Rod shaped- Bacilli

Sphere shaped- Cocci

Yeast- Unicellular circles/ovals

Solid:

• General Purpose; TSA plate
• Enriched Media; TSA plate with 5% sheep's blood
• Differential Media; MSA plate
• Selective Media; EMB plate

Semi-soild:

More like pudding

Liquid:

Nutrient Broth Tube

• Every time you open the specimen tube; the top of the tube must be sterilized
• Sterilize the inoculating loop before & after each use.

• Used for observing shape & arrangement of cells
• One stain used

Direct Stains:

Stains cells

Negative Stains:

Stains background & leaves part of or all of the cells unstained.

Glycocalyx

• Makes the cell sticky, helping them attach better to the surfaces
• Evade host immune system

example: Streptococcus pneumoniae or  Mycobacterium

Gram stain

Endospore stain

Acid-fast stain

Differentiates the two major groups of bacteria by the cell wall characteristics. 

Gram + stains purple, Gram - stains red/pink

• Make smear (placing bacteria on slide, let it dry,  then heat fix)
• Cover smear with Crystal Violet; rinse off with water
• Cover smear with Iodine; rinse off with water
• Cover smear with Ethyl Alcohol; rinse off with water
• Cover smear with Safranin; rinse off with water
• Blot Dry

Primary - Crystal Violet

Mordant - Iodine

Decolorization - Ethyl Alcohol

Counter-stain - Safranin

Clostridium & bacillus produce endospores

• Endospores: A survival structure that allows the bacterial cell to survive unfavorable conditions
• Under favorable conditions they germinate & release a viable vegetative cell

• Can differentiate between acid-fast and non acid-fast bacteria 
• Acid-fast have mycolic acid in cell walls making them waxy

Eukaryotic:

• Unicellular (yeast)
• Multi-cellular (molds)

• Beta (β): Complete breakdown of RBCs (clearing around colonies)
• Alpha (α): Partial destruction of RBCs (greenish brown color around colonies)
• Gamma (γ): No lysis of RBCs (intact Blood agar around colonies)

The study of living organisms that are generally too small to be seen by the naked eye. 

Micro; small

Bio; life

logy; study of

• Bacteria
• Viruses
• Parasites
• Fungus

• Soil
• Water
• Surface areas
• People & animals

• Resident flora
• Bacteria and other microbes
• Initial colonization of newborn; birth canal is the start up

• Intestines
• Skin
• Eyelashes
• Nails
• Nose hairs

• First simple microscope (1667)
• "Aminacules"; tiny organisms
• Discovered 3 bacterial shapes' bacilli, cocci, spirals
• Examined pond water, wells, lakes, rivers, blood, urine, feces, & semen

• Observed jars of raw meat for signs of "worms"
• Conclusion: Decaying meat does NOT give rise to maggots

• Experimented with Spontaneous Generation

• Used swan-necked flasks to disprove spontaneous generation; Theory of Biogenesis
• Founder of Microbiology
• Germ Theory of Fermentation
• Pasteurization; heating to 50-60 ºC
• Germ theory of Disease
• Vaccines

• First to isolate anthrax in sheep
• Developed Koch's Postulates; technique of isolating & identifying specific pathogens that cause specific diseases, serious of proofs that verified the germ theory.

- Isolate the suspected agent from the victim

-Grow the agent in pure culture

-Infect healthy host with the agent

-Isolate the "same" organism from the new victim

• Used Carbolic acid to reduce wound infections by 2/3
• Introduced what is now known as the aseptic technique
• Where "Listerine" came from

• Father of Immunology
• 1798
• Milkmaids; Contracted smallpox, Immune to smallpox

• Advocate of cleanliness in military hospitals
• Kept detailed records to support her theory that unsanitary conditions led to deaths

• Word laid the foundation for; Epidemiology & infection control
• Mapped the occurrence of cholera cases in London; Linked cholera to a contaminated well

• Developed the most commonly used staining technique in microbiology; Gram stain

Prokaryotic:

• Small
• Lack a nucleus & organelles

Eukaryotic:

• Larger
• Have nucleus & organelles

• A system for organizing, classifying, & naming living things
• Primary concerns of taxonomy are classification, nomenclature, & identication

