Assessment of the feed additive consisting of Lactiplantibacillus plantarum CECT 4528 for all animal species for the renewal of its authorisation (Centro Sperimentale del Latte S.r.l.)

TL;DR

This paper evaluates the safety of a bacterial additive for animals and concludes it remains safe for use.

Contribution

The paper confirms the ongoing safety of Lactiplantibacillus plantarum CECT 4528 for all animal species and users.

Findings

The additive is safe for all animal species, consumers, and the environment.

It is a skin and respiratory sensitiser but not an eye irritant.

Efficacy assessment was not required for renewal.

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 Lactiplantibacillus plantarum CECT 4528 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.

Full text

INTRODUCTION

1

Background and Terms of Reference

1.1

Regulation (EC) No 1831/20031 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 Centro Sperimentale del Latte S.r.l.2 for the renewal of the authorisation of the additive consisting of Lactiplantibacillus plantarum CECT 4528, 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 26 April 2023 and the general information and supporting documentation are available at https://open.efsa.europa.eu/questions/EFSA‐Q‐2023‐00298. The particulars and documents in support of the application were considered valid by EFSA as of 26 September 2023.

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. plantarum CECT 4528, when used under the proposed conditions of use (see Section 3.1.3).

Additional information

1.2

The additive is a preparation containing L. plantarum (formerly Lactobacillus plantarum) CECT 4528. The additive is currently authorised for use in feed for all animal species (1k20746).3 EFSA has issued an opinion on the safety and efficacy of this product when used in feed for all animal species (EFSA FEEDAP Panel, 2014).

DATA AND METHODOLOGIES

2

Data

2.1

The present assessment is based on data submitted by the applicant in the form of a technical dossier4 in support of the authorisation request for the use of L. plantarum CECT 4528 as a feed additive.

In accordance with Article 38 of the Regulation (EC) No 178/20025 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 26 June to 17 July 2024 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 25 September 2023 to 25 December 2023 for which 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 L. plantarum CECT 4528 in animal feed are valid and applicable for the current application.7

Methodologies

2.2

The approach followed by the FEEDAP Panel to assess the safety and the efficacy of L. plantarum CECT 4528 is in line with the principles laid down in Regulation (EC) No 429/20088 and the relevant guidance documents: Guidance on the renewal of the authorisation of feed additives (EFSA FEEDAP Panel, 2021).

ASSESSMENT

3

The additive L. plantarum CECT 4528 is currently authorised as a technological additive (functional group: silage additives) for use in fresh material for all animal species. The assessment regards the renewal of the authorisation.

Characterisation

3.1

Characterisation of the additive

3.1.1

The additive is currently authorised as a preparation of L. plantarum CECT 4528 with a minimum content of the active agent of 2.5 × 10^11^ colony forming units (CFU)/g of additive. The applicant declared that the manufacturing process has not been modified since the previous assessment and that no antimicrobials are used during the manufacturing process.9

The additive, a powder containing the active agent and maltodextrin as cryoprotectant, is used to formulate preparations containing approximately 25%–35% cells, 25%–35% cryoprotectant and 30%–50% carriers (maltodextrin and/or lactose).10 The applicant provided data on the characterisation of the additive as a preparation formulated with maltodextrin.11

Five batches of the additive confirmed compliance with specifications of the additive as per authorisation, showing an average value of 3.4 × 10^11^ CFU/g additive (3.0–3.7 × 10^11^ CFU/g).12

The analysis of six batches of the additive13 showed compliance with the specifications set by the applicant for Enterobacteriaceae (< 100 CFU/g), yeasts and filamentous fungi (< 100 CFU/g) and Salmonella spp. (no detection in 25 g).14

Three batches of the additive were tested for arsenic, mercury, lead and cadmium and aflatoxin M1. All the samples showed levels below the limit of quantification (LOQ) of the analytical method for aflatoxin M1 and mercury.15 The levels of arsenic, cadmium and lead ranged 0.023–0.038, 0.022–0.024 and 0.010–0.024 mg/kg, respectively.16

The FEEDAP Panel considers that the microbial contamination and the amounts of the detected impurities do not raise safety concerns.

No new data were provided regarding the physico‐chemical properties or stability of the additive. Considering that no changes have been introduced in the manufacturing process and composition, the data described in the previous opinion (EFSA FEEDAP Panel, 2014) still apply.

Characterisation of the active agent

3.1.2

The active agent strain was isolated from ensiled grass. It is deposited in the Colección Española de Cultivos Tipo (CECT) under the accession number CECT 4528.17 The applicant claimed that it has not been genetically modified.18

The taxonomic identification of the strain was done by digital DNA–DNA hybridisation (dDDH) based on the whole genome sequence (WGS) data.19 Results showed the highest similarity of L. plantarum CECT 4528 with the type strain L. plantarum DSM 20174^T^ with a value of 89.4%. No plasmids were detected.

