Exam 1 PowerPoints and Study Guide
Exam 1 PowerPoints and Study Guide 81382 - MICR 3050 - 001
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Chapter 1 The Evolution of Microorganisms and Microbiology What are microorganisms 39 organisms and acellular entities too small to be clearly seen by the unaided eye generally S 1 mm in diameter often unicelluar Exceptions Molds can see Without microscope and giant amoeba 1 in Worms too small to see with eye is not Copyright The McGraw Hill Companies Inc Permission required for reproduction or display Organisms and biological entities studied by microbiologists can be includes includes composed of composed of composed of composed of Yeasts Algae Escherichia Methanogens Molds Protozoa coli Slime molds The science of Microbiology Basic aspects Concerned with understanding individual groups of microbes Microbial physiology genetics molecular biology and taxonomy Understanding microorganisms has improved the understanding of other organisms The Importance of Microorganisms most populous and diverse group of organisms Estimate 510A30 bacterial on Earth found everywhere on the planet 9294 underground play a major role in recycling essential elements eX Nitrogen carbon sulfur source of nutrients some carry out photosynthesis Most beneficial or benign some detrimental in uence all other living things excellent tools for study Types of Microbial Cells 39 Prokaryotic cells lack a simple true membranebound nucleus this is not absolute some exceptions Eukaryotic cells have a membrane enclosed nucleus and other membranebound gganelles are more complex morphologically and are usually larger than prokaryotic cells Prokaryotic coll Cytoplasm Nucleoid Ribosomes Plasmid l l 05 pm Cytoplasmic Cell wall membrane Figure 21 a Brock Biology of Microorganisms 11e 2006 Pearson Prentice Hall Inc Figure 12b Brock Biology of Microorganisms 11e 2006 Pearson Prentice Hall Inc Prokayotic Herbert Voelz Eukaryotic cell Cytoplasmic membrane Endoplasmic reticulum Ribosomes Nucleus Nucleolus Nuclear membrane Golgi Cytoplasm Mitochondrion Chloroplast I Figure 21b Brock Biology of Microorganisms 11le 2006 Pearson Prentice Hall Inc 1O Acellular Infectious Agents not cells not true microorganisms Viruses consist of a protein coat and nucleic acid DNA or RNA some have additional layers require host cells to replicate cause a range of diseases some cancers Viroids and satellites infectious agents composed of m prions infectious proteins Examples mad cow disease 11 Eukaryotic cell 1000 nm 1 pm Figure 23c Brock Biology of Microorganisms 11e 2006 Pearson Prentice Hall Inc Prokaryotic cell Classification Universal Phylogenetic Tree three domain system based on a comparison of the DNA encoding small subunit ribosomal RNA 39 divides microorganisms into 3 Domains Bacteria Archaea the Ancient ones Eukarya includes all eukaryotes 1W DN sequencing C lls Gene encoding A G c 139 ribosomal RNA c nc s Figure 26 part 1 Brock Biology of Microorganisms 11e 2006 Pearson Prentice Hall Inc 13 sequencing AG 139 Aligned rRNA genesequences AGTCGCTAG 1 Sgequ nm AG c c G r 1 AG 3 a n a his i 5 f5 hyl 0 en etic t ree Figure 26 part 2 Brock Biology of Microorganisms 11e 2006 Pearson Prentice Hall Inc 14 Tree of Life ARC IIAEA Methan o g e ns 39EXtr emehalophiles Hyperthermophlles 4 39 1Tz i3 fi i 912 a u f 39 I EUKARYA quotTE iii is 5 rirzalaa fij aryouc ll 1 E Crown species Slime molds Flagellates Giardia Root of the tree EUKAw 0 39 L Mirquot r h 391 h r39J r A l V 391291rt4 Hr 4 m Figure 27 Brock Biology of Microorganisms 1 We 19 2006 Pearson Prentice Hall Inc Copyright The McGraw Hill Companies Inc Permission required for reproduction or display Bacteria gt g Q 93 D Q U K x 8 a 2 53 393 3 8 lt5 09 b G pg 69 95 3986 8 9 O OQ 63 e a 339 395 020 o 06 26 gm lt 0 0 df 3 Stramenopiles Crenarchaeota Alveoates Archaea Euryarchaeota l l rRNA sequence change Unresolved branching order 16 17 Domain Bacteria prokaryotes usually singlecelled majority have cell wall with peptidoglycan abundant in soil water air and inon other organisms some live in extreme environments Artic ice or acidic soils Domain Archaea prokaryotes distinguished from Bacteria by unique rRNA sequences lack peptidoglycan in cell walls have unique membrane lipids some have unusual metabolic characteristics many live in extreme environments The Microorganisms of DO m a i h EUkaI includes microorganism plants and animals eukaryotes Protists generally larger than members of Bacteria and Archaea algae unicellular photosynthesis Protozoa animallike unicellular moblie slime molds water molds funi yeast 19 mold 20 Origins of Life Microbial Evolution microbial fossils Swartkoppie and Archaean Apex chert 35 billion years old fossil record sparse Indirect evidence and scientific method study of extant organisms Copyright the McGrawHill Companies Inc Permission required for reproduction or display Millions E 5 56 of Years 8 m 2 ago g 0 o 7 m a Hominids first a ear 65 J 2 Cretaceous 144 g g o Jurassic 206 0 8 248 2 TrIaSSIc 225 mya Dinosaurs and mammals first appear 1 290 g Permian 354 E Q I Carboniferous 300 mya Reptiles rSt appear k a 417 l Devonian 443 2 m 490 o Ordovician 450 mya Large terrestrial colonization by plants and animals 543 g cambriaquot L520 mya First vertebrates first land plants 533 525 mya Cambrian explosion creates diverse animal life 900 U 5 8 600 g 1 5 bya Multicellular eukaryotic organisms first appear 1 8 a Z 1 a 2500 E g 25 20 bya Eukaryotic cells first appear amp O 3000 2 d M 4 I U I lt 3400 lt 35 bya Fossils of primitive filamentous microbes 3800 38 35 bya First cells appear 4550 How DidDo Microbes Evolve Bacteria and Archaea mutation Of genetic are haploid material increase genetic diversity w by horizontal gene transfer Within the same new generation genotypesphenotypes w a r39gi zjlif natural selectlon g L om 39 towr if good mutation those a Q survive and pass mutation 0 Wk 29 L52quot Unglr a 3415 RemJauztaor I 23th cut1amaole furr ww fquot a n o 0 rs t 0 cl I 0 rr 22 23 Microbial Species Since Bacteria and Archaea do NOT reproduce sexually the term species has a different meaning not defined as a interbreedingnatural population Microbial Species a collection of strains that share many stable properties and differ signi cantly from other groups of strains Microbial Strain subset of a microbial species a strain consists of the descendants of a single mmicrobial culture from single cell one strain is designated as the Mstrain 0 Permanent example usually one of the first strains of a species studied often most fully characterized not necessarily the most representative member of species 24 Naming Microbes Binomial nomenclature Genus and species Escherichia