1. Charakterystyka żywności / składnika

Introduction on the process used for characterisation of food constituents that are microorganisms: Microorganisms or microbes (e.g. bacteria) are living organisms, and can change over time depending on culture conditions. Correct identification of the microorganism‟s species and strain is of critical importance, as the observed effects are species and strain specific. The appropriate classification, identification and nomenclature of microorganisms constitute the starting point for the assessment of microbial properties. Classification assigns an organism to a known taxonomic group according to its similarity to that group. This allows the prediction of the properties of the microorganism on the basis of what is already known about the taxa. A reliable identification confirms the identity of the strain(s) used in a given process and requires the use of appropriate methods. Traditional phenotypic identification of bacteria is not always reliable since certain species cannot be distinguished by these methods. Molecular techniques have emerged in recent years as a replacement or complement to traditional phenotypic tests. DNA-DNA hybridization has become the generally accepted standard for determination of bacterial species identification. However this technique is difficult to perform and requires an expertise not normally present in the food industry. For these reasons phylogenetically based approaches such as sequence analysis of the 16S rRNA gene has proven to be a useful tool for bacterial identification. The EU-funded PROSAFE project concluded that biochemical tests should not be used as a stand-alone approach for identification of bacterial cultures (Vankerckhoven et al., 2008). The use of 16S rRNA gene sequence analysis was considered the best tool for routine species identification. Moreover, the use of sequence-based methods, such as 16S rRNA gene sequencing, was encouraged given their high reproducibility and data exchangeability (Vankerckhoven et al., 2008). The FAO/WHO expert group (FAO, 2006) recommends that phenotypic tests should be done first, followed by genetic identification, using methods such as DNA-DNA hybridization or 16S rRNA sequence analysis. Nevertheless, it is important to underline that in some cases 16S rRNA sequencing has a limited resolution and it may not be enough for discrimination of closely related species (Felis and Dellaglio, 2007; Vankerckhoven et al., 2008) being necessary to use other methods.
For the strain identification (characterisation of the strain by genetic typing), the FAO/WHO working group also recommended that strain typing has to be performed with a reproducible genetic method or using a unique phenotypic trait (FAO, 2006). DNA macrorestriction followed by Pulsed Field Gel Electrophoresis (PFGE) is considered as the generally accepted standard (FAO, 2006) and it has been extensively used for differentiating commercial microorganism strains. Other discriminatory molecular methods, such as Randomly Amplified Polymorphic DNA (RAPD) or Amplified rDNA restriction analysis (ARDRA) among others, are also available for strain characterisation. Hence, species identification and sufficient characterisation (genetic typing) at strain level by using internationally accepted molecular methods is needed. In addition, strains should be named according to the International Code of Nomenclature. Although there is no direct requirement on deposition of the particular strain in an internationally recognised culture collection, the FAO/WHO (FAO, 2006) recommends that strains should also be deposited in an internationally recognised culture collection (with access number). These will assure the tracking and access of scientists and regulatory authorities to the strain and related information in case it is needed. In the context of the Regulation (EC) nº 1924/2006, the purposes of characterisation are to confirm the identity of the food/constituent that is the subject of the health claim, and to establish that the studies provided for substantiation of the health claim were performed with the food/constituent in respect of which the health claim is made. Although not required for substantiation of a claim, characterisation should also be sufficient to allow control authorities to verify that the food/constituent which bears a health claim is the same one that was the subject of a community authorisation. The Panel has decided to use the following criteria for characterisation of food constituents that are microorganisms, which are the subject of health claims:
 Species identification by DNA-DNA hybridization or 16S rRNA sequence analysis.
 Strain identification by DNA macrorestriction followed by PFGE, RAPD, ARDRA or other internationally accepted genetic typing molecular methods.
Only when these two criteria were fulfilled, the microorganism was considered to be sufficiently characterised. In case of combination of several microorganisms, the Panel considers that if one microorganism used in the combination is not sufficiently characterised, the combination proposed is not sufficiently characterised. The characterisation of food constituents that are microorganisms, which are the subject of health claims pursuant to Article 13 of the Regulation (EC) nº 1924/2006, is based on evaluation of available references up to 31 December 2008, including the following:
 The information provided by the Member States in the consolidated list of Article 13 health claims and references that EFSA has received from Member States or directly from stakeholders;
 Generally available data obtained by searching PubMed and Web of Science databases by using the strain name as search term.

