Safety and efficacy of a feed additive consisting of Bacillus paralicheniformis DSM 33902 and Bacillus subtilis DSM 33903 (Bovacillus™) for dairy cows and other dairy ruminants (Chr. Hansen A/S)

Abstract

Following a request from the European Commission, EFSA was asked to deliver a scientific opinion on the safety and efficacy of Bovacillus™, a product containing viable cells of Bacillus paralicheniformis DSM 33902 and Bacillus subtilis DSM 33903, which is intended to be used as a zootechnical additive (functional group: gut flora stabiliser) in the feed of dairy cows and other dairy ruminants. The trade name is Bovacillus™, and it is intended to be commercialised in two formulations: Bovacillus™ 10, to use in feed, and WS, to use in drinking water. B. paralicheniformis and B. subtilis are considered suitable for the qualified presumption of safety (QPS) approach to safety assessment. The identity of the active agents was established, and compliance with the applicable qualifications confirmed. Therefore, Bacillus paralicheniformis DSM 33902 and Bacillus subtilis DSM 33903 are 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, Bovacillus™ is also considered safe for the target species, consumers and the environment. Bovacillus™ 10 and WS are not skin or eye irritants. Due to the nature of the active agents, both forms of the additive are considered respiratory and skin sensitisers. The Panel concludes that Bovacillus™ has the potential to be efficacious as a zootechnical additive when supplemented in the feed of dairy ruminants at 3.8 × 10⁸ CFU/kg complete feed.

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 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 Chr. Hansen A/S ² for the authorisation of the additive consisting of Bacillus paralicheniformis DSM 33902 and Bacillus subtilis DSM 33903 (Bovacillus™), when used as a feed additive for dairy cows and other dairy ruminants (category: zootechnical additives; functional group: gut flora stabiliser).

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 dossier was received on 30 June 2023 and the general information and supporting documentation are available at https://open.efsa.europa.eu/questions/EFSA‐Q‐2023‐00454. The particulars and documents in support of the application were considered valid by EFSA as of 28 September 2023.

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 B. paralicheniformis DSM 33902 and B. subtilis DSM 33903 (Bovacillus™), when used under the proposed conditions of use (see Section 3.1.4 ).

1.2. Additional information

The additive is a preparation containing Bacillus paralicheniformis DSM 33902 and Bacillus subtilis DSM 33903 (Bovacillus™). It has not been previously authorised as a feed additive for dairy ruminants in the European Union.

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 ³ in support of the authorisation request for the use of the preparation containing B. paralicheniformis DSM 33902 and Bacillus subtilis DSM 33903 (Bovacillus™) as a feed additive.

The confidential version of the technical dossier was subject to a target consultation of the interested Member States from 28 September 2023 to 28 December 2023, for which the received comments were considered for the assessment.

In accordance with Article 38 of the Regulation (EC) No 178/2002 ⁴ 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, ⁵ 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 3 April to 24 April 2024 for which no comments were received.

EFSA has verified the European Union Reference Laboratory (EURL) report as it relates to the methods used for the control of the active substances in animal feed. ⁶

2.2. Methodologies

The approach followed by the FEEDAP Panel to assess the safety and the efficacy of B. paralicheniformis DSM 33902 and B. subtilis DSM 33903 (Bovacillus™) is in line with the principles laid down in Regulation (EC) No 429/2008 ⁷ 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 FEEEDAP 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, 2019), EFSA statement on the requirements for whole genome sequence analysis of microorganisms intentionally used in the food chain (EFSA, 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, 2024a, 2024b).

3. ASSESSMENT

The additive consists of viable spores of B. paralicheniformis DSM 33902 and B. subtilis DSM 33903 and is intended to be used as a zootechnical additive (functional group: gut flora stabilisers) in feed and water for dairy cows and dairy ruminants. The additive will be referred to in this opinion with its trade name Bovacillus™.

3.1. Characterisation

3.1.1. Characterisation of the active agents

B. paralicheniformis DSM 33902 was isolated from soil, and B. subtilis DSM 33903 from soybean mash. Both were deposited at the Deutsche Sammlung von Mikroorganismen und Zellkulturen. ⁸ According to the applicant, none of the strains has been genetically modified.

