Classification of Bacteria & Diagnostic Methods Flashcards
Terminology
- ______: Process by which populations of organisms become altered over relatively long periods of time. Organisms may divide into separate groups or ______, combine to form ______ organisms, or cease to exist due to _______
- ______: The study of evolutionary relationships among organisms by comparing gene sequencing data and morphological data matrices
- _______: The assignment of organisms into groups within a system of shared qualities or characteristics
- ______: The classification of organisms based on phylogenetic relationships of groups of organisms, rather than on shared physical features as used in ______Classification
- ______: The study of classification of organisms in order to reconstruct their evolutionary history and relationships
- _____ ______: The description, classification, identification, and naming of organisms where they’re grouped into hierarchical units known as a ______
- _______: The act or process of identifying or determining the nature and cause of an infection or infectious disease
- Evolutions; branches; hybrid; extinction
- Phylogenetics
- Classification
- Cladistics; Linnean
- Systematics
- Linnean Taxonomy; taxon
- Diagnostics
Nomenclature: Carolus _______(1707-1778)
- Swedish botanist and physician
- Developed the ______ System of _______ (Taxonomy) based on shared _______ characteristics (genus & _______)
- Linnaeus
- Binomial; Nomenclature; physical; species
Taxonomic Ranking
- ________: 3 exist
- _______: 6 exist
- _____/______
- ______
- ______
- _______
- _______
- ________
- Domain
- Kingdom
- Phylum/Division
- Class
- Order
- Family
- Genus
- Specis
6 Kindoms:
- ______(_____): All prokaryotic cells including _____, achaea_______, & cyano______
- ______(____): Unicellular non-photosynthetic including ___-_____ algae, _____, and _____
- _______(_____): Photosynthetic algae with chlorophylls a and c
- ______
- _____: Non-photosynthetic including _____, _____, and ______
- ________
- Bacteria (Monera); bacteria; archaeabacteria; cyanobacteria
- Protozoa (Protista); non-photosynthetic; ameoba; ciliates
- Chromista (Protista)
- Plantae
- Fungi; molds; yeast; mushrooms
- Animalia
Taxonomic Ranking of Bacteria
- Have a sub-category of Species called ______ typically indicated by a combination of ______ and _____ about where it was ______
- First step in identification is to examine a _____ _____ under a microscope
- strain; letters; numbers; isolated
- Pure culture
Light Microscopes
- Magnification up to _____x
- _____ _____: Uses light to illuminate an object that absorbs light in denser areas to give contrast for visualization, and requires _____ due to the little difference between the _______ index of bacteria the surrounding liquid
- _____ _____: Excludes the non-scattered light from the field of view leaving the background dark and the object in the field of view light, making it good to view very ______ or _____ species
- _____ _____: Phase shifts in light are converted to brightness in the field of view allowing normally _______ structures to be seen, good for identifying different _____ in bacteria
- ______ ______ ______(___): Uses interferometry rather than phase shifts to allow visualization of invisible structures in cells
- ______: Uses a mercury lamp and filter to illuminate the viewing field with light of a specific wavelength, a second filter removes that wavelength but allows the light emitted from an excited molecule (dye) to be visualized
- 400
- Bright field; staining; refractive
- Dark field; thin; pale
- Phase contrast; translucent; densities
- Differential Interference Contrast (DIC)
- Fluorescence
Confocal Microscopy
- Magnification up to _______x
- Uses a spatial pinhole place at the confocal plane of the lens to eliminate out-of-focus light enabling the reconstruction of ___-______ from the obtained images
- Done more often with ______ cells than bacteria
- 17,000
- 3D-structures
- Eukaryotic
Electron Microscopy
- Magnification up to ________x
- Most _______
- Uses a beam of accelerated _______ as a source of illumination
