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Classification of Microorganisms

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Ch 10<br />

<strong>Classification</strong><br />

<strong>of</strong><br />

<strong>Microorganisms</strong>


Student Learning Outcomes<br />

Define taxonomy, taxon, and phylogeny.<br />

List the characteristics <strong>of</strong> the Bacteria, Archaea, and Eukarya domains.<br />

Differentiate among eukaryotic, prokaryotic, and viral species.<br />

Explain the scientific naming<br />

Differentiate between culture, clone, and strain.<br />

Compare and contrast classification and identification.<br />

Explain the purpose <strong>of</strong> Bergey’s Manual.<br />

Describe how staining and biochemical tests are used to identify bacteria.<br />

Explain how serological tests and phage typing can be used to identify an<br />

unknown bacterium.<br />

Describe how a newly discovered microbe can be classified by ribotyping,<br />

DNA fingerprinting, and PCR.<br />

Describe how microorganisms can be identified by nucleic acid<br />

hybridization, DNA chips, and FISH.<br />

Explain and apply a dichotomous key<br />

Copyright © 2006 Pearson Education, Inc., publishing as Benjamin Cummings


Taxonomy and Phylogeny<br />

Taxonmoy: Science <strong>of</strong> classifying organisms. Provides<br />

universal names for organisms.<br />

• Taxonomic categories: Taxon / Taxa<br />

Phylogeny or Systematics: Evolutionary history <strong>of</strong> group <strong>of</strong><br />

organisms.<br />

• Taxonomic hierarchy shows phylogenetic (evolutionary),<br />

relationships among organisms.<br />

• 1969: Living organisms divided into five kingdoms.<br />

• 1978: Two types <strong>of</strong> prokaryotic cells found. Prokaryotic<br />

relationships determined by rRNA sequencing.<br />

• All Species Inventory (2001–2025)<br />

Copyright © 2006 Pearson Education, Inc., publishing as Benjamin Cummings


The Three-Domain System<br />

Foundation Fig 10.1<br />

Copyright © 2006 Pearson Education, Inc., publishing as Benjamin Cummings


Level Above Kindom: The Three-Domain System<br />

Carl<br />

Woese<br />

1978<br />

Eubacteria (virtuosos)<br />

Archaea (weirdoes)<br />

Eukarya (predators and thieves)<br />

Copyright © 2006 Pearson Education, Inc., publishing as Benjamin Cummings


Endosymbiotic Theory: Origin <strong>of</strong> Eukaryotes<br />

Figs 10.2, 10.3<br />

Copyright © 2006 Pearson Education, Inc., publishing as Benjamin Cummings<br />

Cyanophora paradoxa: modern example <strong>of</strong><br />

possible evolutionary process


Scientific Nomenclature<br />

Common names<br />

• Vary with languages<br />

• Vary with geography<br />

Binomial Nomenclature (genus + specific epithet)<br />

• Used worldwide<br />

• Escherichia coli<br />

• Homo sapiens<br />

Copyright © 2006 Pearson Education, Inc., publishing as Benjamin Cummings


Scientific Names<br />

Scientific Binomial<br />

Source <strong>of</strong> Genus<br />

Name<br />

Source <strong>of</strong><br />

Specific Epithet<br />

Klebsiella pneumoniae Honors Edwin Klebs The disease<br />

Pfiesteria piscicida Honors Lois Pfiester Disease in fish<br />

Salmonella<br />

typhimurium<br />

Streptococcus<br />

pyogenes<br />

Penicillium<br />

chrysogenum<br />

Trypanosoma cruzi<br />

Honors Daniel Salmon<br />

Chains <strong>of</strong> cells<br />

(strepto-)<br />

Tuftlike (penicill-)<br />

Corkscrew-like<br />

(trypano-, borer;<br />

soma-, body)<br />

Stupor (typh-) in<br />

mice (muri-)<br />

Forms pus (pyo-)<br />

Produces a yellow<br />

(chryso-) pigment<br />

Honors Oswaldo<br />

Cruz


<strong>Classification</strong>: Species Definition<br />

• Eukaryotic species: A group <strong>of</strong> closely related organisms<br />

that breed among themselves<br />

• Prokaryotic species: A population <strong>of</strong> cells with similar<br />

characteristics (Bergey’s Manual <strong>of</strong> Systematic<br />

Bacteriology is standard reference on bacterial<br />

classification).<br />

• Culture: Grown in laboratory media<br />

• Clone: Population <strong>of</strong> cells derived from a single cell<br />

• Strain: Genetically different cells within a clone<br />

• Viral species: Population <strong>of</strong> viruses with similar<br />

characteristics occupying a particular ecological niche.<br />

Viruses: not placed in kingdom nor domain – not<br />

composed <strong>of</strong> cells – cannot grow without a host cell.<br />

Copyright © 2006 Pearson Education, Inc., publishing as Benjamin Cummings


Phylogenetic Relationships <strong>of</strong> Prokaryotes<br />

Copyright © 2006 Pearson Education, Inc., publishing as Benjamin Cummings Fig. 10.6


