Assessment of the feed additive consisting of Bacillus velezensis ATCC PTA‐6737 (PB6) for the renewal of its authorisation for laying hens and minor poultry species for laying (4b1823) and its extension of use to other laying/breeding birds (Kemin Europa n.v.)

TL;DR

This paper evaluates the safety and efficacy of a feed additive containing Bacillus velezensis for laying hens and other egg-producing birds.

Contribution

The study confirms the safety and efficacy of a higher concentration of the additive for poultry and proposes an extension of its use.

Findings

The additive is safe for laying hens, egg-producing birds, consumers, and the environment.

The additive is not irritant to skin and eyes but is a potential skin and respiratory sensitiser.

The additive is efficacious at a minimum inclusion level of 1 × 10⁸ CFU/kg complete feed.

Abstract

Following a request from the European Commission, EFSA was asked to deliver a scientific opinion on the safety and efficacy of a feed additive consisting of Bacillus velezensis ATCC PTA‐6737 (PB6) as a zootechnical feed additive. With the current application, the applicant is seeking the renewal of the authorisation for laying hens and minor poultry species for laying. In addition, the applicant is seeking the modification of the terms of the existing authorisation to obtain the authorisation of a more concentrated form of the additive (from 1 × 1010 to 8 × 1010 colony forming units (CFU)) and the authorisation of the additive for other birds kept for egg production or breeding. The evidence provided showed that the additive complies with the new specifications and the current terms of the authorisation. The FEEDAP Panel concludes that the additive is safe for laying hens, birds kept for egg production or breeding, the consumer and the environment. Regarding safety for the user, the additive is not irritant to the skin and eyes but should be considered a skin and respiratory sensitiser and inhalation and dermal exposure is considered a risk. The additive has the potential to be efficacious in other birds kept for egg production or breeding at the minimum inclusion level of 1 × 108 CFU/kg complete feed.

Tables

TABLE 1. Data on the batch‐to‐batch variation and impurities of PB6.

		Analysis
Parameter	Specification	Average	Range	# batches
Batch to batch variation
Bacillus velezensis ATCC PTA‐6737 (CFU/g)	1 × 1010 (1) 8 × 1010 (2)	1.06 × 1011	1.0–1.1 × 1011	5
Impurities
Lead (mg/kg)			< 0.20–1.3	5
Mercury (mg/kg)			< 0.01–0.011	5
Cadmium (mg/kg)			< 0.05–0.43	5
Arsenic (mg/kg)			< 0.20–0.48	5
Fluoride (mg/kg)			49.8–152	3
Mycotoxins (μg/mg)
Aflatoxins B1			< 0.05–1	5
Aflatoxins G1			< 1	5
Aflatoxins B2			< 1	5
Aflatoxins G2			< 1	5
Acetyldeoxynivalenol			< 20	5
Fumonisins B1 + B2			< 20	5
Zearalenone			< 15	5
Nivalenol			< 50	5
Cytochalasine E			< 2	5
Ochratoxin A			< 1	5
Deoxynivalenol			< 20	5
T2‐toxin			< 10	5
HT2‐toxin			< 10	5
Dioxin + Dioxin‐like PCBs (ng WHO2005‐TEQ/kg)			0.13–0.14	3
Microbial contamination
Salmonella spp. (per 25 g)			Not detected	10
Enterobacteriaceae (CFU/g)			< 10	2
Yeasts and moulds (CFU/g)			< 10	10
Coliforms (per 10 g)			Not detected	8
Escherichia coli (per 10 g)			Not detected	8
Staphylococcus aureus (per 10 g)			Not detected	8
Bacillus cereus (CFU/g)			< 100	2

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. In particular, Article 4(1) of that Regulation lays down that any person seeking authorisation for a feed additive or for a new use of feed additive shall submit an application in accordance with Article 7.

The European Commission received a request from Kemin Europa n.v.2 for the renewal of the authorisation of the additive consisting of Bacillus velezensis ATCC PTA‐6737 (PB6) when used in laying hens and minor poultry species for laying and the authorisation of a new use in other birds kept for egg production or breeding (category: zootechnical additives; functional group: gut flora stabilisers).

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 4(1) (authorisation of a feed additive or new use of a feed additive) and under Article 14(1) (renewal of the authorisation). The dossier was received on 12 June 2024 and the general information and supporting documentation are available at https://open.efsa.europa.eu/questions/EFSA‐Q‐2024‐00395. The particulars and documents in support of the application were considered valid by EFSA as of 8 October 2024.

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 Bacillus velezensis ATCC PTA‐6737 (PB6), when used under the proposed conditions of use (see Section 3.1.3).

Additional information

1.2

The additive under assessment is a preparation consisting of viable spores of a strain of Bacillus velezensis ATCC PTA‐6737. It is currently authorised for use in feed for chickens for fattening, chickens reared for laying, minor poultry species except minor poultry for laying, turkeys for fattening, turkeys reared for breeding, laying hens, minor poultry species for laying, ornamental birds and for piglets, pigs for fattening and sows of all Suidae species (4b1823 and 4b1823i).3 ^,^ 4

EFSA has issued several opinions on the safety and efficacy of Bacillus subtilis 5 ATCC PTA‐ 6737 (Bacillus subtilis PB6) (EFSA FEEDAP Panel, 2009, 2010, 2011, 2012, 2013, 2015, 2017a, 2020, 2022, 2024a, 2024b).

