Safety and efficacy of a feed additive consisting of Bacillus velezensis DSM 15544 (Calsporin®) for dairy cows and other dairy ruminants (Asahi Biocycle Co. Ltd.).

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 Calsporin® (Bacillus velezensis DSM 15544) as a zootechnical additive for dairy cows and other dairy ruminants. The product under assessment is based on viable spores of Bacillus velezensis DSM 15544. B. velezensis is considered suitable for the qualified presumption of safety (QPS) approach to safety assessment. The identity of the active agent was established and compliance with the applicable qualifications confirmed. Therefore, B. velezensis DSM 15544 is presumed safe for the target species, consumers of products derived from animals fed the additive and the environment. Since no concerns are expected from the other components of the additive, Calsporin® is also considered safe for the target species, consumers of products derived from animals fed the additive and the environment. The additive is not a dermal/eye irritant or a skin sensitiser but should be considered a respiratory sensitiser. The FEEDAP Panel is not in the position to conclude on the efficacy of Calsporin® for dairy cows or other dairy ruminants based on the data provided.

Introduction

Background and Terms of Reference

Regulation (EC) No 1831/2003 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.

The European Commission received a request from Asahi Biocycle Co. Ltd., Japan, represented in the EU by Pen & Tec Consulting SLU for the authorisation of the additive consisting of Bacillus velezensis DSM 15544 (Calsporin®) when used as a feed additive for dairy cows and other dairy ruminants (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). The particulars and documents in support for the application were considered valid by EFSA as of 12 May 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 the environment and on the efficacy of the feed additive consisting of Bacillus velezensis DSM 15544 (Calsporin®), when used under the proposed conditions of use (see Section 3.1.2).

Additional information

The subject of the assessment is the feed additive consisting of viable spores of a strain of Bacillus velezensis (DSM 15544), intended for use as a zootechnical additive (functional group: gut flora stabiliser) for dairy cows and other dairy ruminants.

The European Food Safety Authority (EFSA) has issued several opinions on the safety and efficacy of Calsporin® as a feed additive for different species: chickens for fattening (EFSA, 2006, 2007a; EFSA FEEDAP Panel, 2018a), weaned piglets (EFSA FEEDAP Panel, 2010a, 2020), turkeys for fattening, ducks, geese, pigeons and other game birds for meat production, ducks, geese, pigeons, game birds, ornamental and sporting birds for rearing to point of lay, turkeys reared for breeding and chickens reared for laying (EFSA FEEDAP Panel, 2010b), laying hens and avian species for laying (EFSA FEEDAP Panel, 2015a), ornamental fish (EFSA FEEDAP Panel, 2015b), sows and suckling piglets (EFSA FEEDAP Panel, 2017a), dogs (EFSA FEEDAP Panel, 2017b), pigs for fattening (EFSA FEEDAP Panel, 2018b), all poultry species (EFSA FEEDAP Panel, 2019a) and for piglets (suckling and weaned), pigs for fattening, sows in order to have benefit in piglets, ornamental fish, dogs and all avian species (EFSA FEEDAP Panel, 2021).

The additive is authorised in the European Union (EU) as a zootechnical additive (functional group: gut flora stabiliser) for use in feed for chickens for fattening, weaned piglets, chickens reared for laying, turkeys, minor avian species and other ornamental and game birds, laying hens and ornamental fish, dogs, suckling piglets and sows and pigs for fattening (4b1820).

Data and methodologies

Data

The present assessment is based on data submitted by the applicant in the form of a technical dossier in support for the authorisation request for the use of Calsporin® (B. velezensis DSM 15544) as a feed additive.

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 active agent in animal feed are valid and applicable for the current application.

Methodologies

The approach followed by the FEEDAP Panel to assess the safety and the efficacy of B. velezensis DSM 15544 (Calsporin®) is in line with the principles laid down in Regulation (EC) No 429/2008 and the relevant guidance documents: Guidance on studies concerning the safety of use of the additive for users/workers (EFSA FEEDAP Panel, 2012); Guidance on the assessment of the safety of feed additives for the consumer (EFSA FEEDAP Panel, 2017c); Guidance on the identity, characterisation and conditions of use of feed additives (EFSA FEEEDAP Panel, 2017d); Guidance on the assessment of the safety of feed additives for the target species (EFSA FEEDAP Panel, 2017e); Guidance on the assessment of the efficacy of feed additives (EFSA FEEDAP Panel, 2018c); Guidance on the characterisation of microorganisms used as feed additives or as production organisms (EFSA FEEDAP Panel, 2018d); Guidance on the assessment of the safety of feed additives for the environment (EFSA FEEDAP Panel, 2019b).

Assessment

The subject of the assessment is a product containing viable spores of a single strain of Bacillus velezensis intended for use as a zootechnical additive (functional group: gut flora stabiliser) in feed for dairy cows and other dairy ruminants. The additive will be hereafter referred to as Calsporin® (its tradename).

Characterisation

Characterisation of the additive

Calsporin® is a powder with a minimum declared content of 1 × 1010 colony forming units (CFU) of B. velezensis DSM 15544 per gram of additive.

It has the same formulation (■■■■■ and method of manufacture as that considered in the most recent opinion adopted by the FEEDAP Panel in 2020. Thus, the data pertaining to composition, impurities, physical properties and shelf‐life described in that opinion (EFSA FEEDAP Panel, 2020) apply to the current assessment.

