The Effects of Different Strains of Pseudomonas aeruginosa on Drosophila melanogastar.
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Transcript of The Effects of Different Strains of Pseudomonas aeruginosa on Drosophila melanogastar.
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The Effects of Different Strains of Pseudomonas aeruginosa on
Drosophila melanogastar
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Need
http://knol.google.com/k/-/-/UtI7gr91/rIGkBA/Slide4.jpg
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Knowledge BaseCharacteristics of
pseudomonas aeruginosa
• Tolerant to a wide variety of physical conditions
http://cmbi.bjmu.edu.cn/news/0310/21.htm
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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
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Knowledge Base
• enables bacteria to co-ordinate their behavior
http://www.quiestech.com/content/science/quorum-sensing_more.asp
Quorum Sensing
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Knowledge Base
• Signal molecule used by pseudomonas aeruginosa
http://www.nottingham.ac.uk/quorum/what.htm
Quorum Sensing
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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
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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.
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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
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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
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Protocols For Measurement
• The amount of dead or living flies will be counted manually
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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
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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
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Bibliography