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.

1.33. Characterisation of a combination of “Lactobacillus gasseri PA 16/8, Bifidobacterium bifidum MF 20/5, Bifidobacterium longum SP 07/3” (ID 931, 933)

The food constituent that is the subject of the health claim is a combination of “Lactobacillus gasseri PA 16/8, Bifidobacterium bifidum MF 20/5, Bifidobacterium longum SP 07/3”. Lactobacillus gasseri PA 16/8 - No reference to strain Lactobacillus gasseri PA 16/8 species identification/characterisation is indicated in the references provided and no information regarding the strain identification/characterisation was found in the literature. The Panel considers that Lactobacillus gasseri PA 16/8, which is the subject of the health claims ID 931, 933, is not sufficiently characterised.
No indication of the deposit of the strain in an internationally recognised culture collection was reported in the information provided or the literature.
Bifidobacterium bifidum MF 20/5 - No reference to strain Bifidobacterium bifidum MF 20/5 species identification/characterisation is indicated in the references provided and no information regarding the strain identification/characterisation was found in the literature. The Panel considers that Bifidobacterium bifidum MF 20/5, which is the subject of the health claims ID 931, 933, is not sufficiently characterised. No indication of the deposit of the strain in an internationally recognised culture collection was reported in the information provided or the literature. Bifidobacterium longum SP 07/3 - No reference to strain Bifidobacterium longum SP 07/3 species identification/characterisation is indicated in the references provided and no information regarding the strain identification/characterisation was found in the literature. The Panel considers that Bifidobacterium longum SP 07/3, which is the subject of the health claims ID 931, 933, is not sufficiently characterised. No indication of the deposit of the strain in an internationally recognised culture collection was reported in the references provided or the literature. The Panel considers that the combination of “Lactobacillus gasseri PA 16/8, Bifidobacterium bifidum MF 20/5, Bifidobacterium longum SP 07/3”, which is the subject of the health claims ID 931, 933, is not sufficiently characterised.

2. Znaczenie oświadczenia dla zdrowia człowieka

The claimed effect, which is proposed for further assessment, is “supports the defence against pathogens in the upper respiratory tract resulting in a decrease in duration and severity of common cold symptoms”. The proposed target population is the general adult population.
The Panel considers that the claimed effect refers to defence against pathogens in the upper respiratory tract.
The Panel considers that maintenance of upper respiratory tract defence against pathogens is a beneficial physiological effect.

3. Naukowe uzasadnienia wpływu na zdrowie człowieka - Utrzymanie odporności przeciw patogenom w górnych drogach oddechowych, prowadzące do zmniejszenia nasilenia i czasu trwania objawów przeziębienia

Five studies were provided as being pertinent for the scientific substantiation of the claim. These studies included one human intervention study (de Vrese et al., 2005), one animal study (Ohno et al., 2005) and three in vitro studies (Ghadimi et al., 2008; 2010a; 2010b).
In a randomised, double-blind, placebo-controlled parallel study, de Vrese et al. (2005) assessed the effect of consumption of the bacterial preparation on the incidence, severity and duration of common colds in 479 healthy adults (294 women aged 18-67 years). A total of 242 subjects were completed in a three-month study (from January to May 2001; 121 test group, 121 control group, 8 withdrawals) and 237 subjects participated in a 5.5 month study (from December 2001 to June 2002; 117 test group, 120 control group, 17 withdrawals). The Panel notes that these are two different studies with independent randomisation and different duration of the intervention, and that the reasons to combine data from these two cohorts are unclear. Cellular immune response was assessed in a subgroup of 122 subjects before and after 14 days of intervention in the first study. Volunteers were randomly assigned to take one tablet daily of either a bacterial preparation (containing L. gasseri PA 16/8, B. longum SP 07/3and B. bifidum MF 20/5 (5x107 CFU) with vitamins and minerals) or a control preparation (containing the same amount of vitamins and minerals only). Subjects were asked to record daily in a questionnaire the occurrence and severity of a number of symptoms, including nasal, pharyngeal, and bronchial symptoms, headache, myalgia, conjunctivitis, fatigue, loss of appetite, and fever (oral temperature >37.7°C). Common cold episodes were defined as the appearance of at least one nasal, pharyngeal or bronchial symptom, which include running nose, stuffed nose, blowing the nose, yellow secretion, bloody secretion, sneezing, scratchy throat, sore throat, hoarseness and cough. Total symptom severity scores were calculated by combining individual symptom scores over the study period. The Panel notes that no information is provided about the validity of the questionnaire used to assess the incidence, severity or duration of respiratory tract infections, that the symptoms used for the diagnosis of common cold episodes are non-specific, and that the presence of one or more of these symptoms is not an appropriate measure of respiratory tract infections in the study population. The Panel also notes that no information has been provided in the publication about the use of rescue medication, which may have confounded the results. The Panel considers that no conclusions can be drawn from this study for the scientific substantiation of the claim.
In the information provided, it is acknowledged that the mechanism by which the combination of L. gasseri PA 16/8, B. bifidum MF 20/5 and B. longum SP 07/3 could exert the claimed effect is not well understood. One animal study (Ohno et al., 2005) and three in vitro studies (Ghadimi et al., 2008; 2010a; 2010b) explored the effects of the microorganisms on the immune response to allergens/antigens. Three of these studies examined the effects of a single strain and only the in vitro
study by Ghadimi et al. (2008) assessed the effects of the combination. The Panel considers that, in the absence of evidence for an effect of the combination of L. gasseri PA 16/8, B. bifidum MF 20/5 and B. longum SP 07/3 on defence against pathogens in the upper respiratory tract in humans, these animal and in vitro studies cannot be used for the scientific substantiation of the claim as their results cannot predict the occurrence of an effect of a combination of L. gasseri PA 16/8, B. bifidum MF 20/5 and B. longum SP 07/3 in vivo in humans.
The Panel notes that no human intervention studies have been provided from which conclusions could be drawn for the scientific substantiation of the claim.
The Panel concludes that a cause and effect relationship has not been established between the consumption of a combination of Lactobacillus gasseri PA 16/8, Bifidobacterium bifidum M 20/5 and Bifidobacterium longum SP 07/3 and maintenance of upper respiratory tract defence against pathogens.

Oświadczenia zdrowotne

ID 931 - Lactobacillus gasseri PA 16/8, Bifidobacterium bifidum M 20/5, Bifidobacterium longum SP 07/3

1. Charakterystyka żywności / składnika

1.33. Characterisation of a combination of “Lactobacillus gasseri PA 16/8, Bifidobacterium bifidum MF 20/5, Bifidobacterium longum SP 07/3” (ID 931, 933)

2. Znaczenie oświadczenia dla zdrowia człowieka

3. Naukowe uzasadnienia wpływu na zdrowie człowieka - Utrzymanie odporności przeciw patogenom w górnych drogach oddechowych, prowadzące do zmniejszenia nasilenia i czasu trwania objawów przeziębienia

Warunki i możliwe ograniczenia stosowania oświadczenia