1.39. Characterisation of a combination of “Propionibacterium freudenreichii SI 41 and Propionibacterium freudenreichii SI 26” (ID 941)

The food constituent that is the subject of the health claim is a combination of “Propionibacterium freudenreichii SI 41 and Propionibacterium freudenreichii SI 26”. Propionibacterium freudenreichii SI 41 - The strain Propionibacterium freudenreichii SI 41 species identification/characterisation is indicated in the references provided (Jan et. al., 2002; Jan et al., 2000). In addition the strain has been further characterised in different published reports on stress response or transcarboxylase gen characterisation (Jan et al., 2000; Herve et al., 2007). The Panel considers that Propionibacterium freudenreichii SI 41, which is the subject of the health claim ID 941, is sufficiently characterised. No indication of the deposit of the strain in an internationally recognised culture collection was found in the references provided or the literature.
Propionibacterium freudenreichii SI 26 - The strain Propionibacterium freudenreichii SI 26 species identification/characterisation is not indicated in the references provided and no information was found in the literature. The Panel considers that Propionibacterium freudenreichii SI 26, which is the subject of the health claim ID 941, is not sufficiently characterised.
No indication of the deposit of the strain in an internationally recognised culture collection was found in the information provided or the literature. The Panel considers that the combination of “Propionibacterium freudenreichii SI 41 and Propionibacterium freudenreichii SI 26”, which is the subject of the health claim ID 941, is not sufficiently characterised.

2. Znaczenie oświadczenia dla zdrowia człowieka

The claimed effect, which is proposed for further assessment, is “beneficially affects the intestinal flora by increasing bifidobacteria”. The proposed target population is the general population.
The Panel notes that increasing the number of any groups of microorganisms, including lactobacilli and/or bifidobacteria, is not considered to be a beneficial physiological effect in itself, but needs to be linked to a beneficial physiological effect.

3. Naukowe uzasadnienia wpływu na zdrowie człowieka - Zwiększenie ilości bakterii jelitowych

The references cited in relation to this claim included one human study, which addressed the effects of P. freudenreichii SI 41 and P. freudenreichii SI 26 on faecal propionibacteria and bifidobacteria concentrations, and on segmental colonic transit time (Bouglé et al., 1999), and two human studies on the effect of P. freudenreichii SI41 alone (and not the combination of the strains that is the subject of the claim) in different matrices (e.g. fermented milk, and classical or acid-resistant capsules) on, for example, propionibacterium populations in faeces, survival capacity/viability of the strain, or the metabolic activity of the strain in the gut (Hervé et al., 2007; Jan et al., 2002). The Panel considers that the evidence provided does not establish that increasing numbers of gastro- intestinal microorganisms is a beneficial physiological effect.
The Panel concludes that a cause and effect relationship has not been established between the consumption of a combination of Propionibacterium freudenreichii SI 41 and Propionibacterium freudenreichii SI 26 and a beneficial physiological effect related to increasing numbers of gastro- intestinal microorganisms.

Oświadczenia zdrowotne

ID 941 - Propionibacterium freudenreichii SI 41, Propionibacterium freudenreichii SI 26

1. Charakterystyka żywności / składnika

1.39. Characterisation of a combination of “Propionibacterium freudenreichii SI 41 and Propionibacterium freudenreichii SI 26” (ID 941)

2. Znaczenie oświadczenia dla zdrowia człowieka

3. Naukowe uzasadnienia wpływu na zdrowie człowieka - Zwiększenie ilości bakterii jelitowych

Warunki i możliwe ograniczenia stosowania oświadczenia