The taxonomic classification of both strains was established by bioinformatic analysis of whole genome sequence data (WGS). ⁹ The identification of the DSM 33902 strain was achieved by average nucleotide identity (ANI) calculation of its WGS data and the genome sequences of type strains of the species in the Bacillus genus, showing an ANI value of 98.6% with that of the type strain B. paralicheniformis KJ‐16^T. This was in line with the results of the comparison of the sequences of gyrA and rpoB genes retrieved from the WGS of the DSM 33902 strain, which yielded identity percentages above 99.6% with the respective gene sequences retrieved from the type strain genome.

For the DSM 33903 strain, an ANI value of 97.7% was attained for the comparison between its genome sequence and that of B. subtilis 168^T. This result was in line with the results of the comparison of sequences of groEL, gyrA, polC, purH and rpoB genes retrieved from the WGS of the DSM 33903 strain, which showed identity percentages above 98.6% with the respective gene sequences retrieved from the type strain genome.

The susceptibility of the two bacterial strains was tested against the battery of antibiotics recommended by the FEEDAP Panel by broth microdilution (EFSA FEEDAP Panel, 2018). ¹⁰ For B. subtilis DSM 33903, all the minimum inhibitory concentration (MIC) values fell below the corresponding cut‐off values for Bacillus spp. Therefore, B. subtilis DSM 33903 is susceptible to all the antibiotics tested. The MIC values for B. paralicheniformis DSM 33902 fell below the corresponding cut‐off values for Bacillus spp., except for chloramphenicol, clindamycin and erythromycin. The MIC values for the first two were one dilution above the corresponding cut‐off values. Exceeding the cut‐off value by one dilution is considered within the normal range of variation and, thus, is not a matter of concern. The MIC value for erythromycin was at least two dilutions above the corresponding cut‐off value (MIC > 16 mg/L, cut‐off: 4 mg/L). Therefore, the strain is susceptible to the relevant antibiotics, except for erythromycin.

The WGS data of both strains were interrogated for the presence of antimicrobial resistance (AMR) genes by a search against the ResFinder database and the NCBI Bacterial Antimicrobial Resistance Reference Gene database. ¹¹ A total of four and two hits exceeding the thresholds recommended by EFSA (EFSA, 2021) were found in the genomes of the DSM 33903 and DSM 33902 strains, respectively. Further analyses of these hits following EFSA's criteria (EFSA BIOHAZ Panel, 2023) did not reveal any acquired AMR genes. Therefore, the FEEDAP Panel concludes that the strains harbour no acquired AMR genes and raise no safety concerns. Although the DSM 33902 strain was resistant to erythromycin, since no acquired AMR genes were found in the WGS, the resistance to erythromycin does not raise safety concerns.

A cytotoxicity test with Vero cells for each strain was conducted according to the above‐mentioned FEEDAP guidance (EFSA FEEDAP Panel, 2018). ¹² No lysis of Vero cells was detected; therefore, the strains are considered non‐toxigenic.

To exclude the capacity of B. paralicheniformis DSM 33902 to produce bacitracin, the supernatants from the culture were tested against reference strains. ¹³ No inhibition by DSM 33902 was observed in any of the reference strains, denoting the inability of bacitracin production at any levels of concern for the intended use.

3.1.2. Characterisation of the additive

Each of the Bacillus spp. strains is grown separately. After sporulation, the viable spores of B. paralicheniformis are pasteurised. The viable spores of both strains are later harvested by centrifugation, and then the liquid concentrate is spray‐dried. Once dried, the two Bacillus concentrates are blended on a colony‐forming units (CFU) basis with the corresponding carrier, depending on the final formulation of the additive: calcium carbonate for Bovacillus™ 10 and maltodextrin for Bovacillus™ WS. The carrier represents approximately ■■■■■ (w/w) of the final additive in both formulations.

The applicant declared that no antimicrobial products are used in the manufacturing process.

