SCIENTIFIC OPINION Scientific Opinion on the safety and efficacy of Actisaf Sc47 (Saccharomyces cerevisiae) as a feed additive for rabbits for fattening and non foodproducing rabbits 1, 2 EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) 3,4 ABSTRACT European Food Safety Authority (EFSA), Parma, Italy The additive Actisaf Sc47 is a microbiological feed additive consisting only of dried cells of Saccharomyces cerevisiae. The applicant has requested the re-evaluation of the product when used as a zootechnical additive (functional group: gut flora stabilisers) in diets for rabbits for fattening and the extension of use in diets for non food-producing rabbits at the inclusion rate of 5 x 10 9 CFU/kg feedingstuffs. S. cerevisiae is a species which is considered by EFSA to be suitable for the QPS approach to safety assessment. As the identity of the active agent has been established, the use of Actisaf Sc47 can be presumed safe for the target species, consumers and the environment. Actisaf Sc47 is non-irritant to skin. Given the absence of particles of inhalable size and the virtual absence of dust, exposure and therefore sensitisation via a respiratory route is not to be expected. Three trials performed with weaned rabbits showed the potential of Actisaf Sc47 to reduce mortality when used in diets at a dose of 5 x 10 9 CFU/kg feed. A similar effect can be expected in non food-producing rabbits. European Food Safety Authority, 2012 KEY WORDS Zootechnical additive, gut flora stabiliser, Actisaf Sc47, Saccharomyces cerevisiae, rabbits for fattening, non food-producing rabbits, safety, efficacy 1 On request from the European Commission, Question No EFSA-Q-2010-00936, adopted on 13 December 2011. 2 This scientific opinion has been edited following the provisions of Article 8(6) and Article 18 of Regulation (EC) No 1831/2003. The modified sections are indicated in the text. 3 Panel members: Gabriele Aquilina, Georges Bories, Andrew Chesson, Pier Sandro Cocconcelli, Joop de Knecht, Noël Albert Dierick, Mikolaj Antoni Gralak, Jürgen Gropp, Ingrid Halle, Christer Hogstrand, Reinhard Kroker, Lubomir Leng, Secundino López Puente, Anne-Katrine Lundebye Haldorsen, Alberto Mantovani, Giovanna Martelli, Miklós Mézes, Derek Renshaw, Maria Saarela, Kristen Sejrsen and Johannes Westendorf. Correspondence: FEEDAP@efsa.europa.eu 4 Acknowledgement: The Panel wishes to thank the members of the Working Group on Micro-organisms, including Joaquim Brufau and Guido Rychen, for the preparatory work on this scientific opinion. Suggested citation: EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP); Scientific Opinion on the safety and efficacy of Actisaf Sc47 (Saccharomyces cerevisiae) as a feed additive for rabbits for fattening and non food-producing rabbits.. [12 pp.]. doi:10.2903/j.efsa.2012.2531. Available online: www.efsa.europa.eu/efsajournal European Food Safety Authority, 2012
SUMMARY Following a request from the European Commission, the Panel on Additives an Products or Substances used in Animal Feed (FEEDAP) was asked to deliver a scientific opinion on the safety and efficacy of the product Actisaf Sc47 as a feed additive for rabbits for fattening and non food-producing rabbits. The applicant has requested the re-evaluation of the product when used as a zootechnical additive (functional group: gut flora stabilisers) in diets for rabbits for fattening and the extension of use in diets for non food-producing rabbits at the inclusion rate of 5 x 10 9 CFU/kg feedingstuffs. Actisaf Sc47 consists only of dried cells of Saccharomyces cerevisiae, a species which is considered by EFSA to be suitable for the qualified presumption of safety (QPS) approach to safety assessment. As the identity of the active agent has been established, the use of Actisaf Sc47 can be presumed safe for the target species, consumers of products derived from these food-producing species and the environment. Actisaf Sc47 is non-irritant to skin. Given the absence of particles of inhalable size and the virtual absence of dust, exposure and therefore toxicity and sensitisation via a respiratory route is not to be expected. Three trials performed with weaned rabbits showed the potential of Actisaf Sc47 to reduce mortality when used in diets at a minimum dose of 5 x 10 9 CFU/kg feed. A similar effect can be expected in non food-producing rabbits. 2
