Lecture Exam 3 Flashcards
Bacterial growth
the process in which from a bacterial cell, two equivalent daughter cells are produced.
What are the four different phases in which bacterial growth can be modeled and what happens during each?
- Lag phase - bacteria adapt themselves to growth conditions (individual bacteria are maturing)
- Log phase or exponential phase - individual bacteria are reproducing at their maximum rate so their numbers increase.
- Stationary phase - the growth rate slows due to depletion of nutrients (bacteria begin to exhaust the resources that are available)
- Death phase - bacteria run out of nutrients and die.
On a standard growth curve the vertical axis is labeled as what? What does it stand for?
On the vertical axis growth is shown as L which = log (numbers) where numbers is the number of colony forming units per mL.
On a standard growth curve the horizontal axis is labeled as what? What does it stand for?
T (time)
What are three ways to directly measure bacteria?
- Spread plate technique (broth) cfu/mL
- Filtration
- Hemocytometer
What is a way to indirectly measure bacteria?
Turbidity
Sterilization
removal of all microbial life
Disinfection
removal of pathogens
Antisepsis
removal of pathogens from living tissue
Sanitation
lower microbial counts on eating utensils
What factors do the effectiveness of antimicrobial treatments rely on?
- number of microbes
- type of microbe
- environmental factors
- time of exposure
How do physical or chemical agents inhibit growth?
- effect membrane permeability
- damage proteins
- damage nucleic acids
What are the physical methods to inhibit bacterial growth (microbial control) and how? (be able to list 4-5)
- heat (autoclave, dry heat, pasteurization): moist heat denatures proteins; dry heat kills by oxidation
- low temperature: inhibits microbial growth
- filtration: removes microbes
- high pressure: denatures proteins
- dessication: prevents metabolism
- osmotic pressure: causes plasmolysis
- UV radiation: damages DNA
What are the chemical methods (types of disinfectants) used to inhibit bacterial growth?
- Phenols and phenolics
- Halogens (Iodine and Chlorine)
- Alcohols (ethanol and isopropanol)
- Heavy metals (silver, mercury, copper, zinc and selenium)
- Quaternary ammonium salts
- Aldehydes
- Gaseous Sterilizers
- Peroxygens
Sepsis refers to:
microbial contamination
Asepsis
the absence of significant contamination
What do aseptic surgery techniques prevent?
microbial contamination of wounds
What type of microbes are very difficult to destroy?
Endospores
What type of microbes vary widely in susceptibility to different methods of microbial control?
Vegetative pathogens
What type of environmental influences tend to inhibit antimicrobials?
the presence of organic material such as blood, feces, and saliva
What is Thermal Death Point (TDP)?
Lowest temperature at which all cells in a culture are killed in 10 minutes
What is Thermal Death Time (TDT)?
Time to kill all cells in a culture
What is an autoclave used for?
Steam sterilization
What does pasteurization do?
reduces spoilage organisms and pathogens
(E/C) What are some types of Phenols and Phenolics and what are their advantages?
- Phenol (carbolic acid) - used in some throat sprays and lozenges; acts as local anesthetic
- Phenolics - chemical derivatives of phenol
- Cresols - derived from coal tar (Lysol)
- Biphenols - effective against gram-positive staphylococci and streptococci; used in nurseries
ADVANTAGES: Stable, persist for long times after applied, and remain active in the presence of organic compounds.
(E/C) Are halogens effective alone or in compounds?
both
(E/C) What are the two types of Halogens?
Iodine (iodophores, betadine, isodine)
Chlorine (hypochlorous acid, sodium hypochlorite, chloramines)
(E/C) What is formed when chlorine is mixed with water?
hypochlorous acid
(E/C) What is chlorine easily inactivated by?
organic material
(E/C) What does alcohol do as a disinfectant?
- kills bacteria and fungi (not endospores or naked viruses)
- denatures proteins and disrupts cell membranes
(E/C) Why is alcohol not good for open wounds?
causes proteins to coagulate
(E/C) What are the two types of alcohol?
