Cyanobacteria and cyanotoxins studies in some parts of Russian Federation
Ekaterina Chernova , Zoya Zhakovskaya
Saint-Petersburg Scientific Research Centre for Ecological Safety Russian Academy of Sciences (SRCES RAS) 19-21 February 2015, Seville Saint-Petersburg Scientific Research Centre for Ecological Safety Russian Academy of Sciences
Our department - Eco-Chemical studies
Head of the department Zoya Zhakovskaya, phD zazhak@hotmail,com
Algological Studies EkaterinaVoyakina, phD [email protected] Instrumental analysis (LCMS) – cyanotoxins determination Iana Russkikh, phD, [email protected] Total neurotoxicity (biosensors) Ekaterina Chernova Anna Pilip [email protected] [email protected] Saint-Petersburg SRCES RAS
Area– 11,0 km2 Area– 0,98 km2
Sestroretskij Low Suzdal Razliv Lake Lake
Monitoring of HAB of two eutrophic Saint Petersburg lakes with high recreation load since 2008. Saint-Petersburg SRCES RAS Toxin-producing species in water bodies of Saint-Petersburg
M. viridis Microcystis spp. M. aeruginosa
M. wesenbergii Aphanizomenon flos-aquae Planktothrix agardhii Saint-Petersburg SRECES RAS Sestroretskij Razliv Lake a large artificial eutrophic reservoir primarily used for recreational purposes.
Surface area – 11,0 km2, average depth – 1,6 m.
Nutrients gradients: ammonia nitrogen (0,6 – 2,0 mg N L–1) phosphates (0,002 – 0,600 mg P L–1). Cyanobacterial biomass: the highest - 182,7 mg L–1 in 2013 Cyanobacteria contribution in phytoplankton varied from 30% to 88% of the total biomass during 2010–2014. HAB in Sestroretskij Razliv Lake, 2010
Dominated species: Aphanizomenon flos-aquae, Planktothrix agardhii, Microcystis aeruginosa, Detected MC variants: M.wesenbergii, M. viridis MC-LR, dm-MC-LR, MC-RR, dm-MC-RR, MC-YR Average seasonal ΣMCs 0,2-6,3 μg L-1 St-Petersburg, SRCES RAS The surface area is 0,97 km2, Lower Suzdal Lake the average depth is 3,0 m. Cyanobacteria biomass Average – 6,2 mg L–1 the highest – 26,0 mg L–1 in 2012 Cyanobacteria contribution in phytoplankton 10-70% of the total biomass (2010–2014).
Dominated species: Aphanizomenon flos-aquae, Planktothrix agardhii, Microcystis spp. Detected cyanotoxins by LCMS: average seasonal ΣMCs 0,1-0,9 μg L-1 dm-MC-RR, MC-RR and MC-YR – the main MC variants.
2014: Aphanizomenon flos-aquae periodically was monodominant species . Anatoxin-A in water was determined during May-October, 2014 St-Petersburg, SRCES RAS Determination of Total neurotoxicity using biosensors: target compound - AN-A(S) as organophosphate
Amperometric method is used for the detection of AN-A(S) by the estimation of Cholinesterase Activity after influence of potential inhibitors using the equipment “Easy Check” .
Principle of method: Certain chemical classes of compounds, such as organophosphates (OPs) and carbamates (CMs) can 'inhibiting' cholinesterase.
