Microbiology Exam 1 Flashcard

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Ribosomes

 

Prokariote

Eukariote

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Prokariote: 70S (50S & 30S)

 

Eukariote: 80S (40S & 60S)

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Positive Stain

 

Negative stain

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Posative stain: Stain the cell

 

Negative stain: Stain the background

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Gram staining process
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  1. crystal violet positive stain
  2. Iodine forms a complex with violet
  3. Alcohol used to wash away
  4. safranin used to counter stain (positive)

 

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Acid fast staining process
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  • Detect cells belonging to the genera mycobacteria or Nocardia
  • Gram stains don't work on these genera
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Gram + bacteria

A -> G

(1A, 2B, 2C, 1E, 1G)

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  • Actinomyces
  • Bacillus
  • Bifiidobacterium
  • Clostridium
  • Corynebacterium (& other diphtheroids)
  • Enterococcus
  • Gardnerella vaginalis

 

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Gram + bacteria

L -> S

(2L, 1M, 2P, 2S)

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  • Lactobacillus
  • Listeria
  • Mobiluncus
  • Peptostreptococcus
  • Propionibacterium
  • Staphylococcus
  • Streptococcus
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What determins cell shape?
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  • Murein Sacculus
  • Cytoskeleton
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Bacterial Parts

(all bacteria have)

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  • Cell Membrane
  • Genome
  • Ribosomes
  • Intracellular macromolecules
  • Almost always a cell wall
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Bacterial cell parts

(varies between genus & species)

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  • Plasmids
  • Pili
  • Capsules
  • Flagella
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Bacterial chromosomal storage
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  • Single circular chromosome of DSDNA
  • Extra info may be stored in plasmids or prophages.
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Cell surface structures
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  • Made of monomeric unit repeated N number of times
  • Type II T independent antigen (High variability, low cross reactivity) IGM, can't immunize
  • Flagella, Pilli, or capsules
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Flagella
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  • Protein
  • H antigen
  • Chemotaxis
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Common Pili (fimbriae)
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  • Protein
  • Adherence to host cell
  • Antiphagocytic
  • Motility
  • Sex pili for conjugation
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Capsules/ Slime layers/ Glycocalyx, Exopolysachuride
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  • K antigen
  • Polysachurides.  Sugar + Alcohol + D AA
  • Cell Surface associated
  • Water soluble slime layer
  • Or both
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Biofilms
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  • Glob of bacteria/yeast in mucoid exopolysachuride adhering to a wet surface
  • Present in > 1/2 all Bacterial infections
  • Bacteria phenotypically change when in biofilm.
  • MDR
  • Constant release of bacteria into blood
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Bacterial Cell membrane
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  • No sterols
  • site of respiration/make ATP
  • Site of septum formation for cell replication
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Periplasm structure
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(peptidoglycan)

  • polymer, NAM + NAG
  • NAM forms shoots & ladders connection
  • Human lysozimes break aminosugar threads connecting NAM to NAG
  • Role in nutrient uptake, structure, chemotaxis, osmoreceptors, degradative enzymes,
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LPS structure
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  • Lipid A: Disachurides, Toxic
  • Core: sugars
  • O AG chain: Sugars, highly interspecies specific, highly intraspecies variable.
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LPS virulence
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  • Broad specificity (effect many organs)
  • Endogenous & Exogenous Pyrogenic
  • Activates alternative & classical compliment
  • Activate hageman factor XIII
  • Heat stable
  • Doesn't form toxoids
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Pathway of LPS toxicity
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  1. LPS binds to human LBP in serum
  2. LPS-LBP interacts with CD14 receptors on membrane 
  3. signal transduction causes cell to release mediators

Soluble CD14 binds to endothelial cells, causes leakiness (hypotension) ARDS

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Sepsis, Severe sepsis, septic shock
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  1. systemic inflammation
  2. activation of coagulation
  3. inhibition of fibrinolysis
  • 1 and 2 results from LPS activating factor 13. 
  • Fibrinolysis is inhibited by plasminogen activator inhibitor
  • Sepsis survivors have immunosuppresion
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Virus replication cycle
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  1. Attachment
  2. Penetration
  3. Uncoating
  4. Macromolecular synthesis
  5. Assembly
  6. Release
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Persistant infections
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Latent: Intermittent acute episodes, between episodes, no viral particles present

Chronic: Virus present, disease absent

Slow: Long incubation time

 

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Protozoa
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Rhizopod: amoeba move with pseudopoda

Flagellates: Flagella

Ciliates: Cilia

Sporozoans: complex life cycle with sexual and asexual stages

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Basteria and temperature
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Mesophile: growth 20-55 C

Thermophiles: growth at > 55 C

Psychrophiles/ Cryophiles: growth <20 C

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Fermentation
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  • Occurs in cytosol
  • 1 2 3 or 4 carbon compounds generated, act as electron and proton acceptor turning NADH2 -> NAD+
  • Acidic carbon compound excreted from cell
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DNA synthesis
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DNA helicase: separates dsDNA -> ssDNA

Primase: makes short ssRNA primers for DNA synthesis

DNA gyrase: relieves wind tension

Topoisomerase IV: separates chromosomes

 

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Synthesis of peptidoglycan
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  1. Synthesis of amino sugar in cytoplasm.  UDP covalently bound to glucose as tag for converting into NAM/NAG.
  2. Assemble & move through cell membrane.  Performed by bactoprenol. 
  3. Attach new part to growing end of peptidoglycan by transglycolase enzyme
  4. Final cross linking by transpeptidase
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Bacterial growth cycle
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Lag phase: ^ cell volume & mass, DNA synthesis (no change in cell number)

Exponential phase: Balanced growth in size & number

Stationary phase: No net increase in cell number

Death phase: logarithmic cell death

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Transduction

Generalized

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  • defective phage particles of both lytic and temperate (must be lytic phase)
  1. Donor genome sheered
  2. Sheered genome + viral info packaged into virus "pseudovirons/abortive phages"
  3. Gene transferred
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Transduction

Lysogenic conversion

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  • Only by temperate bacteriophages
  • Lysogenic gives Bacteria new phenotype
  • (eg) code for toxin production

 

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Flexible gene pool
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  • Vary in size between bacteria species
  • Encode for extra functions for growth in selective media
  • Can horizontally be transfered
  • Genomic islands, prophages, plasmids
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Genomic islands
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  • Non endemic genes
  • can have flanking repeats, fragments of other elements, inserted in vicinity of tRNA sequences, unstable
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Sterilization
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Kill all

(although bacterial products still present)

  • Ethylene oxide
  • Aldehydes
  • Beta propionate
  • Peroxygens
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Disinfectant
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Killing most microorganisms

on inanimate objects

not spores

  • Chlorine
  • Phenol
  • Boiling
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Sanitization
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Clean and no pathogens

Removal of many bacteria

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Pasteurization
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Using heat to kill bacteria in liquids

below temperature of sterilization

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Antiseptic
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Chemicals used to kill microorganisms on skin surface or mucous membrane

  • Silver nitrate
  • Iodine (providone/betadyne)
  • Hydrogen peroxide
  • Anionic detergents SLS
  • alcohol
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Kirby-Bauer disk diffusion assay
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  • Qualitative
  • agar test for antibiotic susceptibility
  • test for inhibit growth (static/cidal)
  • block of antibiotic, lawn spread, observe growth

 

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MIC test

"Minimum inhibitory concentration:

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  • Quantitative
  • In vitro test for growth inhibition
  • diluted antimicrobial vs patient bacterial isolate, observe turbidity
  • Lowest effective concentration

 

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Metabolic Analog
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Isoniazide

Sulfa drugs

 

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Inhibition of cell wall synthesis
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Penicillins

cephalosporins

Carbapenems

Bacitracin

Vancomycin

Cyclosporine

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Inhibition of protein synthesis
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Chloramphenical

Aminoglycosides

Tetracyclines

oxazolidinones

glycyclines

Macrolides

Mupiricin

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Nucleic acid
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Sulfonamides

trimethoprim

fluoroquinolones/quinolones

rifampin

metronidazole

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alteration of cell membrane function
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polymixin

bacitracin

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MBC

"Minimum bactericidal concentration"

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  • Quantitative
  • kill 99.9%
  • Plate and grow bacteria from MIC to see if inhibited or killed
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Antimicrobial Seruncidal concentration
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  • Quantitative
  • concentration needed to kill bacteria in presence of serum
  • Blood test to determine if MBC was accomplished
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