TABLE OF CONTENTS Abstract... 1 Summary... 2 Table of contents... 3 Background... 4 Terms of reference... 5 Assessment... 7 1. Introduction... 7 2. Characterisation... 7 2.1. Characterisation of the active agent... 7 2.2. Characterisation of the product... 7 2.3. Stability and homogeneity... 8 2.4. Conditions of use... 8 2.5. Evaluation of the analytical methods by the European Union Reference Laboratory (EURL) 8 3. Safety... 9 3.1. Safety for the user... 9 4. Efficacy... 9 4.1. Efficacy for rabbits for fattening... 9 4.2. Efficacy for non food-producing rabbits... 10 5. Post-market monitoring... 10 Documentation provided to EFSA... 10 References... 10 Appendix... 12 3
BACKGROUND Regulation (EC) No 1831/2003 5 establishes the rules governing the European Union authorisation of additives for use in animal nutrition. In particular, Article 4(1) of that Regulation lays down that any person seeking authorisation for a feed additive or for a new use of a feed additive shall submit an application in accordance with Article 7; in addition, Article 10(2) of that Regulation also specifies that for existing products within the meaning of Article 10(1), an application shall be submitted in accordance with Article 7, at the latest one year before the expiry date of the authorisation given pursuant to Directive 70/524/EEC for additives with a limited authorisation period, and within a maximum of seven years after the entry into force of this Regulation for additives authorised without time limit or pursuant to Directive 82/471/EEC. The European Commission received a request from the company Lesaffre Feed Additives 6 for authorisation and re-evaluation of the product Actisaf Sc47 (Saccharomyces cerevisiae CNCM I- 4407), when used as a feed additive for rabbits for fattening and non food-producing rabbits (category: zootechnical additive; functional group: gut flora stabiliser) under the conditions mentioned in Table 1. According to Article 7(1) of Regulation (EC) No 1831/2003, the Commission forwarded the application to the European Food Safety Authority (EFSA) as an application under Article 4(1) (authorisation of a feed additive or new use of a feed additive) and under Article 10(2)/(7) (reevaluation of an authorised feed additive). EFSA received directly from the applicant the technical dossier in support of this application. 7 According to Article 8 of that Regulation, EFSA, after verifying the particulars and documents submitted by the applicant, shall undertake an assessment in order to determine whether the feed additive complies with the conditions laid down in Article 5. The particulars and documents in support of the application were considered valid by EFSA as of 29 September 2010. The additive Actisaf Sc47 is a preparation of Saccharomyces cerevisiae CNCM I-4407. This product is already authorised for use in cattle for fattening, 8 sows, 9 piglets, 10 rabbits for fattening, 11 dairy cows, 12 lambs for fattening, 13 dairy goats and dairy sheep, 14 horses, 15 pigs for fattening, 16 dairy buffaloes 17 and calves for rearing. 18 5 Regulation (EC) No 1831/2003 of the European Parliament and of the Council of 22 September 2003 on additives for use in animal nutrition. OJ L 268, 18.10.2003, p. 29. 6 Lesaffre Feed Additives, a division of S.I. Lesaffre, 137 rue Gabriel Péri, 59700, Marcq-en-Baroeul, France. 7 EFSA Dossier reference: FAD-2010-0038. 8 Commission Regulation (EC) No 316/2003 of 19 February 2003 concerning the permanent authorisation of an additive in feedingstuffs and the provisional authorisation of a new use of an additive already authorised in feedingstuffs. OJ L 46, 20.2.2003, p. 15. 9 Commission Regulation (EC) No 1288/2004 of 14 July 2004 concerning the permanent authorisation of certain additives and the provisional authorisation of a new use of an additive already authorised in feedingstuffs. OJ L 243, 15.7.2004, p. 10. 10 Commission Regulation (EC) No 2148/2004 of 16 December 2004 concerning the permanent and provisional authorisations of certain additives and the authorisation of new uses of an additive already authorised in feedingstuffs. OJ L 370, 17.12.2004, p. 24. 11 Commission Regulation (EC) No 600/2005 of 18 April 2005 concerning a new authorisation for 10 years of a coccidiostat as an additive in feedingstuffs, the provisional authorisation of an additive and the permanent authorisation of certain additives in feedingstuffs. OJ L 99, 19.4.2005, p. 5. 