New Zealand Guidelines for cyanobacteria in recreational fresh waters: Interim Guidelines

This document is divided into four main sections, plus 14 appendices. Section 1. Introduction provides an overview of the purpose and status of the document as well as advice on who should use it. Section 2. Framework provides a background to the overall guidelines approach, recommendations on agency roles and responsibilities, and information on the condition of use of this document. Section 3. Guidelines describes the recommended three-tier monitoring and action sequence for planktonic and benthic cyanobacteria. Section 4. Sampling provides advice on sampling planktonic and benthic cyanobacteria. The appendices give further background information and include templates for data collection and reporting, including: • background information on known cyanotoxins and their distribution in New Zealand • information on the derivation of guideline values • photographs of typical bloom events • a list of biovolumes for common New Zealand cyanobacteria • templates for field assessments • suggested media releases and warning sign templates. A glossary provides definitions for abbreviations and terms used in these guidelines

Recreational exposure during algal bloom in carrasco beach, uruguay: A liver failure case report

In January 2015, a 20-month-old child and her family took part in recreational activities at Carrasco and Malvín beaches (Montevideo, Uruguay). An intense harmful algae bloom (HAB) was developing along the coast at that time. A few hours after the last recreational exposure episode, the family suffered gastrointestinal symptoms which were self-limited except in the child’s case, who was admitted to hospital in Uruguay with diarrhea, vomiting, fatigue, and jaundice. The patient had increased serum levels of liver enzymes and bilirubin and five days later presented acute liver failure. She was referred to the Italian Hospital in Buenos Aires, being admitted with grade II-III encephalopathy and hepatomegaly and requiring mechanical respiratory assistance. Serology tests for hepatitis A, B, and C, Epstein-Barr virus, and cytomegalovirus were negative. Laboratory features showed anemia, coagulopathy, and increased serum levels of ammonium, alanine aminotransferase (ALT), aspartate aminotransferase (AST), and bilirubin. Autoimmune Hepatitis Type-II (AH-II) was the initial diagnosis based on a liver kidney microsomal type 1 antibodies (LKM-1) positive result, and twenty days later a liver transplant was performed. The liver histopathology had indicated hemorrhagic necrosis in zone 3, and cholestasis and nodular regeneration, which were not characteristic of AH-II. LC/ESI-HRMS (liquid chromatography electrospray ionization high-resolution mass spectrometry) analysis of MCs in the explanted liver revealed the presence of Microsytin-LR (MC-LR) (2.4 ng·gr-1 tissue) and [D-Leu1]MC-LR (75.4 ng·gr-1 tissue), which constitute a toxicological nexus and indicate a preponderant role of microcystins in the development of fulminant hepatitis.Fil: Vidal, Flavia. Hospital Italiano; ArgentinaFil: Sedan, Daniela Yazmine. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Departamento de Química. Centro de Investigaciones del Medio Ambiente; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: D'Agostino, Daniel. Hospital Italiano; ArgentinaFil: Cavalieri, María Lorena. Hospital Italiano; ArgentinaFil: Mullen, Eduardo. Hospital Italiano; ArgentinaFil: Parot Varela, María Macarena. Hospital Italiano; ArgentinaFil: Flores, Cintia. Instituto de Diagnóstico Ambiental y Estudios del Agua; EspañaFil: Caixach, Josep. Instituto de Diagnóstico Ambiental y Estudios del Agua; EspañaFil: Andrinolo, Dario. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Departamento de Química. Centro de Investigaciones del Medio Ambiente; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

Microcystins in components of twelve New Hampshire lakes of varied trophic status

Cyanobacteria toxins, called microcystins (MCs), were found in components of twelve, stratified New Hampshire (USA) lakes of varied trophic status during the summer of 1998. A sensitive ELISA method detected MC levels in whole lakewater, grazable phytoplankton fractions (\u3c30 mm), net phytoplankton (\u3e 375 µm), and isolated copepod and cladoceran (Daphnia sp.) samples. Whole lakewater (WLW) and net phytoplankton MC concentrations ranged between 9 and 165 ng MC L-1 and 0.2 and 2031 mg MC g-1-dry wt, respectively. Lakewater MC concentrations correlated with total epilimnetic phosphorus and total epilimnetic chlorophyll a concentrations and inversely with Secchi disk depth. The filter-feeding cladoceran (daphnid) and omnivorous copepod components of the zooplankton were separated and assayed independently for MCs. The cladoceran component accumulated between 7 and 2800 µg MC g-1-dry wt. in 10 of the lakes. The copepod component accumulated similar levels (4 and 2400 µg MC g-1 dry-wt.) in all lakes. Toxin accumulation by zooplankton directly correlated with lakewater and net phytoplankton MC concentrations. The highest levels were found in Silver Lake, a productive lake where Microcystis aeruginosa blooms frequently occur. It is particularly noteworthy that MC levels were also detected by ELISA methods in Russell Pond, a pristine, deep mountain lake of low productivity. The results emphasize the importance of including oligotrophic lakes and water supplies in monitoring programs for MCs to ensure the safety of animals and humans utilizing them for drinking and recreation

