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ABSTRACTThe aim of this study was to determine the prevalence of bacteria Dichelobacter nodosus(D. nodosus) and Fusobacterium necrophorum(F. necrophorum) in non-lame dairy cows on eight Slovak dairy herds. In total, 1,631 Holstein Friesian dairy cows were included in the study. Information of the cows was collected from on-farm software included cow ID, lactation number (heifer/cow), year milk yield (kg), and DIM on collection date. The PCR method detected D. nodosusand F. necrophorumon the feet of 1,394 (85.5%) and 373 (22.1%) dairy cows, respectively (p < 0.05). No dairy farm could be found without positive samples for D. nodosusand the majority of the farms were free or had very low prevalence of F. necrophorum(0–9.1%). Statistical difference for the prevalence of D. nodosusand F. necrophorumin different Slovak regions was detected for both strains. Logistic regression revealed an association between both D. nodosusand F. necrophorumidentification and annual milk production (odds ratios = 0.69 and 0.32, respectively). Our data demonstrates that D. nodosusis present on the feet of almost every dairy cow what means a potential risk for cattle to develop foot rot. Furthermore, both bacteria are associated with decreased annual milk production.

ABSTRACTThe 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 nodosusand 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. nodosusand F. necrophorumat 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. nodosusand F. necrophorumin 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.