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UH / Biology / BIOL 3332 / What is an axenic growth?

What is an axenic growth?

What is an axenic growth?

Description

School: University of Houston
Department: Biology
Course: Elementary Microbiology
Professor: R. knapp
Term: Spring 2016
Tags:
Cost: 25
Name: Chapter 4 Notes
Description: These are the chapter 4 notes that are a combination of notes from the book and lecture
Uploaded: 09/23/2016
8 Pages 193 Views 1 Unlocks
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Chapter 4If you want to learn more check out How to find the equation of a secant line?

Nutrient Supply and Bacterial Growth

  • Essential nutrients cell can’t make itself
  • Must gather from environment
  • Micronutrient required in large amount (mg/L)
  • C, H, O, N, D, S
  • Makeup carbohydrates, lipids, nucleic acids and proteins
  • Mg2+, Fe24, K+, Ca2+
  • Role in enzyme cofactors, aid metabolic process and regulatory signal molecules
  • Micronutrient require small amounts
  • Trace elements
  • Cobalt, copper, manganese, molybdenum, nickel, zinc
  • Enzyme cofactors
  • Not added to growth medium in lab

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Growth in - lab

  • Growth factors specific nutrients not required by all cells
  • Readily available in environment
  • No longer contain genes synthesize products
  • Ex: serum and amino acids
  • Axenic growth growth outside host cell
  • Pure culture single strain of single species
  • Liquid / broth medium
  • Organisms move about freely
  • Useful studying growth characteristics single species and growth kinetics / microbial biochemistry
  • Solid media
  • Useful for separating different organisms as found naturally in environment
  • Dilution streaking drop liquid culture cultured with inoculating loop and streaked across agar plate surface
  • Organism falls off loop as it moves along surface
  • Spread plate serial dilutions placed directly on surface separate agar plates
  • Isolate pure cultures
  • Stain from liquid culture
  • Early dilutions = confluent growth
  • Covers whole plate
  • Contains most bacteria
  • Later dilutions = individual colonies
  • Used enumerate viable bacteria original growth tube
  • Complex media possesses many preformed nutrients cells can readily assimilate
  • Nutrient rich but poorly defined
  • Include variety amino acids, peptides, nucleotides, vitamins and some sugar
  • Enriched media addition chemical building blocks cell otherwise have synthesize on own
  • Microbe grown hard characterize
  • Selective media favor growth one organism over other
  • Differential media expose biochemical differences between two species that grow equally well
  • Needed distinguish particular biochemical aspect

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Counting cells

  • Petroff Hausser Chamber
  • Directly counted under microscope
  • Dilution placed on slide (hemocytometer)
  • Forms precise volume
  • Organism counted within volume
  • Calculates concentration original growth
  • Living indistinguishable from dead
  • Fluorescence microscopy
  • Chemical dyes distinguish living / dead cells
  • Fluorescence activated cell sorter (FACS)
  • Electronic technique counts separate populations based on properties
  • Only counts cells synthesize fluorescent protein
  • Measures light scatter / intensity
  • Viable counts
  • Dilutions liquid culture subsequently poured into empty petri dishes and woled
  • Subsequent formed colonies counted
  • Doesn’t reflect cell size / growth stage
  • Usually underestimate number living cells
  • Biochemical assays
  • Measure dry weight culture
  • Cells collected by centrifugation
  • Optical density
  • Measure turbidity (cloudiness)
  • Uses light absorbance measures via spectrophotometer
  • Useful estimating population size
  • Must be corrected using standard curve

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Microbes Build Biomass

  • Heterotrophs rely on other organisms form organic compounds
  • Use as carbon source
  • Carbon disassembled generate energy
  • Reassembled make cell constituents
  • Converts large amounts organic carbon sources to CO2
  • Released into atmosphere
  • Autotrophs assimilate CO2 as carbon source and reduce it make complex cell constituents made up C, H, O
  • Later used by heterotrophs
  • Photoautotrophs = use light photosynthesis
  • Chemoautotrophs / lithotrophs = gain energy by oxidizing inorganic substances
  • Many microorganisms use both heterotrophy / autotrophy gain energy

Growth Cycle

  • Planktonic cells free living organisms that grow / multiply on own
  • Sent off by fixed location multicellular communities
  • Growth rate rate increase cell number / biomass
  • Proportional population size at given time
  • Generates exponential curve
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