Assessment of the feed additive consisting of Lentilactobacillus diolivorans DSM 32074 (1k20752) for all animal species for the renewal of its authorisation (Lactosan GmbH & Co.KG)

Abstract

Following a request from the European Commission, EFSA was asked to deliver a scientific opinion on the assessment of the application for renewal of authorisation of Lentilactobacillus diolivorans DSM 32074 as a technological additive for all animal species. The applicant provided evidence that the additive currently on the market complies with the existing terms of the authorisation. The Panel concluded that the additive remains safe for all animal species, consumers and the environment. Regarding user safety, the additive is a preparation containing a microorganism and therefore should be considered as a skin and respiratory sensitiser. Any exposure through skin and respiratory tract is considered a risk. The additive is not an eye irritant. There is no need for assessing the efficacy of the additive in the context of the renewal of the authorisation.

1. INTRODUCTION

1.1. Background and Terms of Reference

Regulation (EC) No 1831/2003 ¹ establishes the rules governing the Community authorisation of additives for use in animal nutrition. In particular, Article 14(1) of that Regulation lays down that an application for renewal shall be sent to the Commission at the latest one year before the expiry date of the authorisation.

The European Commission received a request from Lactosan GmbH & Co.KG ² for the renewal of the authorisation of the additive consisting of Lentilactobacillus diolivorans DSM 32074, when used as a feed additive for all animal species (category: technological additives; functional group: silage additives).

According to Article 7(1) of Regulation (EC) No 1831/2003, the Commission forwarded the application to the European Food Safety Authority (EFSA) as an application under Article 14(1) (renewal of the authorisation). The dossier was received on 14 January 2026 and the general information and supporting documentation are available at https://open.efsa.europa.eu/questions/EFSA‐Q‐2026‐00028. The particulars and documents in support of the application were considered valid by EFSA as of 10 April 2026.

According to Article 8 of Regulation (EC) No 1831/2003, EFSA, after verifying the particulars and documents submitted by the applicant, shall undertake an assessment in order to determine whether the feed additive complies with the conditions laid down in Article 5. EFSA shall deliver an opinion on the safety for the target animals, consumer, user and the environment and on the efficacy of the feed additive consisting of L. diolivorans DSM 32074, when used under the proposed conditions of use (see Section 3.1.3 ).

1.2. Additional information

The additive L. diolivorans (formerly Lactobacillus diolivorans) DSM 32074 is currently authorised for use in feed for all animal species (1k20752). ³

EFSA issued an opinion on the safety and efficacy of this product when used in feed for all animal species (EFSA FEEDAP Panel, 2016).

2. DATA AND METHODOLOGIES

2.1. Data

The present assessment is based on data submitted by the applicant in the form of a technical dossier ⁴ in support of the authorisation request for the use of L. diolivorans DSM 32074 as a feed additive.

In accordance with Article 38 of the Regulation (EC) No 178/2002 ⁵ and taking into account the protection of confidential information and of personal data in accordance with Articles 39 to 39e of the same Regulation, and of the Decision of EFSA's Executive Director laying down practical arrangements concerning transparency and confidentiality, ⁶ a non‐confidential version of the dossier has been published on Open.EFSA.

According to Article 32c(2) of Regulation (EC) No 178/2002 and to the Decision of EFSA's Executive Director laying down the practical arrangements on pre‐submission phase and public consultations, EFSA carried out a public consultation on the non‐confidential version of the technical dossier from 30 June 2026 to 21 July 2026 for which no comments were received.

The confidential version of the technical dossier was subject to a target consultation of the interested Member States from 15 April 2026 to 15 July 2026. No comments were received.

The European Union Reference Laboratory (EURL) considered that the conclusions and recommendations reached in the previous assessment regarding the methods used for the control of the active substance in animal feed are valid and applicable for the current application. ⁷

2.2. Methodologies

The approach followed by the FEEDAP Panel to assess the safety and the efficacy of L. diolivorans DSM 32074 is in line with the principles laid down in Regulation (EC) No 429/2008 ⁸ and the relevant guidance documents: Guidance on the renewal of the authorisation of feed additives (EFSA FEEDAP Panel, 2021), EFSA statement on the requirements for whole genome sequence analysis of microorganisms intentionally used in the food chain (EFSA, 2024).

3. ASSESSMENT

The additive consisting of L. diolivorans DSM 32074 is currently authorised as a technological additive (functional group: silage additives) for all animal species. The applicant is requesting the renewal of the above authorisation.

3.1. Characterisation

3.1.1. Characterisation of the additive

The additive is currently authorised with a minimum content of L. diolivorans DSM 32074 of 3 × 10¹¹ colony forming units (CFU)/g additive. The applicant declared minor changes in the manufacturing process since the previous authorisation, removing PEG 4000 as a cryoprotectant and including it as a possible carrier. No antimicrobials are used during the manufacturing process. ⁹

The final additive consists of approximately 35%–50% active agent and 50%–65% cryoprotectants/carriers. ¹⁰ All the proposed cryoprotectants and carriers are feed materials and/or authorised feed additives, ¹¹ with the exception of PEG 4000 (E1521), the safety of which is assessed in Section 3.2 .

