Micro Biology HU Spring 2014: TEST 1

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Micro Biology Test Preparation/Pathogen Knowledge

Spirochete

Staphylococcus

Streptococci

Carl Richard Woese

Famous for defining the Archaea (a new domain or kingdom of life). Created the 3 Domain system based on RNA structure.

Carl Linnaeus

Swedish Biologist who developed the 5 Kingdom System of Classification.

Staphylococcus aureus

Microorganism responsible for Staph Infection.

Bacteriology

Study of Bacteria

Mycology

Study of Fungi

Phycology

Study of Algae

Archaea

Domain which includes extremophiles.

Virology

Study of Viruses

Halophiles

Microorganisms which thrive in extremely "salty" environments.

Acellular

Term which means: "Not composed of cells."

Corona Virus

Microorganism (virus) which causes MERS.

Algae

Photosynthetic Eukaryotes.

Protozoology

Study of Protozoans.

Helminths

A polyphyletic group of eukaryotic parasites (flat worms and round worms).

Arthropods

An invertebrate animal having an exoskeleton (external skeleton), a segmented body, and jointed appendages (these include fleas, ticks, and mosquitos).

Epidemiology

A study of the spread and prevalence of disease.

Etiology

A study of the cause of disease.

Black Death

(1347-1665) Plague that killed off 1/3 of Europe's population.

Miasma

a foul, poisonous vapor.

Louis Pasteur

Father of Modern Medicine

Francesco Redi

Italian scientist who first attempted to disprove the idea of Abiogensis with his meat experiment.

Edward Jenner

English doctor who vaccinated people against small pox (with the help of the milk maids).

Louis Pasteur

Proposed Germ Theory.

Louis Pasteur

Worked with Fowl Cholera. Vaccinated Chickens by injecting aged cultures.

Antony van Leeuwenhoek

Cloth merchant who discovered "animalcules."

Louis Pasteur

Proved Fermentation due to yeast; developed Pasteurization; and disproved Abiogensis.

Louis Pasteur

Saved a boy from rabies by using dried rabid rabbit spinal cords.

Robert Koch

The Father of Medical Microbiology. Proved Germ Theory through the development of pure culture techniques.

Koch's Postulates

Method of thinking used to associate an organism with a disease.

Koch's Postulates ( All 4)

1. The organism must be found in all cases of the disease.
2. The organism should be isolated in a pure culture.
3. When the pure culture of the organism is injected into a susceptible host, the identified disease should result.
4. The identical microbe should isolated from this host and isolated in a pure culture.

Robert Koch

Was able to isolate Microbacterium Tuberculosis. Studied Cholera, Plague, and African Sleeping Sickness. Identified and discovered 20 major bacterial pathogens.

Agar

Seaweed extract.

40 Angstroms = ? (millimeters)

.000000004 milimeters

milimeter (10^?)

10^-3

micrometer (10^?)

10^-6

nanometer (10^?)

10^-9

angstrom (10^?)

10^-10

Resolution

the ability to separate two items as separate and distant entities.

Light Microscope

1000x-2000x

Max Magnification for a Light Microscope

Light Microscope

Uses visible light. Sometimes called a "bright field microscope."

Resolution Limit for a Light Microscope (____um)

.2

Compound Light Microscope

Uses visible Light, has a modified condenser which directs light towards the side of the specimen, is sometimes called Dark Field Microscope, and illuminated the specimen while leaving the surrounding dark. It typically used for bacteria which cannot be stained.

Phase Contrast Microscope

Uses visible light, takes advantage in refractive indexes (optical density) of cellular structures, to see objects light waves must be "in phase." Typically used for unstained specimens.

Differential Interference Microscope

Similar to phase contrast, contains additional light source and multiple prisms, 3-D appearance and more colorful. Also referred to as a "Nomarski Scope."

Fluorescent Scope

Modified microscope with an ultraviolet light source and protective eye filters uses dye that emit visible light when bombarded with shorter UV rays (useful in diagnosing infections using clinical specimens and fluorescent labeled antibodies.

Immunofluorescence

Fluorescent-antibody techniques. Antibodies are attached to dyes, and if the antibody binds to its specific target, the target cell will show via fluorescence microscopy.

Confocal Microscope

Laser illuminated, a laser scans over the specimen with the help of some scanning mirrors and the image is put on a computer for analysis, lasers are used because of their wavelength, salt can be seen in this microscope

Electron Microscope

Microscope that forms an image by focusing beams of electrons onto a specimen.

Scanning Electron Microscope

A microscope that is especially useful for the detailed study of the surface of a specimen; electron beams scan the surface of the sample, which is usually coated with a thin film of gold, allowing electrons on the surface to be deleted and translated into an image; result is a 3D topographic image. Can magnify from 1000x-100,000x

Transmission Electron Microscope

Magnifies about 1 million times; used to view the inside of the cell; a disadvantage is that the specimen must be: dead, frozen and cut into small slices in order to view it. Can magnify from 10,000x-1,000,000x

Eukaryotes

Nucleus: Present
Membrane Bound Organelles: Present
DNA Structure: Multiple chromosomes. Histons Present
Chlorophyll: When present, in chloroplasts.
Ribosomes: In Cytoplasm and Rough ER, larger than Prokaryotes
Reproduction: Mitosis
Cell Walls: Only in Plants and Fungi.

