Risks associated with endotoxins in feed additives produced by fermentation

Acknowledgements We thank Jordi Tarrés Call, who recorded the discussions and offered advice about procedures, and Nicole Reisinger and Gerd Schatzmayr, who provided valuable information about endotoxins in animal feeds. The Rowett Institute of Nutrition and Health is funded by the Rural and Environment Science and Analytical Services Division (RESAS) of the Scottish Government.Peer reviewedPublisher PD

Guidance on protocol development for EFSA generic scientific assessments

EFSA Strategy 2027 outlines the need for fit‐for‐purpose protocols for EFSA generic scientific assessments to aid in delivering trustworthy scientific advice. This EFSA Scientific Committee guidance document helps address this need by providing a harmonised and flexible framework for developing protocols for EFSA generic assessments. The guidance replaces the ‘Draft framework for protocol development for EFSA's scientific assessments’ published in 2020. The two main steps in protocol development are described. The first is problem formulation, which illustrates the objectives of the assessment. Here a new approach to translating the mandated Terms of Reference into scientifically answerable assessment questions and sub‐questions is proposed: the ‘APRIO' paradigm (Agent, Pathway, Receptor, Intervention and Output). Owing to its cross‐cutting nature, this paradigm is considered adaptable and broadly applicable within and across the various EFSA domains and, if applied using the definitions given in this guidance, is expected to help harmonise the problem formulation process and outputs and foster consistency in protocol development. APRIO may also overcome the difficulty of implementing some existing frameworks across the multiple EFSA disciplines, e.g. the PICO/PECO approach (Population, Intervention/Exposure, Comparator, Outcome). Therefore, although not mandatory, APRIO is recommended. The second step in protocol development is the specification of the evidence needs and the methods that will be applied for answering the assessment questions and sub‐questions, including uncertainty analysis. Five possible approaches to answering individual (sub‐)questions are outlined: using evidence from scientific literature and study reports; using data from databases other than bibliographic; using expert judgement informally collected or elicited via semi‐formal or formal expert knowledge elicitation processes; using mathematical/statistical models; and – not covered in this guidance – generating empirical evidence ex novo. The guidance is complemented by a standalone ‘template’ for EFSA protocols that guides the users step by step through the process of planning an EFSA scientific assessment

Re-evaluation of the existing health-based guidance values for copper and exposure assessment from all sources

Copper is an essential micronutrient and also a regulated product used in organic and in conventional farming pest management. Both deficiency and excessive exposure to copper can have adverse health effects. In this Scientific Opinion, the EFSA 2021 harmonised approach for establishing health-based guidance values (HBGVs) for substances that are regulated products and also nutrients was used to resolve the divergent existing HBGVs for copper. The tightly regulated homeostasis prevents toxicity manifestation in the short term, but the development of chronic copper toxicity is dependent on copper homeostasis and its tissue retention. Evidence from Wilson disease suggests that hepatic retention is indicative of potential future and possibly sudden onset of copper toxicity under conditions of continuous intake. Hence, emphasis was placed on copper retention as an early marker of potential adverse effects. The relationships between (a) chronic copper exposure and its retention in the body, particularly the liver, and (b) hepatic copper concentrations and evidence of toxicity were examined. The Scientific Committee (SC) concludes that no retention of copper is expected to occur with intake of 5 mg/day and established an Acceptable Daily Intake (ADI) of 0.07 mg/kg bw. A refined dietary exposure assessment was performed, assessing contribution from dietary and non-dietary sources. Background copper levels are a significant source of copper. The contribution of copper from its use as plant protection product (PPP), food and feed additives or fertilisers is negligible. The use of copper in fertilisers or PPPs contributes to copper accumulation in soil. Infant formula and follow-on formula are important contributors to dietary exposure of copper in infants and toddlers. Contribution from non-oral sources is negligible. Dietary exposure to total copper does not exceed the HBGV in adolescents, adults, elderly and the very elderly. Neither hepatic copper retention nor adverse effects are expected to occur from the estimated copper exposure in children due to higher nutrient requirements related to growth

Guidance on the use of the benchmark dose approach in risk assessment

The Scientific Committee (SC) reconfirms that the benchmark dose (BMD) approach is a scientifically more advanced method compared to the no-observed-adverse-effect-level (NOAEL) approach for deriving a Reference Point (RP). The major change compared to the previous Guidance (EFSA SC, 2017) concerns the Section 2.5, in which a change from the frequentist to the Bayesian paradigm is recommended. In the former, uncertainty about the unknown parameters is measured by confidence and significance levels, interpreted and calibrated under hypothetical repetition, while probability distributions are attached to the unknown parameters in the Bayesian approach, and the notion of probability is extended to reflect uncertainty of knowledge. In addition, the Bayesian approach can mimic a learning process and reflects the accumulation of knowledge over time. Model averaging is again recommended as the preferred method for estimating the BMD and calculating its credible interval. The set of default models to be used for BMD analysis has been reviewed and amended so that there is now a single set of models for quantal and continuous data. The flow chart guiding the reader step-by-step when performing a BMD analysis has also been updated, and a chapter comparing the frequentist to the Bayesian paradigm inserted. Also, when using Bayesian BMD modelling, the lower bound (BMDL) is to be considered as potential RP, and the upper bound (BMDU) is needed for establishing the BMDU/BMDL ratio reflecting the uncertainty in the BMD estimate. This updated guidance does not call for a general re-evaluation of previous assessments where the NOAEL approach or the BMD approach as described in the 2009 or 2017 Guidance was used, in particular when the exposure is clearly lower (e.g. more than one order of magnitude) than the health-based guidance value. Finally, the SC firmly reiterates to reconsider test guidelines given the wide application of the BMD approach

