Micro Test 1
Micro Test 1 Mbios 305
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This 45 page Bundle was uploaded by Nate Biancardi on Monday February 9, 2015. The Bundle belongs to Mbios 305 at Washington State University taught by Dr. Wang in Fall. Since its upload, it has received 195 views. For similar materials see Microbiology in Biomedical Sciences at Washington State University.
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Date Created: 02/09/15
Chapter 1 01142014 Go Over Appendix I Importance of Microbes Who cares 0 Alternative fuel industries Using lipid and fat content from algae 0 Genetic Engineering Agro bacteria allow genes of interest to be incorporated into plant genomes for a better function 0 Food Spoilage and Food Preservation Ramstein Shwarz glas Thursday 116 Lecture 2 Spontaneous Generation 0 Life from nothing Diseases were bad luck for people who were getting sick 0 Try to bleed out infected patients 2 Early Microscopes Robert Hooke 16351703 0 Before golden age of microbiology O O 0 Most famous for work with physics Hooke s Law Wanted to be able to see things we couldn t with the naked eye First microscope he developed was basically a backwards telescope Antoni Van Leeuwenhoek O O 00 Before golden age of microbiology Trader specialist of textiles Cloth furs Wanted to view textiles more closely lnvented rst handheld microscope Better than Hooke s microscope Better resolution Not a well known scientist Not taken seriously compared to Hooke Theory of Spontaneous Generation Francesco Redi O O O Disproving this theory Looked at food spoilage to food preservation Wanted to see if maggots grew on meat where did they come from Put meat in open jar and maggots grew on it after weeks Flies could land and lay eggs on meat 0 Saw that no food source no maggots 0 Container with meat in it and cheese clothe Cloth with pores in it No maggots grew on meat Flies were not able to reach meat 0 Start of scienti c method for biology Louis Pasteur 18221895 Golden Age of Microbiology 18501950 0 O O OO 0 John Tyndall Antibiotics mark the end of the golden age Father of Microbiology First to show Microbes were the rst to cause disease Swan Necked Flask He created growth medium which was something like chicken broth with added sugar Things started to grow on the medium when there was an open container S Curve on the bottom of the Swan Flask trapped microbes and dust because of gravity When he tipped the ask the growth medium started growing something This showed that spontaneous generation does not exist and there must be something we cannot see 0 18201893 Golden Age 0 Repeats Pasteur s experiments but got mixed results 0 Used differed infusions growth media Some remained sterile Others still spontaneously grew 0 Endospores Tough resistant outer coat Typically only gram positive microbes form these Protect microbe that is dormant Example n Bacillus Antracis white football 0 Causes antrax n Clostridium Perfringens brown with green outside 0 Causes gang green I If dry and nutrients are low then these two go dormant These endospores were in the hay that Tyndall used and when the sample was boiled the endospores activated and things began to grow this appeared to be spontaneous growth Boiled media three times and gured out that he didn t have spontaneous growth Medical Microbiology 0 Robert Koch 18431910 0 Golden Age 0 Father of Medical Microbiology 0 Worked with liquid media that Tyndall worked with 0 Pure Cultures Demonstrated diversity I Could observe different things Isolated single colonies n Identify causes and effect relationships of microbes and disease 0 Diseases Speci c isolate Homogeneous culture Single species cause single appearances of disease 0 Koch s Postulates Memorize Criteria for causing infectious disease Scienti c Approach leads to improved diagnosis and treatment Has limitations but still used 1 The microbe found in all cases of the disease but is absent from healthy individuals 2 The microbe is isolated from the diseased host and grown in pure culture 3 When the microbe is introduced into a healthy susceptible host the same disease occurs 4 The same strain of microbe is obtained