Dizon, Diana A. BSN 3-3: Macconkey'S Agar (Mac) Is A Medium That Tends To Select For The Growth of

DIZON, Diana A.
BSN 3-3
QUESTIONS TO ANSWER
Briefly answer the following questions and upload your answers in our class Gdrive.
1. Why are complex media preferable to chemically defined media for routine
cultivation of microorganisms?
Complex media usually contain complex materials of biological origin such as

blood or milk or yeast extract or beef extract, the exact chemical composition of
which is obviously undetermined; they are more preferable for routine cultivation of
microorganisms than chemically defined media because it provides highly enriched
sources such as plant or animal extracts, vitamins, minerals, protein, sugar, and or
blood to the organism, supplying them specific nutritional substances needed for
their growth. Chemically defined media on the other hand does not supply such a
vast abundance of nutritional substances
2. Give two examples of differential media and indicate the purpose of each medium.
 MacConkey’s agar (MAC) is a medium that tends to select for the growth of
Gram-negative organisms while inhibiting the growth of Gram-positives. At
the same time, it is a differential medium that allows for the differentiation of
organisms that can ferment lactose from those that cannot. MacConkey’s
Agar is recommended for use in the detection and isolation of all types of
dysentery, typhoid, and paratyphoid bacteria (Salmonella). It promotes the
development of these organisms and at the same time differentiates them
from lactose-fermenting Gram-negative bacilli such as Escherichia coli.
Isolated colonies of lactose fermenting bacteria (coliforms) are opaque pink
in color, while Salmonella colonies are uncolored and translucent.
Differentiation is most clear with well-isolated colonies. The pH indicator
present is neutral red, and accumulates within and sometimes around
colonies able to ferment lactose.
 Mannitol Salt Agar (MSA) can be used to presumptively isolate and identify
Staphylococci from human samples. Refer to the compositions of MSA and
MacConkey agar below. MSA contains 75 g/L NaCl (7.5%) compared to
the 5 g/L found in TSA and other all-purpose media. MSA favors the growth
of salt tolerant microbes, namely Staphylococci, because other bacteria
from a human sample, are inhibited by the high NaCl component. In
addition, to distinguish pathogenic Staphylococci, namely Saureus from
other common Staphylococci, the substrate mannitol (a sugar) and the pH
indicator phenol red are added. If the organism ferments mannitol, acids
will be produced as byproducts. These acids will lower the pH changing the
indicator from pink to yellow. S. aureus can ferment mannitol, while other
common Staphylococci found in humans cannot.
3. You are asked by your laboratory instructor to prepare nutrient agar for your group.
According to the manufacturer’s instruction, 28 grams of nutrient agar powder
should be suspended in 1 liter of distilled water. How many grams of nutrient agar
powder is needed to prepare 10 big agar slants and 15 agar plates? Show your
solution.
Formula:
Wt. Of media in “Directions(g) = Wt. Of powder should we
weight
V of D.W in “Directions” (ml) V of media (D.W) we need (ml)
10 big agar slants x 8 = 80ml

15 agar plates x 20 ml = 300ml
= 380 ml
28 grams = x (28g) (380ml) = (1,000ml) x

1,000 ml 380 ml 1,000ml 1,000ml
Wt. Of powder should we weight = 10.64 grams
4. Tryptic Soy Agar (TSA) is a general growth medium used for the isolation and
cultivation of microorganisms. You need to prepare 20 agar plates, 5 big agar
slants and 15 small agar slants. Since dehydrated TSA is not available in the lab,
you searched for its components. The table below shows the composition of TSA
per liter of solution.
INGREDIENTS GRAMS/LITER
Casein peptone 17.0

Soya peptone 3.0
Sodium chloride 5.0
Dipotassium hydrogen phosphate 2.5
Glucose 2.5
Agar 15
How much of each component is needed to prepare the medium for the whole class?
Show your calculation.
Formula: Wt. Of media in “Directions(g) = Wt. Of powder should we

weight V of D.W in “Directions” (ml) V of media (D.W) we
need (ml)
17grams/liter 0.017 g/ml
3.0grams/liter 0.003g/ml
20 agar plates x 20 = 400ml

5 big agar slants x 8 = 40ml
15 small agar slants x 3 = 45ml
x = grams = 485ml
0.017g =x (0.017g) (485ml) = 8.245 g of Casein peptone

ml 485ml ml
0.003g =x (0.003g) (485ml) = 1.455 g of Soya peptone

ml 485ml ml
0.005g =x (0.005g) (485ml) = 2.425 g of Sodium chloride
ml 485ml ml
0.0025g = x (0.0025g) (485ml) = 1.2125 g of Dipotassium hydrogen

phosphate
ml 485ml ml
0.0025g =x (0.0025g) (485ml) = 1.2125 g of Glucose

ml 485ml ml
15g =x (15g) (485ml) = 7.275 of Agar

1000ml 485ml 1000ml
5. Provide one advantage and disadvantage for each type of sterilization methods.
REFERENCES
http://textbookofbacteriology.net/nutgro_3.html
https://biosci.sierracollege.edu/materials/4/laboratory_syllabus/use_of_selective_and_
differential_media.pdf
https://mospace.umsystem.edu/xmlui/bitstream/handle/10355/69341/3.2-
SDM1%20Selective%20and%20Differential%20Media%201.pdf?
sequence=13&isAllowed=y
https://www.researchgate.net/figure/Advantages-and-disadvantages-of-sterilization-
techniques_tbl4_303325664

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DIZON, Diana A.

Complex media usually contain complex materials of biological origin such as

10 big agar slants x 8 = 80ml

28 grams = x (28g) (380ml) = (1,000ml) x

Casein peptone 17.0

Formula: Wt. Of media in “Directions(g) = Wt. Of powder should we

17grams/liter 0.017 g/ml

20 agar plates x 20 = 400ml

0.017g =x (0.017g) (485ml) = 8.245 g of Casein peptone

0.003g =x (0.003g) (485ml) = 1.455 g of Soya peptone

0.0025g = x (0.0025g) (485ml) = 1.2125 g of Dipotassium hydrogen

0.0025g =x (0.0025g) (485ml) = 1.2125 g of Glucose

15g =x (15g) (485ml) = 7.275 of Agar

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