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Exam 1 study guide

by: Katy Cook

Exam 1 study guide Bio 439

Marketplace > Purdue University > Bio 439 > Exam 1 study guide
Katy Cook
GPA 3.45
Biology 439- Microbiology
Dr. Walter

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Biology 439- Microbiology
Dr. Walter
Study Guide
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This 12 page Study Guide was uploaded by Katy Cook on Tuesday February 10, 2015. The Study Guide belongs to Bio 439 at Purdue University taught by Dr. Walter in Fall2014. Since its upload, it has received 640 views.


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Date Created: 02/10/15
Organisms The 10 we looked at in the beginning Lab 1 Saccharomyces cerevisiae Single cocci ovoid budding BCG agar green with white surrounding pH indicator odor round slimy domed yeast smell Corynebacterium diphtheriae Clubbed cells Vshaped arrangement G positive black on K2Te03 agar domed tiny anaerobic respiration Streptomyces Filamentous G positive cluster crusty odor orangered diffusion clumpy in broth domed native musty Bacillus thuriengiensis subtilis cereus Rods G positive bright spores chains protein crystals large flat native spreading margin fuzzy a I i332 o 9quot g b0735 0 5 Cquot lo l h V E 0 g claw 5 i F 0 l o f g a l F mm is U 9115quot Bias 3 H 0 if m 6 9 go I lo 939 7 V u 3 Earths o a a C be 0 Q g 0 on 393 393 i g 39 i Staphylococcus Cocci clusters G positive rings of clearing on blood agar native domed Streptococcus Enterococcus faecais closely related Cocci chains G positive lysis of blood around small colonies domed partial lysis around S pneumoniae fastidious KlebsieIa pneumoniae Single rods G negative capsule around cell glycocalyx native slimy surface white domed Escherichia coli Single rods G negative on EMB native on TSA Proteus mirabilis Single cocci G negative pink colorless on EMB swarming motility on TSA native no swarming at cooler temp domed Serratia marcescens Single rods G negative redorange on TSA green metallic on EMB domed Pseudomonas aeruginosa Single rods G negative flat green on TSA fruity odor more pigment in older culture uses ED glycolysis robust Calculations Titer plating and dilutions dilution factors serial dilutions Titer cc X TDF X PF cfuml PF 1vol plated Poisson distribution the s SORT m part but also the Prm mAr rl eAm part The variance 8A2mean S SQRT mean 68 of values between 18 95 of values between 28 Prm mAr equotm r maverage number of p outcomes in infinite number of trials rnumber of trials that results in the p outcome nnumber of statistical trials PQ probabilities of 2 outcomes Media prep problems quotbucketquot problems for dilution fold concentrate 1 OX means what weightvolume molarity and also how to sterilize things Stock Diluent Final Product C V Foldconcentration l diluted 10X Weightvolume 1g1OO ml Molarity molL Not autoclave Heat liable antibiotics amino acids aromatic will all denature Not together sugars with phosphate salts Incompatible ions precipitate l group II w SO4PO4 Sterile media Filter sterilize CAA Prepare medium stopper flask and autoclave Cool add glucose add salts add sterilized CAA solution Nutrient agar weigh appropriate amount of nutrient broth suspend in DI water add agar add foil and autoclave Pour aseptically into two petri dishes cool and solidify Growth problems applying the growth rate equation as for homework 2 exercise 10 exercise 8 Specific growth rate constant u measured in units of 1time Generation timetime required for number of cells to double u nN nNo tftO NN0equotut n2u td n2tdu Growth yield what Yield represents about cell metabolism in addition to calculations like those in Exercise 9 and 10 Yield amount of product cells mass of cells Amount of limiting reagent mg Glucose mol Glucose Y Concentration of carbon in cell culture Concentration of Glucose The yield is equal to the biochemical efficiency Y ODfODi x 045 mg cell Cml Glucose x10 mgml Techniques Sterile transfer pipets inoculating loops tubes tube caps foam plugs flasks etc Making slides fixed mount versus wet mount Wet mount drop of a suspension of microorganism under a coverslip Microscopy focusing using oil immersion measuring things under the microscope Calculate size in recticle units ru and convert to um Each line on stage micrometer are 10 um wide Stains Simple stain Gram stain Negative stain Simple stain ex methylene blue Gram stain Crystal violet 20 sec Rinse lodine 60 sec Ethanol Rinse Safranin Rinse Negative stain Small drop of India ink Suspend in the ink small portion from colony Spread to size of nickel Make a smear with cover slip Air dry and heat fix Counterstain for 30 sec Rinse and blot dry Plating Streak plate Spread plate Pour plate Using the Spec20 to measure Optical Density zeroing and blanking the Spec taking readings Background material Resident vs transient flora quotglove juicequot extraction with lecithin effectiveness or lack thereof of washing hands various ways Ethanol was not very effective That plate still contained some