from newly diseased host and harvest it to nd the exact same microbe in diseased host These postulates can fail because a host could appear to be perfectly ne but can still carry disease Clostridium Perfringens o Bacillus Anthracis Geogemma Barossii Strain 121 o Penicillium Notatum T4 Phage Neisseria Gonnorrheae 3 Microbial Domains Prokaryotes Unicellular Archaea Unicellular Bacteria Original Ancestor for all of these bridged into bacteria archaea and eukaryotes Constantly interacting Eukaryotes Unicellular o Algae o Fungi yeast 0 Protozoans Protists amoebas Multicellular o Algae o Helminthes Round worms Tapeworms Viruses Non living or abiotic Nucleic acid protein coat capsid 0 RNA or DNA Require host to multiply Eveloped 0 Host cell derived outer member 0 Use piece of host cell and wrap themselves in it o Occurs when exiting host cell NonEveloped 0 Naked 0 Outer coating is capsid Viroids Nucleic acid only o No capsid 0 Associated primarily with plant disease 0 Hosts are primarily plants 0 Always Single stranded RNA Pdons O Misfolded protein only Abnormal on rmation of already existing protein Neurological Diseases Mad cow disease in cows Spreads by misfolding other prions Observing the Microbial cell Microscopy 0 00000 SizeShape Resolution resolving power Allows us to categorize microbes Focus on what you would be able to see with that microscope Can you see two 2 red blood cells vs 1 red blood cell Human eye resolves 150 micrometers in diameter or 1 millimeters long Can notice a paramecium Procholorococcus marinus 4 nanometers Staphylococcus aureus 9 nanometers E Coli 25 nanometers Red blood Cell 8 nanometers Paramecium 100 nanometers OOOOO Instrumentation Light Microscopy 0 As small as 10quot6 meters 0 Bright eld typical 0 Electron Microscopy o 10quot8 meters 0 Scanning microscope 0 Transmission microscope Atomic Force Microscopy o 10quot9 meters 0 3D XRay Crstallography NMR Spectroscopy o 10quot10 meters or Angstrom Light BrightField and Staining Simple Stains 0 Add dark color or contrast 0 Methylene blue 0 Works by binding to negative charge of phosphorous on membrane 0 Bright Field What sample sits on 0 Differential Stains o Gram Stain Named after Hans Christian Gram 18531938 Wanted to see if he could differentiate human derived cells from microbial cells Worked on n Streptocaccus pneumoniae which stains purple n Klebsiella pneumoniae which stains red Saw that both these manifested pneumonia so it does not follow Koch s Postulates o Separates mixed microbes Must be able to tell one thing from another Gram Staining Procedure 0 Add methanol to x cells to surface and air dry 0 Gram Positive cells are xed to slide surface Bacillus Anthracis o Gram Negative cells are xed to slide surface E Coli Add crystal violet stain o Gram Positive Crystal Violet stains cells reversibly o Gram Negative Crystal Violet stains cells reversibly Add iodine which binds stain to gram positive cells 0 Gram Positive lodine Complex with crystal violet increase stain retention 0 Gram Negative Stain remains reversible o lodine Mordant Wash with Ethanol o Gram Positive No change 0 Gram Negative Stain is removed from gram negative cells Add Safranin Counterstain o Gram Positive Remain Dark Purple 0 Gram Negative Counterstains the cell usually pink Other Differential Stains Acid Fast Stain o Mycobacteria M Tuberculosis and M Leprae o Technically gram positive o Waxy cell wall inhibits Gram Staining o Carbolfuschin acid methylene blue counterstain Spore Stain o Malachite green detects Bacilli and Clostridia Gram Positive Morphology or Shapes Bacillus Rod Coccus Sphere Spirochetes Corkscrew Vibrio Curved Rods Electron Microscopy 10quot8 meters 0 Scanning or SEM and Transmission or TEM Used to re electrons and the bounce of electrons is what you see 0 Vacuum No live samples 0 Samples will be destroyed from this process 10quot9 Meter and Better Instrumentation Atomic Force Microscopy o 10quot9 meters 0 3D o Allows for live samples 0 XRay Crystallography and NMR Spectostrophy o 10quot10 meters Pdons o 1997 Nobel Prize Winner Stanley Prusiner o lnfective Protein Only Diseases 0 CreutzfeldtJakob CJD 0 Bovine Spongiform Encephalopathy