Prokaryotes

Nucleus: No Nucleus.
Membrane Bound Organelles: None
DNA Structure: Single closed loop. No histons.
Chlorophyll: When present, dissolved in cytoplasm.
Ribosomes: In the Cytoplasm, Smaller than Eukaryotes.
Reproduction: Binary Fission
Cell Walls:Usually Chemically Complex

Thiomargarita namibiensis

Organism believed to be responsible for cycling sulfur. (Macroscopic)

Helicobacter pylori

Bacteria responsible for causing ulcers.

Pleomorphic

Taking on many shapes.

Broth culture

Type of culture best for determining bacterial morphology/

Antigen

A substance that induces a state of sensitivity or immune responsiveness (immunity).

Epitope

a localized region on the surface of an antigen that is chemically recognized by antibodies; also called antigenic determinant

Innate Immunity

Non-specific
Non-inducible
No "memory" produced
Recognition of conserved regions

Acquired Immunity

Specific
Inducible
Memory produced
Recognition of conserved and non-conserved regions.

B Cells

Cells manufactured in the bone marrow that create antibodies for isolating and destroying invading bacteria and viruses.

T Cells

Specialized white blood cells that fight disease either by activating B cells (helper____cells) or by attacking antigens directly (killer____cells)

Helper T Cell marker(s)

CD 4

Cytotoxic T Cell marker(s)

CD 8

Regulatory T Cell marker(s)

CD 25+ and CD 4

Serology

The "in vitro" study (in glass) of interactions between antibodies and anitgens.

The 3 Domains

Archaea, Bacteria, and Eukaryote

Fimbriae

Name the structure exhibited by these cells.

Freeze Etching

Using a cryogenic knife to cut into a cell, and exposing the nuclear membrane.

Cell Mediated Immunity (T Cell)

Type of acquired immunity mediated by Helper T Cells and Cytotoxic T Cells

Shadow Casting

Spraying a cell surface with a shower of heavy metals at a 45 degree angle.

Humoral Immunity (B Cell)

Type of acquired immunity mediated by B Cells.

For bacteria, this means run

Clockwise

For bacterial, this means tumble

Counter-clockwise

Chemotaxis

moving towards (positive) or away (negative) from a chemical stimulus

Phototaxis

moving towards (positive) or away (negative) from a light stimulus

Toll-like Receptors (TLRs)

Receptors on phagocytes that often bind bacterial appendages, signaling as a trigger for the immune system.

Flagellin

Protein which composes the flagellum

Endo-flagellum

Flagellum used by bacterial spirochetes, not a true appendage, offers crude motility

Pilus

filamentous projection on a bacterial cell, used not for motility but for adhering to other bacterial cell (especially for mating) or to animal cells. Composed of "pilin" protein. Similar to "fuzz on a peach"

Atrichous

Non-motile (not mobile)

Monotrichous

1 flagellum

Amphitrichous

2 Flagellum, one on each end of the organism

Lophotrichous

Multiple flagellum on one side

Peritrichous

Multiple flagellum, surrounding the bacterial cell

The 3 parts composing the Flagella structure

The filament, the hook, and the basal body (the motor)

Conjugation

Transfer of DNA from one cell to another

Tissue Tropism

The ability of microorganisms to associate with certain tissues of the body

Virulence Factor

Any microbial factor that can promote the onset of disease

Fimbriae

Functions in attachments to substrates (help things bind to rocks in some cases), important virulence factor, composed of protein similar to pilin, and is tissue specfic

Three parts of the cell envelope

1. Glycocalyx
2. Cell Wall
3. Cell Membrane

Glcocalyx

Outer most layer of cell envelope. Two distinct regions: Slime Layer and Capsule

Slime Layer

Composed of polysaccharides, function in retention of water and cell nutrients, can be easily removed in a stream/running water

Capsule

Composed of repeating ogliosaccharide subunits or proteins, functions in the prevention of phagocytosis, cause adherence and promotion of biofilm formation

Stages of Phagocytosis

1. Endocytosis
2. Phagosome
3. Phagolysosome
4. Late-endosome
5. Exocytosis

Biofilm

A community of cells surrounded by a capsule, composed of organic and inorganic constituents, form on surfaces in aquatic systems, resistant to biocides, release of organisms into bulk fluid

Stages of Biofilm formation

1. Association
2. Adhesion
3. Microcolony Formation
4. Biofilm Formation

Biofilm Resistence to Antimicrobial Agents Steps

1. Slow penetration
2. Resistant Physiology
3. Adaptation

Impacts of Biofilms

Teeth, cooling water (Nuclear reactors), food processing, ship hulls, oil recovery, drinking water, paper manufacturing, and medical implants

