(Laboratory) Culture Media Preparation Flashcards
(106 cards)
1
Q
It is the media containing the required nutrients for bacterial growth.
A
Culture Media
2
Q
Serves as a soil where microbes can be planted and grown.
A
Culture Media
3
Q
Uses of Culture Media:
A
- Isolation and identification of microorganisms
- Performing anti-microbial sensitivity tests
4
Q
List down the Common Ingredients of Culture Media:
A
Peptone
Meat Extract
Yeast Extract
Mineral Salts
Carbohydrates
Agar
Water
5
Q
Extract which contains hydrolyzed plant and animal protein that serves as source of amino acid, peptides, and proteins for the bacteria.
A
Peptone
6
Q
A Common Ingredient of Culture Media that provides vitamins and mineral salts necessary for growth.
A
Meat Extract
7
Q
A Common Ingredient of Culture Media that serves as a growth stimulants.
A
Yeast Extract
8
Q
A Common Ingredient of Culture Media that provides inorganic compounds such as phosphates and sulfur necessary for bacterial metabolism.
A
Mineral Salts
9
Q
A Common Ingredient of Culture Media that serves as a source of energy necessary for the anabolic processes of the cell.
A
Cabohydrates
10
Q
_____ in TCBS Agar differentiates Vibrio spp.
A
Sucrose
11
Q
Sucrose in TCBS Agar differentiates _____
A
Vibrio spp.
12
Q
Sucrose in _____ differentiates Vibrio spp.
A
TCBS Agar
13
Q
_____ in MacConkey Agar differentiates Enterobacteria.
A
Lactose
14
Q
Lactose in _____ differentiates Enterobacteria.
A
MacConkey Agar
15
Q
Lactose in MacConkey Agar differentiates _____.
A
Enterobacteria
16
Q
A polymer that acts as the solidifying agent in culture media.
A
Agar
17
Q
An inert polysaccharide of seaweed.
A
Agar
18
Q
Property of Agar:
A
- It has high gelling strength
- High melting temperature (90-95 degree Celsius)
- Low gelling temperature
- It forms firm gel at 1.5% W/V concentration
- It forms semisolid gel at 0.4-0.5% W/V concentration
19
Q
Uses of Agar:
A
- Solidify culture media
- May provide calcium and organic ions to inoculated bacteria
20
Q
Preferred type of Water in Culture Media Preparation:
A
Deionized or Distilled Water
21
Q
Types of Culture Media based on Consistency:
A
- Liquid (Broth)
- Solid (Agar)
- Semisolid
22
Q
Types of Culture Media based on Nutritional Factors:
A
- Simple (Basal)
- Complex
- Synthetic or Defined
23
Q
List down the Special Media:
A
- Enriched
- Enrichment
- Selective
- Indicator or differential
- Transport
- Sugar media
24
Q
Selection of Culture Media will depend on:
A
- The major pathogens to be isolated, their growth requirements and the features by which they are recognized.
- Whether the specimens being cultured are from sterile sites or from sites having normal microbial flora.
- The cost, availability and stability of media.
- The training and experience of laboratory staff in preparing, using and controlling culture media.
25
A media that contains no solidifying agent or agar.
Liquid (Broth) Media
26
Growth in broth is signified by _____ or _____
Turbidity
Cloudiness
27
Uses of Liquid (Broth) Media:
1. As an enrichment media
2. As biochemical testing media
3. As blood culture media
28
This Type of Media is by adding a solidifying agent to the nutrients and water.
Solid (Agar) Media
29
Percentage of Agar needed on Solid Media:
2% to 3%
30
The most common solidifying agent is _____
Agarose
31
Growth in Solid (Agar) Media is indicated by the _____
Presence of Bacterial Colonies
32
Semisolid Culture Media contains how many percent of Agar?
0.5% to 1%
33
Uses of Semisolid Culture Media:
1. As an enrichment media
2. As motility media
34
A Type of Media that contain only basic nutrients required for the growth of ordinary organisms, and used as a general purpose media.
Basic/Simple/All Purpose Media
35
Uses of Basic/Simple/All Purpose Media:
1. To prepare enriched media
2. To maintain stock cultures of control bacterial strains
3. To subcuture pathogenic bacteria from selective/differential medium prior to performing biochemical or serological tests
36
A Type of Culture Media that contain some ingredients of unknown chemical composition.
Complex Media
37
One common ingredient of Complex Media is _____
Peptone
38
Types of Nutrient Broth:
1. Meat infusion broth
2. Meat extract broth
3. Digest broth
39
Nutrient Agar is prepared by adding agar at a concentration of 2% to the _____.
Nutrient Broth
40
Nutrient Agar is prepared by adding agar at a concentration of _____ to the nutrient broth.
2%
41
A Type of Culture Media wherein it is prepared exclusively from pure chemical substances and their exact composition is known.
