Scientific assessment of freshwater harmful algal blooms

In 2004, Congress reauthorized the Harmful Algal Bloom and Hypoxia Research and Control Act of 1998 with the Harmful Algal Bloom and Hypoxia Amendments Act (HABHRCA 2004). The 2004 legislation required the generation of five reports, including this "Scientific Assessment of Freshwater Harmful Algal Blooms." HABHRCA 2004 stipulates that this report 1) examine the causes, consequences, and economic costs of freshwater HABs, 2) establish priorities and guidelines for a research program on freshwater HABs, and 3) make recommendations to improve coordination among Federal agencies with respect to research on HABs in freshwater environments. This report is divided into five chapters: Chapter 1 provides the legislative background and process for developing the report, Chapter 2 describes the problem of freshwater and inland HABs in the United States, Chapter 3 outlines the current Federal efforts in freshwater and inland HAB research and response, Chapter 4 discusses the future research priorities, and Chapter 5 delineates opportunities for coordination to advance research efforts. The document is based, in large part, on the proceedings (Hudnell 2008) of the International Symposium on Cyanobacterial Harmful Algal Blooms, a meeting convened by EPA and sponsored by a variety of Federal agencies, to describe current scientific knowledge and identify priorities for future research on CyanoHABs. This report offers a plan for coordinating the important research that is currently ongoing in the United States and for guiding future research directions for Federal programs as well as for state, local, private, and academic institutions in order to maximize advancements. To this end, the Interagency Working Group on Harmful Algal Blooms, Hypoxia, and Human Health (IWG-4H) identifies seven priorities, all of equal weight, for freshwater HAB research and response. These priorities represent research areas where there is the greatest potential for progress in freshwater HAB research. This report does not attempt to assess the relative importance of freshwater HAB research compared to other research areas or other priorities for Federal or state investment.

Harmful algal bloom management and response: assessment and plan

This report, "Harmful Algal Bloom Management and Response: Assessment and Plan" reviews and evaluates Harmful Algal Bloom (HAB) management and response efforts, identifies current prevention, control, and mitigation programs for HABs, and presents an innovative research, event response, and infrastructure development plan for advancing the response to HABs. In December 2004, Congress enacted and the President signed into law the Harmful Algal Bloom and Hypoxia Amendments Act of 2004, (HABHRCA 2004). The reauthorization of HABHRCA acknowledged that HABs are one of the most scientifically complex and economically damaging coastal issues challenging our ability to safeguard the health of our Nation’s coastal ecosystems. The Administration further recognized the importance of HABs as a high priority national issue by specifically calling for the implementation of HABHRCA in the President’s U.S. Ocean Action Plan. HABHRCA 2004 requires four reports to assess and recommend research programs on HABs in U.S. waters. This document comprises two linked reports specifically aimed at improving HAB management and response: the Prediction and Response Report and the follow-up plan, the National Scientific Research, Development, Demonstration, and Technology Transfer (RDDTT) Plan on Reducing Impacts from Harmful Algal Blooms. This document was prepared by the Interagency Working Group on Harmful Algal Blooms, Hypoxia, and Human Health, which was chartered through the Joint Subcommittee on Ocean Science and Technology of the National Science and Technology Council and the Interagency Committee on Ocean Science and Resource Management Integration. This report complements and expands upon HAB-related priorities identified in Charting the Course for Ocean Science in the United States for the Next Decade: An Ocean Research Priorities Plan and Implementation Strategy, recently released by the Joint Subcommittee on Ocean Science and Technology. It draws from the contributions of numerous experts and stakeholders from federal, state, and local governments, academia, industry, and non-governmental organizations through direct contributions, previous reports and planning efforts, a public comment period, and a workshop convened to develop strategies for a HAB management and response plan. Given the importance of the Nation’s coastal ocean, estuaries, and inland waters to our quality of life, our culture, and the economy, it is imperative that we move forward to better understand and mitigate the impacts of HABs which threaten all of our coasts and inland waters. This report is an effort to assess the extent of federal, state and local efforts to predict and respond to HAB events and to identify opportunities for charting a way forward.