Cell Wall Function

Binds cations (Mg++ and Ca++), rigidity and shape of the cell, protection from osmotic lysis, active site of certain antimicrobial agents (penicillin), antigens, and virulence factors

Peptidoglycan Layer

Composed of peptides and sugars, NAM-NAG-NAM-NAG-NAM alternating chains, connected via Beta 1-->4 linkages (Lysozymes can break this linkage)

NAM

N-acetyl-muramic acid

NAG

N-acetyl-glucosamine

Cell walls of Gram Positive Bacteria

Composed of:
1. Peptidoglycan Layers
2. Teichoic Acids

Teichoic Acids

1. Wall teichoic acids
2. Lipoteichoic acids

Wall teichoic acids

Teichoic acids within the cell wall

Lipoteichoic acids

Teichoic acids within the cell membrane

Teichoic Acids

Teichoic acids, a fortification wall, are bacterial polysaccharides of glycerol phosphate or ribitol phosphate linked via phosphodiester bonds. Play an important role in cell growth and antigenicity

Gram Positive Cells

Important antigens within the gram + cells

Lysozyme

an enzyme found in saliva and sweat and tears that destroys the cell walls of certain bacteria

Peptidoglycan Structure:

Alternating L and D amino acids in sets of 4 (Linked only to the NAMs, not the NAGs), Pentaglycine (5 glycines) connects the linkages of NAG-NAM chains

Gram Positive Linkages present in peptidoglycan layer?

Yes. ~100% Cross-linking. In Gram + organisms, you can have ~40 layers of peptidoglycan, all with cross linking

Gram Negative Linkages present in peptidoglycan layer?

Yes. ~20% Cross-linking. In Gram - organisms, you can have ~1-3 layers, with only ~20% cross linking

Porins

Proteins that allow the passage of certain ions and small polar molecules through membranes.

Periplasmic Space

In Gram-negative cells, the space between the cell membrane and the outer membrane containing peptidoglycan and periplasm.

Lipopolysaccharide (LPS)

Provides a protective barrier; help in adhesion, B Cell and Macrophage activator. The longer the sugar residue, the more antigenic and mucoid it becomes. It is composed of 3 sections: O Antigen region, Core polysaccharide region, and the Lipid A regions. Lipid A region is endotoxic (Causes fever, shock, spontaneous abortion, and death)

Essay Question for the first exam:

In detail, give the structure of the Gram Positive and Gram Negative cell walls. (At least a paragraph for each)

Purple color =

Gram Positive

Red color =

Gram Negative

Mordant

Means the dye forms a complex with crystal violet.

Exceptions to GP and GN structure:

1. Mycobacterium and Nocardia
2. Mycoplasma
3. Archaea
4. Rickettsia and Chlamydia

Mycobacterium produce______?

Mycolic Acid

Mycobacterium and Nocardia

Both genera cause respiratory disease, contain peptidoglycan along with high concentrations of lipids - mycolic acid, resist decolorization in the presence of acid. Therefore, these bacteria are termed Acid Fast Bacilli (AFB)

Acid Fast Stain

Heat forced the dye in. AFB- = Blue color; AFB+ = Red color

MDR

Multi-drug Resistant

XDR

Extensively Drug Resistant

Mycobacterium Avium-intracellulare Complex

A type of infection particularly common in HIV patients

Mycobacterium Leprae

Bacteria which causes leprosy

Classic drugs with which to treat TB:

Isoiazid and Rifampin

Mycoplasma

Are found in humans and most have no cell wall. Can cause pneumonia. Not affected by antibiotics which target the cell wall, no morphology

Rickettsia

Obligate Intracellular parasite, cannot be grow outside of the cell (but IS NOT a virus), transmitted by arthropods, (Rocky Mountain Spotted Fever and Typhus)

Chlamydia

Obligate Intracellular parasite, Chlamydia trachomatis is the most often seen, most common venerial disease in the world, the most common cause of preventable blindness in the world today

Cell Membrane

Composition: Phosopholip bilayer with embedded protein
Function: Osmotic barrier, transport, energy extraction, nutrient catalysis and processing, enzymatic biosynthesis and degradation

Protoplasm

DNA
Bacterial Cytoskeleton
Ribosomes
Inclusions
Endospores

Nucleoid Region

DNA containing region. No membrane around the nucleoid region. Super-coiled DNA.

Plasmids

Extra chromosomal DNA elements. Can replicate independent of chromosome. Sometimes aid organisms by giving them a selective advantage (ex: antibacterial agent resistance, metal resistance, virulence factors).

Resistance Plasmids

R Plasmids

Fertility Plasmids

F Plasmids

Bacterial Cytoskeleton

Chemically similar to tubulin and actin. May help stabilize structure of bacterial cell and play a role in movement.

Ribosome

Small particles composed of rRNA and protein which direct the synthesis of proteins during translation. Composed of large and small subunits. Not bound to membrances. Smaller in prokaryotes than in eukaryotes.

Velocity Sedimentation

How fast or slow a ribosome moves through a sucrose gradient. Dependent on mass and surface area.

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