coli Bacillus subtilis Micrococcus lutcus Streptococcus lactis Staphylococcus aurcus T rcponcma pallidum Saccharomyccs ccrcvisiac GIIA 09 039 SCIINCE FATH 39 0 Cussm ATIQN V A anquot fw I 39 1915 Question of the Day Which is NOT true AMicroorganisms are more numerous than any other kind of organism BMost cellular microbes live independently as cells or cell clusters CMicroorganisms include bacteria archaea Protists and some fungi DViruses are not cellular they are not considered a microorganism Discovery of Microorganisms Robert Hooke Antony van 1665 Leeuwenhoek 1674 1676 described the fruiting structures of mold first to observe and accurately describe bacteria l of Mi rrrrrrrrrrrrrr la 26 ID BullzlnnIComls Copyright The McGrawHill Companies Inc Permission required for reproduction or display O I o 0 b 2 O 0 Lens o390 5 o 39 o Specimen 39 00 holder 0quot Q 0 FOCUS screw Handle b In oOIun C 1 39 wo t K 7 1 L 39 0 o O quot iuooooeuounu 39 witf quot N39lt my T D Brock Figure 19b Brock Biology of Microorganisms 11e Figure 19a Brock Biology of Microorganisms llle Pea r50 Prentice 3 II In C D 2006 Pearson Prentice Hall Inc 27 The Conflict Of Spontaneous Generation spontaneous generation living organisms can develop from nonliving or decomposing matter Francesco Redi 28 1668 discredited spontaneous generation for large animals 39 I 3 y 21 39 339 CityShirt Whot Tm Mica 8 V x Flask unsealed Flask cooled Flask covered with gauze Spontaneous Generation Disproved Louis Pasteur 1861 Copyright the McGrawHill Companies lnc Permission required for reproduction or display g 5 Microbes being destroyed it Vigorous heat is applied 29 Broth free of live cells sterile Neck on second sterile flask is broken growth occurs Neck intact airborne microbes are trapped at base and broth is sterile John Tyndall 1820 1893 Irish philosopher demonstrated that dust carries microorganisms provided evidence for the existence of exceptionally heat resistant forms of bacteria 30 Final Blows to Spontaneous Generation Ferdinand Cohn 18281898 discovered bacterial endospores ALSO classified bacteria based on shape used the genus name Mfor the first time 31 The Role of Microorganisms in Disease infectious disease believed to be due to supernatural forces establishing connection depended on development of techniques for studying microbes once established led to study of host defenses immunology 32 Indirect Evidence for the Germ Theory of Disease 1 39 f a I x L 3 n i a I i 0 Joseph Lister 1827 1912 Admired Pasteur Father of Modern Surgery developed a system of surgery designed to prevent microbes from entering wounds his patients had fewer postoperative infections amp Ignaz Semmelweis 1847 handwashing to prevent childbed fever 3 i k 2 if v 391 I J 391 quott 3 i 1quot v V R c Direct Evidence Robert Koch 1884 established the relationship between Bacillus anthracis and anthrax used criteria developed by his teacher Jacob Henle 18091895 these criteria now known as Koch s postulates still used today to establish the link between a particular microorganism and a particular disease 33 KOCH39S POSTULA I39ES quot 1 quot quot Healthy animal animal II39he suspected pathogenic Red f organism should be present blood gse we BEBEquot In all cases of the disease 9quot tissue under the les I39d Ibsen from Suspected m39croscope cell animals pathogen I f39 uzy 95 77 A 23939he suspected organism a 39s akagafp39ate No should be grown in pure Colonies o Lmsg zre organisms quoturm suspected 3 quot diseased or present health animal i A 39 pathogen I y 39lnocu l39a39te39healyt h y ani ma39l39Wiityl39i cells of suspected pathogen 3Cells from a pure culture of the suspected organism should cause disease in a healthy animal I 1 39R e39hseve39sleea quotar39itiggu e sample and observe by microscopy 439I39he organism should he taBOrato39ry Pure culture reisolated and shown to he Suspected culture must be the same as the original pathogen ix 2 Same organlsm as before Figure 1 1 2 Brock Biology of Microorganisms 1 We 2006 Pearson Prentice Hall Inc 34 Suppliestechniques used by Koch Agar way to grow culture on solid surface Petri dish nutrient broth and nutrient agar methods for isolating microorganisms So that you can get a pure culture 35 Developments in Immunology Edward Jenner 1798 used a vaccination procedure to protect individuals from smallpox NOTE this happened before the germ theory of disease was established Pasteur and Roux 1880 discovered that incubation of cultures for long intervals between transfers caused pathogens to lose their ability to cause disease attenuation Pasteur and his coworkers 1885 developed vaccines for chicken cholera anthrax and 36 rabies Developments in Industrial Microbiology Louis Pasteur 1856 demonstrated microorganisms carried out fermentations developed the process of pasteurization Alexander Fleming 1929 discovered penicillin Syrindipidis discovery 37 Developments in Microbial Ecology Sergei Winogradsky 18561953 Martinus Beijerinck 18511931 pioneered the use of enrichment cultures and selective media studied soil microorganisms and discovered numerous interesting metabolic processes such as nitrogen xation Nitrogen xation takes nitrogen from the atmosphere to make ammonia Winogradsky discovered chemolithotrophy Anarobic nitrogen xation Beij erinck Father of virology 38 Aerobic nitrogen xation 39 Question for 11615 The field of deals with the spread of disease in populations while the field of deals With an individual s response to disease AImmunology medical microbiology BPublic health immunology CMedical microbiology public health DMicrobial physiology molecular bioogy 4o Developments in Molecular Microbiology Avery MacLeod and McCarty 1944 DNA is the genetic material At first they thought DNA was protein because of the different typescombinations of amino acids Arber and Smith 1970 restriction endonucleases Endonucleases allows you to cut DNA Jackson Symons and Berg 1972 first novel recombinant molecule Where they cut dna and then pasted it back together They did plasmid the extra chromosomal pieces of DNA Woese and Sanger 1977 DNA sequencing Kary Mullis 1983 PCR polymerase chain reaction We live in the Age of Bacteria as it was in the beginning is now and ever shall be until the world ends Stephen Jay Gould 1993 Archaea Eukarya Bacteria Genetic 1DNA sequencing PCR Over Leeuwenhgek Winogradsky genetic genetics of DNA code 2 Discovery of Moecuar genome 500 39 39 material Archaea microbial genomes Str nt9mrcin l Era of Molecular BiologyGeneral Microbiology M I quotquot Mi ml mo t Gonomrcs and Early Days Discovery Medical and General Microbiology Proleomics Figure 117 Brock Biology of Microorganisms 11e 2006 Pearson Prentice Hall Inc 41 Chapter 21 