Antimutagenic effect of neem leaves extract in freshwater fish, Channa punctatus evaluated by cytogenetic tests

Abstract

Neem (Azadirachta indica), an indigenous plant commonly grown in India and its sub-continent is a multipurpose plant well known for its insecticidal and biomedical properties, however, its antimutagenic effects in vertebrate organisms are lacking. The present work is therefore, focused on possible antimutagenic potential of ethanolic extract of neem leaves evaluated on the clastogenicity induced by Pentachlorophenol (PCP) and 2, 4-dichlorophenoxyacetic acid (2,4-D) in freshwater fish, Channa punctatus used as a vertebrate model, by cytogenetic endpoints: chromosome aberration (CA) and micronucleus (MN) test. In the first set of experiment, fish were exposed by medium treatment to a single treatment of each chemical (PCP, 0.6 ppm; 2,4-D, 75 ppm; neem extract, 3 ppm) along with the controls. The chromosome preparations were made after processing kidney cells and micronucleus slides were prepared from peripheral blood at multiple duration (48, 72 and 96 h). PCP and 2,4-D when used alone, induced significant CA and MN in a time dependent manner. Neem extract did not show genotoxic potential in both assays. The maximum frequency of CA were recorded as 18.58% and 15.17%, while frequency of MN reached to 8.08% and 4.62% by PCP and 2,4-D respectively, after 96 h exposure. In the second set of experiment, three concentrations of neem extract (1, 2 and 3 ppm) were run simultaneously with the same concentration of PCP (0.6 ppm) and 2,4-D (75 ppm) for antimutagenicity estimates. In mixed treatment, neem extract significantly reduced the frequency of CA and MN. The reduction in the frequency of CA ranged from 40-75% and 45.4-83.3% and similar values for MN were 40.2-75.3% and 44.1-65.8% for PCP and 2,4-D respectively. Although the reductions were significant but not dependent on concentration and time intervals employed. Results suggested that under present experimental conditions, neem extract exhibit strong antimutagenic activity in this fish model, which could further contribute to study its benefit in humans.