Safety and efficacy of a feed additive consisting of Saccharomyces cerevisiaeMUCL 39885 (Biosprint®) for all pigs (other than sows and weaned piglets) and other minor porcine species (Prosol S.p.A.).

The additive Biosprint® contains viable cells of Saccharomyces cerevisiae MUCL 39885 and is authorised as a feed additive in sows, dairy cows, horses, piglets (weaned), cattle for fattening, minor ruminants for fattening and minor ruminants for dairy products. The applicant has requested to extend the use of the additive to all pigs (other than sows and weaned piglets) and other minor porcine species at a minimum inclusion level of 3 × 109 CFU/kg feed. S. cerevisiae is considered by EFSA to have qualified presumption of safety (QPS) status and consequently is considered safe for the target species, the consumers and the environment. The additive is considered as a potential skin and eye irritant and a skin and respiratory sensitiser. In previous evaluations, the FEEDAP Panel concluded that the additive is efficacious in sows and weaned piglets. In the current application, these conclusions are extrapolated to all pigs (other than sows and weaned piglets) and to other minor porcine species.

Introduction

Background and Terms of Reference as provided by the requestor

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 a feed additive shall submit an application in accordance with Article 7.

The European Commission received a request from Prosol S.p.A.2 for authorisation of the preparation of Saccharomyces cerevisiae MUCL 39885 (Biosprint®), when used as a feed additive for all pigs (other than sows, suckling and weaned piglets) and other minor porcine species (category: zootechnical additives; functional group: gut flora stabiliser). During the assessment, the applicant requested a change in the species from ‘for all pigs (other than sows, suckling and weaned piglets) and other minor porcine species’ to ‘for all pigs (other than sows and weaned piglets) and other minor porcine species.

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). The particulars and documents in support of the application were considered valid by EFSA as of 8 January 2021.

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 environment and on the efficacy of the preparation of S. cerevisiae MUCL 39885 (Biosprint®) when used under the proposed conditions of use (see Section 3.1.2).

Additional information

This additive is a preparation containing viable cells of S. cerevisiae MUCL 39885. It is currently authorised in sows,3 dairy cows, horses,4 piglets (weaned),5 cattle for fattening,6 minor ruminants for fattening and minor ruminants for dairy products (4b1710).7

The EFSA FEEDAP Panel issued several opinions on the safety and efficacy of Biosprint® (S. cerevisiae MUCL 39885) in different target species (EFSA, 2004, 2009, 2010a,b,c; EFSA FEEDAP Panel, 2013a, 2015, 2019a,b, 2020).

Data and methodologies

Data

The present assessment is based on data submitted by the applicant in the form of a technical dossier8 in support of the authorisation request for the use of Biosprint® (Saccharomyces cerevisiae MUCL 39885) as a feed additive.

The European Union Reference Laboratory (EURL) considered that the conclusions and recommendations reached in the previous assessment are valid and applicable for the current application.9

Methodologies

The approach followed by the FEEDAP Panel to assess the safety and the efficacy of Biosprint® (Saccharomyces cerevisiae MUCL 39885) is in line with the principles laid down in Guidance on the identity, characterisation and conditions of use of feed additives (EFSA FEEDAP Panel, 2017), Guidance on the characterisation of microorganisms used as feed additives or as production organisms (EFSA FEEDAP Panel, 2018a) and Guidance on the assessment of the efficacy of feed additives (EFSA FEEDAP Panel, 2018b).

Assessment

The additive contains viable cells of S. cerevisiae MUCL 39885. It is proposed to be used as a zootechnical additive (functional group: gut flora stabiliser) in feed for all pigs (other than sows and weaned piglets) and other minor porcine species.

The additive is currently authorised for use in sows and weaned piglets; the applicant is asking for the extension of the authorisation to all pigs and to all minor porcine species.

Characterisation

The additive contains viable cells of the strain S. cerevisiae MUCL 39885 (no carriers or excipients are present in the final product) and it is authorised with a minimum declared content of 1 × 109 colony forming unit (CFU)/g. The applicant proposes the same specifications as those currently authorised.

According to the applicant, no modifications have been introduced in the manufacturing process along the last nine years (EFSA FEEDAP Panel, 2010a,b,c).

