Safety and efficacy of fumonisin esterase from Komagataella phaffii DSM 32159 as a technological feed additive for pigs and poultry
2018; Wiley; Volume: 16; Issue: 5 Linguagem: Inglês
10.2903/j.efsa.2018.5269
ISSN1831-4732
AutoresGuido Rychen, Gabriele Aquilina, Giovanna Azimonti, Vasileios Bampidis, Maria de Lourdes Bastos, Georges Bories, Andrew Chesson, Pier Sandro Cocconcelli, Gerhard Flachowsky, Jürgen Gropp, Boris Kolar, Maryline Kouba, Marta López‐Alonso, Secundino López Puente, Baltasar Mayo, Fernando Ramos, Maria Saarela, Roberto Edoardo Villa, Robert John Wallace, Pieter Wester, Giovanna Martelli, Derek Renshaw, Boet Glandorf, Lieve Herman, Sirpa Kärenlampi, Jaime Aguilera, Gloria López‐Gálvez, Alberto Mantovani,
Tópico(s)Protein Hydrolysis and Bioactive Peptides
ResumoEFSA JournalVolume 16, Issue 5 e05269 Scientific OpinionOpen Access Safety and efficacy of fumonisin esterase from Komagataella phaffii DSM 32159 as a technological feed additive for pigs and poultry EFSA Panel on Additives and Products or Substances used in Animal Feed (EFSA FEEDAP Panel), Search for more papers by this authorGuido Rychen, Search for more papers by this authorGabriele Aquilina, Search for more papers by this authorGiovanna Azimonti, Search for more papers by this authorVasileios Bampidis, Search for more papers by this authorMaria de Lourdes Bastos, Search for more papers by this authorGeorges Bories, Search for more papers by this authorAndrew Chesson, Search for more papers by this authorPier Sandro Cocconcelli, Search for more papers by this authorGerhard Flachowsky, Search for more papers by this authorJürgen Gropp, Search for more papers by this authorBoris Kolar, Search for more papers by this authorMaryline Kouba, Search for more papers by this authorMarta López-Alonso, Search for more papers by this authorSecundino López Puente, Search for more papers by this authorBaltasar Mayo, Search for more papers by this authorFernando Ramos, Search for more papers by this authorMaria Saarela, Search for more papers by this authorRoberto Edoardo Villa, Search for more papers by this authorRobert John Wallace, Search for more papers by this authorPieter Wester, Search for more papers by this authorGiovanna Martelli, Search for more papers by this authorDerek Renshaw, Search for more papers by this authorBoet Glandorf, Search for more papers by this authorLieve Herman, Search for more papers by this authorSirpa Kärenlampi, Search for more papers by this authorJaime Aguilera, Search for more papers by this authorGloria López-Gálvez, Search for more papers by this authorAlberto Mantovani, Search for more papers by this author EFSA Panel on Additives and Products or Substances used in Animal Feed (EFSA FEEDAP Panel), Search for more papers by this authorGuido Rychen, Search for more papers by this authorGabriele Aquilina, Search for more papers by this authorGiovanna Azimonti, Search for more papers by this authorVasileios Bampidis, Search for more papers by this authorMaria de Lourdes Bastos, Search for more papers by this authorGeorges Bories, Search for more papers by this authorAndrew Chesson, Search for more papers by this authorPier Sandro Cocconcelli, Search for more papers by this authorGerhard Flachowsky, Search for more papers by this authorJürgen Gropp, Search for more papers by this authorBoris Kolar, Search for more papers by this authorMaryline Kouba, Search for more papers by this authorMarta López-Alonso, Search for more papers by this authorSecundino López Puente, Search for more papers by this authorBaltasar Mayo, Search for more papers by this authorFernando Ramos, Search for more papers by this authorMaria Saarela, Search for more papers by this authorRoberto Edoardo Villa, Search for more papers by this authorRobert John Wallace, Search for more papers by this authorPieter Wester, Search for more papers by this authorGiovanna Martelli, Search for more papers by this authorDerek Renshaw, Search for more papers by this authorBoet Glandorf, Search for more papers by this authorLieve Herman, Search for more papers by this authorSirpa Kärenlampi, Search for more papers by this authorJaime Aguilera, Search for more papers by this authorGloria López-Gálvez, Search for more papers by this authorAlberto Mantovani, Search for more papers by this author First published: 17 May 2018 https://doi.org/10.2903/j.efsa.2018.5269Citations: 3 Correspondence: feedap@efsa.europa.eu Requestor: European Commission Question number: EFSA-Q-2017-00073 Panel members: Gabriele Aquilina, Giovanna Azimonti, Vasileios Bampidis, Maria