Synthetic or Chemically Defined Media
42
A Type of Culture Media wherein it is prepared to meet the nutritional requirements of more bacteria by the addition of substances such as blood, serum or egg to a basal medium.
Enriched Media
43
Identify what type of Enriched Media based on the following characteristics given:
- Many bacteria are fastidious, requiring a medium that is even richer than nutrient agar commonly used in clinical laboratories.
- Used for isolation of Streptococci,
Pneumococci, Haemophilus
Blood Agar
44
Blood Agar is used for isolation of what group/s of species?
Streptococci
Pneumococci
Haemophilus
45
Identify what type of Enriched Media based on the following characteristics given:
- Used to culture even more fastidious bacteria.
- It is used for isolation of Neisseria (Meningococci and Gonococci) and Haemophilus.
Chocolate Agar
46
Chocolate Agar is used for isolation of what group/s of species?
Neisseria (Meningococci and Gonococci)
Haemophilus
47
Identify what type of Enriched Media based on the following characteristics given:
- Serum is added for enriching the medium.
- Used for the isolation of Corynebacterium diphtheriae.
Loeffler's Serum
48
Loeffler's Serum is used for isolation of what group/s of species?
Corynebacterium diphtheriae
49
A liquid media that is used to propagate the growth of certain groups of bacteria from a mixture of microorganism.
Enrichment Media
50
A Special Media in which the substance added to a liquid medium which inhibits the growth of unwanted bacteria and favors the growth of wanted bacteria. It is usually liquid and provides nutrients and environmental conditions that favor the growth of a particular microbe but not others.
Enrichment Media
51
An Enrichment Media that inhibits coliforms while allowing typhoid-paratyphoid bacilli to grow freely in fecal sample.
Tetrathionate Broth
52
An Enrichment Media that is used to isolate Salmonella spp. and Proteus spp. from stool samples.
Tetrathionate Broth
53
An Enrichment Media that is used for dysentery bacilli.
Selenite F (F for Feces) Broth
54
An Enrichment Media that is used for the isolation of Salmonella spp. from samples.
Selenite F (F for Feces) Broth
55
An Enrichment Media that is used for Vibrio cholerae from feces.
Alkaline Peptone Water
56
An Enrichment Media that is used to promote Vibrio spp.
Alkaline Peptone Water
57
Preferred pH Level of Alkaline Peptone Water:
8.5 pH
58
A Special Media in which the substance is added to a solid medium which inhibits the growth of unwanted bacteria but favors the growth of wanted bacteria. It is used to isolate particular bacteria from specimens where mixed bacterial flora is expected.
Selective Media
59
A Special Media that inhibits the growth of unwanted organisms while promoting the growth of the desired organism. This is achieved by the addition of inhibitors such as antibiotics, dyes, or chemicals.
Selective Media
60
A Selective Media that acts as a selective agent for dysentery bacilli (isolation of Shigellae).
Desoxycholate Citrate Agar (DCA)
61
A Selective Media that isolated Salmonella and Shigella from stools.
Desoxycholate Citrate Agar (DCA)
62
A Selective Media that supports the growth of Mycobacterium tuberculosis.
Lowenstein-Jensen Medium
63
A Selective Media that allows the growth of only Vibrio cholerae and inhibits the growth of intestinal organisms.
Thiosulfate Citrate Bile Salt (TCBS) Agar
64
A Type of Special Media/s that allows the visualization and differentiation of groups of bacteria usually based on the metabolic differences between each.
Indicator Media and Differential Media
65
Identify the Indicator Media based on the given:
- Incorporation of sulfite, S. typhi reduces sulfite to sulfide in the presence of glucose and the colonies of S. Typhi have a black metallic sheen.
Wilson and Blair Medium
66
Indicator on MacConkey Agar
Neutral Red Indicator
67
Identify the Indicator Media based on the given:
- Indicates lactose fermenting property, lactose fermenter (LF) produces pink colonies and non-lactose fermenter (NLF) produces colorless colonies due to a neutral red indicator.
MacConkey Agar
68
A Special Media that has substances incorporated in it, enabling it to bring out differing characteristics of bacteria and thus helping to distinguish between them.
Differential Media
69
_____ is both Differential and Selective; contains peptone, meat extract, NaCl, bile salt, lactose, and neutral red indicator.
MacConkey Agar
70
_____ is both Differential and Selective Media. It is Selective against gram-positve bacteria inhibiting their growth but it also differentiates members of the Enterobactericeae family as either non-lactose or lactose fermenter.
MacConkey Agar
71
Used to test fermentation.
Sugar Media
72
Used for the characterization and identification of bacteria, particularly important in the study of gram-negative bacilli.
