Idea Transcript
351 Practical I Review * This is just possible topics for exam, please review entire lab manual to study. This is not an inclusive list. Lab 1:*Also know safety information in the first 5 pages of lab manual.
Ex. 1-1 Nutrient Broth & Nutrient Agar Preparation *Please know how to prepare regular media, both broth and agar. *What is the difference between broth and agar? *Know the tools and protocols to making media. *What is an autoclave? Why do we autoclave? Ex.1-2 Common Aseptic Transfers and Inoculation Methods *Aseptic Technique: know special rules of aseptic technique described in lab manual. *Be able to show aseptic technique to TA *Know different processes of inoculation (broth to agar, agar to agar, etc) *Know different inoculation tools and what is use for each. Ex.1-3 Streak Plate Methods of Isolation *Be able to streak a plate properly in front of TA!! *What is CFU's? *Why do you use streak plate method?
NO DEMOS *Read manual
NO DEMOS *Read manual
Escherichia coli Staphylococcus aureus Micrococcus luteus Staphylococcus epidermidis Enterobacter aerogenes Serratia marcescens Bacillus subtilis
Ex.1-4 Spread Plate Method of Isolation *What is the difference between streak and spread? *How did you use the tubes as a dilution factor? *What would be the differences between a streak and a spread plate colonies? Lab 2: Selective Media *Know differences between selective, differential media, defined and undefined. *Know characteristic differences between Gram + and Gram - bacteria. Escherichia coli Ex. 4-1 Mannitol Salt Agar Enterobacter aerogenes *What makes this plate differential and selective? Staphyloccocus epidermids *What are the ingredients important to this media? Staphyloccocus aureus *What is the mannitol salt fermentation pathway? Alcaligenes faecalis *Why do (should) only Gram + grow on this media? Sporasarcina urea *Be able to identify demo bacteria that grow on this media and/or ferment mannitol. Escherichia coli Ex. 4-2 Phenylethyl Alcohol Agar Enterobacter aerogenes *What are the ingredients important to this media? Staphyloccocus epidermids *Why do only Gram + grow on this media? Staphyloccocus aureus *How does a positive result look compared to a negative growth that may occur? Alcaligenes faecalis *Be able to identify demo bacteria that grow on this media. Ex. 4-4 Endo Agar *What are the ingredients important to this media? *Why do only Gram - grow on this media? *What is the difference of coliform and non-coliform? *Be able to identify demo bacteria that grow on this media and/or ferment lactose. Ex. 4-5 Eosin Methylene Blue *What are the ingredients important to this media? *Why do only Gram - grow on this media? *Be able to identify demo bacteria that grow on this media and/or ferment lactose and/or little or more acid production.
Sporasarcina urea Escherichia coli Enterobacter aerogenes Staphyloccocus epidermids Staphyloccocus aureus Alcaligenes faecalis Sporasarcina urea Escherichia coli Enterobacter aerogenes Staphyloccocus epidermids Staphyloccocus aureus Alcaligenes faecalis Sporasarcina urea
351 Practical I Review * This is just possible topics for exam, please review entire lab manual to study. This is not an inclusive list.
Lab 2 Continued: Ex. 4-6 Hektoen Enteric Agar *What are the ingredients important to this media? *Why do only Gram - grow on this media? *Be able to identify bacteria within manual that will show hydrogen sulfide production. *Be able to identify demo bacteria that grow on this media and/or ferment lactose. Ex. 4-7 MacConkey Agar *What are the ingredients important to this media? *Why do only Gram - grow on this media? *Be able to identify demo bacteria that grow on this media and/or ferment lactose.
Ex. 4-8 Xylose Lysine Desoxycholate *What are the ingredients important to this media? *Why do only Gram - grow on this media? *Be able to identify bacteria within manual that will show hydrogen sulfide production. *Be able to identify demo bacteria that grow on this media and/or ferment xylose.
Escherichia coli Enterobacter aerogenes Staphyloccocus epidermids Staphyloccocus aureus Alcaligenes faecalis Sporasarcina urea Escherichia coli Enterobacter aerogenes Staphyloccocus epidermids Staphyloccocus aureus Alcaligenes faecalis Sporasarcina urea Escherichia coli Enterobacter aerogenes Staphyloccocus epidermids Staphyloccocus aureus Alcaligenes faecalis Sporasarcina urea
Lab 3: NSM (Nutrient Sporolating Media) *Why did you use this media? What will the stressed bacteria (if they can) form? *What is the importance endospores? Ex. 2-2 Colony Morphology *Know 5 basic categories to identify colony morphology. *Be able to identify specific bacteria by just their morphology. *Know the examples given within the lab manual as well. Lab 4: Ex. 3-1 Microscopy - light/phase *Know all of the parts of the microscope. *Know how the microscope works in order to project images. *Know terminology of microscopes (limit of resolution, numerical aperture, etc) *How do you find total magnification? *What is the difference between light and phase contrast microscopy? Ex. 3-3 Examination of Microbes *What are the three domains of life? Identify each demo bacteria into their domains. *Identify each demo bacteria into their classification according to manual (Phylum, sub-phylum, etc)
*What is the differences between prokaryotes and eukaryotes within regards to structure? *Be able to identify within the microscope each demo bacteria mentioned in manual. *Be able to identify the characteristics of each demo bacteria. *How do you perform a wet mount on an organism?
NO DEMOS. Read Manual! See above for demos
See Index Cards and Microscope slides
351 Practical I Review * This is just possible topics for exam, please review entire lab manual to study. This is not an inclusive list.