24 The Study of Microbial Structure Microscopy and Specimen Preparation The Light Microscopes two types of light microscopes used in microbiology Brightfield Dark field light microscopes are compound 2 or more lens upper limit of magnification ISOOX Ours are only lOOOX because our objective lens is 10X upper limit of resolution 2 pm Lenses and the Bending of Light light is refracted bent when passing from one medium to another Refractive index a measure of how greatly a substance slows the velocity of light direction and magnitude of bending is determined by the refractive indices of the two media forming the interface bending depends on thickness Lenses Copyright TTTTTTT awHi lllllll nies Inc Permissi ooooo ui rrrrrrrrrrrrrr t i oooo display focus light rays at a n the focal point F F I distance between center of lens and gt J focal point is the i 393 focal length strength of lens 39 shorter focal length I W greater magnification The BrightField Microscope dark image against a brighter background has several objective lenses parfocal if you get it in focus for the lower power then it will stay in focus for the higher power Parcentric It s going to stay in the center Get the object in the center so that when you increase power it stays on the field total magnification Magnification of ocular lens X magnification of objective lens Copyright The McGrawHill Companies Inc Permission required for reproduction or display gt lnterpupillary adjustment Ocular 7 eyepiece Body assembly Nosepiece Arm Objective lens 4 Mechanical stage C oarse 7 adjustm Substage condenser Fine foc Aperture diaphragm control adJUStm a Stage a Base With light source K Field diaphragm lever Light intensity control Courtesy of Leica Inc Microscope Resolution 39 ability of a lens to separate or distinguish small objects that are close together clarity 39 Wavelengmof light used is major factor in resolution shorter wavelength i greater resolution so it s better to use blueViolent lters so that you can have a shorter wavelength Copyright The McGrawHill Companies Inc Permission required for reproduction or display W Objective Working distance 9 V Slide with l l I specimen Numerical aperture NA measure of light gathering ability NA is on the side of the microscope higher numerical aperture 2 greater resolution Copyright The McGrawHill Companies Inc Permiss required for reproduction or display A Q a Slide Air V e OilCover 39 glass 39 Increase refractive index 9 Increase NA which results in a greater resolution Copyright The McGraw Hill Companies Inc Permission required for reproduction or display Table 22 Properties of Objective Lenses 0 BJ ECTIVE I Property Scanning Low Power High Power Oil Immersion I Magni cation 4gtlt 10x 40 45 x 9o1oogtlt I Numerical aperture 010 025 055 065 1 25 14 I 0 Approximate focal length f 40 mm 16 mm 4 mm 18 20 mm l Working distance 1720 mm 4 8 mm 05 07 mm 01 mm I Approximate resolving power 23 pm 09 pm 035 pm 018 pm I L with light of 450 um blue light Should be 450 nm The DarkField Microscope image is formed by light re ected or refracted by specimen produces a bright image of the object against a dark background Light will come from the sides of the specimen instead of light coming up used to observe living unstained preparations May want to observe living organism to determine motility used to identify bacteria used to observe internal structures in eukaryotic microorganisms 1O Copyright The McGrawHill Companies Inc Permission required for reproduction or display 11 Objective ti Spechnen Abb condenser gt4 E j J Darkfield stop Copyright The McGrawHill Companies Inc Permission required for reproduction or display Charles StrattonNisuals Unlimited 12 Question 12115 0 Which one of the following parameters could be decreased to increase the resolution of a microscope a Working distance b Wavelength of light c Numerical aperture 1 Focal length e A and B f C and D Electron Microscopy Copyright The McGrawHill Companies Inc Permission required for reproduction or display Electrons replace light as the O O O R fl h lllumlnatlng beam 31205 So we really shorten the Range of electron microscope wavelength Colonial alga Pediastrum 10m 9 Red blood cell 2 a RIC6mm bacteria Rodshaped bacteria 0 Coccusshaped bacterium Amoeba Wavelength of electron beam is shorter than light 9 higher resolution Nucleus i l Wh39 bl d ll Magnlficatlon re oo ce 2 100000 X Resolution limit 1pm Esche ich aw l Staphy ococwsi Mycopasma bacteria 9 Poxvirus 5 nm this is 1000X greater quot 10 m AIDS virus resolution Poliovirus 10 nm 100 A Proteins 1 nm Amino acid 10 A small molecule 01 nm 1 A O Hydrogen atom Scanning gt tunnehng microscope 1transmission lllllllllllllllillllllllllllllILLll l i i i ii 21EScanning Copyright The McGraw Hill Companies Inc Permission required for reproduction or display Table 25 Characteristics of Light and Transmission Electron Microscopes Feature Light Microscope Transmission Electron Microscope Highest practical magni cation About 10001500 Over 100000 Best resolution 02 pm 05 nm Radiation source Visible light Electron beam Medium of travel Air High vacuum Type of lens Glass Electromagnet Source of contrast Differential light absorption Scattering of electrons l Focusing mechanism Adjust lens position mechanically Adjust current to the magnetic lens Method of changing magni cation Switch the objective lens or eyepiece Adjust current to the magnetic lens L Specimen mount Glass slide Metal grid usually copper Copyright The McGrawHill Companies Inc Permission required for reproduction or display Copyright The McGraw Hill Companies Inc Permission required for reproduction or display Photosynthetic 39membrane vesicle 14 George J WilderNisuals Unlimited Copyright The McGrawHill Companies Inc Permission required for reproduction or display AccV Spot Magn Det WD Exp 2pm 300 kV 30 15549x SE 74 O jhc CDCJanice Carr F RTurner 1 5 39 39 Figure 41 Ob Brock Biology of Microorganisms 11e 2006 Pearson Prentice Hall lnc Preparation and Staining of Specimens Why increases Visibility of specimen accentuates specific morphological features Preserves specimens Fixation preserves specimen structures and fixes them in position organisms usually killed by heats and stains and firmly attached to microscope slide Heat fixation routine use with bacteria and archaeons preserves overall morphology shape but may distort appearance of internal structures chemical fixation used with larger more delicate organisms can preserve morphology and internal structures 17 18 Spread culture in thin film over slide Figure 43 part 1 Brock Biology of Microorganisms 11e 2006 Pearson Prentice Hall Inc Flood slide with stain rinse and dry Pass slide through flame to fix II Heat