Abstract
Following a request from the European Commission, EFSA was asked to deliver a scientific opinion on the safety and the efficacy of the feed additives cellulase (produced by Aspergillus niger CBS 120604), beta‐glucanase (produced by Aspergillus neoniger MUCL 39199) and xylanases (produced by Trichoderma citrinoviride MUCL 39203 and CBS 614.94) as silage additives for all animal species. In 2018, the FEEDAP Panel evaluated 11 enzymes to be used as silage additives. In that opinion, the additives, including the production strains, were characterised in full and their safety and efficacy were evaluated. However, the Panel could not conclude on the identification of the production strains of the xylanase produced by the strain with deposit number MUCL 39203 (product referred as to J) and the glucanase produced by strain with deposit number MUCL 39199 (product referred as to L). The Panel could not conclude either on the efficacy of the products J, L as well as for the cellulase produced by the strain with deposit number CBS 120604 (product referred as to H) and the xylanase produced by the strain with deposit number CBS 614.94 (product referred to as K). The newly submitted information showed that the species for the xylanase produced by the strain with deposit number MUCL 39203 (product J), should be changed into Trichoderma citrinoviride, and that the species of the production strain for the glucanase produced by the strain with deposit number MUCL 39199 (product L), should be changed into Aspergillus neoniger. The Panel concluded that the four additives have the potential to reduce the protein degradation during the ensiling process when added at the corresponding inclusion levels in different types of forages.
Keywords: technological additives, silage, enzymes, efficacy
1. Introduction
1.1. Background and Terms of Reference as provided by the requestor
Regulation (EC) No 1831/2003 establishes the rules governing the Community authorisation of additives for use in animal nutrition and, in particular, Article 9 defines the terms of the authorisation by the Commission.
The applicant, FEFANA ASBL1, is seeking a Community authorisation of Enzymes as silage additives (7 types of enzymes) (SILAC) (12 strains) as a feed additive to be used as a silage additive for all animal species (Table 1).
Table 1.
Description of the substances
| Category of additive | Technological additives |
|---|---|
| Functional group of additive | Silage additives |
| Description | Enzymes as silage additives (7 types of enzymes) (SILAC) (12 strains) |
| Target animal category | All animal species |
| Applicant | FEFANA ASBL |
| Type of request | New opinion |
On 7 March 2018, the Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) of the European Food Safety Authority (“Authority”), in its opinion on the safety and efficacy of the product, could not conclude on alpha‐amylase from Bacillus amyloliquefaciens DSM 9553, Bacillus amyloliquefaciens NCIMB 30251, Aspergillus oryzae CBS 585.94 and Aspergillus oryzae ATTC SD‐5374, endo‐1,4‐beta‐glucanase from Trichoderma reesei ATCC PTA‐10001, Trichoderma reesei ATCC SD‐6331 and Aspergillus niger CBS 120604, endo‐1,4‐beta‐xyalanse from Trichoderma koningii MUCL 39203 and Trichoderma citrinoviride CBS 614.94 and endo‐1,3(4)‐beta‐glucanase from Aspergillus tubingensis MUCL 39199.
After the discussion with the Member States at the Standing Committee on Plants, Animals, Food and Feed, Section‐Animal Nutrition, from 27 to 29 June 2018, it was suggested to check for the possibility for the applicant to submit more data on:
Amylase from B. amyloliquefaciens NCIMB 30251 (in product C)
Cellulase from T. reesei ATCC PTA‐10001 (in product G)2
Cellulase from A. niger CBS 120604 (in product H)
Xylanase from T. koningii MUCL 39203 (product J)
Xylanase from T. longibrachiatum CBS 614.94 (product K) when used alone3
Glucanase from A. tubingensis MUCL 39199 (product L)
On amylase from A. oryzae CBS 585.94, EFSA could not conclude on the identity of the microorganism, therefore the conclusions on safety are limited.
The Commission gave the possibility to the applicant to submit complementary information and data in order to complete the assessment and to allow a revision of the Authority's opinion. The new data about alpha‐amylase produced by Aspergillus oryzae CBS 585.94; cellulase from Aspergillus niger CBS 120604 294; beta‐glucanase produced by Aspergillus niger MUCL 391994; xylanase produced by Trichoderma longibrachiatum MUCL 392035 and Xylanase produced by Trichoderma longibrachiatum CBS 614.946 have been received on 4 November 2020.
