MATERIALS AND METHODSZebrafish care

MATERIALS AND METHODS
Zebrafish care, maintenance and breeding
Wild type zebrafish (Danio rerio) were
obtained from a pet shop in Anantapur and
were bred for two generations in our aquarium
facility by following the conditions described
by Westerfield (1995). Healthy fish of third
generation were only used for the present experimentations.
Fifty fish of third generation
were housed in each 100L glass aquaria with
continuous aeration at a temperature of 27 ±
1°C and 13:11 hr light:dark photoperiod. Fish
were fed twice a day with alternating diet of
freshly hatched brine shrimp (sanders brine
shrimp co, utah) in the morning and dry flake
food (tetra brand) in the evening.
Toxicant tested – preparation of stock solution
Technical grade deltamethrin (S)-α-
cyano-3-phenoxybenzyl (1R, 3R)-3-(2, 2-
dibromovinyl)-2, 2-dimethylcyclopropanecarboxylate-
99.8% pure) was obtained from
sigma-aldrich (Laborchemikalien GmbH D-
30918). The stock solution was prepared by
dissolving 10mg of deltamethrin in 2ml of absolute
ethanol (99% pure) and stored at 4°C.
From this daily requirement are taken for daily
renewal of the test solution in the aquaria.
Exposure strategy
Mature adult zebrafish in equal ratio
(n=8) of male and female from our aquarium
stock were kept in 20L glass aquaria with continuous
aeration for ten days. Later 2μg/L and
5μg/L of DM was added. Simultaneously control
and solvent control (addition of same
quantity of solvent used to dissolve DM) fish
were maintained separately. About 50% of
water in the aquaria was removed daily and
filled with fresh water; however concentrations
of DM and solvent at desired levels were
maintained by fresh additions accordingly. After
the stipulated time i.e after 6 days of exposure
fish were anaesthetized with methanesulfonate
salt (MS-222; Sigma) and then blood
and gonads were collected from both male and
female fish for conducting the experiments
mentioned below. The experimental protocol
carried out was approved by the Institutional
Animal Ethics Committee of Sri KrishnadevarayaUnviersity,
Anantapur, India.

Hasil (Bahasa Indonesia) 1:

MATERIALS AND METHODSZebrafish care, maintenance and breedingWild type zebrafish (Danio rerio) wereobtained from a pet shop in Anantapur andwere bred for two generations in our aquariumfacility by following the conditions describedby Westerfield (1995). Healthy fish of thirdgeneration were only used for the present experimentations.Fifty fish of third generationwere housed in each 100L glass aquaria withcontinuous aeration at a temperature of 27 ±1°C and 13:11 hr light:dark photoperiod. Fishwere fed twice a day with alternating diet offreshly hatched brine shrimp (sanders brineshrimp co, utah) in the morning and dry flakefood (tetra brand) in the evening.Toxicant tested – preparation of stock solutionTechnical grade deltamethrin (S)-α-cyano-3-phenoxybenzyl (1R, 3R)-3-(2, 2-dibromovinyl)-2, 2-dimethylcyclopropanecarboxylate-99.8% pure) was obtained fromsigma-aldrich (Laborchemikalien GmbH D-30918). The stock solution was prepared bydissolving 10mg of deltamethrin in 2ml of absoluteethanol (99% pure) and stored at 4°C.From this daily requirement are taken for dailyrenewal of the test solution in the aquaria.Exposure strategyMature adult zebrafish in equal ratio(n=8) of male and female from our aquariumstock were kept in 20L glass aquaria with continuousaeration for ten days. Later 2μg/L and5μg/L of DM was added. Simultaneously controland solvent control (addition of samequantity of solvent used to dissolve DM) fishtetap dipertahankan secara terpisah. Sekitar 50%Air di aquaria dihapus setiap hari dandiisi dengan air segar; Namun konsentrasiDM dan pelarut pada tingkat yang diinginkan yangdikelola oleh segar penambahan sesuai. Setelahyaitu waktu ditetapkan setelah 6 hari eksposurikan yang anaesthetized dengan methanesulfonategaram (MS-222; Sigma) dan kemudian darahdan gonad dikumpulkan dari kedua laki-laki danPerempuan ikan untuk melakukan eksperimendisebutkan di bawah ini. Protokol eksperimentalmembawa out disetujui oleh kelembagaanHewan etika Komite Sri KrishnadevarayaUnviersity,Anantapur, India.