Biodiversity and production of bioactive material from cyanobacteria in Persian Gulf

In this project sampling was done from 9 stations in 3 depths during 5 seasons to separate, identify and examine the biodiversity of cyanobacteria. Another sampling also has been done to analyse all physical and physicochemical parameters, primary production and polluting agents such as heavy metals and oil hydrocarbons in water and sediments. Along with optimization of Oscillatoria to study the ability of producing natural substances, these cyanobacteria were analysed from the point of antimicrobial and mutagenic effects. To examine the relationships among analysed parameters, the regression test, analysis of varian and Post Hoc were used. As the result of this study 48 genus of cyanobacteria were pecognised among which 35.5% were croco ccales and 64.4% were Hermogonals. Oscillatoria was one of the Filamentous cyanobacteria which had antibacterial and mutagenic effects. The results of multicommunity consistency varians test, post Hoc and homogenous subsets show significant difference between biodiversity of cyanotbacteria in coral ecosystem, Mangrove and transite. The linear correlation coefficient between biodiversity of cyanobacteria and bioenvironmental agents were examined, but there was no continuous relation between these factors and biological biodiversity. In Surface layer there was a significant correlation coefficent at 0.048 and probability at 95% confidence interval. Also, the biodiversity is depended on oil pollution and heavy metals such as copper (Cu) and chromium (Cr).

Identification of triosephosphate isomerase (TIM) genes from Microcystis (Cyanobacteria : Chroococcales) and theoretical analyses of the potential of cyanobacterial TIM as a target for designing specific inhibitors

The genes encoding triosephosphate isomerase (TIM) in three species of Microcystis (M. aeruginosa, M. viridis and M. wesenbergii) were investigated. Reverse transcriptase-polymerase chain reaction indicated that they were transcribed in the cells. Analyses showed that their DNA and deduced amino acid sequences were highly conserved between all the three species, only a single nonsynonymous substitution was seen at position 31, from an Asp in M. aeruginosa and M. viridis to Glu in M. wesenbergii. Sequence alignment of these with 12 other known cyanobacterial TIM sequences showed that all the cyanobacterial TIMs had a very high level of amino acid identity (over 50% between each two). Comparison of the cyanobacterial TIMs with other reported TIMs (from diverse lineages of the three Domains) showed that they possessed common active-site residues and sequence motifs. All cyanobacterial TIMs have two common cysteine residues (Cys127 and Cys176), and the Cys176 is almost cyanobacteria-specific with only one exception in Streptomyces coelicolor. Both secondary structure alignment and comparative modelling of Synechocystis sp. TIM showed that Cys176 was located at the hinge region of the flexible loop-6 and might therefore be critical to the movement of TIM's loop-6, which is important to the function of the enzyme. Thus, the cyanobacterial TIM-specific Cys176 may be a potential site for the discovery of suitable drugs against cyanobacteria, and such drugs may have utility in controlling water blooms due to cyanobacteria.

Consumption of fa cai Nostoc soup: A Potential for BMAA exposure from Nostoc cyanobacteria in China?

Grown in arid regions of western China the cyanobacterium Nostoc flagelliforme - called fa cai in Mandarin and fat choy in Cantonese - is wild-harvested and used to make soup consumed during New Year's celebrations. High prices, up to $125 USD/kg, led to overharvesting in Inner Mongolia, Ningxia, Gansu, Qinghai, and Xinjiang. Degradation of arid ecosystems, desertification, and conflicts between Nostoc harvesters and Mongol herdsman concerned the Chinese environmental authorities, leading to a government ban of Nostoc commerce. This ban stimulated increased marketing of a substitute made from starch. We analysed samples purchased throughout China as well as in Chinese markets in the United States and the United Kingdom. Some were counterfeits consisting of dyed starch noodles. A few samples from California contained Nostoc flagelliforme but were adulterated with starch noodles. Other samples, including those from the United Kingdom, consisted of pure Nostoc flagelliforme. A recent survey of markets in Cheng Du showed no real Nostoc flagelliforme to be marketed. Real and artificial fa cai differ in the presence of beta-N-methylamino-L-alanine (BMAA). Given its status as a high-priced luxury food, the government ban on collection and marketing, and the replacement of real fa cai with starch substitutes consumed only on special occasions, it is anticipated that dietary exposure to BMAA from fa cai will be reduced in the future in China.