de Lourdes Bastos, Georges Bories, Andrew Chesson, Pier Sandro Cocconcelli, Gerhard Flachowsky, Jürgen Gropp, Boris Kolar, Maryline Kouba, Marta López-Alonso, Secundino López Puente, Alberto Mantovani, Baltasar Mayo, Fernando Ramos, Guido Rychen, Maria Saarela, Roberto Edoardo Villa, Robert John Wallace and Pieter Wester. Legal notice: Relevant information or parts of this scientific output have been blackened in accordance with the European Commission decision on the confidentiality requests formulated by the applicant pending. A previous, provisional version of this output which had been made publicly available pending the adoption of the decision has been replaced by this version. The full output has been shared with the European Commission, European Union (EU) Member States and the applicant. Acknowledgements: The FEEDAP Panel wishes to thank the following for the support provided to this scientific output (in alphabetical order of the last name): Jaume Galobart, Lucilla Gregoretti and Paola Manini. Adopted: 17 April 2018 Amended: 5 June 2018 AboutSectionsPDF ToolsExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onEmailFacebookTwitterLinked InRedditWechat Abstract Fumonisin esterase produced from a genetically modified strain of Komagataella phaffi is intended to degrade fumonisin mycotoxins contaminants in feeds for pigs and poultry. The production strain and its recombinant genes are not present in the final product. The applicant selected 300 U/kg feed to represent a likely upper limit. This concentration showed to be safe for piglets, chickens and turkeys for fattening and laying hens; the additive is thus safe for those categories. This conclusion is extended to all pigs, chickens reared for laying and turkeys reared for breeding and extrapolated to all other poultry species for growing and laying and to minor porcine species. No evidence of mutagenicity or genotoxicity was detected and no evidence of toxicity from a repeated-dose oral toxicity study; the residue assessment did not identify any concern. The use of the additive is, thus, considered safe for consumers. The additive is not toxic by inhalation and the respiratory exposure is likely to be low; however, a risk of sensitisation via the respiratory route cannot be excluded. The additive is non-irritant to skin and eyes and is not considered a dermal sensitiser. No risks for the environment are expected following the use of the additive in feeds under the proposed condition of use. The additive has the capacity to degrade fumonisin contaminants in feed of marketable quality when used at the minimum recommended dose of 10 U/kg complete feed, as shown in studies with chickens for fattening, laying hens and weaned piglets. Since the mode of action of the additive can be reasonably assumed to be the same in animal species for which the application is made, the Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) considers the additive efficacious for all poultry and all pigs. 1 Introduction 1.1 Background and Terms of Reference Regulation (EC) No 1831/200311 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. 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 Biomin GmbH22 Biomin GmbH, Erber Campus 1, 3131 Getzersdorf, Austria. for authorisation of the product fumonisin esterase (FUMzyme®), when used as a feed additive for all pigs and all avian species (category: technological additives; functional group: substances for reduction of the contamination of feed by mycotoxins). 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. The particulars and documents in support of the application were considered valid by EFSA as of 2 March 2017. 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 fumonisin esterase (FUMzyme®), when used under the proposed conditions of use (see Section 3.1.7). 1.2 Additional information The product under assessment is an enzyme-based additive intended to degrade fumonisin mycotoxins found as contaminants of feed. The safety and efficacy of the enzyme expressed in a different host from that used in the present application were the subjects of two previous opinions: the first, published in 2014, considered its use in feeds for pigs (EFSA FEEDAP Panel, 2014) and the second in 2016 its use in feed for all avian species (EFSA FEEDAP Panel, 2016). This additive is currently authorised for use in pigs33 Commission Implementing Regulation (EU) No 1115/2014 of 21 October 2014 concerning the authorisation of a preparation of fumonisin esterase produced by Komagataella pastoris (DSM 26643) as a feed additive for pigs. OJ L 302, 22.10.2014, p. 51. and avian species (1m03).44 Commission Implementing Regulation (EU) 2017/913 of 29 May 2017 concerning the authorisation of a preparation of fumonisin esterase produced by Komagataella pastoris (DSM 26643) as a feed additive for all avian species. OJ L 139, 30.5.2017, p. 33. 