Technologies and Methodologies for the Detection of Harmful Algae and Their Toxins, St Petersburg, Florida, Oct 22-24 2008 : workshop proceedings

The Alliance for Coastal Technologies (ACT) Workshop "Technologies and Methodologies for the Detection of Harmful Algae and their Toxins" convened in St. Petersburg, Florida, October 22- 24, 2008 and was co-sponsored by ACT (http://act-us.info); the Cooperative Institute for Coastal and Estuarine Environmental Technology (CICEET, http://ciceet.unh.edu); and the Florida Fish and Wildlife Conservation Commission (FWC, http://www.myfwc.com). Participants from various sectors, including researchers, coastal decision makers, and technology vendors, collaborated to exchange information and build consensus. They focused on the status of currently available detection technologies and methodologies for harmful algae (HA) and their toxins, provided direction for developing operational use of existing technology, and addressed requirements for future technology developments in this area. Harmful algal blooms (HABs) in marine and freshwater systems are increasingly common worldwide and are known to cause extensive ecological, economic, and human health problems. In US waters, HABs are encountered in a growing number of locations and are also increasing in duration and severity. This expansion in HABs has led to elevated incidences of poisonous seafood, toxin-contaminated drinking water, mortality of fish and other animals dependent upon aquatic resources (including protected species), public health and economic impacts in coastal and lakeside communities, losses to aquaculture enterprises, and long-term aquatic ecosystem changes. This meeting represented the fourth ACT sponsored workshop that has addressed technology developments for improved monitoring of water-born pathogens and HA species in some form. A primary motivation was to assess the need and community support for an ACT-led Performance Demonstration of Harmful Algae Detection Technologies and Methodologies in order to facilitate their integration into regional ocean observing systems operations. The workshop focused on the identification of region-specific monitoring needs and available technologies and methodologies for detection/quantification of harmful algal species and their toxins along the US marine and freshwater coasts. To address this critical environmental issue, several technologies and methodologies have been, or are being, developed to detect and quantify various harmful algae and their associated toxins in coastal marine and freshwater environments. There are many challenges to nationwide adoption of HAB detection as part of a core monitoring infrastructure: the geographic uniqueness of primary algal species of concern around the country, the variety of HAB impacts, and the need for a clear vision of the operational requirements for monitoring the various species. Nonetheless, it was a consensus of the workshop participants that ACT should support the development of HA detection technology performance demonstrations but that these would need to be tuned regionally to algal species and toxins of concern in order to promote the adoption of state of the art technologies into HAR monitoring networks. [PDF contains 36 pages]

Concurrent exposure of bottlenose dolphins (Tursiops truncatus) to multiple algal toxins in Sarasota Bay, Florida, USA

Sentinel species such as bottlenose dolphins (Tursiops truncatus) can be impacted by large-scale mortality events due to exposure to marine algal toxins. In the Sarasota Bay region (Gulf of Mexico, Florida, USA), the bottlenose dolphin population is frequently exposed to harmful algal blooms (HABs) of Karenia brevis and the neurotoxic brevetoxins (PbTx; BTX) produced by this dinoflagellate. Live dolphins sampled during capture-release health assessments performed in this region tested positive for two HAB toxins; brevetoxin and domoic acid (DA). Over a ten-year study period (2000–2009) we have determined that bottlenose dolphins are exposed to brevetoxin and/or DA on a nearly annual basis (i.e., DA: 2004, 2005, 2006, 2008, 2009; brevetoxin: 2000, 2004, 2005, 2008, 2009) with 36% of all animals testing positive for brevetoxin (n = 118) and 53% positive for DA (n = 83) with several individuals (14%) testing positive for both neurotoxins in at least one tissue/fluid. To date there have been no previously published reports of DA in southwestern Florida marine mammals, however the May 2008 health assessment coincided with a Pseudo-nitzschia pseudodelicatissima bloom that was the likely source of DA observed in seawater and live dolphin samples. Concurrently, both DA and brevetoxin were observed in common prey fish. Although no Pseudo-nitzschia bloom was identified the following year, DA was identified in seawater, fish, sediment, snails, and dolphins. DA concentrations in feces were positively correlated with hematologic parameters including an increase in total white blood cell (p = 0.001) and eosinophil (p<0.001) counts. Our findings demonstrate that dolphins within Sarasota Bay are commonly exposed to two algal toxins, and provide the impetus to further explore the potential long-term impacts on bottlenose dolphin health.

