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Bacillus subtilis KCCM 10673P, Aspergillus oryzae KCTC 10258BP -1o01

Viable cells of Bacillus subtilis KCCM 10673P and Aspergillus oryzae KCTC 10258BP

Details

Publication date
23 April 2010 (Last updated on: 6 June 2016)

Description

In the current application authorisation is sought for the probiotic PepSoyGen-C, which consists of two active agents Bacillus subtilis GR-101 and Aspergillus oryzae GB-107, under the category 'zootechnical additives', functional group 'other zootechnical additives' according to Annex I of Regulation (EC) No 1831/2003. Specifically, authorization is sought for the use of Bacillus subtilis GR-101 and Aspergillus oryzae GB-107 for piglets, chicken for fattening, calves for fattening and for rearing, fish (salmonidae and other fish), lambs and goats for rearing and fattening and for dogs. The product is to be used to treat feed materials to diminish anti-nutritional factors and will enter animals in live form. The proposed conditions of use do not include minimum or maximum concentrations of the feed additive. According to the applicant the product as such will not be placed on the market, but feed materials previously treated with the product.

For the enumeration of Bacillus subtilis GR-101 in premixtures and feedingstuffs the CEN method (EN 15784) has been validated at a range between 105 and 109 CFU/g. The performance characteristics of the CEN method reported after logarithmic transformation of measured values (CFU) are - for the premixtures: (1) a standard deviation for repeatability (sr) of 0.09 log10 CFU/g and (2) a standard deviation for reproducibility (sR) of 0.32 log10 CFU/g; - for the feedingstuffs: (1) s = 0.07 log10 CFU/g and (2) sR = 0.35 log10 CFU/g and a limit of detection (LOD) of 1x105 CFU/kg in feedingstuffs.

However, since this specific application does not include target levels of Bacillus subtilis GR-101 in feedingstuffs, the CRL cannot evaluate the suitability of this CEN standard.

For the enumeration of Aspergillus oryzae GB-107 the applicant proposes internationally recognised US FDA/CFSAN BAM spread plate method for enumeration of yeasts, moulds and mycotoxins. The applicant considers that further validation or verification is not necessary since this is an official US method. However, another international standard exists for the enumeration of yeasts and moulds (ISO 21527–1). No performance characteristics of this spread plate method were provided except the LOD of 1x105 CFU/kg in feed. As this method was not tested on the product, the CRL cannot evaluate the suitability of this ISO method for official control. Molecular methods were used by the applicant for identification of active agents. The CRL recommends for official control for Bacillus subtilis GR-101, Pulsed Field Gel Electrophoresis (PFGE), a generally recognised standard methodology for bacterial identification. For Aspergillus oryzae GB-107 the CRL recommends for official control Polymerase Chain Reaction (PCR), generally recognised standard methodology for identification of yeasts and moulds.

Further testing or validation is not considered necessary.

Recommended Method

as of:

- Commission Implementing Regulation (EU) 2020/180

Enumeration of Bacillus subtilis KCCM 10673P in the feed additive, premixtures and feedingstuffs: 

- Spread plate method on tryptone soya agar (EN 15784)

Identification of Bacillus subtilis KCCM 10673P in the feed additive: 

- Pulsed Field Gel Electrophoresis (PFGE)

Identification of Aspergillus oryzae KCTC 10258BP in the feed additive: 

- Polymerase Chain Reaction (PCR) typing

Files

  • 2 FEBRUARY 2022
FinRep-FAD-2009-0007.pdf