Gavino 210 ICC – Flashcards
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| 3 Staining Methods |
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| Gram Staining Acid-Fast staining Endospore Staining |
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| Differential Staining |
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| -uses 2 or more stains to distinguish bacterial groups |
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| Simple staining |
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| uses one stain to see the morphology |
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| Morphology |
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| Shape, size and arrangement of cells |
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| Gram Staining steps and dyes usedWhat color is positive and negative? |
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| 1. Application of crystal violet (primary stain) 2. Application of Iodine (mordant) 3. Alcohol wash (decolorization) 4. Application of safranin (counterstain) -Purple= + -Pink= - |
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| Acid Fast Staining steps and dyes usedWhat color is positive and negative? |
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| 1. Application carbolfuchsin (primary stain) 2. Heat (mordant) 3. Acid Alcohol (decolorizer) 4. Methylene Blue (counterstain) -Pink = + -Blue = - |
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| Endospore Staining Steps and dyes usedWhat color is positive and negative? |
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| 1. Application of Malachite Green (primary stain) 2. Heat (Mordant) 3. Safranin (Counter stain) -Green = + -Pink = - |
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| What are some structural characteristics of Gram Positive cell walls? |
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| -think layer of peptidoglycan -Wall teichoic acid -Plasma membrane under layer of peptidoglycan -Tetrapeptide side chain -Peptide cross-bridge |
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| What are some structural characteristics of Gram Negative cell walls? |
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| -Thin layer of peptidoglycan -Outer layer of phospholipids -lipoproteins |
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| Diplococcus |
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| round cells arranged in twos |
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| Streptococcus |
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| round cells arranged in chains |
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| Staphylococcus |
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| round cells arranged in grape like clusters |
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| Diplobacillus |
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| rod cells arranged in twos |
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| Streptobacillus |
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| rod cells arranged in chains |
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| Ribosomes |
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| made of small 70S proteins and rRna; sites for protein synthesis |
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| Cytoplasm |
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| area inside the plasma membrane; contains 80% water |
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| Inclusions |
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| used to store materials such as lipids, polysaccharide, phosphate |
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| Glycocalyx |
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| slimy,sticky material made of polysaccharides and polypeptides; used by bacteria to attach onto surfaces (biofilm) |
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| Capsule |
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| hardened glycocalyx used by bacteria for protection against phagocytes |
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| Neucleoid |
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| made of DNA and contains most of the genes |
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| Plasmid |
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| small, circular, self replicating DNA that may contain a few genes |
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| Fimbriae |
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| many short hair like protein appendages used for attachment |
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| pilus |
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| hair like protein appendage used to transfer DNA during conjugation |
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| flagella |
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| protein filaments used for movement, have different arrangements |
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| axial filament |
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| protein fibrils that spiral around a cell used for movement |
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| Plasma membrane |
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| made of two layers of phospholipids and proteins that act as transporters, channels, receptors and enzymes |
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| 3 Functions of Bacterial Plasma Membrane |
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| 1. Selective permeability 2. Bacteria contains enzymes to make energy ATP (ex. electron transport chain in the plasma membrane) 3.Photosynthesis |
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| Selective permeability |
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| (semipermeable) allows certain molecules or ions to pass through the membrane; but others are prevented from passing through the membrane |
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| Chromophores and Thylakoids of plasma membrane |
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| contain pigments and enzymes for photosynthesis in photosynthetic bacteria |
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| Passive Processes |
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| do not require ATP because the substances move with their concentration gradient |
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| Concentration Gradient |
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| an area of high concentration to an area of low concentration |
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| Simple Diffusion |
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| substances move from an area of high concentration to an area of low concentration and do not require transporter proteins |
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| Facilitated Diffusion |
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| need transporter proteins to move substances from an area of high concentration to an area of low concentration |
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| Active Process |
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| Require ATP to move substances against their concentration gradient |
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| Active transport |
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| require transporter proteins and ATP to move substances against the concentration gradient |
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| Group Translocation |
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| substances are chemically altered during their transport through the membrane; need transporter proteins and ATP to move substances against the concentration gradient |
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| Solution |
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| Solute dissolved in solvent |
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| Solute |
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| a substance dissolved in another substance |
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| Solvent |
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| a dissolving medium |
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| Osmosis |
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| movement of water with its concentration gradient |
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| Osmotic pressure |
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| the force needed to stop the flow of water across the membrane |
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| Cell lysis |
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| occurs when the movement of water into the cell exceeds the ability of the plasma membrane to withstand the osmotic pressure |
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| True or False Fungi are sensitive to osmotic pressure |
