Exam 1 Cards Flashcards
What temperature does an incubator need to be for growth?
Any temperature that provides optimum condition for a specific microbe to grow
Describe glycocalyx function
Gelatinous outermost layer of cell (usually)
Functions:
- Protect cell from drying out (desiccation)
- Adherence to surfaces or other cells
- Strengthens cell
- Composition (polysaccharides or glycoproteins)
Bacteria and Archaeal note:
- Can hide them from immune system because humans have similar composition
What are the challenges of culturing viruses?
They require a live host to replicate, so you are always co-culturing viruses and their hosts
- In-vivo uses the whole animal host
- In-vitro in just the cells/tissues
Bacterial species vs. strain/subspecies
Species:
- Group of cells that share a similar pattern of traits
- 70-80% genomic similarity
Strains/subspecies:
- Also 70-80% genomic similarity but have different characteristics
- ex. the various Staphylococcus species and subspecies (due to various antibiotic reactions/growth/appearance)
Explain the 3 animal cell “entry” methods
- ) Direct penetration: (only naked viruses) capsid binds to host cell membrane, sinks in to create a pore, which the genome enters through. Empty structure is left behind on surface
- ) Membrane fusion: viral envelope and cell membrane attach and fuse, releases capsid into cell cytoplasm, uncoats and releases genome. Viral envelope’s glycoproteins remain as part of host membrane
- ) Endocytosis: after attachment, viral glycoproteins trigger host cell to engulf the entire virus. Membranes/envelope then removed and uncoating of capsid releases genome
Label microscope type using image.
Briefly describe the formation of an endospore
- DNA is replicated
- Cytoplastic membrane surrounds daughter DNA creating a forespore
- A second membrane surrounds the forespore to create double membrane
- Peptidoglycan layers fill the space between the double membrane (called “Cortex”)
- Spore coat (proteins) is created around double membrane (resistant to heat/chemicals)
- Endospore is released and mother cell dies
Algae characteristics?
- Eukaryotes
- Can be unicellular or multicellular
- Photosynthetic (autotroph)
- Cell walls made of: agar or calcium carbonate
Taxonomy is?
Science of putting organisms into categories/classifying them
Explain the effects of water on microbes
Pressures exerted on cell
- osmotic (related to concentration of solutes)
- hydrostatic (related to depth)
Water also dissolves enzymes and nutrients
What concepts are important in microscopy?
Microscopy is the magnification of an object using light or electrons
Concepts:
- Wavelength
- Magnification
- Resolution
- Contrast
What are the types of “probe microscopes”?
- Scanning tunneling
- Atomic force
What is Bioremediation?
Using living microbes to detoxify polluted environments
How do you isolate a colony of microbes?
With solid media, Petri dish, and an inoculating loop. Use the T-streak method to dilute single bacterium so they can form colonies across different areas on a Petri dish.
How do you inspect and identify when culturing microbes?
Inspect colony morphology (shapes) and identify by:
- Appearance
- Metabolism
- Growth requirements (ex. carbon sources, pH, oxygen needs, etc.)
- Energy requirements
- Genetic makeup and immunological characteristics
Differential stain examples?
- Gram stain
- Acid-fast stain
- Endospore stain
Describe -ssRNA synthesis
Genome replication:
- ”–ssRNA” viruses carry “RNA-dependent RNA transcriptase” in capsid, released during uncoating
- This enzyme transcribes the -ssRNA into +ssRNA
- Newly created +ssRNA used as template to create more -ssRNA (and to translate proteins)
Protein synthesis:
- Host ribosomes translate proteins directly from the +ssRNA
Define: anaerobic, aerobic, facultative anaerobe
Anaerobic - grows without oxygen
Aerobic - grows with oxygen
Facultative - can grow with or without oxygen
Fungi characteristics?
- Non-autotrophic
- Eukaryotic
- Cell walls composed of chitin
(Mold and Yeast)
What are some structures that helminths use for attachment?