BSE cows 0 Scrapie sheep 0 Chronic Wasting Disease deer and elk o Feline Spongiform Encephalopathy FSE cats 0 Maintain their folding formation and are extremely hard to destroy Play a structural Role Slipknot The Virus of Life January 23 Know Bacillus Anthracis and Clostridium Perfringens Gram Positive Chemohetero organitropes Mechanism 0 Normal PrPAC vs Abnormal PrPASC o C cellular 0 SC scrapie o Misfolded Protein misfolds other proteins 0 Aberrant conformation runs into normal conformation and misfolds it so it looks like itself 0 Build up more and more of the abnormal proteins and this ends up affecting nervous system 0 Aggregation Types 0 Familial Genetic component naturally occurring amino acids in a family that could favor misfolding o SporadicSpontaneous Occurred immediately could not recognize anyone essentially a vegetable 0 Acquired Can be acquired from a cow mad cow disease Chronic wasting disease can be spread by a decomposing deer s prions surviving in environment and another animal eats the prions and acquiring the disease as well Blood born contact issue In New Hampshire someone with prion disease was had surgery and utensils were not sterilized and it was given to another patient Viroids o Infective RNA 0 No Capsid Requires host RNA dependent RNA polymerase for replication Hepatitis D 0 Only human viroid known at this time 0 Associated with people with hepatitis B o No body has hepatitis D by itself 0 Potato Spindle Tuber Viroid o Viroid elongates the potatoes shape 0 Prevents the potato to convert its own metabolism 0 Destroys whole crops 0 Hard to get rid of 0 Not a lot of ways to treat this disease Viruses o Noncellular particle 10500 nm 0 Same size as our smallest bacterium 0 Could see with NMR microscope XRay crystallography and Atomic Force Microscope Must infect a host cell to reproduce Have parts of white blood cells 0 White blood cells are 810 micrometers o Allows them to consume viruses 0 Includes 0 Nucleic Acid DNA or RNA 0 Capsid Viruses and Hosts 0 Host Range 0 Single Species 0 Example HIV FlV feline canine parvovirus Usually only exists in one species 0 Broad Range 0 Example West Nile mosquitos birds us Exists in different species 0 MammalianHuman vs Plant 0 Tobacco Mosaic Virus TMV 0 Useful in terms of biotechnology Bacteriophage Phage o lnfects E Coli and or other bacterial species E Coli gram negative 0 T4 phage is drawn out on slide 31 0 Picture shows phage trying to get in purple and phage already in black dots How to Classify Viruses Physical Structure Genome Structure 0 RNA vs DNA 0 Single vs Double stranded Enveloped vs Naked 0 Outer lipid membrane derived from the host In uenza is example 0 No membrane on outside just proteins and nucleic acid T4 is example Baltimore Group 0 Double Stranded DNA 0 Uses its own or host DNA polymerase for replication Baltimore Group II 0 Single Stranded DNA 0 Requires DNA polymerase to generate a complementary strand Baltimore Group III 0 Double Stranded RNA 0 Requires RNA dependent RNA polymerase to make mRNA and genomic RNA 0 Baltimore Group IV 0 Single Stranded RNA 0 Requires RNA dependent RNA polymerase to make a template for mRNA and genome replication 0 Baltimore Group V Phage Life Cycle Classi cations o Lytic Virulent Cycle Replication and release host cell death 0 Lysogenic Temperate Cycle Integration of viral nucleic acids into host genome Host survives Stress or random events lytic cycle a Sign or trigger to switch to lytic cycle a Host dies Usually all of them move to lytic cycle 0 Slow Release Cycle Slower growing host cell survives Disadvantageous for survival of host from metabolic standpoint 0 Basics 0 Host recognition and attachment Hijacking cell surface receptors In Not meant to bind viruses but virus can match receptor and fool the host cell If host cell does not have speci c receptors then phage will not be able to get to host cell 0 Genome entry Only the genomic material enters and the other parts are left outside Only RNA or DNA enter 0 Virion Assembly Hijack ribosomes and other things to assemble more and more viruses 0 Exit and Transmission Ends up usually killing or exploding host cell Lytic Cycle 0 T4 0 Noneveloped