International Research Journa Vol. 6(2), 22-29, February (201 International Science Community Associat Distribution of microcysti phytoplankton and prod from E Prashant Cha Cyanobacterial Research Laboratory, Departme Avai Received 29 th Novem Abstract The important value index, which is sum cyanobacterial genera present in phytopla Madhya Pradesh. All such scum/mat samp Anabaena, Nostoc, Phormidium and Sp cyanobacterial scum/mats samples indica ubiquitous presence, mcy genes displayed together. Present study showed amplifica dissolved microcystin content in the wat ELISA. About 30% water bodies contain presence of the toxin in a range of 0.5-3.0 use of water for drinking and recreational Keywords: Biodiversity, Agarose gel elec Introduction Microcystin is produced by bloom fo cyanobacteria 1,2 . Approximately 80 diffe microcystins have been identified, being pro cyanobacterial genera. The commonest are leucine-arginine (LR), –arginine-arginine, (R arginine (YR) variants 3 . Microcystins are toxi lead to impaired liver function and even fata are synthesized by non ribosomal peptide/pol multi-enzyme complex 4 . Cyanobacteria fall Microcystis, Anabaena and Planktothrix potentially known to produce microcy Phormidium and Nostoc are also considere producing genera, they are rarely associated bloom formation 7-9 . Microcystins are cyclic which ADDA (3-amino-9-methoxy 2 phenyldeca-4,6-dienoic acid) and D-Glu i inhibit protein phosphatases and manif responses 10,11 . Almost all toxigenic filament occurring in planktonic or benthos forms suc Phormidium, Anabaena, and Nostoc produce water system, and since the planktonic cyanob undergo lysis and decomposition 12 , microcyst water. Consequently, the toxin threatens sustainability of aquatic system, affecting q al of Biological Sciences ___________________________ 17) tion in synthetase genes in filamentous duction of microcystin in water sam Eastern Madhya Pradesh, India aturvedi, Trashi Singh and Suvendra Nath Bagchi * ent of Post Graduate Studies and Research in Biological Science Jabalpur, MP, India [email protected]ilable online at: www.isca.in, www.isca.me mber 2016, revised 30 th January 2017, accepted 5 th February 201 m of relative abundance, biovolume and frequency, wa ankton scum/mat material in ten water bodies located in ples were dominated by cyanobacteria mostly belonging t pirulina. The microcystin synthetase genes (mcyABD ating that toxic genotypes of cyanobacteria constituted d quite a patchy distribution pattern, and rarely all the cation of mcyA (80%), mcyB (60%), mcyD (50%) and ters harbouring cyanobacterial populations was determ ined the free microcystins below 0.5 ppb, whereas the 0 ppb. These values are well below 1 μg L -1 microcystin, l purposes according to WHO guidelines. ctrophoresis, ELISA, Microcystin, PCR amplification. orming toxigenic erent variants of oduce by different e the microcystin– RR) and –tyrosine- ic in nature, which ality 2 . Microcystins lyketide synthatase ling under genera (Oscillatoria) are ystins 5,6 . Though ed as microcystins with profuse toxic c heptapeptides in 2,6,8-trimethyl-10- interact with and fest toxicological tous cyanobacteria ch as Oscillatoria, microcystin in the bacteria frequently tins are released in the ecology and quality of drinking water, irrigation, fishing, recreation Moreover, microcystin accumulati many diseases in human and ani investigated the diversity and cyanobacteria in different water presence of mcyABDE genes by microcystin concentration was also qualitative ELISA technique. Materials and methods Chemicals: All general purpose ch HiMedia (India) and Sigma-Aldri PCR amplification were purchased Pvt. Ltd. (India) and ELISA kit p Inc. (USA). Survey of study area and samplin ponds, lakes and man-made reservo and Dindori districts (79ºE – 81ºE latitude) were surveyed, and durin cyanobacterial samples in the form collected in wide mouth plastic bot box. The morphological features genus level were examined accord by Desikachary 16 at 10 X and 40 X lenses (Olympus, Optical Corporati __________ ISSN 2278-3202 Int. Res. J. Biological Sci. 22 s cyanobacterial mples collected e, Rani Durgavati University, 17 as calculated for different n three districts of Eastern to the genera Oscillatoria, DE) were detected in all d the populations. Despite e four genes were present mcyE (80%) genes. The mined by semi-quantitative e remaining ones showed , a benchmark set for safe n and aquatic food products. ing into food chain leads to imals 13-15 . In this paper we dominance of filamentous bodies and determined the y PCR amplification. Free o analyzed in waters by semi- hemicals were procured from ich (USA). The primers for from Imperial Life Sciences purchase from Enviro Logix ng locations: Ten permanent oirs located in Jabalpur, Seoni E longitude and 22ºN – 24ºN ng March 2014 - June 2015 m of floating scum/mats were ttles and brought to lab in ice for identification up to the ding to the keys as described X magnifications of objective ion Ltd, Japan).
