Assessment of the feed additive consisting of Lactiplantibacillus plantarum (formerly Lactobacillus plantarum) NCIMB 30083 for all animal species for the renewal of its authorisation (Chr. Hansen A/S)

Abstract

Following a request from the European Commission, the Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) was asked to deliver a scientific opinion on the assessment of the application for renewal of authorisation of Lactiplantibacillus plantarum (previously Lactobacillus plantarum) NCIMB 30083 as a technological additive, silage additive for all animal species. The applicant has provided evidence that the additive currently on the market complies with the existing conditions of authorisation. There is no evidence that would lead the FEEDAP Panel to reconsider its previous conclusions. Thus, the Panel concluded that the additive remains safe for all animal species, consumers and the environment under the authorised conditions of use. Regarding user safety, the additive should be considered as a respiratory sensitiser. No conclusions can be drawn on the skin sensitisation, and skin and eye irritancy potential of the additive. 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 1 year before the expiry date of the authorisation.

The European Commission received a request from Chr. Hansen A/S ² for the renewal of the authorisation of the additive consisting of Lactiplantibacillus plantarum (formerly Lactobacillus plantarum) NCIMB 30083, when used as a feed additive for all animal species (category: technological; functional group: silage).

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 19 May 2022 and the general information and supporting documentation are available at https://open.efsa.europa.eu/questions/EFSA‐Q‐2022‐00317. The particulars and documents in support of the application were considered valid by EFSA as of 5 December 2022.

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 Lactiplantibacillus plantarum NCIMB 30083, when used under the proposed conditions of use (see Section 3.1.3 ).

1.2. Additional information

The additive is a preparation containing Lactiplantibacillus plantarum NCIMB 30083 is currently authorised for use in feed for all animal species (1 k20736). ³

EFSA issued an opinion on the safety and efficacy of this product when used in forages for all animal species (EFSA FEEDAP Panel, 2013).

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 Lactiplantibacillus plantarum (previously Lactobacillus plantarum) NCIMB 30083 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, ⁶ EFSA carried out a public consultation on the non‐confidential version of the technical dossier from 23 May to 13 June 2023 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 5 December 2022 to 6 March 2023 for which the received comments were considered for the assessment.

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 agent 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 Lactiplantibacillus plantarum NCIMB 30083 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).

3. Assessment

The product consisting of viable cells of Lactiplantibacillus plantarum NCIMB 30083 (previously Lactobacillus plantarum NCIMB 30083) is currently authorised as a technological additive (functional group: silage additive) for use in silage material for all animal species. This assessment regards the renewal of the authorisation of L. plantarum NCIMB 30083 as a silage additive.

3.1. Characterisation

3.1.1. Characterisation of the additive

The additive is currently authorised with a minimum content of the active agent (Lactiplantibacillus plantarum NCIMB 30083) of 5 × 10¹⁰ colony forming units (CFU)/g of additive. The applicant declared that the manufacturing process and the composition have not been changed since the previous authorisation and that no antimicrobials are used during the manufacturing process. ⁹ The active agent is grown by fermentation and concentrated by centrifugation and filtration. Cryoprotectant agents (17 –42%) are added and the cell concentrate is freeze‐dried. The final product is a powder containing the freeze‐dried cell concentrate (which include a maximum of 6% of fermentation medium, a maximum of 3% of water and 17–42% of cryoprotectants), synthetic amorphous silica as anticaking agent (8%) ¹⁰ and maltodextrin as carrier (50–75%), to reach the minimum concentration specified for L. plantarum NCIMB 30083. ¹¹

Analytical data to confirm the specification were provided for nine samples of the additive, showing an average value of 6.6 × 10¹¹ CFU/g additive (range from 6.2 × 10¹¹ to 7.2 × 10¹¹ CFU/g additive ¹² ). ¹³ However, the Panel notes that these samples were obtained from only four independent fermentation batches. ¹⁴

Six recent batches (three independent) of the additive were tested for arsenic (range 0.017–0.051 mg/kg), lead (< 0.01 mg/kg), cadmium (range 0.015–0.034 mg/kg) and mercury (range 0.0035–0.0089 mg/kg). Aflatoxin B1 was tested in the same batches resulting < 0.46 μg/kg. ¹⁵

Specifications are set for coliforms (< 1,000 CFU/g), Escherichia coli (< 10 CFU/g), Salmonella spp. (no detection in 25 g frozen product or 5 g freeze dried bulk) and yeasts and filamentous fungi (< 1,000 CFU/g). Nine samples of the additive from four independent fermentation batches showed compliance with the respective specifications. The Panel notes that the specification for Salmonella spp. detection has been modified and that data from analyses of 5 g instead of 25 g were provided for four out of five batches. In addition, the applicant provided data on Enterobacteriaceae for three batches, with results below the limit of quantification (< 10 CFU/g). 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. Since no changes were introduced in the manufacturing process, the data described in the previous opinion (EFSA FEEDAP Panel, 2013) still apply.

3.1.2. Characterisation of the active agent

Lactiplantibacillus plantarum NCIMB 30083 was isolated from silage and it is deposited in The National Collections of Industrial and Marine Bacteria (NCIMB) with the accession number NCIMB 30083. ¹⁷ It has not been genetically modified.

