cyanobacteria and cyanotoxins studies in some parts of russian … · 2019-02-20 · cyanobacteria...

Cyanobacteria and cyanotoxins studies

in some parts of Russian Federation

Saint-Petersburg Scientific Research Centre for Ecological

Safety Russian Academy of Sciences

(SRCES RAS)

Ekaterina Chernova , Zoya Zhakovskaya

19-21 February 2015, Seville

Our department - Eco-Chemical studies

Saint-Petersburg

Scientific Research Centre for Ecological Safety

Russian Academy of Sciences

Total neurotoxicity (biosensors)

Anna Pilip

[email protected]

Algological Studies

EkaterinaVoyakina, phD [email protected] Instrumental analysis (LCMS)

– cyanotoxins determination

Iana Russkikh, phD,

[email protected]

Ekaterina Chernova [email protected]

Head of the department

Zoya Zhakovskaya, phD zazhak@hotmail,com

Area–

11,0 km2

Sestroretskij

Razliv Lake

Area–

0,98 km2

Low Suzdal

Lake

Monitoring of HAB of two eutrophic Saint Petersburg

lakes with high recreation load since 2008.

Saint-Petersburg SRCES RAS

Planktothrix agardhii

Microcystis spp. M. aeruginosa

Aphanizomenon flos-aquae

M. viridis

M. wesenbergii

Toxin-producing species

in water bodies of Saint-Petersburg

Saint-Petersburg SRCES RAS

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.

Sestroretskij Razliv Lake a large artificial eutrophic

reservoir primarily used for

recreational purposes.

Surface area – 11,0 km2,

average depth – 1,6 m.

Saint-Petersburg SRECES RAS

HAB in Sestroretskij Razliv Lake, 2010

Aphanizomenon flos-aquae,

Planktothrix agardhii,

Microcystis aeruginosa,

M.wesenbergii, M. viridis

Dominated species:

Detected MC variants:

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

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

Lower Suzdal Lake

Dominated species: Aphanizomenon flos-aquae,

Planktothrix agardhii, Microcystis spp.

The surface area is 0,97 km2,

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).

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)

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.


• Phytoplankton Studies (blue-green algae)

+карты озер

Low Suzdal Lake: Biomass in summer period:________________

Dominated species

Detected concentration of hepatotoxins

МС-LR

Total MCs

MC variants

Scientific Research Centre for Ecological Safety

Russian Academy of Sciences Kaliningrad

* St-Petersburg *

* Rybinsk

Moscow * *Yaroslavl

* Syktyvkar

Apatity

*

* Irkutsk

The Institutions of Russian Federation

running research work on cyanobacteria

Laboratoty for Marine Ecology, Atlantic

Branch of P.P.Shirshov Institute of

Oceanology RAS Kaliningrad

Study of HABs and cyanotoxins

(2010 -2014) has been carried out at the

shallow lagoons of South-Eastern Baltic

and in the marine areas.

Methods: HAB monitoring

(hydrochemistry / biology),

field screening of cyanotoxins(ELISA),

phytoplankton study, experiment.

Elena Ezhova, PhD, Head of Laboratory

([email protected]) (bloom dynamics,

experimental ecotoxicology)

Eugenia Lange, researcher (phytoplankton

composition & dynamics, toxic species)

Maria Maksimova, researcher

(cell culturing, toxin’s biodegradation)

Natalia Molchanova, researcher & Julia Polunina,

PhD, senior researcher (experimental ecotoxicology)

Study of cyanotoxins and its ecological effect

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

screening

http://www.park-kosa.ru/cn_novosti/publikatsii/?ELEMENT_ID=59695






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

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).

Basins of high-productive commercial fishery (bream, herring, eel, whitefish)

Recreational area (national park/reserves, tourism)

MCs concentrations in waters of Curonian Lagoon, Kaliningrad

(LCMS method)

Water sample,

μg L-1

Biomass sample,

μg g-1 (d.w.)

Dominated

species

ΣМСs МС - LR ΣМСs МС - LR

2010 qualitative analysis,

Arg –containing MCs variants were detected

Aph. flos-aquae,

Microcystis spp.

2011 1,0 - 194,8 <LOD- 93,7 304,3 - 3660,7 12,0 – 1714,0 Aph. flos-aquae

Microcystis spp.

