EFSA Journal 2015;13(5):4106 SCIENTIFIC OPINION Scientific Opinion on the safety and efficacy of Rovabio Spiky (endo-1,4- beta-xylanase and endo-1,3(4)-beta-glucanase) as a feed additive for turkeys and minor poultry species for fattening or reared for laying or breeding 1 EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) 2,3 ABSTRACT European Food Safety Authority (EFSA), Parma, Italy Rovabio Spiky is an enzyme preparation, available in solid and liquid forms, of endo-1,4-beta-xylanase and endo-1,3(4)-beta-glucanase. The enzymes present in the additive are produced by two strains of Penicillium funiculosum, one of which is genetically modified. The additive is intended to be used as a feed additive for turkeys for fattening and minor poultry species for fattening or reared for laying or breeding. Aspects other than the safety and efficacy for turkeys have been addressed in the previous opinion on the safety and efficacy for chickens for fattening, chickens reared for laying and other minor poultry species. The current assessment addresses these aspects for the new target species only. Based on the results of a tolerance trial in turkeys for fattening, the Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) concludes that the additive is safe for turkeys for fattening under the recommended conditions of use. Based on the results obtained in three efficacy studies, the FEEDAP Panel concludes that the additive has the potential to be efficacious in turkeys for fattening at the minimum recommended dose (1 100 U/kg xylanase and 760 U/kg glucanase). Both conclusions are extended to turkeys reared for breeding. The FEEDAP Panel confirms the previous conclusions that the additive is safe and efficacious in minor poultry species for fattening, reared for laying and reared for breeding at the same conditions of use. European Food Safety Authority, 2015 KEY WORDS zootechnical additive, digestibility enhancers, endo-1,4-beta-xylanase, endo-1,3(4)-beta-glucanase, turkeys, minor poultry species 1 On request from the European Commission, Question No EFSA-Q-2014-00463, adopted on 28 April 2015. 2 Panel members: Gabriele Aquilina, Vasileios Bampidis, Maria De Lourdes Bastos, Lucio Guido Costa, Gerhard Flachowsky, Mikolaj Antoni Gralak, Christer Hogstrand, Lubomir Leng, Secundino López-Puente, Giovanna Martelli, Baltasar Mayo, Fernando Ramos, Derek Renshaw, Guido Rychen, Maria Saarela, Kristen Sejrsen, Patrick Van Beelen, Robert John Wallace and Johannes Westendorf. Correspondence: FEEDAP@efsa.europa.eu 3 Acknowledgement: The Panel wishes to thank the members of the Working Group on Enzymes, including Paul Brantom, Noël Dierick and Ingrid Halle, for the preparatory work on this scientific opinion. Suggested citation: EFSA FEEDAP Panel (EFSA Panel on Additives and Products or Substances used in Animal Feed), 2015. Scientific Opinion on the safety and efficacy of Rovabio Spiky (endo-1,4-beta-xylanase and endo-1,3(4)-betaglucanase) as a feed additive for turkeys and minor poultry species for fattening, reared for laying or reared for breeding. EFSA Journal 2015;13(5):4106, 9 pp. doi:10.2903/j.efsa.2015.4106 Available online: www.efsa.europa.eu/efsajournal European Food Safety Authority, 2015
SUMMARY Following a request from the European Commission, the Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) was asked to deliver a scientific opinion on the safety and efficacy of Rovabio Spiky (endo-1,4-beta-xylanase and endo-1,3(4)-beta-glucanase) as a feed additive for turkeys and minor poultry species for fattening or reared for laying or breeding. This additive is prepared by combining two products of fermentation obtained from two strains of Penicillium funiculosum (Talaromyces versatilis sp. nov.), one of which is non-genetically modified and the other is genetically modified. The additive is available in solid and liquid forms. Both formulations are considered equivalent in terms of safety and efficacy for the target species. Aspects other than the safety and efficacy for turkeys have been addressed in the previous opinion on the safety and efficacy of Rovabio Spiky as a feed additive for chickens for fattening, chickens reared for laying and other minor poultry species. Therefore, the present assessment will address these aspects for the new target species only. Based on the results obtained in the tolerance trial in turkeys for fattening, the FEEDAP Panel concludes that the additive is safe for turkeys for fattening under the recommended conditions of use. This conclusion is extended to turkeys reared for breeding. Based on the results obtained in three long-term trials, the FEEDAP Panel concludes that the additive has the potential to be efficacious in turkeys for fattening at the minimum recommended dose (1 100 U/kg xylanase and 760 U/kg glucanase). This conclusion is extended to turkeys reared for breeding. The FEEDAP Panel confirms the previous conclusions that the additive is safe and efficacious in minor poultry species for fattening, reared for laying and reared for breeding at the same conditions of use. EFSA Journal 2015; 13(5):4106 2
