EXTRACTION & PURIFICATION OF HEMOLYSIN FROM TOXIN PRODUCING MICROORGANISMS AND ITS EFFECT ON FORENSICALLY IMPORTANT BLOOD AND SALIVA SAMPLES

Abstract

Examination of Blood and saliva samples in forensic investigation is now become crucial part of judiciary as it can be useful for the identification of an individual. Individual can be identified from such biological samples by blood grouping as well as DNA analysis. In the study isolation of microorganisms was done from different surfaces having highest possibility of presence of biological evidence, Gram staining and morphological characterization of microorganisms was done, the extraction of toxin from toxin producing microorganism which was determined by the zone of hemolysis on blood agar media was done by ammonium sulphate and sodium azide method. Purification was done and confirmation of toxin was done by ditch method, TLC & SDS-PAGE. Effect of toxin on blood was examined by Drabkin’s method for determining hemolytic unit. Effect of hemolysin on saliva was examined by starch hydrolysis test as well as loss of an antimicrobial Property of saliva.  As the toxin isolated is hemolysin toxin means it degrades the blood, but in the study we observed the effect of the hemolysin toxin on saliva as well. As mentioned in results blood samples loses its ABO blood grouping activity as it got affected by toxin. Saliva was also highly affected by toxin isolated, as the saliva loses the amylase activity and antimicrobial activity as well. The study is significant in cases where blood and saliva may found as crucial evidence. As RBCs and surface antigens got degraded by hemolysin blood as evidence may be undetectable by presumptive test and may give false results. Saliva samples also become untraceable during the presumptive test as loses its amylase activity.