The additive is marketed in two forms, ‘spherical’ (Biosprint® S) and ‘granulated’ (Biosprint® G). Compliance with the specifications set in the authorisation was confirmed by analysis of five batches of the G form (range 1.62–1.79 × 1010, mean 1.70 × 1010 CFU/g) and five batches of the S form (range 1.69–1.77 × 1010, mean 1.74 × 1010 CFU/g).

The same batches of the additive were analysed for microbial contamination. The results confirm compliance with limit levels set (Escherichia coli < 10 CFU/g, Salmonella spp. no detection in 25 g, filamentous fungi < 10 CFU/g, Listeria monocytogenes no detection in 1 g, coliforms < 100 CFU/g, Staphylococcus aureus < 10 CFU/g).10

The possible presence of chemical contaminants was measured on three recent batches of the product. In all the three batches, the concentration of heavy metals (cadmium, lead, mercury) and arsenic were below the corresponding limits of quantification (LOQs).11 Levels of chromium (three batches below the LOQ), cobalt (two batches were below the LOQ – 0.001 mg/kg; one batch measured 0.36 mg/kg), iron (range 5.89–49.5 mg/kg), nickel (two batches were below the LOQ – 0.0013 mg/kg; one batch measured 0.09 mg/kg), copper (range 3.7–7.92 mg/kg) and zinc (range 83.5–152 mg/kg) were also investigated. In the same batches, levels of aflatoxins B1, B2, G1 and G2 were < LOQ (0.3 or 0.5 mg/kg), of deoxynivalenol < 20 μg/kg (analysed in two batches only), of ochratoxin A < 1 μg/kg (analysed in two batches only), of zearalenone < 10 μg/kg (analysed in two batches only). Levels of pesticides screened in a multiresidue analysis were < 0.01 mg/kg, with the exception of 2,4,6‐trichlorophenol (0.04 mg/kg), epoxiconazole (0.076 mg/kg), tetraconazole (0.018 mg/kg) and difenoconazole (0.025 mg/kg) in one batch.12 Based on the results, no concern is identified.

The same batches of the G form of the additive were tested in triplicate for dusting potential13 according to the Stauber–Heubach method. The average value of dust was 260 mg/m3 (range: 217–290 mg/m3). The particle size distribution measured by sieving of three batches of the product for both formulations was provided. Regarding the S form, most of the particles had a diameter > 355 μm, with no particles below 125 μm. The G form of the additive had most of the particles with a diameter > 125 μm, with no particles below 90 μm.

The non‐genetically modified strain of S. cerevisiae composing the additive is deposited in the Belgian Coordinated Collection of Microorganism Culture Collection – Mycothéque de l'Université Catholique de Louvain (BCCM/MUCL) with the accession number 39885.14

A phylogenetic analysis based on ribosomal and mitochondrial protein sequences deduced from whole genome sequence data confirmed the taxonomic identification of the strain as S. cerevisiae. The analysis of the single nucleotide polymorphisms (SNP) in the whole genome as compared the genome of S. cerevisiae S288C and other 6 Saccharomyces species was also submitted and supports the identification of the BCCM/MUCL 39885 strain as S. cerevisiae.15

Stability and homogeneity

Stability and homogeneity of Biosprint® has been previously assessed (EFSA FEEDAP Panel, 2010a,b,c). In the current dossier, the applicant has provided a new shelf‐life study carried out in one batch of the additive stored at 25°C for 24 months, in aluminium bags under vacuum.16 At the end of the storage period, the S. cerevisiae counts losses were below 0.5 log.

Conditions of use

The additive is intended to be used in feed for all pigs (other than sows and weaned piglets) and other minor porcine species at a minimum inclusion level of 3 × 109 CFU/kg feed.

Safety

The species S. cerevisiae is considered by EFSA to be suitable for the Qualified Presumption of Safety (QPS) approach to safety assessment (EFSA, 2007, EFSA BIOHAZ Panel, 2020). This approach requires the identity of the strain under assessment to be conclusively established. In the context of the current application, the identity of the active agent was confirmed as S. cerevisiae. Accordingly, this strain is presumed safe for the target species, the consumer and the environment.

The safety for the user has been evaluated in a previous opinion (EFSA, 2010a,b,c). In 2010, the Panel concluded that the additive should be considered as a potential skin and eye irritant and a skin sensitiser and that the inhalation exposure would be minimal. No additional data were provided in the current application that would lead to a revision of the previous conclusions. Considering the proteinaceous nature of the additive, Biosprint® should be considered a respiratory sensitiser.