The guaranteed minimum total concentration of viable spores of Bacillus spp. in the product is 3.2 × 10¹⁰ CFU/g additive, with a 1:1 ratio for each of the strains. Compliance was demonstrated in five batches of each formulation, showing Bovacillus™ 10 an average content of total bacilli of 4.6 × 10¹⁰ (4.3–4.9 × 10¹⁰) CFU/g product, with an average count of 2.3 (2.1–2.4) × 10¹⁰ CFU/g product for B. subtilis and 2.4 (2.2–2.5) × 10¹⁰ CFU/g for B. paralicheniformis; and Bovacillus™ WS an average content of 4.3 × 10¹⁰ (4.1–4.5 × 10¹⁰) CFU/g, with an average count of 2.1 (2.0–2.1) × 10¹⁰ CFU/g product for B. subtilis and 2.2 (2.1–2.4) × 10¹⁰ CFU/g for B. paralicheniformis. ¹⁴

The applicant established specifications for the presence of arsenic (< 3 mg/kg), cadmium (< 0.5 mg/kg), mercury (< 0.5 mg/kg), lead (< 5 mg/kg) and aflatoxin B1 (< 0.01 mg/kg). Three batches of each formulation were analysed. ¹⁵ The batches of Bovacillus™ 10 showed mean concentrations of 0.48 (0.46–0.49) mg arsenic/kg, 0.77 (0.74–0.80) mg lead/kg, 0.13 (0.13–0.13) mg cadmium/kg and 0.004 (0.003–0.005) mg mercury/kg, while aflatoxin B1 was below the limit of quantification (LOQ = 0.46 μg/kg) in all cases. In all batches of Bovacillus™ WS, cadmium and aflatoxin B₁ were below the LOQ, while arsenic (0.012 mg/kg) and lead (0.024 mg/kg) were only detected in one of the batches analysed; the mean concentration of mercury was 0.0010 (0.0008–0.0012) mg/kg.

Microbiological contamination was analysed in five batches of each formulation. In all batches analysed, the results showed values of Escherichia coli < 10 CFU/g of additive, coliforms < 1000 CFU/g, yeasts and moulds < 1000 CFU/g and no detection of Salmonella spp. in 25 g. ¹⁶ These values were compliant with the specifications set by the applicant. Additionally, the applicant analysed the counts of Enterobacteriaceae ¹⁷ and Bacillus cereus ¹⁸ in three batches of each formulation, with values < 10 CFU/g in all cases.

The FEEDAP Panel considers that the amounts of detected impurities do not raise safety concerns.

Both formulations are presented as white solid preparations. The bulk density (average of three batches) is 1341 and 546 kg/m3 for Bovacillus™ 10 and WS, respectively, and the tapped density (average of three batches) is 1595 and 702 kg/m³ for Bovacillus™ 10 and WS, respectively. ¹⁹ The dusting potential of three batches of each formulation of the additive was determined using the Stauber‐Heubach method and showed values on average of 1886 mg/m³ (943–2480 mg/m³) and 1149 mg/m³ (581.3–2226 mg/m³) for Bovacillus™ 10 and WS, respectively. ²⁰ The particle size of the additive was analysed by laser‐diffraction method; the results showed that, for Bovacillus™ 10, 13.0, 9.1, and 6.9% of the particles of the additive showed a diameter < 100 μm, < 50 μm and < 10 μm, respectively. For Bovacillus™ WS, the respective values were 60.0, 24.1 and 2.70%. ²¹

3.1.3. Stability and homogeneity

The shelf‐life of the additive was determined by monitoring samples of three batches of each formulation stored in a sealed light‐ and air‐tight packaging at 25 (both formulations) and 37°C (Bovacillus™ 10) for 5 months. ²² No losses were observed in any of the batches compared to the initial counts in any conditions.

Three batches of Bovacillus™ 10 were mixed into a standard dairy cow vitamin‐mineral premixture at an intended inclusion rate of 1.0 × 10¹¹ CFU/kg feed and were kept for 3 months at ambient temperature. ²³ The results showed negligible losses in all batches compared to the initial counts.