fixing and staining Figure 43 part 2 Brock Biology of Microorganisms 1 He 2006 Pearson Prentice Hall Inc Dyes Stains 39 make internal and external structures of cell more Visible by increasing contrast with background have two common features Chromophore groups chemical groups with conjugated double bonds give dye its color ability to bind cells 19 Types basic dyes dyes with positive charges bind to negative charged structures acidic dyes dyes with negative charges bind to positive charged structures Copyright The McGraw Hill Companies Inc Permission required for reproduction or display Table 24 Ionizable Dyes Type of Dye Examples Basic dyes Methylene blue basic fuchsin crystal violet safranin malachite green Acidic dyes Eosin rose bengal and acid fuchsin possess groups such as carboxyls COOH and phenolic hydroxyls OH 20 Characteristics 1 Have positively charged groups bind to negatively charged molecules such as nucleic acids many proteins and the surfaces of bacterial and archaeal cells In their ionized form have a negative charge and bind to positively charged cell structures Simple Stains 39 simple staining a single staining agent is used to determine shapesize Copyright The McGrawHill Companies Inc Permission required for reproduction or display a Crystal violet stain b Methylene blue stain of Escherichia coli of Corynebacterium a Kathy Park Talaro b Harold J Benson 21 Differential Staining 39 use more than one dye to preferentially stain features used to detect presence or absence of structures 39 divides microorganisms into groups based on their staining properties 1 Gram stain 2 Acid fast stain 23 Gram Staining discovered 1884 by Christian Gram Copyright The McGrawHill Companies Inc Permission required for reproduction or display most widely used differential staining procedure divides bacteria into two groups based on differences in 11 wall structure Grampositive purple Gramnegative Pinkred 0000 0000 0090 0009 Steps in Staining Step 1 Crystal violet primary stain for 1 minute Water rinse Step 2 Iodine mordant for 1 minute Water rinse Step 3 Alcohol decolorizer for 10 30 seconds Water rinse Step 4 Safranin counterstain for 30 60 seconds Water rinse Blot dry State of Bacteria Cells stain purple Cells remain purple Grampositive cells remain purple Gramnegative cells become colorless Grampositive cells remain purple Gramnegative cells appear red Copyright The McGrawHill Companies Inc Permission required for reproduction or display a C Perhmgens 2 y 39 x x t I o o 39 w h A 39 r o v 5 I I f r 539 f iquot it 39 CI E can d N gonorrhocao Diplococci 24 25 AcidFast Staining particularly useful for staining members of the genus mycobacterium high lipid content in cell walls mycolic m is responsible for their staining characteristics 26 Staining Specific Structures endospore staining heated double staining technique bacterial endospore is one color and vegetative cell is a different color capsule staining Negative stain capsules colorless against a stained background agella staining Mordant applied to increase thickness of agella This helps so that the agella can be seen Flagella aids in mobility Copyright The McGrawHill Companies Inc Permission required for reproduction or display Copyright The McGrawHill Companies Ina Permission required for reproduction or display Lgt a 5 W1 39 u39 JJA9 11 A I o v 39 f irc wnra y 39 v z z r vquot39 r pw z quot3 quot39 quot quot1351 39l v A quot 1 v H a Gram Stain c Endospore stain of Bacillus Purple cells are gram DOSthe subtilis showing endospores RGd CEIIS 61r 9 gram negathe red and vegetative cells blue Hamid FiSher Manfred Ka ePeter Arnold Inc 9 39 d Capsule stain of bacteria Steven P Lynch Copth O The McGrawvHill Companies Inc Permission required lor reproduction or display b Acidfast stain Red cells are acidfast e Flagellar stain of Proteus vulgaris A basic stain was used to Blue cells are non acnd fast coat the agella 27 0 David B Fankhausar Unwaraily 01 Cincinnati Clormont Colloga httpIbiologycicucodulFankhausorl Bacterial Cell Structure and Function Size Shape and Arrangement 5 u 39 39quot1 C2115 cocci and bacilli rod shaped most common prokaryotes Norbert Pfennig Rad Norbert Pfennig arrangement determined by plane of division and degree of separation after they divide Norbert Pfennig some don t completely ddsmamaquot k E appendaged lelde bacteria size Hlaamentums varles bacteria Figure 41 quotIll Brock Bile91y of Micmnarganisms l 1 IE I323 Z Pearaam Prentice Hall ilnc cocci s coccus spheres Cocci Copyright The MoGlraWIHIiII Companies lino Permission required for reproduction or dispiay diplococci s diplococcus pairs Like Where they don t completely separate in division Photo Researchers Inc streptococci chains Copyright The MoGrawHilll Companies lino Permission required for reproduction or disp iay staphylococci grapelike clusters tetrads 4 cocci in a square sarcinae cubic configuration of 8 cocci Bruoe Iverson Copyright The MBGFEIWlllII Companies Inc Permission required for reproduction or display Bacilli bacilli s bacillus rods coccobacilli very short rods Copyright The McormsTilI E nb hr fi T Fr i si h39l i i himr reproduction or display H l Vquot n P Vibrios resemble rods comma shaped spirilla s spirillum rigid helices spirochetes exible helices c Leprospira interrogans a spirochele CDCINClDlHPIJanioe Carr Other Shapes and Arrangements filamentous mycelium network of long multicellular laments pleomorphic Variable in shape Usually happens because there isn t a cell wall so it s more exible Archaea pleomorphic branched flat square other unique shapes Copyright The MoGrawliill Companies Inc Permission required for reproduction or rzlispie39gim3 MCGWW mamas 39 Pemm Wire f mm rdisp39a e Hypnomiorobiorn d Streptomyoes a filamentous bacterium Dr Amy Gehring The Shorter Bergreys Marl ual of Determinations Bacteriology Bie John G Holt Editor I Brergef39s Manual Trust PubFished by Williams S Wilkins Baltimore MD Copyright The MoGrawHiil Companies Inc Permission required for reproduction or display WWW 39339 WWWHi Companies 39quotC Permission requi39ed rm FWOd L39Etion display E r 1 From Walther Sioeclienins Walsby39s Square Bacterium Fine Structures of an Orthogonal Proceryore From JILTJ Staley MP Bryant N Pfenning and JG Holt Eds Bergey s Manual of System stir Bacteriology Vol 3 1939 Williams and Wilkins Co Baltimore Robinson Dept ofitilicro U ofCeI LA Size COpyrigm The MCGWW39HWI C U39mpameS Inc Permission required for reproduction or display Specimen Appraximate diameter or W39iidt39hi x lienigiith Cell Slze Ranges 1 quotm Oscillatorie Eukaryotes Red bliocd cell 73000 08 pm hundreds of um ccii 1300 x 4000 BacteriaArchaea gum 750 um Streptococcus 8004 51000 Viruses iPOXVirUie 