Assessment of the application for renewal of authorisation of PHYZYME®XP 5000 G/L (6-phytase) for chickens for fattening, laying hens, turkeys for fattening, ducks for fattening, weaned piglets, pigs for fattening and sows for reproduction.

PHYZYME ® XP 5000 G/L is a feed additive that contains 6-phytase produced by a genetically modified strain of Schizosaccharomyces pombe. The applicant requested for the renewal of the authorisation for PHYZYME ® XP 5000 G and L to be used as a feed additive in chickens for fattening, laying hens, turkeys for fattening, ducks for fattening, piglets (weaned), pigs for fattening and sows. This scientific opinion concerns the renewal of the authorisation of the additive for those species. To support the request, the applicant provided evidence that the additive in the market complies with the conditions of the authorisation. According to the information provided by the applicant, no new evidence has been identified that would make the FEEDAP Panel reconsider the previous conclusions regarding the safety for the target species, consumer, user and environment. The application for renewal of the authorisation did not include a proposal for amending or supplementing the conditions of the original authorisation that would have an impact on the efficacy of the additive. Therefore, there was no need for assessing the efficacy of the additive in the context of the renewal of the authorisation.

Introduction

Background and Terms of Reference

Regulation (EC) No 1831/20031 establishes the rules governing the Community authorisation of additives for use in animal nutrition. In particular, Article 14(1) of that Regulation lays down that an application for renewal shall be sent to the Commission at the latest one year before the expiry date of the authorisation.

The European Commission received a request from Danisco Animal Nutrition2 for renewal of the authorisation of the product PHYZYME® XP 5000 G/L (6‐phytase), when used as a feed additive for chickens, turkeys and ducks for fattening, laying hens, weaned piglets, pigs for fattening and sows (category: zootechnical additives; functional group: digestibility enhancers).

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 14(1) (renewal of the authorisation). The particulars and documents in support of the application were considered valid by EFSA as of 24 October 2016.

According to Article 8 of Regulation (EC) No 1831/2003, 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. EFSA shall deliver an opinion on the safety for the target animals, consumer, user and the environment and on the efficacy of the product PHYZYME® XP 5000 G/L (6‐phytase), when used under the proposed conditions of use (see Section 3.2.1).

Additional information

The additive PHYZYME® XP 5000 G/L is a preparation of 6‐phytase produced by a genetically modified strain of Schizosaccharomyces pombe (ATCC 5233). EFSA issued two opinions which considered the safety and efficacy of the enzyme preparation when used in chickens for fattening, laying hens, turkeys for fattening, ducks for fattening, piglets (weaned), pigs for fattening and sows, which included the assessment of the safety for the consumer, the user and the environment, as well as the safety aspects of the genetic modification (EFSA, 2006a,b). A third opinion was adopted regarding the use of a more concentrated form, PHYZYME®XP 10000 G and L (EFSA, 2008), and a fourth opinion on the extension of use of the additive for minor poultry species was adopted in 2012 (EFSA FEEDAP Panel, 2012a).

PHYZYME® XP 5000 XP G/L is authorised for use in chickens for fattening, laying hens, turkeys for fattening, ducks for fattening, piglets (weaned), pigs for fattening and sows3 , 4 and all avian species.5

The applicant requested the renewal of the authorisation for PHYZYME® XP 5000 G and L to be used as a feed additive in chickens for fattening, laying hens, turkeys for fattening, ducks for fattening, piglets (weaned), pigs for fattening and sows.

Data and methodologies

Data

The present assessment is based on data submitted by the applicant in the form of a technical dossier6 in support of the authorisation request for the use of PHYZYME® XP 5000 G/L as a feed additive.

The European Union Reference Laboratory (EURL) considered that the conclusions and recommendations reached in the previous assessment regarding the methods used for the control of the active substance are valid and applicable for the current application.7

Methodologies

The approach followed by the FEEDAP Panel to assess the safety and the efficacy of PHYZYME® XP 5000 G/L is in line with the principles laid down in Regulation (EC) No 429/20088 and the relevant guidance documents: Guidance on the renewal of the authorisation of feed additives (EFSA FEEDAP Panel, 2013), Guidance on zootechnical additives (EFSA FEEDAP Panel, 2012a,b) and Guidance on the risk assessment of genetically modified microorganisms and their products intended for food and feed use (EFSA GMO Panel, 2011).

Assessment

The additive PHYZYME® XP 5000 G/L is a feed additive that contains 6‐phytase produced by a genetically modified strain of S. pombe (ATCC 5233). This additive is authorised for use in all avian species for fattening and laying, weaned piglets, pigs for fattening and sows. This opinion deals with the renewal of the authorisation of PHYZYME® XP 5000 G/L as a zootechnical additive (functional group of digestibility enhancers) for chickens for fattening, laying hens, turkeys for fattening, ducks for fattening, piglets (weaned), pigs for fattening and sows.