TABLE OF CONTENTS Abstract... 1 Summary... 2 Background... 4 Terms of reference... 4 Assessment... 7 1. Introduction... 7 2. Conditions of use... 7 3. Evaluation of the analytical methods by the European Union Reference Laboratory (EURL)... 7 4. Safety for the target species... 7 5. Efficacy... 8 6. Post-market monitoring... 9 Conclusions... 9 Documentation provided to EFSA... 9 References... 9 EFSA Journal 2015; 13(5):4106 3
BACKGROUND Regulation (EC) No 1831/2003 4 establishes the rules governing the Community 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. The European Commission received a request from ADISSEO France SAS 5 for authorisation of the product Rovabio Spiky, endo-1,4-beta-xylanase and endo-1,3(4)-beta-glucanase, when used as a feed additive for major and minor poultry species for fattening, reared for laying and breeding (category: zootechnical additive; functional group: digestibility enhancers) 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). EFSA received directly from the applicant the technical dossier in support of this application. 6 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 1 August 2014. The additive Rovabio Spiky is a preparation of endo-1,4-beta-xylanase and endo-1,3(4)-betaglucanase produced by combining two products of fermentation obtained from two strains of Talaromyces versatilis (IMI 378536 and DSM 26702). The Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) issued an opinion on the safety and efficacy of Rovabio Spiky (endo-1,4-beta-xylanase and endo-1,3(4)-beta-glucanase) as a feed additive for chickens for fattening, chickens reared for laying and other minor poultry species (for fattening and reared for laying) (EFSA FEEDAP Panel, 2014). 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 Rovabio Spiky (endo-1,4-beta-xylanase and endo-1,3(4)-beta-glucanase), when used under the conditions described in Table 1. 4 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. 5 ADISSEO France SAS, Immeuble Antony Parc II 10 place du Général de Gaulle 92160 Antony, France. 6 EFSA Dossier reference: FAD-2014-0020. EFSA Journal 2015; 13(5):4106 4
Table 1: Description and conditions of use of the additive as proposed by the applicant Additive Registration number/ec No/No Category(-ies) of additive Zootechnical additive (4) Endo-1,4-beta-xylanase EC 3.2.1.8 and Endo-1,3(4)-beta-glucanase EC 3.2.1.6 - Functional group(s) of additive Digestibility enhancer (a) Composition, description Preparation of endo-1,4- beta-xylanase and endo-1,3(4)- beta-glucanase produced from Talaromyces versatilis IMI 378536 (basionym Penicillium funiculosum) and Talaromyces versatilis DSM 26702 (basionym Penicillium funiculosum) having a minimum activity of: Powder form - Endo-1,4-beta-xylanase: min. 22 000 viscometric units/g OR min. 2 300 DNS units/g - Endo-1,3(4)-beta-glucanase : min. 15 200 viscometric units/g OR min. 1 600 DNS units/g Liquid form - Endo-1,4-beta-xylanase: min. 5 500 viscometric units/ml OR min. 570 DNS units/ml - Endo-1,3(4)-beta-glucanase : min. 3 800 viscometric units/ml OR min. 400 DNS units/ml Chemical formula Description Purity criteria Method of analysis - Viscosimetric method: based on decrease in viscosity produced by action of Endo-1,4- xylanase on xylan containing substrate and action of Endo-1,3(4)-betaglucanase on glucan substrate. Colorimetric (DNS) method: based of the enzymatic hydrolysis of xylan containing substrate and glucan substrate respectively for Endo-1,4 xylanase and Endo-1,3(4)- beta-glucanase Trade name Name of the holder of authorisation Rovabio Spiky ADISSEO France S.A.S. Conditions of use Species or category of animal All major and minor poultry species (for fattening, reared for laying and breeding - Maximum Age Minimum content Maximum content Withdrawal period Units/kg of complete feedingstuffs Endo-1,4-beta-xylanase: 1 100 viscosimetric units Endo-1,3(4)-betaglucanase: 760 viscosimetric units - - EFSA Journal 2015; 13(5):4106 5
Other provisions and additional requirements for the labelling Specific conditions or restrictions for use Specific conditions or restrictions for handling Post-market monitoring Specific conditions for use in complementary feedingstuffs In the directions for use of the additive and premixture, indicate the storage temperature, storage life, and stability to pelleting Do not breath, avoid contact with skin and eye There is no need for specific requirements for a post-market monitoring plan other than those established by Regulations No 178/2002 and No 183/2005. ADISSEO has a full traceability system in place, including procedures for recalls. A free emergency contact number is printed in each label and also available in the Material Safety Data Sheet supplied to all customers. For use in feed rich in starch and non-starch polysaccharides (mainly beta-glucans and arabinoxylans) Marker residue Maximum Residue Limit (MRL) Species or category of animal Target tissue(s) or food products - - - - Maximum content in tissues EFSA Journal 2015; 13(5):4106 6
