Chapter 7 Microbial Growth Flashcards

1
Q

Macronutrients

A

(C,H,O,P,N,S)
Required in large amounts
Metabolism and cell structure
K+,Ca++,Mg++,Fe+++ moderate quantities

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2
Q

Micronutrients

A

(Mn,Zn,Ni,Co,Cu,Mo)
Small amounts - difficult to determine exact requirements (tapwater sufficient)
usually enzymatic function

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3
Q

Chemical requirements for growth

A
  1. Carbon - CO2 or organic
  2. N, S, P - DNA & RNA, protein, some lipids
  3. Trace Elements - Metal ions - only certain cells and in tiny small amounts
  4. Growth Factors - not made by organism
    •purines and pyrimidines: required for synthesis of nucleic acids (DNA and RNA)
    •essential amino acids: required for the synthesis of proteins
    •vitamins: needed as coenzymes and functional groups of certain enzymes
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4
Q

Media formulations

A

Chemically defined media - exact amounts of pure chemicals (research)
Complex media - composition varies (enzymatic)

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5
Q

Specialty medias

A

Selective - inhibit growth of certain species.
Differential - most things will grow- but will look different
Selective and differential - utilizes both aspect

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6
Q

Photoautotroph - Oxygenic

A

Produces oxygen (O2)
(Inorganic)CO2 + H2O + light = (CH2O)n + O2
cyanobacteria, algae, green plants

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7
Q

Photoautotroph - Anoxygenic

A

Does NOT produce oxygen
use inorganic material like hydrogen sulfide (to sulfur) or hydrogen gas (to water)
are obligate anaerobes (O2 toxic)
bacteriochlorophylls - green and purple sulfur bacteria

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8
Q

Photoheterotroph

A

Use light for energy
Use organic compounds like alcohols and fatty acids for carbon source
obligate anaerobes
Green and purple non-sulfur bacteria
Found in bogs, moist soil, paddy fields (light but no O2)

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9
Q

Chemoautotroph

A

Electrons from reduced inorganic compounds used for energy
CO2 used for carbon
remove electrons from H2, H2S, S, or Fe and combine them with CO2, O and H
deep-sea vent bacteria; hot springs

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10
Q

Chemoheterotrophs

A

Use organic molecules for both energy and carbon
Aerobic: uses oxygen as final electron acceptor
Anaerobic: uses sulfate, nitrate, etc. as final electron acceptor
most bacteria, all fungi, all protozoa and all animals, pathogens

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11
Q

Inorganoheterotrophs

A

Use reduced inorganic molecules for energy
Organic molecules for carbon source
some bacteria

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12
Q

Parasites and Saprobes

A
Parasites: derive nutrients from host
•Pathogens
•Some are obligate parasites
Saprobes: free-living microorganisms that feed on organic waste from dead organisms 
•Opportunistic pathogen
•Facultative parasite
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13
Q

Gas Requirements

A

Oxygen: O2
•Powerful oxidizing agent
•Must be “detoxified”

Carbon dioxide: CO2
•Essential for autotrophs

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14
Q

Oxygen Radicals (Must be neautralized even if organism doesnt use oxygen)

A

Toxic by-products of O2 use and exposure:
•Superoxide O2- superoxide dismutase
•H2O2
catalase, peroxidase
•Singlet Oxygen
photosynthetic organisms have carotenoids which scavenge the singlet oxygen and render it nontoxic

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15
Q

Oxygen Requirements- Obligates

A

Aerobes-
require O2 for growth (21% O2 in air)
they use O2 as a final electron acceptor in aerobic respiration
Anaerobes-
do not need or use O2 (toxic)
anaerobic respiration, anoxygenic bacterial photosynthesis, methanogenesis

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16
Q

Oxygen requirements- Others - Facultative anaerobes

A

Facultative anaerobes- Can switch between aerobic and anaerobic types of metabolism (O2 prefered, fermentation or ana-respiration if not)

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17
Q

Capnophiles

A

require a high level of CO2 and are grown in candle jars

18
Q

Oxygen requirements- Others - Aerotolerant anaerobes

A

Exclusively anaerobic type of metabolism Insensitive to the presence of O2
Produces superoxide dismutase and peroxidase enzymes
Ex: Streptococci

19
Q

Oxygen requirements- Others - Microaerophiles

A

Are aerobic, but need low concentrations of O2
<15% O2
Ex: Helicobacter pylori

20
Q

Anaerobic growth media:Fluid Thioglycollate Medium

A

Anaerobes are killed by exposure to oxygen
Use a reducing medium to deplete available oxygen
Heating the media also drives off oxygen

