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MICROBIAL NUTRITION,
GROWTH, MEASUREMENT, AND CONTROL
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RESOURCES ON LINE• http://www.bact.wisc.edu/Microtextbook/i
ndex
• AN ONLINE TEXTBOOK
• http://www.bact.wisc.edu/bact100/KimballLinks.html
• http://science.nhmccd.edu/Biol/microbio.html
• http://science.nhmccd.edu/BioL/animatio.htm
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MICROBIAL NUTRITION
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MICROBIAL HABITATS
• OCEANS
• ANTARCTIC ICE CAP
• VERTEBRATE AND INVERTEBRATE GUT
• JAMS, JELLIES, PICKLES
• VIRTUALLY EVERYWHERE
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MICROBIAL NUTRITION
• ORGANISMS MUST GET FOOD FROM ENVIRONMENT
• WIDE VARIETY OF NUTRITIONAL STRATEGIES
• http://www.mansfield.ohio-state.edu/~sabedon/campbl27.htm#photoheterotroph
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NUTRITIONAL TYPES• PHOTOAUTOTROPHS• CHEMOAUTOTROPHS• PHOTOHETEROTROPHS• CHEMOHETEROTROPHS• http://72.14.203.104/search?
q=cache:EkMmMJD4ZtIJ:rcw.raiuniversity.edu/biotechnology/BTechbiotech/microbesandus/lecture-notes/lecture-10.pdf+microbial+nutrition+chemoheterotroph+photoheterotroph+photoautotroph&hl=en&gl=us&ct=clnk&cd=5
• http://www.sinauer.com/perry/Perry_SG_05.pdf
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PHOTOAUTOTROPHS
• LIGHT AS ENERGY SOURCE• CARBON DIOXIDE AS CARBON
SOURCE• H2O, H2, OR H2S AS ELECTRON DONOR• http://72.14.203.104/search?
q=cache:sclpkxyy_VMJ:www.homepage.montana.edu/~umbls/mb301/lecquiz2key.pdf+photoautotroph&hl=en&gl=us&ct=clnk&cd=26
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OXYGENIC PHOTOSYNTHESIZERS
• OXYGEN GIVEN OFF• GENERATED FROM WATER• PHOTOSYSTEM II PRESENT• CYANOBACTERIA, ALGAE, AND
GREEN PLANTS• http://72.14.203.104/search?
q=cache:3iUlq0IL7tsJ:meetings.copernicus.org/egu2005/download/EGU05-J-11198-1.pdf+oxygenic+photosynthesizer&hl=en&gl=us&ct=clnk&cd=5
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ANOXYGENIC PHOTOSYNTHESIZERS
• DO NOT USE WATER• USE H2S, H2 , SULFUR OR SULFIDE• USE BACTERIOCHLOROPHYLLS• http://72.14.203.104/search?
q=cache:3iUlq0IL7tsJ:meetings.copernicus.org/egu2005/download/EGU05-J-11198-1.pdf+anoxygenic+photosynthesizer&hl=en&gl=us&ct=clnk&cd=2
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PHOTOHETEROTROPHS
• LIGHT AS ENERGY SOURCE
• NEED ORGANIC COMPOUNDS OR HYDROGEN GAS AS ELECTRON DONOR
• GREEN NONSULFUR BACTERIA
• PURPLE NONSULFUR BACTERIA
• http://www.sinauer.com/perry/MicrobialLife05.pdf
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CHEMOAUTOTROPHS
• REDUCED INORGANIC MOLECULES ACT AS CARBON AND ENERGY SOURCES
• INORGANIC MOLECULE FINAL ELECTRON ACCEPTOR
• CARBON DIOXIDE ACTS AS CARBON SOURCE
• ENERGY OBTAINED FROM H2, NH3, H2S, ELEMENTAL SULFUR, NO2 and Fe+2.
• http://uweb.cas.usf.edu/~kscott/chemoautotrophy.htmhttp://highered.mcgraw-hill.com/sites/0078664276/student_view0/unit1/chapter4/check_challenge_quiz_3.html
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CHEMOHETEROTROPHS
• USE PREFORMED CARBON COMPOUNDS AS ENERGY AND CARBON SOURCE
• USUALLY THE SAME ORGANIC COMPOUND
• FUNGI, ANIMALS, PROTOZOA AND MOST BACTERIA
• http://www.bact.wisc.edu/bact100/origins.html
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SAPROPHYTE VS PARASITE
• SAPROPHYTE USES DEAD ORGANIC MATTER
• PARASITE USES LIVING HOST
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MICROBIAL NUTRITION
• http://www.mansfield.ohio-state.edu/~sabedon//biol2015.htm
• http://www.agen.ufl.edu/~chyn/age4660/lect/lect_02/lect_02.htm
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MACRONUTRIENTS
• NEEDED IN LARGE AMOUNTS• CARBON• HYDROGEN• OXYGEN• NITROGEN• PHOSPHORUS• SULFUR• http://www.biologie.uni-hamburg.de/b-online/
library/micro229/terry/229sp00/lectures/nutrition.html
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CARBON
• FORMS CARBON SKELETON OF ORGANIC MOLECULES
• CARBON DIOXIDE MOST COMMON SOURCE
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OXYGEN
• FOUND IN MOST ORGANIC MOLECULES
• ATMOSPHERIC
• CHEMICALLY BOUND
• OXYGENASES
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TOXIC OXYGEN COMPOUNDS
• OXYGEN ITSELF• HYDROGEN PEROXIDE• SUPEROXIDE RADICAL• HYDROXYL RADICAL• http://www.sigmaaldrich.com/
Area_of_Interest/Biochemicals/Enzyme_Explorer/Cell_Signaling_Enzymes/Superoxide_Dismutase.html
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ENZYMES THAT DETOXIFY FREE RADICALS
• SUPEROXIDE DISMUTASE
• CATALASE• PEROXIDASE
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OXYGEN RELATIONSHIPS
• OBLIGATE AEROBES• FACULATIVE ANAEROBES OR
FACULATIVE AEROBES• MICROAEROPHILES• AEROTOLERANT ANAEROBES• OBLIGATE ANAEROBES• http://www.jlindquist.net/generalmicro/
dfthiognf.html
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HYDROGEN
• FOUND IN ORGANIC MOLECULES
• CHEMIOSOMOSIS
• REDUCES CARBON DIOXIDE IN CALVIN CYCLE
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PHOSPHORUS
• FOUND IN ATP
• PHOSPHOLIPIDS
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NITROGEN
• COMPONENT OF NUCLEIC ACIDS
• COMPONENT OF PROTEINS
• COMPONENT OF COENZYMES
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SULFUR
• COMPONENT OF ACETYL COA
• COMPONENT OF PROTEINS
• MOST MICROBES USE SULFATE
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OTHER ELEMENTS USED BY MICROBES
• POTASSIUM
• MAGNESIUM
• IRON
• CALCIUM
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IRON
• COMPONENT OF HEME GROUPS
• FERRIC AND FERROUS FORMS
• CHELATORS INCREASE ABSORPTION
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MICROELEMENTS
• MOLYBEDNUM
• COBALT
• COPPER
• NICKEL
• MANGANESE
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OTHER ELEMENTS USED BYA RELATIVE FEW ORGANISMS
• SELENIUM & TUNGSTEN– ARCHAEBACTERIA
• SODIUM AND CHLORIDE IONS– HALOPHILES
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ACCESSORY GROWTH FACTORS
• VITAMINS
• FATTY ACIDS
• AMINO ACIDS