230 x 320 In uenza virus 85 03901 Hm 1 Hm T2 Eccii bacteriophage 65 x95 Tobacco mosaic virus 1i 5 x 300 Policmyelitis virus 27 Table 41 Cell size and trelume ef39 relkaryetic cells from largeat te the smallest Organiem Characteristics Merpheiegy 5235 Mm Ce valume Mn3 E celi volumes Thiemargarita aamiiaierteie Sulfur chemelithetreph Ceccl li39i chaine 7540 20030003000 ma Epulepieciurrt tiaheleeni Beggiatea 5p Achrematium eaaiiferum Lyngbya majuscul Prechleren Sp Thievulum m39ajua Staphyiethermus marinas Titaneseleirillum eelea Mageetebacterium bawaricum Escherichia cell P39elagibacter ubiqu e Chemeergartetreph Suliur chemelithetreph Sulfur chemelithetreph Cyanebacterium P rec h lie rep hyte Sulliur chemelithetraph Hyperthermephile Suliur chemelithetreph Maghetetactic bacterium Chemeerganetreeh Marihe chemerganetreph Reds with tapereel encle Filaments Ccci Filaments Cecci Ceccl Cecci in irregular clusters Carved recle Reds Reds Bede 8039 600 50 x 160 8 80 30 i8 15 5 X 30 2 l0 lgtlt2 029605 BfUU39UDUO l 000000 80000 400130 1 EWCJ39O 3000 1 300 6010 3 001 4 15 mt 4 1gtlt 10d 104 X 103 15 x 103 9 x 139 a gtlt 102 3915 Mycepiaema paeumeniae Pathegenic bacterium Pleemerphicb 12 005 aWhere eel ene namer quotIa given thie la the diameter ei spherical cells The ealuee given are iar the largest cell eiae ebeereeci hquot each species Fer example ier T namibieneie arr average cell is ehly abeut 2010 are in diameter Burt en eccaaiertr giant celle at EU arr are elaeertrecl Likewise art average cell cit marinas ie abeut i am irr diameter bitriyceplaeme is a cell wall leee bacterium and can take er many shapes Eelquot emerjehic rriearte marry ehapee l Source Data elata i39hecl irem Schulz Hth and BB allergeheeh 2001i 1 an Herr Micrebiel 1059131 Eeeyrighi e eaaa Peereen Etiueatiert Inc eubliehirtg as Feereert Eieniamin Cummings Surface area lmr2 126 pm Volumews42ma Surface 3 Volume i Surface area umz Volume pm3 15 Volume Figure 4131 Bruck Binl agy Elf Micmnrganismi 1 11m r i 2am Peawmm PramEa Halli Inc Cell Organization Capyright e The litiieGrawllill C ar npaniea l ne Permission required far reprecluetian er display Table 31 Summon Bacterial Structures and Their Functions Plasma membrane Selectively permeable laarrier mechanical lanainclarjir at cell nutrient and waste transpartg lcrcatien pf manyr metabolic precesses respiration photosynthesis detectian atquot entrian mental cues for chemptasis I Gas vacuele An inclusian that presides buveyancy fer eeting in aquatic envirnnments Ribasames ratein synthesis I inclusions Storage at carbun phosphate and other substances i Nuclepicl Lancalizatien pf genetic material DNA Periplasmic space in typical Gramenegative bacteria cantains hydralytic enzymes and binding preteins far nutrient pracessing and u pta ke in typical Gram pesitive bacteria may be smaller er absent l Cell wall Pratectien tram esmetic stress helps maintain celll shape Capsules and slime layers Resistance ta phagecytasis adherence ta surfaces Fimbri ae and pili Attachment to surfaces bacterial canjugatian and transformation twitching and gliding rnatilityF i Flagella Swimmin and swarming matilitjyF Enndespere Survival under harsh ensiranmental canclitians L 10 Copyright The McGrawiHiN Companies Inc Permission required for reproduction or display Plasma Capsule Ribosomes Calm walll membrane Nucleoid Fi mbriae Chromosome Inclusion FlageHum DNA Plasma Membrane encompasses the cytoplasm cytoplasmic membrane selectively permeable barrier interacts with external environment receptors for detection of and response to chemicals in surroundings Transport systems metabolic processes Membrane Structure 39 lipid bilayer O mposedof ydiiza it39 PhOSpholipids ksquot a g i F o a a i a a quotN V 39l39 y HVdEOPhObiC 1 Fatty acids 39 amphlpathlc llpldS region 39 Imlar ends 39 Hydrophilic E 39 hydrophilic region 39 39 nonpolar tails f GHy em Pihosphate o Figure 414 Brock Biology of Microurganisms 1 He 93 2006 Pearson Prentice lilall Imci 12 Bacterial Lipids opyright The McGrawHill Companies Inc Permission requured for reproduction or display 0 saturation levels of membrane lipids re ect environmental conditions bacterial membranes Sagan lack sterols but do contain sterollike molecules hopanoids stabilize membrane 23H Suppose you have discovered a microorganism that is 7 um in diameter Which one of the following hypotheses would be the best to propose a the microbe is a Virus b The microbe is a bacterium c The microbe is a eukaryote d The microbe is a Virus 21 bacterium 15 Fluid Mosaic Model Copyright The McGrawHilll Companies Iine Permissien required for reproduction or display Oli39gosaecnaride gra quot quotf Integral quot 39 V V protein Integral gt V protein Glyee ipid Hydrophobic 16 Membrane Proteins peripheral loosely connected to membrane comprise 2030 of the membrane proteins integral embedded within membrane amphipathic comprise 7 080 of the membrane proteins carry out important functions transport secretion Energy conservation 17 Bacterial Cell Wall Rigid structure that lies just outside the plasma membrane contains peptidoglycan Only found in Domain Bacteria Very protective and selective to What can come into the cell 0 Items smaller than 2 um functions provides shape to cell protects from osmotic lysis may contribute to pathogenicity protects from toxic substances Copyriglii Tlie M CGiElWi il Companies Inc Permission required for reproduction or display Cell Walls of Bacteria Bacteria are divided into two major groups based on the Gram stain differential staining due to cell wall structure quyright The McGrm Hill E Q LtiiirrrlFIalrnirs395z lm Permi ain required fur reprndluctian Di39 display Dapyri mmmmm McGrawlli lllllll les llnc Permissicm requi mmmm r reprodtmhion or display Tr Dalila ame magma l g o iagnaa 9 eisgian39me ll drarWMomdisplay T39h e gra39miln ega39tl v e Ge M The gramposltlve cell wall Cell lPeptidloglycan WEI H lPlasma mlelmbra e Peptldoglycan Plasma membrane N I w Periplasmic a Cell wall space 39veri e Bina ice hots Service 7 121 Emi gef ialngica Phams ewim 18 purple g Name That Wall lPeptidogiycan Cytopiasmic membrane L Pater Figure all2 Brock Biology of Microorganisms 1 1 are 2006 Pearson Premtice Hell 11C Grampositive 19 Outer membrane Cytoplasmic membrane Peyptidoglycawn 7 Figure 42dl Brock Biology of Microorganisms 1 1 fe 2006 Pearson Prentice iIalli IIrIc gramnegative k and S F Caint T D Bro 4 Peptidoglycan Structure important component of both gram positive and gramnegative bacteria Even though it s thin in gramnegative it helps to protect the cell and give it shapestructure meshlike polymer two alternating sugars form backbone N acetylglucosamine gNAGz N acetylmuramic acid NAM 20 alternating D and