SCIENTIFIC OPINION. EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) 2,3

Transcription

1 SCIENTIFIC OPINION Scientific Opinion on the safety and efficacy of Calsporin (Bacillus subtilis) for turkeys for fattening, ducks, geese, pigeons and other game birds for meat production, ducks, geese, pigeons, game birds, ornamental and sporting birds for rearing to point of lay, turkeys reared for breeding and chickens reared for laying 1 EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) 2,3 ABSTRACT European Food Safety Authority (EFSA), Parma, Italy Calsporin is a feed additive based on a strain of Bacillus subtilis. The applicant is seeking authorisation for the use of the product in feeds for turkeys for fattening, ducks, geese, pigeons and other game birds for meat production, ducks, geese, pigeons, game birds, ornamental and sporting birds for rearing to point of lay, and turkeys reared for breeding at the dose of 3 x 10 8 CFU/kg feed, and for chickens reared for laying at the dose of 5 x 10 8 CFU/kg feed. EFSA has been requested to deliver an opinion on the safety of the product for target animals, consumers, users and the environment, and on its efficacy. The identity of the active agent was established in a previous assessment and belongs to a bacterial species whose safety EFSA considers can be assessed using the QPS approach. Thus, the product strain meets the qualifications of this group of bacilli and the additive can be presumed safe for the target species, consumers and environment. Three trials conducted with turkeys showed a significant increase in final body weight in birds given the additive at the proposed dose compared to control animals. Consequently, Calsporin can be considered as efficacious for turkeys for fattening at the dose of 3 x 10 8 CFU/kg feed. Since the efficacy of Calsporin for chickens and turkeys for fattening was established, it can be assumed to be efficacious with chickens reared for laying and turkeys reared for breeding at the proposed doses. As the mechanism of action of the additive can be reasonably assumed to be the same as in the major species, and as the proposed dose is the same, efficacy for the minor avian species can be presumed at the dose of 3 x 10 8 CFU/kg feed. The compatibility of Calsporin was demonstrated with authorised coccidiostats. 1 On request from the European Commission, Question No EFSA-Q , adopted on 5 October Panel members: Gabriele Aquilina, Georges Bories, Paul Brantom, Andrew Chesson, Pier Sandro Cocconcelli, Joop de Knecht, Noël Albert Dierick, Mikolaj Antoni Gralak, Jürgen Gropp, Ingrid Halle, Reinhard Kroker, Lubomir Leng, Sven Lindgren, Anne-Katrine Lundebye Haldorsen, Alberto Mantovani, Miklós Mézes, Derek Renshaw and Maria Saarela. Correspondence: FEEDAP@efsa.europa.eu 3 Acknowledgement: The Panel wishes to thank the members of the Working Group on Micro-organisms including Guido Rychen for the preparation of this opinion. Suggested citation: EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP); Scientific Opinion on the safety and efficacy of Calsporin (Bacillus subtilis) for turkeys for fattening, ducks, geese, pigeons and other game birds for meat production, ducks, geese, pigeons, game birds, ornamental and sporting birds for rearing to point of lay, turkeys reared for breeding and chickens reared for laying.. [13 pp.]. doi: /j.efsa Available online: European Food Safety Authority,

2 KEY WORDS Zootechnical additive, gut flora stabiliser, Calsporin, Bacillus subtilis, micro-organism, QPS, efficacy, turkeys for fattening, chickens reared for laying 2