Ethanol - drinking alcohol
Isopopranol - rubbing alchol
(E/C) What are the categories of Heavy Metals used as disinfectants and what are they used for?
- Silver - protect infants against gonorrheal eye infections
- Mercury - disinfect skin wounds
- Copper - kill algae in pools and fish tanks
- Selenium - kills fungi and their spores; used for fungal infections and in dandruff shampoos
- Zinc - zinc chloride used in mouthwashes; zinc oxide used as antifungal agent in paints
(E/C) What are Quaternary Ammonium Compounds used for? What strains are resistant to them?
They are widely used surface active agents; they are cationic (positively charged) detergents; effective against gram positive bacteria (less effective against gram negative); destroy fungi, amoebas, and enveloped viruses.
Some Pseudomonas strains are resistant.
(E/C) What are the advantages and disadvantages of Quaternary Ammonium Compounds?
Advantages: Strong antimicrobial action, colorless, odorless, tasteless, stable, and nontoxic.
Disadvantages: Form foam. Organic matter interferes with effectiveness. Neutralized by soaps and anionic detergents.
(E/C) What disinfectant group includes some of the most effective antimicrobials? How do they work? What is an example of an aldehyde?
Aldehydes - inactivate proteins by forming covalent crosslinks with several functional groups.
Example: Formaldehyde
(E/C) What are gaseous sterilizers? What do they do? Give an example.
Chemicals that sterilize in a chamber similar to an autoclave - they denature proteins by replacing functional groups with alkyl groups.
Example: Ethylene Oxide (kills all microbes and endospores but requires exposure of 4 - 18 hours)
(E/C) What do peroxygens do? What do they do? Give an example.
Oxidizing agents that oxidize cellular components of treated microbes; disrupt membranes and proteins.
Example: Ozone
What are the top three most resistant types of microbes?
Prions (most), endospores of bacteria, mycobacteria
What are the top three least resistant?
Viruses with lipid envelopes (least), Gram-positive bacteria, Viruses without envelopes
Genomics
Molecular study of genomes
genome
All of the genetic material in a cell
gene
segment of DNA that encodes a functional product, usually a protien
genotype
genes of an organism
phenotype
expression of a gene
mutation
Change in genetic material - occurs when a DNA gene is damaged or changed in such a way as to alter the genetic message carried by that gene.
genetics
study of what genes are, how they carry information, how information is expressed, and how genes are replicated
Define the Central Dogma of molecular Biology.
the process by which the instructions in DNA are converted into a functional product. (“the flow of information in a cell”)
Explain the Central Dogma process.
The central dogma describes a two step process, transcription and translation by which the information in genes flows into proteins. (DNA - mRNA - Protein)
Transcription is the first step of gene expression, in which a particular segment of DNA is copied into RNA (mRNA) by the enzyme RNA polymerase.
Translation is the process in which cellular ribosomes create proteins. In translation, messenger RNA (mRNA) is decoded by a ribosome to produce a specific amino acid chain, or polypeptide.
How does genetic information flow between generations of cells?
The parent cells goes through replication and divides into two daughter cells.
How does genetic information flow between cells of the same generation?
through recombination to recombinant cells
Describe the structure and characteristics of DNA. (5 things)
- Polymer of nucleotides: adenine, thymine, cytosine, guanine (form sequence of 3 neucleotides (codons))
- Double helix associated with proteins
- “backbone” is deoxyribose-phosphate
- strands held together by hydrogen bonds between AT and CG
- strands are antiparallel
What does DNA stand for?
Deoxyribo Nucleic Acid
Nucleotide (NTS) = ?
PO4, deoxyribose, nitrogen base
Describe the structure and characteristics of RNA.
- single stranded
- transient
- mRNA serves as a template for ribosome’s to synthesize protein
What enzyme copies DNA (it is considered the main player)?
DNA polymerase
In which direction does DNA copy?