Three batches of Bovacillus™ 10 (three replicates per batch) were individually mixed into a standard dairy cow feed (based on wheat, barley, maize, rapeseed and soybean meal) in mash form at an intended inclusion rate of 8.6 × 10⁸ CFU/kg feed. The feed was pelleted at 75, 85 and 95°C for 30 s. ²⁴ The results showed no losses (< 0.5 log CFU/kg) during pelleting in all batches compared to the initial counts. The stability of the mash and pelleted feeds was investigated during storage for up to 3 months at ambient temperature (ca. 21°C). ²⁵ The results showed no losses (< 0.5 log CFU/g).

The stability of Bovacillus™ WS (three batches) in water (intended inclusion rate of 1.1 × 10⁸ CFU/L) was studied during storage at ambient temperature (25°C) for 48 h. The results showed no losses (< 0.5 log CFU/g) in any of the batches tested. ²⁶

Ten sub‐samples were taken from the feed pelleted at 85°C and analysed for total bacilli counts to evaluate the homogeneity distribution of the additive in the feed. Based on the 10 samples, the coefficient of variation was 15%. ²⁷

3.1.4. Conditions of use

The additive is intended for use in feed and water for dairy cows and other dairy ruminants at a minimum inclusion level of 3.8 × 10⁸ CFU/kg complete feed or 7.4 × 10⁷ CFU/L water for drinking. The applicant suggested a daily dose of 9.6 × 10⁹ CFU per head.

Bovacillus™ 10 is recommended for use in feed, and Bovacillus™ WS is recommended for use in water.

3.2. Safety

3.2.1. Safety for the target species, consumers and environment

The additive consists of two bacterial strains, which are considered by EFSA to be eligible for the qualified presumption of safety (QPS) approach (EFSA BIOHAZ Panel, 2023). This approach requires the identity of the strains to be conclusively established and evidence that they do not show acquired resistance to antibiotics of human and veterinary importance or toxigenic potential. Moreover, the B. paralicheniformis strain should also lack the ability to produce bacitracin. In view of the FEEDAP Panel, the identity of the strains B. paralicheniformis and B. subtilis was established, and all the qualifications were met. Consequently, B. paralicheniformis DSM 33902 and B. subtilis DSM 33903 are presumed safe for the target species, consumers and the environment. In addition, since no safety concerns are expected from other components of the additive, the additive is also considered safe for the target species, consumers and the environment.

3.2.2. Safety for the user

The dusting potential of the two formulations of the additive (Section 3.1.2) indicates that exposure by inhalation is likely.

The additive contains microorganisms and, therefore, should be considered a skin and respiratory sensitiser.

The skin irritation potential of Bovacillus™ 10 was tested in a study performed according to the OECD Testing Guideline (TG) 404, which showed that the additive is not a skin irritant (UN GHS ‘No Category’). ²⁸

The eye irritation potential of Bovacillus™ 10 was tested in a study performed according to the OECD TG 438, which showed that the additive is not an eye irritant (UN GHS ‘No Category’). ²⁹

The applicant did not submit any data on the irritation potential of Bovacillus™ WS. Bovacillus™ WS contains maltodextrin as carrier while Bovacillus™ 10 includes calcium carbonate, both included at similar levels. The Panel has previously evaluated the skin and eye irritation potential of another product of the same company, consisting of a preparation containing viable cells of another strain (Pediococcus pentosaceus DSM 23688) and maltodextrin as carrier (EFSA FEEDAP Panel, 2024a, 2024b). The results of these tests showed no skin nor eye irritation potential. Therefore, considering the results of these tests together with the ones submitted in the current application for Bovacillus™ 10, in which the irritation potential of the Bovacillus™ strains was evaluated, the Panel concludes that Bovacillus™ WS is not irritant to skin and eyes.

3.2.2.1. Conclusions on safety for the user

Bovacillus™ 10 is not a skin or eye irritant but is considered a skin and respiratory sensitiser. Dermal and inhalation exposure is considered a risk. This conclusion can be extended to the other formulation of the additive, Bovacillus™ WS.