12 Commission Regulation (EC) No 1811/2005 of 4 November 2005 concerning the provisional and permanent authorisations of certain additives in feedingstuffs and the provisional authorisation of a new use of an additive already authorised in feedingstuffs. OJ L 291, 5.11.2005, p. 12. 13 Commission Regulation (EC) No 1447/2006 of 29 September 2006 concerning the authorisation of a new use of Saccharomyces cerevisiae (Biosaf SC 47) as a feed additive. OJ L 271, 30.9.2006, p. 28. 14 Commission Regulation (EC) No 188/2007 of 23 February 2007 concerning the authorisation of a new use of Saccharomyces cerevisiae (Biosaf SC 47) as a feed additive. OJ L 57, 24.2.2007, p. 3. 15 Commission Regulation (EC) No 186/2007 of 21 February 2007 concerning the authorisation of a new use of Saccharomyces cerevisiae (Biosaf SC 47) as a feed additive. OJ L 63, 1.3.2007, p. 6. 4
The Scientific Committee on Animal Nutrition (SCAN) issued an opinion on the safety of this product for the piglets, sows, beef and dairy cattle, including the safety for the user, the consumer and the environment (EC, 1997/2003), and another on the efficacy for cattle for fattening (EC, 2002). EFSA issued several opinions on the safety and efficacy of this product for the following species: lambs for fattening (EFSA, 2006a), dairy small ruminants (EFSA, 2006b), horses (EFSA, 2006c), calves for rearing (EFSA, 2007a), pigs for fattening (EFSA, 2007b), and dairy buffaloes (EFSA, 2008), and one on the efficacy for calves for rearing (EFSA, 2010a). TERMS OF REFERENCE According to Article 8 of Regulation (EC) No 1831/2003, EFSA shall determine whether the feed additive complies with the conditions laid down in Article 5. EFSA shall deliver an opinion on the safety for the target animals, consumer, user and the environment and the efficacy of the product Actisaf Sc47 (Saccharomyces cerevisiae NCYC Sc47) when used under the conditions described in Table 1. 16 Commission Regulation (EC) No 209/2008 of 6 March 2008 concerning the authorisation of a new use of Saccharomyces cerevisiae (Biosaf Sc 47) as a feed additive. OJ L 63, 7.3.2008, p. 3. 17 Commission Regulation (EC) No 232/2009 of 19 March 2009 concerning the authorisation of a new use of Saccharomyces cerevisiae NCYC Sc47 as a feed additive for dairy buffaloes (holder of the authorisation Société Industrielle Lesaffre). OJ L 74, 20.3.2009, p. 14. 18 Commission Regulation (EU) No 883/2010 of 7 October 2010 concerning the authorisation of a new use of Saccharomyces cerevisiae NCYC Sc 47 as a feed additive for calves for rearing (holder of the authorisation Société industrielle Lesaffre). OJ L 265, 8.10.2010, p. 1. 5
Table 1: Description and conditions of use of the additive as proposed by the applicant Additive Registration number/ec No/No Category of additive Functional group of additive Saccharomyces cerevisiae NCYC Sc47 4b1702 Zootechnical additives Gut flora stabiliser Description Chemical Purity criteria Method of analysis Composition, description formula Saccharomyces cerevisisae - - - Trade name Name of the holder of authorisation Actisaf Sc47 Lesaffre Feed Additive, a division of S.I.Lesaffre Conditions of use Species or category of animal Rabbits for fattening, pet rabbits and non food-producing rabbits Maximum Age Minimum content Maximum content Withdrawal period CFU/kg of complete feedingstuffs - 5 x 10 9 - - Other provisions and additional requirements for the labelling Specific conditions or restrictions for use Specific conditions or restrictions for handling Post-market monitoring In the directions for use of the additive and premixture, indicate the storage temperature, storage life and stability to pelleting - - Specific conditions for use in complementary feedingstuffs - Marker residue Maximum Residue Limit (MRL) Species or category of animal Target tissue(s) or food products Maximum content in tissues - -- - - 6