2 Data and methodologies 2.1 Data The present assessment is based on data submitted by the applicant in the form of a technical dossier55 FEED dossier reference: FAD-2017-0005. in support of the authorisation request for the use of FUMzyme® as a feed additive. The technical dossier was prepared following the provisions of Article 7 of Regulation (EC) No 1831/2003, Regulation (EC) No 429/200866 Commission Regulation (EC) No 429/2008 of 25 April 2008 on detailed rules for the implementation of Regulation (EC) No 1831/2003 of the European Parliament and of the Council as regards the preparation and the presentation of applications and the assessment and the authorisation of feed additives. OJ L 133, 22.5.2008, p. 1. and the applicable EFSA guidance documents. The FEEDAP Panel used the data provided by the applicant together with data from other sources, such as previous risk assessments by EFSA or other expert bodies. EFSA has verified the European Union Reference Laboratory (EURL) report as it relates to the methods used for the control of the fumonisin esterase (EC 3.1.1.87) (FUMzyme®) in animal feed. The Executive Summary of the EURL report can be found in Annex A.77 The full report is available on the EURL website: https://ec.europa.eu/jrc/sites/jrcsh/files/finrep_fad-2017-0005_fumzyme.pdf 2.2 Methodologies The approach followed by the FEEDAP Panel to assess the safety and the efficacy of FUMzyme® is in line with the principles laid down in Regulation (EC) No 429/2008 and the relevant guidance documents: Guidance on technological additives (EFSA FEEDAP Panel, 2012a), Technical guidance: Tolerance and efficacy studies in target animals (EFSA FEEDAP Panel, 2011), Technical Guidance for assessing the safety of feed additives for the environment (EFSA, 2008), Guidance for establishing the safety of additives for the consumer (EFSA FEEDAP Panel, 2012b), Guidance on studies concerning the safety of use of the additive for users/workers (EFSA FEEDAP Panel, 2012c). 3 Assessment The present application concerns an additive containing the enzyme fumonisin esterase as the single active agent intended for use as a technological additive (functional group substances for reduction of the contamination of feed by mycotoxins). 3.1 Characterisation 3.1.1 Characterisation of the active substance The active substance, the enzyme fumonisin esterase (EC 3.1.1.87), partially degrades fumonisin B1 (FB1) and related fumonisins by cleavage of the diester bonds and release of the tricarballylic acid and is intended to reduce the toxicity of contaminated feed. The enzyme was identified in a Sphingopyxis sp., an α-Proteobacterium isolated from soil, and the genes encoding the activity used to transform a strain of the yeast Komagataella phaffii (formerly Pichia pastoris) which is then used as the production organism.88 Technical Dossier/Section II/Annex II.19. 3.1.2 Characterisation of production organism The additive is produced with a genetically modified strain of K. phaffii, which is deposited in the German Collection of Microorganisms and Cell Cultures as P. pastoris with deposit number DSM 32159.■■■■■ The strain was identified as K. phaffii by sequence analysis of the 26S/28S and ITS regions of the rRNA gene, and partial sequence of the EF-1α gene.1010 Technical Dossier/Supplementary Information/Annex (i). 3.1.2.1 Characteristics of the recipient or parental microorganism The recipient strain is a genetically modified derivative of the parental strain K. phaffii CBS 7435 (NRRL Y-11430, ATCC 76273). The whole-genome sequence of this strain is published (Küberl et al., 2011). The strain was originally taxonomically classified as P. pastoris, a species which is considered by EFSA to be suitable for the Qualified Presumption of Safety (QPS) approach to safety assessment (EFSA BIOHAZ Panel, 2017) when used for enzyme production. Recently, this strain was renamed as K. phaffii by molecular taxonomy analyses. The FEEDAP Panel considers that this reclassification does not have an impact in the safety status of the strain. ■■■■■ ■■■■■■■■■■11 ■■■■■ 3.1.2.2 Characteristics of the donor organism The fumD ■■■■■ gene, coding for a FB1 carboxylesterase, is a codon-optimised synthetic sequence based on the fumD gene of the α-proteobacterium