21
Q

pH Requirements

A

Neutrophiles: Most bacteria grow between pH 6.5 to 7.5
Acidophiles: grow below pH 4.0
Lactobacillus acidophilus (yogurt, ~pH 4)
Some molds and bacteria can tolerate acid (food spoilage)
Alkalinophiles: grow best under basic conditions (not many)

22
Q

Transport: Movement of Chemicals Across the Cell Membrane - Passive transport

A

does not require energy
substances exist in a gradient and move from areas of higher concentration toward areas of lower concentration
Types:
•Simple diffusion – small things slip between the phospholipids (CO2, O2, N2, water)
•Osmosis – diffusion of water
•Facilitated diffusion – requires a carrier

23
Q

Transport: Movement of Chemicals Across the Cell Membrane - Active transport

A

requires energy and carrier proteins
gradient independent
Types:
•Active transport
•Group translocation – transported molecule chemically altered
•Bulk transport – endocytosis, exocytosis, pinocytosis

24
Q

Diffusion

A

Net Movement of Molecules Down Their Concentration Gradient Passive Transport
No addition energy required

25
Osmotic Solutions
Isotonic - w or w/o cell wall equilibrium Hypotonic Cell wall - wall prevents bursting w/o cell wall - swelling then osmolysis Hypertonic cell wall - membrane shrinks inside wall -plasmolysis w/o cell wall - shrinking and distortion
26
Facilitated Diffusion
Monomer sized molecules need a transport protein to cross the membrane •Carrier protein •No additional input of energy required
27
Endocytosis
Bringing substances into the cell through a vesicle or phagosome •Phagocytosis ingests substances or cells •Pinocytosis ingests liquids
28
Symbiotic Relationships
mutualism – obligatory, dependent; both members benefit commensalism – commensal member benefits, other member not harmed parasitic – parasite is dependent and benefits; host is harmed
29
Quorum sensing
a group of microorganisms coordinate functions (to release digestive enzyme, toxins, transfer DNA, etc.) Quorum: critical number of cells Inducer molecules: coordinate some response
30
Counting Bacterial Growth
2n, where n is the number of generations exponential growth 5-10-20-40-80-160-320
31
Phases of Growth: Lag
no or slow growth, yet •culture “waking up”- synthesizing DNA, enzymes, ribosomes, etc. •or culture so dilute it is below limit of detection
32
Phases of Growth: Log
* Cell division as fast as possible (“balanced growth”) * Adequate nutrients and favorable environment * Appears linear on a log scale * Cells are dividing and growing in geometric progression (2,4,8,16,32,64,128,etc.) * Phase used to calculate generation time
33
Phases of Growth: Stationary
* Growth slows such that new cells = dying cells * Growth is limited by space, supply of nutrients * Depleted nutrients, and waste is accumulating
34
Phases of Growth: Death
* Number of cells dying increases exponentially due to build up of waste products. * Some remain viable
35
Direct Measures of Growth - plate counts
Plate Counts •enumeration of bacteria: Assumption that every live bacterium will produce a colony on a plate •Viable counts
36
Direct Measures of Growth - filtration
* Used when quantity of bacteria is low * Need to concentrate bacteria onto filter * Transfer bacteria from the filter to an agar dish * Touch the filter to an agar dish, then incubate to grow colonies
37
Direct Measures of Growth - Direct Cell Counts
* Petroff-Hausser cell counter * Use very accurate counting chamber under microscope (like a hemocytometer) * Use grid to establish number of organisms
38
Direct Measures of Growth - Automated Direct Counts
``` Coulter Counter •Separate based on charge •Automatic •Expensive •Breaks easily ``` Flow cytometry•Can count cells based on fluorescent labels•Separates live from dead•Must be able to manipulate organisms DNA
39
Direct Measures of Growth - Automated Direct Counts
``` Coulter Counter •Separate based on charge •Automatic •Expensive •Breaks easily ``` Flow cytometry •Can count cells based on fluorescent labels •Separates live from dead •Must be able to manipulate organisms DNA
40
Indirect Measures of Bacteria
Turbidity: Use spectrophotometer to measure amount of light scattered by a culture Metabolic activity: measure amount of product made or substrate used up Dry Weight: organisms are filtered and then weighed to determine the starting material
41
Preserving Bacterial Cultures
* Refrigeration: for short term storage – what we use in laboratory * Deep freezing (-70C): long term (years) * Lyophilization (freeze drying): quick frozen cultures then placed under vacuum and sealed (decades)
42
Motility Medium
* Soft agar * Inoculate straight in with needle * Incubate at room temp If agar cloudy then microorganisms motile but if they are near injection point non-motile