Bacillary dysentery and acute gastroenteritis caused by infection of Shigella species are major public health burden in India and its neighboring countries. Emergence of antimicrobial resistance threatens to render current treatments ineffective. The current study was attempted to investigate the effect of biofield treatment on Shigella boydii (S. boydii) with respect of antimicrobial susceptibility assay, biochemical characteristics and biotyping. The American Type Culture Collection (ATCC 9207) strain of S. boydii was used in this experiment. The study was conducted in revived and lyophilized state of S. boydii. Both revived (Group; Gr. II) and lyophilized (Gr. III) strain of S. boydii were subjected to Mr. Trivedi’s biofield treatment. Gr. II was assessed on day 5 and day 10, while Gr. III on day 10 with respect to control (Gr. I). Sensitivity pattern of amoxicillin/k-clavulanate was improved from intermediate (I) to susceptible (S) with correspond to MIC value was also reduced by two folds (16/8 to ≤ 8/4 µg/mL) in both the treated groups as compared to control. The antimicrobial susceptibility of S. boydii showed 15% alteration in Gr. II on day 5, while significant (40%) alteration was found on day 10 as compared to control. The MIC values of antimicrobials for S. boydii also showed 12.50% alteration in Gr. II on day 5 while, significant alteration (59.38%) of minimum inhibitory concentration (MIC) value was found in Gr. II on day 10 as compared to control. It was observed that overall 69.70% biochemical reactions were changed in which 66.67% alteration was found in Gr. II on day 10 with respect to control. Moreover, biotype numbers were changed in all the treated groups without alteration of organism as compared to control. These results suggested that biofield treatment had significant impact on S. boydii in Gr. II on day 10 with respect to antimicrobial susceptibility, MIC and biochemical reactions pattern.