Safety and efficacy of a feed additive consisting of endo‐1,4‐beta‐xylanase produced with Bacillus subtilis LMG S‐15136 (Belfeed B MP/ML) (4a1606i) for all poultry species, suckling and weaned piglets, pigs for fattening, lactating sows and all minor porcine species other than reproductive animals for the renewal of its authorisation and the extension of its use to all Suidae (Puratos NV)

TL;DR

This paper evaluates the safety and efficacy of a feed additive containing endo-1,4-beta-xylanase for various animal species, confirming its safety and effectiveness under approved conditions.

Contribution

The study confirms the renewal of authorization and extension of use of the feed additive to all porcine species.

Findings

The additive Belfeed B MP/ML is safe for target species, consumers, and the environment under approved conditions.

The additive is efficacious in all porcine species at 10 IU endo-1,4-beta-xylanase/kg feed.

High microbial contamination levels in Belfeed B MP require monitoring during production.

Abstract

Following a request from the European Commission, EFSA was asked to deliver a scientific opinion on the safety and efficacy of the feed additive consisting of endo‐1,4‐beta‐xylanase (produced with Bacillus subtilis LMG S‐15136) (Belfeed B MP/ML) as a zootechnical additive (functional group: digestibility enhancers) in feed for all poultry species and Suidae. The additive is currently authorised for all poultry species, suckling and weaned piglets, pigs for fattening, lactating sows and all minor porcine species other than reproductive animals. This scientific opinion concerns the request for the renewal of the authorisation of the additive for these species and the extension of use to all porcine species/categories. The applicant provided evidence that the additive currently in the market complies with the conditions of the authorisations. The FEEDAP Panel concluded that the additive Belfeed B MP/ML remains safe for the target species, the consumer and the environment under the approved conditions of the authorisations. The extension of use to all porcine species would not affect these conclusions; therefore, the additive is considered safe for the target species, the consumer and the environment. Regarding the safety for the user, the two formulations of the additive are considered skin and eye irritants, and respiratory sensitisers. Any exposure of the user is considered a risk. There is no need for assessing the efficacy of the additive in the context of the renewal of the authorisation. The FEEDAP Panel concluded that the additive is efficacious in all porcine species/categories at 10 IU endo‐1,4‐beta‐xylanase/kg complete feed. The Panel noted that the analytical values found for microbial contamination in Belfeed B MP are high and deserve attention/monitoring during the production process.

Tables

TABLE 1. Data on the batch‐to‐batch variation, impurities and physical properties of Belfeed. The data presented are average values, (range) and [number of batches analysed] for batch‐to‐batch variation and ranges for all other parameters.

	Belfeed B MP (solid)	Belfeed B ML (liquid)
Specifications 1
Xylanase activity (IU/g or mL)	≥ 400	≥ 400
Batch‐to‐batch variation	[5]	[5]
Xylanase activity (IU/g or mL)	408 (405–411)	405 (402–408)
Impurities	[3]	[3]
Lead (mg/kg)	< 0.050	< 0.050
Mercury (mg/kg)	< 0.005	< 0.005
Cadmium (mg/kg)	0.035–0.037	< 0.010
Arsenic (mg/kg)	< 0.10	< 0.10
Mycotoxins	[3]	[3]
Aflatoxins B1, B2, G1 and G2 (individual) (μg/kg)	< 0.1	< 0.1
Total aflatoxins (B1, B2, G1, G2) (μg/kg)	< 0.4	< 0.4
Ochratoxin A (μg/kg)	< 0.2	< 0.2
Deoxynivalenol (μg/kg)	< 20–22	< 20
Zearalenone (μg/kg)	< 10	< 10
Microbial contamination	[3]	[3]
Salmonella spp. (per 25g)	Not detected	Not detected
Escherichia coli (per 25g)	Not detected	Not detected
Bacillus cereus (CFU/g)	< 10–50	< 10
Enterobacteriaceae (CFU/g)	< 100–1800	< 100
Total coliforms (CFU/g)	< 10–3000	< 10
Yeast (CFU/g)	< 100	< 100
Moulds (CFU/g)	400	< 100
Physical properties
Dusting potential (mg/m3)	575–628 [3]	–
Bulk density (kg/m3)	799 [1]	–
pH	–	4.2 [5]
Viscosity (cP)	–	44.2–45.0 [3]
Density (g/mL)	–	1.04 (1.04–1.05) [3]

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 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. In addition, 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 Puratos NV2 for the renewal of the authorisation of the additive consisting of endo‐1,4‐beta‐xylanase produced with Bacillus subtilis LMG S‐15136 (Belfeed B MP/ML), when used as a feed additive for all poultry species, suckling and weaned piglets, pigs for fattening, lactating sows and all minor porcine species other than reproductive animals, and for the extension of the authorisation for use in feed for all porcine species (category: zootechnical additives; functional group: digestibility enhancers).

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 10 September 2024 and the general information and supporting documentation are available at https://open.efsa.europa.eu/questions/EFSA‐Q‐2024‐00563. The particulars and documents in support of the application were considered valid by EFSA as of 10 December 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 endo‐1,4‐beta xylanase produced with B. subtilis LMG S‐15136 (Belfeed B MP/ML), when used under the proposed conditions of use (see Section 3.1.3).