3 SUMMARY 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 Calsporin (Bacillus subtilis) for chickens, turkeys and minor avian species. Calsporin is a microbial feed additive based on a single strain of Bacillus subtilis. The applicant is seeking authorisation for the use of the product as a zootechnical feed additive (functional group: gut flora stabilisers) for turkeys for fattening and reared for breeding, ducks, geese, pigeons and other game birds for meat production, and ducks, geese, pigeons, game birds, ornamental and sporting birds for rearing to point of lay at a recommended dose of 3 x 10 8 CFU/kg complete feedingstuff, and for chickens reared for laying at the recommended dose of 5 x 10 8 CFU/kg complete feedingstuff. The additive has already been assessed for use as feed additive for chickens for fattening and for weaned piglets by the European Food Safety Authority (EFSA). The identity of the active agent was conclusively established in the previous application and belongs to a bacterial species that is considered to be suitable for the QPS approach to safety assessment. Consequently, the additive can be presumed safe for the target species, consumers and the environment. The extension of use of the additive to the new target species is considered unlikely to introduce hazards for users of the product not already considered in the previous assessments. Three studies were provided with turkeys for fattening which showed that Calsporin has the potential to improve weight gain at the proposed dose of 3 x 10 8 CFU/kg feed. Since the efficacy of Calsporin for chickens (5 x 10 8 CFU/kg feed) and turkeys for fattening (3 x 10 8 CFU/kg feed) has been established, the product can be assumed to be efficacious with chickens reared for laying and turkeys reared for breeding at the same respective doses. Moreover, as the mechanism of action of the additive can be reasonably assumed to be the same as in the major species, efficacy for ducks, geese, pigeons and other game birds for meat production, ducks, geese, pigeons, game birds, ornamental and sporting birds for rearing to point of lay can be presumed at a dose of 3 x 10 8 CFU/kg feed. The compatibility of simultaneous use of Calsporin with the following coccidiostats: diclazuril, halofuginone, decoquinate, nicarbazin, lasalocid A sodium, monensin sodium, robenidine hydrochloride, maduramycin ammonium, narasin, narasin/nicarbazin, semduramycin sodium and salinomycin, was demonstrated. 3

4 TABLE OF CONTENTS Abstract... 1 Key words... 2 Summary... 3 Table of contents... 4 Background... 5 Terms of reference... 5 Assessment Introduction Characterisation Stability and homogeneity Conditions of use Evaluation of the analytical methods by the Community Reference Laboratory (CRL) Efficacy Efficacy for turkeys for fattening Efficacy for chickens reared for laying Efficacy for turkeys reared for breeding Efficacy for ducks, geese, pigeons and other game birds for meat production, ducks, geese, pigeons, game birds, ornamental and sporting birds for rearing to point of lay Compatibility with coccidiostats in poultry feed Post-market monitoring Conclusions Documentation provided to EFSA References

5 BACKGROUND Regulation (EC) No 1831/ 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 the company Calpis Co. Ltd Japan 5 for authorisation of the product Calsporin, Bacillus subtilis, to be used as a feed additive for ducks, geese, pigeons and other game birds for meat production, ducks, geese, pigeons, game birds, ornamental and sporting birds for rearing to point of lay, turkeys reared for breeding and for fattening, and chickens reared for laying (category: zootechnical additives; functional group: gut flora stabilisers) under the conditions mentioned in Table 1. 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. 6 According to Article 8 of that Regulation, 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. The particulars and documents in support of the application were considered valid by EFSA as of 9 June The additive Calsporin is a preparation of Bacillus subtilis C This product is authorised for use with chickens for fattening 7 and weaned piglets. 8 EFSA issued two opinions on the safety and efficacy of Calsporin for chickens for fattening (EFSA, 2006 and 2007), and one on the safety and efficacy of Calsporin for piglets (EFSA, 2010). TERMS OF REFERENCE According to Article 8 of Regulation (EC) No 1831/2003, EFSA shall 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 the efficacy of the product Calsporin (Bacillus subtilis), when used under the conditions described in Table 1. 4 OJ L 268, , p Calpis Co. Ltd Japan, represented in EU by Calpis Co. Ltd. Europe Representative Office, 153 Rue de Courcelles, 75017, Paris Cedex 17, France. 6 EFSA Dossier reference: FAD OJ L 271, , p OJ L 102, , p

6 Table 1: Description and conditions of use of the additive as proposed by the applicant Additive Calsporin Registration number/ec No/No Category of additive 4b1820 Zootechnical feed additive Functional group of additive Gut flora stabiliser (micro-organism) Composition, description A preparation of Bacillus subtilis C-3102 (DSM 15544) Description Chemical Purity criteria formula Minimum of 1.0 x viable Complies with EU feed spores (CFU) hygiene law per gram Method of analysis BS EN 15784:2009 Trade name Calsporin Name of the holder of authorisation Calpis Co. Ltd., Japan, represented in the EU by the Calpis Co. Ltd. Europe Representative Office, Paris ; France Conditions of use Species or category of animal Turkeys for fattening. Maximum Age To slaughter Minimum content Maximum content Withdrawal period CFU/kg of complete feedingstuffs 3 x 10 8 CFU/kg Turkeys for breeding/laying. Minor avian species for fattening. Minor avian species for breeding/laying. Chickens for breeding/laying. To point of lay To slaughter To point of lay To point of lay 3 x 10 8 CFU/kg 3 x 10 8 CFU/kg 3 x 10 8 CFU/kg 5 x 10 8 CFU/kg Other provisions and additional requirements for the labelling Specific conditions or restrictions for use Specific conditions or restrictions for handling Post-market monitoring None R42, potential respiratory sensitiser Breathing protection during handling and safety glasses No specific requirements 6