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| False Bacteria are sensitive to osmotic pressure and fungi are resistant to osmotic pressure |
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| What substances can damage the plasma membrane of bacteria? |
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| Alcohol Anitbiotic (polymyxin) Detergent |
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| Why is the plasma membrane a good terget for antibiotic? |
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| Atp is produced in the plasma membrane and if that is destroyed then the cell cannot function |
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| Gram negative cell wall.... |
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| Has a outer membrane made of lipoprotein which is harder to stain |
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| Gram positive cell wall.... |
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| Has a thicker layer of peptidoglygan and holds the primary stain better Teichoic Acid |
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| Eukaryotic Animal cell Plasma membrane |
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| Phospholipids bilayer that controls transport of substances |
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| Eukaryotic Animal cell Lysosomes |
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| contains enzymes that can digest germs |
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| Eukaryotic Animal cell Ribosomes |
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| larger size involved in protein synthesis |
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| Eukaryotic Animal cell Nucleus |
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| contains DNA organized into chromosome pairs and histone proteins |
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| Eukaryotic Animal cell Rough Endoplasmic Reticulum |
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| membranous organelle involved in modifying proteins |
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| Eukaryote (Humans) Plasma Membrane Composition Function |
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| -Phospholipid bilayer, sterols -transports substances |
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| Eukaryote (Humans) Cell Wall Composition Function |
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| Absent |
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| Eukaryote (Humans) Ribosomes Composition Function |
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| -Larger size -protein synthesis |
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| Eukaryote (Humans) Chromosomes Composition Function |
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| -23 chromosome pairs (DNA) -genes control cell function |
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| Eukaryote (Humans) Membranous Organelles Composition Function |
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| Mitochondria, Endoplasmic reticulum and lysosomes |
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| Eukaryote (Humans) Cell Division Composition Function |
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| Mitosis and Meiosis |
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| Prokaryote (Bacteria) Plasma Membrane Composition Function |
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| -phospholipid bilayer, proteins -transports substances, energy production |
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| Prokaryote (Bacteria) Cell Wall Composition Function |
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| -peptidoglycan -maintains shape, protection |
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| Prokaryote (Bacteria) Ribosomes Composition Function |
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| -Smaller size -genes control cell function |
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| Prokaryote (Bacteria) Membranous Organelles Composition Function |
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| -Absent |
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| Prokaryote (Bacteria) Cell Division Composition Function |
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| Binary Fission |
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| Physical Growth Requirements |
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| 1. Temp 2. Oxygen 3. Chemical and Physical Requirements |
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| 4 Groups of microbes classified by temperature |
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| 1. Mesophiles 2. Psychrotrophs 3. Thermophiles 4. Hyperthermophile |
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| Mesophiles |
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| -moderate-temp loving, can grow from 10-48 degrees celcius ex. human pathogens like body temp (37) ex. grow in room temp (25) |
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| Psychrotrophs |
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| can grow from 0-30 degrees celcius ex bacteria that can spoil food in the refrigerator (5) |
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| Thermophiles |
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| heat loving, can grow from 40-70 degrees celcius |
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| Hyperthermophile |
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| hot temperature loving, can grow from 65-110 degrees celcius ex. Archaea |
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| 5 groups of microbes classified by the amounts of o2 needed |
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| 1. Facultate Anaerobes 2. Obligate Aerobes 3. Microaerophiles 4. Obligate Anaerobes 5. Aerotolerant Anaerobes |
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| Facultate Anaerobes |
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| Prefer O2 but can survive without it ex. E. coli |
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| Obligate Aerobes |
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| require a lot of O2 |
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| Microaerophiles |
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| require very little O2 |
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| Obligate Anaerobes |
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| killed by O2 |
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| Aerotolerant Anaerobes |
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| Don't use O2 but can live with O2 |
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| Organic Molecules |
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| Carbohydrates, proteins, lipids, nucleic acids |
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| Nucleic acids |
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| DNA, RNA, ATP |
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| Mineral elements |
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| iron, copper, zinc |
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| 7 Chemical and Physical Requirements of microbes |
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| 1. Carbon 2. Nitrogen 3. Sulfur 4. Phosphorus 5. Trace elements 6. Organic Growth Factors 7. pH |
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| Carbon |
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| microbes need to make all kinds of organic molecules to form their cell structures, carbon is used as a source of energy |
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| Nitrogen |
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| needed to make the organic molecules proteins and nucleic acids ex. most is acquired from protein sources (food) ex. nitrogen fixing bacteria get it from the atmosphere |
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| Sulfur |
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| needed to make amino acids and vitamins |
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| Phosphorus |
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| needed for microbes to make nucleic acids and the bilayer of the plasma membrane |
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| Trace Elements |
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| mineral elements needed in small amounts for making cofactors of enzymes |
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| Organic Growth Factors |
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| essential organic compounds from the environment ex. vitamins, amino acids, purines and pyrimidines |
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| Purines |
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| A and G Bases of DNA and RNA |
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| Pyrimidines |
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| T, C, U Bases of DNA and RNA |
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| pH for Bacteria |