Characterizing a cyanobacterial bloom in western Lake Erie using satellite imagery and meteorological data

The distribution and intensity of a bloom of the toxic cyanobacterium, Microcystis aeruginosa, in western Lake Erie was characterized using a combination of satellite ocean-color imagery, field data, and meteorological observations. The bloom was first identified by satellite on 14 August 2008 and persisted for more than 2 months. The distribution and intensity of the bloom was estimated using a satellite algorithm that is sensitive to near-surface concentrations of M. aeruginosa. Increases in both area and intensity were most pronounced for wind stress less than 0.05 Pa. Area increased while intensity did not change for wind stresses of 0.05–0.1 Pa, and both decreased for wind stress greater than 0.1 Pa. The recovery in intensity at the surface after strong wind events indicated that high wind stress mixed the bloom through the water column and that it returned to the surface once mixing stopped. This interaction is consistent with the understanding of the buoyancy of these blooms. Cloud cover (reduced light) may have a weak influence on intensity during calm conditions. While water temperature remained greater than 15°C, the bloom intensified if there were calm conditions. For water temperature less than 15°C, the bloom subsided under similar conditions. As a result, wind stress needs to be considered when interpreting satellite imagery of these blooms.

Blood cells as targets of snake toxins

Snake venoms are mixtures of enzymes and peptides which exert toxicological effects by targeting their substrates or receptors upon envenomation. Snake venom proteins widely affect vascular system including circulating blood cells, coagulation factors, an

Some marine toxins

The article presents some toxins of marine origin. These are tetrodotoxin from pufferfish, saxitoxins from red microalgae that mussels and shellfish ingest, and palitoxin the most toxic marine product isolated from zoonthurian Palythoa toxica.

Effects of toxins induced by phytoplankton bloom on physiological function of indicator animals in Anzali Lagoon

The present overview summarizes data from one year's study during the period of 1379 to 1380 in the regions by "Anzali" lagoon called "Abkenar" and "Hendkhale". Specimens from this lagoon obtained weekly during mordad and shahrivar mounths (July 21 to september 21). The study included 67 types of 5 phytoplanktonal phylum. In "Abkenar" region Cyanobacters with maximum of 97% and minimum of 64.5% of total combination of phytoplanktones made the dominating combination during the period of study , while in "hendkhale" chyrsophyta with maximum of 89% and minimum of 38.7% of total phytoplanktonal was the dominating figure at the same period of time. Researches on ecological parameteres showed that the avarage dissolved oxygen in -Abkenar" and "hendlchale" regions was 10.7 and 8.0 mg/lit respectivly, also total rate of Phosphat in these regions was 0.085 and 0.15 mg/lit respectivly. This study showed that the rates of Nitrat and Amonium in "Abkenar" region was 0.043 and 0.79 mg / lit while for the same substances in "Ilendkhale" measured 0.08 and 0.7 mg I lit respective. Also the avarage rate of chlorophyll a in these two areas measured 58,38 and 40.45 j.un /ht respectivly. Depending on results of correlation cofficient in "Abkenar" region we had Cyanobacters , water and air temperature , Chlorophyll a and total amount of Phosphat as a poitive correlation with transparency while Amonium and Nitrat showed , a negative correlation , EC and finally dissolved oxygen showed a very low rate of correlation coffiocient. To perform this research 5 genus of Cyanobacteres horn "Anzali" lagoon have been isolated and cultured in a laboratorial conditions Later by using Mouse Bioassay method one of these genus identified as a toxic algae. Levels of LD50 with intra peritoneal injection of toxin on mouse in 24 hours was 660 mg/kg and Levels of LC50 by using Bioassay method on Artemia and Daphnia has been shown 618 and 1000 mg kg respectively. Also the physiological effects of were investigated. algae on two types of Cypronides family called Cyprinus carpi° and Hypophrthabnichthys Resultes of blood analyses of Cypronides who were feeded by toxic algae showed a significant decline (P < 0.05) in white and red blood cells and their hematocrites. Levels of LDH , SGOT and SGPT in their blood serum had a significant increase in porportion to control group (P < 0.05) but there was no evidance a differences in Total Protein levels. Pathological studies show damage and destruction of hepato pancreas and kidney of these fishes. Signs and symptoms of intoxication caused by Cyanobacter called Planktohrix agardlltii in mouse and fish show heptotoxic character. This toxin belongs to cyclicpeptides of microcystines group.

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.