Necessary to run differential analysis for carbamates and organophosphates pesticides by another method (for instance, LCMS)
LOD of method 0.5-2.0 nmol of OPs Analysis Time - 30 min Sample volume – 1ml
Consumables: enzymes (ChE) and substrates (propionyl thyoholine) St-Petersburg, SRCES RAS Laboratory of biological methods for ecological safety Head of Laboratory - Nadezda Medvedeva, Dr. Sci., ([email protected]) Applied methods: Microbiological, Biochemical, Instrumental physico-chemical methods Research works: * Effect of pollutants on Growth, Development and Toxin Production by Microalgae The stress response of microalgae (genera Anabaena, Aphanizomenon, Microcystis, Nodularia, Planktothrix, Oocystis, Scenedesmus) on the impact of organic and inorganic pollutants was investigated. Result: heavy metals, 1,2,4-triazole, octyl- and nonylphenols influenced on microcystin production by toxigenic cyanobacteria of genera Microcystis, Planktothrix. * Microbiological destruction of algal toxins Active bacterial strains - destructors of microcystins MC-LR, dmMC-LR, MC- RR were isolated from natural sources. The enzymatic pathway for the degradation of these hepatotoxins are studied. The Institutions of Russian Federation running research work on cyanobacteria Scientific Research Centre for Ecological Safety Russian Academy of Sciences Apatity • KaliningradPhytoplankton Studies (blue-green algae) * * St-Petersburg * +карты озер * Syktyvkar Low Suzdal * Rybinsk Lake: MoscowBiomass * in *summerYaroslavl period:______Dominated species Detected concentration of hepatotoxins МС-LR Total MCs MC variants
* Irkutsk Laboratoty for Marine Ecology, Atlantic Branch of P.P.Shirshov Institute of Oceanology RAS Kaliningrad Study of cyanotoxins and its ecological effect Study of HABs and cyanotoxins Elena Ezhova, PhD, Head of Laboratory ([email protected]) (bloom dynamics, (2010 -2014) has been carried out at the experimental ecotoxicology) shallow lagoons of South-Eastern Baltic Eugenia Lange, researcher (phytoplankton and in the marine areas. composition & dynamics, toxic species) Methods: HAB monitoring Maria Maksimova, researcher (cell culturing, toxin’s biodegradation) (hydrochemistry / biology), Natalia Molchanova, researcher & Julia Polunina, field screening of cyanotoxins(ELISA), PhD, senior researcher (experimental ecotoxicology) phytoplankton study, experiment. LME AB IORAS collaborate with: Center of Ecological Safety RAS (St.-Petersburg): hepatotoxins and AN-a determination by LCMS “Curonian Spit” National Park (Kalinigrad region): HABs monitoring, public relation activity, “Stylab Ltd.” (Moskow): methods of cyanotoxin http://www.park-kosa.ru/cn_novosti/publikatsii/?ELEMENT_ID=59695 http://www.park-kosa.ru/cn_novosti/publikatsii/?ELEMENT_ID=63939 screening http://www.park-kosa.ru/cn_novosti/publikatsii/?ELEMENT_ID=66069 Vistula Lagoon & Curonian Lagoon shallow water bodies– av. 2,7 m, max. ca. 5 m Curonian mostly freshwater, Vistula Brackish 3-5 PSU Curonian - hyper-trophic, Vistula – eutrophic Basins of high-productive commercial fishery (bream, herring, eel, whitefish) Recreational area (national park/reserves, tourism)
Phytoplankton biomass can locally be over 1000 g m-3, average 10 g m-3.
The bloom-forming algae - potentially toxic cyanobacterial species of Aphanizomenon, Anabaena, Microcystis, Planktotrix, Woronichinia genera (in Curonian) and also Nodularia (in Vistula). MCs concentrations in waters of Curonian Lagoon, Kaliningrad (LCMS method)
Water sample, Biomass sample, Dominated μg L-1 μg g-1 (d.w.) species ΣМСs МС - LR ΣМСs МС - LR 2010 qualitative analysis, Aph. flos-aquae, Arg –containing MCs variants were detected Microcystis spp. 2011 1,0 - 194,8 2012 0,04 - 18,8 0,01 - 3,3 1,3 - 658,2 0,1 - 151,5 Aph. flos-aquae, Microcystis spp 152,6 - 290,5* 25,0 – 65,5* 2013 2,2 - 14,9 0,1 - 0,5 25,0 – 4719,0 0,3 – 77,0 Aph. flos-aquae, Microcystis spp * Pelagic zone Curonian Lagoon, Blooming of 2010 Kaliningrad, Curonian and Vistula Lagoons Fast death of piscivorous birds after consuming of dead fish Cases of fish and animals mass mortality occur yearly, up to 2-3 times per year. Fish-kills - May, Jul-Aug, Sept Piscivorous birds – Jul-Aug; Invertebrate-kills (mollusks of Planorbiddae, Lymnaeidae, Viviparidae, paralysis of Chironomus plumosus larvae) – Jul-Aug Kaliningrad, Curonian and Vistula Lagoons Every summer period high concentrations of MCs were detected. During bloom of Aphanizomenon flos-aqua in Oct. 2013 clear neurotoxic effect was proved