The data provided by the applicant on the batch‐to‐batch variation, ¹² impurities ¹³ and dusting potential ¹⁴ of the additive are reported in Table 1 .

TABLE 1.

Data on the batch‐to‐batch variation, impurities and physico‐chemical and technological properties of the additive. The data presented are average values and (range) for batch‐to‐batch variation, and ranges for all other parameters. The number of batches analysed per parameter or group of parameters is indicated in [].

Specifications 1
L. diolivorans DSM 32074 (CFU/g)	≥ 3 × 1011
Batch‐to‐batch variation	[5]
L. diolivorans DSM 32074 (CFU/g) 2	3.83 × 1011 (3.66 – 4.00 × 1011)
Impurities	[3]
Lead (mg/kg)	< 0.10
Mercury (mg/kg)	< 0.10
Cadmium (mg/kg)	< 0.10
Arsenic (mg/kg)	< 0.10
Mycotoxins (μg/kg)	[3]
Aflatoxins B1, B2, G1, G2 (individually)	< 0.03
Zearalenone	< 10
Deoxynivalenol	< 20
Microbial contamination	[3]
Salmonella spp. (per 25 g)	Not detected
Enterobacteriaceae (CFU/g)	< 103
Yeast and moulds (CFU/g)	< 103
Physical properties
Physical form	Solid
Dusting potential (Stauber Heubach) (mg/m3)	814–859 [3]

Note: <: below the limit of quantification.

¹ Specifications set in Regulation (EU) No 194/2017.

² Identification confirmed by molecular method.

The data provided showed compliance with the specifications set in the authorising regulation for L. diolivorans DSM 32074. ¹⁵ The Panel considers that the microbial contamination and the levels of the impurities analysed are of no concern. No new data were provided regarding the physico‐chemical and technological properties of the additive, with the exception of the dusting potential. Since no significant changes were introduced in the manufacturing process and composition, the data described in the previous opinions (EFSA FEEDAP Panel, 2016) are considered still valid.

3.1.2. Characterisation of the active agent

The active agent was originally isolated from silage, and it is deposited at the German collection of microorganisms and cell cultures (DSMZ) with accession number DSM 32074. ¹⁶ The applicant declared that it has not been genetically modified. ¹⁷

The taxonomical identification of the active agent was confirmed ■■■■■ based on the whole genome sequence (WGS). ¹⁸ ■■■■■

The antimicrobial susceptibility of the strain L. diolivorans DSM 32074 was tested using a broth microdilution method against the battery of antibiotics recommended by the EFSA FEEDAP Panel (EFSA FEEDAP Panel, 2018a). ¹⁹ All the minimum inhibitory concentration (MIC) values were equal to or fell below the corresponding cut‐off values for ‘Lactobacillus obligate heterofermentative’ (EFSA FEEDAP Panel, 2018a), except for tetracycline (MIC: 16 mg/L vs. cut‐off value: 8 mg/L). Exceeding the cut‐off value by one dilution is considered to be within the normal range of variation and, thus, not a matter of concern. Therefore, the strain is considered to be susceptible to all the relevant antibiotics.

The WGS data of the strain DSM 32074, including the plasmids, were interrogated for the presence of antimicrobial resistance (AMR) genes by a search against ResFinder and AMRFinderPlus databases, ²⁰ using equal or below thresholds as indicated by the EFSA statement (EFSA, 2024). The search did not identify any hit above the thresholds established by the EFSA statement, and therefore, the strain does not harbour acquired AMR genes.

3.1.3. Conditions of use

The additive is currently authorised as a silage additive for all animal species. Under other provisions of the authorisation, ²¹ it is specified that:

• In the directions for use of the additive and premixture, indicate the storage conditions.

• Minimum content of the additive when used without combination with other micro‐organisms as silage additives: 1 × 10⁸ CFU/kg fresh material.

• For users of the additive and premixtures, feed business operators shall establish operational procedures and organisational measures to address potential risks resulting from its use. Where those risks cannot be eliminated or reduced to a minimum by such procedures and measures, the additive and premixtures shall be used with personal protective equipment, including breathing protection.

The applicant did not request any change in the current conditions of the authorisation.

3.2. Safety

In its previous opinion, the FEEDAP Panel concluded that, following the qualified presumption of safety (QPS) approach to the safety assessment, the use of the strain L. diolivorans DSM 32074 as a silage additive is safe for the target species, consumers and the environment (EFSA FEEDAP Panel, 2016). Regarding user safety, the Panel concluded that ‘The additive is not a skin or eye irritant but should be considered to have the potential to be a respiratory sensitiser. In the absence of data, no conclusion can be drawn on the dermal sensitisation potential of theadditive.’