Suckers, hooks, teeth
Fermentation and pasteurization concepts
Fermentation - process that converts sugar to an alcohol or acid
- (Ex. Yeast converting sugar to alcohol, or bacteria converting sugar to acid)
Pasteur invented pasteurization to remove acid producing bacteria from wine
Buchner showed chemical reactions required enzymes, not living cells
What type of microscope will we be using in lab?
Compound light microscope
What helps facilitate biofilm attachment to surfaces?
- Extracellular Matrix (ECM)
- Pili and fimbreae
- Other proteins
What allows “archaeal” attachment or adhering?
- Fimbriae: much shorter than flagella, attachment and twitching
- Hami: hook structures on some fimbriae used for attaching to surfaces (only Archaea)
Review Table 4.2 in textbook. Study guide on reverse side of flashcard.
Important to know (from lecture video):
- Source of magnification (light, electrons, etc.)
- Type of specimens they can see (bacteria, virus, DNA, etc.)
- Specimens living or dead?
- Stain usage needed?
How can you improve a poor Bright-field microscope image?
A light background with light images and poor contrast can be improved with dyes
What is osmosis?
- Movement of water molecules directly across membrane
- Type of passive transport
What is transduction?
Host DNA is packaged into a viral capsid and transferred into a new host cell
What are the 3 major types of microscopes?
- Light
- Electron
- Probe
What are mycoplasma?
Bacteria without cell walls that are sensitive to osmotic pressure (can burst)
What are the shapes of virions?
Helical: tube-like
Polyhedral: soccer-ball like sphere
Complex: all other shapes (T4 bacteriophage is made of both, helical and polyhedral shapes, therefore would be considered complex shape)
How are DNA viruses synthesized in animals?
DNA viruses use process very similar to normal dsDNA replication:
- Host cell DNA polymerase replicates the viral dsDNA
- Host cell RNA polymerase transcribes DNA to create mRNA for translation of proteins
(Since normal DNA is double-stranded, ssDNA folds back on itself pretending to be dsDNA for replication and then released as ssDNA)
Describe a biofilm
Community of different microbe species living on a surface (typically antibiotic resistant and/or immune resistant).
Biofilm can also hide microbes from immune system.
What are Peripheral Proteins?
Proteins that are attached to one side/one half of phospholipid bilayer
Classify bacteria by pH levels
Neutrophiles - grow in neutral conditions
Acidophiles - lower pH
Alkalinophiles - higher pH
- Bacteria 6.5-7.5 pH
- Yeasts 5-6 pH
What is a physical difference between bacteria and archaea?
Archaea does not contain peptidoglycan in its cell walls
Describe acid-fast stains (a differential stain)
Identifies mycobacteria with waxy cell walls (mycolic acid, hard to stain).
- Carbolfushin (pink dye #1) stains all bacteria
- Acid alcohol (decolorizes non-waxy bacteria)
- Methylene blue (#2 dye) stains non-waxy bacteria
Pink = waxy Blue = non-waxy
(Bonus: mycobacteria cause many human diseases)
Manipulation of the genetics of microbes, plants, and animals, for the purpose of creating GMOs
Genetic engineering
What are hopanoids?
- Cholesterol-like molecules, enable bacterial cytoplasmic membrane to be fluid and be in extreme high or low temperatures.
- Important in ensuring transport during these extreme temperatures.
Fungi morphology?
- Some yeasts look similar to bacteria
- Some yeasts grow in loosely attached filaments
- Some fungi form “hyphae” (long threads) woven together to form “mycelium”
- Fungi can be “dimorphic” (different morphology based on growth conditions)
Describe endospore stains (a differential stain)
Used to distinguish between endospore and vegetative bacteria.
- Boil in Malachite (green dye #1) stains endospores
- Water (decolorizes vegetative)
- Safranin (pink dye #2) stains vegetative
Green = endospores Pink = vegetative
(Bonus: endospores can survive extremely harsh conditions and resist stains)
What are prions?
Infectious protein particles that lack DNA/RNA that affect animals.
Ex.
