Bacillus subtilis DSM 33862 with Lentilactobacillus buchneri as an acidity regulator for moist plant feed — counts on TSA (EN 15784) and MRS agar (EN 15787), identification by sequencing or PFGE (CEN/TS 17697)
Commission Implementing Regulation (EU) 2026/2074 of 17 September 2026, published on 18 September, authorises until 8 October 2036 a preparation of two strains for all animal species — only in chopped feed materials of plant origin with 64–87 % moisture, at a minimum of 1 × 10⁸ CFU/kg. The preparation contains at least 3.6 × 10¹¹ CFU/g in a 1:4 ratio.
- Methods: identification of both strains — DNA sequencing or pulsed-field gel electrophoresis PFGE (CEN/TS 17697); B. subtilis count — spread plate on tryptone soya agar (EN 15784), L. buchneri count — spread or pour plate on MRS agar (EN 15787).
- Five batches: B. subtilis 7.98 × 10¹⁰ CFU/g (specification ≥ 7.2 × 10¹⁰), L. buchneri 3.69 × 10¹¹ CFU/g (≥ 2.88 × 10¹¹), total 4.49 × 10¹¹ CFU/g. Bacillus cereus < 100 CFU/g, Salmonella absent in 25 g, dusting potential 5–15 mg/m³.
- Efficacy was supported by three in vitro studies: crushed oats (36.3 % DM), carrots (13.1 %) and sugar beet (18.4 %) incubated for 24 h at 25 °C in vacuum bags, three replicates each. With the preparation the pH fell to 4.83, 4.06 and 5.02 versus 5.95, 6.02 and 6.18 in the controls.
- The B. subtilis strain was identified by digital DNA–DNA hybridisation based on its whole genome sequence; it carries no acquired resistance genes (ResFinder, AMRFinderPlus), and a Vero cell test ruled out toxigenic potential. EFSA concluded that the preparation should be considered a skin and respiratory sensitiser.