Response of Staphylococcus Aureus to Acetylsalicylate Challenge While in the Presence Of

Response of Staphylococcus aureus to acetylsalicylate challenge while in the presence of Notatum Penicillium.(No Period)

Sarai Finks and Kazuhiro Sabet

Biology 3B, Department of Biology, Saddleback College, Mission Viejo, California

Bold -- Align flush with right margin Bacterial resistance to antimicrobial agents is emerging in a wide variety of pathways taken by pathogens. The ability of S. aureus to grow in an acetylsalicylate challenge was tested in the presence of N. penicillium. (After a 72-hour incubation period the mean diameter of the zone of inhibition around the sterilized chads under non-acetylsalicylic environment was 0.0 ± 0.0cm (± SEM), and the mean diameter around chads under an acetylsalicylic environment was 0.0 ± 0.0cm (± SEM).) Too wordy, run-on sentence The mean diameter around the 10mg of N. penicillium under a non-acetylsalicylic environment was 5.3 ± 0.4cm (± SEM), and the mean diameter around N. penicillium under an acetylsalicylic environment was 8.3 ± 0.6cm (± SEM). There is a significant difference between the four groups (p =???, ANOVA)(Running an ANOVA results in no significant difference, p=…). There was a significant difference in the mean diameter around the10mg of N. penicillium under an acetylsalicylic and non-acetylsalicylic environment was 5.5%. The mean diameter around the sterilized chads in S. aureus under an acetylsalicylic environment, and the mean diameter around the 10mg of N. penicillium in S. aureus under an acetylsalicylic environment was 14.2%. This shows a greater inhibition of S. aureus in the presence of a 30mM concentration with N. penicillium than with only the affect of N. penicillium on S. aureus alone.

Introduction

S. aureus has the ability to grow in high-salt comma and low moisture environments as well as resist multiple antibiotics in the presence of acetylsalicylate (aspirin) (Riordan, et al., 2006). S. aureus contains a strain that is methicillin-resistant, splice methicillin is in a group of antibiotics called beta-lactams (Lewis et al., 1990) and is a source for many enteric diseases. Investigators were interested in the antibacterial properties that many fungi contain and the properties that bacteria display in response to a threat. Penicillin for example is an endotoxin excreted from the (remove ‘the’) Penecillium fungi and has beneficial healing properties in that it was first synthesized for use as an antibiotic (Arriero 2002). The objective of this experiment was to observe the ability of Notatum Penecillium to withstand Staphylococcus aureus in the presence of acetylsalicylate (Riordan et al., 2006). (Did Riordan already conduct this experiment? Why quote them here?) Conflicting research exists as to whether acetylsalicylate inhibits or promotes bacterial growth in the presence of an antimicrobial (antimicrobial what?)(Arriero 2002, Lewis 2006). Acetylsalicylate is classified as a nonsteroidal antiinflammatory drug (Shiff et al.,1995). It is widley sp consumed for a variety of ailments ranging from pain to fever. It is also utilized as a means of preventative health measures against cancer and heart disease (Shiff et al., 1995). In this experiment acetylsalicylate was a successful growth inhibitor of S. aureus in the presence of N. Penecillium, (period) this observation may contribute to a better understanding of the properties and mechanisms of fungi in response to a bacterial threat. It is important to have a deeper understanding of how bacteria can adapt to overcome changes to their environment. Understanding how bacteria adapt and evolve to survive despite the best efforts of the present day antibacterial practices can possible aid in helping to prevent or treat infection (Tenover, 2001).

Materials and Methods

The bacteria S. aureus and fungus N. Penicillium (The S and N are italicized) were cultured over a 48-hour period. This was to ensure that the appropriate amounts of the organisms were available for plating on 27 plates. A nutrient agar was made by placing 27.6 g of nutrient starter in 1200 ml of water and carefully bringing the solution to a to a boil. Once the solution was autoclaved at 200º F for 30 minutes (The Autoclave utilizes a high pressure too. At what pressure?) (period) the solution was allowed to cool slightly and then poured over 27 sterile plates. The plates were allowed to cool and the cultures of S. areus sp(250 mL) and N. Penicillium (~10mg) were placed on the plates. The acetylsalicylate solution was made by dissolving 0.324 g of solid acetylsalicylic acid in 60.0 ml of DI water yielding a concentration of 30mM. Ten plates consisted of the S. aureus (250 mL), N. Penecillium, and acetylsalicylic acid (250 ml). Ten additional plates consisted of S. aureus and N. penecillium in the same concentrations as mentioned above(but no acetylsalicylate?). Seven plates where served (grammar)as the control. Sterilized chads were placed in DI water and also in the acetylsalicylate solution and placed on opposite ends of the same plate. This was done to observe the (effect)affect acetylsalicylate would have on bacterial growth. The plates were all placed in an incubator for 72-hours at 37º C. All temperatures and outside factors were held constant and data was then collected by measuring the halos or zone of inhibition (where bacteria did not grow) around N. penecillium and the chads.

