1. Introduction

The FSA and FSS have undertaken an assessment for a feed additive (PP102I) consisting of Bifidobacterium longum CNCM I-5642, (Nestlé Enterprises S.A., Division Nestlé Purina Petcare Europe, Entre-Deux-Villes 12, 1800 Vevey, Switzerland) under Assimilated Regulation (EC) No 1831/2003 (EC, 2003) under the category of ‘zootechnical additives’ and functional group ‘physiological condition stabiliser’. The additive is proposed to be used at an inclusion rate of 1 x 109 CFU/dog daily.

To support the safety assessment by FSA and FSS, the ACAF provided advice to the FSA and FSS outlined in this document.

In line with Article 8 of 1831/2003, the assessment has considered whether the feed additive complies with the conditions laid down in Article 5, including: safety considerations for human, animal and environmental health; efficacy of the additive for its intended effect; potential impairment of the distinctive features of animal products. This, and the guidance put in place by EFSA for the evaluation of feed additive applications, has formed the basis and structure for the assessment.

The dossier was evaluated by the by ACAF at their July 2023 meeting, after which a request for further information was communicated to the applicant. The applicant provided responses to the ACAFs requests in October 2023 and January 2024.

The views of ACAF have been taken into account in this safety assessment which represents the opinion of the FSA/FSS.

Table 1.Table showing products included in this assessment
Title Product type Intended use/s Intended species or categories of animals
Bifidobacterium longum CNCM I-5642 Feed additive Zootechnical additive Dogs

2. Assessment

2.1. Section II: Identity, characterisation and conditions of use

The additive’s active substance Bifidobacterium longum (CNCM I-5642) has a minimum guaranteed activity of 5 x 1010 CFU/g. The applicant provided data from several batches supporting the identification values outlined below (Table 2):

Table 2.Identity table Bifidobacterium longum CNCM I-5642
Composition
Bifidobacterium longum CNCM I-5642 7.72 x 1010 CFU/g
Appearance
Dry powder
Chemical-physical specifications
Bulk density 0.439 – 0.493 g/ml
Dusting potential 0 mg/m3
Particle size <100 µm –30%
<50 µm – 9%
<10 µm – 1.3%
Impurities
Bacillus cereus <10 CFU/g
Coliforms <10 CFU/g
Salmonella Absent in 25g
E. coli Absent in 25g
Enterobacteriaceae Absent
Moulds <10 CFU/g
Yeasts <10 CFU/g
Arsenic <0.017 mg/kg
Lead 0.017 – 0.019 mg/kg
Cadmium 0.058 – 0.067 mg/kg
Chromium 0.085 – 0.103 mg/kg
Sum of dioxins (WHO-PCDD/F-TEQ) 0.099 – 0.463 TEQ (pg/g)
Sum of dioxins and dl-PCBs (WHO-PCDD/F-TEQ) 0.112 – 0.505 TEQ (pg/g)

The Committee reviewed the genetic characterisation of the active ingredient and noted that a comprehensive analysis had been performed confirming the identity of the active substance as B. longum. Through this characterisation the applicant demonstrated that the additive meets the requirements for the qualified presumption of safety (QPS) approach to assessment. The Committee noted inconsistencies in the conditions of use of the additive and the proposed method of administration of the additive. Following a request for information, the applicant provided further material that allowed a comprehensive assessment to be performed. The additive was demonstrated to be stable for 8 months at 4°C. In feed, the additive was shown to be stable for 48 weeks at 32°C at 70% humidity, and for 88 weeks at 23°C at 60% humidity. Homogeneity in feed was also demonstrated.

The proposed conditions of use of the additive are described in Table 3:

Table 3.Proposed conditions of use of Bifidobacterium longum CNCM I-5642
Proposed mode of use in animal nutrition
Additive Bifidobacterium longum (CNCM I-5642)
Registration number/ EC No/No 4exx
Category(-ies) of additive 4. Zootechnical feed additive
Functional group(s) of additive e. Physiological condition stabiliser
Description
Description Chemical formula Purity criteria Method of analysis
Preparation of live B. longum (CNCM I-5642) Powder form with a min. 5 x 1010 CFU/g Complies with EU feed hygiene regulation LI 00.794
Trade name (if appropriate) PP102I
Name of the holder of authorisation (if appropriate) Nestle Enterprises S.A (NESA)
Proposed conditions of use
Species or category of animal Min-max Age Min. content Max. content Withdrawal period
 
Dogs -- 1 x 109 CFU/dog daily -- --

2.1.1. Conclusions on Section II

The ACAF concluded the additive was correctly characterised and is stable for 8 months at 4°C. In feed, the additive is stable for 48 weeks at 32 °C (70% humidity), and for 88 weeks at 23°C (60% humidity). Homogeneity in feed was also demonstrated.

No further concerns were raised for Section II of these dossiers.

2.2. Section III: Safety

2.2.1. Safety for the target species

The applicant demonstrated the species B. longum to meet the requirements for the qualified presumption of safety (QPS) approach to assessment, therefore the additive is presumed safe for the target species.

2.2.2. Safety for the consumer

The applicant demonstrated the species B. longum to meet the requirements for the QPS approach to assessment, therefore the additive is presumed safe for the consumer.

2.2.3. Safety for the user

The applicant presented the following tests to evaluate the safety of the additive for the user/worker:

  • In vitro skin irritation test (EPISKIN reconstructed human epidermis model – OECD 439)

  • In vitro eye irritation test (reconstructed human cornea-like epithelium model – OECD 492)

From the data provided the Committee concluded that the additive is not an irritant to the skin and is an irritant to the eyes. Owing to the absence of data the additive should be considered a skin sensitiser. As a microorganism product the additive should be regarded as a respiratory sensitiser. It was noted that due to the presence of particles in the smaller particle size distribution fraction, exposure through inhalation should be avoided.