• Domain
• Kingdom
• Phylum
• Class
• Order
• Family
• Genus
• Species

• Coccus
• Rod or Bacillus
• Curved- vibrio
• Spiral- Spirillum
• Corkscrew- Spirochete
• Pleomorphic

• Single 
• Pair (diplo)
• Chains
• Clusters

• Mycelium
• Vegetative hyphae
• Spores
• Budding

• Tinea Captis (head)
• Tinea Barbae (beard)
• Tinea Corporis (body); ringworm
• Tinea Cruris (groin)
• Tinea Pedis (feet)
• Tinea Manuum (hands)
• Tinea Unguium (Nails)

• Tetanus
• Lockjaw
• Neuromuscular Disease

• Increase in population of microbes; increase in #

• The division of bacterial cell
• Parental cell enlarges & duplicates its DNA
• Septum formation divides the cell into two separate chambers
• Complete division results in two identical cells

• Generation Time: the time required for a complete division cycle (doubling)
• Growth Rate: number of generation per hour.

Note: E. coli generation takes 20; 3 generations per hour

• Lag phase
• Log phase (exponential growth phase)
• Stationary phase
• Death phase

• Spieces
• Available nutrients
• Temp
• pH
• Gas requirements
• Toxic concentrations (salt)
• Competition with other microbes

• Source of energy: chemical; breakdown of organic & inorganic molecules-methane, light; photosynthesis
• Source of matter for building additional cells: CHNOPS & trace elements

1. Inoculation
2. Incubation
3. Isolation
4. Identification

1. Culture sample
2. Colony morphology
3. Gram Stain
4. Biochemical tests

Pure: one bacteria

Mixed: two or more bacteria

Contaminated: something is there that isn't suppose to be there

E. coli: Metallic Green

Enterbacterial: pink/purple

Positive: maroon/purple

Negative: Cloudy/clear

It determines if the test has the right enzyme; Indophenol; to oxidase the presence of oxygen

Positive: layer of red

Negative: no red layer

Positive tests indicate the production of indole by bacteria

Positive: Bubbles; Staph

Negative: No bubbles; Strep

Positive:

Staphylococcus aureus

Staphylococcus epidermidis

Micrococcus luteus

Negative:

Streptococcus mutans

Differentiate between micro & micro-nutrients

Micro (trace elements): present in smaller amounts for enzyme synthesis & maintenance of protein structure (Zn, Mn, Cu, & Fe)

Macro: Required in large amounts for cell structure & metabolism (C, H, & O)

Growth factors: organic nutrient that must be provided to the bacteria

Examples: amino acids & vitamins

• Psychrophiles: -5 to 20 ºC
• Mesophiles: 15 to 45 ºC
• Thermophiles: 45 to 80 ºC
• Hyperthermophiles: 70 to 105 ºC

Growth is called exponential because it's written like a geometric exponent. In 20 generations there are 1,048,576 cells. 

Lag phase: Very little to no bacterial growth

Log (exponential) phase: Bacterial cells doubling at a constant, exponential rate

Stationary phase: The number of viable bacteria stays the same

Death (decline) phase: The bacteria dies

Catabolism: Breakdown of complex organic molecules into simpler compounds, releases energy

Anabolism: The building of complex organic molecules from simpler ones, requires energy

Reduction: The gain of hydrogen &/or electrons

Oxidation: The loss of hydrogen &/or electrons

• Anaerobic
• Aerobic → Krebs cycle
• Anaerobic fermentation

Signs: Things everyone can see

Symptoms: Things patients precieve

Genotype: Set of genes in the genome

Phenotype: Physical features & functional traits of the organism, can change based on which genes are expressed ("on")

Triplet cod that specifies a given amino acid. Multiple codes for amino acid. 20 Amino acids.

Start codon. Stop codon. 

RNA polymerase: Search DNA template for promoters

DNA pol I: Removes the RNA primers & replaces them with DNA

DNA pol III: Recognizes the primers & binds to the 3' end.

DNA helicase: Separates strands of a double helix or a self-annealed RNA molecule using the energy from ATPhydrolysis

DNA ligase: Seals the gaps between fragments on the lagging strand.

Adenine, Guanine, Cytosine, & Thymine. A&T, G&C.

Purines: Adenine & Guanine

Pyrmindines: Thymine & Cytosine.