Last week was a very busy week for me in the laboratory and on the farms. Monday was spent screening faecal samples from blackface mountain ewes from county Galway. These ewes contained high amounts of both rumen and liver fluke, this is understandable as they come from a wet area which could give rise to the intermediate host, and the owners of mountain sheep would be less likely to regular dose their sheep against fluke. Tuesday was my first day collecting samples on a farm. Michael Fagan who is the farm manager brought me two Teagasc farms Tuohy’s and Newford, Tuohy’s is a rented farm where they rear their steer (castrated) bull claves for dawn meats and Newford is where they have their suckler herd. Michael brought me out to take samples from Tuohy’s because he felt that these bullocks were not thriving as well as before Christmas and suspected a fluke infection. I collected 11 samples, six from tagged calves and five from ground samples. After Tuohy’s we headed for Newford where I collected 18 faecal samples from cows around the farm. One cow on this farm was quite ill so samples had previously been taken from this cow and both rumen and liver fluke eggs were identified. I collected samples from this cow and 16 others in the hope of identifying rumen fluke eggs. These 29 samples were stored in the fridge overnight. On Wednesday and Thursday, I prepared both Tuohy’s and Newford samples through the sedimentation method and the addition of the methylene blue and began to screen the slides. On Friday I continued to screen these samples and recorded rumen fluke eggs in only one of the samples from the Tuohy farm, next week I will run PCR on all 11 samples to see if there is rumen fluke DNA in the samples. If there is RF DNA in the samples where no eggs were found, then the animal is infected with the immature stage which is causing harm to the animal but not shedding eggs. Next week I will continue screening, running PCR and finish the corrections for my Lit review.
My week was mostly spent in NUIG with my course coordinator Galina
and my lab partner, here I learned how to carry out extraction of DNA from a
Haemonchus contortus. This is the parasite my partner will be carrying out his master’s
project on. Post extraction I learned how to perform washing and elution of the
DNA extracted from the worm and then we performed PCR.
This was my first time to carry out this laboratory work and
it worked successfully, we were also shown how to perform qPCR on our samples.
However, we could not make a quantitative PCR as we do not have other samples
of DNA present to compare the levels in the sample, because we extracted the
DNA form the worm and not the eggs of the sample.
Wednesday the 6th
of February marked my first day with Teagasc in Athenry Co, Galway. Upon
arrival I received my log in details for the teagasc website and email along
with my security pass for access to all rooms and labs in the department. To
commence research and lab work I presented my completed training certificates
for; laboratory health and safety training, Teagasc safety course and a manual
handling training course.
Padraig O’Boyle, my
project supervisor gave me a tour of the department and showed me how to access
the Teagasc document management system (DMS) which I will need to become
accustomed with before starting my lab work and write up. After I had acquired
the general basics I began to print research papers and standard operating
procedures which are related to my project and continued to study the papers
which Galina had sent me.
Thursday the 7th, I
continued reading research papers and learning SOP’s under Padraig’s guidance,
I also formulated a list of equipment and materials needed for carrying out my
laboratory work based off the 2016 paper on identifying and studying P.
On Thursday evening
Padraig and I carried out a FECPAK test for round worms in the lab which is a
sedimentation procedure used to identify parasites in the faeces of the
animals. This was my first time in the lab and using a SOP which I will be
using later for my own research.
On Friday the 8th
of February I scheduled my manual handling practical training and carried out
the FECPAK experiment for liver fluke and rumen fluke which is an extension of
the work I did on Thursday which involves further sedimentation and using
methylene blue to help identify the parasite under a microscope.