Sphingopyxis sp. MTA144.1212 Technical Dossier/Section II/Annex II.27. This strain, which was isolated from soil, carries a gene cluster associated with fumonisin degradation as described by Heinl et al. (2010). The gene fumD codes for a carboxylesterase, which catalyses the hydrolysis of fumonisin B1 to aminopentol 1 (producing fully de-esterified fumonisin, HFB1). ■■■■■ 3.1.2.3 Description of the genetic modification ■■■■■■■■■■13 ■■■■■ ■■■■■ ■■■■■■■■■■14 ■■■■■ ■■■■■ ■■■■■■■■■■14 ■■■■■ ■■■■■■■■■■15 ■■■■■ ■■■■■■■■■■16 ■■■■■ ■■■■■ 3.1.3 Manufacturing process Fumonisin esterase is produced by the submerged fermentation of the production strain. After fermentation, the biomass is separated by centrifugation, and the supernatant is then micro- and ultrafiltered. The final additive is prepared from the high-molecular weight material recovered from ultrafiltration mixed with 10% maltodextrin, which is then spray dried. The dried material is then blended with maltodextrin. FUMzyme® is routinely quality controlled for the presence/absence of cultivable K. phaffii, following a specific procedure.1717 Technical dossier/Section II/Annex II.65. The absence of viable cells of the production strain is tested in 25 g after a resuscitation step in non-selective liquid medium at 37°C for 6–8 h, followed by plating 0.1 mL of the suspension onto selective medium and incubating at 30°C for 48–96 h.1818 Technical dossier/Section II/Annex II.66. No viable cells detected in three batches analysed in triplicate.2020 ■■■■■ Recombinant DNA is also removed during downstream processing: no recombinant DNA was detected in 100 mg of three batches of the final product by real-time PCR, ■■■■■■■■■■20 ■■■■■ 3.1.4 Characterisation of the additive The additive typically contains approximately 8% of material derived from the fermentation and 90% of the added maltodextrin carrier and has a specified minimum content of 3,000 U esterase/g. One unit (U) is the enzymatic activity that releases 1 μmol propane-1,2,3-tricarboxylic acid per minute from 100 μM FB1 in 20 mM Tris-Cl buffer pH 8.0 with 0.1 mg/mL bovine serum albumin at 30°C. Compliance with this minimum specification was demonstrated for five batches of the additive (mean 3,470 U/g, range 3,150–3,780 U/g additive).2121 Technical Dossier/Section II/Annex II.03. Routine monitoring for the presence of heavy metals and arsenic (As) in three different production batches of the additive showed that the concentrations of lead (Pb), cadmium (Cd), mercury (Hg) and As were in all cases below the respective limits of quantification (Pb < 0.5, Cd < 0.1, Hg < 0.01 and As < 0.5 mg/kg).2222 Technical Dossier/Section II/Annexes II.03, II.05 and II.06. Quality control parameters for microbial contaminants are set at a maximum of 30 colony-forming units (CFU)/g for total coliforms, a maximum of 100 CFU/g individually for yeasts and filamentous fungi and the absence of Escherichia coli and Salmonella spp. in 25 g product; certificates of analysis for three batches of product show compliance with these values.2323 Technical Dossier/Section II/Annexes II.08, II.09 and II.10. A further three batches were examined for the presence of 11 mycotoxins commonly associated with cereals2424 Technical Dossier/Section II/Annex II.07. ; all values were below the limits of detection.2525 Limits of detection (in µg/kg) are listed here. Deoxynivalenol: 20, zearalenone: 4, aflatoxin B1/B2/G1: 0.2, fumonisin B1/B2: 20, HT-2 toxin/T-2 toxin: 2. Particle size analysis by laser diffraction (three batches) showed that approximately 60% of the final product had a particle size < 100 μm, 40% < 50 μm and ~ 5% of respirable size (< 10 μm).2626 Technical Dossier/Section II/Annex II.12. The dusting potential of the same three batches determined according to the Stauber–Heubach method gave a mean value of 0.25 g/m3.2727 Technical Dossier/Section II/Annexes II.13, II.14 and II.15. 3.1.5 Stability and homogeneity 3.1.5.1 Shelf-life The shelf-life of the product was established using five batches of product stored at room temperature or at 37°C for 18 months in sealed containers.2828 Technical Dossier/Section II/Annex II.67. Samples were analysed for activity at six monthly intervals. There was essentially no loss of activity at room temperature after 18 months and only a small loss at the higher temperature (10–15%). 