Comparative studies on photosynthesis and phosphate metabolism of Cylindrospermopsis raciborskii with Microcystis aeruginosa and Aphanizomenon flos-aquae

The physiological differences for three bloom-forming cyanobacteria (Cylindrospermopsis raciborskii, Microcystis aeruginosa, and Aphanizomenon flos-aquae) were investigated. In comparison with M. aeruginosa and A. flos-aquae, C. raciborskii exhibited a significantly higher concentration of carotenoids, higher values in maximum photosynthesis rate (P-m), apparent photosynthetic efficieny (a), and maximum electron transport rate (ETRmax) during the growth period. In addition, higher extracellular alkaline phosphatase activities and lower light compensation point (I-c) were also detected in C raciborskii (p < 0.05, ANOVA). Therefore, it is suggested that the higher photosynthetic activities, more effective uptake and utilization to phosphate, and low light requirements might play important roles in the occurrence and invasive behavior of C. raciborskii. Crown Copyright (C) 2009 Published by Elsevier B.V. All rights reserved.

Effects of dietary cyanobacteria of two different sources on growth and recovery of hybrid tilapia (Oreochromis niloticus x O. aureus)

A 115 days feeding trial was conducted to evaluate the effect of dietary cyanobacteria on growth, microcystins (MCs) accumulation in hybrid tilapia (Oreochromis niloticus x O. aureus) and the recovery when the fish were free of cyanobacteria. Three experimental diets were formulated: the control (cyanobacteria free diet); one test diet with cyanobacteria from Lake Taihu (AMt 80.0 mu g MCs g(-1) diet) and one with cyanobacteria from Lake Dianchi (AMd, 410.0 rho g MCs g(-1) diet). Each diet was fed to fish for 60 days and then all fish were free of cyanobacteria for another 55 days. A significant increase in feeding rate (FR) was observed in fish fed AMd diet after a first 30-day exposure (1(st) EP), and in fish fed both AMt diet and AMd diet after a second 30-day exposure (2(nd) EP). Specific growth rates (SGR) of fish fed AMt diet and AMd diet were both obviously affected after the first 30-day exposure, but SGR was only significantly affected in fish fed AMt diet after the second 30-day exposure. After a 55-day recovery, there were no significant differences among diets in the indices mentioned above. Much higher concentrations of MCs were accumulated in tissues of all fish exposed to cyanobacteria. After the 55-day recovery, MC concentrations in fish tissues were significantly lower than those on day 60. (C) 2009 Elsevier Ltd. All rights reserved.

Allelopathic mechanism of pyrogallol to Microcystis aeruginosa PCC7806 (Cyanobacteria): From views of gene expression and antioxidant system

Pyrogallol is a potent allelochemical on Microcystis aeruginosa, but its allelopathic mechanism is not fully known. In order to explore this mechanism, gene expressions for prx, mcyB, psbA, recA, grpE, fabZ under pyrogallol stress were studied, and activities of the main antioxidant enzymes were also measured. The results showed that expression of grpE and recA showed no significant change under pyrogallol stress, while psbA and mcyB were up-regulated at 4 mg L-1. Both prx and fabZ were up-regulated even under exposure to 1 mg L-1 pyrogallol concentration. The activities of superoxide dismutase (SOD) and catalase (CAT) were enhanced under pyrogallol stress. Levels of malodialdehyde (MDA) at 2 and 4 mg L-1 pyrogallol were significantly higher than those of the controls. It was concluded that oxidant damage is an important mechanism for the allelopathic effect of pyrogallol on M. aeruginosa. (c) 2009 Elsevier Ltd. All rights reserved.