The antimicrobial susceptibility of the strain CECT 4528 was tested against the battery of antibiotics recommended by the FEEDAP Panel (EFSA FEEDAP Panel, 2018).20 All the minimum inhibitory concentration (MIC) values were equal to or fell below the cut‐off values established in the FEEDAP Guidance, except for chloramphenicol, gentamicin and kanamycin. The MIC value for chloramphenicol was one dilution above the cut‐off value (16 mg/L vs. 8 mg/L). Exceeding the cut‐off value by one dilution is considered to be within the normal range of variation of the method, and thus, not a matter of concern. The MIC values for gentamicin and kanamycin were two dilutions above the corresponding cut‐off values (64 mg/L vs. 16 mg/L and 256 mg/L vs. 64 mg/L, respectively). Therefore, the strain is susceptible to the relevant antibiotics, except for gentamicin and kanamycin.

The WGS of the strain was interrogated for the presence of antimicrobial resistance (AMR) genes by search against the ResFinder and NCBI databases.21 No hits were identified exceeding the thresholds recommended by EFSA (EFSA, 2021). Although the strain is resistant to gentamicin and kanamycin, since no acquired AMR genes were found in the WGS, these resistances do not raise safety concerns.

Conditions of use

3.1.3

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

• In the directions for use of the additive and premixture, indicate the storage temperature and storage life.
• Minimum content of the additive when used without combination with other microorganisms as silage additives: 1 × 10^9^ CFU/kg fresh material.
• For safety: It is recommended to use breathing protection, eye protection and gloves during handling.

The applicant did not request any change in the current conditions of the authorisation.22

Safety

3.2

In the previous assessment (EFSA FEEDAP Panel, 2014), the Panel concluded that, following the qualified presumption of safety (QPS) approach to safety assessment, the use of L. plantarum CECT 4528 in the production of silage was considered safe for target species, consumers and the environment. Regarding user safety, in the absence of data, the Panel concluded that the additive may cause irritation to skin/eyes and should be considered as a skin and respiratory sensitiser.

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.23

In the context of the current application, the identity of the strain as L. plantarum was confirmed, and evidence was provided that the strain does not harbour acquired antimicrobial resistance genes for antibiotics of human and veterinary importance, confirming the suitability of the QPS approach for safety assessment (EFSA BIOHAZ Panel, 2023). Consequently, the conclusions previously reached are still valid, and the panel considers that L. plantarum CECT 4528 remains safe for the target species, consumers and the environment.

A literature search was performed to support the safety of the L. plantarum CECT 4528 strain. The search was performed covering the time period from 2014 to 2024 and the terms used included the active agent and the keyword toxicity. The applicant searched a total of two databases: Web of Science and PubMed. The literature search did not retrieve any relevant hit.24

Regarding the safety for the user, the applicant tested the eye irritation potential of the additive according to OECD TG 492. The results showed that the additive is not an eye irritant (UN‐GHS ‘No Category’).25 The additive contains a microorganism and therefore should be considered as a skin and respiratory sensitiser.

Conclusions on safety

3.2.1

The FEEDAP Panel concludes that there is no new evidence that would lead it to reconsider the previous conclusions that L. plantarum CECT 4528 is safe for the target species, consumers and the environment. Regarding user safety, the additive is not irritant to the eyes but should be considered as a skin and respiratory sensitiser. Any exposure through skin and respiratory tract is considered a risk. These conclusions would apply, in principle, to any preparation containing the active agent.

Efficacy

3.3

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.

CONCLUSIONS

4

The applicant has provided evidence that the additive currently on the market complies with the existing terms of authorisation.

The Panel concludes that the additive L. plantarum CECT 4528 remains safe for all animal species, consumers and the environment.

Regarding user safety, the additive is not irritant to the eyes but should be considered as a skin and respiratory sensitiser, and any exposure through skin and respiratory tract is considered a risk.

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

ABBREVIATIONSAMRantimicrobial resistanceCFUcolony forming unitDMdry matterEURLEuropean Union Reference LaboratoryFEEDAPEFSA Scientific Panel on Additives and Products or Substances used in Animal FeedLODlimit of detectionLOQlimit of quantificationMICminimum inhibitory concentrationQPSqualified presumption of safetyWGSwhole genome sequence

REQUESTOR

European Commission

QUESTION NUMBER

EFSA‐Q‐2023‐00298

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PANEL MEMBERS

Roberto Edoardo Villa, Giovanna Azimonti, Eleftherios Bonos, Henrik Christensen, Mojca Durjava, Birgit Dusemund, Ronette Gehring, Boet Glandorf, Maryline Kouba, Marta López‐Alonso, Francesca Marcon, Carlo Nebbia, Alena Pechová, Miguel Prieto‐Maradona, Ilen Röhe and Katerina Theodoridou.