• PURINES AND PYRMIDINES
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VITAMINS
• DIFFERENT ORGANISMS HAVE DIFFERENT VITAMIN NEEDS
• NEEDS CAN VARY WITH CONDITIONS
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FATTY ACIDS
• SOME BACTERIA AND PROTOZOA NEED FATTY ACIDS
• MYCOPLASMAS NEED CHOLESTEROL OR OTHER STEROLS
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AMINO ACIDS
• NEEDS VARY
• MUST INCORPORATE INTO MEDIA
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PURINES AND PYRIMIDINES
• SOME MICROBES ARE UNABLE TO SYNTHESIZE THEIR OWN
• MUST BE INCLUDED IN MEDIAADENINE THYMINE
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EUTROPHIC VS OLIGOTROPHIC ENVIRONMENTS
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EUTROPHIC ENVIRONMENTS
• ABUNDANCE OF NUTRIENTS
• NO LIMITING FACTORS
• CULTURAL EUTROPHICATION
• http://www.umanitoba.ca/institutes/fisheries/eutro.html
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SOURCES OF CULTURAL EUTROPHICATION
• MAN’S ACTIVITY INCREASES NUTRIENTS IN ENVIRONMENT– FARMING– SEWAGE
• INCREASE BIOLOGICAL OXYGEN DEMAND– BOD
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OLIGOTROPHIC ENVIRONMENTS
• FEW NUTRIENTS• STREAMS • RIVERS • SOME SOILS
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LABORATORY CULTURE
http://www.bact.wisc.edu/Microtextbook/index.php?
module=Book&func=displayarticle&art_id=26
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NUTRIENT UPTAKE
• SPECIFIC• OFTEN MUST MOVE AGAINST
CONCENTRATION GRADIENT• http://users.rcn.com/jkimball.ma.ultranet/Bi
ologyPages/D/Diffusion.html#Facilitated_diffusion
• http://web.mit.edu/esgbio/www/cb/membranes/transport.html
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TRANSPORTATION MECHANISMS
• DIFFUSION• OSMOSIS• FACILITATED
DIFFUSION• ACTIVE
TRANSPORT• GROUP
TRANSLOCATION
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DIFFUSION
• SMALL NONPOLAR SUBSTANCES
• DOWN CONCENTRATION GRADIENT
• CARBON DIOXIDE• OXYGEN• CARBON DIOXIDE
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• MOLECULES SPREAD FROM AREAS OF HIGH CONCENTRATIION, TO AREAS OF LOW CONCENTRATION
• . MOLECULES EVENTUALLY EVENOUT THROUGHOUT A SPACE - EQUILIBRIUM
• CONCENTRATION GRADIENT - A DIFFERENCE BETWEEN CONCENTRATIONS IN A SPACE
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FACILITATED DIFFUSION
• PASSIVE PROCESS
• PERMEASES
• DOES NOT HAPPEN MUCH IN PROKARYOTES
• MORE IMPORTANT IN EUKARYOTES
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• http://bio.winona.msus.edu/berg/ANIMTNS/FacDiff.htm
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ACTIVE TRANSPORT
• MOVE MATERIALS AGAINST CONCENTRATION GRADIENT
• USES ENERGY TO POWER
• CARRIER MOLECULES
• ANTIPORT
• SYMPORT OR COTRANSPORT
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SYMPORT OR COTRANSPORT
• http://bio.winona.msus.edu/berg/ANIMTNS/symport.htm
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ANTIPORT
• http://bio.winona.msus.edu/berg/ANIMTNS/ANTIport.htm
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• http://www.brookscole.com/chemistry_d/templates/student_resources/shared_resources/animations/ion_pump/ionpump.html
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THE PRICE OF ACTIVE TRANSPORT
• ATP OR OTHER PHOSPHATE MOLECULES
• PROTON MOTIVE FORCE
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GROUP TRANSLOCATION
• MOLECULES ARE MODIFIED AS THEY CROSS CELL MEMBRANE
• PHOSPHOPHENOLPYRUVATE: SUGAR PHOSPHOTRANSFERASE
• WIDESPREAD IN PROKARYOTES• VARIETY OF FORMS
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http://www.cat.cc.md.us/courses/bio141/lecguide/unit1/prostruct/group.html
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ENDOCYTOSIS VS EXOCYTOSIS
EUKARYOTIC MECHANISM
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ENDOCYTOSIS
• MOVEMENT OF MATERIALS INTO THE CELL– PHAGOCYTOSIS = ‘CELL EATING’
• BY MACROPHAGES & WBCS• PARTICLE BINDS TO RECEPTOR PROTEIN• WHOLE BACTERIA OR VIRUSES ARE ENGULFED & LATER
DIGESTED
– PINOCYTOSIS = CELL DRINKING• NO RECEPTOR PROTEINS
– RECEPTOR-MEDIATED ENDOCYTOSIS = SELECTIVE INPUT
• MECHANISM BY WHICH HIV VIRUS ENTERS CELLS
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Pinocytosis and Phagocytosis
• Pseudopods extend to form phagosome
• Lysosome joins it
• No pseudopods form• Nonselective drinking of extracellular
fluid
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PINOCYTOSIS
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PHAGOCYTOSIS
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Receptor-Mediated Endocytosis• Mechanism for uptake of specific
substances -- ligands
• Desired substance binds to receptor protein in clathrin-coated pit region of cell membrane causing membrane to fold inward
• Vesicles become uncoated & combine with endosome
• Receptor proteins separate from ligands and return to surface
• Ligands are digested by lysosomal enzymes or transported across cell -- epithelial cell crossing accomplished
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• http://cellbio.utmb.edu/cellbio/recend.htm#Menu
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EXOCYTOSIS
• MOVEMENT OF MATERIALS OUT OF CELL
• ‘CELL VOMITING’• Vesicles form inside cell,
fuse to cell membrane
• Release their contents– digestive enzymes,
hormones, neurotransmitters or waste products
• replace cell membrane lost by endocytosis
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http://bio.winona.msus.edu/berg/ANIMTNS/Secrtion.htm
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IRON UPTAKE
• USED FOR CYTOCHROMES AND ENZYMES
• FERRIC IRON INSOLUBLE
• SIDEROPHORES
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MEDIA USED IN LABORATORY CULTURE
http://www.life.umd.edu/classroom/bsci424/BSCI223WebSiteFiles/Chapter6.htm
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CLASSIFICATION OF MEDIA
• PHYSICAL FORM
• CHEMICAL CHARACTERISTICS
• FUNCTIONAL TYPES
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LIQUID MEDIA