- _______(___): Negative stains will stain the cell on a copper grid and can be viewed directly by taking a thin ______ through the cell allowing for _____ components of the cell to be viewed
- _________(____): Probing the specimen with a focused electron beam that is scanned across a rectangular area of the surface of the specimen
- 500,000
- Powerful
- electrons
- Transmission (TEM); slice; internal
- Scanning (SEM)
Gram Stain: Hans Christian Gram (1853-1938)
-Gram Positive bacteria appear ____/____ in color
-Gram Negative bacteria appear ____/____ in color
Steps:
1. Create a ______ on a glass slide and let it _____ dry
2. _____ bacteria to the slide
3. Bacteria are stained with _____ ____, making all bacteria appear blue/purple
4. Bacteria get treated with a ______ solution, this fixes the ____ ____ into the cell walls of Gram ______ bacteria
5. Cells are washed with an ______-_____ mixture, this washes the _____ _____ out of Gram _______ bacteria
6. Counterstain the slide with _____, this turns Gram ______ pink/red
- purple/blue
- pink/red
1. smear; air
2. Fix
3. crystal violet
4. iodine; crystal violet; Positive
5. acetone-alcohol; crystal violet; Negative
6. Safranin; Negative
Acid Fast (Ziehl-Neelsen) Stain
Steps:
1. Create a ______ on a glass slide and let it ____ dry
2. ______ bacteria to the slide
3. Bacteria are stained with ___________
4. Apply _____ to steam the bacteria, this stains all bacteria ______
5. Wash the bacteria with an ___-_____ mixture, this washes the stain out of all ___-_____ ____ cells
6. Counter stain with ______ _____, this stains all non-Acid Fast bacteria ______
- smear; air
- Fix
- carbolfuschin
- heat; red
- acid-alcohol; non-Acid Fast
- methylene blue; blue
Acid-Fast Bacteria
- Most are members of the genus _________
- The most important species for dentists is __________ __________
- Often use _______ from a patient’s cough to check for the presence of _______ ______
- Mycobacterium
- Mycobacterium tuberculosis
- sputum; Mycobacterium tuberculosis
Bacterial Shape
- ______
- Curved _____
- _________
- ______: Bacteria is wider in the center with pointed ends
- ______: Scanning and thin-section EM discovered in Egypt
- Rods
- rods
- Fusiform
- Square
-_______ are clonal representations of a bacteria
-Colonies
- _________ are mobile bacteria so the colony spreads making it appear layered
- _______ are mucous like
- __________ _________ (__) will form a characteristic star-shaped colony
- ________ _______ will take on a different appearance depending on the environment and conditions in which it is grown
- Proteus
- Klebsiella
- Aggregatibacter actinomycetemcomitans
- Staphylococcus mutans
Bacterial Pigments
- Serratia marcesenes produces a unique pigment called _________, giving it a bright ____/_____ color
- Black-pigmented _______ species produce melanin, and are oral bacteria useful for diagnosis
- prodigiosin; red/orange
- Bacteroides
Generalizations of Oral Bacteria
- Almost all Gram-positive cocci are _______ species
- Almost all Gram-Negative spirochetes are ______ species
- Almost all Gram-Negative cocci are ________ species
- ALL Gram-Negative bacteria with star colony morphology are __________ ________
- ALL All Gram-Negative rods with brown-black pigmented colonies are ________ species and ______ _______
- Streptococcus
- Treponema
- Veillonella
- Aggregatibacter actinomycetemcomitans
- Bacteroides; Porphyromonas gingivalis
Oral Bacterial Enzymes Useful for Diagnostics
- _______ test to see if the species contains _______ that can degrade hydrogen peroxide to water and oxygen via the production of oxygen gas bubbles
- ____________ ____________ is ______ positive
- Catalase; catalase
- Aggregatibacter actinomycetemcomitans; catalase
Other Bacterial Products Used for Diagnostics
- End-products of _______, like the production of gas
- ______ profiles from sugar fermentation
- _____ _____ profiles, via gas-liquid ________
- _______ profiles by isolating them via SDS-Page, not enzymes
- ________ profiles
- fermentation
- Acid
- Fatty acid; chromatography
- Protein
- Antigen
Differential and Selective Growth
- ________ ______: Contains specific ingredients or chemicals that help to differentiate species that can or can’t carry out a certain function.
