Lentilactobacillus buchneri DSM 19455

In the current application authorisation is sought under Article 4(1) for Lactobacillus kefiri BIO 94 IFA 94 – DSM 19455¹, under the category/functional group 1(k), ''technological additives/silage additives'', according to the classification system of Annex I of Regulation (EC) No 1831/2003. Specifically, authorisation is sought for the use of the feed additive for all animal species and categories. The feed additive is to be placed on the market as a powder, containing a minimum concentration of 1x10¹⁰ CFU/g of Lactobacillus kefiri BIO 94 IFA 94 – DSM 19455. It is intended to be mixed directly into silage or suspended in water and sprayed on silage with a minimum concentration of 5x10⁷ CFU/kg fresh forage. 

For enumeration of Lactobacillus kefiri BIO 94 IFA 94 – DSM 19455 in feed additive, the Applicant submitted the ring-trial validated spread plate CEN method (EN 15787), using MRS agar. The performance characteristics of the method reported after logarithmic transformation are:

- a standard deviation for repeatability (S_r) of 0.24 log₁₀ CFU/g;

- a standard deviation for reproducibility (S_R) ranging from 0.29 to 0.38 log₁₀ CFU/g; and

- a limit of detection (LOD) of 10⁵ CFU/kg feedingstuffs.

Based on the performances characteristics presented, the EURL recommends for official control, the CEN method (EN 15787) for the determination of Lactobacillus kefiri BIO 94 IFA 94 – DSM 19455 in the feed additive per se.

The Applicant did not provide any experimental method or data for the determination of Lactobacillus kefiri BIO 94 IFA 94 – DSM 19455 in silage. Furthermore, the unambiguous determination of the content of Lactobacillus kefiri BIO 94 IFA 94 – DSM 19455added to silage is not achievable by analysis. Therefore the EURL cannot evaluate nor recommend any method for official control to determine Lactobacillus kefiri BIO 94 IFA 94 – DSM 19455 in silage. 

Molecular methods were used by the Applicant to identify the active agent in the feed additive. The EURL recommends 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.

¹New name: Lentilactobacillus buchneri DSM 19455.

Recommended Method

as of:

- Commission Implementing Regulation (EU) 2024/1196

Enumeration in the feed additive of Lentilactobacillus buchneri DSM 19455:

• Spread plate method using MRS Agar (EN 15787)

Identification of Lentilactobacillus buchneri DSM 19455:

• Pulsed Field Gel Electrophoresis (PFGE) – CEN/TS 17697

  or 

• DNA sequencing methods