• AQUEOUS FORMULATION• DOES NOT GEL OR
SOLIDIFY AT ROOM TEMP• FLOW FREELY• BROTHS• MILKS• NUTRIENT SOLUTIONS
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SEMISOLID MEDIA
• SOLID AT ROOM TEMP• GELATINOUS TEXTURE• 0.3% TO 0.5% AGAR• USED TO RESTRICT
MOVEMENT OF MOTILE MICROBES, GROW MICROAEROPHILES OR ANAEROBES
• SIM AGAR• MOTILITY MEDIA
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SOLID MEDIA
• PROVIDES FIRM SURFACE
• LIQUEFIABLE OR NONLIQUEFIABLE
• DISTINCT COLONIES FORM
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LIQUEFIABLE SOLID MEDIA
• REVERSIBLE SOLID MEDIA
• THERMOPLASTIC SOLIDIFYING AGENT
• AGAR OR GELATIN
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GELATIN WAS FIRST SOLIDIFYING AGENT
• DRAWBACKS• CAN BE LIQUID AT
ROOM TEMP• MANY MICROBES
DIGEST
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BENEFITS OF AGAR-AGAR
• COMPLEX POLYSACCHARIDE
• SOLID AT ROOM TEMPERATURE
• LIQUEFIES AT 100 DEGREES C
• DOES NOT RESOLIDIFY UNTIL IT COOLS TO 42 DEGREES C
• CAN BE INOCULATED AND POURED IN LIQUID FORM
• MOLDABLE AND FLEXIBLE FRAMEWORK FOR MOISTURE AND NUTRIENTS
• NOT EASILY DIGESTED
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NONLIQUEFIABLE SOLID MEDIA
• NOT THERMOPLASTIC
• RICE• POTATOE• MEAT• EGG MEDIA• SERUM MEDIA• LESS VERSATILE
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CHEMICAL CONTENT OF MEDIA
• CHEMICALLY DEFINED OR SYNTHETIC
• NONSYNTHETIC OR EMPIRICAL MEDIA
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FUNCTIONAL CATEGORIES
• GENERAL PURPOSE
• ENRICHED
• SELECTIVE
• ENRICHMENT MEDIA
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GENERAL PURPOSE MEDIA
• GROW A BROAD SPECTRUM
• USUALLY NONSYNTHETIC
• NUTRIENT BROTH AND AGAR
• TRYPTIC SOY BROTH AND AGAR
• BLOOD AGAR
• BRAIN-HEART INFUSION AGAR
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ENRICHED MEDIA
• SIMILAR TO GENERAL PURPOSE MEDIA
• ENRICHED WITH BLOOD, SERUM, HEMOGLOBIN, OR GROWTH FACTORS
• BLOOD AGAR
• THAYER MARTIN AGAR
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SELECTIVE METHODS
• SELECTIVE ENRICHMENT
• SELECTIVE REPRESSION
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SELECTIVE ENRICHMENT
• VARYING TEMPERATURE
• pH
• CHEMICAL ENVIRONMENT
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ENRICHMENT MEDIA
• EXTREMELY SELECTIVE MEDIA
• FAVORS GROWTH OF PATHOGENS FOUND IN LOW NUMBERS
• SELENITE
• BRILLIANT GREEN
• POTASSIUM TELLURITE
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SELECTIVE MEDIA
• HAS AGENT/S THAT INHIBITS GROWTH OF CERTAIN MICROBES
• SELECTS FOR GROWTH OF OTHERS
• IMPORTANT FOR IDENTIFICATION FROM SAMPLES
• MANNITOL SALT AGAR
• MacCONKEY AGAR
• EMB AGAR
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SELECTIVE REPRESSION
• STOPS OR SLOWS THE THE GROWTH OF INTERFERING ORGANISMS
• TOXIC CHEMICALS
• TEMPERATURE
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DIFFERENTIAL MEDIA
• EXPLOITS PHYSIOLOGICAL & OTHER CHARACTERISTICS TO DISTINGUISH COLONIES
• CONTAIN REAGENTS
• DO NOT NECESSARILY ENRICH OR REPRESS
• BLOOD AGAR
• EMB
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DIFFERENTIAL MEDIA
• GROW SEVERAL TYPES OF MICROBES
• WILL HAVE VARYING APPEARANCES DUE TO AGENTS
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EXAMPLES OF AGARS
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EMB AGAR
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MacCONKEY AGAR
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MANNITOL SALT AGAR
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ISOLATING PURE CULTURE
KOCH AND HIS PROCEDURES
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KOCH’S POSTULATES
• ORGANISM SHOULD ALWAYS BE PRESENT IN ANIMALS WITH DISEASE
• NOT PRESENT IN ANIMALS THAT ARE HEALTHY
• MUST CULTIVATE ORGANISMS IN PURE CULTURE
• WHEN CULTURE INTRODUCED INTO HEALTHY ANIMAL SHOULD CAUSE DISEASE
• MUST BE REISOLATED FROM EXPERIMENTAL ANIMAL
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PURE CULTURE TECHNIQUE
• SUCCESS DEPENDS ON HAVING ONLY ONE MICROBE IN CULTURE
• SOLID MEDIA ALLOWS FOR DEVELOPMENT OF SINGLE COLONIES
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PETRI PLATES
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STREAK PLATE
• DEVELOPED BY KOCH
• ASEPTIC TECHNIQUE
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POUR PLATE METHOD
• SERIAL DILUTIONS INTO LIQUID AGAR
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PRESERVATION OF MICROBES
• SUBCULTURING
• REFRIGERATION
• DESSICATION
• LYOPHILIZATION
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ENVIRONMENTAL FACTORS AND GROWTH
EFFECTS OF TEMPERATURE
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CARDINAL TEMPERATURES
• MINIMUM TEMPERATURE
• MAXIMUM TEMPERATURE
• OPTIMAL TEMPERATURE
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MAXIMUM TEMPERATURE
• REFLECTS DENATURING OF PROTEINS AND ENZYMES
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MINIMUM TEMPERATURE
• MAYBE DUE TO LOSS OF FLUIDITY IN CELL MEMBRANE
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TEMPERATURE
• EFFECTS ALL RELATIONSHIPS IN CELL
• PSYCHROPHILES• MESOPHILES• THERMOPHILES
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PSYCHROPHILES• BEST AT 15
DEGREES C OR LOWER
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PSYCHROTROPHS
• ARE ALSO MESOTROPHS BEST GROWTH AT 20-30 DEGREES C– CAN GROW SLOWLY
AT LOW TEMPERATURES
– TOLERATE RATHERE THAN BENEFIT FROM THEM
• SPOIL FOOD IN FRIDGE– LISTERIA
MONOCYTOGENES
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MESOPHILES
• MOST MICROBES ARE MESOPHILES
• HUMAN PATHOGENS ARE
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THERMOPHILES
• HEAT LOVERS• ABOVE 55
DEGREES C• CAN GROW IN
BOILING WATER 100 DEGREE C
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HYPERTHERMOPHILES
• GROW AT 90 DEGREES C OR HIGHER
• SOME HAVE MAXIMA ABOVE 100 DEGREES C
• DO NOT GROW WELL BELOW 55