3.3. Efficacy

3.3.1. Efficacy for dairy cows

The applicant provided three long‐term trials with lactating dairy cows aiming at assessing the effect of Bovacillus™ on the zootechnical performance at the proposed conditions of use. The details of the study design are provided in Table 1, and the main results are in Table 2.

TABLE 1.

Trial design and use level of the efficacy trials performed in dairy cows.

Trial	Total N (animals/replicate) replicae/ group	Breed DIM (duration) parity	Feeding method (composition)	Additive supplementation	Groups (CFU/kg complete feed)
					Intended	Calculated*
1	76 (1) 38	Holstein Friesian 27 ± 4 (13 weeks) 2–4	Total mixed ration ■■■■■	Top‐dressing (with molasses)	0 3.8 × 108	– 3.25 × 108
2	68 (1) 34	Holstein Friesian × Swedish Red 50 ± 6 (12 weeks) 1–5	Partial mixed ration ■■■■■ plus concentrate ■■■■■	Complementary feed (concentrate)	0 3.8 × 108	– 3.08 × 108
3	76 (1) 38	Holstein Friesian 42 ± 6 (15 weeks) 1–6	Partial mixed ration ■■■■■ plus concentrate ■■■■■	Complementary feed (concentrate)	0 3.8 × 108	– 3.41 × 108

Note: Total N: total number of animals; DIM: days in milk.

^*The counts of the total bacilli were analysed in the premix used for top‐dressing the feed in trial 1 (1.8 × 10⁹ CFU/kg) or in the corresponding complementary feeds in trials 2 (9.5 × 10⁹ CFU/kg) and 3 (1.5 × 10¹⁰ CFU/kg). The bacilli counts in feed (mg/kg) were calculated based on these analytical data and the dry matter intake of the animals during the experiment.

TABLE 2.

Effect of Bovacillus™ on the production performance of dairy cows.

Trial
Groups (CFU/kg feed)	Dry matter intake (kg/day)	Final body weight (kg)	Milk yield (kg)	Feed efficiency	ECM 1 efficiency	Milk protein (%)	Milk fat (%)
1	0	24.2	715	39.6	1.67	1.67b	2.84	3.65
	3.8 × 108	24.1	723	40.3	1.69	1.72a	2.86	3.82
2	0	27.3	690	40.1b	1.48b	1.54	3.59	4.29
	3.8 × 108	27.3	683	41.3a	1.52a	1.58	3.56	4.21
3	0	25.0	709	38.0	1.54b	1.56	3.30	3.54
	3.8 × 108	24.5	708	38.3	1.59a	1.59	3.32	3.46

¹ Energy‐corrected milk yield. a,b: Mean values within a trial and within a column with a different superscript are significantly different (P<0.10)

In all trials, cows were ■■■■■ randomly allocated to two groups: one receiving a non‐supplemented diet (control) and another one receiving a Bovacillus™‐supplemented diet.

In trial 1, all cows were housed in the same collective barn ■■■■■ for individual feed intake and fed ■■■■■ with a total mixed ration (TMR) for ad libitum intake ■■■■■ Each day■■■■■ the feed was top‐dressed with molasses either not supplemented (control) or supplemented with ■■■■■ Bovacillus™ 10/head per day (equivalent to an intended level of 3.8 × 10⁸ CFU/kg complete feed). The concentration of the additive was analysed in the batch of the test item used in the trial, ³⁰ and the extrapolation to the level in complete feed was estimated based on the average dry matter intake (DMI).

In trials 2 and 3, cows were housed in collective barns ■■■■■ allowing for individual feed intake monitoring. The cows received a partial mixed ration ad libitum complemented with a protein concentrate offered separately. The concentrate was either not supplemented (control) or supplemented with ■■■■■ Bovacillus™ 10/head per day (equivalent to an intended level of 3.8 × 10⁸ CFU/kg complete feed). The concentration of the additive was analysed in the protein concentrate, ³¹ and the extrapolation to the level in the complete feed was estimated based on the average DMI.

In all cases, it was reported that all the cows consumed the supplement, and no leftovers were detected.

