Assessment of two different methods for sampling and detection of Dichelobacter nodosus and Fusobacterium necrophorum in dairy cows in Eastern Slovakia.

The aim of our study was to find the most appropriate way of sample collection from cattle feet as well as to assess simple and effective sample processing, including DNA extraction for reliable diagnosis of bacteria Dichelobacter nodosus and Fusobacterium necrophorum. 11 clinically healthy cows were included in the study, from which swabbing samples (2 types: surface swab and deep swab) were taken. Two isolation methods were used for DNA extraction: 1. freezing and boiling the samples, 2. commercial kit (Roche). PCR analysis of the samples has not shown any variations in the detection ratio of D. nodosus and F. necrophorum at different swabbing methods. The highest sensitivity of the detection of both bacteria was reached with a cultivation of samples in AB with subsequent extraction of DNA with freezing and boiling. The cultivation in anaerobic broth resulted in the detection rate of D. nodosus and F. necrophorum in over 95% and 27%, respectively. To conclude, the simple 'surface' swab is sufficient to detect studied pathogens, the most appropriate method of DNA extraction has proven to be freezing and boiling of the sample. [ABSTRACT FROM AUTHOR]