Characterisation of the additive

The additive is authorised in solid, PHYZYME® XP 5000 G, and liquid, PHYZYME® XP 5000 L, forms.

The information submitted regarding the manufacturing process lists a series of modifications applied during the last years to the fermentation and the enzyme recovery process.■■■■■■■■■■ The Panel considers that these modifications do not have an impact on the final product and data regarding the characterisation supports this conclusion. The applicant declared that no antibiotics are used during the manufacturing process.10

PHYZYME® XP 5000 G contains the phytase (solids 1.4–2.1%), calcium propionate (0.1–0.3%), citric acid (0.2–0.8%) and wheat flour (up to 100%). The composition of this formulation is basically the same as the one previously related with minor modifications on the amounts used (e.g. citric acid from 0.4–1.0% to 0.2–0.8%). PHYZYME® XP 5000 L contains the phytase (solids 1.5–2.3% in the final additive), sorbitol (2–8%), sodium chloride (11–17%), sodium citrate (0.5–1.0%), sodium benzoate (0.1–0.2%), potassium sorbate (0.07–0.11%) and water.

The two formulations ensure a minimum guaranteed enzyme activity of 5,000 FTU11/g. The batch‐to‐batch variation was studied in three recent batches of each formulation. The mean enzyme activity in the PHYZYME® XP 5000 G was 6,198 FTU/g (range 5,969–6,637 FTU/g) and in PHYZYME® XP 5000 L was 6,343 FTU/g (range 5,890–6,902).12

Purity specifications for the two formulations include chemical compounds lead (≤ 5 mg/kg), cadmium (≤ 0.5 mg/kg), mercury (≤ 0.1 mg/kg) and arsenic (≤ 2 mg/kg) as well as absence of coliform)per gram of product), and absence of Escherichia coli and Salmonella spp. (negative in 25 mL). Compliance with these specifications was demonstrated in at least three recent batches of each formulation.13 Purity data submitted included also information on mycotoxins; aflatoxin B1 (< 0.1 μg/kg for solid, < 1 μg/kg for the liquid), ochratoxin A (< 0.2 μg/kg for the solid and < 2 μg/kg for liquid), zearalenone (< 25 μg/kg), T2‐toxin (< 10 μg/kg only in the solid), fumonisins (< 600 μg/kg only in the liquid) and deoxynivalenol (< 330 μg/kg). Microbiological purity also included total viable cells (< 1.7 × 105 colony forming units (CFU)/g solid formulation, < 7 CFU/g for liquid formulation). No antimicrobial activity was found in three batches of either formulation.14

The 6‐phytase present in the additive is obtained by fermentation with a genetically modified strain of S. pombe which is deposited at the American Type Culture Collection under deposition number SD‐5233.15 The taxonomic classification of the strain as S. pombe was confirmed ■■■■■■■■■■■■■■■ The assessment of the genetic modification was performed in a previous opinion (EFSA, 2006a) and the Panel concluded that the genetic modification does not raise any safety concern. The production strain has not been subject to any further genetic modification.

The presence of viable cells of the production strain was investigated in three batches of the liquid formulation and three of the solid formulation (analysis of three samples for each batch in duplicate).17 The samples of the liquid formulation were plated (1 mL) in a proprietary in‐house medium appropriate for S. pombe and incubated at 30°C for 72 h. The samples of the solid formulation were first dissolved (approximately 1:10) and then 1 mL was plated in yeast extract glucose agar at 25°C for 120 h. No cells were detected. A positive control was included in the analysis.

The presence of recombinant DNA was analysed in triplicate in three batches of the liquid formulation.18 The primers targeted the recombinant phytase gene, with an amplicon size of 1,182 bp and the methodology included a lysis step. The analysis showed no amplification in the samples (while positive polymerase chain reaction (PCR) control gave amplification). The limit of detection in samples spiked with DNA was < 1 ng/mL. The results in the liquid formulation apply also to the solid formulation.

Conditions of use

The additive is authorised at a minimum recommended level of 150 FTU/kg feed for laying hens, 250 FTU/kg feed for chickens, turkeys and ducks for fattening, weaned piglets and pigs for fattening, and at 500 FTU/kg feed for sows. The applicant does not ask for a modification of these conditions of use.

Under the other provisions, it is stated that ‘in the directions for use of the additive and premixture, indicate the storage temperature, storage life, and stability to pelleting. For use in feed containing more than 0,23% phytin bound phosphorus. For piglets (weaned) up to 35 kg of body weight’.

