COMMISSION IMPLEMENTING REGUL ATION (EU) 2021/969 
of 16 June 2021
concer ning the author isation of L-threonine produced by Escherichia coli CGMCC 13325 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 author isation.
(2) In accordance with Article 7 of Regulation (EC) No 1831/2003, an application was submitted for the author isation 
of L-threonin e produced by Escher ichia coli CGMCC 13325 as a feed additive for use in feed for all animal species. 
That application was accompan ied by the particulars and documents required under Article 7(3) of that Regulation.
(3) The application concer ns the author isation of L-threonine produced by Escher ichia coli CGMCC 13325 as a feed 
additive for all animal species to be classifie d in the additive categor y ‘nutr itional additives ’.
(4) The European Food Safet y Author ity (‘the Auth ority’) concluded in its opinion of 18 Novem ber 2020 (2)that, under 
the proposed conditions of use, L-threonine produced by Escher ichia coli CGMCC 13325 does not have an adverse 
effect on animal health, consumer health or the environment. It could not conclude on the potential of L-threonine 
produced by Escher ichia coli CGMCC 13325 to be a skin sensitiser and irritant to the skin and eyes, and stated an 
inhalation risk to endoto xins for the users of the additive. Theref ore, appropr iate protective measures should be 
taken to prevent adverse effects on human health, in particular as regar ds the users of the additive. The Author ity 
also concluded that the additive is an efficacious source of the amino acid L-threonin e for all animal species and 
that in order to be as effic acious in ruminants as in non-r uminant species, the additive needs to be prote cted against 
degradation in the rumen. The Auth ority does not consider that there is a need for specific requirements of post- 
mark et monitoring. It 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) The assessment of L-thr eonine produced by Escher ichia coli CGMCC 13325 shows that the conditions for 
author isation, as provided for in Article 5 of Regulation (EC) No 1831/2003, are satisfied. According ly, the use of 
this additive should be author ised as specifi ed in the Annex to this Regulation.
(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,
HAS ADOPTED THIS REGUL ATION:
Article 1
The substance specifi ed in the Annex, belonging to the additive categor y ‘nutr itional additives ’and to the functional group 
‘amino acids, their salts and analogues ’, is author ised as an additive in animal nutr ition, subject to the conditions laid down 
in that Annex.
(1)OJ L 268, 18.10.2003, p. 29.
(2)EFSA Journal 2020;18(12):6332.EN Offi cial Jour nal of the European Union 17.6.2021                                                                                                                                         L 214/49  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, 16 June 2021 .
For the Commission
The President
Ursula VON DER LEYENEN Offi cial Jour nal of the European Union L 214/50                                                                                                                                         17.6.2021  ANNEX 
Identif ication 
number of the 
additiv eName of the 
holder of 
author isa-
tionAdditiv eComposition, chemical formula, 
descr iption, analytical method.Species or 
categor y of 
animalMaximum 
ageMinimum 
contentMaximum 
content
Other provisionsEnd of period 
of 
author isationmg/kg of complete feed 
with a moistu re content of 
12 %
Categor y of nutr itional additiv es. Functional group: amino acids, their salts and analogues.
3c411 - L-threonine Additiv e composition:
Powder with a minimum of 98 %
L-thr eonine and a maximum 
moisture conte nt of 1 %All species - - - 1.L-threonine may be placed on 
the mark et and used as an ad­
ditive consisting of a prepara ­
tion.
2.L-threonine may be used via 
water for drinking.
3.The labelling of the additive 
shall indicate the moisture 
content.
4.The endoto xin conte nt of the 
additive and its dusting po­
tential shall ensure a maximal 
endoto xin exposure of 1 600
IU endoto xins/m3 air(2).
5.The labelling of the additive 
and premixtures shall indi ­
cate the follo wing:
‘The supplementation with 
L-threonine, in particular via 
water for drinking, should take 
into account all essential and 
conditionally essential amino 
acids in order to avoid 
imbalances. ’7.7.2031
Character isation of the activ e 
subst ance:
L-thr eonine produced by 
fermentation with Escher ichia coli 
CGMCC 13325
Chemical formula: C4H9NO 3
CAS Number: 72-19-5.
Analytical methods (1):
For the determination of 
L-thr eonine in the feed additive:
— Food Chemical Codex 
‘L-threonine monograph ’, 
and
—ion exchange chromat ogra ­
phy coupled with post-col ­
umn derivatisation and opti­
cal detec tion (IEC- VIS/FLD) – 
EN ISO 17180.EN Offi cial Jour nal of the European Union 17.6.2021                                                                                                                                         L 214/51  6.For users of the additive and 
premixtures, feed business 
operators shall establish op­
erational procedures and or­
ganisational measures to ad­
dress the poten tial risks by 
inhalation, eye or dermal con­
tact. Where those risks can­
not be eliminated or reduced 
to a minimum by such proce ­
dures and measures, the addi ­
tive and premixtures shall be 
used with appropr iate perso ­
nal prote ctive equipment, in­
cluding breathing, skin and 
eye protection.For the determination of 
threonine in premixtures:
—ion exchange chromat ogra ­
phy coupled with post-col ­
umn derivatisation and opti­
cal detec tion (IEC- VIS/FLD) – 
EN ISO 17180, and
—ion exchange chromat ogra ­
phy coupled with post-col ­
umn derivatisation and opti­
cal detection (IEC- VIS), 
Commission Regulation (EC) 
No 152/2009 (Annex III, F).
For the determination of 
threonine in compo und feed and 
feed mater ials:
—ion exchange chromat ogra ­
phy coupled with post-col ­
umn derivatisation and opti­
cal detection (IEC- VIS): 
Commission Regulation (EC) 
No 152/2009 (Annex III, F).
For the determination of 
threonine in wate r:
—ion exchange chromat ogra ­
phy coupled with post-col ­
umn derivatisation and opti­
cal detection (IEC- VIS/FLD).
(1)Details of the analytical methods are avail able at the follo wing address of the Reference Laboratory : https://ec.europa.eu/jrc/en/eurl/feed-additives/evaluation-repor ts
(2)Exposure calculated based on the endoto xin level and the dusting potential of the additive according to the method used by EFSA (EFSA Journal 2015;13(2):4015); analytical method: European Phar macopoeia 
2.6.14. (bacterial endoto xins).EN Offi cial Jour nal of the European Union L 214/52                                                                                                                                         17.6.2021