In view of the above, the Commission asks the Authority to deliver a new opinion on alpha‐amylase produced by Aspergillus oryzae CBS 585.94; cellulase from Aspergillus niger CBS 120604 294; beta‐glucanase produced by Aspergillus niger MUCL 391994; xylanase produced by Trichoderma longibrachiatum MUCL 392035 and Xylanase produced by Trichoderma longibrachiatum CBS 614.946 as feed additives for all animal species based on the additional data submitted by the applicant, in accordance with Article 29(1)(a) of Regulation 178/2002.
1.2. Additional information
In a previous opinion, the FEEDAP Panel evaluated 11 enzymes to be used as silage additives (EFSA FEEDAP Panel, 2018a). The applicant has now submitted supplementary information to complement the previous opinion. However, during the course of the current assessment the applicant withdrew the application for the alpha‐amylase produced by Aspergillus oryzae CBS 585.94.7
2. Data and methodologies
2.1. Data
The present assessment is based on data submitted by the applicant in the form of a supplementary information dossier8 to a previous application on the same additives9.
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.
2.2. Methodologies
The approach followed by the FEEDAP Panel to assess the safety and the efficacy of cellulase (produced by Aspergillus niger CBS 120604), beta‐glucanase (produced by Aspergillus neoniger MUCL 39199) and xylanases (produced by Trichoderma citrinoviride MUCL 39203 and CBS 614.94) is in line with the principles laid down in Regulation (EC) No 429/200810 and the relevant guidance documents: Guidance on the characterisation of microorganisms used as feed additives or as production organisms (EFSA FEEDAP Panel, 2018b) and the Guidance on the assessment of the efficacy of feed additives (EFSA FEEDAP Panel, 2018c).
3. Assessment
In a previous opinion, the FEEDAP Panel evaluated 11 enzymes to be used as silage additives (EFSA FEEDAP Panel, 2018a). In that opinion, the additives, including the production strains, were characterised in full and their safety and efficacy were evaluated. However, the Panel could not conclude on the identification of the production strains of the xylanase produced by the strain with deposit MUCL 39203 (referred as to product J) and the glucanase produced by the strain MUCL 39199 (referred as to product L). The Panel could neither conclude on the efficacy of the products J, L, nor for the cellulase produced by Aspergillus niger CBS 120604 (referred as to product H) and the xylanase produced by Trichoderma citrinoviride CBS 614.94 (referred to as product K).
The applicant has submitted new data to support the identification of the production strains of products J and L and the efficacy of the products J, L, H and K.
3.1. Characterisation of the production strains
The applicant submitted further information regarding the taxonomic identification of the production strains of products J and L.
The xylanase in product J is produced by a non‐genetically modified strain of Trichoderma deposited at the Culture collection Mycothèque de l'Université Catholique de Louvain (BCCM/MUCL) with the number MUCL 39203.11 The strain was originally identified as Trichoderma longibrachiatum MUCL 39203. In the assessment finalised in 2018, the strain was notified as Trichoderma koningii, but the Panel could not conclude on the identification due to the lack of sufficient information. In the current assessment, the production strain was identified by phylogenetic analysis of the translation elongation factor 1‐alpha (EF1‐α) DNA sequence and was shown to belong to Trichoderma citrinoviride species.12
The glucanase in product L is produced by a non‐genetically modified strain of Aspergillus deposited at the BCCM/MUCL with the number MUCL 39199.13 The strain was originally identified as Aspergillus niger. In the assessment finalised in 2018, the strain was notified as Aspergillus tubingensis, but the Panel could not conclude on the identification due to the lack of sufficient information. In the current assessment, the production strain was identified by phylogenetic analysis based on the calmodulin (CaM) gene as Aspergillus neoniger.14
The applicant provided also an updated certificate of deposition for the strain producing the product H, Aspergillus niger CBS 120604.15 For the strain producing the product K, Trichoderma citrinoviride (CBS 614.94), a certificate on the taxonomic classification of the strain and an updated certificate of deposition were provided.16
In the previous opinion, the FEEDAP Panel concluded that the use of these products in the production of silage are safe for the target animals, consumers and the environment. The new data on the taxonomic identification would not modify those conclusions.