2.2.4. Safety for the environment

The applicant demonstrated the species B. longum to meet the requirements for the QPS approach to assessment, therefore the additive is presumed safe for the environment.

2.2.5. Conclusions on safety

As the species B. longum was demonstrated to meet the criteria for the QPS approach to assessment, the ACAF concluded that the additive can be considered safe for the consumer, the target species, and the environment.

The ACAF concluded that the additive is not an irritant to the skin and is an irritant to the eyes. The additive should be considered a potential skin sensitiser and respiratory sensitiser.

2.3. Section IV: Efficacy

The applicant provided three long-term efficacy studies to support the efficacy claims in dogs. An overview of the trials is presented in Table 4:

Table 4.Overview of efficacy trials in dogs
Study Replicates/Treatment Target B. longum Analysed B. longum Duration
1 24 dogs; 2 treatments. 24 replicates per treatment (cross-over) 1 x 109 CFU/dog/day 2.35 x 108 CFU/dog/day 17 weeks (7+3+7)
2 20 dogs; 2 treatments. 20 replicates per treatment (cross-over) 1 x 109 CFU/dog/day 9.5 x 108 CFU/dog/day 15 weeks (6+3+6)
3 20 dogs; 2 treatments. 20 replicates per treatment (cross-over) 1 x 109 CFU/dog/day 9.83 x 108 CFU/dog/day 15 weeks (6+3+6)

The Committee reviewed each of the studies and the physiological and behavioural parameters investigated, noting the trials were performed to a high standard. The three studies performed in dogs demonstrated that the animals treated with B. longum were less reactive, calmer and in a better affective state when subjected to routine stressors.

The ACAF concluded that B. longum is efficacious in dogs when included in feed at the proposed dose of 1 x 109 CFU/dog daily.

2.3.1. Conclusions on efficacy

Based on the information presented in three efficacy trials, the ACAF concluded that Bifidobacterium longum CNCM I-5642 is efficacious in dogs when included in feed at the proposed dose of 1 x 109 CFU/dog daily.

3. Analytical methods evaluation

Conclusions on the analytical methods are presented here as an extract from the Evaluation Report of the European Union Reference Laboratory for Feed Additives on the Method(s) of the Analysis for Bifidobacterium longum (CNCM I-5642) (EURL-FA, 2021):

“For the identification of Bifidobacterium longum (CNCM I-5642), the EURL recommends for the official control Pulsed Field Gel Electrophoresis (PFGE), a generally recognised methodology for the genetic identification of bacterial strains. For the enumeration of Bifidobacterium longum (CNCM I-5642) in the feed additive and feedingstuffs, the Applicant submitted a single-laboratory validated and further verified pour plate count method using a reinforced clostridial agar medium. The following performance characteristics were obtained in the frame of the validation and verification studies: (i) for the average measured content of the active substance in the feed additive of 1.3 x 1011 CFU/g: a standard deviation for repeatability (Sr) and intermediate precision (Sip) of 0.06 log10 CFU/g; (ii) for the average measured content of the active substance in feedingstuffs ranging from 9.2 x 108 to 8.0 x 109 CFU/g: Sr ranging from 0.04 to 0.13 log10 CFU/g and Sip ranging from 0.04 to 0.17 log10 CFU/g. Furthermore, a limit of quantification (LOQ) of 100 CFU/g feedingstuffs can be derived following the recommendations of ISO 7218 standard. Based on the performance characteristics and the experimental data available, the EURL recommends the pour plate count method using a reinforced clostridial agar medium for the official control for the enumeration of Bifidobacterium longum (CNCM I-5642) in the feed additive and feedingstuffs.”

FSA/FSS accepts the EURL analytical method evaluation reports. FSA/FSS determined the analytical method as appropriate for official controls for this feed additive.

4. Conclusions

The ACAF concluded the additive was correctly characterised and is stable for 8 months at 4°C. In feed, the additive is stable for 48 weeks at 32°C (70% humidity), and for 88 weeks at 23°C (60% humidity). Homogeneity in feed was also demonstrated.

As the species B. longum was demonstrated to meet the criteria for the QPS approach to assessment, the ACAF concluded that the additive can be considered safe for the consumer, the target species, and the environment.

The ACAF concluded that the additive is not a skin irritant. The additive is an eye irritant and should be considered a potential skin sensitiser and respiratory sensitiser.

The ACAF concluded that Bifidobacterium longum CNCM I-5642 is efficacious when included in feed at the proposed dose of 1 x 109 CFU/dog daily.

The FSA/FSS agree with the conclusions reached by the ACAF. FSA/FSS accepts the EURL analytical method evaluation reports. FSA/FSS determined the analytical method as appropriate for official controls for this feed additive.

Abbreviations

Acronym Definition
ACAF    Advisory Committee on Animal Feedingstuffs   
CFU Colony forming units
EFSA European Food Safety Authority
EU European Union
FSA Food Standards Agency
FSS Food Standards Scotland
QPS Qualified presumption of safety
LOQ Limit of quantification
PFGE Pulsed Field Gel Electrophoresis
Sr Standard deviation for repeatability
Sip Intermediate precision

Acknowledgements

With thanks to the members of the ACAF during the course of the assessment, who were: Professor Nicholas Jonsson, Martin Briggs, Professor Emily Burton, Professor Katrina Campbell, Professor Matthew Fisher, Hannah Kane, Susan MacDonald, Dr. Oonagh Markey, Christine McAlinden, Dr. Donald Morrison, Derek Renshaw, Dr. Michael Salter, Dr. Adam Smith, Dr. Helen Warren and Dr. Nick Wheelhouse. Hannah Kane declared an indirect conflict of interest and remained in the meetings for discussion.