MD4008: Introduction to Microbiology

Isolation of Aeromonas spp. from prepared foods

Aeromonas is an environmental bacterium which is detected in more than

500 cases of human gastrointestinal infection each year in England and Wales.
It can also infect mammals and birds (e.g. resulting in abortion in cattle;
septicaemia in young dogs). This study is focused on the potential presence of
Aeromonas spp. in ready-to-eat meals, and the isolation of bacteria that may
be Aeromonas spp. This handbook accompanies the study.
 You will use this handbook to complete a report on your findings. Your report
should consist of answers to the questions found within this handbook.

Direct support will be available on the module space, as well as model data
to inform your answers.

Part 1: Preparation and Plating of Samples

A packet of pre-packaged salad was obtained, and an attempt was made to

culture any bacteria present in the salad.

This involved plating out a portion of the salad, diluted in peptone saline
diluent (PSD), onto agar plates. These plates contained ampicillin, an antibiotic.
This provided any bacteria present with the opportunity to grow. After 48
hours individual colonies were selected from the agar, and these colonies were
spread on new agar, and again incubated for 48 hours. Once these new
colonies grew, they were studied as described in Part 2.

Part 1: Questions

1) Explain the role of Aeromonas spp. in human health, justifying the importance of this
study (510 words)(17 marks).

2) When plating out the salad samples, 2 agar plates were plated with pure PSD (containing
no salad sample). Explain why this was done (150 words)(5 marks).

3) Consider the role of ampicillin in the experiment: why is the antibiotic added to the agar
plates? (90 words)(3 marks).

Part 2: Gram Staining

One of the colonies produced in Part 1 was selected so the growing bacteria
could be further studied. The colony selected was studied using a Gram
staining protocol. The Gram stained colony was observed using light
microscopy, and appeared to be a pink colour.
 Part 2: Questions

4) Explain the purpose and scientific basis of the Gram stain, and interpret the results of the
Gram stain in the context of the study aim (450 words)(15 marks).

5) Identify any weaknesses of this study (150 words)(5 marks).

6) To confirm the identity of the isolated bacteria additional information would be required.
Suggest a future study that could confirm or refute the presence of Aeromonas spp. in the
packaged salad (150 words)(5 marks).