JEFFERSON COLLEGE Jefferson Community and Technical College MLT 207: Introduction to Clinical Diagnostic Microbiology Module 9: Biochemical Identification for Gram-Negative Rods Study Sheet 1. What sugars are used in TSI media? 2. What is the most common sugar used in O/F basal media? 3. What tests are used to help measure carbohydrate utilization? 4. What tests are part of the IMVIC? 5. What tests measure amino acid utilization? 6. What are some principles the multitest systems can be based on?
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FOR LABORATORY EXERCISES, YOU WILL BE RESPONSIBLE FOR: 1. Knowing the media, procedure, and method of interpretation of results for the Kirby-Bauer (disc diffusion) antibiotic sensitivity test. 2. Knowing the cellular mechanisms of action for all antimicrobial agents/antibiotics used in Laboratory Exercise 31. 3. Knowing how to determine the relative effectiveness of disinfectant chemicals by examining filter discs on a bacterial lawn. What is a zone of inhibition? Knowing all appropriate definitions and vocabulary concerning agents for the physical control of microbial growth Exercise 32. 4. Knowing how fermentation works and how it is detected using liquid carbohydrate media. What is a Durham tube, what is detected by it, and how does the positive response appear? Exercise 36. 5. Being able to interpret the results of TSI Exercise 38. 6. Knowing all tests of the IMViC theory components, reagents used, products detected results, and how they appear media used for each test Exercise 38. 7. Being able to identify unknown G+ and G- bacteria. Exercise 54 and 55.
Supreeta N.
Lab 15: Gram-negative bacilli 1. What is the structure of Gram negative cellular envelope? 2. Name at least three genera belonging to Enterobacteriaceae family. 3. Characterize Enterobacteriaceae: a) What is their Gram reaction? b) What is their shape? c) What body part is inhabited by these bacteria? d) What is the transmission route for these bacteria? e) What type of metabolism do they have? f) What are their oxygen requirements? g) Are they motile? h) What kinds of infections are caused by these bacteria? 4. What does EMB stand for? 5. What is the purpose of EMB (what kind of medium is it)? a) What kind of organisms are growing on this medium? b) What indicator do we use? c) What inhibitor do we use? d) What is the main sugar in the EMB medium and why? e) What color indicates sugar fermentation? 6. What is the purpose of Tetrazolium salt medium? a) What color indicator do we use in Tetrazolium salt medium? b) How do we determine motility using Tetrazolium salt medium? c) What are some other ways to check for bacterial motility? 7. What final products of sugar fermentation could be determined in Phenol red medium? a) What is the pH indicator for acid? b) What color indicates positive result for fermentation? c) What color indicates use of peptone (protein) instead of sugar nutrients? d) What is the indicator for gas? 8. What is the function of oxidase? What color of the oxidase reagent indicates positive result? How could you interpret positive oxidase test? 9. What is the substrate of Urease enzyme? 10. Which bacterium is positive for Urease? 11. What color indicates positive result for Urease and why? 12. What does SIM stand for? a) What indicator do we use for sulfur? b) What color indicates sulfur reduction? c) What indicator do we use for Indole? d) What color indicates Indole presence? e) What indicates bacterial motility in SIM medium?
Adi S.
Columbia CNA Agar MacConkey's Agar Carbohydrate Tests IMVC Gram + Gram - Selected Biochemical Tests for Gram Positive Bacteria Name Shape Arrangement Pigment Catalase Salt Tolerant Hemolysis Bacitracin Sensitivity Optochin Sensitivity Growth on Bile Esculin Hydrolysis Bacillus subtilis rod Singles, chains none + - β NA NA - - Staphylococcus epidermidis coccus clusters none + + γ, α NA NA - - Streptococcus pneumoniae coccus chains none - NA α resistant sensitive - - Enterococcus durans coccus chains none - slightly γ, α, β resistant resistant + + Micrococcus luteus coccus clusters yellow + - NA NA NA NA NA Selected Biochemical Tests for Gram Negative Bacteria Name Shape Arrangement Glucose Lactose Sucrose Indole Methyl Red Voges-Proskauer Citrate Esculin Hydrolysis DNase Oxidase Neisseria sicca coccus pairs NA NA NA NA NA NA NA - NA + Escherichia coli rod singles acid, gas acid, gas - + + - - - slight - Salmonella enterica rod singles acid, gas - - - + - + - slight - Serratia marcescens rod singles acid, gas - acid - - + + - strong - Enterobacter aerogenes rod singles acid, gas acid, gas - - - + + + slight - Pseudomonas fluorescens rod singles - - - NA NA NA NA NA NA + NA – not appropriate test for the specific microbe
Keemin L.
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