- _________ Agar tests for the ability to ferment lactose, _____ ______ is lactose positive, and ______ _____ is lactose negative.
- ______ Agar tests the ability of bacteria to lyse RBCs. _-______: Complete _______ of RBCs, characteristic of Streptococcus pyogenes. __-_______: Partial _______ of RBCs resulting in a greening of the agar, characteristic of Escherichia coli. __-_____: No ______ of RBCs at all, characteristic of Staphylococcus epidermidis
- _____ _____: Selects for one microorganism to grow while actively selecting against the growth of another
- Differential Media
- MacConkey’s; Escherichia coli; Enterobacter aerogenes
- Blood; β-hemolysis; hemolysis; α-hemolysis; hemolysis; γ-hemolysis; hemolysis
- Selective Media
-________: Based on metabolic characteristic like fermentation reactions
-________: Based on immunospecific reactions, Ag- Ab interactions
Methods that use Ag-Ab Reactions to Identify Bacteria:
-________ Blotting
-_______-_______ ______ _______(____)
-__________
- Biotype
- Serotype
- Western
- Enzyme-linked Immunosorbent Assay (ELISA)
- Immunofluorescence
- Most capsules are composed of ______, but can be made of ______
- ______ and _____ _____ are poor antigens
- Carbohydrates; proteins
- Lipids; nucleic acid
- _______ _______: The method of separating the components of serum
- ________: Are the gamma globulin component of the serum that can recognize specific ______ on target antigens
- _______: Can be a whole bacteria/virus or their protein products
- Paper Electrophoresis
- Antibodies; epitopes
- Antigens
Application of Ag-Ab Reaction
Enzyme-linked immunosorbent assay (ELISA):
1. Coat _____ on microplate
2. Allow _____ to bind to the plate and block unoccupied sites with _______ protein
3. Add _____ to each well
4. Add _______ _________ Ab to each well and develop colorimetic reaction with the appropriate substrate
5. Read _________ in the spectrophotometer with the appropriate filter and quantify relative Ag levels
- Antigen
- Antigen; neutral
- Antibodies
- Conjugated secondary
- absorbance
Methods of Diagnosing Infectious Disease
- Detect the presence of a specific ________
- Detect the presence of _____ specific for a particular _______, also indicates you’re infected with the disease
- pathogen
2. antibodies; pathogen
-________: Analytical approach based on DNA sequence analysis
Methods:
-_____ ____ gene sequencing to identify species
-______ _____ _____(___) to identify a gene
-_______ ______ allows for you to compare a sequence to a sequence bank to identify the organism
-________: DNA/RNA is taken from an area, sequenced, and then the total number and type of organisms present can be determined
- Genotyping
- 16S rRNA
- Polymerase Chain Reaction (PCR)
- Genomic Sequencing
- Metagenomics
Key Developments in Bacterial Classification and Diagnostics
- ______ ______ ______(__) ___: Inoculate the bacterial sample into each bubble and observe the results, you then compare the pattern found to known species
- Bergey’s Manual of _______ _______: Provides the species by name, what sugars it can/can’t ferment, enzymes produced, GC content, and DNA melting point to help for identification (mostly ______ information). AKA the Bible of _________
- Automated Phenotypic Identification (API) 20E
- Systematic Bacteriology; phenotypic; Identification
- _________- indicates chains
- _________- indicates clusters
- Strepto
- Staphylo
- Eikenelia corrodens is a _______ rod
- Neisseria gonorrhoeae is an ______ cocci
- Fusobacterium nucleatum is an _______ rod
- Bacteroides forsythus is an _________ rod
- Facultative
- aerobic
- anaerobic
- anaerobic