Group or double stranded DNA 0 Virulent Stages 0 Express viral proteins 0 Digest host cell DNA 0 Synthesize capsid viral DNA and assemble o Lysis or cell bursting releasing virions Phage attaches to host cell and inserts DNA Linear double stranded DNA cyclizes to circular DNA Viral DNase cleaces host cell DNA Cell synthesizes capsid proteins Cell replicates phage DNA DNA is packaged into capsids Phage lyses cell and progeny phages are released Can happen in 60 minutes or less Lysogenic Cycle Lambda noneveloped and Baltimore Group I Temperate Stages 0 Form prophage Integration of virulent DNA to host cell genome o Reproduces with Host DNA 0 Excision l Lytic Cycle Survival inside vs outside Random event or stress Phage DNA integrates into host genome to form prophage Integrated phage DNA reproduces with host genome Integrated phage DNA replicates with host genome 0 Stress induces excision of phage DNA Phage recombines by rejoining the ends of its phosphodiester chain and enters the lytic cycle Slow Release 0 M13 Nonenveloped Group II 0 Cell Survives o Slows host cell growth Limited resources 0 No lysis Stages 0 Phage DNA replication and capsid synthesis 0 Assemble virions o Phage inserts single stranded DNA Phage DNA forms a double stranded circle 0 Cell replicates circular single stranded DNA Phages assemble and exit without lysis While phages reproduce and exit cell the cell reproduces slowly Less destructive and slower process What s in a Prokaryotic Cell 0 705 Ribosomes o 50 of antibiotics are 705 ribosome inhibitors 0 DNA quotnucleoidquot 0 Orange gure inside purple on slide 2 o No nucleus so this is where DNA and proteins are 0 Cellular Envelope 0 All prokaryotes have a cell envelope 0 Image shows gram negative microbe What is the Cell Envelope 0 The bag that holds everything else inside 0 Made of 0 At least 1 maybe 2 cell membranes 0 Gram negative have 2 cell membranes and thin cell wall 0 Cell Wall peptidoglycan Not cell envelope but part of it o Other structures Similarities and Differences between Gram Positive and Gram Negative bacteria Gram Positive 0 Very thick cell wall 0 One membrane Gram Negative 0 Thin cell wall 0 Two membranes What s the Same Between the Gram Positive and Gram Negative 0 Inner cell membrane made of lipids Cell wall made of peptidoglycan o No lipids Some proteins 0 Involved in transport of nutrients and other items Cell Membrane Similarities Always a semipermeable phospholipid biayer Tails of phospholipid biayer are hydrophobic Heads of phospholipid biayer are hydrophilic Sometimes contains other components that are not found in ALL bacteria Bacterial Membrane Lipids Structure 0 Dglycerol backbone CH20H3 0 Fatty acids esteri ed OCO with glycerol Saturated or unsaturated o Esters are susceptible to hydrolysis or breaking through water Bacteria are going to have a certain amount of saturated fatty acids and a certain amount of unsaturated fatty acids Unsaturated Cis fatty acids throw in kinks which allows membranes to be more exible o The increased exibility helps the microbe in cooler conditions Microbes living in higher temperatures usually have more saturated fatty acids to make it more rigid Different about Microbes Gram Stain Color 0 Gram Positive Purple 0 Gram Negative Red 0 Number of Membranes o Gram Positive 1 membrane 0 Gram Negative 2 membranes 0 Cell WallPeptidoglycan Thickness o Gram Positive Thick Outside of Cellular membrane 0 Gram Negative Thin Cell wall is in periplasm Teichoic acids 0 Gram Positive have that pass through the cell and bind to cellular membrane and provide additional support 0 Gram Negative do not have teichoic acids o Teichoic acids are part of cellular envelope 0 Porins o Gram Positive don t have porins o Gram Negative porins allow for transport only on the outer membrane Lipopolysaccharide 0 Only in Gram Negative 0 Sugar Coating o Gram Positive have sugar structures on outside Lipopolysaccaride LPS 0 Only in Gram Negative Endotoxin 0 Foreign or toxic o Initiates huge immune response 0 3 Parts 0 Lipid A endotoxin part recognized by immune system that shows it is bad Outermost layer of Gram negative cellular membrane Same in all microbes