ELI: http://data.europa.eu/eli/reg_impl/2024/764/oj 1/4COMMISSION IMPLEMENTING REGUL ATION (EU) 2024/764 
of 29 Febr uary 2024
concer ning the author isation of a preparation of Bacillus subtilis CNCM I-4606, CNCM I-5043 and 
CNCM I-4607 and Lactococcus lactis CNCM I-4609 as a feed additiv e for all animal species 
(Text with EEA relevance) 
THE EUR OPEAN COMMISSION,
Havi ng regar d to the Treaty on the Functioning of the European Union,
Havi ng rega rd to Regulation (EC) No 1831/2003 of the European Parliament and of the Council of 22 Sept ember 2003 on 
additives for use in animal nutr ition (1), and in particular Article 9(2) thereof,
Whereas:
(1) Regulation (EC) No 1831/2003 provid es for the author isation of additives for use in animal nutr ition and for the 
grounds and procedures for granting such an author isation.
(2) In accordance with Article 7 of Regulation (EC) No 1831/2003, an application was submitted for the author isation 
of a preparation of Bacillus subtilis CNCM I-4606, CNCM I-5043 and CNCM I-4607 and Lactococcus lactis CNCM 
I-4609. That application was accompan ied by the particulars and documents required under Article 7(3) of 
Regulation (EC) No 1831/2003.
(3) The application concer ns the author isation of the preparation of Bacillus subtilis CNCM I-4606, CNCM I-5043 and 
CNCM I-4607 and Lactococcus lactis CNCM I-4609 as a feed additive for all animal species, requesting that additive 
to be classifie d in the categor y ‘technological additives’ and in the functional group ‘hygiene condition enhancers’.
(4) The European Food Safety Author ity (‘the Author ity’) concluded in its opinions of 30 Sept ember 2021 (2)and of 
1 Febr uary 2023 (3)that, under the proposed conditions of use, which exclude the use of the additive in 
commercial premixtures containing trace elements or preser vatives, the preparation of Bacillus subtilis CNCM 
I-4606, CNCM I-5043 and CNCM I-4607 and Lactococcus lactis CNCM I-4609 is safe for the target species, 
consumers and the environment. It also concluded that the preparation is not irritant to skin or eyes or a skin 
sensitiser but that, given the proteinaceous nature of the active agent, it should be considered a respirato ry 
sensitiser . The Author ity further concluded that the additive at the minimum proposed inclusion level showed 
poten tial to reduce Salmonella Typhimur ium growth in feed with high moisture cont ent (60–90 % moisture) and 
note d that the use of this additive cannot be considered as a replacement for the standard hygiene farming 
conditions. The Author ity also verified the repor t on the method of analysis of the feed additive in feed submitted 
by the Reference Laboratory set up by Regulation (EC) No 1831/2003.
(5) In view of the above, the Commission considers that the preparation of Bacillus subtilis CNCM I-4606, CNCM I-5043 
and CNCM I-4607 and Lactococcus lactis CNCM I-4609 satisfi es the conditions provided for in Article 5 of Regulation 
(EC) No 1831/2003. According ly, the use of that preparation should be author ised. In addition, the Commission 
considers that appropr iate prot ective measures should be taken to prevent adverse effects on the health of the users 
of that additive.
(6) The measures provid ed for in this Regulation are in accordance with the opinion of the Standing Committ ee on 
Plants, Animals, Food and Feed,
(1)OJ L 268, 18.10.2003, p. 29.
(2)EFSA Journal 2021;19(11):6907.
(3)EFSA Journal 2023;21(3):7871.
Offi cial Jour nal  
of the European Union EN 
L series    
2024/764  1.3.2024ELI: http://data.europa.eu/eli/reg_im pl/2024/764/oj 2/4HAS ADOPTED THIS REGUL ATION:
Article 1
The preparation specifi ed in the Annex, belonging to the additive catego ry ‘techn ological additives’ and to the functional 
group ‘hygiene condition enhancers ’, is author ised as an additive in animal nutr ition, subject to the conditions laid down 
in that Annex.