L amino acids NAteiylluwsmine G I Fl Na cetyll mup CH3 JI e 39 a h sensitive quot39 quotI u I g J II Peptide crrossallinks 3 awn39 NH rAlunime 1 w M l 59 VETS TCE39LECH TCOOH I quotNquot NH 8 na iwiumicm W V i I Mesadimmin i I I a H39 1quot NHI plmellhm mud I i 1 a 3913E ff SOP ni lmmine F Figure 429 Brack Bi l giy39 if Micrnaurgani ms 1 HE 21 I21 I 2006 Pearmm Pmntite all lint Peptidoglycan chains are crosslinked by mtides for strength D rig nia39T e McGr aw Hilm Camp u rq gm 7 at in urd iigsplay E 60 direct linkage S aureus indirect w w by Gm i u i 22 copyright The M CGrawHi ll Companies Inc Permission required for reproduction or dismay Peptide 3 Side chain EN AG Polysaccharide backb ne 23 Figure 4 317 BI fk Bial ngw sf Micm39u ranisms 114 s IE Z Pearsam Prentice Hall Im GramPositive Cell Walls composed primarily of peptidoglycan up to 90 of wall may also contain large amounts of teichoic acids negatively charged maintain structure of cell envelope protect from harmful substances may bind to host cells 24 pathogenic bacteria lipoteichoic acids attached to membrane some grampositive bacteria have a layer of proteins on surface of peptidoglycan Grampositive Pepti dog lycan UNIMembrane 25 Capyrighi The McGrawHill Campaniasj MG Permissian reuired for rapro F r Lipoteicoic acid Teichaic acid Pepticiczgly39tS an I erfipilasmic space Plasma membrane uctiar39m or display 26 GramNegative Cell Walls consist of a m layer of peptidoglycan surrounded by an outer membrane outer membrane composed of lipids lipoproteins and popolysaccharide LPS m teichoic acids Gramnegative Peptidoglycan Membrane I39D IT plasm quot Outer memlarane llpopolysaccharide and protein Figure 427b Brock Biology of Microorganisms 1 We 2006 Pearson Prentice Hall Inc 39 1 27 GramNegative Cell Walls peptidoglycan up to 10 of cell wall Periplasm may constitute 20 40 of cell volume many enzymes present in periplasm hydrolytic enzymes to break down food transport proteins and other proteins Outer membrane lies outside the thin peptidoglycan layer Braun s lipoproteins connect outer membrane to peptidoglycan Copyright The McGrawiHili Companies in Permissron required for reprodoction or dispiay i3 r r r r 7 7 i r r f r r Braqu 1 or r r inoprotern Outer membrane as 1119 1 i 7 I 7 3 w quotw 1w 539 aquot 9 39 quot Peri I quot pasrnlo spaceend peptidogiyoen Piesnna membrane Integral protein 28 Lipopolysaccharide LPS three parts lipid A 0 Extremely important core polysaccharide 0 side chain 0 0 antigen because it will be recognized as foreign in the body 0 0 speci c polysaccharide Lipid A embedded in outer membrane core polysaccharide amp 0 side chain extend out from the cell I Itspecific polysaccharide H Core polysaccharide N Lipid A l 3 E p o N p I Hep p KDO I a p o n P 29 Comgm ti 2209 Pearson Eeu calion me publishing as Pearson Benjamin Cummings 30 Copyright The MoGrawHill Companies inc Permission required for reproduction or display Friaran Abe Rho I a n Main Abe Silo MAG I IEiaI islc Gal Hep q E 0 side chain V 7 Hop ethanoiamine KDD I 7 7 7 KDD KDO ethanolamine GioN GioN go a Fatty acid C re pDIyS ccharide J I Lipid A 39 a i r39 it From NI Kastowsky T Gutherlet and H Bradaozek Journai of Bacteriology 774498 4806 1992 31 Importance of LPS contributes to negative charge on cell surface core polysaccharide helps stabilize outer membrane structure lipid A may contribute to attachment to surfaces and biofilm formation Biofilm assemblage of bacteria that makes a matrix that helps them stick together creates a permeability barrier may mutate to protect from host defenses 0 antigen can act as an endotoXin poison lipid A Causes gastrointestinal distress vomitdiarrhea 32 GramNegative Outer Membrane Perm more permeable than plasma membrane due to presence of porin proteins and transporter proteins porin proteins form channels through which small hydrophilic molecules like sugars can pass eab y I quot 39 39 1 Equot w a leim 39m E II I I 39 I 1 o Ir 39 V 391 r u V 39 quot I l IP V p r i I s a 7 39 r 7 WI 39 i r 39 l I a quot a In E V Ll 39 M J 2712 Georg E Schulz Comparison of GramPositive and GramNegative Cell Walls Lipoteichoic acid Teichoic acid a 1 l 39 N U Braun s l lipoproteirl II V l 3 l g lt wLipopolysaccharide B 1 g I D Outer membrane Periplasmlc space Periplasmlc a gt Space and g peplidoglycan m L Q lt E E E a n E Plasma Phosphollpid Peptidoglycan lemma lnlegral protein 33 Question of the day If you only had access to the cell wall of a bacterium how would you determine if it was gram or gram 0 B look for the presence or absence of teichoic acids 0 C Look for the presence or absence of lipopolysaccharides Osmotic Protection Hypotonic environments SOIUteoutside cell lt SOIUte inside cell water moves into cell and cell swells cell wall protects from lysis Lysislysing breaking open Hypertonic environments SOIUteoutside cell gt SOIUteinside cell water leaves the cell Plasmolysis occurs Cell shrinks as water leaves 35 36 Evidence of Protective Nature of the Cell Wall Lysozyme breaks the bond between NAG and NAM Penicillin inhibits peptidoglycan synthesis if cells are treated with either of the above they will wit they are in a hypotonic solution This picture demonstrates a cell lysing during cell division Without A Cell Wall survival in isotonic environments protoplasts spheroplasts Mycoplasma no cell wall plasma membrane more resistant to osmotic pressure Copyright the McGrawHiii Companies inch Permission required for reproduction or diepiay Penicillin inhibition of wail synthesis Incubation in isotonic Eran ferio i SWGI ImgdUB e i medium quot pr on to H20 influx Lysis i m ecilu rn 39 h s it p i iProtoplast 37 Components Outside of the Cell Wall Capsules Slime Layers 39 usually CODIPOSed 0f similar to capsules except polysaccharides sugar well organized and not easily removed from cell easily removed protective advantages resistant to phagocytosis protect from desiccation drying out especially in a hypertonic environment They also eat from Within their capsule if needed to survive diffuse unorganized and slime may aid in motility exclude viruses and detergents END OF UNIT 1 The Cytoskeleton role in cell division protein localization and determination of cell shape Copyright The iMoGrawHiil Companies Inc Permission required for reproduction or display Table 32 Bacterial Cytoskeletai Proteins Function Comments Cell division Wideiv observed in bacteria and archaea i Btu bNBtol oB Uniknown Observed onlyr in Prosthecobocter sppt thought to be encoded by i eukaryotic tubuiin genes obtained by horieontai gene transfer t TubZ Possibiv plasmid segregation Encoded by large plasmids observed in members of the genus Bociiius i Austin Homologues MamK Positioning magnetosomes