with LPS 0 Core polysaccharide Same in all microbes with LPS o Ospeci c polysaccharide or Oantigen Varies between microbes Can often be anywhere from 100200 sugars in length Hangs off into environment where microbe is living Allows us to identify microbes Cell Wall Peptidoglycan Peptide chains 0 Mini proteins 0 Sugars o No Lipids Not all bacterial have cell walls Gram Negative Peptidoglycan Same sugar glycan subunits Backbone is glucose memorize glucose 0 Use Slide 2 to memorize drawing 0 Nacetylglucosamine NAG on left of slide Structure of glucose with amine at carbon 2 o Nacetymuramic acid NAM in middle C3 lactate amide bond with peptides Nacetyl is just part at carbon 2 Beta14 linkage o Glycosidic bond two sugars connected together 0 Bond is an ether ROR o Goes between NAG and NAM o Lysosime destroys this bond Same locations for cross bridges 0 3rd and 4th amino acids of different peptide chains Different Peptidoglycan Peptides o Gram Negative Neissaria gonorrheae Bacillus Salmonella typhimurium Bacillus Treponema pallidum Spirochete Mostly conserved o LAlanineljDGlutamateljmDAPljDAlanine 3 always mesoDAP Different cross bridge structures between gram positive and gram negaUve ln GramNegative Microbes 0 Direct amide bond between two amino acids mDAPDAla of separate peptide chains Gram Negative Peptidoglycan o Memorize Sugars linked through Beta 14 glycosidic bonds at the top 0 0 Direct amide cross bridge between top mDAP and bottom DAla NAG and NAM connect to LAla on both top and bottom 0 Look at slide 0 No double cross bridge Gram Positive Peptidoglycan 3 Varies between species 0 Other positions are conserved o Bacillus Anthracis o LAlaljDGluljmDAPDDAla Staphylococcus aureus 0 LAlaljDGluleLysljDAla Cross Bridge Structures 0 Bacillus Anthracis has direct cross bridges o Staphylococcus aureus has interpeptide cross bridges Additional amino acids like Glycine Top LLys connects with Bottom DAla by 5Gly Specialized Structures PiliFimbriae O 0 Not found in every microbe Conjugation bacterial sex 0 Adherence stickiness o Motility use for transportation 0 Microbe needs a separate pili for each function 0 Flagella 0 Only found in Gram Negative bacteria 0 Used for motility transportation 0 Need energy to function 0 Found in Escherichia Coli Endospores 0 00000 O Resistant to heat drying UV and 02 Dormant awakens upon environmental cues Extension and stronger version of cell wall Encase themselves in a thick coat of peptidoglycan Survive up to decades Tyndall awakened the spores in his rst heating which made his medium cloudy when it was supposed to be sterile Clostridium Perfringens Obligate anerobe cannot be exposed to 02 If it has endospore then it can be exposed to 02 o Bacillus anthracis forms endospores as well Know Strain 121 Geogemma barossi for Archaea Coccus No Cell Wall Make a table that show differences between archaea and bacteria 0 Slide 11 and 12 Bacteria Cell Membrane o Backbone Dglycerol o Lipids unsaturated and saturated fatty acis Unsaturated have kinks that allow for more uidity survive colder temperatures Saturated are more rigid o BackboneLipid Bonds Esters Cell Wall 0 Sugars NAG and NAM o Glycosidic Bonds Beta 14 Archaea Have cell envelope 0 Prokaryotic Lives at really high temperatures Cell Membrane Stucture o Backbone Lglycerol o Lipids branchedcross linked saturated fatty acids Branches addition of methyl group CH2 n Thickens material and adds hydrocarbons Does not want unsaturated fatty acids Cross linked carboncarbon bonds between chains 0 BackboneLipid Bonds Ethers more stable n ROR Cell Wall 0 Sugars NAG and NAT U Know NAT structure 0 Glycosidic Bonds Beta 13 o PeptidesCross Bridges Lamino acids only Different cross bridges All L before amino acid Complex Media Rich or unde ned Each batch slightly different Ingredients like yeast extract peptone o No speci c chemical formula 0 Don t know exactly what is in there Media like LB TB terri c broth blood agar Still have numbers De ned Media Synthetic Exact amounts and formulas Minimal Media is an example 0 Known molecular formulas 0 See that each part of the media is DEFINED Contains salts Selective Media 0 Life or Death Natural Selection 