RESUMEN
Objective: In this study, albino mice were injected with a sub-lethal dosage of purified wasp Ropalidia Marginata venom toxins to assess the effectiveness of polyclonal anti-venom antibodies.Methods: To neutralize the toxic effects, polyclonal antibodies were generated by immunizing albino mice. The antibody underwent partial purification using ammonium sulphate treatment and octanoic acid precipitation. To detect the presence of antibodies in the antiserum, an immunodouble diffusion test was conducted using Ouchterlony's method (1962). This involved allowing both antigens and antibodies to diffuse radially towards each other from their respective wells. When they reached an equivalence zone, a precipitation complex of antigen and antibody became visible as a concentric band, indicating the development of the combination. To quantitatively determine the amount of antibodies in the antiserum, the equivalency zone approach was used.Results: Experimental mice were injected with a combination containing 400, 800, and 1200 µg of pure antibody, which had been treated serum biomolecules, including metabolic enzymes, completely reversed in the experimental with 40% of the LD50 of wasp venom the elevated serum parameters were glucose, pyruvic acid, lipid, protein and free amino acid, reached to normal (100%) in the treated with 40% of LD50 of the venom and polyclonal treated after 6 h of administration. Anti-serum treatment also successfully normalized the alteration in serum enzyme just after 4h.Similarly, anti-serum treatment also successfully normalized the alteration in serum enzyme just after 4h treated with 40% of LD50 of the venom. Serum ACP content was obtained as 125.35% after 40% of LD50 venom injection, which was get normalized up to 102.81% after 4 h of the anti-venom treatment. Serum ALP content of 114.8% elevation was reversed back to 102.40% after anti-venom treatment. The GPT level significantly reversed up to 102.5%, while it was 130% in the venom-treated mice. A complete reversal was obtained in GPT level, which was obtained as 104.54% in the venom-treated animal. Similarly, LDH which was elevated up to 112.45 % in venom-injected mice was successfully reversed up to 100.25% after anti-venom treatment. Similarly, Ache concentration was fully recovered after anti-venom treatment 6 h, all animals (group B-E) that had received 40% of the LD50 of venom treated with pure antiserum.Conclusion: The venom-injected group showed a complete restoration of serum protein, free amino acid, uric acid, cholesterol, pyruvic acid, total lipid, and glucose level in experimental mice.
RESUMEN
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.
RESUMEN
Absorption and transport of 3H cholesterol from the midgut to hemolymph and other tissues was studied in the locusts Schistocerca gregaria and Locusta migratoria. S. gregaria are able to absorb dietary cholesterol in the midgut and release into the hemolymph in vivo and into the incubation medium in virto. Certain proteins of midgut origin are involved in the absorption and release of cholesterol. The proteins designated as cholesterol binding proteins (CBP's) were fractionated by gel filtration chromatography using Sepharose CL-6B-200 column. Presence of a protein and its binding with cholesterol is confirmed by TCA precipitation after subsequent incubation of midgut in the incubation medium. Cholesterol binding with the proteins was also confirmed in native polyacrylamide gel electrophoresis. Biosynthesis of this protein takes place in the midgut which is inhibited by a protein synthesis inhibitor, cycloheximide. It also inhibits absorption and release of cholesterol from the midgut. The cholesterol binding activity was associated with a peak containing proteins ranging from molecular weights of 17-32 kDa in SDS-PAGE gels. Treatment of midgut with cycloheximide resulted in reduced cholesterol binding activity. Dilipidation of mucin and transport in presence of bile salts yielded a higher cholesterol binding activity. Although the absorption and release of cholesterol was observed in the hemolymph of both sexes, the ovary exhibited higher cholesterol binding as compared to testis.