microbial nutrition gary andersen reference chapter 2,5,6, 7, 8, 9 microbiology by tortora, funke...
TRANSCRIPT
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Microbial Nutrition
Gary AndersenReference Chapter 2,5,6, 7, 8, 9 Microbiology by
Tortora, Funke & Case
ChemicalSubstances
Metabolism
Growth
Cell
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What do microbes eat?
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Composition and Nutrition of Cells
• 96% of cells are composed of HCNOPS.• Macronutrients are nutrients required in
larger quantities. (carbohydrates, proteins, fats and other CHO molecules)
• Micronutrients are nutrients required in trace amounts. (Mn, Zn, Cu, Ni)
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Where are the Elements Used?
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Sources of Nutrients
• Carbon – CO2 in air and rocks organisms• Nitrogen – N2 in air, NO2, NO3, NH4 in soil and
water organisms (NH3)• Oxygen – O2 in air, inorganic salts SO4, PO4, NO3,
H2O• Hydrogen – Water, Organic compounds in
organisms• Phosphorus – Rocks and minerals organisms• Sulfur – Rocks and minerals organisms
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Microbe Nutritional Types• Autotrophs – “Self feeder”
– Photoautotrophs – Photosynthetic (energy from light)
– Chemoautotrophs – Energy from simple inorganic chemicals
• Methanogens – Metabolize H2 and CO2 into CH4 and H2O
• Heterotrophs – “Other feeder”– Chemoheterotroph – Obtain carbon and energy from
organic compounds. CnH2nOn + O2 CO2 + H2O + ATP (Adenosine tri-phosphate)
• Saprobe – Free living organisms that feed on dead organisms
• Parasite – Derive nutrients from the tissues of hosts.
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Microbial Clean-Up: The 1989 Exxon Valdez oil spill left great quantities of pooled oil on sites in the Gulf of
Alaska, such as on Green Island
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Microbial Clean-Up: Bioremediation in 1989, by the application of nutrients (nitrogen and phosphorus) to the shoreline accelerated the bacterial biodegradation of the oil into carbon dioxide and water
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Microbial Clean-Up: In 1991, the area was surveyed and found to be mostly cleared of oil, with no further treatment recommended
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How do Microbes Eat?
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Transport Mechanisms• Passive Transport
– Diffusion – Movement of molecules from a high concentration to a low concentration.
– Facilitated Diffusion- diffusion assisted by conformational change in a protein molecule.
– Osmosis – Diffusion of water through a semipermeable membrane
• Active Transport – Moving particles against the diffusion gradient using membrane proteins and expending energy.– Endocytosis – Engulfing with cell membrane and forming a
vacuole.• Phagocytosis – Engulfing of cells or particles by the cell membrane• Pinocytosis – Engulfing of liquids by the cell membrane
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Passive Transport• Osmosis animation:
http://www.tvdsb.on.ca/westmin/science/sbi3a1/Cells/Osmosis.htm
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Comparing Solutions• Hypertonic Solution – Higher solute concentration.
Cells in hypertonic solutions lose water and the cell membrane shrinks away from the cell wall. (Salt on a slug)
• Hypotonic Solution – Lower solute concentration. Cells in hypotonic solutions take on water and swell. (Prune wrinkles of skin)
• Isotonic Solutions – Solutions that have reached an equilibrium with a cell or another solution. The concentration of solute is equal and the diffusion of water proceeds at equal rates.
(See page 93 of text for what happens to a cell in hypertonicand hypotonic solutions.)
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Animations of Passive and Active Transport
• http://programs.northlandcollege.edu/biology/Biology1111/animations/transport1.html
• http://highered.mcgraw-hill.com/olc/dl/120068/bio02.swf
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How do microbes metabolize nutrients?
Fermentation and Respiration
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Enzymes• Provide a surface on which reactions take place
• Active site: the area on the enzyme surface where the enzyme forms a loose association with the substrate
• Substrate: the substance on which the enzyme acts
• Enzyme-substrate complex: formed when the substrate molecule collides with the active site of its enzyme
• Enzymes generally have a high degree of specificity
• Endoenzymes (intracellular)/exoenzymes (extracellular)
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The Action of Enzymes on Substrates to Yield Products
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Each substrate binds to an active site, producing an enzyme-substrate complex. The enzyme helps
a chemical reaction occur, and one or more products are formed
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Competitive Regulation and Inhibition of Enzymes
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Noncompetitive (allosteric) inhibition of enzymes
http://highered.mcgraw-hill.com/olc/dl/120070/bio10.swf
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Factors Influencing Enzymes
• Temperature
• pH
• Concentration of substrate, product, and enzyme
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Relationship between temperature and enzyme activity
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Microbes and Environmental Factors
• Temperature• pH• Oxygen• PressureExtremophiles – Organisms that can survive under
extreme environmental conditions. An interesting source of chemical products.
Interesting Website on Extremophiles and Chemical Products: http://www.mediscover.net/Extremophiles.cfm
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Temperature
Classification
• Psychrophile• Mesophile• Thermophile
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Temperature
• Thermophiles – organisms that grow at >45 degrees C.
• Pyrococcus fumarii is an example of a thermophile that can survive at 113 C.
Thermal pool
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Grand Prismatic Spring
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pH
• Acidophiles – grow at low pH levels. (1-2)
• Alkalinophiles – live at high pH levels. (9-10)
14 Alkaline13121110987 Neutral6543210 Acidic
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Pressure
• Barophiles – organisms that grow at elevated pressure (3-1000 x air pressure). (Found in ocean depths often in thermal vents)
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Presence of Oxygen• Aerobe – Organism able to use O2 in metabolism.• Anaerobe – Organism unable to use O2 in metabolism.