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ENVIRONMENTAL FACTORS AND GROWTH
ACIDITY AND ALKALINITY
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PH AND MICROBIAL GROWTH
• MOST ORGANIMS HAVE PH RANGE
• MOST FALL WITHIN PH 5 - 9
• ACIDOPHILES• NEUTROPHILES• ALAKALOPHILES
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ACIDOPHILES
• LOW PH OPTIMA
• FUNGI TEND TO BE MORE ACID TOLERANT THAN BACTERIA
• OBLIGATE ACIDOPHILIC BACTERIA THIOBACILLUS SULFOLOBUS THERMOPLASMA
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IMPORTANCE OF HYDROGEN IONS TO CELL
MEMBRANE STABILITY• IN NEUTRAL PH CELL MEMBRANE
DISINTEGRATES
• CELL LYSES
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ALKALIPHILES
• HIGH PH OPTIMA FOR GROWTH
• SODA LAKES & HIGH CARBONATE SOILS
• SOME ARE ALSO HALOPHILES
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NEUTROPHILES
• MAJORITY OF MICROBES
• PH 6 - 8
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HOW TO LIVE IN HIGH AND LOW PH ENVIRONMENTS
• INSIDE OF CELL MUST BE AT PH NEAR 7
• MEMBRANE MAY BE IMPERMEABLE TO H+ IONS
• MAY EXCHANGE IONS ACROSS MEMBRANE
• CELL WILL MOVE H+ ACROSS MEMBRANE TO KEEP PH STABLE
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MAINTAINING PH IN THE LAB
• BUFFERS
• WORK OVER NARROW PH RANGE
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ENVIRONMENTAL FACTORS AND GROWTH
WATER AVAILABILTY
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OSMOSIS
• HIGH CONCENTRATION TO LOW CONCENTRATION
• ACROSS CELL MEMBRANE
• http://www.colorado.edu/eeb/web_resources/osmosis/
• http://zoology.okstate.edu/zoo_lrc/biol1114/tutorials/Flash/Osmosis_Animation.htm
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POSITIVE WATER BALANCE
• WATER USUALLY DIFFUSES INTO CELLS– CAUSING LYSIS OR TURGOR PRESSURE
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NEGATIVE WATER BALANCE
• WATER WILL MOVE OUT OF THE CELL
• CAUSING CRENATION OR PLASMOLYSIS
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OSMOTIC EFFECTS IN HIGH SALT ENVIRNOMENTS
• SEA WATER --3% NA CL
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HALOPHILES
• MILD HALOPHILES 1-6%
• MODERATE HALOPHILES 6-15%
• EXTREME HALOPHILES 15-30%
• HALOTOLERANT CAN TOLERATE BUT GROW BEST WITHOUT
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EFFECTS OF HIGH SUGAR ENVIRONMENTS
• OSMOPHILES
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EFFECTS OF LOW WATER (DRY) ENVIRONMENTS
• XEROPHILES
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HOW DO MICROBES GROW IN AREAS WITH LOW WATER
• COMPATIBLE SOLUTES
• PROLINE
• BETAINE
• GLYCEROL
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ENVIRONMENTAL FACTORS AND GROWTH
OXYGEN
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TYPES OF MICROBES
• AEROBES
• MICROAEROPHILES
• FACULATIVE ANAEROBES OR AEROBES
• AEROTOLERANT ANAEROBES
• OBLIGATE ANAEROBES
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TOXIC FORMS OF OXYGEN
• SINGLET OXYGEN 1O2
• SUPEROXIDE ANION 1O2-
• HYDROGEN PEROXIDE H2O2
• HYDROXYL RADICAL ·OH
• ALL ARE BYPRODUCTS OF RESPIRATION
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SINGLET OXYGEN
• NORMAL MOLECULAR OXYGEN
• AT HIGHER ENERGY STATE• EXTREMELY REACTIVE
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SUPEROXIDE ANION
• O2-
• HIGHLY REACTIVE• OXIDIZES ANY ORGANIC MOLECULE
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PEROXIDES
• CAN DAMAGE CELL COMPONENTS• NOT AS TOXIC AS OTHERS
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HYDROXYL RADICAL
• OXIDIZES ORGANIC SUBSTANCES• TRANSIENT• NOT A LOT MADE UNLESS EXPOSED TO IONIZING
RADIATION• SMALL AMOUNTS CAN BE MADE FROM
HYDROGEN PEROXIDE
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ENZYMES THAT DESTROY TOXIC OXYGEN PRODUCTS
• CATALASE
• PEROXIDASE
• SUPEROXIDE DISMUTASE
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ANAEROBIC MICROBES
• FREQUENTLY LACK MEANS TO DETOXIFY OXYGEN BY PRODUCTS
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ENVIRONMENTAL FACTORS AND GROWTH
PRESSURE
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PRESSURE RELATIONSHIPS
• MOST ORGANISMS LIVE AT 1 ATMOSPHERE PRESSURE
• BAROTLERANT• BAROPHILIC
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MICROBIALGROWTH
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MICROBIAL GROWTH
GROWTH OF POPULATIONS NOT INDIVIDUALS
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DOUBLING TIME
• GENERATION TIME
• DEPENDS ON SPECIES
• DEPENDS ON GROWTH CHARACTERISTICS
• TELLS HOW FAST POPULATION IS GROWING
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THE WAY WE GROW
• ENLARGE AND DIVIDE WHEN DOUBLED IN SIZE
• BINARY FISSION
• BUDDING
• BINARY FISSION AND MITOSIS
• MITOSIS AND CYTOKINESIS
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EUKARYOTIC CELL DIVISION
• GO PHASE
• G1 PHASE
• S PHASE
• G2 PHASE
• M PHASEC PHASE
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GO PHASE
• PRIMARY GROWTH PHASE OF CELL
• ENLARGEMENT
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G1 PHASE
• CELL PREPARES FOR DNA REPLICATION
• PRODUCES ENZYMES
• PRODUCES NUCLEOTIDES
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S PHASE
• SYNTHESIS PHASE• REPLICATION OF
GENOME OCCURS
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G2 PHASE
• PRODUCES ORGANELLES FOR M PHASE
• STOCKPILES VITAL CELLULAR PRODUCTS
• CHROMOSOME BEGIN CONDENSING
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M PHASE
• MITOSIS PHASE
• DIVIDES THE NUCLEUS INTO TWO DAUGHTER NUCLEI
• PROPHASE
• METAPHASE
• ANAPHASE
• TELOPHASE
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PROPHASE
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METAPHASE
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ANAPHASE
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TELOPHASE
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C PHASE
• CYTOKINESIS PHASE
• DIVISION OF CYTOPLASM AND ORGANELLES INTO TWO DAUGHTER CELLS
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CYTOKINESIS
