Enterococcus lactis NCIMB 10415

In the current application authorisation is sought under Article 10(2) for Enterococcus faecium NCIMB 10415, under the category/functional group 4(b), ''zootechnical additives/gut flora stabilisers'', according to the classification system of Annex I of Regulation (EC) No 1831/2003. The feed additive will be marketed as micro-encapsulated or granulated form with concentrations of Enterococcus faecium NCIMB 10415: ranging from 5x10⁹ to 3.5x10¹⁰ CFU/g. Specifically, authorisation is sought for the use of the feed additive for pigs for fattening¹, piglets (suckling and weaned)¹, sows¹, cats² and dogs². The feed additive is intended to be used either via premixtures or incorporated directly into the feedingstuffs. The Applicant proposed the following content of Enterococcus faecium NCIMB 10415 in complete feedingstuffs: - for dogs and cats a minimum content of (4.5 and 5) x10⁶ CFU/kg; - for pigs and piglets a content range of (3.5 – 10) x10⁸ CFU/kg; and - for sows a content range of (0.7 – 1.25) x10⁹ CFU/kg.

For the enumeration of Enterococcus faecium NCIMB 10415 in the feed additive, premixtures and feedingstuffs the Applicant proposed two single laboratory validated pour plate/spread plate methods. The EURL identified instead the internationally recognised ring-trial validated spread plate CEN method (EN 15788), using Bile Esculin Azide Agar, for the enumeration of Enterococcus spp. The performance characteristics of the CEN method reported after logarithmic transformation (CFU) are:

• a repeatability standard deviation (s_r) ranging from 0.12 to 0.2 log₁₀ CFU/g,
• a reproducibility standard deviation (s_R) ranging from 0.23 to 0.41 log₁₀ CFU/g; and
• a limit of detection (LOD) of 1x10⁵ CFU/kg feedingstuffs.

Based on the performance characteristics presented, the EURL recommends for official control, the CEN method (EN 15788) for the enumeration of Enterococcus faecium NCIMB 10415 in feed additive and feedingstuffs.

Molecular methods were used by the Applicant to identify the active agent in the feed additive. The EURL recommends instead for official control Pulsed Field Gel Electrophoresis (PFGE), a generally recognised standard methodology for microbial identification. 

Further testing or validation of the methods to be performed through the consortium of National Reference Laboratories as specified by article 10 (Commission Regulation (EC) No 378/2005) is not considered necessary.

¹FAD-2010-0269; ² FAD-2010-0294

Recommended Method

as of:

- Commission Implementing Regulation (EU) 2024/2394

- Commission Implementing Regulation (EU) 2024/1161

Identification:

• DNA sequencing methods or Pulsed Field Gel Electrophoresis (PFGE) – CEN/TS 17697¹

Enumeration in the feed additive, premixtures, compound feed and water: 

Spread plate method using bile esculin azide agar (EN 15788)³

³ Text according to Commission Implementing Regulation (EU) 2024/1161