ASSESSMENT 1. Introduction The additive Rovabio Spiky is a preparation of endo-1,4-beta-xylanase (xylanase: EC 3.2.1.8) and endo-1,3(4)-beta-glucanase (glucanase: EC 3.2.1.6) and is available in solid and liquid forms (Rovabio Spiky AP and Rovabio Spiky LC, respectively). This additive is prepared by combining two products of fermentation obtained from two strains of Penicillium funiculosum (Talaromyces versatilis sp. novus) (IMI 378536 and DSM 26702), one of which is non-genetically modified and the other is genetically modified. The formulations are considered equivalent in terms of safety and efficacy for the target species. The Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) issued an opinion on the safety and efficacy of Rovabio Spiky (endo-1,4-beta-xylanase and endo-1,3(4)-betaglucanase) as a feed additive for chickens for fattening, chickens reared for laying and other minor poultry species (for fattening and reared for laying) (EFSA FEEDAP Panel, 2014). This opinion considered the safety aspects of the additive regarding the consumer, the user, the environment and the genetic modification of one of the production strains. The applicant seeks the authorisation of the additive as a feed additive for turkeys for fattening, turkeys reared for breeding and minor poultry species for fattening, reared for laying and reared for breeding at a minimum recommended dose of 1 100 U xylanase and 760 U glucanase per kg feed. In support of this extension, studies on turkeys for fattening as a second major poultry species are submitted. The Panel considers that aspects other than the safety and efficacy for turkeys have been addressed in the previous opinion and is not aware of any new information that would lead it to reconsider the previous conclusions. Therefore, the present assessment will address these aspects for the new target species only. 2. Conditions of use Rovabio Spiky is proposed for use in turkeys for fattening, turkeys reared for breeding and minor poultry species for fattening, reared for laying or reared for breeding at a minimum recommended dose of 1 100 U xylanase and 760 U glucanase per kg feed. 3. Evaluation of the analytical methods by the European Union Reference Laboratory (EURL) The EURL considered that the conclusions and recommendations reached in the previous assessment are valid and applicable for the current application. 7 4. Safety for the target species A total of 900 one-day-old male and female chicks (Hybrid Grade Maker, commercial strain) were distributed in groups of 25 birds (sex separated) in 36 pens and were allocated to one of three dietary treatments (six male and six female replicates per treatment). 8 Starter and grower diets based on wheat, soya bean meal and wheat distillers dried grain with solubles (DDGS) were supplemented with Rovabio Spiky AP to provide (xylanase/glucanase) 0/0, 2 200/1 520 (2 minimum recommended dose) or 220 000/152 000 (200 minimum recommended dose) units per kg feed. The enzyme activities (xylanase/glucanase) were confirmed by analysis (1 936/1 431 for 2 and 217 014/167 244 for 200 minimum recommended dose). The feed was offered ad libitum for 42 days as crumble (first 21 days) or as pellets (from day 22 to day 42). Feed consumption and body weight of the birds were 7 The full report is available on the EURL website: https://ec.europa.eu/jrc/sites/default/files/finrep-fad-2013-0030- RovabioSpiky.pdf 8 Technical dossier/section III/Annex III.1.1. EFSA Journal 2015; 13(5):4106 7
measured on days 21 and 42 of the study. An analysis of variance (ANOVA) was performed on the data obtained (including treatment, sex, block and the interaction between treatment and sex as effects). The pen was considered the experimental unit for all measured parameters. The mortality data were analysed using a chi-squared test. No interaction between the dietary treatment and sex were registered. Mortality was low (< 2.0 %) and was not affected by the dietary treatments. Supplementation of the experimental diets with 200-fold the minimum recommended dose did not have adverse effects on the performance of the birds. Mean final body weight of the birds (mean of male and female) was 2.0 kg per bird in the control diet and 2.1 kg per bird in the supplemented diets (P < 0.01). Total feed intake was not affected by the dietary treatments and was close to 3 450 g per bird. The feed-to-gain ratio was improved (P < 0.001) by the enzyme supplementation at all doses (1.71, 1.67 and 1.63 for the 0/0, 2 and 200 