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BACTERIAL CELL DIVISION
BUDDING
BINARY FISSION
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BUDDING
• YEAST LIKE PROCESS
• But no mitosis occurs
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BINARY FISSION
• CHROMOSOMES DO NOT SHUT DOWN DURING DIVISION
• CELL MEMBRANE REPLACES MITOTIC SPINDLE
• PRODUCES TWO NEARLY EQUAL SIZED CELLS
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http://www.emc.maricopa.edu/faculty/farabee/BIOBK/BioBookDiversity_2.html
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PROCESSES INVOLVED
• CELL ELONGATION
• DNA REPLICATION
• CELL DIVISION
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CELL ELONGATION
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GRAM NEGATIVE BACTERIA
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GRAM POSITIVE BACTERIA
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DNA REPLICATION
• TIME TABLE CAN VARY
• MAY HAVE MULTIPLE REPLICATION FORKS
• MAY LEAD TO MULTIPLE CHROMOSOMES
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CELL DIVISION
• USUALLY BEGINS AS INVAGINATION
• USUALLY AUTOLYSIS TO FORM TWO CELLS
• GET VARIETY OF ARRANGEMENTS
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GRAM NEGATIVE BACTERIA
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PLANES OF CELL DIVISION
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VIABLE NONCULTURABLE CELLS
• HAVE ACTIVE METABOLISM
• CAN’T BE GROWN ON CONVENTIAL MEDIA
• VIBRIO CHOLERAE, LEGIONELLA PNEUMONIAE
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EXPONENTIAL GROWTH
• POPULATION NUMBERS INCREASE BY A FACTOR OF TWO
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TYPES OF GROWTH
SYNCHRONOUS VS ASYNCHRONOUS
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GROWTH PHASES
• LAG PHASE• LOG PHASE• STATIONARY
PHASE• DEATH PHASE
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HOW DO WE KNOW A CELL IS DEAD
• CANNOT CONTINUE CELLULAR REPAIR
• CANNOT REINITIATE GROWTH WHEN INTRODUCED TO NEW MEDIA
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LABORATORY CULTURE METHODS
BATCH VS CONTINUOUS CULTURE
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BATCH CULTURE
• USUALLY TYPE OF BACTERIAL GROWTH
• SHOWS NORMAL GROWTH CURVE
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CONTINUOUS CULTURE
• CHEMOSTAT
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COLONY GROWTH
• FORMS FROM SINGLE CELL
• DIFFERENT PHASES OF GROWTH IN DIFFERENT PARTS OF COLONY
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MEASURING MICROBIAL GROWTH
DIRECT VS INDIRECT
VIABLE VS TOTAL
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INDIRECT MEASUREMENTS
• MEASURE PROPERTIES OF POPULATION
• TURBIDITY
• DRY WEIGHT
• METABOLIC ACTIVITY
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DIRECT MEASUREMENTS
• DIRECT MICROSCOPIC COUNT
• ELECTRONIC COUNT
• PLATE COUNT
• MOST PROBABLE NUMBER
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VIABLE COUNT
• COUNTS LIVING CELLS
• PLATE COUNTS
• MOST PROBABLE NUMBER
• METABOLIC ACTIVITY
• FILTRATION
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TOTAL COUNT
• TURBIDITY
• DRY WEIGHT
• DIRECT MICROSCOPIC COUNT
• DIRECT ELECTRONIC COUNT
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TURBIDITY
• NUMBER IS PROPORTIONAL TO WEIGHT OF A SAMPLE
• SPECTROPHOTOMETER
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USE OF THE SPECTROPHOTOMETER
• ESTIMATES MASS OF DENSE CULTURES
• CHARTS GROWTH COMPARED TO STANDARD GROWTH CURVE
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• ADVANTAGES|
RAPIDITY
REPRODUCIBLE
• DISADAVANTAGES
CAN BE USED ONLY ON DENSE CULTURES
DOES NOT DISTINGUISH BETWEEN LIVING AND DEAD CELLS
CAN NOT BE USED ON CELLS THAT AGGREGATE
NEED STANDARD CURVE
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DRY WEIGHT
• CENTRIFUGATION OR FILTRATION
• DRYING IN OVEN AT 105 DEGREES C FOR 24 HOURS
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• ADVANTAGES
USED TO MAKE STANDARD CURVE FOR MEASURING CELL MASS
ACCURATE
REPRODUCIBLE
DISADVANTAGES
TIME CONSUMING
TEDIOUS
SAMPLE MUST CONTAIN MORETHAN 10 MILLION CELLS
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METABOLIC ACTIVITY
• RATE OF METABOLITE PRODUCTION
• UTILIZATION OF SUBSTRATE
• REDUCTION OF DYES
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ADVANTAGES
CAN BE USED WITH COMPLEX MEDIA SUCH AS MILK OR SOIL
NO INSTRUMENTS REQUIRED
DISADVANTAGES
INDIRECT MEASURMENT
DOES NOT GIVE AN ACCURATE MEASUREMENT
TIME CONSUMING
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DIRECT COUNT
• PETROFF HAUSER COUNTING CHAMBER
• COULTER COUNTER
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PETROFF HAUSSER COUNTING CHAMBER
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COULTER COUNTER
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MICROSCOPIC VS ELECTRONIC COUNTS
• USED IN DIFFERENT SITUATIONS
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MICROSCOPIC COUNTS
• ADVANTAGES– NO EXPENSIVE
EQUIPMENT
– ONLY WAY TO COUNT IF SAMPLE CONTAINS FOREIGN MATERIALS
• SLOW• TEDIOUS• NOT USEFUL FOR
DILUTE CULTURES
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ELECTRONIC COUNT
• ADVANTAGES– RAPID
– ACCURATE IF CELLS ONLY ARE PRESENT
• EXPENSIVE• CAN’T USE IF
FOREIGN PARTICLES ARE PRESENT
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PLATE COUNT
• POUR PLATE
• SERIAL DILUTIONS
• COUNT PLATES BETWEEN 3O AND 300 COLONIES
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• ADVANTAGES– EXTREMELY
SENSITIVE
– DOES NOT NEED COMPLICATED EQUIPMENT
• DISADVANTAGES– SAMPLING ERRORS
– TIME CONSUMING
– TEDIOUS
– LARGE NUMBERS MUST BE COUNTED TO REDUCE SAMPLING ERROR
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MOST PROBABLE NUMBER
• A SINGLE LIVE CELL CAN GIVE RISE TO A TURBID CULTURE
• SERIAL DILUTIONS• USES STATISTICAL
TABLE
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• ADVANTAGES– ALLOW COUNTS OF