3.2. Efficacy
The additives are to be used in all categories of forages for all animal species at an intended level per kg forage of 3.0 DNS17 U product H, 15 DNS U product K, 3.2 DNS U product J and 3.4 DNS U product L. A total of nine laboratory efficacy studies were submitted for products H and K,18 and four studies for products J and L. However, one of the studies for products H and K19 analysed the effect of the enzymes when added to forage together with lactic acid bacteria compared to a non‐supplemented control and was not considered further.
For products H and K, eight studies were conducted with different samples representing materials in the categories easy to ensile (studies 120, 221 and 322), moderately difficult to ensile (studies 423, 524 and 6 22 ) and difficult to ensile (studies 725 and 8 22 ), whereas for products J and L, three studies were conducted in moderately difficult (studies 926, 1027 and 1128) and one in difficult (study 12)29 to ensile forages as specified by Regulation (EC) No 429/2008 (Table 2).
Table 2.
Characteristics of the forage samples used in the 12 studies submitted
| Study | Test products | Test material (category) | Dry matter content (%) | Water‐soluble carbohydrate content (% fresh matter) |
|---|---|---|---|---|
| 1 | H, K | Grass (easy) | 40.3 | 4.1 |
| 2 | H, K | Grass (easy) | 34.9 | 4.1 |
| 3 | H, K | Lucerne (easy) | 39.2 | 3.2 |
| 4 | H, K | Maize (moderate) | 30.6 | 1.8 |
| 5 | H, K | Maize (moderate) | 30.5 | 2.4 |
| 6 | H, K | Lucerne (moderate) | 24.8 | 2.1 |
| 7 | H, K | Lucerne (difficult) | 18.9 | 1.2 |
| 8 | H, K | Lucerne (difficult) | 28.6 | 1.3 |
| 9 | J, L | Grass (moderate) | 37.0 | 2.2 |
| 10 | J, L | Lucerne (moderate) | 38.5 | 2.4 |
| 11 | J, L | Lucerne (moderate) | 21.0 | 1.5 |
| 12 | J, L | Alfalfa (difficult) | 23.7 | 1.1 |
All the studies included three treatment groups, a control and each of the enzymes tested at the corresponding recommended levels. The level applied to the forage was 3, 15, 3.2 and 3.4 DNS U/kg fresh forage for the products H, K, J and L, respectively, in accordance with the conditions of use. An aqueous suspension of the test item was prepared and then sprayed on the forage prior to ensiling. In the control silos, the same volume of water was added but without the additive. It is noted that in study 12 an authorised silage additive (1 k2106) was added to the forages in all groups (control and treated with J or L products). In all studies, forage was ensiled in mini‐silos with a capacity for more than 1 kg forage. The experiments were conducted in a cool dry place (products H and K, temperature not indicated) or at controlled temperature of 20–22°C (products J and L). The number of replicates per group (control and treated groups) in the studies for products H and K was 15, and in the studies for products J and L was 10 replicates in study 9, 5 in studies 10 and 11 and 4 in study 12.
Silos were opened after 90 days in all studies. The contents were analysed for dry matter (DM) content of the silage, pH, lactic acid, acetic acid, butyric acid (only for studies 3, 6 and 8) and ammonia. In studies 1–9, the final DM content of silage were corrected for volatiles accounting for the content of volatile fatty acids (including lactic acid) and alcohols with different formulas depending on the substrate used. For studies 10–12, the correction was done using a fixed correction for substrates depending on the one used in the study.30 An ANOVA followed by Tukey’s test was used for multiple comparison for studies with the products H and K, and Mann–Whitney was used for studies with products J and L to compare treated vs control silos. In all cases, significance was declared at p < 0.05. Results are shown in Tables 3 and 4.
Table 3.