doses, respectively). Based on the results obtained in the tolerance trial, the FEEDAP Panel concludes that the additive is safe for turkeys for fattening under the conditions of use proposed by the applicant (1 100 U/kg xylanase and 760 U/kg glucanase). This conclusion is extended to turkeys reared for breeding. The FEEDAP Panel considers that the results of the tolerance study in turkeys for fattening support the previous conclusion (EFSA FEEDAP Panel, 2014) that the additive is safe for minor poultry species for fattening, reared for laying and reared for breeding at the same dose. 5. Efficacy The applicant provided four long-term trials. One of these trials was not considered further because of its short duration (42 days). The three studies share a common design in which one-day-old turkeys (mixed sexes in trials 1 and 2 and females in trial 3) were randomly allocated to two dietary treatments (control or treated). Starter and grower diets based on wheat, maize, maize DDGS and soya bean meal (trials 1 and 2) or wheat and soya bean meal (trial 3) were supplemented with Rovabio Spiky LC (sprayed on the pellets) to provide (xylanase/glucanase) 0/0 or 1 100/760 units per kg feed. The enzyme activities were confirmed by analysis. The feed was offered to the birds ad libitum for 84 days as crumble (until day 21) or as pellets (the rest of the study). Diets contained monensin until week 9. Feed consumption and body weight were measured in each pen on days 21, 42, 63 and 84 of the study. ANOVA was performed on the data obtained for each of the three studies. A significant improvement (P < 0.05) in the feed-to-gain ratio of turkeys for fattening was observed in all three trials without a significant effect in feed intake and weight gain (Table 2). Table 2: Experimental design and results of the performance trials in turkeys for fattening Trial no Sex Total number of birds (replicates/treatment birds/replicate) Intended dose xylanase/glucanase (U/kg)* Total feed intake (g/bird) Weight gain (g/bird) Feedto-gain ratio Mortality (%) 1 600 (24 25) 0/0 15 895 7 243 2.17 a 0.7 1 100/760 15 562 7 259 2.13 b 0.3 2 1200 (48 25) 0/0 19 031 8 971 2.11 a 1.6 1 100/760 18 737 8 910 2.09 b 0.8 3 600 (24 25) 0/0 13 938 6 443 2.16 a 6.3 1 100/760 13 783 6 474 2.03 b 3.3 *: Analysed enzyme activities: trial 1: 1 073/776; trial 2: 1 260/858; trial 3: 1 235/851. a,b: Means with different superscripts within a trial indicate significant differences between treatment groups (P < 0.05). EFSA Journal 2015; 13(5):4106 8
The supplementation of the diets with Rovabio Spiky at the recommended dose resulted in a statistically significant improvement in the feed-to-gain ratio of turkeys for fattening in all three trials. The FEEDAP Panel concludes that the additive has the potential to be efficacious in turkeys for fattening at the minimum recommended dose (1 100 U/kg xylanase and 760 U/kg glucanase). This conclusion is extended to turkeys reared for breeding. The FEEDAP Panel considers that the results of the efficacy studies in turkeys support the previous conclusion (EFSA FEEDAP Panel, 2014) that the additive has the potential to be efficacious in minor poultry species for fattening, reared for laying and reared for breeding at the same dose. 6. 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 9 and Good Manufacturing Practice. CONCLUSIONS The solid and liquid formulations are considered equivalent in terms of safety and efficacy for the target species. The additive is safe and has the potential to be efficacious in turkeys for fattening at the minimum recommended dose (1 100 U/kg xylanase and 760 U/kg glucanase). This conclusion is extended to turkeys reared for breeding. Moreover, the Panel confirms the previous conclusions that the additive is safe and efficacious in minor poultry species for fattening, reared for laying and reared for breeding at the same conditions of use. DOCUMENTATION PROVIDED TO EFSA 1. Rovabio Spiky for turkeys for fattening and other minor poultry species (for fattening and reared for laying or breeding). May 2014. Submitted by ADISSEO France SAS. 2. Rovabio Spiky for turkeys for fattening and other minor poultry species (for fattening and reared for laying or breeding). Supplementary information. December 2014. Submitted by ADISSEO France SAS. 3. Comments from Member States received through the ScienceNet. REFERENCES EFSA FEEDAP Panel (EFSA Panel on Additives and Products or Substances used in Animal Feed), 2014. Scientific Opinion on the safety and efficacy of Rovabio Spiky (endo-1,4-beta-xylanase and endo-1,3(4)-beta-glucanase) as a feed additive for chickens for fattening, chickens reared for laying and other minor poultry species (for fattening and reared for laying). EFSA Journal 2014;12(7):3793, 20 pp. doi:10.2903/j.efsa.2014.3793 9 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. EFSA Journal 2015; 13(5):4106 9