Additional information

1.2

The additive consists of endo‐1,4‐beta‐xylanase and is currently authorised for use in feed for poultry, weaned piglets and pigs for fattening;3 for suckling piglets and minor porcine species other than reproductive animals;4 and for lactating sows (4a1606i).5

EFSA issued four opinions on the safety and efficacy of this product when used in feed for poultry and porcine species (EFSA FEEDAP Panel, 2016, 2019a, 2019b, 2021a, 2021b, 2025).

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 endo‐1,4‐beta‐xylanase produced with B. subtilis LMG S‐15136 (Belfeed B MP/ML) as a feed additive.

The confidential version of the technical dossier was subject to a target consultation of the interested Member States from 13 December 2024 to 13 March 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 21 March to 11 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 or other expert bodies, peer‐reviewed scientific papers and experts' knowledge, 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 endo‐1,4‐beta‐xylanase produced with B. subtilis LMG‐S 15136 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 endo‐1,4‐beta‐xylanase produced with B. subtilis LMG‐S 15136 (Belfeed B MP/ML) 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, 2017a), Guidance on the identity, characterisation and conditions of use of feed additives (EFSA FEEDAP Panel, 2017b), Guidance on the assessment of the safety of feed additives for the target species (EFSA FEEDAP Panel, 2017c), 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, 2019a, 2019b), Guidance on the renewal of the authorisation of feed additives (EFSA FEEDAP Panel, 2021a, 2021b), 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, 2024) and EFSA statement on the requirements for whole genome sequence analysis of microorganisms intentionally used in the food chain (EFSA, 2024).

ASSESSMENT

3

The additive Belfeed B MP/ML (endo‐1,4‐beta‐xylanase (EC 3.2.1.8; xylanase) produced with a genetically modified strain of B. subtilis LMG S‐15136 is currently authorised as a zootechnical additive (functional group: digestibility enhancers) for use in feed for all poultry species, suckling and weaned piglets, pigs for fattening, lactating sows and minor porcine species other than reproductive animals. The applicant is requesting the renewal of the above authorisations and the extension of the authorisation to cover all porcine species/categories.

Characterisation

3.1

Characterisation of the production organism

3.1.1

The endo‐1,4‐beta‐xylanase is produced by fermentation with a genetically modified strain of B. subtilis which is deposited at the Belgian Coordinated Collections of Microorganisms (BCCM) with the accession number LMG S‐15136.11 The production strain and its genetic modifications were evaluated in a previous opinion (EFSA FEEDAP Panel, 2025) and considered of no concern. No changes have been introduced compared with the previous assessment. No additional data were provided in the current submission other than information on the potential antimicrobial activity (see Section 3.1.2).

Characterisation of the additive

3.1.2

The additive Belfeed B MP/ML is currently authorised as a preparation of endo‐1,4‐beta‐xylanase (EC 3.2.1.8; xylanase) produced with B. subtilis (LMG S‐15136). The additive is currently authorised in two formulations, solid (Belfeed B MP) and liquid (Belfeed B ML), both with a minimum xylanase activity of 400 IU12/g.

The applicant declares that no changes in the manufacturing process of the additive were implemented since the previous authorisations.13 Exception is ■■■■■. The information provided by the applicant supports the fact that the ■■■■■ steps included in the manufacturing process would likely prevent the presence of the ■■■■■ in the final product, although no data were provided to confirm this.

The data provided by the applicant on the batch‐to‐batch variation,14 and impurities15 of the additive are reported in Table 1. The data provided on physical and technological properties were already assessed in a previous opinion (, 2025; EFSA FEEDAP Panel, 2016) and are considered valid for the additive under assessment.16

The data provided by the applicant showed compliance with the specifications set in the authorising regulations for Belfeed B MP/ML. The Panel considers that the levels of the impurities analysed are of no concern. The Panel noted that the levels of microbial contamination in some/all batches (depending on the parameter considered) of the solid product are high and deserve attention/monitoring during the production process.

The presence of antimicrobial activity was tested in three batches of the intermediate product used to formulate the final additives.17 This was done using a disc‐diffusion agar method against the following reference strains: Staphylococcus aureus ATCC 6538, Escherichia coli ATCC 11229, Bacillus cereus ATCC 2, Niallia circulans ATCC 4516, Streptococcus pyogenes ATCC 12344 and Serratia marcescens ATCC 14041. No antimicrobial activity was detected.

The presence of viable cells of the production strain was investigated in three batches of the liquid enzyme concentrate (after sterile filtration) used to prepare the final formulations, analysed in triplicate (1 gram per sample).18 Methodology, incubation conditions and controls were performed according to the requirements set by the EFSA FEEDAP Panel (EFSA FEEDAP Panel, 2018). No viable cells of the production strain were found in any of the test samples.

The presence of DNA from the production strain was analysed by PCR in the same three batches described above, in triplicate (1 gram per sample).19 ■■■■■ The limit of detection ■■■■■ of the production strain was 10 ng per gram of product. No DNA from the production strain was detected in any of the samples.

