Marta López Alonso

Safety and efficacy of Bacillus subtilis PB6 (Bacillus subtilis ATCC PTA‐6737) as a feed additive for sows

Abstract Bacillus subtilis PB6 is a feed additive based on viable spores of a strain of Bacillus subtilis. EFSA has already issued several opinions on the safety and efficacy of the additive when used in the feed for a number of avian and porcine species/categories. The applicant is now seeking authorisation for use in sows in order to have benefits in piglets. In the course of previous assessments, no evidence of a toxigenic potential or resistance to relevant antibiotics was found. Consequently, the strain of B. subtilis in the additive, following the qualified presumption of safety approach to safety assessment, is presumed safe for target animals including sows and their offspring, consumers and the environment. In a previous assessment, it was also concluded that the additive is not a skin/eye irritant or a skin sensitiser and that there were no concerns on respiratory sensitisation. The use of the additive with feed for sows is considered unlikely to introduce hazards for users of the product not already considered. Five studies are described in which groups of sows given the additive for a minimum period from the last 3 weeks of pregnancy, throughout farrowing and lactation, until weaning of piglets were compared with a control group. In only two of five studies, there was a significant beneficial effect seen in terms of piglet performance. However, when data from the four similar were pooled and analysed, significant increases in weaned weight and average daily gain were indicated for piglets from sows given the additive. Consequently, the FEEDAP Panel concludes that Bacillus subtilis PB6 when added to diets of sows from 3 weeks before parturition until weaning of piglets at a dose of 1 × 108 CFU/kg complete feed has the potential to improve the growth of piglets from birth to weaning.

Safety and efficacy of Coxar® (nicarbazin) for turkeys for fattening

Abstract The coccidiostat Coxar® is safe for turkeys for fattening at the use level of 100 mg nicarbazin/kg complete feed, with a margin of safety of about 1.25. Nicarbazin, when ingested, is rapidly split in its two components 2‐hydroxy‐4,6‐dimethylpyrimidine (HDP) and dinitrocarbanilide (DNC), which behave independently. HDP‐related residues are much lower than those of DNC. DNC is the marker residue. Liver is the target tissue. Nicarbazin is not genotoxic. The primary toxicity resulting from the oral use of nicarbazin is renal toxicity. The lowest no observed adverse effect level (NOAEL) identified in a 52‐week study in rat using DNC+HDP is 20 mg DNC + 8 mg HDP/kg body weight (bw) per day based on the absence of microcrystals in urine and related microscopic renal observations. The use of 100 mg nicarbazin from Coxar®/kg complete feed for turkeys for fattening will not pose a risk to consumers, provided that maximum contents in nicarbazin of 0.1% p‐nitroaniline (PNA) and 0.4% methyl(4‐nitrophenyl) carbamate (M4NPC) would be respected. No withdrawal time is required. Residue data comply with the established maximum residue limits (MRLs). Nicarbazin is not a skin or eye irritant and not a skin sensitiser. These conclusions also apply to the additive Coxar®. Inhalation toxicity of nicarbazin is limited; the granulated additive has a low dusting potential. No risk for users is identified. Based on the available data, the FEEDAP Panel cannot conclude on the safety of Coxar® for the environment. The efficacy of 100 mg nicarbazin from Coxar®/kg feed was demonstrated in three anticoccidial sensitivity tests (AST), but only in one floor pen study. The floor pen study with 75 mg nicarbazin failed to demonstrate evidence of efficacy. No final conclusions on the efficacy of nicarbazin from Coxar® for turkeys for fattening can be drawn.