Influence of dew on biomass and photosystem II activity of cyanobacterial crusts in the Hopq Desert, northwest China

Dew is an important water source for desert organisms in semiarid and arid regions. Both field and laboratory experiments were conducted to investigate the possible roles of dew in growth of biomass and photosynthetic activity within cyanobacterial crust. The cyanobacteria, Microcoleus vaginatus Gom. and Scytonema javanicum (Kutz.) Born et Flah., were begun with stock cultures and sequential mass cultivations, and then the field experiment was performed by inoculating the inocula onto shifting sand for forming cyanobacterial crust during late summer and autumn of 2007 in Hopq Desert, northwest China. Measurements of dew amount and Chlorophyll a content were carried out in order to evaluate the changes in crust biomass following dew. Also, we determined the activity of photosystem II(PSII) within the crust in the laboratory by simulating the desiccation/rehydration process due to dew. Results showed that the average daily dew amount as measured by the cloth-plate method (CPM) was 0.154 mm during fifty-three days and that the crust biomass fluctuated from initial inoculation of 4.3 mu g Chlorophyll a cm(-2) sand to 5.8-7.3 mu g Chlorophyll a cm(-2) crust when dew acted as the sole water source, and reached a peak value of approximately 8.2 mu g Chlorophyll a cm(-2) crust owing to rainfalls. It indicated that there was a highly significant correlation between dew amounts and crust moistures (r = 0.897 or r = 0.882, all P < 0.0001), but not a significant correlation between dew and the biomass (r = 0.246 or r = 0.257, all P > 0.05), and thus concluded that dew might only play a relatively limited role in regulating the crust biomass. Correspondingly, we found that rains significantly facilitated biomass increase of the cyanobacterial crust. Results from the simulative experiment upon rehydration showed that approximately 80% of PSII activity could be achieved within about 50 min after rehydration in the dark and at 5 degrees C, and only about 20% of the activity was light-temperature dependent. This might mean that dew was crucial for cyanobacterial crust to rapidly activate photosynthetic activity during desiccation and rehydration despite low temperatures and weak light before dawn. It also showed in this study that the cyanobacterial crusts could receive and retain more dew than sand, which depended on microclimatic characteristics and soil properties of the crusts. It may be necessary for us to fully understanding the influence of dew on regulating the growth and activity of cyanobacterial crust, and to soundly evaluate the crust's potential application in fighting desertification because of the available water due to dew. (C) 2009 Published by Elsevier Ltd.

Rapid quantification of total microcystins in cyanobacterial samples by periodate-permanganate oxidation and reversed-phase liquid chromatography

Microcystins (MCs) comprise a family of more than 80 related cyclic hepatotoxic heptapeptides. Oxidation of MCs causes cleavage of the chemically unique C-20 beta-amino acid (2S, 3S, 8S, 9S)-3-amino-9-methoxy-2,6,8-trimethyl-10-phenyldeca-4,6-dienoic acid (Adda) amino to form 2-methyl-3-methoxy-4-phenylbutanoic acid (MMPB), which has been exploited to enable analysis of the entire family. In the present study, the reaction conditions (e.g. concentration of the reactants. temperature and pH) used in the production of MMPB by oxidation of cyanobacterial samples with permanganate-periodate were optimized through a series of well-controlled batch experiments. The oxidation product (MMPB) was then directly analyzed by high-performance liquid chromatography with diode array detection. The results of this study provided insight into the influence of reaction conditions on the yield of MMPB. Specifically, the optimal conditions, including a high dose of permanganate (>= 50 mM) in saturated periodate solution at ambient temperature under alkaline conditions (pH similar to 9) over 1-4 h were proposed, as indicated by a MMPB yield of greater than 85%. The technique developed here was applied to determine the total concentration of MCs in cyanobacterial bloom samples, and indicated that the MMPB technique was a highly sensitive and accurate method of quantifying total MCs. Additionally, these results will aid in development of a highly effective analytical method for detection of MMPB as an oxidation product for evaluation of total MCs in a wide range of environmental sample matrices, including natural waters, soils (sediments) and animal tissues. (C) 2009 Elsevier B.V. All rights reserved.

Effects of cyanobacterial toxin microcystin-LR on the transcription levels of immune-related genes in grass carp Ctenopharyngodon idella

Recent studies in mammals have revealed that the cyanobacterial toxin MC-LR suppresses immune functions. Nevertheless, immunotoxic effects of microcystins have been little studied in fish. In this paper, we present the profiles of the immune modulation of MC-LR in grass carp, and quantitative real-time PCR methodology was developed for the measurement of relative transcription changes of six immune-related genes in the spleen and head kidney of the grass carp Ctenopharyngodon idella, which were intraperitoneally injected with 50 mu g MC-LR center dot kg(-1) body weight in a three-week period. This study was focused exclusively on gene transcription level changes at different time points after MC-LR exposure, so, only one dose was given. The investigated genes were interleukin-1 beta (IL-1 beta), tumor necrosis factor-alpha (TNF-alpha), type I interferon (Type I IFN), peptidoglycan recognition protein-L (PGRP-L), immunoglobulin M (IgM) and major histocompatibility complex class I (MHC-I) genes. The results demonstrated that the transcription levels of the TNF-alpha, type I IFN, and PGRP-L genes in the spleen and head kidney were significantly low at all time points, and those of IL-1 beta were significantly low in the head kidney at different time points. In addition, IgM and MHC-I transcription levels were only significantly low in the spleen and head kidney at 21 d postinjection. The changes in the transcription levels of immune-related genes induced by MC-LR confirmed its effect on inhibiting immune function at the transcription level.