by experiments on Daphnia. Experiment : Survival of Daphnia magna in different dilution of filtrate of natural water sample, Curonian Lagoon, 06.10.2013 Phytoplankton cells from Curonian lagoon Rybinsk, Borok, Volga Reservoirs I.D. Papanin Institute for biology of inland waters, Russian Academy of Sciences (www.ibiw.ru) • Laboratory of algology • Laboratory of microbiology Joint work with - Saint-Petersburg SRCES RAS on LCMS determination of MCs - Space Research Institute ,Russian Academy of Sciences, Moscow (satellite photoes) - P.G. Demidov Yaroslavl State University on PCR analysis Volga basin Phytoplankton monitoring of Volga Reservoirs has run since 1954. Rybinskoje Different trophic status of reservoirs: Mesotrophic and eutrophic. Chebocsarskoje 280 cyanobacterial species observed Dominated species: Aphanizomenon flos-aquae, Gorkovskoje Microcystis aeruginosa, Anabaena spp. (A. flos- aquae, A.scheremetievi, A.spiroides, A. lemmermannii ). Planktothrix agardhii has become dominant species in the oldest Ivankovskij reservoir (since the beginning of 1970s) after creating Volga reservoir system. Volga Reservoirs LCMS analysis: occurrence of MCs in all water samples. Detected up to 17 MCs variants. Main toxins variants: MC-LR, MC-RR, dmMC-RR, MC-YR Yaroslavl State University, Department of ecology and zoology (www.uniyar.ac.ru/en/) Scientific Interests: hepato- and neurotoxigenic cyanobacteria, ecological role of cyanotoxins, PCR detection of toxic cyanobacteria, ecology of shallow lakes, phytoplankton Team: The head: Babanazarova Olga, Zubishina Alla, ELISA algology, phytoplankton studies Sidelev Sergey, PCR analysis ([email protected]) ([email protected]) ([email protected]) Applied methods: conventional PCR, real-time PCR, RFLP- analysis, PCR with individual colonies of toxigenic cyanobacteria, multiplex PCR, ELISA, fluorescence microscopy Yaroslavl State University, Department of ecology and zoology Principle fields of research: Molecular genetics identification of microcystin-, cylindrospermopsin-, anatoxin-a-, PSP-producing cyanobacteria in the Russian freshwaters. Experimental study of the ecological role of cyanobacterial toxins using natural mesocosms: testing allelopathic hypothesis, signaling hypothesis and the protective role against zooplankton grazing. Detection of microcystins in drinking water in Russia and approbation of different methods for cyanotoxins removal from water. Assessment of mutagenic activity of cyanotoxins with use of plant and animal organisms. Study of triggers for expansion of potentially toxigenic cyanobacteria to temperate freshwaters. Yaroslavl State University Lake Nero ) is a shallow, highly eutrophic lake in Yaroslavl Region. The surface area is 57.8 km², a maximum depth - 3.6 m, average -1.6 m . Chl “A” – 90 µg L-1 Total N – 1,8 mg L-1 , Total P – 0,12 mg L-1 Lake Nero Cyanobacteria dominants Micocystis spp., Planktothrix spp., Anabaena spp., Aphanizomenon gracile, Cylindrospermopsis raciborski Genes responsible for cyanotoxin biosynthesis and concentrations of toxins МС СYN PSP mcyE + aoaA+ stxA + 0.04-83 µg/l 0.1-0.4 µg/l data are absent (LC-MS/MS) (LC-MS/MS) Yaroslavl State University Upper Volga Reservoirs • Toxic cyanobacteria producing MCs, CYN, STX and AN-A observed in studied water bodies (PCR analysis). The most numerous were microcystins (95% of studied water samples) and saxitoxins. According PCR analysis: the microcystin producers in studied water bodies were M. aeruginosa, M. viridis, M. flos-aquae, Upper Volga Reservoirs, Uglich M. novacekii, but M. wesenbergii was non-toxic. • 9 variants of MCs, mainly arginine containing MC-LR, RR, YR. (LCMS) Total concentration of detected MCs in different studied water bodies: from 10 ng L-1 (rivers) to 2000-5113 μgL-1 (blooming spots, high trophic water bodies) Komi Republic, Syktyvkar, Russia Institute of Biology of the Komi Science Centre of the Ural Division RAS www.ib.komisc.ru Laboratory of geobotany and comparative floristic, North Flora and Vegetation Department Research interests: Diversity and ecology of Cyanobacteria and algae from the Northeast of Russia (European part), bloom-forming cyanobacteria, water quality Research group on Cyanobacteria: Head of the laboratory Dr. Patova Elena ([email protected]) Sterlyagova Irina, Novakovskaya Irina Komi Republic, Syktyvkar, Russia Komi republic Sampling sites of algalogy studies of cyanobacteria diversity in different kind of ecotopes in European North-East Cyanobacteria are studied in