2012 0,04 - 18,8

152,6 - 290,5*

0,01 - 3,3

25,0 – 65,5*

1,3 - 658,2 0,1 - 151,5 Aph. flos-aquae,

Microcystis spp

2013 2,2 - 14,9 0,1 - 0,5 25,0 – 4719,0 0,3 – 77,0 Aph. flos-aquae,

Microcystis spp

Curonian Lagoon, Blooming of 2010

* Pelagic zone

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

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

Kaliningrad, Curonian and Vistula Lagoons

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

Rybinskoje

Chebocsarskoje

Gorkovskoje

Volga basin Phytoplankton monitoring of Volga

Reservoirs has run since 1954.

Different trophic status of reservoirs:

Mesotrophic and eutrophic.

280 cyanobacterial species observed

Dominated species:

Aphanizomenon flos-aquae,

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/)

Team:

Scientific Interests:

hepato- and neurotoxigenic cyanobacteria,

ecological role of cyanotoxins,

PCR detection of toxic cyanobacteria,

ecology of shallow lakes, phytoplankton

The head: Babanazarova Olga,

algology, phytoplankton studies

([email protected])

Sidelev Sergey, PCR analysis

([email protected])

Zubishina Alla, ELISA

([email protected])

Applied methods: conventional PCR, real-time PCR, RFLP-

analysis, PCR with individual colonies of toxigenic

cyanobacteria, multiplex PCR, ELISA, fluorescence microscopy

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,

Department of ecology and zoology

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

Genes responsible for cyanotoxin

biosynthesis and concentrations of toxins

МС СYN PSP

mcyE +

0.04-83 µg/l

(LC-MS/MS)

aoaA+

0.1-0.4 µg/l

(LC-MS/MS)

stxA +

data are absent

Cyanobacteria dominants

Micocystis spp., Planktothrix spp.,

Anabaena spp., Aphanizomenon gracile,

Cylindrospermopsis raciborski

Yaroslavl State University

• 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,

M. novacekii, but M. wesenbergii

was non-toxic.

Upper Volga Reservoirs, Uglich

• 9 variants of MCs, mainly arginine

containing MC-LR, RR, YR. (LCMS)

Yaroslavl State University

Upper Volga Reservoirs

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

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

Laboratory of geobotany and

comparative floristic, North

Flora and Vegetation

Department

www.ib.komisc.ru

Sampling sites of algalogy studies of cyanobacteria diversity

in different kind of ecotopes in European North-East

Komi Republic, Syktyvkar, Russia

Komi

republic

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

A. lemmermanii Rivularia planctonica Gloeotrichia echinulata

Microcystis pulverea

and A. solitaria, A. scheremetievi, A. hassalii, Nostoc linckia, Planktothrix agardhii

Blooming reservoirs is a serious

problem during summer

We are also interested in collaboration with different organization to

study cyanobacterial toxins, causing blooming in water bodies of

European North-East

Gloeotrichia echinulata Aphanizomenon

flos-aquae

Anabaena flos-aquae,

Aphanizomenon flos-aquae

Since 2012 we work to create living collection of cyanobacteria

and microalgae (http://ib.komisc.ru/sykoa)

Institute of the North Industrial

ecological problems,

Kola Science Center,

Russian Academy of Sciences

Water ecosystems laboratory

www.ineplab22.com

Apatity, Murmansk Region,

The Barents Euro-Arctic Region

The head of the laboratory –

Dr. Kashulin Nikolay.

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

Hydrobiological group: Denisov Dmitrii

Scientific interests: Algology, bioindication,

biodiversity, diatom analyses, paleoecological

reconstruction, HABs of the subarctic waters

Imandra Lake

Lapland Biosphere Reserve

HAB in April 4, 2014

Biomass of

monodominant

Anabaena lemmermannii -

34mgL-1

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.

Bloom-forming algae species:

Anabaena lemmermannii, A. flos-aquae;

Ceratium hirundinella,

Peridinium goslaviense

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)

Imandra Lake – water body of North region of Russia

Fish-kill in Imandra Lake.

August, 2013

Fish-kill in Imandra Lake.

July, 2012

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.

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, Professor Valery S. Petrosyan

([email protected])

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

Moscow State University, Department of Chemistry

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

[D-Glu-OCH36 ] -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 +

Anatoxin-a C10H15NO ND ND + Thank you for your attention!

cyanobacteria and cyanotoxins studies in some parts of russian … · 2019-02-20 · cyanobacteria...

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