Enterobacter aerogenes (E. aerogenes) has been commonly described as a versatile opportunistic pathogen in hospital infections. The aim of the present work was to evaluate the impact of biofield treatment on E. aerogenes for its phenotypic and genotypic characteristics. E. aerogenes bearing ATCC 13048 (American Type Culture Collection) was procured from Bangalore Genei, in sealed pack and divided into control and treated groups. Treated group was subjected to Mr. Trivedi’s biofield treatment and analyzed for antimicrobial susceptibility, minimum inhibitory concentration (MIC), biochemical reactions, and biotype using automated MicroScan Walk-Away® system. In addition, treated group of E. aerogenes was evaluated for DNA polymorphism by Random Amplified Polymorphic DNA (RAPD) and 16S rDNA sequencing to establish the phylogenetic relationship of E. aerogenes with different closely related bacterial species. Antimicrobial susceptibility results showed an alteration of 14.28% among twenty-eight tested antimicrobials. Similarly, 15.65% tested antimicrobials showed an alteration in MIC values. Chloramphenicol showed improved sensitivity i.e. resistant to susceptible after biofield treatment, with the support of decreased MIC by two folds (i.e. >16 to ≤8 µg/mL). Norfloxacin also showed decrease MIC by two folds (i.e. 8 to ≤4 µg/mL) as compared to control. Biofield treatment showed an impact on biochemical reactions (9.09%) followed by a change in biotype number (7770 5272) in treated group with respect to control (7770 5372). Using RAPD analysis, sample showed an average range of 4 to 42% of polymorphism, while 16S rDNA study showed that treated sample was detected as Kluyvera cryocrescens (GenBank Accession Number: AM184245) with 97% identity of gene sequencing data, which was nearest homolog species to Enterobacter aerogenes strain: C1111 (Accession No. AB244467). These results suggest that Mr. Trivedi’s unique biofield treatment can alter the antimicrobial sensitivity pattern, thus it can be used as alternate energy medicine in future.