PRODUCTION OF POLYCLONAL ANTIBODIES AGAINST INDIAN CATTLE TICK RHIPICEPHALUS MICROPLUS SALIVA TOXINS AND ITS EFFICACY IN REVERSAL OF TOXIC EFFECTS IN ALBINO MICE

Abstract

Objective: This study focuses on the generation of polyclonal antibodies against tick saliva toxins and its use to reverse the toxic effects in albino mice. Methods: Polyclonal antibodies were generated by immunizing albino mice were immunized with saliva toxins mixed with incomplete Freund’s adjuvant. Experimental mice were treated with antiserum (polyclonal antibodies) and pre-incubated with tick saliva toxins in five different groups for observation of reversal of toxic effects, i.e. levels of bio-molecules and enzymes. For detection of polyclonal antibodies in the antiserum immune double diffusion (IDD) test of Ouchterlony was followed. Results: By employing a step-by-step octanoic acid and ammonium sulphate precipitation process, IgG antibodies were separated from antiserum. A crescent band and precipitation band was obtained due to the interaction of antigen and antibodies in wet agarose gels (1%). When these antibodies were injected in albino mice, these have been successfully reversed the levels of acid phosphatase (ACP), alkaline phosphatase (ALP), glutamate pyruvate transaminase (GPT), glutamate oxaloacetate transaminase (GOT), lactic dehydrogenase (LDH) and acetylcholinesterase (AchE). Alkaline phosphate levels in the serum of albino mice injected with polyclonal antibodies were found to be 122.64%, 107.849%, and 104.71%, respectively. Glutamate pyruvate transaminase (GPT) has been reversed in mice treated with polyclonal antibodies up to 94.59%, 86.48% and 78.37% in the serum, while it was found to be 116.21% at 40% of 24-h LD50 dose in comparison to control respectively. Similarly, level of lactic dehydrogenase was restored and found i.e. 104.55%, 103.82%, and 102.20% in the serum of albino mice. Respectively, in comparison to control, while mice injected with 40% of 24-h LD50 of the purified saliva toxins demonstrated 117.20% of lactic dehydrogenase (LDH) level in comparison to control. Conclusion: Polyclonal antibodies administered for serotherapy reversed the toxic effects and all biochemical parameters become normal after 6 h of treatment in albino mice in comparison to control.