MICROBES THAT ARE DIFFICULT TO GROW ON SOLID MEDIA
– CAN BE USED TO COUNT CELLS IN MIXED LIQUID CULTURE
• DISADVANTAGE– TIME CONSUMING
– TEDIOUS
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FILTRATION
• PREPARATION FOR OTHER METHODS
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MICROBIAL DEATH
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TERMS
• STERILIZATION
• DISINFECTION
• SANITATION
• DECONTAMINATION
• ANTISEPSIS
• MICROBIOSTATIC
• MICROBIOCIDAL
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RATE OF MICROBIAL DEATH
• EXPONENTIAL DEATH
• ASYNCHRONOUS
• CAN BE CALCULATED
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DECIMAL REDUCTION TIME
• D VALUE
• TIME IT TAKES TO KILL 90 PERCENT OF POPULATION
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FACTORS THAT AFFECT D VALUE
• TEMPERATURE
• TYPE OF MICROBE
• PHASE OF GROWTH
• PRESENCE OF OTHER SUBSTANCES
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PROBABILITY OF STERIZATION
• 90 PERCENT OF POPULATION DIES
• EVENTUALLY WE HAVE THE PROBABILITY OF NO MICROBES IN THE POPULATION
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DESIGNING A STERILIZATION PROGRAM
• D VALUE OF TREATMENT
• NUMBER OF CELLS PRESENT
• DEGREE OF CERTAINTY OF STERILITY DESIRED
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THERMAL DEATH POINT
• TDP
• LOWEST TEMPERATURE NEEDED TO KILL ALL MICROBES IN A LIQUID SUSPENSION IN 10 MINUTES
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THERMAL DEATH TIME
• TDT
• MINIMAL TIME TO KILL ALL MICROBES IN A LIQUID SUSPENSION AT A GIVEN TEMPERATURE
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PHYSICAL CONTROLS OF MICROBIAL GROWTH
• HEAT
• COLD
• RADIATION
• FILTRATION
• DRYING
• OSMOTIC STRENGTH
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HEAT
• INEXPENSIVE
• EFFECTIVE
• DRY HEAT
• MOIST HEAT
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AUTOCLAVE
• USES STEAM TO STERILZIE
• MAINTAINS PRESSURE AT 103 kPa (15 PSI)
• KILLS ALL BACTERIA, VIRUSES, FUNGI AND ENDOSPORES– EXCEPT STRAIN 121 AND PRIONS– DOES NOT DESTROY ENDOTOXINS
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PASTEURIZATION
• DISINFECTION PROCESS
• MILK, WINES, BEERS……
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COLD
• BY ITSELF DOES NOT KILL
• COLD SHOCK ONLY EXCEPTION
• MICROBIOSTATIC
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FREEZING
• DOES KILL MOST BACTERIA
• DOES KILL EUKARYOTES
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RADIATION
• ELECTROMAGNETIC RADIATION
• ULTRAVIOLET RADIATION
• IONIZING RADIATION
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UV RADIATION
• WAVELENGTH OF 10 TO 400 nm
• 265 nm UV LIGHT MOST LETHAL
• GERMICIDAL LIGHTS
• 253.7 nm LIGHT
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IONIZING RADIATION
• X-RAYS• GAMMA RAYS• CAUSE CHAIN OF
IONIZATIONS• KILL CELLS
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FILTRATION
• MICROBES EXCEPT VIRUSES REMOVED
• NOT STERIZATION
• USED FOR HEAT
• LABILE MEDIA
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DRYING
• EVAPORATION
• SUBLIMATION
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EVAPORATION
• REMOVAL OF WATER
• USED FOR FOOD
• SELDOM USED IN
• MICRO LAB
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LYOPHILIZATION
• SUBLIMATION• DIRECT
CONVERSION FORM SOLID STATE TO GASEOUS STATE
• VACUUM
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OSMOTIC STRENGTH
• USED TO PRESERVE FOOD
• SUGAR• SALT
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CHEMICAL CONTROL OF MICROBES
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TERMS
• CHEMOTHERAPEUTIC AGENTS
• GERMICIDES
• GERMISTATS
• DISINFECTANTS
• ANTISEPTICS
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SELECTING A GERMICIDE
• HIGH ACTIVITY GERMICIDE
• INTERMEDIATE ACTIVITY GERMICIDE
• LOW ACTIVITY GERMICIDE
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SELECTING A GERMICIDE
• HIGH ACTIVITY GERMICIDE
• INTERMEDIATE ACTIVITY GERMICIDE
• LOW ACTIVITY GERMICIDE
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TESTING GERMICIDES
• SERIAL DILUTIONS
• PHENOL COEFFICIENT
• PAPER DISC METHOD
• USE DILUTION TEST
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TYPES OF GERMICIDES
• PHENOLS , PHENOLICS & BISPHENOLS
• BIGUANIDES
• ALCOHOLS
• HALOGENS AND PEROXYGENS
• HEAVY METALS
• SURFACTANTS
• ALKYLATING AGENTS
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PHENOLS, PHENOLICS & BISPHENOLS
• PHENOL– CARBOLIC ACID
• PHENYLPHENOL• HEXACHLOROPHENE• INACTIVATE VITAL CELLULAR
PROTEINS• PHENOLICS DISRUPT CELL
MEMBRANES
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BIGUANIDES
• CHLORHEXIDINE
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ALCOHOLS
• DISRUPT LIPIDS IN CELL MEMBRANES
• DISRUPT PROTEINS• ETHANOL• ISOPROPANOL
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HALOGENS
• OXIDIZING AGENTS• INACTIVATE ENZYMES• ATTACK SULFHYDRYL
GROUPS• IODINE• CHLORINE
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PEROXYGENS
OZONE, HYDROGEN PEROXIDE AND PERACETIC ACID
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OZONE
• HIGHLY REACTIVE• SUPPLEMENTS
CHLORINE IN WATER TREATMENT
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HYDROGEN PEROXIDE
• OXIDIZING AGENT• WORKS LIKE
HALOGENS• 3% AS ANTISEPTIC
OR DISINFECTANT
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PERACETIC ACID
• EFFECTIVE AGAINST SPORES AND VIRUSES
• FOOD AND MEDICAL INSTRUMENT USES
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HEAVY METALS
• MERCURY
• SILVER
• MERCURIC CHLORIDE
• MERTHIOLATE AND MERCUROCHROME
• COLLOIDAL SILVER AND SILVER SALTS
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SURFACTANTS
• HYROPHILIC AND HYDROPHOBIC PORTIONS
• FORM EMULSIONS
• QUATERNARY AMMONIUM SALTS
• ANIONIC SURFACTANTS
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ALKYLATING AGENTS
• FORMALDEHYDE--FORMALIN
• ETHYLENE OXIDE
• GLUTARALDEHYDE
• ATTACH SHORT CARBON CHAINS INTO ENZYMES
