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micro para lab Flashcards

1
Q

Culture media

A

Food material or substances required for growing microorganisms in vitro
Artificial soil that contains nutrional and environmental requirements

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2
Q

Basic requirement of culture media

A

Water
Energy source
Carbon source
Nitrogen source
Salts
pH
Adequate oxidation
Growth factors

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3
Q

Chemotrophs

A

Organisms that obtain energy from chemical compounds

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4
Q

Chemoheterotrophs

A

Obtain energy from organic compounds
Most animals, fungi, and many bacteria

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5
Q

Chemoautotrophs

A

Obtain energy from inorganic compounds

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6
Q

Phototrophs

A

organisms that uses light for energy

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7
Q

photoautotrophs

A

convert carbon dioxide into organic compounds

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8
Q

photoheterotrophs

A

require organic compounds for carbon

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9
Q

Linotrophs

A

organisms that require specific nutrients

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10
Q

Organotrophs

A

utilize carbon sources like carbohydrates, lipids and proteins

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11
Q

Culture Media Based on Physical State

A
  • a- solid medium
  • b- semi-solid medium
  • c- liquid medium
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12
Q

Culture Media Based on the presence and absence of Oxygen

A
  • a- anaerobic media
  • b- aerobic media
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13
Q

Culture Media based on Nutritional Factors

A
  • a- simple medium
  • b- complex medium
  • c- synthetic medium
  • d- special medium
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14
Q

Solid medium

A

Agar is the most commonly used solidifying agent
-golden - yellow granular powder
- prepared from seaweeds
- not affected by the growth of the bacteria
- melts at 98degC & sets at 42degC

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15
Q

Semi-Solid Media

A

Lower agar concentration→0.2 - 0.5 %
Such media are soft and are useful in demonstrating bacterial motility and separating motile from non-motile strain
Useful in cultivation of microaerophilic bacteria

Hugh and Leifson’s oxidation fermentation medium
Stuart’s and Amies media
Mannitol motility media

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16
Q

Liquid Media

A
  • referred as broth
  • bacteria grow uniformly producing general turbidity
    - Nutrient Broth
    - Peptone Solution
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17
Q

Anaerobic Media

A

-cultivates anaerobic bacteria at low oxygen, reducing oxidation-reduction potential
-contains extra nutrients (Vit. K, Hemin, O2 that get reduced by physical or chemical processes

Robertson’s Cooked Meat Medium
Thioglycolate Broth Medium

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18
Q

Aerobic Media

A

-easy to cultivate microbes on solid media
-keeping cultures in incubator

liquid media, solid media
peptone water (1% peptone + 0.5% NaCl + 100mL water)
Nutrient Agar = Nutrient Broth + 2% agar

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19
Q

Candle Jar

A
  • a container into which a lit candle is introduced before sealing the container’s airtight lid
  • suitable for microaerophile (capnophiles) microorganisms that requires oxygen to survive, but requires environments containing lower levels of oxygen than are present in the atmosphere (20% concentration)
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20
Q

Simple Media

A
  • general purpose media that supports growth of non-fastidious microbes
  • primary used for isolation of microorganisms
    • Nutrient Broth, Nutrient Agar
      • NB consists of peptone, meat extract, NaCl
      • NB + 2% agar = Nutrient Agar
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21
Q

Complex Media

A

Blood Agar has ingredients that exact components are difficult to estimate

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22
Q

Synthetic Media or Defined Media

A

specially prepared media from pure chemical substances for research purposes and composition of every component is well known

peptone water→1% peptone + 0.5% NaCl in Water

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23
Q

Special Media

A
  • Enrich Media
  • Selective Media
  • Differential Media
  • Transport Media
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24
Q

Enrich Media (EM)

A
  • substances like blood, serum, egg are added to the simple medium
  • used to grow bacteria that are exacting in their nutritional needs
    • Fastidious Microorganisms (Neisseria gonorrhoeae)
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25
Q

Enrich Media
Blood Agar Plate (BAP)

A

contains mammalian blood (usually sheep or horse) typically at a concentration of 5-10%

used to isolate fastidious organisms and detect hemolysis

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26
Q

Enrich Media
Chocolate Agar

A

contain red blood cells that have been lysed by slowly heating to 80degC

used for growing fastidious bacteria such as Haemophilus influenzae

26
Q

Alpha Hemolysis

A
  • Partial lysis of red blood cells
  • Greenish discoloration around colonies
  • Commonly associated with Streptococcus pneumoniae
27
Q

Beta Hemolysis

A
  • Complete lysis of red blood cells
  • Appearance on blood agar: clear zone
  • Commonly associated with Streptococcus pyogenes
28
Q

Gamma Hemolysis

A

No lysis of red blood cells

29
Q

Selective Media

A

the inhibitory substance is added to a solid media to inhibit commensal or contaminating bacteria such as:
- antibiotics
- dyes
- chemicals
- alteration of pH

30
Q

Campylobacter Agar
selective media

A
  • used for isolation of Campylobacter jejuni for fecal or rectal swab
  • contain Bacteriological Charcoal, Cefoperazone and Amphotericin B
31
Q

