ID 960 - Bifidobacterium animalis ssp. lactis THT 010801

PL: Bifidobacterium animalis ssp. lactis THT 010801
EN: Bifidobacterium animalis ssp. lactis THT 010801
Pdf: various microorganisms

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.

1.1. Bifidobacterium animalis subsp. lactis THT 010801 (ID 960, 961, 962)

The food constituent that is the subject of the proposed health claims is Bifidobacterium animalis subsp. lactis THT 010801.
The strain B. animalis subsp. lactis THT 010801 is the trade name for B. animalis subsp. lactis LMG 18314, which is the type strain of B. animalis subsp. lactis. Culture collection numbers from different internationally recognised culture collections (e.g. LMG 18314, DSM 10140) were provided. Data on the identification and characterisation of B. animalis subsp. lactis THT 010801 at species and strain level, by using different phenotypic (carbohydrate fermentation pattern, PAGE) and genotypic (DNA- DNA hybridisation, 16S rRNA gene sequence analysis, 16S/23S intergenic spacer region sequence analysis, plasmidic profile, species-specific PCR, ARDRA, Rep-PCR, AFLP, ribotyping, MLST, RAPD and PFGE) methods, were provided (Alander et al., 2001; Crittenden et al., 2001; Duez et al., 2000; Masco et al., 2004; Matto et al., 2004; Roy and Sirois, 2000; Ventura and Zink, 2002; Ventura et al., 2006).
The Panel considers that the food constituent that is the subject of the proposed health claims, B. animalis subsp. lactis THT 010801, is sufficiently characterised.

1.44. Characterisation of “Bifidobacterium animalis spp. lactis THT 010801” (ID 960, 961, 962)

The food constituent that is the subject of the health claim is Bifidobacterium animalis spp. lactis THT 010801. No information regarding the identification/characterisation of the strain Bifidobacterium animalis ssp. lactis THT 010801 was found in the reference material provided or in the literature. It is important to point out that it may not be possible to differentiate the commercially available B. animalis ssp. lactis strains from each other on the basis of traditional genetic methods (e.g. PFGE) (Engel et al., 2003; Gueimonde et al., 2004) and may be necessary to use multi-locus sequencing or genome-wide approaches. The Panel considers that Bifidobacterium animalis ssp. lactis THT 010801, which is the subject of the health claims ID 960, 961, 962, 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.

2.1. Zmiany w funkcjach jelita (ID 960, 961, 967, 969, 971, 975, 983, 985, 994, 996, 998, 1006, 1014)

The claimed effects, which are proposed for further assessment, are: “The micro-organisms are known, for a long time, for their intestinal impact. Indeed, the bacteria promote the digestive health by
several ways. Among other thing, they degrade indigestible substances (sugars, etc.) into lactic acid and volatile fatty acids. They participate in mobility intestinal. They contribute to the secretion and absorption of nutrients”, and “The micro-organisms are known, for a long time, for their intestinal impact. Indeed, the bacteria improve the intestinal transit, especially by their participation in mobility intestinal. In more, they participate to the degradation of some indigestible substances and their absorption by the bowel.” The proposed target population is the general population.
The Panel notes that the claimed effect “contribute to the secretion and absorption of nutrients” is not sufficiently defined, and from the references provided it was not possible to establish for which nutrients an improved digestion and absorption is claimed.
The Panel assumes that the claimed effect “degrade indigestible substances (sugars, etc.) into lactic acid and volatile fatty acids” refers to changes in short chain fatty acid (SCFA) and lactic acid production. The Panel considers that changes in SCFA and lactic acid production in the gastro-intestinal tract are not beneficial physiological effects per se, but need to be linked to a beneficial physiological or clinical outcome.
The Panel assumes that the claimed effects “they participate in mobility intestinal” and “improve the intestinal transit” refer to changes in bowel function. The Panel considers that changes in bowel function such as reduced transit time, more frequent bowel movements, increased faecal bulk, or softer stools may be a beneficial physiological effect, provided that these changes do not result in diarrhoea.

2.2. Zmniejszenie ilości potencjalnie patogennych mikroorganizmów przewodu pokarmowego (ID 960, 967, 969, 971, 975, 983, 985, 994, 996, 998, 1006, 1014)

The claimed effect, which is proposed for further assessment, is: “The bacteria modulate also intestinal flora. They have a protective function by competitive inhibition on pathogen (competing for growth). They inhibit too the adhesion of these pathogens by site occupation and by production of anti- microbial substances”. The proposed target population is the general population.
The Panel assumes that the claimed effect refers to a decrease in potentially pathogenic gastro-intestinal microorganisms. The Panel considers that decreasing potentially pathogenic gastro- intestinal microorganisms might be a beneficial physiological effect.

