CLS 402:

Lab #3:
**Work in pairs for each lab practice (1 set of plates for each pair) but please record individual answers for grading. **Lab safety: Assume all specimens are infectious!! Wash hands when leaving the laboratory or if they come in contact with infectious surface, bacteria or specimen. Wear gloves when appropriate. NO hand to mouth or hand to eye contact. Disinfect surfaces before leaving the laboratory or if spills or splatters occur. If in doubt, wash your hands or clean the surface.

1. Read out urine, respiratory, wound, stool and positive blood culture (s). Record results on provided sheets. Use the guide for organism identification, textbooks, provided materials and the instructor for assistance if needed. 2. Read API and assign organism ID. 3. Set up Antimicrobial Susceptibility testing on an organism after consultation with instructor (see handout). 4. Inoculate a screen plate after instructor explanation. 5. Read student slide set.

Lab #4:
1. Complete readouts for the week. All cultures should be complete. If not, consult instructor. 2. Read and record AST. 3. Discuss anaerobic culture methods and isolation. 4. Complete student slide set.

Lab #5:
1. Streak plates for grading for Practical/Final 2. Culture specimens. 3. Review for Practical. Review Questions (5pts/question): 1. Discuss the utilization and concept of screen plates. Screen plates are designed in order to help identify certain bacteria. Certain types of screen plates will contain specific components or nutrients that help to identify the targeted bacteria. In some cases, certain color changes will take place when the targeted bacterium is present on the plate.

2. Outline the Kirby Bauer susceptibility testing method. Kirby-Bauer antibiotic testing is a test which uses antibiotic disks to test whether particular bacteria are susceptible to specific antibiotics. A known quantity of bacteria is grown on agar plates in the presence of thin wafers containing relevant antibiotics. If the bacteria are susceptible to a particular antibiotic, an area of clearing surrounds the wafer where bacteria are not capable of growing (called a zone of inhibition). This along with the rate of antibiotic diffusion are used to estimate the bacteria's sensitivity to that particular antibiotic. In general, larger zones correlate with smaller MIC of antibiotic for that bacteria. This information can be used to choose appropriate antibiotics to combat a particular infection. 3. Discuss anaerobic culture methods. In order to culture an anaerobic specimen, it is important to avoid any oxygen contamination. After the sample has been collected, immediate storage in an anaerobic jar, chamber, or solution is required. This will ensure that the environment remains without oxygen and survival of the possible anaerobic bacteria. 4. Discuss contamination vs. true infection in the case of a positive blood culture. What conditions are met in each situation? With any blood culture, two samples should be taken from two different body sites. In the case of a contaminated sample, one sample will contain bacteria A, while the second sample lacks bacteria A. In the case of a true infection, both samples will contain bacteria A.