BENGUET STATE UNIVERSITY, College of As and Sciences Exercise No. 4 EFFECTS OF DISINFECTANTS, ANTISEPTICS AND ANTIMICROBIAL AGENTS Control of microorganisms is essential in the home, industry, and medical fields to prevent and treat diseases and to inhibit the spoilage of foods and other industrial products. Common methods of conirol involve chemical and physical agents that adversely affect microbial structures and functions, thereby producing a microbicidal ormicrobistatic effect. A microbicida! effect is one that kills ihe microbes immediately: a microbistatic effect inhibits the reproductive capacities of the cells and maintains the microbial population at a constant size. Antiseptics are chemical substances used on living tissue that kill or inhibit the growth of vegetative microbial forms. Disinfectants ‘are chemical substances that kill or inhibit the growth of vegetative microbial forms on noniiving materials, while Antibiotics are synthesized and secreted by some true bacteria, actinomycetes, and fungi that destroy or inhibit the growth of other microorganisms. OBJECTIVES: At the end of the activity, the students will be able to: 1. perform the disc-agar diffusion technique for testing antimicrobial activity ‘of common agents. 2. demonstrate the effects of common antiseptics and disinfectants on the growih of gram positive and gram negative bacteria. 3. evaluate the inhibitory effects of certain antibiotics on the growth of bacteria, MATERIALS: To be borrowed from the preparatory room: Department of Biology Y solid plated media (Mueller- Hinton agar or NA) pure cultures of E. coli, B. subtilis and Pseudomonas sp To be brought by students: ¥ cotton swab ¥ filter paper discs ¥ antimicrobial agents 1o be given by your instructor ~ okohol “aataahing Sigua? aay One wodn - Redwmy bg? Tha on Pana Sa a 8BENGUET STATE UNIVERSITY, College of Ats and Sciences Department of Biology PROCEDURI 2 A. DISC-AGAR DIFFUSION TECHNIQUE 1. Obtain three sterile plated media (Mueller- Hinton agar or NA) and inoculate each with the test microorganisms using the swab technique. ‘Swab the entire surface of the plates. 2. Soak ihe filter paper discs into the chemicals to be tested for antimicrobial effects. 3. Transfer the disc on the plated media with the use of sterile forceps. Position the discs at safe distances from each other and press slightly onto the media to avoid movement. Incubate for 24 hours at 24 °C 5. Measure zones of growth inhibition after incubation with the use of a ruler. Zones of growth inhibition are seen as clear zone around the disc. 6. Calculate the microbial index as follows Microbial index= inhibition zone - diameter of disc diameter of disc Draw the pfates after incubation. ‘QUESTIONS FOR RESEARCH 1. Give the actions of each of the antimicrobial agent tested. 2. Interpret the effectivity of the antimicrobials used based on the results of the activity. 3. What ore some factors that affect the actions of chemical ‘antimicrobial agents? Explain.