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Antibiotic Resistance in Pathogenic Bacteria

Biol 115N- Lab Presentation
by

Ali Barstow

on 21 June 2013

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Transcript of Antibiotic Resistance in Pathogenic Bacteria

Colorimetric Assay
EXPERIMENT
Using Nitrocefin to test for TEM-1 activity in Gram-negative Bacteria
Results
Nitrocefin
Expected Outcome
Expected Outcome
antibiotic with a beta-lactam ring
cephalosporin:
changes from
yellow
to
red
in the presence of
TEM-1
Jennifer
PC
NC
Jennifer
PC
NC
Jennifer
PC
NC
If beta-lactamase
is present
If beta-lactamase
is NOT present
Antibiotic Resistance In Pathogenic Bacteria
Rod-Shaped Cells
with
Peritrichous Flagella
Production of
Hydrogen Sulfide
A. Barstow
B. Hamilton
M. Hosley
H. Strickland
Norfolk General Hospital - Patient Report
Name:
Jennifer
Gender:
F
Age:
28
Symptoms:
abdominal cramps
bloody diarrhea
fever
nausea
vomiting
Procedure:
Analyze Stool Sample
Gram-Negative
Unresponsive
to
Penicillin
Jennifer's Sample
Salmonella
Global concerns and challenges concerning antibiotic resistance:
higher death rates
prolonged illness
higher cost of medical care
harder to control spread of infection
jeopardizes health-care gains to society
Beta-Lactam Ring
is what makes
Penicillin effective
against Salmonella
Penicillin
Identifying Bacteria Strains
Why Size Matters
Gel
Electrophoresis
Each species of bacteria has DNA of a particular size
(measured in base pairs)
As it happens,
DNA has a negative (-) charge
If we put DNA
into a layer of gel, and place the gel into a magnetic field, the DNA will be pulled through the gel towards the positive (+) end of the field
The largest strands of DNA will move the slowest, and the smallest strands of DNA will move the fastest
EXPERIMENT
This joke is
in bad taste...
So, how do we
separate the
DNA to measure
it?
Get on over here, boy
Meh, maybe later...
Results of Gel Electrophoresis
After looking at the DNA samples under UV light, we found that the DNA from Jennifer's bacteria was the same size as DNA from a known strand of Salmonella.
Enterobacteriaceae
likes this.
These are the R plasmids
Negative control. No results shown.

Positive control
Jennifer's Sample
Ladder

~5,000 base pairs
TEM-1
The TEM-1 Gene codes for beta-lactamase enzyme
Fluoroquinolone
JENNIFER'S BACTERIA RENDERED THE PENICILLIN INEFFECTIVE
R-Plasmid
TEM-Gene resides on the
Resistant bacteria can "infect" other bacteria through
R-Plasmid Conjugation
Positive for beta-lactamase activity
Gel Electrophoresis
looking for the R-plasmid
Old Dominion University
Summer 2013
Biology 115N- Lab
A. Ackiss
Hypothesis:
Jennifer's symptoms were unresponsive to penicillin because she has a
resistant strain
of salmonella
Next Step:
Devise (2) experiments to determine the presence of TEM-1 beta-lactamase in Jennifer's sample
NOT a substrate for TEM-1!
Full transcript