Allelopathic interactions between the Potamogeton spp and toxic cyanobacteria (Microcystis aeruginosa)

This study aimed to investigate the allelopathic activities between 3 Potamogeton spp. (Potamogeton maackianus, Potamogeton malaianus and Potamogeton pectinatus) and the toxic cyanobacteria (Microcystis aeruginosa). All Potamogeton spp inhibited the growth of M. aeruginosa in both coexistence and exudates experiments. Inhibition of M. aeruginosa growth by plant exudates depended strongly on the biomass of P malaianus. Initial pH (6.5-9.8) did not influence the inhibitory effects of P. malaianus exudates. However, the M. aeruginosa inhibited the net photosynthesis and respiration of all three pondweed test spp.. The decreases in photosynthesis and respiration were probably caused by the toxic compounds released by M. aeruginosa, rather than its shading effects. The M. aeruginosa also decreased the nutrients (phosphorus and nitrogen) uptake rates of macrophytes. The absorption rates of phosphorus and nitrogen and net photosynthesis were decreased sharply. These results will help to restore submerged plants in eutrophic waters.

Effects of Arsenate on Microcystin Content and Leakage of Microcystis Strain PCC7806 Under Various Phosphate Regimes

Both arsenic pollution and eutrophication are prominent environmental issues when considering the problem of global water pollution. It is important to reveal the effects of arsenic species on cyanobacterial growth and toxin yields to assess ecological risk of arsenic pollution or at least understand naturally occurring blooms. The sensitivity of cyanobacteria to arsenate has often been linked to the structural similarities of arsenate and phosphate. Thus, we approached the effect of arsenate with concentrations from 10(-8) to 10(-4) M on Microcystis strain PCC7806 under various phosphate regimes. The present study showed that Microcystis strain PCC7806 was arsenate tolerant up to 10(-4) M. And such tolerance was without reference to both content of intra- and extra-cellular phosphate. It seems that arsenate involved the regulation of microcystin synthesis and cellular polyphosphate contributed to microcystin production of Microcystis responding to arsenate, since there was a positive linear correlation of the cellular microcystin quota with the exposure concentration of arsenate when the cells were not preconditioned to phosphate starvation. It is presumed that arsenate could help to actively export microcystins from living Microcystis cells when preconditioned to phosphate starvation and incubated with the medium containing 1 mu M phosphate. This study firstly provided evidence that microcystin content and/or release of Microcystis might be impacted by arsenate if it exists in harmful algal blooms. (C) 2008 Wiley Periodicals, Inc. Environ Toxicol 24:97 94, 2009.

Non-microcystin producing Microcystis wesenbergii (Komarek) Komarek (Cyanobacteria) representing a main waterbloom-forming species in Chinese waters

It is well known that several morphospecies of Microcystis, such as Microcystis aeruginosa (Kutzing) Lemmermann and Microcystis viridis (A. Brown) Lemmermann can produce hepatotoxic microcystins. However, previous studies gave contradictory conclusions about microcystin production of Microcystis wesenbergii (Komarek) Komarek. In the present study, ten Microcystis morphospecies were identified in waterblooms of seven Chinese waterbodies, and Microcystis wesenbergii was shown as the dominant species in these waters. More than 250 single colonies of M. wesenbergii were chosen, under morphological identification, to examine whether M. wesenbergii produce hepatotoxic microcystin by using multiplex PCR for molecular detection of a region (mcyA) of microcystin synthesis genes, and chemical analyses of microcystin content by ELISA and HPLC for 21 isolated strains of M. wesenbergii from these waters were also performed. Both molecular and chemical methods demonstrated that M. wesenbergii from Chinese waters did not produce microcystin. (C) 2007 Elsevier Ltd. All rights reserved.

Accumulation of hepatotoxic microcystins in freshwater mussels, aquatic insect larvae and oligochaetes in a large, shallow eutrophic lake (Lake Chaohu) of subtropical China

This paper studied the seasonal changes of two common microcystins (MCs), MC-RR and -LR, in the commercially important mussel Corbicula fluminea in Lake Chaohu, where there occurred dense cyanobacteria. Occasional measurements were also made for MC in the mussel Arconaia lanceolat, the oligochaete Limnodilus hoffineisteri and the insect larva Chironomus sp. Mean MC of C. fluminea was much higher in hepatopancreas than in intestine and foot. Our study is the first to report accumulation of MCs in oligochaetes and aquatic insect larvae. The hi-h contents of MCs in the insect larvae suggest a great possibility for the transfer of MCs to benthos-feeding omnivores like common carp. According to the provisional standard by the WHO, 28.6% of the collected C. fluminea were harmful for human consumption, assuming a daily consumption of 300 by a person. It is recommended that edible mussels should not be collected for human consumption during toxic cyanobacterial blooms in Lake Chaohu.

