Commission Implementing Regulation (EU) 2025/2491 of 10 December 2025 concerning the authorisation of a preparation of 25-hydroxycholecalciferol produced with Saccharomyces cerevisiae CBS 146008 as a feed additive for all animal species except poultry, pigs and ruminants

Official Journal EN of the European Union L series 2025/2491 11.12.2025 COMMISSION IMPLEMENTING REGULATION(EU) 2025/2491 of 10 December 2025 concerning the authorisation of a preparation of 25-hydroxycholecalciferol produced with Saccharomyces cerevisiaeCBS 146008 as a feed additive for all animal species except poultry, pigs and ruminants (Text with EEA relevance) THE EUROPEAN COMMISSION, Having regard to the Treaty on the Functioning of the European Union, Having regard to Regulation (EC) No 1831/2003 of the European Parliament and of the Council of 22 September 2003 on additives for use in animal nutrition(1), and in particular Article 9(2) thereof,

Whereas:

(1) Regulation (EC) No 1831/2003 provides for the authorisation of additives for use in animal nutrition and for the grounds and procedures for granting such authorisation.

(2) In accordance with Article 7 of Regulation (EC) No 1831/2003, an application was submitted for the authorisation of a preparation of 25-hydroxycholecalciferol produced with Saccharomyces cerevisiaeCBS 146008. That application was accompanied by the particulars and documents required under Article 7(3) of Regulation (EC) No 1831/2003.

(3) That application concerns the authorisation of a preparation of 25-hydroxycholecalciferol produced with Saccharomyces cerevisiae CBS 146008 as a feed additive for all animal species except poultry, pigs and ruminants, requesting that additive to be classified in the additive category ‘nutritional additives’ and in the functional group ‘vitamins, pro-vitamins and chemically well-defined substances having similar effect’.

(4) The European Food Safety Authority (‘the Authority’) concluded in its opinion of 20 May 2025(2) that, under the proposed conditions of use, the preparation of 25-hydroxycholecalciferol produced with Saccharomyces cerevisiaeCBS 146008 is safe for all animal species except poultry, pigs and ruminants, as well as for consumers and the environment. The Authority also concluded that the additive is not irritant to the skin or eyes. However, due to lack of information, no conclusion on its potential to be a skin sensitiser or on its effects on the respiratory system could be reached The Authority further concluded that the additive is efficacious in covering the target animals’ nutritional requirements. It did not consider that there is a need for specific requirements of post-market monitoring.

(5) The Reference Laboratory set up by Regulation (EC) No 1831/2003 considered that the conclusions and recommendations reached in a previous assessment concerning another application for the authorisation of the same additive and verified by the Authority in its opinion of 5 of July 2023(3) are valid and applicable for the current application. In accordance with Article 5(4), point (a), of Commission Regulation (EC) No 378/2005(4), an evaluation report of the Reference Laboratory is therefore not required.

(1) OJ L 268, 18.10.2003, p. 29, ELI: http://data.europa.eu/eli/reg/2003/1831/oj.

(2) EFSA Journal. 2025;23:e9479. https://doi.org/10.2903/j.efsa.2025.9479.

(3) EFSA Journal2023; 21(8):8168. https://doi.org/10.2903/j.efsa.2023.8168.

(4) Commission Regulation (EC) No 378/2005 of 4 March 2005 on detailed rules for the implementation of Regulation (EC) No 1831/2003 of the European Parliament and of the Council as regards the duties and tasks of the Community Reference Laboratory concerning applications for authorisations of feed additives (OJ L 59, 5.3.2005, p. 8, ELI: http://data.europa.eu/eli/reg/ 2005/378/oj).

ELI: http://data.europa.eu/eli/reg_impl/2025/2491/oj 1/4EN OJ L, 11.12.2025

(6) In view of the above, the Commission considers that a preparation of 25-hydroxycholecalciferol produced with Saccharomyces cerevisiae CBS 146008 satisfies the conditions provided for in Article 5 of Regulation (EC) No 1831/2003. Accordingly, the use of that preparation should be authorised for all animal species except poultry, pigs and ruminants. In addition, the Commission considers that appropriate protective measures should be taken to prevent adverse effects on the health of the users of the additive.