3.1.5.2 Stability in premixes and feed A single batch of the additive was mixed with a commercial bentonite mineral feed at 20,000 U/kg and a commercial vitamin–mineral premix (containing choline chloride) to a concentration of 15,000 U/kg.2929 Technical Dossier/Section II/Annex II.68. The samples were then stored for 6 months at room temperature. No loss of activity of any significance was seen after 6 months storage under these conditions. Four batches of feed (in mash form) containing the additive were prepared, the first three based on a grower diet for pigs supplemented with 10, 15 or 20 U/kg feed and the fourth on a feed for chickens supplemented with 45 U/kg feed.3030 Technical Dossier/Section II/Annex II.69. Activity was measured at the time of preparation and again after 6 months storage at 22 ± 2°C. Essentially no loss of activity was recorded. 3.1.5.3 Homogeneity A single batch of additive was mixed with additional maltodextrin and then mixed with 99 kg piglet feed using a farm-scale feed mill.3131 Technical Dossier/Section II/Annex II.70. The treated feed with an intended concentration of 450 U/kg feed was then distributed equally to 10 bags and, from each bag, 10 subsamples were taken and analysed for activity. The results showed a mean value of 477 U/kg feed with a coefficient of variation (CV) of 3.6%. 3.1.6 Interference with the analysis of mycotoxins in feed The possible interference of the additive in the determination of fumonisins in feedstuffs was examined.3232 Technical Dossier/Section II/Annex II.71. The methanol/acetonitrile/water solution used for the initial extraction of fumonisins in the standard method was found to fully inhibit the action of fumonisin esterase and, therefore, no interference is expected. Given the specificity of the esterase, the analytical determination of other structural classes of mycotoxins would not be affected. 3.1.7 Conditions of use The additive is intended for use with pigs and poultry at a minimum inclusion rate of 10 U/kg complete feed. No maximum incorporation rate is proposed. No withdrawal period is foreseen. 3.2 Safety 3.2.1 Safety aspects of the genetic modification The original strain from which the production organism was derived belong to K. phaffii, formerly identified as P. pastoris, a species which is considered by EFSA to be suitable for the QPS approach to safety assessment when used for enzyme production (EFSA BIOHAZ Panel, 2017). The FEEDAP Panel considers that this reclassification does not have an impact in the safety status of the strain. The production strain carries several copies of the Zeocin™ resistance gene and the Geneticin™ resistance gene integrated into its chromosome. Neither the viable production strain nor its recombinant DNA was found in the final product. Therefore, the product FUMzyme®, manufactured by fermentation with K. phaffii DSM 32159, does not give rise to safety concerns deriving from the genetic modification of the production strain. 3.2.2 Toxicological studies The toxicity studies were conducted with a non-standardised production batch in which the proportion of the culture supernatant was greater than the standardised product resulting in higher concentrations of the enzyme (8,650 U/g). 3.2.2.1 Genotoxicity Bacterial reverse mutation test Two independent experiments were made in compliance with the Organisation for Economic Co-operation and Development (OECD) guideline 471, with the Salmonella Typhimurium strains TA1535, TA1537, TA98, TA100 and TA102 with and without metabolic activation.3333 Technical Dossier/Section III/Annex III.34. Six doses were tested, starting at 31.6 μg and increasing to a top dose of 5,000 μg. No precipitation of the test item or toxic effects was seen. No increase in the number of revertant colonies was seen at any dose tested in the presence or absence of metabolic activation. The additive is, therefore, considered non-mutagenic under the conditions of the assay. In vitro chromosome aberration test This test was made according to the OECD guideline 473 using the Chinese hamster V79 cell line.3434 Technical Dossier/Section III/Annex III.35. Chromosomes were prepared 21 h after the start of treatment with the test item and 150 metaphases per culture scored for structural chromosome aberrations. Treatment intervals were 4 h with and without metabolic activation in experiment 1 and 4 h with activation and 20 h without activation in experiment 2. The doses tested were 1,000, 1,500 and 2,000 μg/mL in both parts of the test. The appropriate positive controls were included (ethyl-methanesulfonate and cyclophosphamide) which behaved as expected. The test item did not induce structural chromosome aberrations in either experiment and no increase in the frequency of polyploid cells was observed in comparison with