Detection of the hepatotoxic microcystins in 36 kinds of cyanobacteria Spirulina food products in China

Gel filtration chromatography, ultra-filtration, and solid-phase extraction silica gel clean-up were evaluated for their ability to remove microcystins selectively from extracts of cyanobacteria Spirulina samples after using the reversed-phase octadecylsilyl ODS cartridge for subsequent analysis by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). The reversed-phase ODS cartridge/silica gel combination were effective and the optimal wash and elution conditions were: H2O (wash), 20% methanol in water (wash), and 90% methanol in water (elution) for the reversed-phase ODS cartridge, followed by 80% methanol in water elution in the silica gel cartridge. The presence of microcystins in 36 kinds of cyanobacteria Spirulina health food samples obtained from various retail outlets in China were detected by LC-MS/MS, and 34 samples (94%) contained microcystins ranging from 2 to 163 ng g(-1) (mean=1427 ng g(-1)), which were significantly lower than microcystins present in blue green alga products previously reported. MC-RR-which contains two molecules of arginine (R)-(in 94.4% samples) was the predominant microcystin, followed by MC-LR-where L is leucine-(30.6%) and MC-YR-where Y is tyrose-(27.8%). The possible potential health risks from chronic exposure to microcystins from contaminated cyanobacteria Spirulina health food should not be ignored, even if the toxin concentrations were low. The method presented herein is proposed to detect microcystins present in commercial cyanobacteria Spirulina samples.

Physiological comparison between colonial and unicellular forms of Microcystis aeruginosa Kutz. (Cyanobacteria)

In order to gain insight into the bloom sustainment of colonial Microcystis aeruginosa Katz., physiological characterizations were undertaken in this study. Compared with unicellular Microcystis, colonial Microcystis phenotypes exhibited a higher maximum photosynthetic rate (Pm), a higher maximum electron transfer rate (ETRmax), higher phycocyanin content, and a higher affinity for inorganic carbon (K-0.5 DIC <= 8.4 +/- 0.7 mu M) during the growth period monitored in this study. This suggests that photosynthetic efficiency is a dominant physiological adaptation found in colonial Microcystis, thus promoting bloom sustainment. In addition, the high content of soluble and total carbohydrates in colonial Microcystis suggests that this phenotype may possess a higher ability to tolerate enhanced stress conditions when compared to unicellular (noncolonial) phenotypes. Therefore, high photosynthetic activities and high tolerance abilities may explain the bloom sustainment of colonial Microcystis in eutrophic lakes.

Applicability of the MTT assay for measuring viability of cyanobacteria and algae, specifically for Microcystis aeruginosa (Chroococcales, Cyanobacteria)

Several assay methods were screened for viability assessment in cyanobacteria using Microcystis aeruginosa FACHB 905. Compared with fluorescent diacetate (FDA), Evan's Blue and autofluorescence, the 3-[4,5-dimethylthiazol-2-yl]2,5-diphenyl tetrazolium bromide (MTT) assay, which was based on the ability of viable cells to reduce MTT to formazan, was found to be reliable and was selected for further study. MTT concentration, incubation time and temperature were optimized for M. aeruginosa. Improvements to the sensitivity and reproducibility of the MTT assay included performing it in the dark to reduce the effects of formazan light sensitivity when extracted in DMSO. Another improvement involved collecting viability data by cell by counting rather than colourimetrically, which was concluded from the fact that oxidoreductase activity, responsible for MTT reduction, would elevate or decrease under stress conditions. Half-life of oxidoreductase in dead cell was calculated to be 3 h. The MTT assay was also found to be applicable to other cyanobacteria and diatoms, including field samples, but not for algae belonging to Chlorophyta, Euglenophyta, Pyrrophyta or Chrysophyta. Based on the above results, we proposed an optimized procedure for the MTT method on Microcystis strains. The use of this assay may be of importance to better understand the dynamics of bloom and the fate of Microcystis under natural or disturbed conditions.