Lactiplantibacillus plantarum CECT 4528 for silage (1k20746) renewed until 19 November 2035: PFGE to CEN/TS 17697 and MRS agar to EN 15787; gentamicin MIC 64 and kanamycin MIC 256 mg/l — two dilutions above the cut-off, but no acquired resistance genes
Commission Implementing Regulation (EU) 2025/2175, published on 30 October 2025, renews until 19 November 2035 the authorisation of a preparation of Lactiplantibacillus plantarum CECT 4528 as a silage additive for all animal species and repeals in its entirety Regulation (EU) No 399/2014, which also covered Lactobacillus brevis DSM 23231 and DSMZ 16680 and Lactobacillus fermentum NCIMB 30169. In its opinion of 28 January 2025 EFSA found the additive still safe; in a test to OECD TG 492 it is not an eye irritant, but as a micro-organism it is a skin and respiratory sensitiser.
- Methods in the annex: identification in the additive — PFGE to CEN/TS 17697 or DNA sequencing; enumeration — spread plate or pour plate on MRS agar (EN 15787). The reference laboratory considered the conclusions from the previous authorisation still valid, so there was no new report. Additive: at least 2.5 × 10¹¹ CFU/g, minimum dose without other micro-organisms 1 × 10⁹ CFU/kg fresh material; skin and respiratory protection. Additive and feed labelled before 19 November 2026 — until stocks are exhausted.
- Data from the EFSA opinion: the preparation consists of cells with maltodextrin (approx. 25–35 % cells, 25–35 % cryoprotectant, 30–50 % carrier — maltodextrin or lactose); five batches 3.4 × 10¹¹ CFU/g on average (3.0–3.7 × 10¹¹). Enterobacteriaceae, yeasts and fungi < 100 CFU/g, Salmonella not detected in 25 g (six batches). Arsenic 0.023–0.038, cadmium 0.022–0.024, lead 0.010–0.024 mg/kg; mercury and aflatoxin M1 below the LOQ. No new data on dusting potential or stability — the data from the 2014 opinion apply.
- The strain was isolated from ensiled grass. Identification was confirmed by digital DNA–DNA hybridisation (dDDH) of the whole genome sequence: 89.4 % with the type strain L. plantarum DSM 20174T; no plasmids were detected. Microdilution: chloramphenicol 16 mg/l against a cut-off of 8 (one dilution — within the variation of the method), gentamicin 64 against 16 mg/l and kanamycin 256 against 64 mg/l. No resistance genes above the EFSA thresholds were found in the genome (ResFinder, NCBI), so this resistance was considered of no concern.
- User: in 2014, in the absence of data, EFSA assumed possible skin and eye irritation; now a test to OECD TG 492 showed that the additive is not an eye irritant (UN-GHS “No Category”). A literature search covering 2014–2024 (Web of Science and PubMed) returned no hits relevant to safety. Efficacy was not assessed, because the application did not change the conditions of authorisation.