Obligate aerobes - oxygen mandatoryObligate anaerobes - oxygen toxic
Facultative anaerobes – Aerobe that can also live without O2
Microaerophiles - low oxygen levels required.
Aerotolerant - anaerobic metabolism, oxygen not toxic
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Microbial Processing of Oxygen• Step 1: O2
- + O2- + 2H+ H2O2 + O2 (Catalyzing
enzyme is Superoxide dismutase)
• Step 2: H2O2 + H2O2 2H2O + O2 (Catalyzing enzyme is
Catalase)
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Living without Oxygen….Glucose Fermentation Pathways
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Glucose FermentationNet and practical results
• Cells get chemical energy (ATP)
• Fermentation products are natural waste products useful to humans:
1. Fermented beverages
2. Bread
3. Cheese
4. Yogurt
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Using fermentation metabolism to identify microbes: A positive (yellow) mannitol-fermentation test. This test
distinguishes the pathogenic Staphylococcus aureus
(MSA)TestMannitolSaltsAgar
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Comparing Aerobic Respiration with Anaerobic Fermentation
Fermentation• Fermentation yields small amount of ATP (2)• Partial oxidation of carbon atoms (6 C 3 C)
Respiration • Substrate molecules are completely oxidized to
C02 (6 C 1 C)• Far higher yield of ATP (36)• The Krebs Cycle and Electron Transport Chain
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Final Electron Acceptors: Aerobic respiration, anaerobic respiration, and fermentation have
different final electron acceptors
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Microbe Growth
• Binary or transverse fission
• Generation or Doubling Time – the time required for parent cell to form two new daughter cells.
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Microbe Growth
• Lag – new cells require adjustment and enlargement. The cells are not multiplying rapidly.
• Log or exponential – maximum rate of growth
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Microbe Growth 2
• Stationary – death and multiplication balance out. Depleted nutrients and waste buildup.
• Death – limiting factors intensify. May last a long time.
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Calculating Growth of Cells
Nf = (Ni)2nNf = Final population
Ni = Initial population
2n = # cells in generation
n = generation number
Use the table in the handout from the Talaro Appendix A-2 to calculate the number of cells in the generation.
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Measuring growthturbidityplate counts
Measuring Growth Serial Dilutions, Plate Counts and Turbidity
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Calculation of the number of bacteria per milliliter of culture using serial dilution
Pour plate: made by first adding 1.0ml of diluted culture to 9ml of molten agar
Spread plate: made by adding 0.1ml of diluted culture to surface of solid medium
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Counting colonies using a bacterial colony counter
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Bacterial colonies viewed through the magnifying glass against a colony-counting grid
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Which of these plates would be the correct one to count? Why?
Countable number of colonies
(30 to 300 per plate)
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The Petroff-Hausser Counting Chamber
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Turbidity, or a cloudy appearance, is an indicator of bacterial growth in urine in the tube on the left
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A Spectrophotometer: This instrument can be used to measure bacterial growth by determining the degree of light transmission through the culture
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The Streak Plate Method uses agar plates to prepare pure cultures
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A Streak Plate of Serratia marcescens. Note the greatly reduced numbers of growth /colonies in each successive region
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Types of Culture Media
• Natural Media: In nature, many species of microorganisms grow together in oceans, lakes, and soil and on living or dead organic matter
• Synthetic medium: A medium prepared in the laboratory from material of precise or reasonably well-defined composition
• Complex medium: contains reasonably familiar material but varies slightly in chemical composition from batch to batch (e.g. peptone, a product of enzyme digestion of proteins)
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Commonly Used Media
• Yeast Extract
• Casein Hydrolysate
• Serum
• Blood agar
• Chocolate agar
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Selective, Differential, and Enrichment Media
• Selective medium: encourages growth of some organisms but suppresses growth of others (e.g. antibiotics)
• Differential medium: contains a constituent that causes an observable change (e.g. MacConkey agar)
• Enrichment medium: contains special nutrients that allow growth of a particular organism that might not otherwise be present in sufficient numbers to allow it to be isolated and identified
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Three species of Candida can be differentiated in mixed culture when grown on CHROMagar Candida plates
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Identification of urinary tract pathogens with
differential media (CHROMagar)
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Ecological Associations
Mutualism Commensalism Parasitism
Symbiotic
Synergism Antagonism
Non-Symbiotic
Microbial Associations
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Symbiosis (Mutualism)
• Obligatory• Both organisms
benefit.• Examples: algae +
fungus = lichen, termites and trychonympha (a protist)
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Symbiosis (Commensalism)
• One organism benefits and the other is not harmed.
• Examples: Non-pathogenic bacteria on our skin; satellitism between bacteria colonies.
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Symbiosis (Parasitism)
• One organism benefits and the other is harmed.
• Examples: Pathogenic organisms on their host. Plasmodium vivax a protozoan parasite causing malaria.
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Non Symbiotic (Synergism)
• Free living organisms.• Both benefit• The relationship is
optional.• Examples: Shared
metabolism; nitrogen fixing bacteria in the soil and plants
Substance A
Substance B
Substance C
Microorganism 1
Microorganism 2
End Product used by all threemicroorganisms
Microorganism 3
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Non Symbiotic (Antagonism)
• Free living organisms• Organisms compete
for resources.• One organism secretes
a substance toxic to the other.
• Example: Ruminal cellulose digesting bacteria and fungi
GA F05
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End of Microbial Nutrition Slides