BIOL 1020 Unit 1 Module 3 Lab Assessment

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Using the VUMIE Lab Manual, the VUMIE website, your notes, and in some
cases outside research, answer the following questions. Your answers must be in
the form of complete and grammatically correct sentences. Highlight your
responses or use a different font color (to distinguish them from the questions
themselves). These questions are ordered as you proceed through the VUMIE
lab exercise sections. 10 points
Growth Media and Patterns of Growth
1) Click on the M icon to find reference material on the various media
included in VirtualUnknown Microbiology, and create and complete a
table that looks like the table in Question 2 on page 25 of your lab manual.
Complex or Synthetic? Selective, Differential, Both, Neither?

Malonate broth

Synthetic

Both

Tryptone broth

Complex

Differential

KCN broth

Synthetic

Selective

OF glucose broth

Complex

Differential

Mannitol salt agar

Synthetic

Both

Simmons citrate agar Synthetic

Both

2) The bacteria for this unknown are Gram negative rods. Compare the
media in the table above with the dropdown list of media available for

BIOL 1020 Unit 1 Module 3 Lab Assessment

conducting tests with Gram negative rods. Which of the media listed in the
table is/are used for isolating or identifying Gram positive cocci?
The media listed in the table that is used for isolating or identifying Gram positive
cocci is Mannitol Salt Agar.
3) How does the selectivity of mannitol salt agar compare/contrast to the
selectivity of malonate broth with regard to (1) Gram positive vs. Gram
negative, and (2) among various Gram negatives?
Malonate broth is used to determine whether the microbe can use the compound
malonate for carbon and energy. Mannitol salt agar is selective in allowing
bacteria capable of growth in 7.5% NaCl while preventing the growth of salt
sensitive microbes. The selectivity of Mannitol salt agar identifies Gram-positive
cocci. The Mannitol salt agar test is used to identify Gram-positive bacteria. The
7.5% NaCl will inhibit the growth of most Gram-positive and Gram-negative
bacteria.
4) Click the M button to get to the Media resources and use the info
provided to create and complete a table that looks like the table in
question 5 on page 26 of your lab manual.
Carbon Source

Nitrogen Source

Phenol red adonitol broth

Adonitol

Proteose peptone

Malonate broth

Malonate

Ammonium sulfate

Christiansens urea broth

Urea

Urea, Ammonia

KCN broth

Potassium cyanide

Potassium cyanide

5) Complete the OF glucose tests for your unknown bacterium by

inoculating, incubating, and interpreting the results using OF glucose broth
and OF glucose broth with oil overlay.
a. Based on these results, highlight or underline below the type of
metabolism that best fits your unknown microbe:

BIOL 1020 Unit 1 Module 3 Lab Assessment

Oxidation of glucose
Fermentation of glucose
Both oxidation and fermentation
Neither
6) Another factor influencing growth is incubation temperature. Extremes of
heat and cold can slow or halt growth altogether. Provide examples from
your own kitchen of the use of extremes in heat and cold to slow or
prevent microbial contamination of foods.
Examples from my own kitchen of the use of extremes in heat to slow or
prevent microbial contamination of foods include boiling water and cooking
food on the stovetop to kill contaminated microbes. Examples of extreme
cold include freezing foods to slow the contamination.
7) The results of growth can be followed using a spectrophotometer to detect
how much light can pass through a broth culturethe more cells, the less
light passes through. Resulting growth curves are described in your text.
In a growth curve, the number of cells grows logarithmically
(exponentially) until a maximum number is achieved the maximum
number that can be sustained by the medium (called maximum stationary
phase). Growth conditions that are not optimal result in slower rates of
growth. Using this information, predict the growth rates expected results
after 18 hours for six sterile tubes of an identical medium inoculated
simultaneously with an enteric microbe and incubated at 15C, 25C,
35C, 45C, and 55C. Use the symbols -, [-], =, [+], and + (these are in
order of increasing growth) to indicate growth relative to the maximum
growth rate at 37C (where, if graphed, the slope of the exponential
portion of the growth curve is greatest) and fill in the table below. The
symbols are used to indicate growth amounts from lowest to highest:

- < [-] < = < [+] < +

So, [+] is used to indicate somewhat less growth than a +. Lack of growth
is indicated by a -, while some growth (not a great deal) is indicated with
the [-]. All of this is relative to what should be the optimal growth
temperature for an enteric bacterium
Refer to the graph on page 27 of your lab manual to help predict these
results.
Temperature (C)
15C

Expected Growth Rate

After 18 Hours (-, [-],
=, [+], or +)
[-]

BIOL 1020 Unit 1 Module 3 Lab Assessment

25C

[+]

35C

45C

[+]

55C

[-]