ASSESSMENT 1. Introduction Actisaf Sc47 19 is the trade name for a feed additive based on viable cells of a strain of Saccharomyces cerevisiae. The product is currently authorised for use with cattle for fattening, sows, piglets, rabbits for fattening, dairy cows, lambs for fattening, dairy goats and dairy sheep, horses, pigs for fattening, dairy buffaloes and calves for rearing (see Background). The applicant has applied for the reevaluation of the product when used as a zootechnical additive (functional group: gut flora stabilisers) in diets for rabbits for fattening and for the authorisation for use in diets for non food-producing rabbits. Saccharomyces cerevisiae is considered by the European Food Safety Authority (EFSA) to be suitable for the Qualified Presumption of Safety (QPS) approach to safety assessment (EFSA, 2010b). Therefore, no assessment of safety for the target species, consumer and the environment is required provided that the identity of the strain can be established. 2. Characterisation 2.1. Characterisation of the active agent The active agent of Actisaf is Saccharomyces cerevisiae strain CNCM I-4407. The strain is deposited at the Collection Nationale de Cultures de Microoganismes. 20 It has been identified based on morphological and phenotypic properties. It has not been subject to any genetic modification. The genetic stability of the strain (three samples) has been shown with pulsed field gel electrophoresis during 1990 2000. 21 2.2. Characterisation of the product 22 Actisaf Sc47 consists only of dried yeast cells. It is produced by batch fermentation in a growth medium typical of those used for the industrial production of yeasts. The final product is produced in three different forms: standard (STD), powder and heat resistant (Hr+). The powder form is a fraction of the STD obtained by sieving the dry substance between 217 and 500 µm. The Hr+ form is produced with a modified drying protocol which leads to a product with a lower humidity content and a lower water activity than the standard. The minimum of number of viable cells is 5 x 10 9 CFU/g. Batch to batch variation has been measured on 19 of the STD form batches during 2004 2009. 23 The viable yeast numbers varied between 8 x 10 9 and 1.2 x 10 10 CFU/g. Particle size distribution has been determined by sieving a single batch of powder and STD forms, and four batches of the Hr+ form. 24 For Actisaf Sc47 STD, 78 % of the particles were larger than 800 µm and none were smaller than 125 µm. For Actisaf Sc47 powder all particles were smaller than 500 µm; 9 % were in the range of 80 to 125 µm and 0.3 % were smaller than 80 µm. The particle size distribution of the Hr+ form mirrored that of the STD form. The dusting potential of the three product forms was determined by Heubach dustmeter (three measurements per form) and shown to be essentially zero in all three cases. 25 The additive is tested annually for heavy metals and contaminants. Chemical purity data concerning heavy metals (Cd, Pb, Hg), As, F and aflatoxin B1 were provided for seven batches of the STD form, 19 The products was previously traded with the name Biosaf Sc47. 20 Technical dossier/ Supplementary info Nov 11. 21 Technical dossier/section II. 22 This section has been edited following the provisions of Article 8(6) and Article 18 of Regulation (EC) No 1831/2003. 23 Technical dossier/section II. 24 Technical dossier/section II/Appendix II-1-c. 25 Technical dossier/supplementary info July 11/Appendix II-2. 7
two batches of the Hr+ form and a single batch of the powder form. All heavy metal and aflatoxin levels were below the limit levels set by the Company. These reflect existing legal limits. 26 Three batches of the STD and Hr+ forms were studied for microbial contamination. 24 The levels of total coliforms and Escherichia coli were < 10 CFU/g, Staphylococcus aureus was not detected in 1 g of the additive and Salmonella spp. was absent in 25 g of the additive. 2.3. Stability and homogeneity Shelf life has been tested on three batches of the STD form (at 26 o C), three batches of the powder form (25 o C/60% RH and 30 o C/65 % RH) and four batches of the Hr+ form (two batches at 25 o C/60 % RH, and two other batches under room conditions ) stored in commercial packaging up to 12 18 months. 27 After 12 and 18 months of storage, the STD and powder forms, respectively, showed essentially no loss of viability. The two batches of the Hr+ form tested at room conditions were stable for 14 months, while the two Hr+ batches tested at 25 o C/60 % RH were stable for 12 months. 