Toxicology International (Formerly Indian Journal of Toxicology)

Preety Bhinder , Asha Chaudhry

Abstract

Objectives: In this study we have evaluated the genotoxic potential of pesticides acephate and profenofos by polymerase chain reaction‑restriction fragment length polymorphism (PCR‑RFLP) assay with the mosquito Culex quinquefasciatus taken as experimental model. Material and Methods: Second instar larvae were treated with LC₂₀ of each pesticide for 24 h and induced mutations in the sequence of mitochondrial 16S rRNA gene were studied from restriction patterns generated with PacI and PsiI restriction endonucleases. Results: Variations in the number and size of digested fragments were recorded from treated individuals compared with controls showing that the restriction enzymes created a cut at different locations. In addition, sequences of the 16S gene from control and treated individuals were also used to confirm the RFLP patterns. From the sequence alignment data, it was found that mutations caused the destruction and generation of restriction sites in the gene sequence of treated individuals. Conclusion: This study indicates that both the pesticides had significant potential to induce mutations in the 16S gene of Culex quinquefasciatus.

Keywords

Acephate, culex quinquefasciatus, genotoxicity, pcr‑rflp, profenofos

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Preety Bhinder , Asha Chaudhry

Abstract

Objectives: In this study we have evaluated the mutagenicity of organophosphate pesticides acephate, chlorpyrifos, and profenofos using polymerase chain reaction‑restriction fragment length polymorphism (PCR‑RFLP) assay with the mosquito Culex quinquefasciatus taken as an experimental model. Materials and Methods: Second instar larvae were treated with LC20 of each pesticide for 24 h and mutations induced in the sequence of mitochondrial COII gene (690bp) were studied from restriction patterns generated with AluI, PacI, and PsiI restriction endonucleases. Results: Variations in the number and size of digested fragments were recorded from treated individuals compared with controls showing that the restriction enzymes created a cut at different locations. In addition, sequences of COII gene from control and treated individuals were also used to confirm the RFLP patterns. From the sequence alignment data, it was found that mutations caused the destruction and generation of restriction sites in the gene sequence of treated individuals. Conclusion: This study indicates that all the three pesticides had potential to induce mutations in the normal sequence of COII gene and also advocates the use of PCR‑RFLP assay as an efficient, rapid, and sensitive technique to detect mutagenicity of pesticides

Keywords

Acephate, chlorpyrifos, Cx. quinquefasciatus, mutagenicity, PCR‑RFLP, profenofos

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Preety Bhinder ¹, Asha Chaudhry ¹, Bhupinder Barna ¹, Satvinderjeet Kaur ¹

Affiliations
1 Department of Zoology, Punjab University, Chandigarh - 160 014, IN

Abstract

The present article deals with the polymerase chain reaction (PCR)-based genotoxicity evaluation of neonicotinoid pesticides, imidacloprid and thiamethoxam, by using the genome of a mosquito Anopheles stephensi taken as an experimental model. After treatment of the second instar larvae with LC₂₀ of the pesticides for 24 h, the induced nucleotide sequence variations in the internal transcribed spacer 2 (ITS2) of freshly hatched unfed control and treated individuals was studied from the sequence alignment data and the mutations in the form of insertion, deletion and substitution of bases were recorded. Measurable differences, indicative of the genetic damage due to imidacloprid and thiamethoxam were observed when ITS2 sequences of control and treated individuals were compared. It was found that imidacloprid-treated individual had 8 deletions, 29 insertions, 18 transitions and 33 transversions, whereas thiamethoxam-treated individual had 10 deletions, 8 insertions, 47 transitions and 68 transversions.

Keywords

Anopheles stephensi, Genotoxicity, imidacloprid, thiamethoxam

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