Conditions of use

3.1.3

The additive is currently authorised for use in feed for all poultry, suckling and weaned piglets, pigs for fattening, lactating sows and all minor porcine species other than reproductive animals at a minimum inclusion level of 10 IU endo‐1,4‐beta‐xylanase/kg complete feed.

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

For all poultry species, weaned piglets and pigs for fattening

1. In the directions for use of the additive and premixture, indicate the storage conditions and stability to pelleting.
2. For use in weaned piglets up to 35 kg of body weight.
3. 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.

For suckling piglets and minor porcine species other than reproductive animals and lactating sows

1. In the directions for use of the additive and premixtures, the storage conditions and stability to heat treatment shall be indicated.
2. 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 and skin protection.

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

In addition, the applicant requested the extension of use to all porcine species/categories.

Safety

3.2

The safety aspects regarding the use of this additive in feed were previously assessed, including the safety of the genetic modification of the production strain, the safety for the target species, consumers, users and the environment (EFSA FEEDAP Panel, 2016, 2019a, 2019b, 2021a, 2021b, 2025).

In those assessments, the FEEDAP Panel concluded that the additive is safe for poultry species, and all Suidae species at the recommended use levels. The Panel also concluded that the additive is safe for the consumers and the environment. Regarding the safety for the user, the FEEDAP Panel concluded that both forms of the additive are irritant to skin and eyes and respiratory sensitisers. Any exposure is considered a risk (EFSA FEEDAP Panel, 2025).

The applicant stated that no adverse effect notifications were received since the previous authorisation with respect to the safety for target animals, consumers, users/workers or the environment.20

The applicant performed literature searches to provide evidence that in the light of the current knowledge the additive remains safe under the approved conditions for target species,21 consumers,22 users23 and the environment.24 The literature search covered the period from 2016 to 2024. The searches included at least two databases (Medline and Google Scholar for the target animals and consumers; plus ECHA Chem database and CompTox Chemicals Dashboard in the case of the safety for the user and the environment). No hits reporting adverse effects of the additive on target species, consumers, users or the environment were retrieved.

Considering the above and the fact that the manufacturing process and the composition have not been modified, the Panel concludes that the use of the additive under the approved conditions of the authorisation remains safe for the target animals, the consumers and the environment. Both forms of the additive are irritant to skin and eyes and respiratory sensitisers. Any exposure is considered a risk.

In the context of this application, the applicant requested for an extension of use to all Suidae. The FEEDAP Panel already concluded that the use of the additive in all Suidae is safe for the target species, consumers and the environment (EFSA FEEDAP Panel, 2019a, 2019b).

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 all poultry and for weaned piglets, pigs for fattening, suckling piglets, all minor porcine species other than reproductive animals and for lactating sows and weaned piglets. Therefore, there is no need to assess the efficacy of the additive in the context of the renewal of the authorisation.

The applicant is requesting an extension of use to all Suidae at the currently authorised conditions of use. The efficacy of the additive in gestating sows has been established in a recent opinion (EFSA FEEDAP Panel, 2025). Therefore, considering that the efficacy has been demonstrated in growing and reproductive pigs, the Panel concludes that the additive, in both forms, is efficacious in all porcine species/categories at the minimum use level of 10 IU/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 Regulation25 and Good Manufacturing Practice.

CONCLUSIONS

4

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

The FEEDAP Panel concludes that the additive Belfeed B MP/ML remains safe for the target species, the consumer and the environment under the approved conditions of the authorisations. The extension of use to all porcine species would not affect these conclusions; therefore, the additive is considered safe for the target species, the consumer and the environment.

Regarding the safety for the user, the two formulations of the additive are considered skin and eye irritants, and respiratory sensitisers. Any exposure is considered a risk.

There is no need for assessing the efficacy of the additive in the context of the renewal of the authorisation. The FEEDAP Panel concludes that the additive is efficacious in all porcine species/categories at 10 IU endo‐1,4‐beta‐xylanase/kg complete feed.

RECOMMENDATIONS

5

The Panel notes that the analytical values found for microbial contamination in Belfeed B MP are high and deserve attention/monitoring during the production process.

ABBREVIATIONSBCCMBelgian Coordinated Collections of MicroorganismsCASChemical Abstracts ServiceCFUcolony forming unitDMdry matterEINECSEuropean Inventory of Existing Chemical SubstancesEURLEuropean Union Reference LaboratoryFEEDAPEFSA Scientific Panel on Additives and Products or Substances used in Animal FeedIUInternational unitIUPACInternational Union of Pure and Applied Chemistry

REQUESTOR

European Commission

QUESTION NUMBER

EFSA‐Q‐2024‐00563

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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, Katerina Theodoridou

LEGAL NOTICE

Relevant information or parts of this scientific output have been blackened in accordance with the confidentiality requests formulated by the applicant pending a decision thereon by EFSA. The full output has been shared with the European Commission, EU Member States (if applicable) and the applicant. The blackening may be subject to review once the decision on the confidentiality requests is adopted by EFSA and in case it rejects some of the confidentiality requests