28 Stability to pelleting was tested with a variety of feeds (including feed for rabbits) under a range of conditioning and die temperatures up to 90 o C. Four batches of the STD form and five batches of the Hr+ form were used. 29 Both forms survived pelleting at temperatures up to 80 o C with the Hr+ form showing slightly better stability (up to 85 o C). The stability of the three Actisaf Sc47 forms (two batches of each) was studied in mineral/vitamin premixtures for sows. 30 The STD and powder forms showed no loss of viability after a nine month storage at 20 o C, while the Hr+ form showed a small loss (0.3-0.4 Log-values) when stored under the same conditions. Data specific for a mineral/vitamin premixture for rabbits was only provided for three months for an unspecified form of the additive. No losses were observed over this period. 31 When mixed with rabbit feed and stored in atmospheric conditions at appr. 20 o C for three months, two batches of the STD form showed viability losses up to 0.5 Log-values. 32 The stability of three different batches of each of the three forms was also studied in piglet feed. 33 All three forms were stable over the three-month storage period. The ability of Actisaf Sc47 STD to form homogeneous mixtures was studied in a rabbit feed (based on oats and other cereals, alfalfa, sunflower meal and molasses). The coefficient of variation of eight subsamples was 2.4 %. 34 2.4. Conditions of use The product is intended for use in feeds for rabbits for fattening and non food-producing rabbits at the dose of 5 x 10 9 CFU/kg feedingstuffs. 2.5. Evaluation of the analytical methods by the European Union Reference Laboratory (EURL) EFSA has verified the EURL report as it relates to the methods used for the control of the active agent in animal feed. The Executive Summary of the EURL report can be found in the Appendix. 26 Technical dossier/supplementary info July 11/Appendix II-1. 27 Technical dossier/section II/Appendix II-4-a, b, c. 28 Technical dossier/supplementary info July 11/Appendix II-5. 29 Technical dossier/section II/Appendix II-4-c and Supplementary info July 11/Appendix II-6. 30 Technical dossier/supplementary info July 11/Appendix II-7. 31 Technical dossier/section II/Appendix II-4-d. 32 Technical dossier/section II/Appendix II-4-f. 33 Technical dossier/supplementary info July 11/Appendix II-9. 34 Technical dossier/section II/Appendix II-4-h. 8
3. Safety Actisaf Sc47 for rabbits for fattening and pet rabbits Saccharomyces is considered by EFSA to be suitable for the QPS approach to safety assessment (EFSA, 2010b). In the view of the FEEDAP Panel, the identity of the production strain is established as Saccharomyces and therefore, no further assessment of safety for the target species, consumer and the environment is required. 3.1. Safety for the user An acute skin irritation study in the rabbit was performed according to OECD Guideline 404. Under the experimental conditions used, Actisaf Sc47 STD was found to be non-irritant for the skin of the rabbit. 35 Eye irritation and skin sensitisation were not tested. Given the absence of particles of inhalable size and the virtual absence of dust, exposure and therefore toxicity and sensitisation via a respiratory route are not to be expected. 4. Efficacy The applicant provided six studies performed in three different Member States. However, three studies were not considered, one due to the very high mortality observed (nearly 50 % in the control group) 36 and two because of insufficient reporting (e.g. absence of raw data, no indication of the number of replicates, limited statistics). 37 4.1. Efficacy for rabbits for fattening Trials 1 and 2 shared the same experimental design (44 days) but were conducted in two different breeding farms. Each trial consisted of two experimental groups (control and Actisaf Sc47 at 5 x 10 9 CFU/kg feed). 38 Trial 1 was performed on six consecutive weanings, giving a total number of 891 rabbits (425 rabbits were assigned to the yeast group). Distribution between the two treatments was randomly made at weaning. Trial 2 included eight consecutive weanings giving a total number of 1216 animals randomly distributed to the two experimental groups (608 kits per treatment). In both trials the Actisaf Sc47 dose in feed was confirmed by analysis. The performance parameters have been analysed by variance analysis and mortality of rabbits by Chi 2 (SPSS software). Results indicated: - Trial 1: a significant effect on mortality (19 % in the control group, 10 % in the Actisaf Sc47 group, 89/466 vs 43/425, P < 0.05) but no significant effects on growing parameters. - Trial 2: a significant effect on mortality (10 % in the control group, 6.5 % in the Actisaf Sc47 group, 61/608 vs 40/608, P < 0.05) but no significant effects on growing parameters. The third trial (42 days) was performed with 1000 weaned rabbits placed in cages (five animals per cage). 