In vitro assessment of carbendazim induced immunotoxicity

The present investigation aims at evaluating the carbendazim for deleterious effects and induction of apoptosis in avian lymphocytes. For this purpose one batch of splenic avian lymphocytes was treated with various concentrations (NOEL/10², NOEL/10³, NOEL/10⁴, NOEL/10⁵, NOEL/10⁶ and NOEL/10⁷) of carbendazim for 30, 60 and 90 min and another batch of cells was kept as untreated control. The cells were collected for detection of ultrastructural alterations by transmission and scanning electron microscopy. Immunoperoxidase staining was performed for demonstration of phosphatidylserine translocation using Annexin-V biotin conjugate. DNA was isolated from treated and control cells and agarose gel electrophoresis was performed for DNA ladder pattern. Lymphocyte proliferation assay was conducted to assess the effect of carbendazim on T- and B-cell blastogenesis. In carbendazim treated cells, ultrastructural changes included pyknosis, karyorrhexis of chromatin material which marginated on nuclear membrane and shrinkage of cell. The SEM findings revealed the formation of blebs of various size and shape on the surface of cells. The translocation of phosphatidylserine was demonstrated on cell membrane using immunoperoxidase technique. Ladder pattern was observed in agarose gel electrophoresis with smearing of oligonucleotides in comparison to controls and standard DNA markers, which is highly suggestive of apoptosis in avian lymphocytes due to carbendazim exposure. It can be concluded from the present study that the carbendazim induces apoptosis in avian lymphocytes which constitutes the first report of its kind.