Domain Eukarya<br />

• Animalia: Multicellular; no cell walls;<br />

chemoheterotrophic<br />

• Plantae: Multicellular; cellulose cell walls;<br />

usually photoautotrophic<br />

• Fungi: Chemoheterotrophic;<br />

unicellular or multicellular; cell walls <strong>of</strong> chitin;<br />

develop from spores or hyphal fragments<br />

• Protista: A catchall kingdom for eukaryotic<br />

organisms that do not fit other kingdoms<br />

• Grouped into clades based on rRNA<br />

Copyright © 2006 Pearson Education, Inc., publishing as Benjamin Cummings


Fig 10.8<br />

<strong>Classification</strong> and<br />

Identification<br />

<strong>Classification</strong>: Placing organisms in<br />

groups <strong>of</strong> related species. Lists <strong>of</strong><br />

characteristics <strong>of</strong> known organisms.<br />

Identification: Matching characteristics<br />

<strong>of</strong> an “unknown” to lists <strong>of</strong> known<br />

organisms.<br />

Copyright © 2006 Pearson Education, Inc., publishing as Benjamin Cummings


Identifying Bacteria<br />

Copyright © 2006 Pearson Education, Inc., publishing as Benjamin Cummings<br />

Applications,<br />

p. 283


Bergey’s Manual:<br />

Classifying and Identifying Prokaryotes<br />

Bergey’s Manual <strong>of</strong><br />

Determinative Bacteriology<br />

Provides identification<br />

schemes for identifying<br />

bacteria and archaea<br />

Morphology, differential<br />

staining, biochemical<br />

tests<br />

Bergey’s Manual <strong>of</strong><br />

Systematic Bacteriology<br />

Provides phylogenetic<br />

information on bacteria and<br />

archaea<br />

Copyright © 2006 Pearson Education, Inc., publishing as Benjamin Cummings<br />

Based on rRNA<br />

sequencing


Clinical Lab Identification<br />

Morphological<br />

characteristics<br />

Useful for identifying<br />

eukaryotes<br />

Differential staining<br />

Gram staining, acidfast<br />

staining<br />

Biochemical tests<br />

Determines presence<br />

<strong>of</strong> bacterial enzymes<br />

Numerical Rapid Identification<br />

Copyright © 2006 Pearson Education, Inc., publishing as Benjamin Cummings


Serology<br />

• Involves reactions <strong>of</strong><br />

microorganisms with<br />

specific antibodies:<br />

Combine known antiserum<br />

with unknown<br />

bacterium<br />

• Useful in determining the<br />

identity <strong>of</strong> strains and<br />

species, as well as<br />

relationships among<br />

organisms.<br />

Fig 10.10: Slide Agglutination<br />

• Examples:<br />

• Slide agglutination<br />

• ELISA (see lab)<br />

• Western blot (no details)<br />

Copyright © 2006 Pearson Education, Inc., publishing as Benjamin Cummings


Phage Typing<br />

Fig 10.13<br />

Identification <strong>of</strong><br />

bacterial species<br />

and strains by<br />

determining their<br />

susceptibility to<br />

various phages.<br />

More details on<br />

bacteriophages<br />

in Ch 13<br />

Copyright © 2006 Pearson Education, Inc., publishing as Benjamin Cummings


Genetics<br />

• DNA fingerprinting:<br />

Number and sizes <strong>of</strong> DNA<br />

fragments (fingerprints)<br />

produced by RE digests are<br />

used to determine genetic<br />

similarities.<br />

• Ribotyping: rRNA sequencing<br />

• Polymerase chain reaction<br />

(PCR) can be used to amplify<br />

a small amount <strong>of</strong> microbial<br />

DNA in a sample. The<br />

presence or identification <strong>of</strong><br />

an organism is indicated by<br />

Copyright amplified © 2006 Pearson Education, DNA. Inc., (see publishing as lab) Benjamin Cummings<br />

Fig 10.14: Electrophoresis <strong>of</strong><br />

RE digest <strong>of</strong> plasmid DNA


Nucleic Acid Hybridization<br />

Single strands <strong>of</strong> DNA or RNA, from related organisms<br />

will hydrogen-bond to form a double-stranded molecule;<br />

this bonding is called nucleic acid hybridization.<br />

Examples <strong>of</strong> Applications:<br />

• Southern blotting,<br />

• DNA chips, and<br />

• FISH<br />

Copyright © 2006 Pearson Education, Inc., publishing as Benjamin Cummings<br />

Fig 10.15


Nucleic Acid Hybridization: DNA Chip<br />

Fig 10.17


Fluorescent In Situ Hybridization (FISH)<br />

Add DNA or RNA<br />

probe attached to<br />

fluorescent dye<br />

for S. aureus<br />

Fig 10.18a–b


Dichotomous Key<br />

ANIMATION Dichotomous Keys: Overview<br />

ANIMATION Dichotomous Keys: Sample with<br />

Flowchart<br />

ANIMATION Dichotomous Keys: Practice

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