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Frequency and Origin of an Gentamicin-Resistant E.

coli
Contamination in Lettuce at Ollie’s Organics
B. Greenspan, R. Prabhu, A. Pretory, H. Budowsky
Introduction
To understand a contamination of an unknown strain of antibiotic-resistant E. coli at Ollie’s Organics, the frequency of antibiotic
resistance and plasmid identity were analyzed. By comparing the populations of bacteria that grew on antibacterial plates to bacterial
growth on control plates, the frequency of antibiotic-resistance was determined as 0.321%. Gel electrophoresis was used for DNA
analysis and identified the plasmid as plasmid C and determined that it was isolated to the Ollie’s Organics farm.
Plasmid Causing Antibiotic Resistance Frequency of Antibiotic Resistance
Methods Methods
● Isolated antibiotic resistant bacteria from serial dilutions ● Serial dilution
● PCR ● Plate half on with antibiotics and half without antibiotics
● Gel Electrophoresis ● Count growth on each plate
Results Results
Paired
Our Data Average
DNA
Plasmid DNA Colony Positive control Plasmids Group
A B C
Ladder 1 2 3
Fig. 1: Gel electrophoresis 0.629% 0.012% 0.321%
of antibiotic resistant
bacteria at Ollie’s Organics.
Table 1: percentage of bacteria that grew on the antibiotic plate compared to the
The gel shows that our
non-antibiotic plate. This shows that roughly 0.3% of all of the E.coli from Ollie’s Organics
bacterial plasmid has antibiotic resistant bacteria.
corresponds with positive
control C, and it is the Discussion
smallest plasmid.
Concerning the first research question about which of the farms shows
evidence of E. coli contamination, the group results indicate that all of the
farms—Lucky’s Greens (L), Ella Farms (E), and Ollie’s Organics (O)—show
evidence of E. coli contamination due to at least some presence of
antibiotic-resistant bacteria on the antibiotic-containing culture plates.
Concerning the second research question about whether the bacterial
contamination at each farm is due to the same strain of bacteria or to different
strains, two different strains of antibiotic-resistant bacteria were found,
indicating a separation in contamination between farm O and farms L and E.
Recommendations Given that farms L and E both have bacteria with plasmid A, cross
● E and O are considered below low-level contamination contamination most likely must have occurred in distribution. Conversely, the
○ The precautionary principle dictates that they should still be bacteria from farm O had plasmid C, implying that farm O became
treated as low-level contaminated. contaminated independently during production. Concerning the third research
○ Romaine lettuce originating from farms E and O should only be question about what the frequency of the antibiotic resistant bacteria in the
used in products that require cooking or are pre-cooked, thus farm cultures were, farms E and O had very low levels of contamination (< 1%),
killing off any potential harmful bacteria (Manual, 2018). while farm L had a relatively higher—although still not very severe—level of
● Farm L is considered moderately contaminated; more intense steps contamination (~ 5%). According to the Recommendation Guidelines for
may need to be taken. Bacterial Contamination at Romaine Lettuce Farms, farms E and O technically
○ The romaine crop should be fully destroyed and the farm should be fall below Low-level contamination, while farm L falls on the lower edge of
placed under a 3 month probation program (Manual, 2018). moderate-level contamination (Manual, 2018).
○ WHO standard: it should be strongly considered whether farm L
should recall its romaine products (Angulo et al., 2008).
Advantages and Limitations of Study
● Farm O is believed to be independently contaminated and also has a ● Advantage: This study produced the first diagnosis of the severity of the
low frequency of antibiotic-resistant bacteria. contamination event, both through finding the relation between the strains
○ Mitigation can be contained to only crop production aspects. of bacteria in each farm, and the frequency of antibiotic-resistant bacteria in
● Farms L and E are potentially involved in cross-contamination. each farm culture.
○ Mitigation steps must be taken at their packaging and distribution ○ Using this study, the government can work to contain and mitigate the
levels. current outbreak
○ The precautionary principle dictates that crop production ● The frequency of contamination for each farm was surprisingly low for
mitigation also occur for both farms. farms E and O, yet high for farm L, suggesting possibly corrupted samples
○ The CDC recommendations that state laboratory capacity should despite group averaging.
be increased which will lead to more insight on the severity of ○ It is recommended that this study be repeated several times with new
contamination (CDC, 2018). samples in order to ascertain whether the results are truly accurate, as
well as researching the source of contamination for farms L and E.