Article 2
This Regulation shall enter into force on the twentieth day followi ng 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, 29 Febr uary 2024 .
For the Commission
The President
Ursula VON DER LEYENOJ L, 1.3.2024 EN  ELI: http://data.europa.eu/eli/reg_impl/2024/764/oj 3/4ANNEX 
Identifi -
cation 
number 
of the 
additiveAdditiveComposition, chemical formula, descr iption, 
analytical methodSpecies or 
categ ory of 
animalMaximum 
ageMinimum conten tMaximum 
conten t
Other provisi onsEnd of period of 
author isation
CFU/L of compl ete feed ingstuff
Categor y: technological additiv es. Functional group: hygiene condition enhancers (reduction of the contamination by Salmonella Typhimur ium)
1n01 Bacillus subtilis 
CNCM I-4606, 
CNCM I-5043 
and CNCM 
I-4607 and 
Lactococcus lactis 
CNCM I-4609Additive composition
Preparation of Bacillus subtilis CNCM I-4606, 
CNCM I-5043 and CNCM I-4607 and 
Lactococcus lactis CNCM I-4609 containing a 
minimum of:
— 2 × 109 CFU/g in tota l for the three Bacillus 
subtilis strains (in ratio 1:1:1);
— 2 × 109 CFU/g for the Lactococcus lactis.All animal 
species- 1 × 109 of Bacillus 
subtilis CNCM I- 
4606, 4607, 
and 5043 (in 
ratio 1:1:1)
1 × 109 of  
Lactococcus lactis 
CNCM I-4609- 1. In the directions for use of the 
additive and premixtures, the 
stora ge conditions and stabi ­
lity to heat treatment shall be 
indicated .
2. The additive shall only be used 
in mash compound feed or in 
solid feed mat erials, intended 
for the preparation of feed with 
a moisture content of 60-90 %.
3. The additive shall not be used 
in premixtures containing 
trace elements or preser va­
tives.
4. On the label of the additive and 
premixtures, the following 
shall be indicated : ‘The additive 
1n01 is intended for the reduc ­
tion of the contamination by 
Salmonella Typhimur ium. It 
cannot be considered as a re­
placement for the standard hy­
giene farming conditions.’21 March 2034
Char acter isation of the active substance
Viable spores of Bacillus subtilis strains CNCM 
I-4606, CNCM I-5043 and CNCM I-4607 and 
live cells of Lactococcus lactis CNCM I-4609
Analytical method (1)
Enumeration in the feed additive, premixtures 
and compound feeds of the total Bacillus subtilis 
CNCM I-4606, CNCM I-5043 and CNCM 
I-4607:
— Spread plate method on tryptone soya agar 
(EN 15784);
Enumeration in the feed additive, premixtures 
and compo und feeds of the Lactococcus lactis 
CNCM I-4609:
— Poured plate method on MRS agar (EN 
15214).OJ L, 1.3.2024 EN  ELI: http://data.europa.eu/eli/reg_im pl/2024/764/oj 4/4
5. For users of the additive and 
premixtures, feed business op­
erators shall establish opera ­
tional procedures and organi ­
sational measures to address 
poten tial risks resulting from 
their use. Where those risks 
cannot be eliminated by such 
procedures and measures, the 
additive and premixtures shall 
be used with personal breath ­
ing prote ctive equipment.Identif ication of Bacillus subtilis CNCM I-4606, 
CNCM I-5043 and CNCM I-4607 and of the 
Lactococcus lactis CNCM I-4609:
Pulsed Field Gel Electrophoresis 
(PFGE) CEN/TS 17697 or DNA sequencing 
methods
(1)Details of the analytical methods are available at the following address of the Reference Laborato ry: https://joint-research-c entre.ec.europa.eu/eurl-fa-eurl-f eed-additives/eurl-fa-author isation/eurl-f a-evaluation- 
repor ts_enOJ L, 1.3.2024 EN  