0 Examples 0 Antibiotics resistant vs susceptible LB ampicillin To select for a resistant you give the bacteria a gene that kills ampicillin 0 High Salt osmotolerance 0 pH 0 Dyes Crystal violet is a selective agent for Gram Negative and against Gram Positive Differential Media 0 Differences in metabolic traits among strainsspecies Not necessarily life or death 0 Generally associated with enzymatic activity 0 Blood agar o Streptococcus and Staphylococcus Gram Positive Have hemolysin which destroys red blood cells Can differentiate these two between other microbes The hemolysin allows a clearing or greenish color in the blood agar OOOO o Occurs from oxidation of iron Mostly survival of microbe Selective and Differential Media MacConkey agar o The Pancreatic parts at the top shows that it is complex because there is no de ned chemical formula 0 Crystal violet shows that it is differential along with bile salts Selects for Gram negative and Gram Positive die 0 Dye crystal violet inhibits Gram Positive 0 Bole Salts inhibit most Gram Positive Differentiation o Lactose pH indicator Fermentation pathways Red Colonies Fermenters Escherichia coli Colorless non fermenters Salmonella a Does not spew out acid so will not change color Treponema pallidum cannot survive Macconkey agar because it is not able to be plated Nutritional Requirements Troph nutrient requirements Classify Based on 0 Energy Source Phototrophs vs Chemotrophs 0 Carbon Source Autotroph vs Heterotrophs 0 Electron Source Lithotrophs vs Organotrophs o Combining Attributes Photoautolothotrophs Energy Source Phototrophs 0 Light 0 Anabaena spiroides anol Prochlorcoccus marinus Chemotrophs 0 Chemical compounds 0 Salmonella typhimurium Carbon Source Autotrophs 0 Fix C02 into organic compounds 0 Anabaena spiroides anol Prochlorococcus marinus Heterotrophs o Consume organic compounds make biomass and wastes C02 0 Salmonella typhimurium and Bacillus anthracis Electron Source Metabolic Redox 0 Reduction oxidation chemistry 0 Oxidize electron donors Lithotrophs 0 Electron donor oxidize inorganics like water 0 Anabaena spiroides takes electrons off of water for photosynthesis Organotrophs 0 Electron donor 0 Oxidize organics like food 0 Salmonella typhimurium and Bacillus anthracis Anabaena spiroides photoautolithotroph Strain 121 chemoautolithotroph Prokaryotic Growth Binary ssion vegetative growth Generation or doubling time Nt NO 2quotn 0 Mt number at time t 0 N0 number at time 0 or start time o n number of doublings Examples Ideal Conditions 0 Salmonella typhimurium and Escherichia coli 30 minutes 0 Neisseria gonorrheae 60 minutes 0 Treponema pallidum 33 hours Detecting Growth Grow in liquid media and measure turbidity o Spectrophotometer AbsorbanceOD optical density 1 OD600nm Reading 10quot8 colony forming units or cells per mL of culture or CLUmL Not the best tool to use to detect infectious microbes Need to be able to take OD reading and put it into doubling equation BillyJoel I Go to Extremes Phases of Prokaryotic Growth 0 Associated with a closed system like a culture 0 Not an open system like bloodstream Look at Graph on Slides Lag Phase 0 Whenever you start the culture 0 Time will be different with different microbes Bacteria and archaea grow at different rates 0 Cells are trying to get to binary sion and are in a dormant state 0 Early Log exponential phase 0 Binary Fision dominates growth curve 0 Many Primary metabolites are produced Vitamins are made and are cofactors for enzymatic reactions Wastes Form and begin to poison the microbes because of closed system 0 Very few bacteria present 0 Tons of nutrients present 0 No wastes present C02 0 Late Log Phae 0 Lots of doubling occurring 0 Not as much nutrients available 0 Waste is building up o Endospores start to form from stress Bacillus anthracis and Clostridium perfringens Good thing for bacteria Bad for bacteria 0 Secondary metabolites form Quorum signals form and are small chemical compounds that microbes use to communicate Anitbiotics form to kill other microbes Stationary Phase 0 Rate of Death Rate of Doubling o Wastes are really taking effect 0 Survivors are able to produce Endospores antibiotics