Need help with questions 1) Evaluate the data for each organism. Is each organism equally impacted by the chemotherapeutic agents listed? Use your data to support your answer. 2) When you compare the data for the Gram-positive cell to the data for the Gram-negative cell, can you say there is similar impact? Use your data to support your answer. 3) Which antibiotic(s) is/are more effective against the Gram-positive cell type? Gram-negative cell type? Use your data to support your answers. 4) Which antibiotic(s) is/are least effective against the Gram-positive cell type? Gram-negative cell type? Use your data to support your answers.

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Need help with questions 1) Evaluate the data for each organism. Is each organism equally impacted by the chemotherapeutic agents listed? Use your data to support your answer. 2) When you compare the data for the Gram-positive cell to the data for the Gram-negative cell, can you say there is similar impact? Use your data to support your answer. 3) Which antibiotic(s) is/are more effective against the Gram-positive cell type? Gram-negative cell type? Use your data to support your answers. 4) Which antibiotic(s) is/are least effective against the Gram-positive cell type? Gram-negative cell type? Use your data to support your answers.
Antibiotic Sensitivity with the Gram Stain as Background Information
The Gram Stain is the most important stain used in microbiology. Knowing whether a bacterial
cell is Gram-positive or Gram-negative determines the type of antibiotic used to treat an
infection. There are antibiotics that target Gram-positive cells, there are antibiotics that target
Gram-negative cells, and then there are those that target both Gram-positive and Gram-negative
cells (e.g. broad-spectrum antibiotics).
Danish bacteriologist Hans Christian Gram developed the Gram Stain in 1884. The Gram Stain
is a staining technique that separates bacteria into the two groups mentioned above. It is based on
the ability of bacteria to retain the purple color of crystal violet during decolorization with
alcohol. Gram-positive bacteria remain purple after decolorization and Gram-negative bacteria
are decolorized as they lose the purple color. To visualize the Gram-negative cells, the cells are
counterstained with safranin. After staining with safranin, the cells look pink. See below:
Rod
Gram Stain
(gram-negative)
Celer ef
Gram-Posve Cell Graegative Cel
Celer o
Primary Stain
Crystal Vielet
Purple
Purple
Cocei
Mordant
Purple
Purple
Igram pesitive
ladine
Decalarizing Agent
ANhelAcetone
Purple
Caloress
Counterstain
Purple
RedPrk
Satranin
(b)
Microbiology, An Introduction, 11 cd., Tortora et al., Pearson, 2013
The two cell types differ structurally in the following way:
Figure 4.130eBecterial cell walls.
Peptidogycan
Well teichoic acid
Cell wall
LUpoteichoic
acid
Plasma
membrane
Protein
O polysaccharide
Core pelyaccharide
(b) Gram-positive cell wll
Core polysaccharide
Lipopatycharide o polyaccharide
Lipid A
Parta of the LPS
Porin protein
Lipoprotein
Cell wa Outer membrane
LPeptideglycan
Plasma
mebrane
Phospholipid
Periplasm
Protein
(e) Gramegtive cell wall
arnt
The video link below is a short presentation on the cellular differences between the two cell
types. Please access and watch for clarification on the figure above.
https://www.bing.com/videos/search?q=gram+positive+vs+gram+negative+bacteria&ru=%2fvid
cos%2fsearch%3fq%3dgram%2bpositive%2bvs%2bgram%2bnegative%2bbacteria%26FORM%
%203dVDVVXX&view detail&mid-E52E108AA9D86F20114CE52E108AA9D86F20114C&
&FORM=VDRVRV
The exercise will focus on the analysis of antibiotic impact on a Gram-positive cell type,
Staphylococcus aureus, and Gram-negative cell type, Escherichia coli. These are not the only
species that exhibit similar behavior in the presence of antibiotics. Other examples of bacteria are
shown below.
