Antiseptics and disinfectants are antimicrobial agents, referred as biocides, well known as the first line of defense against diseases. They are chemical agents worldwide used in hospitals, healthcare settings, household and industry to destroy or inhibit microbial growth. The present study, investigates bacterial contamination of in- use antiseptics (N=40), disinfectants (N=40), unused antiseptics and disinfectants (N=10) as control and closures (N=10). Samples were collected randomly from various places of the community at Amman City/Jordan. Status of collected antiseptic and disinfectants, behavior and practices of people using them were assessed to understand factors contributing to their contamination, through direct interviewing and filling questionnaire sheets. Incidence of contaminations of antiseptics and disinfectants according to time extent of usage and the active agent was determined. In- Use test was adopted to detect contamination of collected samples followed by culturing onto various types of selective and differential media to identify types of contaminating bacteria. Cell morphology was examined microscopically and characteristics of grown colonies and their reactions in different media were examined for presumptive identification of bacteria at the generic level. Biochemical tests in addition to three different sets of API kit systems were utilized for the identification at the species level. Seventy six bacterial isolates were obtained; the most predominant was Bacillus spp. (65.8%), of which six species were identified, the most prevalent were: B. lentus (32%), B. cereus (26%) and B. subtilis (20%). The second most prevalent bacteria was Staphylococcus spp. (26.3%) of which CoNs and CoPs constitute (50% and 45% respectively), in addition to Acinetobacter calcoaceticus (6.6%), and Serratia marcescens. (1.3%). Identified species were examined for their susceptibility to various antibiotics. Multiple Resistant to up to four antibiotics was detected by B. lentus. Heteroresistance was presented by three Bacillus spp. including B. lentus. S. epidermidis was the only species among identified Staphylococci displaying heteroresistance towards two antibiotics. Biocide susceptibility of most prevalent bacterial species was determined toward four antiseptics and disinfectants including: 10% Iodine 0.13% Benzalkonium chloride, 70% Alcohol and 80% Alcohol under clean and dirty conditions. Dirty condition exerted marked effect in reducing biocides efficacy, both 10%iodine and 0.13% benzalkonium chloride demonstrated the highest activity among other tested biocides. Nevertheless, B. lentus was found completely resistant to all biocides under both clean and dirty conditions. Heteroresistance was displayed only by S. epidermidis in the presence of 70% alcohol in both clean and dirty conditions.