- PrP is a cytoplasmic membrane protein anchored in rafts
- Normal cellular PrP structure: helices
- Prion PrP contains pleated-sheets that cause cellular PrP to misfold and turn into prion-PrP
- Misfolding proteins causes holes in tissue
Describe capsid (protein coat) functions
- Provides protection for nucleic acid
- Attachment to host cell (if no envelope)
- Made of: capsomeres
- Determines shape
What magnification do you need to see bacteria?
1,000x (using oil immersion lens which allows more light to enter objective lens)
Bright-field microscope details?
- Light microscope
- Simple (1 lens) or compound (more than 1 lens)
- Light background, dark specimen
- Total magnification = objective lens x ocular lens magnification levels (i.e. 4 x 10 = 40x)
- Most have condenser lens to focus light through specimen
Describe bacterial endospores characteristics
- Unique structures produced by vegetative (metabolically active) bacteria.
- Bacterial endospores are not metabolically active to be protected against unfavorable conditions (lack of nutrients or extreme physical).
- Once conditions become favorable, the spores will germinate and become active again
- Spores can release deadly toxins that cause fatal diseases like anthrax, tetanus, and gangrene
(Bonus: DNA is protected/preserved by dipicolinic acid and calcium ions)
What are some cytopathic effects of viruses?
Viruses can alter or damage host cells:
- Change shape
- Damage intracellular components
- Lysis
Accumulated damage kills most hosts
Name some of Koch’s contributions
Found rod-shaped bacterium in blood of Anthrax diseased animals. The spore form caused disease.
Developed microbiology techniques:
- simple staining
- first photomicrograph of bacteria and bacteria in diseased tissue
- technique for estimating CFU/ml (colony forming unit)
- use of steam to sterilize media
- use of Petri dishes
- techniques to transfer bacteria
- bacteria as a district species
Examples of endospore forming bacteria that cause diseases?
Bacillus anthracis - agent of anthrax
Clostridium tetani - cause of tetanus
Clostridium perfringens - cause of gangrene
Clostridium botulinum - cause of botulism
Clostridiodes difficile - “C. diff” gastrointestinal disease
Describe special stains
Negative stains:
- Reveals capsules (surrounding bacteria) by staining background with acidic dye
- Then counter-stains used on bacteria
- Capsules remain colorless and will appear as halo
Flagellar stains:
- Increases contract and thickness of flagella appendages on bacteria
- Allows light microscope to see them
(Bonus: capsules can indicate pathogens and remain colorless due to negative charge)
What are the types of “light microscopes”?
- Bright-field
- Dark-field
- Phase
- Fluorescence
- Confocal
A method for obtaining a pure culture (culture composed of a single bacteria) from a patient?
T-streaking
Describe the structure of virions
Nucleic acid: DNA or RNA, double or single stranded
Capsid: protein coat that surrounds the nucleic acid
Envelope (some viruses): phospholipid membrane (for protection and recognition site for host cells)
(Viral genomes are very small and usually only contain just enough information for replication and nothing else.)
Group bacteria based on the different types of water pressure.
Obligate halophiles - require higher osmotic pressure, up to 30% salt
Facultative halophiles - indifferent to osmotic pressure (2-15% salt)
Barophiles - live under extreme hydrostatic pressure
What microbes do not require an electron microscope to see?
Bacteria, archaea, fungi, protozoa, and algae
What are Integral Proteins?
Proteins that run all the way through cytoplasmic membrane
Hypertonic solution
- More solute (higher concentration) on outside
- Water will flow outward of cell
Mold and yeast fall into this microbe type
Fungi
Are all viruses considered human pathogens?
No, bacteriophages are examples that do not cause harm to humans
Define: culture, medium, inoculum, inoculation
Culture: growth of microorganisms (both the verb and noun)
Media: nutrients for the growth
Inoculum: small sample of microbes
Inoculation: process of putting inoculum onto media to grow
Explain the basic steps in animal virus replication
1.) Attachment: like bacteriophages, viral proteins and host cell receptors facilitate attachment. However, animal viruses use glycoprotein spikes or other attachment molecules. (no tails/tail fibers)
2a. ) Entry: direct penetration, membrane fusion, or endocytosis
2b. ) Uncoating: viruses that enter with capsid or envelope need to remove it to release genome
- ) Synthesis: different strategy and location based on genome type (DNA vs. RNA; ss vs. ds)
- ) Assembly: DNA viruses in nucleus, RNA viruses in cytoplasm (usually)
5a. ) Release: enveloped viruses released by budding (taking along portions of cellular membrane coated with viral glycoproteins…which then turns into the envelope)
5b. ) Release: naked viruses released by exocytosis or lysing and killing the cell
Are cell walls present in Eukaryotes?