• INACTIVATES THEM AND CELL DIES
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FORMALDEHYDE
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GLUTARALDEHYDE
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ETHYLENE OXIDE
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Microbial Characteristics and Microbial Control
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CHEMOTHERAPEUTIC AGENTS
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INTERNET RESOURCES
• http://gsbs.utmb.edu/microbook/ch011.htm
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ANTIMICROBIAL CHEMOTHERAPY
THE USE OF CHEMICALS TO CONTROL OR PREVENT
INFECTION
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CHEMOTHERAPY
THE USE OF CHEMICALS TO TREAT, RELIEVE OR PREVENT
DISEASE
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SOURCES OF ANTIBACTERIAL AGENTS
• http://helios.bto.ed.ac.uk/bto/microbes/penicill.htm
• BACTERIA– STREPTOMYCES– BACILLUS
• FUNGI– PENICILLIUM– CEPHALOSPORIUM
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SCOPE OF ANTIMICROBIAL ACTION
• NARROW SPECTRUM
• BROAD SPECTRUM
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INTERACTIONS BETWEEN HOST, MICROBE, AND DRUG• DRUG ADMINISTRATION
• DRUG ABSORPTION AND DISTRIBUTION
• DESTRUCTION OR INHIBITION OF MICROBE
• DRUG INACTIVATION BY HOST
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DRUG ADMINISTRATION
• PER OS
• INTRAMUSCULAR INJECTION
• INTRAVENOUSLY
• TOPICAL
• INJECTION INTO BODY CAVITY OR SUBCUTANEOUSLY
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INTERACTIONS BETWEEN HOST, MICROBE, AND DRUG• DRUG ADMINISTRATION
• DRUG ABSORPTION AND DISTRIBUTION
• DESTRUCTION OR INHIBITION OF MICROBE
• DRUG INACTIVATION BY HOST
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DRUG ELIMINATION
• SOME DRUGS METABOLIZED IN LIVER
• MOST ELIMINATED BY KIDNEYS
• PROBENECID SLOWS RATE OF EXCRETION BY KIDNEYS
• SOME DRUGS EXCRETED BY LIVER IN BILE AND IN FECES
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MECHANISMS OF DRUG ACTION
INTERACTION BETWEEN DRUG AND MICROBE
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SELECTIVE TOXICITY
• INHIBIT OR KILL MICROBES
• HAVE LITTLE IF ANY EFFECT ON HOST TISSUES
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MECHANISMS OF DRUG ACTION
• INHIBIT CELL WALL SYNTHESIS
• INHIBIT NUCLEIC ACID SYNTHESIS
• INTERFERE WITH PROTEIN SYNTHESIS
• INTERFERE WITH THE FUNCTION OF THE CELL MEMBRANE
• ANTIMETABOLITE
• INACTIVATE ENZYMES
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ANTIMICROBIAL DRUGS THAT AFFECT THE BACTERIAL CELL
WALL
• PEPTIDOGLYCAN TARGET
• CELL WILL LYSE WITHOUT IT
• PENICILLINS
• CEPHALOSPORINS
• VANCOMYCIN
• BACITRACIN
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PENICILLIN
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CEPHALOSPORIN
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VANCOMYCIN
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BACITRACIN
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ISONIAZID
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ETHAMBUTOL
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DRUGS THAT DISRUPT THE CELL MEMBRANE
• DIE FROM METABOLIC INSUFFICIENCY
• LYSIS
• POLYMIXINS
• POLYENES
• IMIDAZOLES
• ACTS ON LIPIDS IN MEMBRANES
• NOT AS GOOD SELECTIVE TOXICITY
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POLYMIXINS
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NYSTATIN
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AMPHOTERCIN B
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IMIDAZOLES
. .
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DRUGS THAT INHIBIT PROTEIN SYNTHESIS
• AFFECT 70s RIBOSOME
• 30s SUBUNIT----ie. AMINOGLYCOSIDES
TETRACYCLINE
• 50s SUBUNIT----ie. CHLORAMPHENICOL
ERYTHROMYCIN
• BACTERICIDAL--AMINOGLYCOSIDES
• BACTERIOSTATIC--ALL OTHER GROUPS
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STREPTOMYCIN
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ERYTHROMYCIN
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TETRACYCLINES
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CHLORAMPHENICOL
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DRUGS THAT INHIBIT NUCLEIC ACID SYNTHESIS
• LONG METABOLIC PATHWAY
• TOPOISOMERASES--QUINOLONES
• POLYMERASES—RIFAMPIN
• FLUCYTOSINE
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RIFAMIPIN
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QUNINOLONES
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FLUCYTOSINE
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DRUGS THAT ARE ANTIMETABOLITES
• ANTIMETABOLITES• INHIBITION OF
FOLIC ACID SYNTHESIS
• SULFONAMIDES• FLUCYTOSINE
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ANTIVIRAL DRUGS
• ACYCLOVIR
• AMANTIDINE
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DRUGS THAT INACTIVATE ENZYMES
• PENICILLIN & CEPHALOSPORINS
• VANCOMYCIN
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EXAMPLES OF SIDEFFECTS
• TOXICITY
• HYPERSENSITIVITIES
• DISRUPTION OF NORMAL FLORA
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TOXICITY
SOME DRUGS HAVE TOXIC EFFECTS ON PATIENTS
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HYPERSENSITIVITIES
• SOME ALLERGENS
• SOME HAPTENS
• MILD TO SERVERE SKIN RASHES
• ANAPHYLACTIC SHOCK
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DISRUPTION OF NORMAL FLORA
• ESPECIALLY BROAD SPECTRUM
• SKIN, DIGESTIVE, UPPER RESPIRATORY, UROGENITAL TRACT
• CANDIDA
• SUPERINFECTION
• LACTINEX IS GIVEN TO COUNTERACT
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DRUG RESISTANCE
• ADAPTIVE RESPONSE OF MICROORGANIMS
• RESULT OF DRUG THERAPY, GENETIC VARIABILITY AND NATURAL SELECTION
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REASONS FOR SUPERINFECTIONS
• OFTEN PATIENTS ARE DEBILITATED AND HAVE LESS RESISTANCE
• THE CARE ENVIRONMENT OFTEN HAS DRUG RESISTANT ORGANISMS PRESENT