Safety and efficacy of a feed additive consisting of Bacillus velezensis PTA‐6507, B. velezensis NRRL B‐50013 and B. velezensis NRRL B‐50104 (Enviva® PRO 202 GT) for turkeys for fattening (Danisco Animal Nutrition)

[EN]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 the additive consisting of Bacillus amyloliquefaciens PTA-6507, B. amyloliquefaciens NRRL B-50013 and B. amyloliquefaciens NRRL B-50104 (trade name: Enviva® PRO 202 GT) for turkeys for fattening. The product under assessment is based on viable spores of three strains originally identified as B. amyloliquefaciens which, in the course of the current assessment, were reclassified as Bacillus velezensis. The bacterial species B. velezensis is considered suitable for the qualified presumption of safety (QPS) approach to safety assessment. The identity of the active agents was established. The active agents do not harbour acquired antimicrobial resistance genes and lack toxigenic potential and the capacity to produce aminoglycosides. Following the QPS approach, the three bacterial strains are presumed safe for the target species, consumers and the environment. Since no concerns are expected from the other components of the additive, Enviva® PRO 202 GT is also considered safe for the target species, consumers and the environment. Enviva® PRO 202 GT is non-irritant to skin and eyes and is not a dermal sensitiser. Due to the proteinaceous nature of the active agents, the additive should be considered a respiratory sensitiser. In a previous opinion, it was concluded that Enviva® PRO 202 GT has a potential to be efficacious as a zootechnical additive in chickens for fattening at the recommended level of 7.5 × 107 CFU/kg complete feed. It is considered that conclusions on efficacy of Enviva® PRO 202 GT in chickens for fattening can be extrapolated to turkeys for fattening. Therefore, the FEEDAP Panel concludes that Enviva® PRO 202 GT has the potential to be efficacious in turkeys for fattening at 7.5 × 107 CFU/kg complete feed.S

Safety and efficacy of a preparation of algae interspaced bentonite as a feed additive for all animal species

Following a request from the European Commission, the EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP Panel) was asked to deliver a scientific opinion on the safety and efficacy of a preparation of algae interspaced bentonite when used as a flatoxin B1 binder for all animal species. The additive is composed of bentonite and algae belonging to Ulva spp. The additive is considered safe for weaned piglets, dairy cows and chickens for fattening at the maximum recommended dose of 125 mg/kg complete feed (a wide margin of safety is established in weaned piglets and dairy cows); this conclusion is extrapolated to all animal species. The additive is not genotoxic. As bentonite is essentially not absorbed from the gut lumen and algae from Ulva spp. are not expected to be of concern for human consumption, the FEEDAP Panel considers that the use of the additive in animal nutrition is safe for consumers. The additive is not an irritant to the skin or the eyes and it is considered to have low inhalation toxicity. However, the additive has a high dusting potential and contains a high proportion offine particles. A high level of inhalation exposure to an inert dust may be hazardous. In the absence of data, the Panel could not conclude on dermal sensitisation. As the components of the additive are of natural origin (soil and marine environment), it is not expected that the use of the additive in animal nutrition would adversely affect the environment. The FEEDAP Panel could not conclude on the efficacy of the additive for all animal species

Safety of the fermentation product of Aspergillus oryzae NRRL 458 (Amaferm®) as a feed additive for dairy cows (Biozyme Inc.)

[EN] Amaferm® is a fermentation product produced by Aspergillus oryzae NRRL 458, containing alpha-amylase and cellulase enzyme activities, authorised for use as a feed additive for dairy cows. In 2016, the applicant requested for the renewal of the authorisation and the EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) issued an opinion at that regard in 2020. In that opinion, the Panel could not confirm the previously drawn conclusions (EFSA, 2006) regarding the safety of the production strain, and consequently could not confirm the safety of the additive for the target species and consumers. In the current submission, the applicant provided supplementary information that allowed the Panel to conclude on the identity of the production strain, redefine the specifications of the additive, and finally to conclude on its safety. Therefore, the new data provided permit to conclude that Amaferm® complies with the conditions of the authorisation. However, the Panel noted that there is the need to change the specification and description of the cellulase and amylase units in the authorisation act. The data provided in the previous (EFSA FEEDAP Panel, 2020) and the current assessments support that Amaferm® remains safe under the approved conditions for target species, consumers and the environment. The additive is non-irritant to skin and eyes, or a dermal sensitiser but should be considered a potential respiratory sensitiser.SIThe Panel wishes to thank the following for the support provided to this scientific output (in alphabetical order of the last name): Working Group on Animal Nutrition and Working Group on Microbiology, Rosella Brozzi, Christina Cuomo and Jordi Tarres Call