DATA AND METHODOLOGIES

2

Data

2.1

The present assessment is based on data submitted by the applicant in the form of a technical dossier6 in support of the authorisation request for the use of B. velezensis ATCC PTA‐6737 (PB6) as a feed additive.

The confidential version of the technical dossier was subject to a target consultation of the interested Member States from 14 October 2024 to 14 January 2025; the comments received were considered for the assessment.

In accordance with Article 38 of the Regulation (EC) No 178/20027 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,8 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 27 March to 17 April 2025 for which no comments were received.

The FEEDAP Panel used the data provided by the applicant together with data from other sources, such as previous risk assessments by EFSA to deliver the present output.

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 B. velezensis ATCC PTA‐6737 in animal feed are valid and applicable for the current application.9

Methodologies

2.2

The approach followed by the FEEDAP Panel to assess the safety and the efficacy of B. velezensis ATCC PTA‐6737 (PB6) is in line with the principles laid down in Regulation (EC) No 429/200810 and the relevant guidance documents: Guidance on the assessment of the safety of feed additives for the consumer (EFSA FEEDAP Panel, 2017b), Guidance on the identity, characterisation and conditions of use of feed additives (EFSA FEEDAP Panel, 2017c), Guidance on the assessment of the safety of feed additives for the target species (EFSA FEEDAP Panel, 2017d), Guidance on the characterisation of microorganisms used as feed additives or as production organisms (EFSA FEEDAP Panel, 2018), Guidance on the assessment of the safety of feed additives for the environment (EFSA FEEDAP Panel, 2019), Guidance on the renewal of the authorisation of feed additives (EFSA FEEDAP Panel, 2021), Guidance on the assessment of the safety of feed additives for the users (EFSA FEEDAP Panel, 2023), Guidance on the assessment of the efficacy of feed additives (EFSA FEEDAP Panel, 2024c) and EFSA statement on the requirements for whole genome sequence analysis of microorganisms intentionally used in the food chain (EFSA, 2024).

ASSESSMENT

3

The preparation consisting of viable spores of B. velezensis ATCC PTA‐6737 is currently authorised for use as a zootechnical additive (functional group: gut flora stabilisers) in feed for chickens for fattening, chickens reared for laying, turkeys for fattening and reared for breeding, minor poultry species except minor poultry for laying, laying hens, minor poultry species for laying, ornamental birds, piglets, pigs for fattening and sows of all Suidae species. The applicant is requesting the renewal of the authorisation for laying hens and minor poultry species for laying, the modification of the terms of the authorisation as regards the concentration of the active agent in the additive, and the authorisation of the additive for other birds kept for egg production or breeding. The additive will be hereafter referred to as its tradename PB6.

Characterisation

3.1

Characterisation of the active agent

3.1.1

The active agent B. velezensis is a non‐genetically modified microorganism originally isolated from the intestinal tract of a chicken and deposited in the American Type Culture Collection with the accession number ATCC PTA‐6737. It was fully characterised in the previous opinions (EFSA FEEDAP Panel, 2024a, 2024b). In those instances, the taxonomic identification of the active agent was confirmed based on a bioinformatic analysis, the strain proved to be susceptible to all the relevant antibiotics and not to have toxigenic potential. These conclusions are still considered valid. In the present application, some new data have been submitted that are described below.

The WGS data of the active agent were interrogated for the presence of antimicrobial resistance (AMR) genes by a search against the ResFinder and CARD databases and three hits exceeding the established thresholds were identified (EFSA, 2024).11 Further analyses of these hits following EFSA's criteria (EFSA BIOHAZ Panel, 2023) did not reveal any acquired AMR gene. Therefore, the FEEDAP Panel concludes that the strain does not harbour acquired AMR genes and raises no safety concerns.

Characterisation of the additive

3.1.2

The additive currently authorised is a preparation containing viable spores of B. velezensis ATCC PTA‐6737 at the minimum concentration of 1 × 10^10^ colony forming units (CFU) per gram of additive. In the current application, the applicant proposes a modification of the specifications of the additive, in particular an increase of the minimum concentration from 1 × 10^10^ CFU/g to 8 × 10^10^ CFU/g additive.

The additive in its current form has the same method of manufacture as that considered in the previous opinions (EFSA FEEDAP Panel, 2024a, 2024b), and essentially the same composition (i.e. spores concentrate (~5%–25%) and sodium bicarbonate (75%–95%)) except for the concentration of the active agent which has been increased. The Panel considers that the data pertaining to physico‐chemical properties, shelf‐life, stability and homogeneity described in those opinions apply to the current assessment.

New data have been provided to support the compliance with the new specifications of the additive12 and on purity13 which are described below (Table 1).