(7) The measures provided for in this Regulation are in accordance with the opinion of the Standing Committee on Plants, Animals, Food and Feed,

HAS ADOPTED THIS REGULATION:

Article 1 Authorisation The preparation specified in the Annex, belonging to the additive category ‘nutritional additives’ and to the functional group ‘vitamins, pro-vitamins and chemically well-defined substances having similar effect’, is authorised as an additive in animal nutrition, subject to the conditions laid down in that Annex.

Article 2 Entry into force This Regulation shall enter into force on the twentieth day following that of its publication in the Official Journal of the European Union.

This Regulation shall be binding in its entirety and directly applicable in all Member States.

Done at Brussels, 10 December 2025.

For the Commission The President Ursula VON DER LEYEN 2/4 ELI: http://data.europa.eu/eli/reg_impl/2025/2491/ojANNEX Minimum Maximum content content Identification Species or End of period Name of the Composition, chemical formula, Maximum number of the category of Other provisions of Additive description, analytical method. age mg active substance/kg of feed additive animal authorisation complete feed with a moisture content of 12 %

Category: Nutritional additives. Functional group: Vitamins, provitamins and chemically well-defined substances having similar effect

Sub classification: vitamin D 3a670a 25-Hydroxychole­ Additive composition Salmonids 0,800 1. The additive shall be 31 December calciferol incorporated in feedingstuffs 2035 Preparation with a maximum content Finfish species 0,075 via the use of a premixture.

of 1,25 % of other than 25-hydroxycholecalciferol salmonids 2. In the directions for use of the additive and premixtures, the Solid form Ornamental 0,075 storage conditions and the fish stability to heat treatment Characterisation of the active substance shall be indicated.

Other species 0,050 25-Hydroxycholecalciferol. Its and categories 3. Maximum content of the precursor of animals combination of compound,5,7,24-cholestatrienol, is except pigs, 25-hydroxycholecalciferol produced with Saccharomyces cerevisiae poultry and with cholecalciferol (vitamin CBS 146008. After extraction, the ruminants D ) per kg of complete 3

precursor is converted chemically to feedingstuff:

25-hydroxy-pro-vitamin D , which is 3 — ≤ 1,500 mg(2) further transformed photochemically (equivalent to 60 000 IU to 25-hydroxycholecalciferol. of cholecalciferol) for

Chemical formula: C H O •H O salmonids, 27 44 2 2

CAS number: 63283-36-3 — ≤ 0,075 mg(2) (equivalent to 3 000 IU

Purity criteria: of cholecalciferol) for — 25-hydroxycholecalciferol > 94 %; fish species other than — other sterol derivatives ≤ 1 % each; salmonids and for — erythrosine < 5 mg/kg. ornamental fish,

ELI:

http://data.europa.eu/eli/reg_impl/2025/2491/oj 3/4 OJ L,

11.12.2025 ENMinimum Maximum content content Identification Species or End of period Name of the Composition, chemical formula, Maximum number of the category of Other provisions of Additive description, analytical method. age mg active substance/kg of feed additive animal authorisation complete feed with a moisture content of 12 % Analytical method(1) — ≤ 0,050 mg(2) (equivalent to 2 000 IU For the determination of of cholecalciferol) for 25-hydroxycholecalciferol in the feed other categories of

additive: Ultra Performance Liquid animals except poultry, Chromatography coupled to pigs and ruminants. spectrophotometric detection (UPLC-UV). 4. For users of the additive and premixtures, feed business For the determination of operators shall establish 25-hydroxycholecalciferol in operational procedures and

premixtures: High Performance Liquid organisational measures to Chromatography coupled to address the potential risks spectrophotometric detection resulting from their use. (HPLC-UV).

Where those risks cannot be For the determination of eliminated by such 25-hydroxycholecalciferol in procedures and measures, the compound feed and in low additive and premixtures

concentrated premixtures: High shall be used with personal Performance Liquid Chromatography breathing and skin protective coupled to tandem mass spectrometry equipment. (HPLC-MS/MS).

(1) Details of the analytical methods are available at the following address of the Reference Laboratory: https://joint-research-centre.ec.europa.eu/eurl-fa-eurl-feed-additives/eurl-fa-authorisation/eurl-fa- evaluation-reports_en.

(2) 40 IU cholecalciferol (vitamin D3) = 0,001 mg cholecalciferol (vitamin D).

3 4/4

ELI:

http://data.europa.eu/eli/reg_impl/2025/2491/oj EN OJ L,

11.12.2025