Effects of products H and K on the characteristics of the silage at the end of the ensiling period (90 days)
| Study | Product (DNS U/kg forage) | DM content (%) | pH | Lactic acid (% DM) | Acetic acid (% DM) | Ammonia‐N as % of nitrogen |
|---|---|---|---|---|---|---|
| 1 (easy) | Control | 35.3 | 3.80 | 7.42 | 2.18 | 5.73 |
| H (3 U/kg) | 37.7* | 3.80 | 7.46 | 2.18 | 5.79 | |
| K (15 U/kg) | 38.3* | 3.83 | 7.26 | 2.45* | 6.62 | |
| 2 (easy) | Control | 33.0 | 3.96 | 6.43 | 2.17 | 1.05 |
| H (3 U/kg) | 35.8* | 4.04* | 6.06* | 2.09 | 0.70* | |
| K (15 U/kg) | 36.2* | 4.04* | 6.24* | 2.06* | 1.14 | |
| 3 (easy) | Control | 34.6 | 4.83 | 5.46 | 4.19 | 11.27 |
| H (3 U/kg) | 35.7* | 4.85 | 5.05 | 4.27 | 10.19* | |
| K (15 U/kg) | 36.4* | 4.84 | 5.26 | 3.95* | 10.17* | |
| 4 (moderate) | Control | 29.8 | 3.78 | 4.82 | 2.03 | 7.57 |
| H (3 U/kg) | 31.2* | 3.73* | 5.16* | 2.34* | 8.65 | |
| K (15 U/kg) | 30.8* | 3.73* | 5.19* | 2.48* | 8.44 | |
| 5 (moderate) | Control | 29.6 | 3.63 | 6.37 | 2.61 | 7.63 |
| H (3 U/kg) | 29.3 | 3.62 | 6.63* | 2.47 | 7.40* | |
| K (15 U/kg) | 28.9* | 3.64 | 6.53 | 2.41* | 6.96* | |
| 6 (moderate) | Control | 21.9 | 4.57 | 7.02 | 4.32 | 13.29 |
| H (3 U/kg) | 21.5 | 4.50 | 7.26 | 4.05 | 12.15* | |
| K (15 U/kg) | 22.3* | 4.56 | 6.89 | 4.27 | 12.11* | |
| 7 (difficult) | Control | 19.5 | 5.37 | 3.55 | 7.99 | 16.8 |
| H (3 U/kg) | 18.9 | 5.49 | 2.96 | 7.76 | 18.8 | |
| K (15 U/kg) | 18.2* | 5.58* | 3.00 | 8.03 | 19.5 | |
| 8 (difficult) | Control | 25.3 | 4.76 | 6.54 | 5.22 | 14.5 |
| H (3 U/kg) | 26.3* | 4.88 | 5.59 | 5.43 | 12.8* | |
| K (15 U/kg) | 30.2* | 4.82 | 5.74 | 4.63* | 6.4* |
DM: dry matter.
Means in a column within a given study are significantly different to the control p < 0.05.
Table 4.
Effects of products J and L on the characteristics of the silage at the end of the ensiling period (90 days)
| Study | Product (DNS U/kg forage) | DM content (%) | pH | Lactic acid (% DM) | Acetic acid (% DM) | Ammonia‐N as % of nitrogen |
|---|---|---|---|---|---|---|
| 9 (moderate) | Control | 39.9 | 4.28 | 4.97 | 1.40 | 7.7 |
| J (3.2 U/kg) | 38.7* | 4.26 | 4.89 | 1.33 | 7.4 | |
| L (3.4 U/kg) | 37.1* | 4.32 | 5.06 | 1.24 | 7.0* | |
| 10 (moderate) | Control | 37.6 | 4.15 | 4.80 | 2.65 | 18.9 |
| J (3.2 U/kg) | 38.0 | 3.99 | 5.34 | 2.59 | 17.3* | |
| L (3.4 U/kg) | 38.0 | 3.96 | 5.25 | 2.55 | 16.4* | |
| 11 (moderate) | Control | 19.5 | 3.62 | 5.74 | 3.16 | 24.4 |
| J (3.2 U/kg) | 19.9 | 3.70 | 6.84* | 3.23 | 22.1* | |
| L (3.4 U/kg) | 20.3 | 3.53 | 6.36* | 3.05 | 20.9* | |
| 12 (difficult) | Control + LAB | 20.5 | 4.69 | 10.2 | 5.76 | 18.5 |
| J (3.2 U/kg) + LAB | 19.9* | 4.68* | 16.6* | 6.17 | 15.7* | |
| L (3.4 U/kg) + LAB | 20.1* | 4.74* | 16.0* | 5.86 | 15.2* |
DM: dry matter.
Means in a column within a given study are significantly different to the control p < 0.05.
The results on the DM content at the end of the ensiling process and DM loss during ensiling (corrected for volatiles) were submitted by the applicant. However, the results showed inconsistencies and some of the values were unrealistic, which were probably due to the estimates used in the correction. Due to the high level of uncertainty, the Panel could not consider these data, and any significant differences observed in DM content or calculated DM loss were not considered in the assessment.
