Cows (n=40) on a Danish dairy herd were randomly selected on the basis of a uterine score indicating that the cows had uterine pathology. Uterine fluid was aspirated and if necessary the uterus was flushed with 30 ml sterile saline solution in order to retrieve uterine material. The fluid was placed in RNAlater. An endometrial biopsy was retrieved and the tissue placed in RNAlater. The cows were sampled on days 5-11 (week 1), days 26-32 (week 4), and on days 47-53 (week 7). This sampling schedule provided an opportunity to follow the development of any infection, and the combination of biopsy and uterine flush samples offered insights into whether tissue-invasive bacteria were present. The DNA was extracted with the Maxwell 16 LEV Blood kit (Promega), the 16S rRNA PCR was performed with primers targeting the V2 region, and the 454 next generation sequencing was performed by GATC.
Previous papers based on culturing of endometrial swabs or biopsies point to Escherichia coli, Trueperella (Archanobacterium) pyogenes, and Fusobacterium necrophorum as the most likely pathogens, although some of them also seem to be present in healthy animals. We expect to find these bacteria in the samples from the diseased animals, and perhaps the detailed data from the sequencing will also reveal hitherto undiscovered pathogens.
|Number of pages||1|
|Publication status||Published - 2012|
|Event||14th International Symposium on Microbial Ecology: The Power of the Small - Bella Center, Copenhagen, Denmark|
Duration: 19 Aug 2012 → 24 Aug 2012
Conference number: 14
|Conference||14th International Symposium on Microbial Ecology|
|Period||19/08/2012 → 24/08/2012|