39 Two homogeneous groups were constituted (100 cages) containing 500 animals per group (control and Actisaf Sc47 STD at 5 x 10 9 CFU/kg, confirmed by analysis). Feed intake and the weight of the animals were recorded at the cage level. The data were analysed using one-way ANOVA and the cage as experimental unit). The results indicated a significant improvement in body weight (2.38 kg vs 2.03 kg, P < 0.05) and feed conversion ratio (3.11 vs 3.28, P < 0.05) and a reduced mortality (11 % (55/499) vs 25 % (125/499), P < 0.05) in the Actisaf Sc47 group when compared to untreated animals. 35 Technical dossier/supplementary info July 11/Appendix III-1. 36 Technical dossier/section IV/Annex IV_4. 37 Technical dossier/section IV/Annex IV_1. 38 Technical dossier/section IV/Annex IV_2 and Supplementary info July 11/Appendix IV-1. 39 Technical dossier/section IV/Annex IV_3. 9
4.2. Efficacy for non food-producing rabbits Actisaf Sc47 for rabbits for fattening and pet rabbits The dose proposed for use with the non food-producing rabbits is the same as that demonstrated effective in rabbits for fattening and it can be reasonably assumed that the mode of action is the same. The only consistent effect in rabbits for fattening is reduced mortality, which is also an appropriate endpoint for non food-producing rabbits. 5. Post-market monitoring The FEEDAP Panel considers that there is no need for specific requirements for a post-market monitoring plan other than those established in the Feed Hygiene Regulation 40 and Good Manufacturing Practice. CONCLUSIONS On the basis of the QPS approach, the product can be presumed safe for rabbits for fattening and non food-producing rabbits, for consumers of products derived from these food-producing species and for the environment. Actisaf Sc47 is non irritant to skin. Given the absence of particles of inhalable size and the virtual absence of dust, exposure and therefore, toxicity and sensitisation via a respiratory route is not to be expected. Actisaf Sc47 showed the potential to reduce mortality in rabbits for fattening at a minimum dose of 5 x 10 9 CFU/kg feed. A similar effect can be expected in non food-producing rabbits. Conclusions on the safety and efficacy of Actisaf Sc47 apply to the three forms. DOCUMENTATION PROVIDED TO EFSA 1. Actisaf Sc47 for rabbits for fattening. June 2010. Submitted by Lesaffre Feed additives. 2. Actisaf Sc47 for rabbits for fattening. Supplementary information. July 2011. Submitted by Lesaffre Feed additives. 3. Actisaf Sc47 for rabbits for fattening. Supplementary information. November 2011. Submitted by Lesaffre Feed additives. 4. Evaluation report of the European Union Reference Laboratory for Feed Additives on the methods(s) of analysis for Actisaf Sc47 for rabbits 5. Comments from Member States received through the ScienceNet. REFERENCES EC (European Commission), 1997, updated 2003, online. Scientific Committee on Animal Nutrition. Opinion on the use of certain micro-organisms as additives in feedingstuffs. Available from http://ec.europa.eu/food/fs/sc/scan/out93_en.pdf EC (European Commission), 2002, online. Scientific Committee on Animal Nutrition. Evaluation of the efficacy of micro-organism product Biosaf Sc 47. Available from http://ec.europa.eu/food/fs/sc/scan/out101_en.pdf EFSA (European Food Safety Authority), 2006a. Opinion of the Scientific Panel on additives and products or substances used in animal feed (FEEDAP) on the safety and efficacy of the product Biosaf Sc47, a preparation of Saccharomyces cerevisiae (NCYC Sc47), as a feed additive for lambs in accordance with Regulation (EC) No 1831/2003. The EFSA Journal, 321, 1-8. 40 Regulation (EC) No 183/2005 of the European Parliament and of the Council of 12 January 2005 laying down requirements for feed hygiene. OJ L 35, 8.2.2005, p. 1. 10
