Clinicians now recognize that both aerobic and anaerobic microorganisms have the ability to degrade or damage host tissue at a wound site through the production of a variety of enzymes and toxins. Silver-containing dressings offer one method for controlling this polymicrobial wound bioburden, and research efforts are currently ongoing to determine their efficacy against aerobic, anaerobic, and antibiotic-resistant microorganisms. The current study aimed to determine the antimicrobial activity of a new silver-containing Hydrofiber dressing (AQUACEL Ag) on both aerobic and anaerobic microorganisms, using the zone-of-inhibition method. This method provides a measure of the ability of the dressing to make available a sufficient concentration of silver to have an antimicrobial effect. To some extent this test mimics the clinical use of the dressing and predicts its microbicidal activity at the wound-dressing interface. The results show that the silver-containing dressing makes silver available at a dressing-agar interface at a concentration that is effective against a broad range of aerobic, anaerobic, and antibiotic-resistant microorganisms. In the context of wound healing, the results showing antimicrobial activity against antibiotic-resistant microorganisms are particularly important, as the control and eradication of these organisms is a major concern within the health care profession.