Bacillus velezensis RTI301 approved as a low-risk active substance from 12 February 2025 to 12 February 2040, together with Bacillus subtilis RTI477 (2025/106) from a shared product against soil-borne diseases; EFSA — strain identification with PCR primers and restriction analysis, but without demonstrating the absence of surfactin C and subtilisin in the technical material
Commission Implementing Regulation (EU) 2025/109, published on 23 January 2025, approves the new active substance Bacillus velezensis strain RTI301 as a low-risk substance, because no multiple resistance to antimicrobials used in human or veterinary medicine was demonstrated at strain level. The applicant was FMC Agricultural Solutions A/S, the rapporteur the Netherlands, and the EFSA conclusions date from 31 July 2024. On the same day Implementing Regulation 2025/106 approved, on the same terms and dates, Bacillus subtilis strain RTI477 — the second strain in the representative product.
- Annex: Bacillus velezensis RTI301, no relevant impurities, approval from 12 February 2025 to 12 February 2040; Member States must pay attention to the protection of operators and workers (microorganisms as potential sensitisers, personal protective equipment in the conditions of use) and to production quality control in line with the microbial contaminant limits of OECD Issue Paper No 65. The strain is deposited at DSMZ (DSM 32473) and ATCC (PTA-121165); it was isolated in 2013 from the rhizosphere of a vineyard on Long Island, New York. The Polish version of the act gives the expiry date in Annex II (the entry in Part D of the list) as “12 February 2025”; Annex I and the English version give 12 February 2040.
- Identification and methods (EFSA): species assignment — morphology and multilocus sequence analysis; strain-specific PCR primers and restriction enzyme analysis, developed from the whole genome sequence, distinguish the strain from other species and closely related strains. Content in the technical material 1.0 × 10¹¹–1.0 × 10¹² CFU/g; methods exist for determining the microorganism and microbial contaminants (below the limits). Monitoring methods were not required because no residue definitions were set. The product is stable for 24 months at 25 °C.
- Metabolites: EFSA considered subtilisin and surfactin C of potential concern for health and requires analytical demonstration that they are absent from the technical material (for subtilisin — below the threshold for classification as a sensitiser); the rapporteur considered surfactin C of no toxicological concern. Iturin A was considered unlikely on the basis of the genome. The strain has QPS status with qualifications — absence of toxigenic potential was confirmed by a negative Vero cell assay and genome analysis; in a phenotypic test it was susceptible to all 8 antibiotics tested. Recommended label phrase: ‘Contains B. velezensis strain RTI301. Microorganisms may have the potential to provoke sensitising reactions’.
- The representative product ‘F4034-5’ contains 9.0 × 10⁹–9.0 × 10¹⁰ CFU/mL of B. velezensis RTI301 (90 g/L) and 3.0 × 10⁹–3.0 × 10¹⁰ CFU/mL of B. subtilis RTI477 (30 g/L); it controls, among others, Rhizoctonia solani, Pythium spp., Phytophthora capsici, Sclerotinia sclerotiorum, Phoma lingam, Plasmodiophora brassicae and Fusarium spp. in potatoes, solanaceous and cucurbit vegetables, lettuce and in seed treatment of maize, sunflower, sugar beet and winter oilseed rape. EFSA did not finalise the non-dietary exposure assessment, the consumer risk assessment and the assessment for many groups of organisms; the strain meets none of the five criteria for inclusion in Annex IV to Regulation 396/2005.