant negative effects of dissolved nodularin were detected on adults, eggs or juveniles. However, survival of adults decreased by the exposure to toxic N. spumigena cells. The egg hatching rate and juvenile survival were not affected when exposed to the three cyanobacteria species. In contrast, a weak decrease in the egg production and an increased abortion of embryos from the brood pouch of females was observed, the later indicating a failure in parental care. Further, a decrease in grazing rate on the filamentous green alga Enteromorpha intestinalis was observed. The results suggest that toxic cyanobacteria blooms are not extremely fatal, but may have, in high concentrations, negative effects on the adult survival, fecundity, and feeding behaviour of gammarids inhabiting the littoral zone.     

Environmental factors associated with toxic cyanobacterial blooms across 20 drinking water reservoirs in a semi-arid region of Brazil

Cyanobacteria are known to produce a wide variety of bioactive, toxic secondary metabolites generally described as hepatotoxins, neurotoxins, cytotoxins, or dermatoxins. In Brazil, the regular monitoring of cyanobacterial toxins has intensified after the death of 65 patients in a hemodialysis clinic in Caruaru in the state of Pernambuco due to microcystin exposure. The primary objective of this study was to use multivariate statistics that incorporated environmental parameters (both biotic and abiotic) to forecast blooms of cyanobacteria and their toxic secondary metabolites in 20 drinking water reservoirs managed by the Water Treatment Company of Ceará (CAGECE) in the semi-arid region of Ceará, Brazil. Across four years (January 2013 to January 2017), 114 different phytoplankton taxa were identified, including 24 cyanobacterial taxa. In general, Ceará reservoirs were dominated by cyanobacteria due to eutrophication but also because of the dry and warm climate found throughout the region. Interestingly, specific cyanobacterial taxa were influenced by different biotic and abiotic factors. For example, nitrogen-to-phosphorus (N:P) and evaporation were positively related to saxitoxin-producing taxa, especially Raphidiopsis raciborskii, while temperature, electrical conductivity, total phosphorus, and transparency (measured as Secchi depth) were positively associated with microcystin-producing taxa, such as Microcystis aeruginosa. Climate forecasts predict higher evaporation and temperatures in the semi-arid Ceará region, which will likely magnify droughts and water scarcity as well as promote toxic cyanobacterial blooms in reservoirs in the future. Therefore, understanding the factors associated with algal blooms dominated by specific taxa is paramount for water resource management.     

Estimating toxic cyanobacteria in a Brazilian reservoir by quantitative real-time PCR, based on the microcystin synthetase D gene

The production of microcystin toxins by cyanobacteria is an intrapopulation feature and the toxic and nontoxic genotypes can be separated only through molecular analyses targeting the mcy markers. Quantitative real-time PCR (qPCR) is a procedure that has been established, not only to detect but to specifically quantify these genotypes. In the present work, primers were designed for the mcyD region to estimate the number of cyanobacteria that are potential microcystin producers. Laboratory tests to verify the efficiency and the specificity of the primers were performed. The methodology was first established for single strain cultures and thereafter was applied in environmental water samples, from a reservoir located in the Brazilian savannah (“cerrado”). The results were very satisfactory, demonstrating the high efficiency and the specificity of the primers used, and their ability to detect different cyanobacteria genera. Of particular interest were the results showing a high proportion of toxic strains (as high as 100 %) in the environmental samples, as previously reported in another tropical system. Furthermore, the occurrence of a smaller fraction of toxic strains at high cyanobacteria densities, and of more toxic populations when fewer cyanobacteria were present, deserves further investigation. Although records of cyanobacteria blooms are very common in the tropics and suggest an increasing incidence of toxic populations, the present research is one of the few applying qPCR in a tropical environment. The results obtained here, by a technique that allows a more precise quantification and in situ follow-up of changes in toxicity, will make possible new observations of seasonal and spatial dynamics in these environments.     