Safety and efficacy of Coxiril® (diclazuril) for chickens reared for laying

Abstract 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 Coxiril® (diclazuril) for chickens reared for laying. Coxiril®, containing 0.5% diclazuril, is intended for the prevention of coccidiosis in chickens reared for laying at a dose range of 0.8–1.2 mg diclazuril/kg of complete feed up to a maximum age of 12 weeks. Derived from data already assessed for chickens and turkeys for fattening, diclazuril from Coxiril® is safe for chickens reared for laying up to 1.2 mg/kg complete feed when applied until 12 weeks of age. The FEEDAP Panel extended its previous assessment of consumer safety for the use of diclazuril from Coxiril® in chickens for fattening to chickens reared for laying. No measurable diclazuril residues were found in the first eggs laid from chickens reared for laying fed diclazuril from Coxiril® at 1.2 mg/kg complete feed until 12 weeks of age. Coxiril® was considered as a non‐irritant to eyes and skin. It is not a potential skin sensitiser. User inhalation exposure to Coxiril®, as a result of normal handling, is unlikely to cause respiratory or systemic toxicity. The use of diclazuril from Coxiril® in chickens reared for laying at the highest proposed feed concentration would not pose a risk to the environment for neutral/alkaline soils (pH ≥ 7). A final conclusion on the risk resulting from the use of diclazuril in acid soil from Coxiril® cannot be done due to the high uncertainties related to potential accumulation of diclazuril over time. Derived from data already assessed for chickens for fattening, diclazuril from Coxiril® has the potential to control coccidiosis in chickens reared for laying at a minimum concentration of 0.8 mg/kg complete feed.

Efficacy of Saccharomyces cerevisiae (NBRC 0203), Lactobacillus plantarum (NBRC 3070) and Lactobacillus casei (NBRC 3425) as a silage additive for all species

Abstract The product under assessment is a preparation containing single strains of Saccharomyces cerevisiae, Lactobacillus plantarum and Lactobacillus casei to be used as a technological additive to improve the ensiling process. EFSA has been previously requested by the European Commission to evaluate this product. The safety of the additive for consumers, users, the environment and target animals was established at that time. Results from a number of in vitro ensiling studies made using a variety of grass forage and maize samples were also presented. These efficacy studies showed no evidence that the additive had the potential to conserve nutrients and so improve the production of silage. There was, however, evidence for improved aerobic stability once the ensiled material was exposed to air, but because of the apparent variability in the additive mix, an effective dose could not be established. The applicant has now provided a number of additional ensiling studies confirming that the additive when applied at 80 mL/tonne significantly increases the aerobic stability of ensiled material after exposure to air. This was shown in forage materials with dry matter contents varying between 30% and 70%. Fluorescence in situ hybridisation methods were also introduced to better characterise the additive. Although this method allows individual strains to be separately enumerated, bacterial and yeast numbers are expressed as cell numbers/mL and include counts of both viable and non‐viable cells. As a result values cannot be equated to colony‐forming units, the basis of any authorisation and the functional properties of the additive. Consequently, the Panel remains unable to identify a minimum specification for the product or a minimum effective dose.

Safety and efficacy of zinc chelate of methionine sulfate for all animal species

Abstract The additive ‘Zinc chelate of methionine sulfate’ is zinc chelated with methionine in a molar ratio 1:1, with a minimum zinc content of 19.1%. Owing to the limitations of the tolerance study, it could not be used for the assessment of safety for target animals; therefore, the EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP Panel) cannot conclude on the safety of zinc chelate of methionine sulfate for the target species. No concerns for consumer safety are expected from the use of the zinc chelate of methionine sulfate in animal nutrition when used up to the maximum EU authorised zinc levels in feed. Zinc chelate of methionine sulfate should be considered as a skin and eye irritant, and as a skin sensitiser; it is considered to pose a risk by inhalation to the users. The additive under assessment, zinc chelate of methionine sulfate, is intended to be a substitute for other authorised zinc additives and will not further increase the environmental burden of zinc; therefore, the FEEDAP Panel considers that the use of the additive in animal nutrition would not pose an additional risk for the environment. Based on literature studies and a specific study conducted with the additive under assessment, zinc chelate of methionine sulfate is an available source of zinc for all animal species.

