University of Phoenix Material

Appendix B – Identifying Bacteria

Bacteria identification is accomplished in a number of ways. Two common tools microbiologists use to identify unknown bacteria include dichotomous key and biochemical tests. The dichotomous key is useful when a microbiologist only needs to know which group an unknown microbe belongs to on a general level. When a microbiologist needs to identify a specific bacterium, biochemical tests are used.

PART ONE: GENERAL BACTERIA IDENTIFICATION

Review the dichotomous key in Figure A, the bacterial shapes in Figure B, and the Gram stain information below. You will use all three to determine to which major group unknown bacteria belong.

Figure B: The most common bacterial shapes. [Figure 4.1 in Microbiology text]

Shape Types: Comma, or Club-shaped, rods = Vibrio Rods = Coccobacillus, Bacillus Spherical = Coccus Spiral or Helical = Spirillium, Spirochete

Gram Stain Results:

Purple = Gram positive

Red = Gram negative

Neither purple nor red = No cell wall (neither Gram positive nor Gram negative)

Review Figure C on the following page. Use Figure C to identify the type of arrangement displayed by the unknown bacteria. Read through the Sample Identification on the following page for an example of how to identify bacterial groups and arrangements using Figures A, B, C, and the Gram stain results.

Figure C: Arrangement of Bacteria
Rods and Clubs
Single Strepto- Cording Snapping Palisades Chinese Letters
Cocci
Single Diplo- Tetrad Sarcinae Staphylo- Strepto-

Sample Identification

After performing a Gram stain, you observe the following under a microscope:

(Cells are stained red.)

· Begin with item 1 on the dichotomous key (Figure A). Because the organisms are red, they are not Gram-positive. According to the key, continue to item 3.

· Because the organism is red (not clear or another color), it is indeed Gram-negative according to item 3. The key directs you to item 5.

· Consider the shape of individual cells for item 5. According to Figure B, the shape type is closest to spherical. (Note: spherical is circular.)

· The bacteria belong to the Gram-negative cocci group.

· Now use Figure C to describe arrangement of bacteria. (The bacteria are not rod- or club-shaped, so focus on the cocci arrangements.) Identify the arrangement that is most prominent.

· The bacteria are arranged in a diplo- fashion.

· Enter the group and arrangement in the table.

Application

Use Figures A, B, C, and Gram stain results to identify group and arrangement of bacteria. Continue to Part Two after completing the table.

View from Microscope	Group of Bacteria	Arrangement
(stained red)	Gram-negative cocci	Diplo-
(stained purple)
(stained purple)
(stained red)
(stained red)
(stained purple)
(stained purple)

PART TWO: CAse study scenario: identify specific bacteria through biochemical testing

To gain an understanding of the processes involved with identifying bacteria through biochemical tests, access Chapter 6 of the text in WileyPlus located on the Week One course page. Once in WileyPlus (Chapter 6), select the “Bacterial Identification by API” link located under the heading, Take Another Look. Once selected, review the information and watch the Flash-animated movie (animation) located within this link. Then, review the following scenario and answer the questions that follow.

Case Study

A recent outbreak of food poisoning has occurred in a community. One possible source of contamination may be the produce that is grown and distributed locally at a farmer’s market. A test sample of some of the produce revealed evidence of bacterial contamination. The bacteria sample was tested in a microbiology lab and showed the results that follow. The laboratory also performed a Gram stain of the isolated bacteria and ran a number of biochemical tests to aid identification. The biochemical tests were assayed using the Analytical Profile Index (API) 20E system for identification of Enterobacteriaceae and other gram-negative bacteria.

Figure 1. Microbiology laboratory results – Unknown bacteria present on produce (API Results)

 

Figure 2. API Results of Unknown Bacteria in Text Format

Gram stain: Gram-negative

ONPG

ADH

LDC

ODC

CIT

H2S

URE

TDA

IND

VP

GEL

GLU

Bacteria: Unknown

+

–

+

+

–

–

–

–

+

–

–

+

MAN	INO	SOR	RHA	SAC	MEL	AMY	ARA
+	–	+	+	–	+	–	+

Based on Gram stain results and the knowledge that this bacterium caused food poisoning, the laboratory is able to narrow down the possibilities to three bacterial strains. Figure 3 shows the API results:

Figure 3. API Results of Known Bacterial Strains in Text Format

Gram stain: Gram-negative

ONPG

ADH

LDC

ODC

CIT

H2S

URE

TDA

IND

VP

GEL

GLU

Bacteria: Salmonella

–

–

+

–

–

–

–

–

–

–

–

+

MAN	INO	SOR	RHA	SAC	MEL	AMY	ARA
+	–	+	–	–	+	–	–

Gram stain: Gram-negative

ONPG

ADH

LDC

ODC

CIT

H2S

URE

TDA

IND

VP

GEL

GLU

Bacteria: Shigella

–

–

–

–

–

–

–

–

–

–

–

+

MAN	INO	SOR	RHA	SAC	MEL	AMY	ARA
–	–	–	–	–	–	–	–

Gram stain: Gram-negative

ONPG

ADH

LDC

ODC

CIT

H2S

URE

TDA

IND

VP

GEL

GLU

Bacteria: E. coli

+

–

+

+

–

–

–

–

+

–

–

+

MAN	INO	SOR	RHA	SAC	MEL	AMY	ARA
+	–	+	+	–	+	–	+

Application

Compare the API biochemical test results in Figure 2 (unknown bacteria from produce) with API test results in Figure 3 (known bacterial strains identified in the gray boxes on the left). The bacterial strain in Figure 3 that matches Figure 2 will identify the bacterial strain causing illness. Based upon your observations, which of the bacteria in Figure 3 is the most likely cause of the food poisoning outbreak?

©2008 University of Phoenix. All rights reserved.

Week One Worksheet: Natural Selection

Name: ___________________________________

Scenario One:

There are two types of beetles on the island of Arcandia – beetles that eat during the day (diurnal) and beetles that eat only at night (nocturnal). The birds on Arcandia seem to ONLY eat the diurnal beetles. During the day, the nocturnal beetles are inside the trunks of trees, escaping predation. In the springtime, each surviving female beetle lays about 500 eggs on the bottom side of the tree leaves, but only 100 of these eggs survive long enough to reproduce.

1. Identify the types of variation in this beetle population, and which beetles were selected for (natural selection is acting in their favor), and which were selected against (natural selection is acting against their favor) in the space below. Explain how you made this determination.

 

2. Determine which beetle type is most fit in its environment, and explain your choice using what you know about natural selection in the space below.

 

3. Predict how the beetle population will change over time, and explain how natural selection is working in this population of beetles in the space below.

 

 

Scenario Two:

In this scenario, there are two different colors of mice. Some mice are white in color, while others are dark brown. The main predator of the mice is the lynx. The lynx is able to catch about 80% more of the darker mice than the white mice because the white mice are better camouflaged. Each female mouse (white and brown) produces about 35 offspring per year.

1. Identify which mice were selected for, and which were selected against, and explain your choices in the space below.

 

2. Explain what would happen if the environment were to change, becoming warmer, and the snow melted in the space below. Describe how this would change which mice were selected for and against.

 

3. Predict how the mouse population will change over time, and explain how natural selection is acting on that population in the space below.

 

 

Page 2 of 2