The batch‐to‐batch variation of seven batches of the additive showed compliance with the minimum specifications (mean 1.2 × 1010 CFU/g, with a range of 1.1–1.3 × 1010 CFU/g).

In that same opinion, the active agent originally identified as Bacillus subtilis was reclassified as B. velezensis and fully characterised as per the requirements of the FEEDAP guidance on the characterisation of microorganisms used as feed additives or as production organisms.

The stability and homogeneous distribution of the additive when mixed with premixtures for poultry and pigs were also established in previous opinions (EFSA FEEDAP Panel, 2010a,b, 2018a). The FEEDAP Panel is of the opinion that these existing data are sufficient to establish the homogeneity of Calsporin® when mixed in premixtures for the new target species for which authorisation is sought.

No data have been produced to support the stability and capacity of the additive to homogeneously disperse in feed for dairy ruminants.

Conditions of use

The additive is intended for use in feed for dairy cows and other dairy ruminants at a proposed minimum daily dose of 3 × 109 CFU/head. No inclusion level in CFU/kg feed was provided.

Safety

The species B. velezensis is considered by EFSA to be suitable for the qualified presumption of safety (QPS) approach to safety assessment (EFSA, 2007b; EFSA BIOHAZ Panel, 2021). This approach requires the identity of the strain to be conclusively established and evidence provided that it does not show acquired resistance to relevant antimicrobials, that it lacks toxigenic potential and that it does not produce aminoglycosides. In a previous opinion (EFSA FEEDAP Panel, 2020), the identity of the active agent was established and the compliance with the other applicable qualifications confirmed. Accordingly, B. velezensis DSM 15544 was presumed safe for the target species, consumers of products derived from animals fed the additive and the environment. Since no concerns are expected from the other components of the additive, Calsporin® was also presumed safe for the target species, consumers and the environment.

The applicant conducted a trial aimed to evaluate the possible presence of B. velezensis DSM 15544 in milk from dairy cows fed diets supplemented with Calsporin®. For this purpose, 40 lactating dairy cows were divided into two groups and fed the same basal total mixed ration plus a concentrate not supplemented (control), or supplemented with Calsporin® to reach a daily dose of 6 × 109 CFU/head. Cows were under study for 28 days and milk and faecal samples were collected and analysed for Bacillus spp. and B. velezensis DSM 15544 counts. Milk and faecal samples from 10 animals per treatment were collected at different time points (0, 14 and 28 days) and samples from the milk tanks (one for control and one for Calsporin group) were obtained on days 0, 7, 14, 21 and 28. No significant differences in Bacillus spp. counts were observed between treatment groups in any type of sample. No B. velezensis DSM 15544 colonies were detected in milk samples from the Calsporin® or control group. The cows receiving the additive had higher counts of B. velezensis DSM 15544 in faeces compared to control. These results may indicate that the exposure of the consumers to the active agent under assessment in milk from cows receiving the additive is unlikely.

The safety for the users was evaluated by the FEEDAP Panel in a previous opinion (EFSA FEEDAP Panel, 2018a). The Panel concluded that the additive is not a dermal/eye irritant or a skin sensitiser but should be considered a respiratory sensitiser. No additional data were provided in the current application. The use of the additive in dairy cows and other dairy ruminants is considered unlikely to introduce hazards for users of the product not already considered as part of the previous assessment. Therefore, the conclusions reached in the previous assessment apply to the current application.

Efficacy

Efficacy for dairy cows

Three efficacy studies with dairy cows were submitted. However, none could be further considered for the assessment due to substantial flaws in the experimental design and reporting, including: housing and feeding conditions not fully reported (study 1), feed intake not (studies 1 and 3 ) or insufficiently (study 2) recorded/reported, experimental groups not comparable at baseline on differences in days in milk and number of lactations (study 2), non‐compliance with guidance provision that efficacy studies in dairy cows should start at 4–8 weeks after calving (studies 2 and 3) or the information at that regard is not reported (study 1).

Therefore, the Panel is not in the position to conclude on the efficacy of Calsporin® for dairy cows based on the data provided.

Efficacy for other dairy ruminants

In the absence of evidence of efficacy in dairy cows, no conclusions can be drawn on the efficacy of Calsporin® for other dairy ruminants.

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 Regulation and Good Manufacturing Practice.

Conclusions

Calsporin® is considered safe for the target species, consumers of products derived from animals fed the additive and the environment.

Calsporin® is not a dermal/eye irritant or a skin sensitiser but should be considered a respiratory sensitiser.

The FEEDAP Panel is not in the position to conclude on the efficacy of Calsporin® for dairy cows or other dairy ruminants based on the data provided.

Documentation provided to EFSA/Chronology

colony‐forming unit

European Union Reference Laboratory

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

Date	Event
15/03/2021	Dossier received by EFSA. CALSPORIN® Bacillus velezensis DSM 15544. Zootechnical feed additive for dairy cows and other dairy ruminants, functional group: gut flora stabiliser. Submitted by Asahi Biocycle Co. Ltd represented by Pen & Tec Consulting SLU
23/03/2021	Reception mandate from the European Commission
12/05/2021	Application validated by EFSA – Start of the scientific assessment
28/09/2021	Comments received from Member States
10/11/2021	Opinion adopted by the FEEDAP Panel. End of the Scientific assessment