EFSA (European Food Safety Authority), 2006b. Scientific Opinion of the Panel on additives and products or substances used in animal feed (FEEDAP) on the safety and efficacy of the product Biosaf Sc47, a preparation of Saccharomyces cerevisiae, as a feed additive for dairy small ruminants. The EFSA Journal, 379, 1-9. EFSA (European Food Safety Authority), 2006c. Scientific Opinion of the Panel on additives and products or substances used in animal feed on the safety and efficacy of the product Biosaf Sc47, a preparation of Saccharomyces cerevisiae, as a feed additive for horses. The EFSA Journal, 384, 1-9. EFSA (European Food Safety Authority), 2007a. Scientific Opinion of the Scientific Panel on additives and products or substances used in animal feed (FEEDAP) on the safety and efficacy of Biosaf Sc47, a preparation of Saccharomyces cerevisiae, as a feed additive for calves for rearing according to Regulation (EC) No 1831/2003. The EFSA Journal, 459, 1-8. EFSA (European Food Safety Authority), 2007b. Scientific Opinion of the Panel on Additives and Products or Substances used in Animal Feed on the safety and efficacy of Biosaf Sc47 (Saccharomyces cerevisiae) as feed additive for pigs for fattening. The EFSA Journal, 585, 1-9. EFSA (European Food Safety Authority), 2008. Scientific Opinion of the Panel on Additives and Products or Substances used in Animal Feed on the safety and efficacy of Biosaf Sc47 (Saccharomyces cerevisiae NCYC Sc47) as a feed additive for dairy buffaloes. The EFSA Journal, 837, 1-10. EFSA (European Food Safety Authority), 2010a. EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP). Scientific Opinion on the efficacy of Biosaf Sc47 (Saccharomyces cerevisiae) for calves for rearing. EFSA Journal, 8(4): 1576. EFSA (European Food Safety Authority), 2010b. Scientific Opinion on the maintenance of the list of QPS microorganisms intentionally added to food or feed (2010 update). EFSA Panel on Biological Hazards (BIOHAZ). EFSA Journal, 8(12):1944. 11
APPENDIX Executive Summary of the Evaluation Report of the European Union Reference Laboratory for Feed Additives on the Method(s) of Analysis for Actisaf Sc47 41 In the current application authorisation is sought under articles 4(1) and 10(2) for feed additive Actisaf Sc47 under the category 'zootechnical additives', functional group 4(b) 'gut flora stabilisers', according to Annex I of Regulation (EC) No 1831/2003. Specifically, the authorisation is sought for the use of Actisaf Sc47 for rabbits for fattening, pet rabbits and non food-producing rabbits. The feed additive is intended to be marketed in three different forms as Actisaf Sc47 Powder, Actisaf Sc47 Standard and Actisaf Sc47 Hr+. The active component of Actisaf Sc47 is a pure culture of the strain Saccharomyces cerevisiae NCYC Sc47, with a minimum concentration of 5 x 10 9 CFU/g. It is intended to be mixed to complete feedingstuffs at a minimum dose of 5 x 10 9 CFU/kg. For the enumeration of Saccharomyces cerevisiae NCYC Sc47 in feed additive, premixtures and feedingstuffs the Applicant proposes the ring trial validated CEN pour plate method for the enumeration of yeast probiotic strains (EN 15789), using yeast extract dextrose chloramphenicol agar (CGYE). The performance characteristics of the EN 15789 method reported after logarithmic transformation (CFU) are: - a repeatability standard deviation (s r ) ranging from 0.17 to 0.36 log 10 CFU/g, - a reproducibility standard deviation (s R ) ranging from 0.55 to 0.60 log 10 CFU/g; and - a limit of detection (LOD) of 1 x 10 5 CFU/kg, well below the minimum dose proposed by the applicant (5 x 10 9 CFU/kg of feedingstuffs). Based on these performance characteristics the CRL recommends, for official control, the CEN method EN 15789 for the enumeration of Saccharomyces cerevisiae NCYC Sc47 in feed additives, premixtures and feedingstuffs. Molecular methods were used by the Applicant for identification of the active agent. The EURL recommends for official control Polymerase Chain Reaction (PCR), a generally recognised standard methodology for identification of yeasts. Further testing or validation of the methods to be performed through the consortium of National Reference Laboratories as specified by article 10 (Commission Regulation (EC) No 378/2005) is not considered necessary. 41 The full report is available from the EURL website : http://irmm.jrc.ec.europa.eu/sitecollectiondocuments/finrep-fad- 2010-0038.pdf 12