Thayer Martin Medium
selective media

A
  • selective for Neisseria gonorrhoeae
  • usually contains the following combination of antibiotics:
    • Vancomycin→able to kill most Gram+ organisms
    • Colistin→added to kill most gram- organisms except Neisseria
    • Nystatin→can kill most fungi
    • Trimethoprim→inhibits gram- organisms, especially swarming Proteus
32
Q

Lowenstein - Jenson Medium
selective

A

-is solid medium for Mycobacterium tuberculosis
-contain:
- Penicillin
- Nalidixic Acid
- Malachite Green
in order to limit growth to Mycobacteria species only

33
Q

Eosin Methylene Blue (EMB)
selective media

A
  • selective for gram- bacteria
  • the dye methylene blue in the medium inhibit the growth of gram+ bacteria
34
Q

Differential Media

A

designed in such a way that different bacteria can be recognized on the basis of their colony color

dyes and metabolic substrates are incorporated so that those bacteria that utilize them appear as differently colored colonies

35
Q

MacConkey Agar

A
  • distinguish between:
    • Lactose Fermenters → Pink Colonies (Escherichia coli, Enterobacter aerogenes)
    • Non-Lactose Fermenters → colorless colonies
      (Proteus vulgaris, Salmonella typhimurium, Staphylococcus auerus)
36
Q

Cystine-Lactose-Electrolyte-Deficient Agar
CLED

A
  • Used for urinary tract infections , urine specimen
  • Inhibits swarming of Proteus species
  • Supports growth of Gram-negative bacteria
36
Q

Mannitol Salt Agar

A
  • mannitol fermenters includes:
    • Staphylococcus aureus
  • non-mannitol fermenters:
  • positive growth but mannitol non-fermenters:
    • Micrococcus luteus
  • negative growth includes:
    • Escherichia coli
    • Pseudomonas aeruginosa
37
Q

Thiosulfate-citrate-bile salts-sucrose agar
TCBS

A
  • Highly selective for the isolation of V. cholerae and V. parahaemolyticus
  • Inhibits Gram-positive bacteria
  • Differentiates sucrose fermenters
  • Color change indicates fermentation
37
Q

Xylose Lysine Deoxycholate Agar
XLD

A

Selective for the isolation of Salmonella and Shigella spp from clinical specimens and from samples

  • Black colonies- Salmonella sp.
  • Yellow colonies- Escherichia coli
38
Q

Transport Media

A

used for transporting the samples

delicate organisms may not survive the time taken for transporting the specimen without a transport media

  • Stuart’s medium
  • Buffered Glycerol Saline
38
Q

Plant Tissue Culture

A

Murashige and Skoog (also called MS media, MSO, or MS0) and Gamborg’s B5 medium

39
Q

information written on the bottom of the plate before inoculation:

A
  • date of inoculation
  • temperature of incubation
  • duration of incubation
  • microorganism inoculated
40
Q

streak plate

A
  • common method for isolation of a pure culture from a mixture
  • the Inoculum is the streaked over the agar surface to isolate colonies on at least a portion of the plate
  • ideal results require that only a small amount of inoculum
41
Q

4 types of streaks

A

1- quadrant
2- T- streak
3- continuous
4- radiant

42
Q

Quadrant streaking

A
  • area 1- heavy confluent growth
  • area 2- less dense
  • area 3- weak growth
  • area 4- isolated colonies
43
Q

T- streaking

A
  • 1st sector- heavy confluent growth
  • 2nd- less heavy growth, no isolated colonies
  • 3rd- isolated colonies
44
Q

Continuous streak

A

inoculum is evenly distributed

45
Q

Radiant streak

A

suitable to propagate pure culture

46
Q

Zigzag Streaking

A

commonly done to propagate the pure culture and culture them in large quantities

47
Q

Serial Dilution

A
  • is the stepwise dilution of a substance in solution
  • usually the dilution factor at each step is constant, resulting in a geometric progression of the concentration in a logarithmic fashion
48
Q

Spread Plate Method

A
  • is a technique to plate a liquid sample containing bacteria so that the bacteria are easy to count and isolate
  • used for viable count
49
Q

Pour Plate Technique

A
  • can be used to determine the number of microbes/mL in a specimen
  • it has the advantage of not requiring previously prepared plates and is often used to assay bacterial contamination of foodstuffs
  • used for viable count
50
Q

Stab Culture

A
  • by puncturing a suitable medium with a long straight charged wire
  • for gelatin liquification, stock cultures and motility
  • deep agar
51
Q

Lawn Culture

A

provides a uniform surface growth of the bacterium
uses:
- for bacteriophage typing
- antibiotic sensitivity
- in the preparation of bacterial antigens and vaccines

52
Q

Semi solid mediums

A

Hugh & Leifson’s oxidation fermentation medium
Stuart’s & Amies media
Mannitol motility media

53
Q

Mannitol fermenter

A

Staphylococcus aureus

54
Q

Non mannitol fermenters

A

+ growth
Micrococcus luteus

  • growth
    E. coli
    Pseudomonas aeruginosa
55
Q

Anaerobic mediums

A

Robertson’s cooked meat
Thioglycolate broth

56
Q

Vancomycin

A

Able to kill most gram + organisms

57
Q

Colistin

A

Added to kill most gram- organisms except Neisseria

58
Q

Nystatin

A

Can kill most fingi

59
Q

Trimethoprim

A

Inhibits gram- organisms specially swarming proteus

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