The applicant declared that no incidents or safety issues for target animals, consumers, users and/or the environment have been documented or reported regarding the additive since its approval. ²²

In the context of the current application, the identity of the strain as L. diolivorans was confirmed, and evidence that the strain does not carry acquired genes coding for therapeutic antimicrobials was provided (EFSA BIOHAZ Panel, 2026). Consequently, the conclusions previously reached are still valid, and the Panel considers that L. diolivorans DSM 32074 remains safe for the target species, consumers and the environment.

An extensive literature search was performed to support the safety of the additive for the target species, consumers, users and the environment. The search covered the period from January 2000 to September 2025. The terms used included the active agent at strain level combined with a set of terms related to safety. The applicant searched a total of nine databases: Agris, Basenet, Europe PMC, Google scholar, Ingenta, National Agricultural Library, PubMed, Science Direct and World Cat. The literature search retrieved 54 hits. None of the articles described safety concerns related to the use of the additive. ²³

All the proposed cryoprotectants and carriers are feed materials or authorised feed additives, with the exception of PEG 4000 that is authorised as a food additive.

The additive may contain up to 9.1% of PEG 4000 according to the applicant declaration, which corresponds to a concentration of 0.025 mg PEG 4000/kg silage. ²⁴ The safety of PEG 4000 used at the same concentration in the formulation of another silage additive has already been assessed by the FEEDAP Panel in previous opinions (EFSA FEEDAP Panel, 2018b, 2024). The FEEDAP Panel concluded based on a worst‐case scenario that, at this concentration, PEG 4000 is safe for target animals, consumers, users and the environment. The same conclusion applies to the present assessment.

Considering all the above, the Panel concludes that the use of preparations formulated with the cryoprotectants and carriers listed by the applicant is safe for the target animals, consumers and the environment.

Considering that the composition of the additive and the manufacturing process have not been modified and that there is no new information that would lead the Panel to reconsider its previous assessment, the FEEDAP Panel considers that the previous conclusions still apply. Therefore, the FEEDAP Panel concludes that the additive remains safe for the target species, consumers and the environment. Regarding user safety, the additive consists of (or contains) a microorganism and therefore should be considered as a skin and respiratory sensitiser. Exposure of users via dermal and respiratory routes is considered a risk. These conclusions would apply, in principle, to any preparations containing the active agent.

3.3. Efficacy

The present application for renewal of the authorisation does not include a proposal for amending or supplementing the conditions of the original authorisation that would have an impact on the efficacy of the additive. Therefore, there is no need for assessing the efficacy of the additive in the context of the renewal of the authorisation.

4. CONCLUSIONS

The applicant provided evidence that the additive currently in the market complies with the existing conditions of the authorisation.

The FEEDAP Panel concludes that the preparation of L. diolivorans DSM 32074 remains safe for all animal species, consumers and the environment.

Regarding user safety, the additive should be considered as a skin and respiratory sensitiser. Exposure of users via dermal and respiratory routes is considered a risk. This conclusion would apply, in principle, to any preparations containing the active agent.

There is no need for assessing the efficacy of the additive in the context of the renewal of the authorisation.

ABBREVIATIONS

AMR

antimicrobial resistance

CFU

colony forming unit

DM

dry matter

EURL

European Union Reference Laboratory

FEEDAP

EFSA Scientific Panel on Additives and Products or Substances used in Animal Feed

LOD

limit of detection

LOQ

limit of quantification

MIC

minimum inhibitory concentration

QPS

qualified presumption of safety

WGS

whole genome sequence

AUTHOR CONTRIBUTIONS

Under Regulation 178/2002, the Scientific Panel is the author of the scientific output. Contributions include Working Groups who provide the draft assessment which is reviewed and adopted by the Panel; EFSA staff members who provide scientific support including inter alia data collection and analysis and literature searching; and other contributors such as staff of national food safety agencies, EFSA trainees and interim staff members who have contributed meaningfully to the work.

REQUESTOR

European Commission

QUESTION NUMBERS

EFSA‐Q‐2026‐00028

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LEGAL NOTICE

The published scientific output implements EFSA's decision on the confidentiality requests submitted on specific information items. Information items awarded confidential status by EFSA and included in the relevant scientific output have been withheld from public disclosure by redaction.

EFSA FEEDAP Panel (EFSA Panel on Additives and Products or Substances used in Animal Feed) , Villa, R. E. , Azimonti, G. , Bonos, E. , Christensen, H. , Durjava, M. , Dusemund, B. , Gehring, R. , Glandorf, B. , Kouba, M. , López‐Alonso, M. , Marcon, F. , Martelli, G. , Nebbia, C. , Pechová, A. , Prieto‐Maradona, M. , Theodoridou, K. , Anguita, M. , Innocenti, M. L. , … García‐Cazorla, Y. (2026). Assessment of the feed additive consisting of Lentilactobacillus diolivorans DSM 32074 (1k20752) for all animal species for the renewal of its authorisation (Lactosan GmbH & Co.KG) EFSA Journal, 24(10), e10352. 10.2903/j.efsa.2026.10352