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Molecular detection of microcystin synthetase genes (mcy genes) and semi-quantitative immunological detection of the production of microcystin toxin in vitro-grown pure cultures of cyanobacteria

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Author Affiliations

  • 1Department of Biological Science, Rani Durgavati University, Jabalpur, MP, India
  • 2Department of Biological Science, Rani Durgavati University, Jabalpur, MP, India
  • 3Department of Biological Science, Rani Durgavati University, Jabalpur, MP, India
  • 4Department of Biological Science, Rani Durgavati University, Jabalpur, MP, India

Int. Res. J. Biological Sci., Volume 13, Issue (4), Pages 1-10, November,10 (2024)

Abstract

Laboratory mass cultures were established for cyanobacterial strains M. aeruginosa, O. laetevirens var. minimus, A. fertilissima, P. uncinatum, and S. elongatus. The growth of these cultures was assessed by monitoring turbidity, chlorophyll concentration, and protein content. After an 18-day inoculation period, the maximum growth of pure cultures was observed. Well-developed cultures were concentrated using centrifugation and subsequently lyophilized to preserve them in powdered form. DNA extraction was performed on the lyophilized cultures, resulting in clear DNA bands just below the wells. The quality of the extracted DNA, as determined by the A260/280 ratio, ranged from 1.6 to 1.8. The genes mcyABDE were successfully amplified in M. aeruginosa and O. laetevirens var. minimus, while A. fertilissima and P. uncinatum showed amplification of mcyABD and mcyABE genes, respectively. No amplification was observed in S. elongatus. Using a semi-quantitative ELISA technique, a significant concentration of Microcystin was detected only in Microcystis aeruginosa, at a level of 0.5 ppb, whereas the other cultures produced trace amounts below 0.5 ppb.

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