Assessment of the efficacy of a feed additive consisting of Limosilactobacillus reuteri (formerly Lactobacillus reuteri) DSM 32203 for dogs (NBF LANES)

Following a request from the European Commission, EFSA was asked to deliver a scientific opinion on the efficacy of Limosilactobacillus reuteri (formerly Lactobacillus reuteri) DSM 32203 as a zootechnical additive for dogs. The additive is a preparation of viable cells of L. reuteri DSM 32203 and it has not been previously authorised as a feed additive in the European Union. The additive is intended for use in complete feed for dogs at a minimum use level of 6 x 10(9) colony forming units (CFU) per animal and day. In a previous opinion, the FEEDAP Panel could not conclude on the efficacy of L. reuteri DSM 32203 for dogs. The applicant has provided supplementary information to support the efficacy of the additive to dogs. Based on the data provided, the FEEDAP Panel concludes that L. reuteri DSM 32203 has the potential to improve faecal consistency by reducing the moisture content of stools from dogs receiving the additive at 1 x 10(10) CFU/kg feed. However, the Panel has some reservations on the effects in the moisture content of stools, which if maintained over time might cast doubts on the benefits on the long-term use of the additive since it could lead to constipation

Safety and efficacy of a feed additive consisting of lactic acid produced by Weizmannia coagulans (synonym Bacillus coagulans) DSM 32789 for all animal species except for fish (Jungbunzlauer SA)

©2022EuropeanFoodSafetyAuthority.EFSAJournalpublishedbyWiley-VCHGmbHonbehalfofEuropeanFoodSafetyAuthority.[EN] 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 lactic acid produced by a non-genetically modified strain of Weizmannia coagulans (synonym of Bacillus coagulans) (DSM 32789) for all animal species except for fish. The production strain qualifies for the QPS approach for safety assessment. Although uncertainty remains concerning the possible presence of viable cells and/or spores of the production strain in the final product, this does not raise safety concerns for the target species, humans and the environment. The lactic acid is safe at 50,000 mg/kg complete feed for functional ruminants and pigs and at 20,000 mg/kg feed for all the other animal species and categories except for pre-ruminants for which a safe level cannot be established. The corresponding safe levels in water for drinking would be 15,000 mg/L water for pigs and 8,000 mg/L for other non-ruminant species. Although no safe concentration of lactic acid in water for drinking for ruminants can be derived, the Panel considers that the use in water for drinking is safe in ruminants when the total daily intake of the additive does not exceed the daily amount that is considered safe when consumed via feed. The use of the additive under assessment in animal nutrition is considered safe for the consumers and for the environment. It is considered corrosive to the skin, eyes and mucous membranes. Lactic acid is used in food as a preservative. It is reasonable to expect that the effect seen in food will be observed in feed when it is used at comparable concentrations and conditions. However, the FEEDAP Panel has reservations about its effectiveness as a preservative in complete feed with a moisture content of ≤ 12%.SIThe Panel wishes to acknowledge the contribution to this opinion of Martina Reitano, Joana Revez, the experts of the Microbiology and of the Animal Nutrition Working Groups of the FEEDAP Panel

Safety and efficacy of a preparation of algae interspaced bentonite as a feed additive for all animal species

Following a request from the European Commission, the EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP Panel) was asked to deliver a scientific opinion on the safety and efficacy of a preparation of algae interspaced bentonite when used as aflatoxin B1 binder for all animal species. The additive is composed of bentonite and algae belonging to Ulva spp. The additive is considered safe for weaned piglets, dairy cows and chickens for fattening at the maximum recommended dose of 125 mg/kg complete feed (a wide margin of safety is established in weaned piglets and dairy cows); this conclusion is extrapolated to all animal species. The additive is not genotoxic. As bentonite is essentially not absorbed from the gut lumen and algae from Ulva spp. are not expected to be of concern for human consumption, the FEEDAP Panel considers that the use of the additive in animal nutrition is safe for consumers. The additive is not an irritant to the skin or the eyes and it is considered to have low inhalation toxicity. However, the additive has a high dusting potential and contains a high proportion of fine particles. A high level of inhalation exposure to an inert dust may be hazardous. In the absence of data, the Panel could not conclude on dermal sensitisation. As the components of the additive are of natural origin (soil and marine environment), it is not expected that the use of the additive in animal nutrition would adversely affect the environment. The FEEDAP Panel could not conclude on the efficacy of the additive for all animal species