Safety and efficacy of Bacillus amyloliquefaciens (NCIMB 30229) as a silage additive for all animal species

Abstract The additive under assessment is a preparation of Bacillus amyloliquefaciens NCIMB 3022 intended for use as a technological additive in forages for all animal species at a proposed minimum dose of 5 × 107 CFU/kg fresh materials. The species B. amyloliquefaciens is considered by EFSA to be suitable for the qualified presumption of safety approach to safety assessment. This approach requires the identification of the strain and evidence that it is not toxigenic and does not show acquired resistance to relevant antibiotics. In a previous assessment, the identity and susceptibility to clinically relevant antibiotics of the active agent was established but the lack of toxigenic potential could not be demonstrated. Therefore, the Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) could not conclude on the safety of the product for target animals, consumers, users or the environment. In the same opinion, the potential of the additive to improve aerobic stability of silages at the proposed dose was not convincingly demonstrated. A new cytotoxicity study was conducted using methanol extracts. The methanol extract was cytotoxic at the inclusion level of 5%, while no cytotoxicity was observed at lower concentrations. The current guidance on Bacillus requires absence of toxic effect using the non‐concentrated supernatant. Since the applicant was unable to determine the correlation between the methanol extracts and the Bacillus supernatant concentrations, the FEEDAP Panel is unable to conclude on the toxigenic potential of the strain based on the current data. An additional efficacy study with a similar protocol to the ones previously assessed was conducted. The study showed an improved aerobic stability when added to forage at the proposed dose. However, the FEEDAP Panel remains unable to conclude on the efficacy of the additive to improve the ensiling process on the basis of a single positive result.

Safety and efficacy of OPTIPHOS® (6-phytase) as a feed additive for finfish

Abstract The additive OPTIPHOS ® is a preparation of 6‐phytase that is authorised for use in avian species, weaned piglets, pigs for fattening and sows. The applicant seeks for an extension of use of the product to finfish, at a dose range between 250 and 1,000 OTU/kg feed. The FEEDAP Panel concluded in a previous opinion that there are no concerns for consumer safety and no risks for the environment are expected from the use of the product as feed additive. Moreover, it was concluded that the additive is not a skin/eye irritant or a skin sensitiser, but has the potential to be a respiratory sensitiser. The Panel considered that the new use of the additive would not change the previous conclusions regarding the safety for the consumer, user and environment. The results of a tolerance study performed in juvenile rainbow trout (Oncorhynchus mykiss) showed no negative effects of the additive on the fish when offered up to 100 times the maximum recommended dose. Therefore, the Panel concluded that the additive is safe for the rainbow trout and extrapolated this conclusion to all finfish. The Panel evaluated three efficacy trials, two performed in rainbow trout and one in Atlantic salmon (Salmo salar). In these studies, the performance of the fish was monitored as well as phosphorus digestibility and phosphorus retention parameters. In the three trials, the performance and the phosphorus retention were improved by the additive at the lowest tested dose (in rainbow trouts at 250 OTU/kg feed and in Atlantic salmon at 500 OTU/kg feed). The FEEDAP Panel concluded that the additive has a potential to be efficacious in rainbow trout and salmon at 500 OTU/kg. The Panel extrapolated the conclusion to all finfish species.

Safety and efficacy of Levucell® SB (Saccharomyces cerevisiae CNCM I‐1079) as a feed additive for chickens for fattening and minor poultry species