and quorum signals 0 Death Phase 0 Small amount of grow 0 Mostly a microbes die Way too many wastes Large lack of nutrients Normal vs Extremophiles There are no archaea that are known to be pathogenic Temperature 0 Normal 0 Mesophiles any pathogenic microbe we have talked about They live in food or humans Optimal temperature is 1545 degrees C Most have optimal growth at 3740 degrees C Extremophiles o Psychrophiles Around 15 degrees C or lower is optimum a Close to freezing Fluid or unsaturated membranes 0 Thermophiles 5570 degrees C is optimum Both archaea and microbes can live here 0 Hyperthermophiles 80 degrees C is optimum Crosslinked saturated membranes In Ether bonds Strain 121 Geogemma barossi n Optimum close to 110 degrees C Proteins protect DNA at high temperatures Salt Osmotic Stress l plasmolysis membrane and cell wall separate 0 Too much salt causes death of cell 0 Proteins and teichoic acids start to come apart Normal 0 25 NaCl Includes humans and lakes and rivers Escherichia coli 0 Osmotolerance Staphylococcus aureus Ok at concentration up to 10 NaCl n Located on skin and want to survive sweat Extremophiles o Halophiles Require 3 NaCl Oceans 0 Extreme halophiles Normal Require 1020 NaCl Great Salt Lake Microbes have evolved to require these concentrations of salt pH o Neutralophiles pH 58 or optimum is around 7 Almost every microbe we have talked about Bacillus Anthracis Extremophiles o Alkalophiles pH 9 0 Acidophiles pH less than 5 Lactobacillus acidophilus a Run a lot of fermentation I Found in yogurt n Gram Positive a Make environment acidic to kill off other microbes Use of vs Response to Oxygen o NONE OF THESE CONDITIONS ARE EXTREME Two distinct classi cations 0 Use of Energy production with or without 02 Use oxygen n Aerobic Respiration Don t use oxygen In Anaerobic respiration n Fermentation 0 Response to Ability to detoxify reactive oxygen species Strict aerobes Facultative anaerobes microaerophiles aerotoerant anaerobes obligate anaerobes Use of Oxygen Obtain energy from light or chemical compounds 0 Phototroph or chemotroph Removed Electrons from chemical compounds and transfer them to terminal electron acceptor which leaves the cell Waste product 0 Lithotroph or Organotroph o If oxygen is terminal electron receptor aerobic respiration 0 Terminal electron receptor Nitrate Sulfate Sulfur anaerobic respiration Inorganic compound 0 Terminal electron receptor organic compound fermentation Respiration 0 Use electron transport system to make proton motive force 0 Proton Motive Force is used by synthase to make ATP Oxidative Phosphorylation Fermentation o No electron transport system o No advantageous because no ATP Response to Oxygen O2 l reactive oxygen species ROS Toxic Byproducts oxidative stress 0 Superoxide radical free electron on oxygen Free electron wants to form covalent bond Reacts with hydrogen peroxide 0 Hydrogen Peroxide H202 o HOdot hydroxyl Radical that reacts fast Avoid Hydroxyl Radicals Detoxi cation o SOD superoxide dismutase Enzyme that generates peroxide but consumes superoxide radical 2 O2 superoxide 2H l 02 H202 o Catalase and Peroxidase Consume Peroxide 2 H202 l 2H20 02 Strict Obligate Aerobe 0 Has SOD and Catalase and Peroxidase 0 Only respires aerobically Requires OZ 0 Cant respire anaerobically or ferment 0 Examples Gram Positive Mycobacterium tuberculosis Gram Negative Neisseria gonorrheaa Facultative Anaerobe 0 Have SOD and Catalase and Peroxidase o Prefers to respire aerobically Grows better with OZ 0 Can respire aerobically or ferment 0 Examples Gram Positive Bacillus anthracis Gram Negative Escherichia coli Aerotolerant Anaerobes 0 Have SOD and Catalase and Peroxidase Cannot respire Can only ferment Obligate fermenters Doesn t care about OZ Examples Gram Positive Lactobacillus acidophilus Microaerophile 0 Little SOD and Catalase and Peroxidase o Respire aerobically o Requires low levels of OZ Higher 02 levels toxic 0 Examples Treponema pallidum Strict obligate anaerobe o No SOD Catalase or Peroxidase o 02 is toxic o Respires anaerobically and or ferment 0 Example Gram Positive Clostridium perfringens OOOO
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