Plankton Community Succession in Artificial Systems Subjected to Cyanobacterial Blooms Removal using Chitosan-Modified Soils

Using artificial systems to simulate natural lake environments with cyanobacterial blooms, we investigated plankton community succession by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) fingerprinting and morphological method. With this approach, we explored potential ecological effects of a newly developed cyanobacterial blooms removal method using chitosan-modified soils. Results of PCR-DGGE and morphological identification showed that plankton communities in the four test systems were nearly identical at the beginning of the experiment. After applying the newly developed and standard removal methods, there was a shift in community composition, but neither chemical conditions nor plankton succession were significantly affected by the cyanobacteria removal process. The planted Vallisneria natans successfully recovered after cyanobacteria removal, whereas that in the box without removal process did not. Additionally, canonical correspondence analysis indicated that other than for zooplankton abundance, total phosphorus was the most important environmental predictor of planktonic composition. The present study and others suggest that dealing with cyanobacteria removal using chitosan-modified soils can play an important role in controlling cyanobacterial blooms in eutrophicated freshwater systems.

Feasibility of cyanobacterial inoculation for biological soil crusts formation in desert area

Practical testing of the feasibility of cyanobacterial inoculation to speed up the recovery of biological soil crusts in the field was conducted in this experiment. Results showed that cyanobacterial and algal cover climbed up to 48.5% and a total of 14 cyanobacterial and algal species were identified at the termination of inoculation experiment; biological crusts' thickness, compressive and chlorophyll a content increased with inoculation time among 3 years; moss species appeared in the second year; cyanobacterial inoculation increased organic carbon and total nitrogen of the soil; total salt, calcium carbonate and electrical conductivity in the soil also increased after inoculation. Diverse vascular plant communities composed of 10 and 9 species are established by cyanobacterial inoculation on the windward and leeward surface of the dunes, respectively, after 3 years. The Simpson index for the above two communities are 0.842 and 0.852, while the Shannon-Weiner index are 2.097 and 2.053, respectively. In conclusion, we suggest that cyanobacterial inoculation would be a suitable and effective technique to recover biological soil crusts, and may further restore the ecological system. (C) 2008 Published by Elsevier Ltd.

Plasma biochemical responses of the planktivorous filter-feeding silver carp (Hypophthalmichthys molitrix) and bighead carp (Aristichthys nobilis) to prolonged toxic cyanobacterial blooms in natural waters

The planktivorous filter-feeding silver carp (Hypophthalmichthys molitrix) and bighead carp (Aristichthys nobilis) are the attractive candidates for bio-control of plankton communities to eliminate odorous populations of cyanobacteria. However, few studies focused on the health of such fishes in natural water body with vigorous toxic blooms. Blood parameters are useful and sensitive for diagnosis of diseases and monitoring of the physiological status of fish exposed to toxicants. To evaluate the impact of toxic cyanobacterial blooms on the planktivorous fish, 12 serum chemistry variables were investigated in silver carp and bighead carp for 9 months, in a large net cage in Meiliang Bay, a hypereutrophic region of Lake Taihu. The results confirmed adverse effects of cyanobacterial blooms on two phytoplanktivorous fish, which mainly characterized with potential toxicogenomic effects and metabolism disorders in liver, and kidney dysfunction. In addition, cholestasis was intensively implied by distinct elevation of all four related biomarkers (ALP, GGT, DBIL, TBIL) in bighead carp. The combination of LDH, AST activities and DBIL, URIC contents for silver carp, and the combination of ALT. ALP activities and TBIL, DBIL. URIC concentrations for bighead carps were found to most strongly indicate toxic effects from cyanobacterial blooms in such fishes by a multivariate discriminant analysis. (C) 2009 Elsevier B.V. All rights reserved.

Interactions between a cyanobacterial bloom (Microcystis) and the submerged aquatic plant Ceratophyllum oryzetorum Kom.

In aquatic ecosystems, macrophytes and phytoplankton are main primary producers, in which macrophyte plays an important role in maintaining clear water state, while phytoplankton often dominates in turbid waterbodies. In the present study, the growth and photosynthetic activity of the submerged aquatic plant Ceratophyllum oryzetorum Kom. in different cell densities of cyanobacterial bloom are studied. The results show that the plant length and fresh mass of C. oryzetorum are promoted by low cyanobacterial cell densities. Medium and high cyanobacterial cell densities, on the contrary, act as inhibitory. Furthermore, the photosynthetic activity of C. oryzetorum is strongly inhibited by high cyanobacterial cell densities. To a certain extent, the growth of cyanobacteria is inhibited by C. oryzetorum, but no significant effect is found in this study.