New Method for Simultaneous Determination of Microcystins and Cylindrospermopsin in Vegetable Matrices by SPE-UPLC-MS/MS

Cyanotoxins are a large group of noxious metabolites with different chemical structure and mechanisms of action, with a worldwide distribution, producing effects in animals, humans, and crop plants. When cyanotoxin-contaminated waters are used for the irrigation of edible vegetables, humans can be in contact with these toxins through the food chain. In this work, a method for the simultaneous detection of Microcystin-LR (MC-LR), Microcystin-RR (MC-RR), Microcystin-YR (MC-YR), and Cylindrospermopsin (CYN) in lettuce has been optimized and validated, using a dual solid phase extraction (SPE) system for toxin extraction and ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) for analysis. Results showed linear ranges (5–50 ng g−1 f.w.), low values for limit of detection (LOD) (0.06–0.42 ng g−1 f.w.), and limit of quantification (LOQ) (0.16–0.91 ng g−1 f.w.), acceptable recoveries (41–93%), and %RSDIP values for the four toxins. The method proved to be robust for the three variables tested. Finally, it was successfully applied to detect these cyanotoxins in edible vegetables exposed to cyanobacterial extracts under laboratory conditions, and it could be useful for monitoring these toxins in edible vegetables for better exposure estimation in terms of risk assessment.España MINECO AGL2015-64558-

Analysis of the Use of Cylindrospermopsin and/or Microcystin-Contaminated Water in the Growth, Mineral Content, and Contamination of Spinacia oleracea and Lactuca sativa

Cyanobacteria and cyanotoxins constitute a serious environmental and human health problem. Moreover, concerns are raised with the use of contaminated water in agriculture and vegetable production as this can lead to food contamination and human exposure to toxins as well as impairment in crop development and productivity. The objective of this work was to assess the susceptibility of two green vegetables, spinach and lettuce, to the cyanotoxins microcystin (MC) and cylindrospermopsin (CYN), individually and in mixture. The study consisted of growing both vegetables in hydroponics, under controlled conditions, for 21 days in nutrient medium doped with MC or CYN at 10 μg/L and 50 μg/L, or CYN/MC mixture at 5 + 5 μg/L and 25 + 25 μg/L. Extracts from M. aeruginosa and C. ovalisporum were used as sources of toxins. The study revealed growth inhibition of the aerial part (Leaves) in both species when treated with 50µg/L of MC, CYN and CYN/MC mixture. MC showed to be more harmful to plant growth than CYN. Moreover spinach leaves growth was inhibited by both 5 + 5 and 25 + 25 µg/L CYN/MC mixtures, whereas lettuce leaves growth was inhibited only by 25 + 25 µg/L CYN/MC mixture. Overall, growth data evidence increased sensitivity of spinach to cyanotoxins in comparison to lettuce. On the other hand, plants exposed to CYN/MC mixture showed differential accumulation of CYN and MC. In addition, CYN, but not MC, was translocated from the roots to the leaves. CYN and MC affected the levels of minerals particularly in plant roots. The elements most affected were Ca, K and Mg. However, in leaves K was the mineral that was affected by exposure to cyanotoxinsEspaña, MINECO/FEDER Project AGL2015-64558-

Further characterization of glycine-containing microcystins from the McMurdo Dry Valleys of Antarctica

Microcystins are hepatotoxic cyclic peptides produced by several cyanobacterial genera worldwide. In 2008, our research group identified eight new glycine-containing microcystin congeners in two hydro-terrestrial mat samples from the McMurdo Dry Valleys of Eastern Antarctica. During the present study, high-resolution mass spectrometry, amino acid analysis and micro-scale thiol derivatization were used to further elucidate their structures. The Antarctic microcystin congeners contained the rare substitution of the position-1 D-alanine for glycine, as well as the acetyl desmethyl modification of the position-5 Adda moiety (3S-amino-9S-methoxy-2S,6,8S-trimethyl-10-phenyldeca-4E,6E-dienoic acid). Amino acid analysis was used to determine the stereochemistry of several of the amino acids and conclusively demonstrated the presence of glycine in the microcystins. A recently developed thiol derivatization technique showed that each microcystin contained dehydrobutyrine in position-7 instead of the commonly observed N-methyl dehydroalanine

