Frequency of Antibiotic Resistance among Bacteria Isolated From a High School Setting

Katie Amato

Columbia High School

Introduction

The number of intestinal infections is approaching 100 million cases per year in the United States.

Resistant bacterial strains often double hospital stay duration and mortality rates as compared to strains that can be treated.

Introduction

Antibiotic: a molecule used to kill Antibiotic: a molecule used to kill microorganisms and treat bacterial microorganisms and treat bacterial infectionsinfections

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Introduction

Resistance can be caused by DNA Resistance can be caused by DNA transfer via plasmids, or environmental transfer via plasmids, or environmental factors changing the DNA.factors changing the DNA.

A growing problem as the antibiotics we A growing problem as the antibiotics we have are becoming less and less effective. have are becoming less and less effective.

Introduction Antibiotics work by targeting different parts Antibiotics work by targeting different parts

of the bacteria cell and inhibiting proper of the bacteria cell and inhibiting proper function.function. If the antibiotic interferes with the synthesis of If the antibiotic interferes with the synthesis of

the cell wall, the bacteria will not reproduce, the cell wall, the bacteria will not reproduce, but will die.but will die.

For example, the interaction of penicillin with For example, the interaction of penicillin with penicillin-binding proteins.penicillin-binding proteins.

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Review of Literature

Mao, EF, Mao, EF, et al.et al. “Proliferation of mutators in a cell “Proliferation of mutators in a cell population.” population.” Journal of Bacteriology Journal of Bacteriology (Jan. 1997): (Jan. 1997): 417-422.417-422. Mutagens and mutationsMutagens and mutations

Giraud, Antione Giraud, Antione et al.et al. “Mutator bacteria as a risk “Mutator bacteria as a risk factor in treatment of infectious diseases.” factor in treatment of infectious diseases.” Antimicrobial Agents and Chemotherapy Antimicrobial Agents and Chemotherapy (Mar. (Mar. 2002): 863-865.2002): 863-865. Synergy treatmentSynergy treatment High concentrationHigh concentration

Review of Literature

Andersson, Dan. “Persistence of antibiotic resistant Andersson, Dan. “Persistence of antibiotic resistant bacteria.” bacteria.” Current Opinion in MicrobiologyCurrent Opinion in Microbiology 6 (2003): 6 (2003): 452-456.452-456. Fitness cost

Levy, Stuart B., and Marshall, Bonnie. “Antibacterial resistance worldwide: causes, challenges and responses.” Nature Medicine Supplement 10 (Dec. 2004): 122-129. Global problem Economic costs

Hypothesis

The majority of the bacteria found in this high school setting will not express antibiotic resistance.

Methods

Wet a sterile swab with sterile distilled water.

Thoroughly swab selected surface with the wetted swab and streak over a 5% Sheep’s blood agar plate.

Place plates in a 37° incubator overnight, or until colony growth appears.

Inoculated Control

Methods

After colony growth appears, isolate specific colonies off of the initial plate.

Use a sterile loop to pick up bacteria cells, and perform a three-point streak onto a different blood agar plate for each colony to be examined further.

Permanent stocks of cultures need to be made using a glycerol solution.

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Methods Gram staining will be used to determine

whether the bacterium is Gram positive or Gram negative.

Further testing

methods will

depend

on the Gram result. student.ccbcmd.edu

Mesacc.edu

Bd.com

Methods

Samples are to be tested for antibiotic sensitivity and/or resistance by exposure to different antibiotics (Kirby-Bauer Assay).

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Results—Plate 1

Sample 1-3 was found to be Gram+ cocci, catalase positive, and mannitol fermentation negative.

1-3

Results—Plates 2 and 3

Results—Plate 4 Sample 4-1 was found to be Gram- bacilli

and oxidase negative, but has not yet been identified using an Enterotube.

Results—Plate 6

6-1: Gram- bacilli, oxidase negative, yet to be Enterotube-d.

6-2: Gram- cocci, not yet identified6-1

6-2

Results—Kirby Bauer Assay

Discussion

Varying amounts and types of bacteria were found on all tested surfaces.

Samples need to be identified. 1-3, 6-1, and 6-2 are sensitive to the

tested antibiotics.

Conclusion

Of the bacteria that consistently grew in fresh culture, no resistance was found for the tested antibiotics.

The identity of the various bacterial samples remains unknown.

Future Research

Test more antibiotics Collect more samples Best results would be obtained from

freshly grown cultures.

Acknowledgements

Dr. Ann Zeeh Ms. Gleason and Ms. Strauss Ms. Masiello, Mr. Fudge, Mrs. Prout Science Research students My parents

References

Andersson, Dan. “Persistence of antibiotic resistant bacteria.” Current Opinion in Microbiology 6 (2003): 452-456.

Giraud, Antione et al. “Mutator bacteria as a risk factor in treatment of infectious diseases.” Antimicrobial Agents and Chemotherapy (Mar. 2002): 863-865.

Klugman, Keith, et al. “Bactericidal activity against cephalosporin-resistant Streptococcus pneumoniae in cerebrospinal fluid of children with acute bacterial meningitis.” Antimicrobial Agents and Chemotherapy 39 (6 Jul. 1995): 1988-1992.

Levy, Stuart B., and Marshall, Bonnie. “Antibacterial resistance worldwide: causes, challenges and responses.” Nature Medicine Supplement 10 (Dec. 2004): 122-129.

Mao, EF, et al. “Proliferation of mutators in a cell population.” Journal of Bacteriology (Jan. 1997): 417-422.

Paris, Maria, et al. “Management of meningitis caused by penicillin-resistant Streptococcus pneumoniae.” American Society for Microbiology 39.10 (1995): 2171-2175.

Sandora, Thomas, et al. “Reducing asenteeism from gastrointestinal and respiratory illness in elemntary school students: a randomized, controlled trial of an infection-control intervention.” Pediatrics 2008; 121; e1555. DOI: 10.1542/peds.2007-2597.

Questions?