5 prime to 3 prime
Describe the steps of DNA replication.
- Enzymes unwind the parental double helix.
- Proteins stabilize the unwound parental DNA.
- the leading strand is synthesized continuously by DNA Polymerase in a 5 prime to 3 prime direction.
- The lagging strand is synthesized discontinuously in a 3 prime to 5 prime direction. RNA polymerase synthesizes a short RNA primer, which is then extended by DNA polymerase.
- DNA polymerase digests RNA primer and replaces it with DNA (okazaki fragment)
- DNA ligase joins the discontinuous fragments of the lagging strand.
What is a Okazaki fragment?
short, newly synthesized DNA fragments that are formed on the lagging template strand during DNA replication. They are complementary to the lagging template strand, together forming short double-stranded DNA sections.
What is the replication fork? What seperates the 2 strands of DNA?
the place where the parental double helix strand unwinds and becomes two strands - the leading strand and lagging strand.
- Helicase
- Topoisomerase
- single stranded DNA binding proteins (DNA Polymerase: binds to a sequence of DNA called the origin of replication)
What kind of replication is DNA replication? What does that mean?
semiconservative - produces two copies that each contain one of the original strands and one new strand.
What does DNA polymerase bind to in order to initiate synthesis?
double stranded nucleic acid
What are some basics of Transcription?
- DNA is transcribed to make RNA (mRNA, tRNA, and rRNA)
- begins when RNA polymerase binds to the promoter sequence. (promoter or TATA box, initiates transcription)
- proceeds in the 5 prime to 3 prime direction and does not need a primer
- stops when it reaches the terminator sequence
- uses nucleotides Adenine, uracil, cytosine, and guanine.
Explain exons and introns in transcription.
In most eukaryotic genes, coding regions (exons) are interrupted by noncoding regions (introns). During transcription, the entire gene is copied into a pre-mRNA, which includes exons and introns. During the process of RNA splicing, introns are removed and exons joined to form a contiguous coding sequence. This “mature” mRNA is ready for translation.
Explain the 5 prime cap and 3 prime poly A tail.
Eukaryotic pre-mRNA receives a 5’ cap and a 3’ poly (A) tail before introns are removed and the mRNA is considered ready for translation.
How does transcription and translation differ between eukaryotic and prokaryotic cells?
Transcription and Translation in Cells. In a prokaryotic cell, transcription and translation are coupled; that is, translation begins while the mRNA is still being synthesized. In a eukaryotic cell, transcription occurs in the nucleus, and translation occurs in the cytoplasm.
What are some basics of Translation?
- mRNA is translated in codons (3 nucleotides)
- Translation of mRNA begins at the start codon: AUG
- Translation ends at a STOP codon: UAA, UAG, UGA
What happens during the Initiation stage of translation?
(“beginning”): in this stage, the ribosome gets together with the mRNA and the first tRNA so translation can begin.
What happens during the Elongation stage of translation?
(“middle”): in this stage, amino acids are brought to the ribosome by tRNAs and linked together to form a chain.
What happens during the Termination stage of translation?
(“end”): in the last stage, the finished polypeptide is released to go and do its job in the cell.
Describe the detailed process of translation.
- components needed to begin translation come together (includes two ribosomal subunits, mRNA with codons, and tRNA with anticodons
- the ribosome assembles and on it, at the P site, a tRNA carrying the first amino acid is paired with the start codon on the mRNA.
- at the A site next to the P site on the ribosome a second codon of the mRNA pairs with a tRNA carrying the second amino acid
- The first amino acid joins to the second by a peptide bond, and the first tRNA is released.
- The ribosome moves along the mRNA until the second tRNA is in the P site, and the process continues and new amino acids are added to the polypeptide. (1000 per second)
- When the ribosome reaches a stop codon, the polypeptide is released.
- Finally, the last tRNA is released, and the ribosome comes apart. The released polypeptide forms a new protein.
Where does translation occur?
in the cytoplasm