应用环境微生物治理淡水湖泊微囊藻毒素污染的研究进展

近年来,随着水体富营养化程度的加剧,蓝藻水华现象时有发生,蓝藻及其释放的藻毒素对生态环境和人类健康构成严重威胁。在各种藻毒素中,以微囊藻毒素(Microcystins,MCs)毒性最强,对人类危害也最大,微囊藻毒素的化学性质相对稳定且难以通过常规水处理方法消除,因此如何有效去除环境中的MCs是国内外普遍关注的难题。研究发现自然界中的微生物能够有效降解和消除MCs污染,由此产生的环保技术极具应用价值。本文主要概述了微囊藻毒素的产生机理、化学结构以及毒性危害,总结了微囊藻毒素的自然分解过程以及微生物群落对微囊藻毒素的响应机制,重点分析了微生物群落在微囊藻毒素污染控制技术中的潜在应用,并对应用微生物技术治理微囊藻毒素污染的技术瓶颈提出了建议,以期加速微囊藻毒素微生物降解技术的完善和应用。     

PRESENCE AND DIVERSITY OF ALGAL TOXINS IN SUBTROPICAL PEATLAND PERIPHYTON: THE FLORIDA EVERGLADES,USA1

Production of toxic secondary metabolites by cyanobacteria, collectively referred to as cyanotoxins, has been well described for eutrophied water bodies around the world. However, cohesive cyanobacterial mats also comprise a significant amount of biomass in subtropical oligotrophic wetlands. As these habitats generally do not support much secondary production, cyanotoxins, coupled with other physiological attributes of cyanobacteria, may be contributing to the minimized consumer biomass. Periphyton from the Florida Everglades has a diverse and abundant cyanobacterial assemblage whose species produce toxic metabolites; therefore, by screening periphyton representative of the greater Everglades ecosystem, six different cyanotoxins and one toxin (domoic acid) produced by diatoms were identified, ranging in content from 3 × 10^?9 to 1.3 × 10^?6 (g · g^?1), with saxitoxin, microcystin, and anatoxin‐a being the most common. While content of toxins were generally low, when coupled with the tremendous periphyton biomass (3–3,000 g · m^?2), a significant amount of cyanotoxins may be present. While the direct effects of the toxins identified here on the local grazing community need to be determined, the screening process utilized proved effective in showing the broad potential of periphyton to produce a variety of toxins.     

Current approaches to cyanotoxin risk assessment and risk management around the globe

Toxic cyanobacteria became more widely recognized as a potential health hazard in the 1990s, and in 1998 the World Health Organization (WHO) first published a provisional Guideline Value of 1 μg L⁻¹ for microcystin-LR in drinking-water. In this publication we compare risk assessment and risk management of toxic cyanobacteria in 17 countries across all five continents. We focus on the three main (oral) exposure vehicles to cyanotoxins: drinking-water, water related recreational and freshwater seafood. Most countries have implemented the provisional WHO Guideline Value, some as legally binding standard, to ensure the distribution of safe drinking-water with respect to microcystins. Regulation, however, also needs to address the possible presence of a wide range of other cyanotoxins and bioactive compounds, for which no guideline values can be derived due to insufficient toxicological data. The presence of microcystins (commonly expressed as microcystin-LR equivalents) may be used as proxy for overall guidance on risk management, but this simplification may miss certain risks, for instance from dissolved fractions of cylindrospermopsin and cyanobacterial neurotoxins. An alternative approach, often taken for risk assessment and management in recreational waters, is to regulate cyanobacterial presence – as cell numbers or biomass – rather than individual toxins. Here, many countries have implemented a two or three tier alert level system with incremental severity. These systems define the levels where responses are switched from Surveillance to Alert and finally to Action Mode and they specify the short-term actions that follow. Surface bloom formation is commonly judged to be a significant risk because of the elevated concentration of microcystins in a scum. Countries have based their derivations of legally binding standards, guideline values, maximally allowed concentrations (or limits named otherwise) on very similar scientific methodology, but underlying assumptions such as bloom duration, average body size and the amount of water consumed while swimming vary according to local circumstances. Furthermore, for toxins with incomplete toxicological data elements of expert judgment become more relevant and this also leads to a larger degree of variation between countries’ thresholds triggering certain actions. Cyanobacterial blooms and their cyanotoxin content are a highly variable phenomenon, largely depending on local conditions, and likely concentrations can be assessed and managed best if the specific conditions of the locality are known and their impact on bloom occurrence are understood. Risk Management Frameworks, such as for example the Water Safety Plan concept of the WHO and the ‘bathing water profile’ of the European Union are suggested to be effective approaches for preventing human exposure by managing toxic cyanobacteria from catchment to consumer for drinking water and at recreational sites.     