In forages treated with products H or K, the content of ammonia‐N (expressed as percent of total nitrogen) was significantly lower compared to the control in 5 out of 8 studies for product H and in 4 out of 8 for product K covering forages easy, moderately difficult and difficult to ensile. These results would indicate that the enzymes could contribute to reduce protein degradation of forage during the ensiling process. Other positive effects were also identified but showed no consistent pattern between studies.
In forages treated only with products J or L, the content of ammonia‐N (expressed as percent of total nitrogen) was significantly lower compared to the control in all four studies for product L and in three studies for product J. These results would indicate that the enzymes could contribute to reduce protein degradation of forage during the ensiling process. The lower ammonia‐N was observed in moderate to difficult to ensile forages with a DM content in the range between 21% and 40% before ensiling.
3.2.1. Conclusions on efficacy
Based on the results from 8 studies, the FEEDAP Panel concludes that the cellulase produced by Aspergillus niger CBS 120604 (product H) and the xylanase produced by Trichoderma citrinoviride CBS 614.94 (product K) at the proposed inclusion level (3 or 15 U/kg forage, respectively), have the potential to decrease protein degradation during ensiling in easy, moderately difficult and difficult to ensile forages.
Based on the results from four studies, the FEEDAP Panel concludes that the xylanase produced by Trichoderma citrinoviride MUCL 39203 (product J) and the glucanase produced by Aspergillus neoniger MUCL 39199 (product L) have the potential to decrease protein degradation during the ensiling process at 3.2 and 3.4 U/kg forage respectively. The effect has been identified in forages with DM content in the range between 21% and 40% before ensiling.
4. Conclusions
The species of the production strain (deposit number MUCL 39203) for the xylanase (product J) is Trichoderma citrinoviride. The species of the production strain (deposit number MUCL 39199) for the glucanase (product L) is Aspergillus neoniger.
The cellulase produced by Aspergillus niger CBS 120604 (product H), and the xylanase produced by Trichoderma citrinoviride CBS 614.94 (product K), at the proposed inclusion level (3 or 15 U/kg forage, respectively), have the potential to reduce the protein degradation during ensiling of easy, moderately difficult and difficult to ensile forages.
The xylanase produced by Trichoderma citrinoviride MUCL 39203 (product J) and the glucanase produced by Aspergillus neoniger MUCL 39199 (product L), at the proposed inclusion level (3.2 and 3.4 U/kg forage, respectively), have the potential to reduce the protein degradation during ensiling in forages with a DM content in the range between 21 and 40% before ensiling.
5. Documentation provided to EFSA/Chronology
| Date | Event |
|---|---|
| 03/11/2020 | Dossier received by EFSA. Dossier on enzymes as silage additives (7 types of enzymes) FEFANA ABSL. Submitted by SILAC, FEFANA ABSL |
| 25/11/2020 | Reception mandate from the European Commission |
| 17/05/2021 | 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 and efficacy |
| 17/09/2021 | Reception of supplementary information from the applicant ‐ Scientific assessment re‐started |
| 18/11/2021 | Request of supplementary information to the applicant in line with Article 8(1)(2) of Regulation (EC) No 1831/2003 – Scientific assessment suspended. Issues: efficacy |
| 13/01/2022 | Reception of supplementary information from the applicant ‐ Scientific assessment re‐started |
| 27/04/2022 | 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 and efficacy |
| 06/05/2022 | Reception of supplementary information from the applicant – Scientific assessment re‐started |
| 29/06/2022 | Opinion adopted by the FEEDAP Panel. End of the Scientific assessment |
Abbreviations
- BCCM/MUCL
Culture collection Mycothèque de l’Université Catholique de Louvain
- CaM
calmodulin
- DM
dry matter
- FEEDAP
EFSA Panel on Additives and Products or Substances used in Animal Feed
Suggested Citation: EFSA FEEDAP Panel (EFSA Panel on Additives and Products or Substances used in Animal Feed) , Bampidis V, Azimonti G, Bastos ML, Christensen H, Dusemund B, Fašmon Durjava M, Kouba M, López‐Alonso M, López Puente S, Marcon F, Mayo B, Pechová A, Petkova M, Ramos F, Sanz Y, Villa RE, Woutersen R, Dierick N, Galobart J, Revez J and Anguita M, 2022. Scientific Opinion on the safety and efficacy of the feed additives containing cellulase (produced by Aspergillus niger CBS 120604), beta‐glucanase (produced by Aspergillus neoniger MUCL 39199), or xylanase (produced by Trichoderma citrinoviride MUCL 39203 or by Trichoderma citrinoviride CBS 614.94) for all animal species (FEFANA ASBL). EFSA Journal 2022;20(8):7425, 10 pp. 10.2903/j.efsa.2022.7425
Requestor: European Commission
Question numbers: EFSA‐Q‐2020‐00836
Panel members: Vasileios Bampidis, Giovanna Azimonti, Maria de Lourdes Bastos, Henrik Christensen, Birgit Dusemund, Mojca Fašmon Durjava, Maryline Kouba, Marta López‐Alonso, Secundino López Puente, Francesca Marcon, Baltasar Mayo, Alena Pechová, Mariana Petkova, Fernando Ramos, Yolanda Sanz, Roberto Edoardo Villa and Ruud Woutersen.
