Bifidobacterium longum CNCM I-5642 (4e001) for dogs: counts by pour plate on reinforced clostridial agar (LOQ 100 CFU/g), identification by PFGE — 3.5 × 10⁹ CFU/kg feed, at least 6 weeks recommended, efficacy only in the second EFSA opinion
Commission Implementing Regulation (EU) 2025/1776, published on 9 September 2025, authorises until 29 September 2035 a preparation of Bifidobacterium longum CNCM I-5642 (PP102I, holder: Nestlé Enterprises S.A. – Division Nestlé Purina Petcare Europe) as a physiological condition stabiliser for dogs — improving stress resilience in realistic situations. In its opinion of 29 June 2022 EFSA could not confirm efficacy (the application for cats was withdrawn); only a new cross-over study in 20 dogs, assessed on 28 January 2025, was enough for a positive conclusion. The additive is an eye irritant and a skin and respiratory sensitiser.
- Methods in the annex: identification — DNA sequencing or PFGE (CEN/TS 17697); count in the additive and compound feed — pour plate method on reinforced clostridial agar (the PL text describes it as an “enriched agar medium for the detection, isolation and enumeration of Clostridium”). According to the EURL report the method was single-laboratory validated: at 1.3 × 10¹¹ CFU/g in the additive Sr and Sip = 0.06 log₁₀, in feed (9.2 × 10⁸–8.0 × 10⁹ CFU/g) Sr 0.04–0.13 and Sip 0.04–0.17 log₁₀; LOQ 100 CFU/g feed according to ISO 7218.
- Additive (2022 opinion): min. 5 × 10¹⁰ CFU/g; five batches on average 7.72 × 10¹⁰ CFU/g (6.00–9.50 × 10¹⁰). Cadmium 0.061 mg/kg, lead 0.018 mg/kg, mercury 0.016 mg/kg, arsenic below the LOQ in two batches and 0.017 mg/kg in the third; manganese 1.9–2.7, zinc 9.1–9.8 mg/kg; PCDD/Fs 0.222 ng WHO-TEQ/kg (0.099–0.463); mycotoxins below the LOD. Dust-free (0 mg/m³, Stauber-Heubach); no losses after 8 months at 4 °C and in a dry complementary feed after 88 days at 23 °C and 48 days at 32 °C; homogeneity in the feed — CV 15 % (36 subsamples).
- New study (2025 opinion): 20 adult dogs, cross-over design — 2 × 42 days with a 23-day washout, 1 × 10⁹ CFU/dog per day in 1 g of complementary feed. Challenge: clinic visit, 10-minute car ride, second visit, 30 minutes of recovery. At the first visit tympanic temperature 38.1 versus 38.4 °C, LF/HF 0.22 versus 0.33; in the car standing 13.9 versus 28.9 s, lying 345 versus 278 s; during recovery RMSSD 102 versus 83 ms, pNN50 34.4 versus 23.5 %. Serum cortisol was unchanged; yawning increased (0.42 versus 0.23). The significance level was set at 0.10.
- Conditions in the annex: min. 3.5 × 10⁹ CFU/kg complete feed (equivalent to 1 × 10⁹ CFU per dog per day); on the label of the additive, premixtures and compound feed: “It is recommended to use for at least 6 weeks”; state storage conditions and stability to heat treatment. The OECD 492 test reduced cell viability to 33 % — the additive is an eye irritant (no category can be assigned), not a skin irritant (OECD 439); skin, eye and respiratory protection.