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Study of anticlastogenic effect of spirulina against arsenic using zebra fish by micronucleus method


Sayantani Chanda1 *, T. V. Ramachandra2, Puspal Dey3 , Madhumita Joarder Mukhopadhay4

1,2, Energy & Wetlands Research Group, Centre for Ecological Sciences [CES],
Indian Institute of Science, Bangalore, Karnataka, 560 012, India,
Web URL: http://ces.iisc.ernet.in/energy; http://ces.iisc.ernet.in/foss,
3,4, Maulana Abul Kalam Azad University of Technology, Kolkata, (W.B.) - India
*Corresponding author:E.mail: isayantani.chanda@gmail.com


Material and Methods

We use zebra fish 3 months old with an average weight of 0.3 g were obtained from a pet shop. This fish is maintained in aquarium with oxygen pumps, thermometer and gives earthworms for feeding. Siphoning is done every day because at the bottom of aquarium dead eggs and uneaten food makes the water hazy. 10 ml stock solution sodium arsenite (30mg dissolved in 100ml) is used. We kept the fishes in each of the two beakers (control and experimental) for 120 hours. At first fishes will taken out of the water and blood was collected from gills. Then 15 slides were taken, saline solution was given on each slide. 1 drop of collected zebra fish blood was given on the slides from the collected gill of the fish through micropipette and slides will smeared. Smear was made from the bloods drops with cover slips. And then dried on the hot plate for 2mins.Slides were stained with freshly prepared 10% giemsa stain for 25mins.Stained slides were washed in normal tap water until the extra stain got washed out and apply tissue paper on the opposite side. Washed slides were left for being air dried.

Red mark denotes arsenic prone district in W.B

Treatment of Danio rerio to Sodium Arsenite (NaAs )

We maintained control (1lt 200ml water) and experimental as described above. 1 litre and 190 ml water + 10 ml stock solution of Sodium Arsenite (30 mg dissolved in 100ml) kept it for 120 hours. After 120 hours we follow the above steps.

Treatment of Danio rerio to Spirulina

Spirulina We maintained control (1lt 200ml water + Spirulina) and experimental as described above. 1 litre and 190 ml water + 10 ml stock solution of Sodium Arsenite(30 mg dissolved in 100ml)+ Spirulina kept it for 120 hours. After 120 hours we follow the same steps. Finally micronucleus was observed from processed blood taken from the treated fish.