The taxonomical identification of L. plantarum NCIMB 30083 was confirmed by average nucleotide identity (ANI) determination using whole genome sequence (WGS) data. The ANI value obtained was 99.9% with the type strain Lactiplantibacillus plantarum DSM 20174^T. ¹⁸

The susceptibility of the strain L. plantarum NCIMB 30083 to antimicrobials was tested using a broth microdilution method and including the set of antibiotics recommended by EFSA (EFSA FEEDAP Panel, 2018). ¹⁹ All the minimum inhibitory concentration values were below the cut‐off values, except for kanamycin, which was one dilution higher. Exceeding the cut‐off by one dilution is considered within the normal range of variation, and therefore, the strain is considered susceptible to all the relevant antibiotics.

The WGS data of the strain were interrogated for the presence of antimicrobial resistance (AMR) genes against two databases. The interrogation was performed with a threshold of ■■■■■ identity and ■■■■■ coverage at protein level with the NCBI Bacterial Antimicrobial Resistance Reference Gene database and at nucleotide level with ResFinder. No hits of concern were identified. ²⁰

3.1.3. Conditions of use

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

• In the directions for the use of the additive, indicate the storage temperature and storage life.

• Minimum dose of the additive when it is not used in combination with other microorganisms as silage additive: 1 × 10⁸ CFU/kg of fresh material.

• The additive shall be used in easy and moderately difficult to ensile material.

• For safety: It is recommended to use breathing protection and gloves during handling.

The applicant has requested to maintain the same conditions of use.

3.2. Safety

The applicant stated that no adverse effects or incidents/accidents have been reported from the use of the feed additive since the first authorisation of the product. ²¹

3.2.1. Safety for the target species, consumers and environment

In the previous opinion, the Panel concluded that following the qualified presumption of safety (QPS) approach, the use of L. plantarum NCIMB 30083 in the production of silage was considered safe for target species, consumers and the environment (EFSA FEEDAP Panel, 2013). In the context of this application, the identity of the strain as L. plantarum was confirmed, and evidence that the strain does not show acquired antimicrobial determinants for antibiotics of human and veterinary importance was provided (EFSA, 2007; EFSA BIOHAZ Panel, 2023). Consequently, the conclusions already reached are still valid, and the Panel considers that L. plantarum NCIMB 30083 remains safe for the target species, consumers and the environment.

An extensive literature search, however, has been performed to support the safety of the L. plantarum species and the L. plantarum NCIMB 30083 strain. The literature search presented several methodological limitations and, therefore, was not further considered for the assessment. ²²

3.2.2. Safety for the user

In the previous assessment (EFSA FEEDAP Panel, 2013), the Panel concluded regarding user safety: ‘In the absence of data the potential of the additives to be irritants and/or to act as skin/respiratory sensitisers cannot be totally excluded. […] Given the proteinaceous nature of the active agent, the additive should be considered to have the potential to be a skin/respiratory sensitiser and treated accordingly’.

No specific data have been submitted for the renewal of the authorisation regarding the effects of the additive on the skin and eyes. Therefore, the FEEDAP Panel cannot conclude on the eye/skin irritancy potential of the additive. The FEEDAP Panel notes that the OECD test guidelines available at present are designed to assess the skin sensitisation potential of chemical substances only and that currently no validated assays for assessing the sensitisation potential of microorganisms are available. Therefore, no conclusion can be drawn on the skin sensitisation potential of the additive. Owing to the proteinaceous nature of the active agent, the additive should be considered a respiratory sensitiser.

3.2.2.1. Conclusions on safety

The FEEDAP Panel concludes that there is no new evidence that would lead to reconsider the previous conclusions that L. plantarum NCIMB 30083 is safe for the target species, consumers and the environment under the authorised conditions of use. No conclusions can be drawn on the skin sensitisation, and skin and eye irritancy potential of the additive. The additive should be considered as a respiratory sensitiser.

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

The Panel concludes that L. plantarum NCIMB 30083 remains safe for all animal species, consumers and the environment under the authorised conditions of use. Regarding user safety, the additive should be considered as a respiratory sensitiser. No conclusions can be drawn on the skin sensitisation, and skin and eye irritancy potential of the additive.

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

Abbreviations

CFU

colony forming unit

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

OECD

Organisation for Economic Co‐operation and Development

RH

relative humidity

Suggested citation: EFSA FEEDAP Panel (EFSA Panel on Additives and Products or Substances used in Animal Feed) , Bampidis, V ., Azimonti, G ., Bastos, M. L ., Christensen, H ., Dusemund, B ., Durjava, M ., Kouba, M ., López‐Alonso, M ., López Puente, S ., Marcon, F ., Mayo, B ., Pechová, A ., Petkova, M ., Ramos, F ., Sanz, Y ., Villa, R. E ., Woutersen, R ., Saarela, M ., … Amaduzzi, A ., (2023). Assessment of the feed additive consisting of Lactiplantibacillus plantarum (formerly Lactobacillus plantarum) NCIMB 30083 for all animal species for the renewal of its authorisation (Chr. Hansen A/S). EFSA Journal, 21(8), 1–8. 10.2903/j.efsa.2023.8154