Safety

Previous assessments

EFSA issued two opinions (EFSA, 2006a,b) which considered the safety and efficacy of the enzyme preparation when used in chickens for fattening, laying hens, turkeys for fattening, ducks for fattening, piglets (weaned), pigs for fattening and sows, which included the assessment of the safety for the consumer, the user and the environment, as well as the safety aspects of the genetic modification. The Panel concluded that the additive is safe for the target species at the recommended conditions of use, that the genetic modification of the production strain is of no concern and the use of the product as a feed additive would be of no concern for the consumers of products derived from animals fed with the additive, or for the environment. Concerns for the user were limited to its potential as a respiratory sensitiser.

Further evidence

In 2007, EFSA established the Qualified Presumption of Safety (QPS) approach to safety assessment for microorganisms (EFSA, 2007). The production strain of the additive belongs to a species, S. pombe, that is considered to qualify for the QPS approach to safety assessment (EFSA, 2007 and EFSA BIOHAZ Panel, 2017). The taxonomic classification of the strain has been unambiguously established and the genetic modification raised no concerns. Therefore, the fermentation product obtained from the production strain does not raise safety concerns for the consumer and the environment.

In line with the requirements established in the EFSA guidance on the renewal (EFSA FEEDAP Panel, 2013), the applicant performed a literature search in order to provide evidence that in the light of the current knowledge the additive remains safe under the approved conditions for target species, consumers, users and the environment. The literature search was conducted in July 2016 and covered the period from 2005 to 2016.19 Search terms used, and the strategy followed were reported. The search terms included several key words including those for the production strain (Schizosaccharomyces pombe and deposit number), words related to safety (e.g. safe* and toxic*, poison*) as well as the target species. The search was conducted using three databases, CAB Abstracts, Veterinary Science Database and Medline. The literature search retrieved 38 publications but in none of these publications adverse events or safety issues concerning the additive were reported (See Appendix A).

The applicant claims that no adverse effects have been reported in the framework of its global monitoring plan.20

Conclusions on the safety

Based on the above and the fact that the additive and the conditions of use for the species/categories for which the additive is authorised have not been modified, the Panel considers that there is no evidence to reconsider the conclusions reached in previous assessments. Therefore, the additive is safe for chickens, turkeys and ducks for fattening, laying hens, weaned piglets, pigs for fattening and sows, consumers of products from animals fed the additive and the environment but it is considered a potential respiratory sensitiser.

Efficacy

The present application for renewal of the authorisation does not include a proposal for amending or supplementing the conditions of the original authorisation that would have an impact on the efficacy of the additive. Therefore, there is no need for assessing the efficacy of the additive in the context of the renewal of the authorisation.

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 Regulation21 and Good Manufacturing Practice.

Conclusions

The additive currently in the market complies with the existing conditions of authorisation.

The FEEDAP Panel concludes that PHYZYME® 5000 G/L remains safe for chickens, turkeys and ducks for fattening, laying hens, weaned piglets, pigs for fattening and sows, consumers of products from animals fed the additive and the environment under the approved conditions of authorisation. The additive is a potential respiratory sensitiser.

There is no need for assessing the efficacy of the additive in the context of the renewal of the authorisation.

Documentation provided to EFSA/Chronology

Abbreviations

colony forming unit

European Union Reference Laboratory

EFSA Panel on Additives and Products or Substances used in Animal Feed

polymerase chain reaction

Qualified Presumption of Safety

Adeola O, 2010. Phosphorus equivalency value of an Escherichia coli phytase in the diets of White Pekin ducks. Poultry Science, 89, 1199–1206.

Bhuiyan MM, Islam AF and Iji PA, 2010. Response of broiler chickens to diets containing artificially dried high‐moisture maize supplemented with microbial enzymes. South African Journal of Animal Science, 40, 348–362.

Brana DV, Ellis M, Castaneda EO, Sands JS and Baker DH, 2006. Effect of a novel phytase on growth performance, bone ash, and mineral digestibility in nursery and grower‐finisher pigs. Journal of Animal Science, 84, 1839–1849.

Ciofalo V, Barton N, Kretz K, Baird J, Cook M and Shanahan D, 2003. Safety evaluation of a phytase, expressed in Schizosaccharomyces pombe, intended for use in animal feed. Regulatory Toxicology and Pharmacology, 37, 286–292.

Debicki‐Garnier AM, Sands J, Peron A and Messager B, 2007. Efficacy and mode of action of a new phytase in poultry feed. Actes des 7emes Journees de la Recherche Avicole, Tours, France, 28 et 29 mars 2007, 150–153.

