E swabs 480ce

Samples (n = 2,126) were collected from 10 sheep farms situated within Nottingham, Derby and Northampton from animals with varying disease states. Sterile nylon flock swabs (E-swabs 480CE, Copan U.S.A.) were used for the collection of samples from the interdigital space of sheep and stored in liquid Amies solution at 5°C overnight. The swabs were inoculated onto Hoof Agar plates containing 4% w/v Bacto Eugon agar (BD, U.S.A.), 0.5% w/v Difco Yeast Extract (BD, U.S.A.), 1.5% w/v BBL Beef Extract (BD, U.S.A.), 1% Sodium Chloride and 6.6% w/v ovine hoof powder (Parker et al., 2005 (link)) and incubated anaerobically at 37°C. After 7 days plates were visually checked for putative D. nodosus and if present sub-cultured onto reduced agar (2%), hoof agar plates and incubated anaerobically at 37°C. Pure colonies were collected from plates in sterile PBS, washed by centrifugation and resuspended in molecular biology grade water (ThermoFisher, U.K.).

Sheep were assessed after slaughter for foot health. Any individual animals showing signs of footrot were selected for sample collection. Debris was removed from all feet, and the feet were cleaned using purified water. Sterile nylon flock swabs (E-swabs 480CE; Copan, USA) were taken from the interdigital space and stored in liquid Amies medium at 5 °C overnight. The foot was then washed with a chlorohexidine solution (National Veterinary Services, UK). Any hair was removed from the feet with scissors prior to the collection of an 8-mm biopsy specimen using a punch (National Veterinary Services, UK). Biopsy specimens were placed in RNAlater (Sigma-Aldrich, UK) and stored at 5 °C overnight before being frozen at −80 °C. This study was reviewed and approved by the University of Nottingham School of Veterinary Medicine and Science ethical review committee (ERN 1144 140506 [non-ASPA]).