The cyanotoxin-microcystins: current overview

The monocyclic heptapeptides microcystins (MCs), are a group of hepatotoxins, produced worldwide by some bloom-forming cyanobacterial species/strains both in marine and freshwater ecosystems. MCs are synthesized non-ribosomally by large multi-enzyme complexes consisting of different modules including polyketide synthases and non-ribosomal peptide synthetases, as well as several tailoring enzymes. More than 85 different variants of MCs have been reported to exist in nature. These are chemically stable, but undergo bio-degradation in natural water reservoirs. Direct or indirect intake of MCs through the food web is assumed to be a highly exposed route in risk assessment of cyanotoxins. MCs are the most commonly found cyanobacterial toxins that cause a major challenge for the production of safe drinking water and pose a serious threat to global public health as well as fundamental ecological processes due to their potential carcinogenicity. Here, we emphasize recent updates on different modes of action of their possible carcinogenicity. Besides the harmful effects on human and animals, MC producing cyanobacteria can also present a harmful effect on growth and development of agriculturally important plants. Overall, this review emphasizes the current understanding of MCs with their occurrence, geographical distribution, accumulation in the aquatic as well as terrestrial ecosystems, biosynthesis, climate-driven changes in their synthesis, stability and current aspects on its degradation, analysis, mode of action and their ecotoxicological effects.     

运用分子生物学技术监测水环境中产毒蓝藻

有毒蓝藻在形成水华破坏水体环境的同时,也对人畜产生危害。运用分子生物学技术对水环境中产毒蓝藻进行监测,由于其简单、快速和经济等优点,逐渐成为各国学者关注的热点。本文从以下3个方面对其进行了综述:1)早期分子生物学技术在产毒蓝藻诊断中的运用;2)以产毒基因为靶标进行的产毒蓝藻诊断;3)运用基因芯片技术对产毒蓝藻进行检测。与传统的分子生物学技术相比,生物芯片技术具有体积小、重量轻、便于携带、分析自动快速、高通量等许多传统方法所不能比拟的优势。因此,该技术在蓝藻毒素检测中的运用必将给目前的产毒蓝藻的检测带来一场新的革命。     

Consumption of toxic cyanobacteria by Potamopyrgus antipodarum (Gastropoda, Prosobranchia) and consequences on life traits and microcystin accumulation

Among the wide range of toxins produced by cyanobacterial blooms, microcystins (MCs) are the most common and are known to accumulate in aquatic organisms. Freshwater gastropods are grazers and likely to ingest toxic cyanobacteria, particularly Planktothrix agardhii, one of the most common species in the northern hemisphere. The study examines (i) the ingestion of toxic P. agardhii by the prosobranch Potamopyrgus antipodarum, (ii) the kinetics of MC accumulation and depuration in snail tissues during and post-exposure, and (iii) the impact of MCs on their life traits (survival, growth and fecundity). We showed that P. antipodarum ingested 71% of cyanobacteria available during the first 24 h in the presence or not of non-toxic food, and accumulated 1.3% of ingested MCs during the 5-week intoxication period. Elimination of MCs was total after 3 weeks of depuration. A decrease of growth and fecundity was observed during the intoxication period, but it was reversible after the end of exposure. Results are discussed in terms of variation of the response between prosobranch and pulmonate gastropods to toxic cyanobacteria exposure, and the negative impact of toxic cyanobacteria on natural communities of freshwater gastropods.     

Cyanophage and algal virus

<正>Cyanobacteria and eukaryotic algae are abundant in freshwater and marine environments,and are fundamentally important in global biogeochemical cycles and for primary productivity.Under certain conditions,such as an excess of nutrients due to human activities through fertilizers or sewage,