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International Research Journal of
Vol. 6(2), 22-29, February (201
International Science Community Association
Distribution of microcystin synthetase genes in filamentous cyanobacterial
phytoplankton and production of microcystin in water samples collected
from Eastern Madhya Pradesh, IndiaPrashant Chaturvedi, Trashi Singh and Suvendra Nath Bagchi
Cyanobacterial Research Laboratory, Department of Post Graduate Studies and Research in Biological Science, Rani Durgavati Un
Available online at: Received 29th November
Abstract
The important value index, which is sum of relative abundance, biovolume
cyanobacterial genera present in phytoplankton scum/mat material in ten water bodies located in three districts of Eastern
Madhya Pradesh. All such scum/mat samples were dominated by cyanobacteria mostly belongi
Anabaena, Nostoc, Phormidium and Spirulina
cyanobacterial scum/mats samples indicating that toxic genotypes of cyanobacteria constituted the populations. Despit
ubiquitous presence, mcy genes displayed quite a patchy distribution pattern, and rarely all the four genes were present
together. Present study showed amplification of
dissolved microcystin content in the waters harbouring cyanobacterial populations was determined by semi
ELISA. About 30% water bodies contained the free microcystins below 0.5 ppb, whereas the remaining ones showed
presence of the toxin in a range of 0.5-3.0 ppb. Th
use of water for drinking and recreational purposes according to WHO guidelines.
Keywords: Biodiversity, Agarose gel electrophoresis, ELISA, Microcystin
Introduction
Microcystin is produced by bloom forming toxigenic
cyanobacteria1,2
. Approximately 80 different variants of
microcystins have been identified, being produce by different
cyanobacterial genera. The commonest are the microcystin
leucine-arginine (LR), –arginine-arginine, (RR) and
arginine (YR) variants3. Microcystins are toxic in nature, which
lead to impaired liver function and even fatality
are synthesized by non ribosomal peptide/polyketide synthatase
multi-enzyme complex4. Cyanobacteria falling under genera
Microcystis, Anabaena and Planktothrix
potentially known to produce microcystins
Phormidium and Nostoc are also considered as
producing genera, they are rarely associated with profuse toxic
bloom formation7-9
. Microcystins are cyclic heptapeptides in
which ADDA (3-amino-9-methoxy 2,6,8
phenyldeca-4,6-dienoic acid) and D-Glu interact with and
inhibit protein phosphatases and manifest toxicological
responses10,11
. Almost all toxigenic filamentous cyanobacteria
occurring in planktonic or benthos forms such as
Phormidium, Anabaena, and Nostoc produce microcystin in the
water system, and since the planktonic cyanobacteria frequently
undergo lysis and decomposition12
, microcystins are released in
water. Consequently, the toxin threatens the ecology and
sustainability of aquatic system, affecting quality of drinking
Journal of Biological Sciences ___________________________
(2017)
Association
Distribution of microcystin synthetase genes in filamentous cyanobacterial
phytoplankton and production of microcystin in water samples collected
from Eastern Madhya Pradesh, India Chaturvedi, Trashi Singh and Suvendra Nath Bagchi
*
Cyanobacterial Research Laboratory, Department of Post Graduate Studies and Research in Biological Science, Rani Durgavati Un