The data provided showed compliance with the newly proposed specifications. The FEEDAP Panel considers that the amounts of the detected impurities do not raise safety concerns.

Conditions of use

3.1.3

The additive is currently authorised for use in laying hens and minor poultry species for laying at the minimum inclusion level of 1 × 10^8^ CFU/kg complete feed.

Under other provisions of the authorisation, it is specified that:

• –In the directions for use of the additive and premixture indicate the storage conditions and stability to pelleting.

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

The applicant is requesting the extension of use of the additive in feed for other birds kept for egg production or breeding at the same minimum inclusion level of 1 × 10^8^ CFU/kg complete feed.

Safety

3.2

For the present application, the applicant states that no adverse events have been reported since the market authorisation of the additive (2015).14

In the previous assessments (EFSA FEEDAP Panel, 2024a, 2024b), B. velezensis ATCC PTA‐6737 was considered to meet the requirements of the qualified presumption of safety (QPS) approach and was presumed safe for the target animals, consumers and the environment. Moreover, since no safety concerns are expected from other components of the additive, the additive was also considered to be safe for the target species, consumers and the environment. The data provided in the present application demonstrated the absence of acquired AMR genes, thus confirming the applicability of the QPS approach. Therefore, the FEEDAP Panel concludes that the use of the additive under the approved conditions remains safe for laying hens and minor poultry species for laying, the consumers and the environment. As the exposure of animals will be the same, the FEEDAP Panel considers that the use of a more concentrated form of the additive will not introduce new safety concerns not already considered in the previous assessments for the target species, the consumers and the environment.

The FEEDAP Panel considers that the extension of use of the additive in feed for other birds kept for egg production or breeding would not add concerns to those already identified in the previous opinions. Therefore, the Panel concludes that this use is also safe for the new target species, the consumers and the environment.

In a previous opinion (EFSA FEEDAP Panel, 2022), the FEEDAP Panel concluded that the additive was non‐irritant to skin and eyes or a dermal sensitiser but should be considered a respiratory sensitiser. No additional data were submitted in the current application. The additive under assessment consists of microorganisms and therefore, it should be considered a skin and respiratory sensitiser and inhalation, and dermal exposure is considered a risk.

Conclusions on safety

3.2.1

Considering the above and the fact that the manufacturing process of the additive have not been modified, the FEEDAP Panel concludes that the additive remains safe for laying hens, minor poultry species for laying, consumers, and the environment. The modifications in the specifications of the additive do not introduce additional concerns. The FEEDAP Panel concludes that the additive is safe for birds kept for egg production or breeding, and that the use of the additive in these new species/categories is safe for the consumer and the environment.

The additive is not irritant to the skin and eyes but should be considered a skin and respiratory sensitiser and inhalation and dermal exposure is considered a risk.

Efficacy

3.3

The present application for renewal of the authorisation does not include a proposal for amending or supplementing the conditions of use for the species/categories for which an authorisation exists. Therefore, there is no need for assessing the efficacy of the additive in the context of the renewal of the authorisation.

The applicant requested the extension of use of the additive to birds kept for egg production or breeding at a minimum inclusion level of 1 × 10^8^ CFU/kg of complete feed.

Considering that the efficacy of the additive was demonstrated in laying hens (EFSA FEEDAP, 2015), the FEEDAP Panel, in line with the provisions of the guidance on the assessment of the efficacy of feed additives (EFSA FEEDAP Panel, 2024a, 2024b), concludes that PB6 has the potential to be efficacious in birds kept for egg production or breeding at the minimum inclusion level of 1 × 10^8^ CFU/kg complete feed.

Post‐market monitoring

3.4

The FEEDAP Panel considers that there is no need for specific requirements for a post‐market monitoring plan other than those established in the Feed Hygiene Regulation15 and Good Manufacturing Practice.

CONCLUSIONS

4

The applicant has proposed a modification of the specifications of the additive PB6 by increasing the minimum concentration of the active agent from 1 × 10^10^ to 8 × 10^10^ CFU/g additive. The evidence provided showed that the additive complies with the new specifications and the current terms of the authorisation.

PB6 remains safe for laying hens and minor poultry species for laying, the consumers, and the environment. The FEEDAP Panel concludes that the use of the additive in other birds kept for egg production or breeding is safe for the target species, the consumer and the environment.

Regarding safety for the user, PB6 is not irritant to the skin and eyes but should be considered a skin and respiratory sensitiser and inhalation and dermal exposure is considered a risk.

The FEEDAP Panel concludes that the additive has the potential to be efficacious in other birds kept for egg production or breeding at the minimum inclusion level of 1 × 10^8^ CFU/kg complete feed.ABBREVIATIONSAMRantimicrobial resistanceCFUcolony forming unitFEEDAPEFSA Scientific Panel on Additives and Products or Substances used in Animal FeedPCBpolychlorinated biphenylQPSqualified presumption of safetyTEQtoxic equivalent factorsWHOWorld Health Organization

REQUESTOR

European Commission

QUESTION NUMBER

EFSA‐Q‐2024‐00395

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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.