Mortality and health status were monitored throughout the experiment. The cows were weighed at the start and end of the experimental period, and the individual feed intake and milk yield were ■■■■■ recorded. ■■■■■ milk from each cow was sampled to determine total solids, fat, protein, lactose, urea and somatic cell counts. The ■■■■■ dry matter intake, feed efficiency (kg of milk/kg of dry matter consumed) and energy‐corrected feed efficiency ³² were calculated per cow for the whole experimental period.

In all trials, the data for performance parameters and milk composition were analysed ■■■■■ The significance level was set at 0.10.

The inclusion of Bovacillus™ in the feed of dairy lactating cows at the proposed use level showed an improvement in the milk production performance in all three trials. In trial 1, the supplemented groups showed higher fat‐corrected milk yield (FCM) ³³ (40.3 vs. 42.2 kg/day) and better ECM and FCM ³⁴ (1.70 vs. 1.77) efficiency compared with the control. In trials 2 and 3, the supplemented groups showed better milk feed efficiency compared to the control , and, in the case of trial 2, a higher milk yield was also observed. No effect on any of the milk quality parameters measured was observed.

3.3.2. Conclusions on efficacy

Based on the results of the efficacy trials submitted, the FEEDAP Panel concludes that Bovacillus™ has the potential to be efficacious as a zootechnical additive for dairy cows at 3.8 × 10⁸ CFU/kg complete feed. The FEEDAP Panel considers that for ruminants, a fixed ratio of feed‐to‐water intake cannot be set to define a minimum use level in water. However, considering that the daily dose of the additive suggested by the applicant was effectively tested in all trials (9.6 × 10⁹ CFU per cow), the use level in water could be calculated based on the daily water intake. This conclusion can be extrapolated to other dairy ruminants.

3.4. 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 ³⁵ and Good Manufacturing Practice.

4. CONCLUSIONS

Bovacillus™ is considered safe for the target species, consumers and the environment under the proposed conditions of use.

Bovacillus™ 10 and WS are considered non‐irritants to the skin and eyes, but skin and respiratory sensitisers, and any exposure through the skin and respiratory tract are considered a risk.

Bovacillus™ has the potential to be efficacious as a zootechnical additive in dairy cows and other dairy ruminants when used in feed and water. The minimum use level in feed could be established at 3.8 × 10⁸ CFU/kg complete feed. The minimum use level in water can be established based on the daily dose per animal (9.6 × 10⁹ CFU per head) and the daily water intake.

ABBREVIATIONS

AMR

antimicrobial resistance

ANI

average nucleotide identity

CFU

colony forming unit

ECM

energy‐corrected milk yield

DIM

days in milk

DMI

dry matter intake

DSM

Leibniz Institute DSMZ ‐ German Collection of Microorganisms and Cell Cultures GmbH

EURL

European Union Reference Laboratory

FCM

fat‐corrected milk yield

FEEDAP

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

LOQ

limit of quantification

MIC

minimum inhibitory concentration

OECD

Organisation for Economic Co‐operation and Development

QPS

qualified presumption of safety

TG

Testing guideline

TMR

total mixed ration

UN GHS

Globally Harmonized System of Classification and Labelling of Chemicals

WGS

whole genome sequence data

REQUESTOR

European Commission

QUESTION NUMBER

EFSA‐Q‐2023‐00454

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.

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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.

EFSA FEEDAP Panel (EFSA Panel on Additives and Products or Substances used in Animal Feed) , Villa, R. E. , Azimonti, G. , Bonos, E. , Christensen, H. , Durjava, M. , Dusemund, B. , Gehring, R. , Glandorf, B. , Kouba, M. , López‐Alonso, M. , Marcon, F. , Nebbia, C. , Pechová, A. , Prieto‐Maradona, M. , Röhe, I. , Theodoridou, K. , López‐Puente, S. , Anguita, M. , … Ortuño, J. (2025). Safety and efficacy of a feed additive consisting of Bacillus paralicheniformis DSM 33902 and Bacillus subtilis DSM 33903 (Bovacillus™) for dairy cows and other dairy ruminants (Chr. Hansen A/S). EFSA Journal, 23(4), e9426. 10.2903/j.efsa.2025.9426