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DID Case Study 2 Respiratory Infections

Intro
Sample D. Patient: Theresa Green
The patient was a 45 year old female. She underwent an unspecified transplant 2 weeks before her
admission on 12/03/13 with chest pains and shortness of breath. 3 days after admission a bronchial
washing was performed to provide a sample for testing for respiratory infectants.
From the bronchial wash several plate cultures were performed, as well as subcultures to ensure
isolation of a pure sample. Once the suspected organisms were isolated they were further tested to
identify the cause of the infection, and underwent antibiotic testing to provide a treatment
recommendation.
Method
1.5ml of the provided bronchial wash was centrifuged for 5 minutes. The resulting supernatant was
removed and the bacterial pellet was resuspended with a pipette. 3l of the suspension was fixed to
a microscope slide and underwent a gram staining. 3l of suspension was also used for each of 4
streak plates, 2 on nutrient agar and 2 on blood agar. These plates were incubated at 37C for 48
hours.
Using isolated colonies from the nutrient agar plates another gram stain was performed to confirm
the organisms were the same.
Next an oxidase test was performed to narrow down the search pattern for the API testing.
An API 20E was performed as per schedule to identify the organism.
Two spread subcultures were also performed on blood agar base plates, onto which M14 multidisks
were added to test for antibiotic sensitivity. These plates were incubated for 24 hours.
Results
The gram staining showed that the organism was a gram negative bacillius.
The colonies formed on nutrient agar were creamy white, smooth and slightly raised. This is
commonly idicative of E.coli.
Oxidase testing indicated that the organism was oxidase negative.
The API 20E result (7145553) identified the organism as either an Enterobacter spp. or E.coli.
The antibiotic sensitivity testing gave the following results:
Ampicillin Resistant
Cephalothin Resistant
Colistin sulphate Susceptible
Gentamicin Susceptible
Streptomycin Susceptible
Sulphatriad Resistant
Tetracycline Susceptible
Cotrimoxazole - Resistant
Due to the morphology of the colonies and the symptoms of the patient it was determined that the
organism was E.coli.
Given the results of the antibiotic sensitivity tests we would recommend the patient be treated with
either streptomycin or tetracycline as these were most effective in the lab.

Further Questions
1. As the patient was a recent transplant patient, they would have been immunosuppresed to
reduce the risk of transplant rejection. This will have left them vulnerable to infection from
any number of vectors. Gram negative organisms like E.coli are common causes of
nosicomial infection and it is possible that the patient picked it up while in the hospital. It
could also have been transmitted via infected food sources or through poor hygiene e.g hand
washing. The bacteria would then have been inhaled into the lungs causing upper respiritory
problems in the immunocompromised patient.
2.