11 thoughts on “P17 – Dittmer

    1. It means that we took phage from one plaque and made a lysate with it (which was a liquid that contained our phage) and did dilutions of that lysate. We then plated the various dilutions and repeated the process with a plaque from the post dilute plate that still had plaques and repeated the process. We repeated this until there were plates that had consistent plaque morphologies (same size and turbidity). The hope with this is that we will have a sample that has only one type of phage

  1. What is meant by “non contaminant phage” and why would you want to purify the phage lysate to isolate the non contaminant phage? Why is the contaminant phage not wanted?

    1. By “non contaminant phage” I meant the phage that was not the O cluster. Since our lab had a large scale contamination of the O cluster phage, that phage was the contaminant phage. I wanted to purify the lysate to only contain the non contaminant phage because that is the phage we know where it is from. We don’t want the contaminant phage because it cannot be archived since its origin is unknown. And if the phage are not archived, they are not part of the SEA PHAGES database, and will not be available for phage therapy.

    1. In the purification process phage were collected from a plaque (by scraping the plaque) and made into a phage lysate by adding it to phage buffer. This lysate was then diluted into more buffer, and the dilutions were plated. Another phage lysate was made from a plaque on the most dilute plate, and the process was repeated until plates contained plaques with consistent morphology. The consistent morphology indicated that the plate likely only had one type of phage

    2. Phage were taken from a plaque and put into phage buffer to create a phage lysate. The lysate was then diluted using more phage buffer, and the dilutions were plated. A new lysate was then made using a plaque from the most dilute plate and the process was repeated until all the plaques had consistent morphology. The consistent morphology indicates that there is likely only one type of phage present in the lysate

  2. Do you think that these bacteriophages may potentially mutate in hosts and become dangerous to the people they are being used to treat bacteria in? would this be more or less dangerous than anti-biotic resistant bacteria?

    1. Viruses do mutate a lot so it is definitely likely they will mutate in the hosts. However it is currently unknown if any such mutations could cause them to harm the people they are being used to treat. Since they are bacteria specific viruses they will likely not be able to directly infect any eukaryotic cells, however it is possible that mutations they do have can cause unintended effects. Their mutations could cause them to kill of necessary bacteria, or interfere with other systems within the person. There are currently a lot of unknowns in phage therapy which is why it is not yet an FDA approved treatment outside of specific dire circumstances. Phage therapy is currently only used in people who have lethal antibiotic resistant bacterial infections so that the potential benefits of phage therapy outweigh the potential costs. However for more generalized treatment I don’t have enough knowledge to say whether or not it would be less dangerous then anti-biotic resistant bacteria, but if I had to guess I would say it would be less dangerous.

  3. Would you be able to possibly produce better plaques if you were to use a different media that you plate the phage on?

    1. I don’t know enough about what makes a specific media more effective, but I don’t think it would create much of a difference. By my understanding, the media’s main function is to feed and support the bacteria. And since the plaques are from the viruses killing the bacteria, I don’t think there would much of an effect as long as both can survive and move through the media. The virus’ effectiveness in killing the bacteria is the major contributor to what the plaques look like, so I don’t think a different media would change how the plaques look.

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