Increasing Microcystis cell density enhances microcystin synthesis: a mesocosm study

An experimental protocol using mesocosms was established to study the effect of Microcystis sp. cell abundance on microcystin production. The mesocosms (55 L) were set up in a shallow eutrophic lake and received either no (control), low (to simulate a moderate surface accumulation), or high (to simulate a dense surface scum) concentrations of Microcystis sp. cells collected from the lake water adjacent to the mesocosms. In the low- and high-cell addition mesocosms (2 replicates of each), the initial addition of Microcystis sp. cells doubled the starting cell abundance from 500 000 to 1 000 000 cells mL⁻¹, but there was no detectable effect on microcystin quotas. Two further cell additions were made to the high-cell addition mesocosms after 60 and 120 min, increasing densities to 2 900 000 and 7 000 000 cells mL-1, respectively. Both additions resulted in marked increases in microcystin quotas from 0.1 pg cell-1 to 0.60 and 1.38 pg cell⁻¹, respectively, over the 240 min period. Extracellular microcystins accounted for <12% of the total microcystin load throughout the whole experiment. The results of this study indicate a relationship between Microcystis cell abundance and/or mutually correlated environmental parameters and microcystin synthesis

Occurrence and toxicity of Microcystis aeruginosa (Cyanobacteria) in the Paraná River, downstream of the Yacyretá dam (Argentina)

Cyanobacteria constitute the main toxin producers in inland water ecosystems and have extensive global distribution. The presence of hepatotoxins in aquatic environments is hazardous to human and animal health; even though the presence and identification of hepatotoxic microcystins in rivers and reservoirs of the world have been confirmed by several studies in the last few years. Herein, we studied the abundance and toxicity of Microcystis aeruginosa in the Argentine section of the Paraná River at the beginning of the Middle Paraná (Corrientes Hydrometer), near Corrientes city (27º28´ S - 58º51´ W) and approximately 220 km downstream of the Yacyretá dam (High Paraná). The Paraná River basin, with a drainage area of 3.1 x 106 km2 and 3 965 km in length, is the second largest catchment of South America, after that of the Amazon. The Paraná River is the main source of drinking water supply for the Northeastern Argentine region. Phytoplankton samples were collected and environmental variables were measured in a monthly basis (exceptionally fortnightly), from March 2004 to June 2008. Fifty-eight samples were analyzed for phytoplankton density and biomass. Five samples were used for toxicity testing; the latter were obtained during the cyanobacteria blooms from 2005 to 2008. Phytoplankton counts were performed with an inverted microscope, and biomass was expressed as biovolume. Bioassays with mice and high-performance liquid chromatography (HPLC) analysis were performed to evaluate the presence of cyanotoxins. Phytoplankton mainly consisted of Cryptophyta, Chlorophyta and Bacillariophyta. Microcystis aeruginosa was identified during the warmer months each year (November to March). Density varied between 189 and 25 027 cells/mL (1-10 colonies/mL) and biomass from 0.34 to 44 mm3/L. Taking into account the number of cells, the highest abundance occurred in April 2004 (25 027 cells/mL), coinciding with the largest biovolume (44 mm3/L). All mice subjected to intraperitoneal injections with samples obtained during bloom episodes showed positive results for the presence of hepatotoxins. Three microcystins variants: LR, RR and [D-Leu1] Mcyst-LR were detected by analysis with semi-preparative high-performance liquid chromatography with diode array detector system (HPLC-PDA). This constitutes the first report of microcystins recorded during M. aeruginosa blooms in the Argentine stretch of the Paraná River at the beginning of the Middle Paraná (Corrientes Hydrometer), approximately 220 km downstream of the Yacyretá dam (High Paraná).Fil: Forastier, Marina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Centro de Ecología Aplicada del Litoral. Universidad Nacional del Nordeste. Centro de Ecología Aplicada del Litoral; ArgentinaFil: Zalocar, Yolanda. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Centro de Ecología Aplicada del Litoral. Universidad Nacional del Nordeste. Centro de Ecología Aplicada del Litoral; ArgentinaFil: Andrinolo, Dario. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Exactas; ArgentinaFil: Domitrovic, Hugo Alberto. Universidad Nacional del Nordeste. Facultad de Ciencias Veterinarias; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin