Exam 3 Flashcards
Diseases are caused by transferable spore like (“seeds”) particles that could transmit infection (who?)
Fracastoro
Father of microbiology that 1st described microbes and invented microscope (who?)
Van Leeuwenhoek
Hand washing prevents childbirth fever (who?)
Semmelweis
Developed antiseptic in surgery (who?)
Lister
Disproved spontaneous generation. Also germ theory
Pasteur
Awarded Nobel prize for work on TB
Koch
Koch’s postulates (4)
Organism must be: 1 - found in all cases 2 - isolated and maint. in pure culture 3 - capable of reproducing infection 4 - retrieved from inoculated animal and recultured
What is unique about prokaryotes?
Do not have membrane bound organelles
Coccus is what shape
Circle/sphere
Bacillus are what shap
Rod
Spirillum are what shape
Spiral
Coccobacillus are what shape
Rod-shaped with round ends
Capsule or slime layer of bacteria is called
Glycocalyx
Gram positive exterior cell structure, color, charge
- Thick peptidoglycan layer surrounding cell membrane
- Stain purple
- Contain technical acids which create negative charge
Gram negative outer cell structure, and color and any other defining features
- Thin peptidoglycan layer surrounds cell membrane
- outer membrane (OM) surrounds peptidoglycan layer
- lipopolysaccharaide (LPS) embedded in OM composed of lipid A and O specific polysaccharide = endotoxins
- lipid A is toxic portion of LPS
- stain pink
Acid fast bacteria cell structure, color:
Cell wall with high lipid content that resists Gram staining
Note: requires unique dyes and heat to stain
Mycobacterium
Genus in which all species are acid-fast
E.g. M. Tuberculosis and M. Leprae
Cells that have NO cell wall exhibit a _______ cell structure because of a high ____________ content
Rigid; sterol
Sterol = waxy solids
Plasma membrane is the site of what activity
Oxidative phosphorylation for ATP synthesis
NOTE: only aerobic species (use O2)
Where are the enzymes that are used for DNA replication in a bacteria?
Cell membrane
What extrudes from cell membrane and acts as receptors in cell metabolism and cell communication?
Proteins
What is a mesome?
Invagination of cytoplasmic membrane that can form into vesicles and plays possible role in cell division
Also increases surface area
What is periplasmic space
Space between inner and outer membranes in Gram negative cells
What structures are the cytoplasm of bacteria?
Nucleiod
Plasmid (DNA)
Ribosomes
Storage granules
Region of bacteria cell containing DNA is called what
Nucleiod
Is there a nuclear membrane?
No
Structure inside the cell that is small extrachromosomal DNA with autonomous replication
Plasmid
Note: plasmids can be transferred between cells during conjugation and often carry antibiotic resistance genes
Organelles composed of ribosomal RNA and protein and are sites of protein synthesis are called what
Ribosomes
External structures of bacteria include
Flagella
Pili (aka fimbriae)
Capsule - slime layer, Glycocalyx
Endospore
What external structure of cell is used for locomotion
Flagella
Counterclockwise rotation of flagella produces ___ motion while clockwise rotation produces ___
Forward; tumbling
What serves as adherence factors on the external structure of a bacteria?
Pili/fimbrae
2 types of pili
1 - sex pili, transfer plasmids
2 - attachment pili, shorter, slow cell to attach to surfaces e.g. host tissue
What is a unique dormant cell type produced by some bacteria in response to adverse conditions?
Endospore
What is sporlation?
When conditions are not favorable cell forms endospore
What is germination
When conditions are favorable, returns to metabolically active cell (vegetative)
Why are endospores important (3)?
1 - allow cell to survive harsh environmental conditions
2 - high virulence factor (resistant to high temps e.g. boiling)
3 - play roll in botulism, tetanus, gangrene, anthrax
Father of taxonomy, classification system with 2 kingdoms (who?)
Linneaus
Classification system with 6 kingdoms (who?)
Woese
3 domain system due to comparison of DNA sequence. Name the 3
Bacteria
Archaea
Eukarya
Bacterial nomenclature:
King Puffs Chest. Oozing Footballs Gambles SavingS
Kingdom Phylum Class Order Family Genus Species Strain
What morphological characteristics (5) serve as identification markers for bacteria?
- Colony characteristics (what does the community look like?)
- Shape of cell
- Capsule? endospore?
- Staining (G+, G- or neither)
- Flagella - movement? (And arrangement of it)
Other than morphological characteristics, what are other ways (7) to tell bacteria apart from each other?
Biochemical testing - test for substrates e.g. sugars
Molecular taxonomy - size of genome, protein similarity, etc
Diagnostic molecular pathology
Polymerase chain reaction
Immunological tests -serotype indicates if it possesses specific set of antigens
Bacteriophage typing
Antibiotic sensitivity testing
An organism’s ______ indicates that it possesses a specific set of antigens
Serotype (immunological testing)
What is bacteriophage typing?
Phage is a virus that infects bacterium. Phages are host specific, so if you figure out what phage can infect a microbe, you can deduce the bacteria involved
Energy source of bacteria?
ATP (adenosine triphosphate)
Catabolism
Breakdown of carb, lipid, protein
Anabolism
Synthesis of cellular components
What metabolism of glucose occurs in the presence or absence of oxygen and 1 molecule of glucose is converted to 2 molecules of pyruvate?
Glycolysis
TCA cycle results in a net gain of:
2 ATP
6 NADH
2 FADH2
Glycolysis results in a net gain of
2 ATP
2 NADH
The final e- acceptor is what?
Oxygen
The use of reducing power of NADH and FADH2 to synth ATP is called
Oxidative phosphorylation
Chemiosmotic theory
ATP is synthesized as a result of proton motive force generated by passing electrons along e- transport chain
Aerobic respiration
Process of transferring e- from NADH and FADH2 to oxygen
Anaerobic respiration
Without O2, e- transferred to an inorganic terminal electron acceptor such as sulfur
Accumulations of highly molecular weight polymers that store glycogen and other energy resources are called
Storage granules
AKA inclusion bodies AKA granular inclusions
Fermentation
Absence of O2, e- transferred to organic terminal electron acceptor
Note: fermentation is used by organisms that can’t respire
What is the ATP yield from catabolism of glucose in aerobic respiration, anaerobic respiration, and fermentation?
Aerobic respiration = 38 ATP
Anaerobic respiration = 30 or less ATP
Fermentation = 2-4 ATP
What temperature do psychrophiles prefer?
-5 to 15˚ C
Psychro means “cold”
What temperature do mesophiles prefer?
25 - 45˚ C
E.g. E. Coli, in other words… they like human body temp which is 37˚C
Human body temp in C
37˚
What temperature do thermophiles prefer?
45 to 70˚C
E.g. hot springs and compost heaps
What temperature do hyperthermophiles prefer?
Over 70˚C
E.g. hot springs, boiling water
Oxygen requirement between obligate aerobes, obligate anaerobes, facultative anaerobes, microaerophiles and aerotolerant anaerobes
Obligate aerobes: need O2 Obligate anaerobes: killed if O2 Facultative anaerobes: better if O2 is around, but survives if not Microaerophiles: require small amt O2 Aerotolerant anaerobes: indifferent
Toxic derivatives of oxygen?
Superoxide
Hydrogen peroxide
OH
What requires O2 as final electron acceptor and uses O2 to generate energy?
Obligate aerobes
What uses fermentation or anaerobic respiration in absence of O2 AND will die in O2 environment?
Obligate Anaerobes
What grows best in O2 but can grow without it?
Facultative anaerobe
What requires small amounts of O2 to perform aerobic respiration?
Microaerophiles microbes
What can grow in the presence of O2 but does not use it for energy and uses fermentation (instead of aerobic or anaerobic respiration)?
Aerotolerant anaerobes
What lives and multiplies from pH 5-8
Neutrophils
What lives and multiplies at pH below 5.5?
Acidophiles
What lives and multiplies at or above pH 8.5?
Alkalophile
Osmotolerant bacteria
Tolerant of high salt environments (up to 10%)
What requires high levels of salt to live and multiply more than 10%?
Halophiles
A population of organisms descended from a single cell and separate from all other species
Pure cultures
Agar-agar is derived from what
Seaweed
Koch’s lab assistant created dishes that agar-agar is in, name
Petri dishes // Julius Richards Petri
After a bacterial cell gets big and duplicates itself, then it divides. This is called
Binary fission
It’s asexual cell division that happens frequently.
3 steps of binary fission
- Mesosome partitioned
- Septum
- 2 daughter genomes
Time it takes for a population to double in number is the
Generation time
Bacterial growth in laboratory conditions: in a closed system the population growth follows a pattern of stages called a
Growth curve
4 phases of growth curve
- Lag phase (latent) — “ramp up,” recruiting macromolecules & ATP
- Exponential phase (log) — cells divide all day, ‘er day
- Stationary phase — cells ate all their food and are chilling
- Death phase — population dies as they realize they really have no more food
How can you measure growth (2)
- Count them directly with microscope and hemacytometer
2. Count them indirectly with serial dilution
What is composed of 4 nucleotides? And what are the nucleotides?
DNA
Adenine, thymine, guanine, cytosine
What are the DNA nucleotide pairs?
A-T adenine-thymine
G-C guanine-cytosine
(Recall this is different from RNA which substitutes Uracil in place of Thymine)
A set of 3 nucleotides on a single strand (codon) encodes a specific
Amino acid
Replication of DNA begins at nucleotide sequence called
Ori
AKA Replication origin
Because replication goes in [____’ to _____’] direction on a DNA strand, it results in one strand being synthesized continuously called the ____________ strand. The strand that is synthesized discontinuously in pieces is called the ________ strand, and the pieces are referred to as _________ fragments.
5’ to 3’
Leading strand
Lagging strand
Okazaki fragments
How does gene expression work (central dogma)?
DNA —> RNA —> protein
Information passes from the genes to the RNA copy of the gene and the RNA copy directs the sequential assembly of a chain of amino acids.
RNA nucleotides are what?
A-U Adenine-Uracil
G-C Guanine-Cytosine
(Recall this is different from DNA which has Thymine instead of Uracil)
RNA moves along DNA to create ____ which is complementary to
mRNA; DNA
New mRNA is moved to ribosomes for translation.
Translation stops when RNA polymerase reaches
Stop signal on DNA
What is the process of synthesizing proteins that occurs more or less simultaneously with transcription? And where does this process happen?
Process: Translation
Location: Ribosomes
At the beginning of a gene called a _______, mRNA start doing its thing.
Promoter
Want to be able to switch on/off transcription. This is controlled by what? (This is unique to bacteria)
Operon
This is a group of genes that includes an operator, common promoter, 1+ structural genes that are controlled as a unit to produce mRNA
In humans, we have 1 promoter for each gene. Bacteria have 1 promoter for many genes.
What is an operon (group of genes - 3) in bacteria?
- operator
- common promoter
- 1+ structural genes that are controlled as a unit to produce mRNA
Bacteria frequently have 1+ gene encoded on 1 RNA using only 1 promoter, why?
Because transcription and translation are tightly linked
Operators/operons come in 2 flavors:
- Inducible operons: always OFF, must be turned ON
- Repressible: always ON must be turned OFF
Amino acids are assembled into growing ________ _______ that, when folded, make a protein.
Polypeptide chain
Inducible operon requires ________ to prevent a _________ protein from binding to operator.
inducer; repressor
And repressible operon requires the operator be bound by a repressor.
How do end products affect enzymes early in the gene transcription pathway to prevent production of end product?
Feedback inhibition
Spontaneous mutations vs induced mutations
Spontaneous mutations:
Not very common. Happens naturally.
Induced mutations:
— common, they are linked to UV light or radiation or exposure to chemical
What is the spontaneous mutation that involves an incorrect base being incorporated into the DNA during replication?
Base substitutions
Can lead to:
- point mutation (just 1 base is changed)
- missense mutation (substitution of different a.a. in protein) or
- nonsense (creates stop codon instead of a.a.)
What is the spontaneous mutation that involves removal or addition of nucleotides?
Deletion and insertion
In spontaneous deletion/insertion mutation, the risk is that you shift the condones/sequence and end up with a different mRNA and a different amino acid. This is called
Frameshift mutant
What is the spontaneous mutation where a segment of DNA spontaneously jumps from 1 site to another in the same OR different DNA molecules?
Transposons
AKA jumping genes
What are the (2) induced mutations?
Chemical mutagens — alters binding of DNA molecule
Radiation — UV (causes thymine dimers) and X-Ray (causes single and double stranded breaks in DNA)
What are the 3 types of spontaneous mutation base substitutions? Describe what happens in each.
Point mutation: 1 base change
Missense mutation: change out amino acid
Nonsense mutation: creates stop codon
UV light causes what?
Thymine dimers
X-ray causes what
Single and double stranded breaks in DNA
How do bacteria repair mutations (2)? Describe
Bacteria employ mismatch (excision repair) as well as SOS repair
Excision involves an enzyme cutting the damage DNA strand out and repairing the gap and then a ligase seals the “scar tissue” nick.
SOS repair is used for severely damaged DNA. It involves skipping over the damaged DNA
How best to study mutants?
Induce mutation and study specific types
Nutritional mutants: cells that grow without added growth factors
Prototroph
Proto “first”
Trophos “feeder”
self sufficient
Nutritional mutants: cells that grow ONLY WITH added growth factors from the lab
Auxotrophs
Auxillium “help”
Trophos “feeder”
Conditional lethal mutants are mutants defective for the synthesis of
Essential macromolecule under specific conditions (e.g. temperature)
Griffith noticed that some unknown compound was transforming bacterial cells and thus, transformation. Explain the phenomenon and what transformation is, as demonstrated by Griffith.
The unknown substance that was transforming bacterial cells was DNA.
So DNA is “naked” in the cell. When cell dies the DNA is released and taken up by recipient cells which causes a change in that 2nd cell.
Recipient cells that take up DNA must be described as
Competent
Many populations are naturally competent during what phase of growth?
Log phase (phase 2, exponential phase)
How can competence be induced (2)
Hint: think (1) chemically and (2) mechanically
CaCl2 treatment
Electrical current: electroporation
Conjugation requires contact. DNA transferred in 2 ways
1 - plasmid DNA
2 - chromosomal DNA
What is the process whereby transfer of bacterial genes from cell to cell via a bacteriophage?
Transduction
Bacteriophages can interact with cells in 2 ways
Lytic
Lysogenic
Lytic cells
Virus attaches to host and injects DNA. Sometimes they get right into the cell. Good guy host gets taken over by virus and will make a ton of viruses. So many virus bodies inside the host that the host cell bursts and dies… “avenge me!”
Lysogenic cycle
Zombies.
Does the same thing As lytic cycle in the beginning. However, this time the genetic material stays hidden. The good guy host unknowingly makes new cells with their own PLUS the virus’s genetics. And then the virus turns the switch ON and all the Sleeper Hosts turn into zombies.
There is lytic and lysogenic, so there are 2 possibilities for transduction. What are they?
Generalized — during production of making new viruses accidents happen. Sometimes only bacterial DNA is packaged. Still infects cells, but its just more bacterial DNA — nothing else happens.
Specialized — phage packages both viral AND bacterial chromosomal DNA. It packages specific chromosomal DNA near where the viral DNA integrated during lysogenic phase.
In generalized transduction, the phage serves as a
DNA vector (carries DNA)
What is the use of biological techniques to solve practical problems and produce useful products?
Biotech
Infection that literally means “brought forth by a healer”
Iatrogenic
Used to refer illness caused by or resulting from medical treatment
Iatros means “physician”
What kind of infection is a result of treatment in a hospital but is secondary to patient’s original condition?
Nosocomial infection
Nosokomos “person who tends the sick”
Sources of iatrogenic/nosocomial infection (3)
Touching people, blood, etc (direct)
Fomites (indirect)
Airborne transmission (indirect)
Ways to prevent and control iatrogenic/nosocomial infection (3)?
Disinfection/treatment with antiseptics
Hand washing
Monitoring patient population
Absence of all life
Sterilization
Killing/removing pathogens
Disinfection
Disinfectant applied to tissue
Antiseptic
Substance that kills/removes only bacteria
Bactericide
Severe infection leading to systemic immune response
Sepsis
Using aseptic technique
Asepsis
Aseptic techniques are employed to avoid microbial contamination E.g. boiling
Inhibition of microbe growth
Microbiostasis
Range of activity against microbes
Spectrum
mass murder
Drugs tested to determine lowest concentration at that inhibits microbe-called MIC (minimum inhibitory concentration)
Activity
Use of 1+ antibiotic to increase spectrum or killing effect
Combination therapy
Increased killing effect by using multiple antibiotics
Synergism
1 antibiotic interferes with action of another
Antagonism
Methods of sterilization (absence of all life)
Moist heat — steam, boiling water
Autoclave — steam with pressure
Dry heat — flame, oven
There are 5 classes of antibiotics, what are they?
- Inhibitors of cell wall synthesis
- Inhibitors of cell membrane
- Inhibitors of protein synthesis
- Inhibitors of nucleic acid synthesis
- Antimetabolites
Inhibitors of cell wall synthesis work by
Preventing cross-linking of peptidogycan by binding to enzyme receptor sites
Inhibitors of cell wall synthesis (5)
Penicillin Cephalosporins Bacitracin Vancomycin Isomiazid
Inhibitors of cell wall membrane (1) and how does it work
Polymyxin B & E; replace Mg2+ and Ca2+ from membrane lipids and distrusts structure of bacterial cell membrane
Inhibitors of protein synthesis (4)
Streptomycin
Tetracycline
Chloramphenicol
Erythromycin
Inhibitors of nucleic acid synthesis (2)
Rifampin — used for TB
Quinolones
Antimetabolites (1)
Sulfonamides ( Sulfa drugs)
E.g. used to treat mycobacterium leprae (leprosy)
Clinical problems with antibiotic use (5)
- symptoms cease, infection continues
- affects normal flora
- microbes become resistant
- toxicity possible
- sensitization - develop rashes, fever, anaphylaxis
What part of a bacteria enables it to survive harsh environmental conditions and makes it have a high virulence factor?
Endospores
High virulence means resistant to high temps e.g. boiling
How much O2 do microaerophiles require? What what does higher concentrations do?
2-10%; inhibit
What kind of mutant is defective for the synthesis of an essential macromolecule under specific conditions (e.g. temperature)?
Conditional lethal mutants
What would you induce in a bacterial cell if you gave it either a CaCl2 treatment or electrical current (e.g. electrophoration)
Competence in a recipient cell to be able to take up DNA
What antibiotic initially fights Staphylococcus aureus through cell wall synthesis?
Vancomycin
Damages kidney, nerve deafness, skin rashes, thrombophlebitis
What antibiotic is active against mycobacteria only through inhibition of cell wall synthesis?
Isoniazid
Inhibits synthesis of mycolic acids in cell walls of mycobacteria
What “vixen” bacteria replaces Mg2+ and Ca2+ from membrane lipids disrupting the structure of bacterial cell membrane?
Polymyxin B and E
Topical use only because its so toxic
What antimetabolite is a precursor to antibiotics, antimicrobial agents derived from sulfonic acid?
Sulfonamides (sulfa drugs)
What antimetabolite inhibits folic acid synthesis?
Sulfa drugs