Carbohydrate Fermentation
73
Sugars that are routinely employed for fermentation tests:
Glucose
Lactose
Sucrose
Mannitol
74
Components of Sugar Media:
1% of Sugar in Peptone Water along with an appropriate indicator
75
Indicator used in Sugar Media:
Anrade's Indicator - 0.005% Acid Fuchsin in NaOH
76
Color of the medium of a Sugar Media
Light Yellow
77
Sugar Media acid production is indicated by the development of _____
Pink Color
78
In Sugar Media, the gas accumulates in the inner _____
Durham's Tube
79
A small, inverted test tube placed inside a larger tube containing the media.
Durham's Tube
80
A Sugar Media that is used for organisms which are demanding in their growth requirements (fastidious organisms) like Streptococci, Pneumococci.
Hiss Serum Sugars
81
Culture Media used to determine sugar fermentation of fastidous organisms such as Neisseria and Streptococcus spp.
Hiss Serum Sugars
82
Media containing ingredients to prevent the overgrowth of commensals and ensure the survival of pathogenic bacteria when specimens can not be cultured soon after collection.
Transport Media
83
A Special Media that is used to preserve wanted organisms from the sample when there is delay between specimen collection and actual media inoculation.
Transport Media
84
Examples of Transport Media:
Amies Media
Stuart Media
Kelly-Blair Media
85
Special Media that is used to grow anaerobic organisms, and contain reducing substances.
Anaerobic Media
86
A Special Media that enhances the growth of anaerobic microorganisms
Anaerobic Media
87
A Media that acts as a reducing agent and creates an anaerobic environment deeper in the tube; contains reducing agents, such as sodium thioglycollate, glucose, vitamin C (ascorbic acid), cysteine and agar (concentration of 0.05%), with methylene blue.
Thioglycollate Broth
88
_____ or _____ acts as an oxidation reduction potential indicator, which should show that the medium is anerobic except in the surface layer in addition to a reducing agent and semisolid agar.
Methylene Blue
Resazurin
89
A Media that is used for the cultivation of aerobic, microaerophilic, and anaerobic microorganisms, especially Clostridium species; contains pieces of fat free minced cooked meat of ox heart and nutrient broth; supports the growth of both spore-forming and non-spore forming obligate anaerobes and also differentiate between putrefactive and saccharolytic species.
Cooked Meat Broth or RCM Broth (Robertson’s Cooked Meat Broth)
90
_____ of the meat indicates that the specie is saccharolytic while _____ of the meat indicates putrefaction.
Reddening
Blackening
91
List down the step-by-step procedure in the preparation of Culture Media:
1. Weighing and dissolving of culture media ingredients
2. Sterilization and sterility testing
- Addition of heat-sensitive ingredients
3. Dispensing of culture media
- pH testing of culture media
- Quality assurance of culture media
- Storage of culture media
92
A Sterilization Method that is used to sterilize most agar and fluid culture media.
Autoclaving
93
Preferred Time and Temperature for Autoclaving Sterilization Method:
121 degree Celsius, 15 psi for not less than 15 minutes
94
A Sterilization Method that is used to sterilize media containing ingredients that would be inactivated at temperature over 100 OC and re-melt previously bottled sterile agar media.
Steaming at 100 degree Celsius
95
A Sterilization Method that is used to sterilize additives that are heat-sensitive and can not be autoclaved.
Filtration
96
The simplest way to test for contamination is to incubate the prepared sample media at _____ for _____
35-37 degree Celsius
24 Hours
97
_____ in fluid media and _____ in solid media confirm contamination.
Turbidity
Microbial Growth
98
Using an aseptic technique, the ingredients should be added when the medium has cooled to _____, and should be distributed immediately unless further heating is required.
50 degree Celsius
99
The pH of most culture media is near neutral, and can be tested using _____ or _____
pH Papers
pH Meter
100
List down the ste-by-step method in Dispensing Agar Media in Petri Dishes:
1. Lay out the sterile petri dishes on a level surface.
2. Mix the medium gently by rotating the flask or bottle.
3. Flame sterilize the neck of the flask.
4. Pour 15 ml of the medium in each petri dish.
5. Stack the plates after the medium has gelled or cooled.
6. Store the plates in a refrigerator.
101
Plates of culture media, and additives like serum, blood and antimicrobials in solid form require storage at _____
2-8 degree Celsius
102
Antimicrobials in solution form should be stored at _____
-2 degree Celsius
103
All culture media and additives should be labeled with:
Name
Date of Preparation
104
The temperature selected for routine culturing is _____
35-37 degree Celsius
105
Anaerobic atmosphere is essential for the growth of strict anaerobes, and the techniques for obtaining anaerobic conditions are the following:
- Anaerobic jar with a gas generating kit
- Reducing agents in culture media
106
Anaerobic environment is achieved by using agents such as _____ that remove oxygen inside the chamber by combining it with hydrogen to form water
Palladium catalyst