Gram positive
bacteria
Gram negative
hacteria
Transcribed Image Text:Antibiotic Sensitivity with the Gram Stain as Background Information The Gram Stain is the most important stain used in microbiology. Knowing whether a bacterial cell is Gram-positive or Gram-negative determines the type of antibiotic used to treat an infection. There are antibiotics that target Gram-positive cells, there are antibiotics that target Gram-negative cells, and then there are those that target both Gram-positive and Gram-negative cells (e.g. broad-spectrum antibiotics). Danish bacteriologist Hans Christian Gram developed the Gram Stain in 1884. The Gram Stain is a staining technique that separates bacteria into the two groups mentioned above. It is based on the ability of bacteria to retain the purple color of crystal violet during decolorization with alcohol. Gram-positive bacteria remain purple after decolorization and Gram-negative bacteria are decolorized as they lose the purple color. To visualize the Gram-negative cells, the cells are counterstained with safranin. After staining with safranin, the cells look pink. See below: Rod Gram Stain (gram-negative) Celer ef Gram-Posve Cell Graegative Cel Celer o Primary Stain Crystal Vielet Purple Purple Cocei Mordant Purple Purple Igram pesitive ladine Decalarizing Agent ANhelAcetone Purple Caloress Counterstain Purple RedPrk Satranin (b) Microbiology, An Introduction, 11 cd., Tortora et al., Pearson, 2013 The two cell types differ structurally in the following way: Figure 4.130eBecterial cell walls. Peptidogycan Well teichoic acid Cell wall LUpoteichoic acid Plasma membrane Protein O polysaccharide Core pelyaccharide (b) Gram-positive cell wll Core polysaccharide Lipopatycharide o polyaccharide Lipid A Parta of the LPS Porin protein Lipoprotein Cell wa Outer membrane LPeptideglycan Plasma mebrane Phospholipid Periplasm Protein (e) Gramegtive cell wall arnt The video link below is a short presentation on the cellular differences between the two cell types. Please access and watch for clarification on the figure above. https://www.bing.com/videos/search?q=gram+positive+vs+gram+negative+bacteria&ru=%2fvid cos%2fsearch%3fq%3dgram%2bpositive%2bvs%2bgram%2bnegative%2bbacteria%26FORM% %203dVDVVXX&view detail&mid-E52E108AA9D86F20114CE52E108AA9D86F20114C& &FORM=VDRVRV The exercise will focus on the analysis of antibiotic impact on a Gram-positive cell type, Staphylococcus aureus, and Gram-negative cell type, Escherichia coli. These are not the only species that exhibit similar behavior in the presence of antibiotics. Other examples of bacteria are shown below. Gram positive bacteria Gram negative hacteria
shown below.
Gram positive
bacteria
Gram negative
hacteria
dreamstime.com
The representative data set you will work with would have been acquired after inoculating two
plates of medium (one for each organism) and placing disks permeated with varied antibiotics.
The procedure for generation is as follows (source: www.slideshare.net):
How to perform Kirby- Bauer
testing
• The basics are easy: The bacterium is
swabbed on the agar and the antibiotic
discs are placed on top. The antibiotic
diffuses from the disc into the agar in
decreasing amounts the further it is away
from the disc. If the organism is killed or
inhibited by the concentration of the
antibiotic, there will be NO growth in the
immediate area around the disc: This is
called the zone of inhibition
Each plate what have looked like the one below (source: www.researchgate.net). The Zone of
Inhibition is what you would have measured in mm. The mechanism for the Zone of Inhibition is
either death or growth inhibition. This type of testing does not allow the distinction between a
| 2
bactericidal (killing) or bacteriostatic (not allowing population expansion) mechanism. What is
measured is the clear area from the edge of the disk to the start of the growth. The greater the
clearing, the greater the impact. The lack of clearing means no impact.
Data:
Staphylococcus aureus (G+)
20 mm
Escherichia coli (G-)
22 mm
Antibiotic Used
Chloramphenicol
Erythromycin
Penicillin
Streptomycin
Sulfamethoxazole
Tetracycline
Triple Sulfa
Vancomycin
Protein Synthesis Inhibitor; Cell Wall Synthesis Inhibitor (Peptidoglycan Impact);
Metabolic Inhibitor
12 mm
5 mm
1 mm
8 mm
2 mm
18 mm
6 mm
3 mm
19 mm
O mm
17 mm
20 mm
O mm
O mm
Questions:
1) Evaluate the data for each organism. Is each organism equally impacted by the
ch
Transcribed Image Text:shown below. Gram positive bacteria Gram negative hacteria dreamstime.com The representative data set you will work with would have been acquired after inoculating two plates of medium (one for each organism) and placing disks permeated with varied antibiotics. The procedure for generation is as follows (source: www.slideshare.net): How to perform Kirby- Bauer testing • The basics are easy: The bacterium is swabbed on the agar and the antibiotic discs are placed on top. The antibiotic diffuses from the disc into the agar in decreasing amounts the further it is away from the disc. If the organism is killed or inhibited by the concentration of the antibiotic, there will be NO growth in the immediate area around the disc: This is called the zone of inhibition Each plate what have looked like the one below (source: www.researchgate.net). The Zone of Inhibition is what you would have measured in mm. The mechanism for the Zone of Inhibition is either death or growth inhibition. This type of testing does not allow the distinction between a | 2 bactericidal (killing) or bacteriostatic (not allowing population expansion) mechanism. What is measured is the clear area from the edge of the disk to the start of the growth. The greater the clearing, the greater the impact. The lack of clearing means no impact. Data: Staphylococcus aureus (G+) 20 mm Escherichia coli (G-) 22 mm Antibiotic Used Chloramphenicol Erythromycin Penicillin Streptomycin Sulfamethoxazole Tetracycline Triple Sulfa Vancomycin Protein Synthesis Inhibitor; Cell Wall Synthesis Inhibitor (Peptidoglycan Impact); Metabolic Inhibitor 12 mm 5 mm 1 mm 8 mm 2 mm 18 mm 6 mm 3 mm 19 mm O mm 17 mm 20 mm O mm O mm Questions: 1) Evaluate the data for each organism. Is each organism equally impacted by the ch
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