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| Like neutral 6.5-7.5 pH |
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| pH for Fungi |
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| molds and yeasts like acidic 5-6 pH |
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| Culture medium |
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| nutrients prepared to grow microbes ex. nutrient agar |
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| Sterile |
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| contains no living microbes |
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| Inoculum |
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| microbes introduced into a culture medium to initiate growth |
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| Culture |
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| microbes growing in or on culture medium |
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| Pure culture |
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| contains only one species or strain |
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| Colony |
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| a visible mass of microbial cells that arose from a single cell, spore, or a group of attached cells |
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| Preserving Bacterial Cultures Short term |
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| Slows down metabolism |
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| Preserving Bacterial Cultures Long Term |
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| Totally stops metabolism for years |
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| Deep-freezing |
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| Long term bacterial preservation -in liquid nitrogen -50 to -95 decrees celcius |
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| Lyophilization |
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| Long term bacterial preservation -freeze drying dehydrate in a vacuum then freeze -54 to - 72 degrees celcius |
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| 2 ways to obtain pure cultures |
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| 1. Pour Plate Method 2. Streak Plate Method |
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| Complex media |
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| chemical composition varies slightly per batch |
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| Chemically Defined Media |
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| the exact chemical composition is known |
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| Selective Media |
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| suppress the growth of unwanted microbes and encourage the growth of the desired microbe ex. sabourand agar has acidic pH5 for fungi ex Mannitol Salt Agar for Staphylococcus aureus |
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| Differential Media |
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| make it easier to distinguish colonies of desired microbe ex. hemolytioc bacteria form halo on Blood agar and EMB agar |
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| Enrichment media |
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| encourage the growth of a desired microbe ex enrichment broth for staphylococcus |
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| Bacterial Growth Curve |
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| 1.lag phase 2.log phase 3.stationary phase 4.Death phase (log arithmic decline) |
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| lag phase |
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| no cell dvision |
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| log phase |
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| Exponential growth, optimum conditions |
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| Sationary phase |
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| slower growth, new cells are produced at the same rate cells are dying |
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| Death phase |
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| dying cells exceed new cell production |
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| Measuring Microbial Growth 4 Direct Methods |
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| Plate count Most Probable Number (MPN) Direct Microscopic Count Filtration |
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| Measuring Microbial Growth 3 indirect methods |
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| Turbidity Metabolic Activity Dry Weight |
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| Plate Count Advantage Disadvantage |
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| a common method for estimating the number of bacteria in a sample, assumes that each bacterial cell grows into a single colony -counts only the number of living cells -need a lot of materials to perform serial dilutions of a sample -requires time for incubation (Direct Method) |
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| CFU |
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| colony forming units |
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| Most Probable Number (MPN) |
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| a statistical estimation of bacterial count, this method is used when microbes won't grow on solid media (Direct) |
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| Liquid differential media |
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| uses lactose for broth for coliforms for microbes that only grow in liquid |
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| Direct Microscopic Count Advantage Disadvantage |
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| uses a special slide that can hold a measured volume of bacterial suspension -a fast way of counting the number of cells -requires a high concentration of cells in a sample -does not work if the bacteria are motile -counts living and dead cells (direct) |
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| Filtration |
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| bacteria are first sieved out onto the surface of a membrane filter then transferred to a culture medium and the CFUs are counted a useful method when the bacterial count is low (direct) |
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| Turbidity |
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| measures the level of cloudiness uses a spectrophotometer to measure the amount of light that passes through a suspension of cells (indirect) |
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| Metabolic Activity |
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| measures the amount of metabolic products such as acids gas and enzyme production (indirect) |
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| Dry weight |
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| used for filamentous organisms (fungi and algae) that can be grown, dried and weighed (indirect) |
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| 4 groups of organic molecules |
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| Carbon, hydrogen, oxygen, and nitrogen |
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| atoms found in carbohydrates |
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| Carbon, hydrogen, oxygen 2:1 hydrogen to oxygen |
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| atoms found in lipids |
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| Carbon, Hydrogen and Oxygen |
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| monosaccharides |
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| simple sugars 3-7 carbon molecules |
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| disaccharides |
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| monosaccharides bonded in dehydration synthesis reaction and can be broken down by hydrolysis |
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| polysaccharides |
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| 10s or 100s of monosaccharides joined through dehydration synthesis -not soluble in water |
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| simple lipids |
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| fats, triglycerides contain glycerol and fatty acids |
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| complex lipids |
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| contain phosphorus, nitrogen, sulfur, carbon, hydrogen and oxygen -glycerol and 2 fatty acid chains and a phospholipid group |
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| atoms found in proteins |
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| Carbon Hydrogen Oxygen and Nitrogen |
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| Steroids |
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| 4 interconnected Carbon rings |