Yes, only in fungi, algae, and plants
Fungi cell walls = chitin or glucomannan
Algae cell walls = agar, algin, or calcium carbonate
Plant cell walls = cellulose
(No cell walls in animals, protozoa, or helminths)
Why is the synthesis strategy for DNA viruses different than RNA viruses?
- Eukaryotic genomes are made of DNA, and not RNA
- DNA viruses usually replicate in nucleus
- RNA viruses usually replicate in cytoplasm
Lipopolysaccharide (LPS, outer membrane) importance?
LPS only in gram negative bacteria
Understanding LPS structure:
- O-antigen (outermost part of cell): body’s response to antigen helps identify
- Core sugars: help identify genus or species
- Lipid A: endotoxin released by dying bacteria can cause shock
Physical requirements for growth?
- Temperature
- pH
- Water
Isotonic solution
- Same concentration of solute inside and outside
- No net flow of water
Steps to prepare sample for staining?
Coloring specimen with dye:
- add thin film of microbe to slide (AKA a “smear”)
- “fix” (attach) to the slide with heat, air, or chemicals
Some reasons to culture viruses?
- Isolate
- Identify
- Prepare vaccines
- Perform research on replication, genetics, effects on hosts
What is a Uniport?
Active transport protein that helps one molecule to cross membrane in one direction
What are Cilia?
Short hair-like protrusions used for movement (typically on Protozoa)
How can we classify bacteria based on optimum growth temperature?
Psychrophiles - low temp, below 20C
Mesophiles - medium temp, 20-40C (most pathogens)
Thermophiles - high temp, above 45C
Hyperthermophiles - extremely high, above 80C
What allows “bacterial” attachment or adhering?
- Fimbriae: much shorter than flagella, attachment and twitching
- Pili (only 1-2 per cell): only found in Bacteria…special type of fimbriae for DNA exchange or twitching
Hypotonic solution
- Less solute (lower concentration) on outside
- Water flows inward to cell
What does it mean to be autotrophic?
Organism that produces its own food using light, water, carbon dioxide, or other chemicals.
(Photosynthesis is a type of this)
How do cavities in teeth form?
Biofilms form on teeth, they absorb sugar as nutrients, and then secrete acid that eats away at tooth enamel
This microbe grows in extreme conditions
Archaea
When a virus is extracellular (outside a host), what is it also known as?
A virion
Helminths characteristics?
- Include tapeworms, flukes, roundworms
- Microscopic when larval stage
- Parasitic helminths spend part of their lives in GI tract
- Reproduce sexually in host’s body
Brief description of a taxonomic system?
Giving living things a scientific name, and classifying/grouping by structure and order.
Binomial naming (always in italics): - “Genus” (first letter Capitalized) then “species” (all lowercase)
Name some characteristics of viruses
- Acellular/non-living infectious agents containing genetic material (DNA or RNA)
- Replication method depends on type of host (bacteria, animal, etc.)
- Lack cytoplasmic membrane
- Do not grow, respond to environment, or independently carry out metabolism
- Cannot reproduce independently
Describe some other “light microscopes” (besides Bright-field)
Dark-Field:
- Observes pale objects
- Dark background, light specimen
- No condenser lens (scatters light to create darker background)
Phase:
- Examines living organisms
- In / out of phase light produces contrast in image
Florescent:
- Fluorescent dyes “glow in the dark” under UV light
- Can be used for labelling specific things
Confocal:
- Useful for biofilm study
- Also fluorescent dyes, but use UV lasers instead