EFSA FEEDAP Panel (EFSA Panel on Additives and Products or Substances used in Animal Feed), 2012. Scientific Opinion on the safety and efficacy of Phyzyme XP (6‐phytase) as a feed additive for minor poultry species. EFSA Journal 2012;10(3):2619, 9 pp. https://doi.org/10.2903/j.efsa.2012.2619

Ghazalah AA, Abd‐Elsamee MO, El‐Hakim ASA and Ibrahim MM, 2012. Evaluation of using distillers dried grains with solubles (DDGS) in broiler diets. Egyptian Poultry Science Journal, 32, 381–397.

Ghorbani MR, Fayazi J and Chaji M, 2009. Effect of dietary phytase and NSP‐degrading enzymes in diets containing rape seed meal on broiler performance and carcass characteristic. Research Journal of Biological Sciences, 4, 258–264.

Graham AB, DeRouchey JM, Tokach MD, Goodband RD, Dritz SS, Nitikanchana S, de Jong JA and Nelssen JL, 2012. The interactive effects of easyzyme and phytase in diets containing high‐fiber co‐products on growth performance of nursery pigs. Kansas State University Swine Day 2012. Report of Progress 1074. p. 66–73.

Guggenbuhl P, Quintana AP and Nunes CS, 2011. Comparative effects of two types of phytase added to a diet with reduced nutrient and energy content on the performance and digestibility of minerals in the weaned piglet. Journées de la Recherche Porcine en France, 43, 213–214.

Guggenbuhl P, Wache Y, Nunes CS and Fru F, 2012. Comparative effects of three phytases on the phosphorus and calcium use in the weaned piglet. Journal of Animal Science, 90, 95–97.

Humer E, Wetscherek W, Schwarz C and Schedle K, 2013. Effect of maize conservation technique and phytase supplementation on total tract apparent digestibility of phosphorus, calcium, ash, dry matter, organic matter and crude protein in growing pigs. Animal Feed Science and Technology, 185, 70–77.

Jones CK, Tokach MD, Ratliff BW, Horn NL, Dritz SS, Goodband RD, DeRouchey JM and Nelssen JL, 2009. Efficacy of different commercial phytase sources and development of a phosphorus release curve. Kansas State University Swine Day 2009. Report of Progress 1020. p. 106–121.

Jones CK, Tokach MD, Dritz SS, Ratliff BW, Horn NL, Goodband RD, DeRouchey JM, Sulabo RC and Nelssen JL, 2010. Efficacy of different commercial phytase enzymes and development of an available phosphorus release curve for Escherichia coli‐derived phytases in nursery pigs. Journal of Animal Science, 88, 3631–3644.

Joven M, Suárez‐Belloch J, Latorre MA and Fondevila M, 2015. Effect of addition of phytases on in vitro digestibility of ingredients for pig feeding, XVI Jornadas sobre Producción Animal, 19 y 20 de mayo de 2015, Zaragoza, España. Tomo I & II, 2015, p. 293–295.

Kerr BJ, Weber TE, Miller PS and Southern LL, 2010. Effect of phytase on apparent total tract digestibility of phosphorus in corn‐soybean meal diets fed to finishing pigs. Journal of Animal Science, 88, 238–247.

Khattak FM, Acamovic T and Cowieson AJ, 2009. The response of chicks, with prior exposure to low and high phytate diets, when subsequently fed diets supplemented with phytase and phytate. British Poultry Abstracts, 5, 35–36.

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Date	Event
01/08/2016	Dossier received by EFSA. PHYZYME® XP 5000 G/L Renewal. Submitted by Danisco Ltd.
12/09/2016	Reception mandate from the European Commission
24/10/2016	Application validated by EFSA – Start of the scientific assessment
07/04/2017	Request of supplementary information to the applicant in line with Article 8(1)(2) of Regulation (EC) No 1831/2003 – Scientific assessment suspended. Issues: characterisation
24/01/2017	Comments received from Member States
31/05/2018	Clarification teleconference during risk assessment with the applicant according to the “EFSA's Catalogue of support initiatives during the life‐cycle of applications for regulated products”
15/06/2018	Reception of supplementary information from the applicant ‐ Scientific assessment re‐started
29/06/2018	Request of supplementary information to the applicant in line with Article 8(1)(2) of Regulation (EC) No 1831/2003 – Scientific assessment suspended Issues: Characterisation
27/08/2018	Reception of supplementary information from the applicant ‐ Scientific assessment re‐started
04/04/2019	Opinion adopted by the FEEDAP Panel. End of the Scientific assessment