Declarations of interest: If you wish to access the declaration of interests of any expert contributing to an EFSA scientific assessment, please contact interestmanagement@efsa.europa.eu.
Adopted: 29 June 2022
Notes
Rue de Trèves 45, 1,040 Brussels (Belgium).
It is noted that the cellulase in product G was from T. reesei ATCC SD 6331.
Identified in FEEDAP opinion as Trichoderma citrinoviride.
Indentified in FEEDAP Opinion as Aspergillus tubingensis.
Indentified in FEEDAP Opinion as Trichoderma koningii.
Indentified in FEEDAP Opinion as Trichoderma citrinoviride.
The application for amylase from A. oryzae CBS 585.94 was withdrawn by the applicant and officially communicated to EFSA on 3 December 2021.
FEED dossier reference: FAD‐2020‐0088.
FEED dossier reference: FAD‐2010‐0367.
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.
Technical dossier FAD‐2020‐0088/Supplementary information May 2022/Annex 1.
Technical dossier FAD‐2020‐0088/Annex 3a and supplementary information September 2021.
Technical dossier FAD‐2020‐0088/Annex 3c and supplementary information September 2021/Annex 1.
Technical dossier FAD‐2020‐0088/Annex 3b and c and Supplementary information September 2021 Annex 1.
Technical dossier FAD‐2020‐0088/Annex 4a.
Technical dossier FAD‐2020‐0088/Annex 4b and Supplementary information May 2022/Annex 2.
DNS U is the amount of reducing sugar released by a glucanase/xylanase as glucose/xylose equivalents in μmol/g per min at pH 4.5 and 37°C from carboxymethil cellulose/birchwood xylan.
Technical dossier FAD‐2020‐0088/Annex 5a and Supplementary information September 2021 and January 2022.
Technical dossier FAD‐2020‐0088/ Annex 5h.
Technical dossier FAD/2020‐0088/Annex 5b Trial Report 1and Supplementary information September 2021 and January 2022.
Technical dossier FAD‐2020‐0088/Annex 5d Trial Report 3 and Supplementary information September 2021 and January 2022.
Technical dossier FAD‐2020‐0088/Annex 5g Trial Report 6 and Supplementary information September 2021 and January 2022.
Technical dossier FAD‐2020‐0088/Annex 5c Trial Report 2 and Supplementary information September 2021 and January 2022.
Technical dossier FAD‐2020‐0088/Annex 5f Trial Report 5 and Supplementary information September 2021 and January 2022.
Technical dossier FAD‐2020‐0088/Annex 5e Trial Report 4 and Supplementary information September 2021 and January 2022.
Technical dossier FAD‐2020‐0088/Main text and Annex 6d and 6e and 7c and Supplementary information September 2021/annex 3 and 8 and Supplementary information January 2022 and May 2022.
Technical dossier FAD‐2020‐0088/Main text and Annex 6d and 6e and 7d study 3 and Supplementary information September 2021/annex 4 and 9 and Supplementary information January 2022 and May 2022.
Technical dossier FAD‐2020‐0088/Main text and Annex 6d and 6e and 7e and Supplementary information September 2021/annex 5 and 10 and Supplementary information January 2022 and May 2022.
Technical dossier FAD‐2020‐0088/Main text and Annexes 6a to 6c, 7b Supplementary information September 2021/annex 2 and 7 and Supplementary information January 2022 and May 2022.
Technical dossier FAD‐2020‐0088/Supplementary information September 2021/Annex 6.
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