Observed in rnagnetotactic species MreBiMbl Helps determine cell shape may be involved in Most rodeshaped bacteria chromosome segregation locaiizes proteins i l ParM Plasmid segregation Plasmid encoded 1 intermediate Filament Homologues l CreS crescendo induces curvature in curved rods Eoiuiioba39ctercrescentus Unique Bacterial g ioskeieroi Proteins Mint Prevents polymerization of FtsZ at ceii pole Ma ny rodshaped bacteria Pam Segregates chromosomes and plasmids Observed in many species inciud39iing Vibrio choieroe Ci crescentus and Thermos thermophiius Inclusions Aggregatesstorage of Organic 0139 inorganic T a W i i material quot granules crystals globules some are enclosed by a singlelayered membrane or invaginations of plasma membrane Ralph A SlepeclkyNisuals Unlimited 0 Not organelles even though some have membrane Storage Inclusions Ep39ymml 39l39he Mu mHi Gompay l Inc Flammlmln required Emir mpm u lm urditsplajr storage of nutrients metabolic end products energy building blocks 0 Carbon glycogen poly hydroxybutyrate m2 Phosphate polyphosphate granules Sulfur globules Pop up if needed disappear if not Nitrogen cyanophycin granules REpmlmt d from Till Ehli l i EsmeH39s Manual mi Whaling Bantarlnln y Eu Jnlhni E Edwa 19 1 Berger39s Manual 39l39ms39l IFhuhlllshE h r 39H39u39llllnms E Williams Bahlrmna MD 42 43 P 21 2quotzclJ5fal39xlal st Val1 4 m Munnu may I 39 5L Line I Gasman15 nixMEI hum Tor Uaw 5uzl39u1nellr3lugt Jarun 2m mnmmzsucn I al Emm Heals provide buoyancy Cylinders With protein membrane that let gases in so that they can oat easier Floating is important because it helps them getting closer to sunlight if they photosynthesize Also oat higher and lower to find the perfect depth for proper oxygen levels and to find nutrients throughout the water Only found in aquatic Archaeans Other Inclusions 111a gnetosomes magnetite particles for orientation in Earth s magnetic field Can oreint themselves north and south They function to bring themselves down to the bottom of the water column lake Reason for this is to get to the nutrients that are found at the bottom of water in sediments Chapter 75 Laboratory Culture of Cellular Microbes Question 12315 After iodine is added in the Gramstaining procedure before the ethanol rinse Gram positive cells would appear and gram negative cells would appear a Purple pink b Pink purple c Purple colorless 1 Purple purple Culture Media nutrient preparations devised to support the growth reproduction of microorganisms Macronutrients Ll t 3quot Carbon hydrogen oxygen phosphorus nitrogen sulfur i SPONCH J Micronutriets 0 Unless you re using highly puri ed water you don t have to worry about adding micronutrients like zinc If water is too pure you don t get the trace metals that some organisms need to grow Inorganics growth factors Organics Vitamins can be liquid or solid solid media are usually solidified With agar Copyright Ihe Mcb rawHIII Companles Inc Permrssron reqUIred tor reproduction or display Table 74 Types of Media Basis for Classi cation Types Chemical composition De ned synthetic complex Physical nature Liquid semisolid solid Function Supportive general purpose enriched selective differential g a a a 4 Defined or Synthetic Media all components and their concentrations are known Copyright The McGrawHill Companies Inc Permission required for reproduction or display Table 75 Examples of De ned Media BG 11 Medium for Cyanobacteria Amount gliter Medium for Escherichia coli Amount gIiter NaNO3 15 Glucose 10 KZHPO4 3H20 004 NazHPO4 164 MgSO4 7H20 0075 KH2P04 15 l CaClz 2H20 0036 NH4ZSO4 20 Citric acid 0006 M9504 7H20 2000 mg 1 Ferric ammonium citrate 0006 CaClz 100 mg EDTA NazMg salt 0001 FeSO4 7H20 05 mg I Na2C03 002 Final pH 68 70 Trace metal solution1 10 mlliter J Final pH 74 Complex Media contain some ingredients of unknown composition andor concentration Copyright The McGrawHill Companies Inc Permission required for reproduction or display Table 76 Some Common Complex Media Nutrient Broth Amount gliter Tryptic Soy Broth Amount gliter MacConkey Agar Amount gliter Peptone gelatin 5 Tryptone pancreatic 17 Pancreatic digest 170 I hydrolysate digest of casein of gelatin 39 Beef extract 3 Peptone soybean digest 3 Pancreatic digest 15 I of casein Glucose 25 Peptic digest of 15 I animal tissue Sodium chloride 5 Lactose 100 i Dipotassium phosphate 25 Bile salts 15 I Sodium chloride 50 1 Neutral red 003 l Crystal violet 0001 I MacCo nkey Agar A93r 135 J coo Some Complex Media Components peptones protein hydrolysates prepared by partial digestion of various protein sources extracts aqueous extracts usually of beef or yeast agar sulfated polysaccharide used to solidify liquid media most microorganisms cannot degrade it Functional Types of Media general purpose media supportive support the growth of many microorganisms eX tryptic soy agar Or tryptic soy broth enriched media general purpose media supplemented by blood or other special nutrients eX chocolate agar Not actually made of chocolate It s boiled blood agar Minimal media contains the minimal necessities for growth of the wildtype only contains inorganic salts a simple carbon source and water selective media favor the growth of some microorganisms and inhibit the growth of others eX MacConkey agar selects for only gramnegative bacteria 10 Differential Media distinguish between different groups of microorganisms based on their biological characteristics eX blood agar Hemolytic versus nonhemolytic bacteria eX MacConkey agar Lactose fermenters versus nonfermenters Copyright The McGrawHill Companies Inc Permission required for reproduction or display B hemolytic colony a Blood agar Kathy Park Talaro Copyright The McGraw Hill Companies Inc Permission required for reproduction or display Table 77 Mechanisms of Action of Selective and Differential Media Medium Blood agar Eosin methylene blue EMB agar MacConkey MAC agar Mannitol salt agar 11 Functional Type Enriched and differential Selective and differential Selective and differential Selective and differential Mechanism of Action Blood agar supports the growth of many fastidious bacteriaThese can be differentiated based on their ability to produce hemolysins proteins that lyse red blood cells Hemolysis appears as a clear zone B hemolysis or greenish halo around the colony ahemolysis eg Streptococcus pyogenes a Bhemolytic streptococcus Two dyes eosin Y and methylene blue inhibit the growth of Grampositive bacteria They also react with acidic products released by certain Gramnegative bacteria when they use lactose or sucrose as carbon and energy sources Colonies of Gramnegative bacteria that produce large amounts of acidic products have a green metallic sheen eg fecal bacteria