of the transient flora that was not killed The dish where hands were washed from 30 seconds had only one small colony so it was either transferred incorrectly or something else caused them to be killed The last dish from washing hands for 2 minutes was contaminated with serratia Parts of a microscope and what they are used for Sewage treatment process primary secondary and tertiary stages flocs activated sludge what organisms you would see Primary designed to remove suspended and floating solids Mechanical treatment Secondary Removes dissolved organic matter by microbes consuming the organic matter as food and converting it to energy Followed by additional settling tanks to remove suspended solids Activated sludge Tertiary Remove additional phosphorous and nitrogen Disinfection with chlorine Activated sludge process for treating sewage using air and biological floc composed of bacteria and protozoa Biological floc reduces organic content of the sewage composed of bacteria and protozoan flora sometimes rotifers 1 Unstained raw sewage few small scattered organisms hard to see 2 Unstained activated sludge some organisms but transparent 3 Stained raw sewage Floc with organisms around it spirilla 4 Stained activated sludge Many organisms and filaments Gram positive versus Gram negative cell walls what they look like why the Gram stain differentiates them etc Some bacteria are decolorized after being dyed with crystal violet if flooded with alcohol for a specific period of time Other bacteria Gram positive were decolorized at a much slower rate Gram positiveness is characteristic of bacteria sensitive to penicillin an antibiotic which inhibits the synthesis of the thick peptidoglycan cell wall Gram reactions seem to be a mostly physical phenomenon the dehydrating activity of the alcohol closes pores in the thick peptidoglycan layer of the cell wall of the Gram positive organisms The alcohol therefore is only able to leach the crystal violet at a slow rate Alcohol readily permeates the lipoprotein and thin peptidoglycan of Gram negative cells so it can leach the crystal violet at a relatively high rate Definitions about growth and media various quottrophsquot robust vs fastidious differential selective complex defined growth factor enriched minimal nitrogen carbon electron energy sources organic vs inorganic oxidized vs reduced some review from your Chem classes here too Fastidious Complex nutritional requirement Needs specific nutrients Robust Only requires macromolecules N P and S Selective selectively allow the growth of some organisms while inhibiting growth of others Differential Allow to distinguish between two organisms Complex More than minimal nutrient needs not all of ingredients characterized Defined Provide basic minimal requirements for growth Growth factor amino acids nucleotides vitamins fatty acids Enriched more than minimal nutrients Minimal simplest nutrients to survive Carbon Source Organic CH bonds l heterotroph Inorganic C02 autotroph Energy source light l phototroph Redox reactions I chemotrophs Electron Source Organic CH4 C6H1206 I organotroph Inorganic Sulfide ammonium lithotroph Growth phases and what happens in each phase Lag phase time of adjustment or adaptation little or no increase in cell number or CD Cells restructuring their complement of enzymes in order to grow exponentially Log phase All of the cells divide in time required to double their mass Cell mass increases exponentially Stationary component of medium becomes limiting build up of inhibitory end product run out of 02 get too crowded still divide but one cell dies so no change no increase in OD Death lysis more than division decline Persister cell highly antibiotic resistant not mutants activate stress responsive genes 1 of population low metabolic rate don t divide growth versus survival metabolism Exponential versus stationary phase Concept of growth yield what it means carbon vs energy yield different pathway efficiencies EM vs ED glycolysis TCA cycle with or without glyoxylate shunt aerobic vs anaerobic etc efficiency of various carbon and energy sources exercise 10 Growth yield biochemical efficiency conc Carbon conc Glucose More Glucose More Carbon more energy yield higher OD more cell concentration ED pathway is less efficient produces less ATP Phosphorylation occurs before oxidation then hydrolysis Reaction yields only 1 ATP per glucose molecule Oxidation before cleavage Glyoxylate cycle anabolic variation of TCA cycle Allows cells to utilize simple carbon compounds as a carbon source when glucose not available Bypasses the decarboxylation steps that take place in the TCA cycle allowing simple carbon compounds to be used in the later synthesis of macromolecules Uses glyosylate and acetylCoA to produce malate Lysine is the least efficient acetate not very efficient Others about the same 05 Efficiency increases with increase in concentration of Carbon source Glucose lactose and aspartate highest efficiency HW problems Quiz questions


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