Abstract Levucell SB ® is a feed additive consisting of viable cells of a strain of Saccharomyces cerevisiae currently authorised as a zootechnical additive for piglets and sows. The applicant is now seeking authorisation as a zootechnical additive (other zootechnical additive) for use with chickens for fattening and minor poultry species. The EFSA FEEDAP Panel in a previous opinion concluded that the additive fulfilled the requirements for the qualified presumption of safety (QPS) approach to safety assessment and thus could be presumed safe for the target species, consumers and the environment. Since the additive which is the subject of the present application has the same formulations as that considered previously, following the QPS approach, the same conclusions on target animal consumer and environmental safety apply when used with poultry. The use of the additive with diets for the new target species is considered unlikely to introduce hazards for users of the product not already considered in the previous assessment. When used with poultry feed, the additive is intended to aid the reduction of carcass contamination with enteropathogens and so improve the quality of poultry products. Five studies were submitted with chickens fed diets with or without the additive and then (in four studies) artificially challenged with Salmonella. The fifth study relied on the natural incidence of Salmonella. The prevalence of Salmonella contamination was estimated by examining the carcass directly and/or by detection in caecal, cloacal or faecal samples. Considering overall the results of the five studies, there is evidence that the addition of Levucell SB ® to diets at a dose of 2 × 1010 CFU/kg feed can aid the reduction of carcass contamination with Salmonella spp.. This conclusion can be extrapolated to minor avian species for fattening when used at the same dose, but not to minor poultry species for laying.

Safety and efficacy of Hemicell® HT (endo‐1,4‐β‐d‐mannanase) as a feed additive for chickens for fattening, chickens reared for laying, turkey for fattening, turkeys reared for breeding, weaned piglets, pigs for fattening and minor poultry and porcine species

Abstract Hemicell® HT is a feed additive with endo‐1,4‐β‐d‐mannanase as the main enzymatic activity that is available in solid (HT) and liquid (HT‐L) forms. The production strain of the enzyme is a genetically modified strain of Paenibacillus lentus. The recipient strain is considered to be safe, the sequences introduced to obtain the production strain do not raise safety concerns. The absence of the production strain and of recombinant DNA in the product was not proven. The additive is safe for the target species at the corresponding recommended doses. The use of Hemicell® HT as a feed additive does not give rise to concerns for consumers. Hemicell® HT and Hemicell® HT‐L are not irritant to the skin and eyes; Hemicell® HT is a skin sensitiser. No specific data were provided on the effects on the respiratory system; however, considering the nature of the active substance, the additive is considered a potential respiratory sensitiser. The active substance of the additive is a protein, and as such, it will be degraded/inactivated during passage through the digestive tract of the animals or in the environment. However, uncertainty remains on the absence of the production strain and of recombinant DNA in the product, consequently, the EFSA FEEDAP Panel could not conclude on the environmental safety of the product with regard to the genetically modified production strain. The additive has the potential to be efficacious in chickens for fattening at 32,000 U/kg feed and at 48,000 U/kg feed in turkeys for fattening and weaned piglets. These conclusions were extended to chickens reared for laying and turkeys reared for breeding and extrapolated to minor poultry species for fattening or reared for laying/breeding. The Panel could not conclude on the efficacy of the product in pigs for fattening or in minor porcine species.

Safety and efficacy of Lactobacillus acidophilus D2/CSL (Lactobacillus acidophilus CECT 4529) as a feed additive for chickens for fattening

Abstract 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 Lactobacillus acidophilus D2/CSL when used in feed for chickens for fattening at a minimum dose of 1 × 109 colony‐forming units (CFU)/kg complete feedingstuffs. The additive is a preparation of viable cells of L. acidophilus. This species is considered by EFSA to be suitable for the qualified presumption of safety (QPS) approach to establish safety for the target species, consumers and the environment. The safety of Lactobacillus acidophilus CECT 4529 was assessed by EFSA in 2014. Following the QPS approach to safety assessment, Lactobacillus acidophilus CECT 4529 is assumed to be safe for the target species, the consumer and the environment without the need for further studies. No concerns are expected from other excipients present in the product, so Lactobacillus acidophilus D2/CSL is also considered safe for target animals, including chickens for fattening, consumers and the environment. The safety of the additive for the user was also considered in that opinion. The FEEDAP Panel is unaware of any new data that would lead it to revise its conclusions that the additive should be considered to be an eye/skin irritant and a skin/respiratory sensitiser. There is insufficient evidence to conclude on the efficacy of the additive when used in diets for chickens for fattening.