such as E coll The selective components in MAC are bile salts and crystal violet which inhibit the growth of Grampositive bacteriaThe presence of lactose and neutral red a pH indicator allows the differentiation of Gramnegative bacteria based on the products released when they use lactose as a carbon and energy source The colonies of those that release acidic products are red eg E CO A concentration of 75 NaCl selects for the growth of staphylococci Pathogenic staphylococci can be differentiated based on the release of acidic products when they use mannitol as a carbon and energy source The acidic products cause a pH indicator phenol red in the medium to turn yellow eg Staphylococcus aureus Isolation of Pure Cultures 39 pure culture population of cells arising from a single cell allows for the study of single type of microorganism a mixture of cells is applied to an agar surface so that individual cells are well separated from each other streak plate spread plate and pour plate The Streak Plate 39 technique of spreading a mixture of cells on an agar surface with an inoculating loo 39 each cell can reproduce to form a separate colon1visible growth or cluster of microorganisms Aseptic Transfer 0 b 13 Copyright The MoGrawHill Companies Inc Permission required for reproduction or display Note This method only works if the spreading tool usually an inoculating loop is resterilized after each of steps 1 4 Confluent growth at beginning of streak Isolated colonies at end of streak Figure 54c Brock Biology of Microorganisms 1 1 e 1 5 Figure 5 dab Brock Biology of Microorganisms 11e 2006 Pearson Prentice Hall Inc 2006 Pearson Prentice Hall Inc James A Shapiro University of Chicago Spread Plate and Pour Plate spread plate small volume of diluted culture is transferred to agar surface culture is spread evenly over surface with a sterile bent rod hockey stick pour plate Diluted samples are mixed With liquid agar mixture of cells and agar are poured into sterile culture dishes both provide isolated colonies opportunity to enumerate the bacteria in a sample 16 countable plate 30300 colonies Spread Plate Copyright The McGrawHill Companies Inc Permission required for reproduction or display A small amount of the sample is pipetted to the center of a solidified medium The spreader is cooled and then used to spread the sample evenly over the surface of the medium The glass spreader is sterilized by dipping it into ethanol and briefly flaming it a 17 Kathy Park Talaro Pour Plate Copyright The McGrawHill Companies Inc Permission required for reproduction or display The original sample is diluted several times 10 ml 10 ml 10 ml 10 ml Original 9 ml H20 9 ml H20 9 ml H20 9 ml H20 sample 101 dilution 1 02 dilution 103 dilution 104 dilution Some of the dilutions often the 1 0 m39 1 0 m39 most dilute are mixed with warm agar and poured onto the plates Isolated cells grow into colonies on the surface appear round and within the medium appear lensshaped The isolated colonies can be counted or used to establish pure cultures 18 Copyright McGrawHill Which one of the following choices best describes a culture medium with the following characteristics only allows for the growth of Grampositive bacteria supplemented with soil and contains yeast extract mission required for reproduction or display a Selective enriched complex b Differential general purple defined c Selective differential general purpose d Enriched minimal synthetic Microbial Growth on Solid Surfaces species form ity of Chicago 39 differences in growth rate from 3d ges to center are 123t9rr am1we due to James A Shapiro Univers nutrients and toxic products cells may be in some areas 20 James A Shapiro University of Chicago 21 Copyright The McGraw Hill Companies Inc Permission required for reproduction or display Copyright The McGraw Hill Companies Inc Permission required for reproduction or display Dr Eshel BenJacob Image courtesy Mark Schneegurt Copyright the McGrawHill Companies Inc Permission required for reproduction or display Punctiform Circular Filamentous Irregular Rhizoid Spindle Flat Raised Convex Pulvinate Umbonate Entire Undulate Lobate Erose Filamentous Curled Microbiology Exam 1 Review in class Practice Dilution Problems 1 a 139 x 106 bacteriaml i Do not take into account ofthe colonies that are not countable not between 30300 ii Therefore 139 is the only countable one b No 2 10396 3 4 ml shake 12 ml dHZO a Take a ofthe 16 ml to get 4 ml of shake So then the rest is going to be 12 ml Exam Review Microscopes know how to increase resolution whats refractive index focal length focal point with magnification 0 Know the bright field dark field doesn t change it much change light source why might you wanna use it transmission electron microscopes why do you get better resolution decrease the wavelength to increase resolution 0 Evolution timeline don t have to know numbers know the general idea of the order and anything significant oxygen first so what was the first oxygen bacteria cyanobacteria o How do we study evolution fossil records how we ve gotten the evidence on the timeline o Endosymbiosis 0 Tree of life be able to read it and know what it means I All it is telling you is relatedness based on small subunit ribosomal DNA I Length of line tells how close they are I At the base of tree Luca last universal common ancestor 0 Know braches and Archaean are more closely related to eukaryotes even though they look like bacteria 0 Labeling cell membrane and cell walls staining 0 Bacteria shapes 0 Don t get cell wall mixed up with cell membranes 0 Same pictures from presentations 0 50 multiple choice 0 Different types of media Gram stains purpose what they show 0 History of micro concentrate on the big guys Not the bunches on one slide o If he s by himself or two together know no dates though o Coch s postulates and different techniques he used for postulates 0 Plate count 0 Know range sizes 0 Name tells shape of bacteria 0 Know diseases but don t have to know specific examples like in pictures 0 Know different of gram and in a picture Copyright The McGraw Hill Companies Inc Permission required for reproduction or display Organisms and biological entities studied by microbiologists I can be includes includes eg eg eg eg composed of composed of composed of composed of Yeasts Algae Escherichia Methanogens Molds Protozoa coli Slime molds Cytoplasmic membrane Endoplasmic reticulum Ribosomes Nucleus Nucleolus Nuclear membrane Golgi Cytoplasm Mitochondrion Chloroplast I 10 um Cytoplasm Nucleoid Ribosomes Plasmid l l 05 pm Cytoplasmic Cell wall membrane Figure 21 a Brock Biology of Microorganisms 11e 2006 Pearson Prentice Hall Inc
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