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DRUG RESISTANCE• GENETIC MECHANISMS• NONGENETIC MECHANISMS• http://images.google.com/imgres?imgurl=http://
microvet.arizona.edu/Courses/MIC438/decker/AntibioticRes/20-T02_Activity.jpg&imgrefurl=http://microvet.arizona.edu/Courses/MIC438/decker/AntibioticRes/AntibioticResistance.html&h=361&w=800&sz=55&tbnid=GLwmlTgTlJqRoM:&tbnh=64&tbnw=142&hl=en&start=7&prev=/images%3Fq%3Dspectrum%2Bof%2Bantibiotic%2Baction%26svnum%3D10%26hl%3Den%26lr%3D%26sa%3DG
• http://images.google.com/imgres?imgurl=http://www.wiley.com/college/pratt/0471393878/student/activities/bacterial_drug_resistance/resistance_transfer_web.jpg&imgrefurl=http://www.wiley.com/college/pratt/0471393878/student/activities/bacterial_drug_resistance/&h=285&w=400&sz=27&tbnid=mH0fjM9zuN7zXM:&tbnh=85&tbnw=120&hl=en&start=9&prev=/images%3Fq%3DDRUG%2BRESISTANCE%26svnum%3D10%26hl%3Den%26lr%3D%26sa%3DG
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NONGENETIC MECHANISMS
• EVASION
• L FORMS
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GENETIC MECHANISMS
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DEVELOPMENT OF DRUG RESISTANCE
• MUTATIONS
• TRANSFORMATION
• TRANSDUCTION
• CONJUGATION
• RESISTANCE FACTORS
• http://www.methylgene.com/images/gestion/BetaLactamase.swf
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MUTATIONS
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CONJUGATION
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TRANSFORMATION
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GENERALIZED TRANSDUCTION
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SPECIALIZED TRANSDUCTION
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SPECIFIC MECHANISMS• SYNTHESIS OF ENZYMES THAT INACTIVATE
DRUG• DECREASE IN CELL MEMBRANE
PERMEABILITY AND UPTAKE• CHANGE IN NUMBER OF AFFINITY OF
RECEPTOR SITES• MODIFICATION OF METABOLIC PATHWAYS• DORMANCY• L FORMS
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MECHANISMS OF DRUG INACTIVATION—ENZYMES
• ENZYMES THAT ALTER DRUG STRUCTURE
• BETA-LACTAMASES
• PENICILLINASE
• CEPAHLOSPORINASE
• STAPHYLOCOCCUS AUREUS AND NESSERIA GONORRHOEAE
• SOME GRAM NEGATIVE BACTERIA
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MECHANISMS OF DRUG INACTIVATION--ADDITION OF
FUNCTIONAL GROUPS• AMINOGLYCOSIDES
• CHLORAMPHENICOL
• PREVENTION OF ATTACHMENT TO BACTERIAL RIBOSOME
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DECREASED PERMEABILITY OF CELL TO DRUG
• GRAM NEGATIVE OUTER MEMBRANE
• PUMP THE DRUG BACK OUT OF CELL
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CHANGE DRUG RECEPTORS
• ALTER NATURE OF DRUG’S TARGET
• ALTER PROTEINS --RIFAMPIN AND STREPTOMYCIN
• ALTER 50s RIBOSOME--CLINDAMYCIN, LINCOMYCIN, ERYTHROMYCIN
• CHANGE BINDING SITE IN CELL WALL-PENICILLIN & METHAICILLIN
• DECREASE SYNTHESIS OF ERGOSTEROL IN FUNGAL CELL WALL--AMPHOTERCIN B
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CHANGE METABOLIC PATHWAYS
• DEVELOP ALTERNATE PATHWAY OR ENZYME
• SULFONAMIDE AND TRIMETHROPIM RESISTANCE
• SHUT DOWM PATHWAY
• FLUCYTOSINE
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THE ROLE OF NATURAL SELECTION IN DRUG RESISTANCE
• OCCURS IN WHOLE POPULATION
• VARIATION IN POPULATION
• RESISTANT STRAINS MAY BE IN LOW NUMBERS
• IN PRESENCE OF ANTIBACTERIAL AGENT NON RESISTANT DIE
• RESISTANT FORMS INCREASE DUE TO REPRODUCTION
• http://www.geocities.com/Heartland/7547/antibios.html
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FIRST, SECOND AND THIRD LINE DRUGS
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FIRST, SECOND AND THIRD GENERATION DRUGS
• DRUGS THAT ARE DERIVATIVES OF ONE ANOTHER
• MAY BE USED AS LINE DRUGS
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CROSS RESISTANCE
• RESISTANCE TO TWO OR MORE DRUGS
• BETA-LACTAMASE
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NOSCOMIAL DRUG RESISTANT INFECTIONS
• SULFONAMIDES FIRST ANTIBACTERIAL AGENT
• FIRST TO HAVE RESISTANT SPECIES DEVELOP
• PENICILLIN NEXT ADVANCE
• AT FIRST 95+% OF ALL STAPH. AUREUS WERE SUSCEPTIBLE
• NOW 95+% OF ALL STAPH. AUREUS ARE RESISTANT
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EXAMPLES OF RESISTANT SPECIES
• STAPHYLOCOCCI
• GONOCOCCI
• SALMONELLA
• NEISSERIA
• PSEUDOMONAS
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NOSOCOMIAL INFECTIONS
• SICK PEOPLE FREQUENT HOSPITALS
• TEND TO BE MORE SEVERELY ILL
• HAVE LOWERED RESISTANCE TO DISEASE
• HOSPITALS USE LOTS OF ANTIBACTERIAL AGENTS
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HOW TO REDUCE ANTIBACTERIAL AGENTS THAT ARE RESISTANT NOSOCOMIAL
INFECTIONS• WASH YOUR HANDS
• LIMIT THE USE OF ANTIBACTERIAL AGENTS
• USE SENSITIVITY TESTS
• USE ANTIBACTERIAL AGENTS UNTIL ORGANISM IS COMPLETELY ERADICATED
• USE DRUG COMBINATIONS
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DETERMINING MICROBIAL SENSITIVIES TO ANTIMICROBIAL
AGENTS• DISK DIFFUSION METHOD
• DILUTION METHOD
• SERUM KILLING POWER
• AUTOMATED METHODS
• http://www.lancet.co.za/assets/pdf/news/Antimicrobial_Susceptibility_Tests.pdf
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DISK DIFFUSION METHOD
• KIRBY BAUER METHOD
• ZONES OF INHIBITION
• 24-48 HOURS• SENSITIVE• MODERATELY
SENSITIVE• RESISTANT
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DILUTION METHOD• STANDARDIZED
SHALLOW WELLS• SPECIMEN INTRODUCED
INTO BROTH AND DRUG• 16-20 HOURS• MINIMUM INHIBITORY
CONCENTRATION• MINIMUM
BACTERIOCIDAL CONCENTRATION
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SERUM KILLING POWER
• PATIENT’S SERUM• BACTERIAL
SUSPENSION• LOOK FOR
TURBIDITY
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INCREASING DRUG EFFECTIVENESS
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THE IDEAL CEHMOTHERAPEUTIC AGENT
• SOLUBLE IN BODY FLUIDS
• SELECTIVE TOXICITY
• NOT EASILY ALTERED TO TOXICITY
• NONALLERGENIC
• STABILITY--ABILITY TO MAINTAIN CONSTANT THERAPEUTIC CONCENTRATION
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THE IDEAL CHEMOTHERAPEUTIC AGENT
• FEW ORGANISMS ARE RESISTANT
• LONG SHELF LIFE
• REASONABLE COST