Results:

Figure 1. The bar graph showing mean diameter of the zone of inhibition ± SEM around the sterilized chads and 10 mg of N. penicillium under an acetylsalicylic and non-acetylsalicylic environment. The figure represents the bacteria, S. aureus and the fungi, N. penicillium. (italicize figure caption)(figure should be after written results)

The measurements were taken after a 72-hour incubation period. The mean diameter of the zone of inhibition around the sterilized chads under non-acetylsalicylic environment was 0.0 ± 0.0cm (± SEM), and the mean diameter around chads under an acetylsalicylic environment was 0.0 ± 0.0cm (± SEM). The mean diameter around the 10mg of N. penicillium under a non-acetylsalicylic environment was 5.3 ± 0.4cm (± SEM), and the mean diameter around N. penicillium under an acetylsalicylic environment was 8.3 ± 0.6cm (± SEM). There is a significant difference between the four groups (p =, ANOVA).The Bonferroni correction showed no significant difference between the groups S. aureus in a non-acetylsalicylic environment and S. aureus in an acetylsalicylic environment was 0.0%. There was a significant difference in the mean diameter around the10mg of N. penicillium under an acetylsalicylic and non-acetylsalicylic environment was 5.5%. The mean diameter around the sterilized chads in S. aureus under an acetylsalicylic environment, (huh? Sentence fragment)and the mean diameter around the 10mg of N. penicillium in S. aureus under an acetylsalicylic environment was 14.2%. (what do all the % signs mean? aren’t these in cm? how do you get a ratio?)

Discussion:

Based on the results it is thought that there may be some reaction taking place between N. penicillium and the acetylsalicylate that caused a greater inhibition of bacterial growth of S. aureus since there is a difference, although the mechanism is not fully understood . This could be attributed to the specific proteins found with in the cell wall of N. Penecillium and related to the endotoxin that is released from the N. penecillium that gives it its antimicrobial traits. A reaction between the penicillin endotoxin and the acetylsalicylate could produce a greater toxic environment and thereby create a more unfavorable environment for S. aureus (italicize)to thrive than if just the penicillin were present. This is noted in the difference in the mean zone of inhibition for the bacterial growth and (remove ‘and’)which was 5.3 ± 0.4cm (± SEM) for the group with out the acetylsalicylate challenge and 8.3 ± 0.6cm (± SEM) for the group with the acetylsalicylate challenge. An ANOVA, p =, was then run for all the groups and showed a F-value equal to 100.5 showing a significant difference in the groups. The groups were then compared to each other by running a Bonferroni correction this revealed that there was a significant difference in the zones of inhibition between each group. The group containing S. aureus and N. penicillium and acetylsalicylate were significantly different than the group containing just the bacteria and the fungi and the group containing only the bacteria and the acetylsalicylate by 5.5% and 14.2% respectfully. Some sources for error may have arose and lead to error in measurements. These sources could be attributed to incorrect plating of the bacteria and the fungi and aspirin(acetylsalicylate). Cross contamination also played a minor role as some of the data had to be negated to over growth of fungus on the bacterial control. The control was very important since we were testing aspirins combined effect with penicillin to inhibit bacterial growth. The control was initially done but only qualitative observations could be made from it since sterilized chads had not been added. A second round of plates containing the bacteria and sterilized chads that had been treated with DI water and with a 30 mM solution of acetylsalicylate was used to quantitatively collect data. The same environment was created for the control and observations and data collected.

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Review Form

Department of Biological Sciences

Saddleback College, Mission Viejo, CA 92692

Author (s):______Sarai Finks and Kazuhiro Sabet______

Title:______Response of Staphylococcus aureus to acetylsalicylate challenge while in the presence of Notatum Penicillium______

Summary

Summarize the paper succinctly and dispassionately. Do not criticize here, just show that you understood the paper.

The study involved the effect acetylsalicylic acid on N. penicillium’s antimicrobial effects. Cultures of Staphylococcus aureus were plated with penicillium with and without acetylsalicylate. Results show a significant difference with the acetylsalicylate on the plates, widening the zone of inhibition by a significant amount. No significant difference was observed when S. aureus was plated with just the acetylsalicylate, and no penicillium, so the conclusion was made that the acetylsalicylate strengthens the antimicrobial effects of N. penicillium.

General Comments

Generally explain the paper’s strengths and weaknesses and whether they are serious, or important to our current state of knowledge.

The wording throughout is slightly confusing. Shortening the sentences in the introduction, results and discussion could help to clarify the study.

There are many misuses of commas, and many comma splices. Sentence structure should be fixed.

The figure’s x-axis should be clarified. Each Group should be clearly labeled.

The results for the zone of inhibition on the plate with S. aureus but no penicillium should not be described as 0.0 +/- 0.00 S.E.M. This should just be described as “no inhibition.” Discussing these numbers make one think its an important part of the study.

Discussion addresses the benefits of this test in the medical field. These findings are important and this is emphasized here.

Technical Criticism

Review technical issues, organization and clarity. Provide a table of typographical errors, grammatical errors, and minor textual problems. It's not the reviewer's job to copy Edit the paper, mark the manuscript.

This paper was a final version This paper was a rough draft XXX