Lab 4 Continued: Bacterial Structure and Staining *Know arrangements of bacteria, differences between plural and singular nomenclature. *Know special arrangements of bacteria (tetrads, diplo-, staphylo-) *Be able to identify demo bacteria within a microscope by just their arrangement. Gram Stain Stations setup at your Ex. 3-6 Gram Stain lab bench. Utilize the demo plates *Know different steps to Gram Stain & what each step will do to the cell. (or tubes) in order to perform your *Know characteristic differences between Gram + and Gram - bacteria. own gram stain. During exam, you *Be able to perform a gram stain on an unknown to get shape, arrangement, and Gram results. will need to perform your own Gram *What is the difference between differential stains and simple stains? stain (without help of TA) to figure * Be able to identify demo bacteria according to Gram Results, shape and arrangement. out Gram results & shape. Ex. 3-9 Endospore Stain View some demo slides: *What is an endospore? What are the different type of spores that could be present? Bacillus subtilis *Why do you need to add heat? *Know different steps and timing to perform an endopore stain. *Be able to identify demo bacteria within the microscope of bacterium with endospores. Ex. 3-11 Flagella Stain Spirillum volutans - amphitrichous *Know the theory of a flagellar stain. Proteus vulgaris - peritrichous *Know the different types of flagella within bacteria. *Be able to identify demo bacteria with amphitrichous & peritrichous flagella. Lab 5: Ex. 2-1 Ubiquity of Microorganisms NO DEMOS. *Know the terms in bold. Read Manual! *Why was this experiment performed? What were your results?
Ex. 2-3 Growth Patterns of Slants *Know the bold terms identifying characteristics of slants. *Why do we use slants? *Know the characteristics of demo bacteria to be able to identify each organism by their slant. Ex. 2-4 Growth Patterns in Broth *Know the bold terms identifying characteristics of broths. *Why do we use broths? *Know the characteristics of demo bacteria to be able to identify each organism by their broth. Ex. 3-7 Acid-Fast Stain *Why would you need to do an acid fast stain? *What is the theory and development of the acid fast stain? *Be able to identify demo bacteria within the microscope of an acid-fact bacterium. Ex. 3-8 Capsule Stain *What are capsules? *How is a negative stain different from a regular stain? *What is the theory and development of the capsule stain? *Understand the relationship between virulence and capsule production, and medical significance of this.
*Be able to identify demo bacteria within the microscope of bacterium with capsules.
Micrococcus luteus Bacillus subtilis Staphylococcus epidermidis Serratia marcescens Escherichia coli Micrococcus luteus Bacillus subtilis Staphylococcus epidermidis Serratia marcescens Escherichia coli
Mycobacterium tuberculosis
Klebsiella pneumonia
351 Practical I Review * This is just possible topics for exam, please review entire lab manual to study. This is not an inclusive list.
Lab 6: Ex. 2-6 Agar Deep Stabs *Know aerotolerance terms. *Why do we use a stab? What is the difference between a deep and slant? *Be able to identify demo bacteria and their aerotolerance by viewing a deep. Ex. 2-7 Agar Shakes *How do you prepare an agar shake? *Be able to identify demo bacteria and their aerotolerance by viewing a shake. Ex. 2-8 Fluid Thioglycollate Medium *What is in thioglycollate to make the medium work to show aerotolerance? *What is in the oxidized portion (resazurin)? *Be able to identify demo bacteria and their aerotolerance by viewing a thioglycollate medium. Ex. 2-9 Anaerobic Jar *How does the gaspak system work? * What is in the indicator strip to show the absence of oxygen? *Be able to identify demo bacteria and their aerotolerance by viewing a aerobic/anaerobic plate Ex. 2-10 The Effect of Temperature on Microbial Growth *Know the bold terms identifying characteristics of temperature in microorganisms. *Be able to identify demo bacteria by temperature. Ex. 2-11 The Effect of pH on Bacterial Growth *Know the bold terms identifying characteristics of pH in microorganisms. *Be able to identify demo bacteria by pH. Ex. 2-12 The Effect of Osmotic Pressure on Microbial Growth *Know the bold terms identifying characteristics of osmolarity in microorganisms. *Know the difference between Hyposmotic, Isosmotic, and Hyperosmotic. *Be able to identify demo bacteria by osmolarity. Ex. 5-5 Catalase Test *Know the mechanism of catalase positive organisms and the enzymes associated to them. *What reagent do we use to see a positive result? *Be able to identify demo bacteria that are catalase positive. Ex. 5-6 Oxidase Test *Know the mechanism of oxidase positive organisms and the enzymes associated to them. *How dis you test for oxidase? *Be able to identify demo bacteria that are oxidase positive. Extra Activity Dilution Plating - Refer to 6-1 *Be able to figure out CFU, OCD, DF given a sample (see attached page) *Why do we use a dilution series for estimating bacterial counts. Extra Activity Growth Cycle of Escherichia coli *Be able to calculate generation time given a log graph. *Be able to properly label a log graph given time. *Know the different phases and what they mean in the life of the bacteria.
Escherichia coli Clostridium sporogenes Micrococcus luteus
NO DEMOS. Read Manual! This media does not sit well, gravity forces bacteria down. Ask TA about results.
Escherichia coli Clostridium sporogenes Micrococcus luteus Escherichia coli Clostridium sporogenes Micrococcus luteus Bacillus stearothermophilus Serratia marcescens Escherichia coli
NO DEMOS. Read Manual! NO DEMOS. Read Manual!
NO DEMOS. Read Manual!
NO DEMOS. See Attached Handouts