the negative control. The test item is thus considered non-clastogenic under the conditions of the test. In vivo mammalian erythrocyte micronucleus test The test was made following OECD guideline 474 using mouse peripheral blood cells.3535 Technical Dossier/Section III/Annex III.36. Blood samples were collected for analysis from 10 mice (strain NMRI; five males and five females) per treatment 44 and 66 h after a single administration of the test item. A maximum tolerated dose (MTD) of 2,000 mg/kg body weight (bw) was selected on the basis of a preliminary range finding experiment with intermediate doses of 1,000 and 400 mg/kg bw, representing 0.5 and 0.2 of the MTD. Animals given the two lower doses showed no clinical symptoms. Those given the top dose showed slight signs of systemic toxicity. For all doses, 10,000 polychromatic erythrocytes per animal were scored for the incidence of micronucleated immature cells. Negative controls were within the historical data and the positive control (cyclophosphamide) behaved as expected. No biologically relevant increase in micronucleated cells was seen at any dose, from which it was concluded that the test item is non-mutagenic with respect to clastogenicity and/or aneugenicity. 3.2.2.2 Repeated -dose 90-day oral toxicity study A Good Laboratory Practice (GLP) compliant repeat dose study was made with equal numbers of male and female Wistar rats following the protocol for OECD guideline 408.3636 Technical Dossier/Section III/Annex III.37. Animals were caged in groups of five animals of the same sex. Twenty animals (two cages of each sex per treatment) were assigned to each of a vehicular control and groups given daily 100, 1,000 and 2,000 mg/kg bw of the test item via the feed for 90 days. A further satellite group of 10 animals was also given the high dose for 90 days. Body weight was determined at the start and weekly thereafter and body condition monitored daily. Blood samples were taken on day 90 from the control and high-dose groups for haematology and clinical chemistry. A complete necropsy of all animals was made with histopathology for tissues from the high-dose group. Animals in the satellite group were observed for a further 28 post-treatment days before necropsy on day 118. No mortalities, behavioural changes or adverse clinical signs were noted during the study. Growth of males and females was not significantly different between the treatment groups. No statistically significant changes in clinical chemistry results were seen in either sex or in haematology results for males. The only significant difference observed was a change in the relative volume of lymphocytes in female rats given the top dose compared to the control group. This change is reported as being within the historical range for the colony. There were no significant differences in relative or absolute organ weights in male rats, but significant differences were seen in the absolute weight of uterus and ovaries and relative weight of ovaries. These changes were not dose related and were within historical values and were attributed by the study authors to animals at different stages of their oestrus cycle. 3.2.2.3 Metabolites resulting from the degradation of fumonisins Metabolites resulting from the complete or partial de-esterification of fumonisins were assessed for safety in the previous opinion on the use of a fumonisin esterase in pigs and found to be less toxic than the parent mycotoxin (EFSA FEEDAP Panel, 2014). Since the action of the esterase on any contaminating fumonisins will be essentially the same in the pig and poultry digestive tracts and independent of the source of the enzyme, the previous assessment and its conclusion apply equally to the present application. 3.2.3 Safety for the target species The recipient strain K. phaffii used for the production of the enzyme is a species considered by EFSA to be suitable for the QPS approach to the assessment of safety. Since the genetic modifications give no cause for concern and since the enzyme itself has been assessed for safety in the context of previous opinions, there is little reason to consider further target animal safety. However, the applicant has chosen to provide four additional tolerance studies with weaned piglets, chickens and turkeys for fattening and laying hens. 3.2.3.1 Safety for weaned piglets A total of 96 weaned piglets (mixed sex O-HYB-F1 with mean start weight of approximately 10 kg) were assigned to one of three groups: a control group and two test gro
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