The applicant in support of the safety of the additive has submitted results from several literature searches.

Three literature searches on the safety of the additive for different target species, consumer, environment and user, covering the periods 2008–2018, 2018–2019 and 2019–March 2020, were evaluated by EFSA FEEDAP Panel in previous assessments (EFSA FEEDAP Panel, 2019a,b, 2020). These searches did not reveal any safety issue related to the additive under assessment.

The applicant performed two new literature searches on the safety of the additive for pigs and dogs, covering the periods 2010–June 2020 and 2010–July 2020, respectively, using the following databases: CAB Abstracts, PubMed and Scopus. It included ‘Saccharomyces cerevisiae’ and other terms relevant for target species safety and for toxicological aspects. This new search retrieved 7 publications, all of them were EFSA scientific opinions regarding the use of Saccharomyces cerevisiae as a feed additive.

No other relevant papers were identified that would highlight a safety concern for the target species, the consumer or the environment. Moreover, no relevant papers were identified that would add any additional concerns to those already identified for the safety for the user.

Conclusions of safety

The FEEDAP Panel concludes that Biosprint® in any of its forms is considered safe for the target species, for the consumer, and the environment. The additive should be considered as a potential skin and eye irritant and a skin and respiratory sensitiser.

Efficacy

In the current application dossier, the applicant is requesting to extend the use of the additive to all pigs (other than sows and weaned piglets) and other minor porcine species.

EFSA issued two opinions concluding that the additive is efficacious in sows (at a minimum content of 6.4 × 109 CFU/kg feed) (EFSA, 2009) and in weaned piglets (at a minimum content of 4 × 109 CFU/kg feed) (EFSA, 2010c).

Considering that efficacy has been shown in weaned piglets and in sows, the Panel considers that these conclusions can be extended/extrapolated to all pigs and other minor porcine species. Therefore, the FEEDAP Panel concludes that the additive is efficacious in all growing pigs at the level of 4 × 109 CFU/kg feed, in all reproductive animals at 6.4 × 109 CFU/kg feed and in all other minor porcine species at the respective levels: growing 4 × 109 CFU/kg feed and reproductive at 6.4 × 109 CFU/kg feed.

The FEEDAP Panel noted that the applicant has requested an inclusion level lower than those indicated above (3 × 109 CFU/kg feed). However, in the absence of data, the FEEDAP Panel was not able to conclude on the efficacy of the additive when administered at the requested inclusion level.

Post‐market monitoring

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 Regulation17 and Good Manufacturing Practice.

Conclusions

Biosprint® is considered safe for the target species, consumers and the environment.

The additive is considered as a potential skin and eye irritant and a skin and respiratory sensitiser.

The FEEDAP Panel concludes that the additive has the potential to be efficacious (gut flora stabiliser) in all growing pigs at the level of 4 × 109 CFU/kg feed, in all reproductive pigs at 6.4 × 109 CFU/kg feed and in all other minor porcine species at the respective levels: growing at 4 × 109 CFU/kg feed and reproductive at 6.4 × 109 CFU/kg feed.

In the absence of data, the FEEDAP Panel is not able to conclude on the efficacy of the additive when administered at the requested inclusion level (3 × 109 CFU/kg feed).

Documentation as provided to EFSA/Chronology

Abbreviations

Belgian Coordinated Collection of Microorganism Culture Collection ‐ Mycothéque de l'Université Catholique de Louvain

colony forming unit

European Union Reference Laboratory

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

Mycothéque de l'Université Catholique de Louvain

limit of quantification

Qualified Presumption of Safety

single nucleotide polymorphism

whole genome sequence

Date	Event
12/08/2020	Reception of the Evaluation report of the European Union Reference Laboratory for Feed Additives
29/08/2020	Dossier received by EFSA. Biosprint® for all pigs (other than sows, suckling and weaned piglets) and other minor porcine species. Submitted by Prosol S.p.A.
07/09/2020	Reception mandate from the European Commission
08/01/2021	Application validated by EFSA – Start of the scientific assessment
08/04/2021	Request of supplementary information to the applicant in line with Article 8(1)(2) of Regulation (EC) No 1831/2003 – Scientific assessment suspended. Issues: characterisation
09/04/2021	Comments received from Member States
13/04/2021	Reception of supplementary information from the applicant ‐ Scientific assessment re‐started
23/06/2021	Opinion adopted by the FEEDAP Panel. End of the Scientific assessment