undisturbed tundra and mountain (terrestrial and aquatic) ecosystems As well as in anthropogenically transformed ecosystems Most frequently (high degree of occurrence) the “blooming" of tundra water bodies is caused by : Aphanizomenon flos-aquae Anabaena flos-aquae Microcystis pulverea A. lemmermanii Gloeotrichia echinulata Rivularia planctonica and A. solitaria, A. scheremetievi, A. hassalii, Nostoc linckia, Planktothrix agardhii Anabaena flos-aquae, Aphanizomenon flos-aquae We are also interested in collaboration with different organization to study cyanobacterial toxins, causing blooming in water bodies of European North-East Aphanizomenon Gloeotrichia echinulata flos-aquae Blooming reservoirs is a serious problem during summer Since 2012 we work to create living collection of cyanobacteria and microalgae (http://ib.komisc.ru/sykoa) Institute of the North Industrial Apatity, Murmansk Region, ecological problems, The Barents Euro-Arctic Region Kola Science Center, Russian Academy of Sciences Water ecosystems laboratory www.ineplab22.com Development of a methodology for assessing the current state of freshwater ecosystems Euro-Arctic Region in the context of global and regional environmental change, the theoretical foundations of anthropogenic impact Lapland Biosphere Reserve The head of the laboratory – Dr. Kashulin Nikolay. Imandra Lake Hydrobiological group: Denisov Dmitrii Scientific interests: Algology, bioindication, biodiversity, diatom analyses, paleoecological reconstruction, HABs of the subarctic waters Imandra Lake— the biggest water body of Murmansk Region. Area 876 km2. The maximum depth - 67 m, average depth — 16 m. (67°40′ N 33°00′ E) Limnological studies Fishery , Recreation Use, Drinking water source Anthropogenic stress results in increasing of aquatic toxicity, radical restructuring of the structural and functional characteristics of aquatic communities, changes in trophic status of lakes in 1960- oligotrophic, nowadays - eutrophic, reducing the stability of freshwater ecosystems. HAB in April 4, 2014 Bloom-forming algae species: Anabaena lemmermannii, A. flos-aquae; Biomass of Ceratium hirundinella, monodominant Peridinium goslaviense Anabaena lemmermannii - -1 34mgL Imandra Lake – water body of North region of Russia Since the beginning 2000 HABs and mass mortality of wild fish occurred yearly in Lake Imandra. One of the longest fish-kills lasted from August to September in 2013. Fish-kill in Imandra Lake. July, 2012 Fish-kill in Imandra Lake. August, 2013 Moscow State University, Department of Chemistry, Professor Valery S. Petrosyan ([email protected]) Prevention of cyanotoxincontamination of aquatic ecosystems by means of algacenosis correction with Ch. vulgaris strain IPPAS C-111 The technology is based on the idea of finding the way to decrease the development of cyanobacteria in favor of green algae. The partners of Professor V.S. Petrosyan produce on the industrial scale Chlorella vulgaris IPPAS № С-111, which affects the blooming aquatic ecosystem in a way, that as the result no development of cyanobacteria is taking place and we observe the preferential development of green algae (99%). Moscow State University, Department of Chemistry An example - Barvikha Ponds in Moscow Region For two similar Barvikha ponds the initial phytoplankton composition before starting the experiment was not absolutely identical. The total amount of phytoplankton in the experimental pond before the algolization was 142 880 cells/ml and after the experiment it has decreased till 75 060 cells/ml (mostly the diatomic algae). In the control pond the total amount of phytoplankton vice versa has increased from 356 cells/ml to 13 650 cells/ml. Moscow State University, Department of Chemistry V.S. Petrosyan Chemical Safety and Algolization of Water Reservoirs, International Symposium “The Priority Directions of Revitalization of the Voronezh Water Reservoir”, November 21st, 2012, pp.311-319 the control pond the experimental pond Токсины в биомассе цианобактерий в оз. Сестрорецкий Разлив 2010 2011 2012 MC-LR C49H74N10O12 + + + Demethyl-MC-LR C48H72N10O12 + + + DiDemethyl-MC-LR C47H70N10O12 + ND ND [L-Ser7]MC-LR C48H74N10O13 ND ND + Dehydro-MC-LR C49H76N10O12 + ND ND 6 [D-Glu-OCH3 ] -MC-LR C50H76N10O12 ND + ND MC-RR C49H75N13O12 + + + Demethyl-MC-RR C48H73N13O12 + + + DiDemethyl-MC-RR C47H71N13O12 ND ND + (Dha7)MC-YR C51H70N10O13 + ND ND MC-YR C52H72N10O13 + + + MC-FR C52H72N10O12 ND + AnatoxinThank-a youC10H15NO for your attentionND ! ND +