SU 2012

BIO 260L Microbiology Lab Practical Study Guide

An individual who is well prepared for Lab Practical 2 will be able to elaborate on each of the following learning objectives. 1) Know safety features of the laboratory. 2) Known all of the parts of the microscope discussed in lab 3) Describe examples of aseptic techniques used in lab. 4) Know what occurs during a Gram reaction for each step in both gram positive and gram negative. a. Know the difference between Gram positive, negative and variable b. Know why a Gram- organism stains negative c. Know what a Gram + and organism will look like during each step of the staining process 5) Discuss the morphological characteristics of the following bacteria: Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus pyogenes, Streptococcus pneumoniae, Bacillus subtilis, E. coli, K. pneumoniae, Salmonella spp., and Pseudomonas aeruginosa. a. Describe and identify the cellular morphology arrangements b. Describe and identify the Gram stain reaction c. List the location on the body where these bacteria are most likely to be found as normal flora and pathogens. 6) Know which genera of bacteria produce endospores and the appearance of spores vs. vegetative cells on an endospore stain. Know the primary and counter stains. 7) Know how to calculate the initial concentration of a sample if you are given the CFUs on a spread plate and dilution factor from a dilution series. 8) Known the purpose of a streak plate and be able to demonstrate how to prepare a streak plate 9) Understand the purpose and results of the agar plate media used in class (TSAVJA, BAP, MSA/VJA, MacKonkeys) a. Describe each media type as simple, enriched, selective, and/or differential b. If the media is selective, know what it selects for and what it inhibits c. If the media is differential, know the appearance of the differentiated organisms d. Known the key ingredients and pH indicator of each type of media e. Describe the biochemistry of the results, (for example, describe why Staphylococcus aureus turns MSA/VJA yellow). 1

SU 2012 f. List which groups of bacteria you would most likely use each media type to identify 10) Identify the 3 types of hemolysis on a BAP plate. Know the hemolysis characteristics of the following organisms and how they appear on BAP a. Streptococcus pyogenes b. Stapylococcus aureus c. Staphylococcus epidermidis d. Streptococcus pneumoniae e. Streptococcus agalactiae 11) Understand the purpose and results of the tubed media used in class (Phenol Red Broth, Citrate, Urea, SIM, TSI) a. Know the purpose of each test b. Know the key ingredients and pH indicator of each test c. Describe the biochemistry behind each color change for each test 12) Understand the purpose and results of the catalase, oxidase, coagulase, and Bacitracin tests a. Know the purpose of each test and which organism is identified by each test b. Know how each test is performed c. Identify the appearance of a positive and a negative result 13) Understand the results of an Enterotube II test a. Identify if a reaction is positive or negative when provided a results key b. Identify an unknown organism when given an organism key 14) Compare the effectiveness of disinfectants if given plates of bacteria treated with various disinfectants. 15) Compare antibiotic sensitivity vs. resistance by measuring zones of inhibition on Kirby-Bauer plates