The Effects of Different Strains of Pseudomonas aeruginosa on

Drosophila melanogastar

Need

http://knol.google.com/k/-/-/UtI7gr91/rIGkBA/Slide4.jpg

Knowledge BaseCharacteristics of

pseudomonas aeruginosa

• Tolerant to a wide variety of physical conditions

http://cmbi.bjmu.edu.cn/news/0310/21.htm

Knowledge Base

• Innate immunity is activated when microorganisms succeed in entering the body cavity of the fly.

• Invading microorganisms are killed by the combined action of cellular and humoral processes

• phagocytosed by specialized blood cells, surrounded by toxic melanin, or lysed by antibacterial peptides secreted into the hemolymph by fat body cells

Immune Response of Drosophila melanogastar

Knowledge Base

• enables bacteria to co-ordinate their behavior

http://www.quiestech.com/content/science/quorum-sensing_more.asp

Quorum Sensing

Knowledge Base

• Signal molecule used by pseudomonas aeruginosa

http://www.nottingham.ac.uk/quorum/what.htm

Quorum Sensing

Literature Review

• Stoltz, et al. (2008)– pricking the fly’s abdomen with a needle previously

dipped in a P. aeruginosa suspension – PON1 transgenic flies are protected from lethality due

to P. aeruginosa infection

Literature Review • D’Argenio, et al (2000)

– PAO1, the best-characterized strain of P. aeruginosa was used

– Adult female flies 2 to 4 days old were pricked in the dorsal thorax with a 25-gauge needle dipped directly into the PAO1 culture

Time course of infection of wild-type flies with PAO1 (  ), a pilD mutant (  ), and a chpA mutant (  ). Bacteria were grown as described for PAO1 and introduced into flies by pricking with a syringe needle. The chpA37 mutant was used for this and subsequent experiments. The values plotted are the averages of five replicate experiments, each with 10 flies, and the standard error of the mean is shown for each point.

Purpose

Hypothesis

The purpose of this experiment is to see if Drosophila melanogastar can be protected from the lethal strain of pseudomonas aeruginosa by being injected with the non-lethal strain first

Null hypothesis- The non-lethal strain of pseudomonas aeruginosa will have no effect on the protection of the flies from the lethal strain

Alternate hypothesis- The non-lethal strain will have an effect on the protection of the flies from the lethal strain

MethodologyDrosophila melanogastar

N=60

Flies put under anesthesia

Injected with non lethal strain

N=30

Injected with lethal strain

N=30

Injected with lethal strain

Amount of living flies are counted

Statistical analysis by ANOVA with p<.05

Protocols For Measurement

• The amount of dead or living flies will be counted manually

Do-Ability

• Flies are easily obtained, cheap, and easy to keep alive

• Bacteria can be kept alive using culturing techniques from articles that were read

Budget

Vendor Catalogue # Page Item Item # Quantity Unit $ Total $

Carolina Biological

FlyNap 173010 5 $12.50 $62.50

Frey Scientific

025 97 Drosophila Culture

563485016

1 $13.50 $13.50

Carolina Biological

Pseudomonas aeruginosaMicroKwik Culture®, Vial

155250A 1 $12.00 $12.00

$88.00

Bibliography