3.1. Zmiany w funkcjach jelita (ID 960, 961, 967, 969, 971, 975, 983, 985, 994, 996, 998, 1006, 1014)

Most of the references provided in relation to these claims were on bacterial strains or combinations of strains other than those which are the subject of the claims, or on strains in combination with other substances or on microorganisms for which information on the genus only was given. Narrative reviews on the survival of bacterial strains through the stomach and small intestine, on the isolation and selection of bacterial strains, and on the potential use of recombinant dietary lactic acid bacteria for the production of oral vaccines, which were unrelated to the claim, were also provided. The Panel considers that no conclusions can be drawn from these references for the scientific substantiation of the claim.
One human intervention study on detection of the bacterial strain in faeces (Duez et al., 2000); animal or in vitro studies on induction of cytokine expression (Matsuguchi et al., 2003), inhibition of Escherichia coli urinary tract infections (Asahara et al., 2001), inhibition of Escherichia coli O157:H7 adhesion to human epithelial cells (Gagnon et al., 2004), protection against Listeria monocytogenes infection (Sato, 1984)); and/or animal and in vitro studies which addressed the potential use of a recombinant bacterial strain as carrier of proteins of immunological interest to intestinal mucosa mainly for the production of oral vaccines (Araujo Aires et al., 2006; Hazebrouck et al., 2006; Oliveira et al., 2006; Pant et al., 2006); and/or in vitro studies on the properties of the bacterial strains (e.g. survival capacity, growth at different temperatures and pH levels, viability in simulated gastro- intestinal, bile or pancreatic conditions; adhesion to human epithelial cells or to mucus from pig small intestine) (Crittenden et al., 2001; Crociani et al., 1995; Gagnon et al., 2004; Jonsson et al., 2001; Matsumoto et al., 2004; Matto et al., 2004; McMaster et al., 2005; Miyoshi et al., 2006; Todoriki et al., 2001) were also provided. The Panel notes that these studies did not address outcome measures related to the claimed effect, and considers that no conclusions can be drawn from these studies for the scientific substantiation of the claim.
The Panel notes that no human studies were provided from which conclusions could be drawn for the scientific substantiation of the claims evaluated in this section.
The Panel concludes that a cause and effect relationship has not been established between the consumption of the food constituents which are the subject of the claims evaluated in this section and changes in bowel function.

3.2. Zmniejszenie ilości potencjalnie patogennych mikroorganizmów przewodu pokarmowego (ID 960, 967, 969, 971, 975, 983, 985, 994, 996, 998, 1006, 1014)

Most of the references provided in relation to these claims were on bacterial strains or combinations of strains other than those which are the subject of the claims, or on strains in combination with other substances. Narrative reviews on the survival of bacterial strains through the stomach and small intestine, on the isolation and selection of bacterial strains, and on the potential use of recombinant dietary lactic acid bacteria for the production of oral vaccines, which were unrelated to the claim, were also provided. The Panel considers that no conclusions can be drawn from these references for the scientific substantiation of the claim.
Animal studies on induction of cytokine expression (Matsuguchi et al., 2003), on inhibition of Escherichia coli urinary tract infections (Asahara et al., 2001), and/or in vitro studies on the properties of the bacterial strains (i.e. survival capacity, growth at different temperatures and pH levels, viability in simulated gastro-intestinal, bile or pancreatic conditions; adhesion to human epithelial cells or to mucus from pig small intestine) (Crittenden et al., 2001; Crociani et al., 1995; Gagnon et al., 2004; Jonsson et al., 2001; Matsumoto et al., 2004; Matto et al., 2004; McMaster et al., 2005; Miyoshi et al., 2006; Todoriki et al., 2001), were also provided. In vitro and in vivo studies in animals which addressed the potential use of recombinant bacterial strains as carriers of proteins of immunological interest to intestinal mucosa mainly for the production of oral vaccines (Araujo Aires et al., 2006; Hazebrouck et al., 2006; Oliveira et al., 2006; Pant et al., 2006) were provided for ID 975. The Panel notes that these studies did not address outcome measures related to the claimed effect, and considers that no conclusions can be drawn from these studies for the scientific substantiation of the claim.
The Panel notes that no human studies were provided from which conclusions could be drawn for the scientific substantiation of the claims evaluated in this section.
For ID 971, an in vitro study investigated the inhibition of Escherichia coli O157:H7 adhesion to a human epithelial cell line in the presence of the bacterial strain that is the subject of the claim (Gagnon et al., 2004). For ID 975, a study in animals investigated the protective activity of the bacterial strain that is the subject of the claim against Listeria monocytogenes infection (Sato, 1984). The Panel considers that in the absence of evidence for an effect on decreasing potentially pathogenic gastro- intestinal microorganisms in humans, evidence provided in animal and in vitro studies cannot be used alone for the scientific substantiation of a claim on decreasing potentially pathogenic gastro-intestinal microorganisms.
The Panel concludes that a cause and effect relationship has not been established between the consumption of the food constituents which are the subject of the claims evaluated in this section and decreasing potentially pathogenic gastro-intestinal microorganisms.

Warunki i możliwe ograniczenia stosowania oświadczenia

At least 108 CFU/day