Can someone take a look and confirm whether what I isolated is indeed C. jejuni? I haven't got any primers yet, so I can't confirm by PCR, but the culture is gram negative, catalaze +, oxidase +. The colonies are kind of large and flat and do not look like classical translucent and shapeless water droplets. Also, I have read that under stress the bacilli may appear coccoid. Mine didn't grow under microaerophillic conditions, I just grew them in a plastic bag at 42C for 48 hours. Isolated from raw chicken breasts from the supermarket. Just getting ready for my next project and I'd appreciate a feedback. Thanks!
1- I wonder if the culturing procdure is done under microaerophilic conditions?
2- Yes is true under stress When exposed to atmospheric oxygen, C. jejuni is able to change into a coccal form.
3- You can read this article , may help you:Morphology heterogeneity within a Campylobacter jejuni helical population: The use of calcofluor white to generate rod-shaped C. jejuni 81-176 clones and the genetic determinants responsible for differences in morphology within 11168 strains. Emilisa Frirdich, Jacob Biboy, Steven Huynh, Craig T. Parker, Waldemar Vollmer, Erin C. Gaynor.2017.Molecular Microbiology.
4- The genome of C. jejuni strain NCTC11168 was published in 2000.
with best regards
Hi,
Thanks for the comment! No, I said that I did NOT grow them under MA conditions-haven't gotten the shipment of pouches yet.
So, that must be it then. Have to check them on motility assay as well.
They were grown on Preston agar for 48 hours at 42C, but not under microaerophillic conditions. Just in a plastic bag infused with C02.
It is possible that after infusing the CO2, the concentration of oxygen has decreased so that the atmosphere becomes microaerophilic (Less than 5% oxygen).
The colonies on CCDA agar are gray and creamy and The bacterial morphology seen in your photograph is compatible with Campylobacter (Look at the attached file)
Thank you. Although the attached photograph looks a lot better :)
I ordered motility media and will check if they are motile-that would be another way to confirm- and tomorrow I am going to order primers for species-specific ID.
You can see the motility in a wet mount using a colony suspension in physiological saline solution
Campylobacter ieiuni characterized by wing like shape and may under stress change to coccal form. make suspention in brian heart infusion broth and incubated it for at least 3 days then filtrate the culture by using 0.45 filter paper directly on brain heart infusion agar supplemented with blood. if it campylobacter, will pass throught filter paper because small size.
Good luck
thanks, I think PCR sounds more definitive. A lof of things can pass through the filter paper :)
you can use only 0.45 millipore and then confirmed by Hi Campylobacter Latex test kit
Actually, it turned out campy as per specific 16S RNA PCR.
Repeating the PCR once again. Using E.coli for specificity and will see what we get.
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