7 Specific conditions for use in complementary feedingstuffs Dosage used should supply at least minimum recommended CFU/kg final complete feedingstuff. Marker residue Maximum Residue Limit (MRL) Species or category of animal Target tissue(s) or food products Maximum content in tissues

8 ASSESSMENT 1. Introduction Calsporin is a preparation of viable spores of Bacillus subtilis C The applicant has requested an authorisation for use with ducks, geese, pigeons and other game birds for meat production, ducks, geese, pigeons, game birds, ornamental and sporting birds for rearing to point of lay, turkeys reared for fattening and for breeding, and chickens reared for laying, under the category of a zootechnical additive (functional group: gut flora stabilisers). This product is currently authorised for use in chickens for fattening and weaned piglets (see Background). B. subtilis is considered by the European Food Safety Authority (EFSA) to be suitable for the Qualified Presumption of Safety (QPS) approach to safety assessment (EFSA, 2009). In the course of previous assessments the identity of the bacterial strain was conclusively established and the absence of a toxigenic potential demonstrated. In those assessments the susceptibility of the strain to the battery of antibiotics recommended by the Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) was also demonstrated (EFSA, 2010). The strain was thus considered to satisfy the qualifications specified for the species and so can be treated as meeting the requirements of the QPS approach. Consequently, no further assessment of safety for the target species, the consumer and the environment is required. In addition, the use of the additive with ducks, geese, pigeons and other game birds for meat production, ducks, geese, pigeons, game birds, ornamental and sporting birds for rearing to point of lay, turkeys reared for breeding and for fattening, and chickens reared for laying is considered unlikely to introduce hazards for users of the product not already considered as part of the first assessment (EFSA, 2006). Consequently, in the present opinion the FEEDAP Panel has focused only on the data specific to the use of the additive with the new target species and with the compatibility of Calsporin with coccidiostats. 2. Characterisation The active agent in the additive consists of viable endospores of a single strain of B. subtilis originally isolated from soil in Japan and deposited in the German Collection of Microorganisms and Cell Cultures (DSMZ) with the accession number DSM Stability and homogeneity The stability of three batches of the additive was established under two storage conditions (25 o C/60% RH and 40 o C/75% RH) and the shelf life shown to be at least of 39 and eight months, respectively. 10 No or a small reduction in microbial counts (minimum B. subtilis levels 91 % of the time zero value) was seen in three batches of Calsporin when mixed with a premixture for chickens after six months of storage at 25 o C/60% RH. The reduction over the same period at 40 o C/75% RH was higher (12 21 %). 11 The survival of the active agent (three batches) during pelleting at o C and subsequent storage was studied in turkey feeds (starter, grower, finisher). Pelleting resulted in a reduction of % in the B. subtilis counts, while recovery after three months of storage under practical farm conditions was 100 % of the post-pelleting values. 12 A similar test was done in feed for chickens for fattening (three batches) at 70 ºC, which resulted in % losses of B. subtilis. 13 Storage of these pelleted feeds at 25 o C/60% RH and 40 o C/75% RH for three months showed a survival rate of % and 33 9 Technical dossier/section II/Annex II_2_1_2a. 10 Technical dossier/section II/Annex II_4_1b. 11 Technical dossier/section II/Annex II_4_1c. 12 Technical dossier/section II/Annex II_4_1d. 13 Technical dossier/section II/Annex II_4_1f. 8

9 40 % of the initial post-pelleting values, respectively. 14 In another study, pelleting of feed for chickens for fattening at 70, 80 and 90 ºC resulted in recoveries of %, % and %, respectively. 15 In summary, the data suggest losses during pelleting of approximately 30 % of viable spores and further susceptibility of the additive in those feeds during storage. The capacity of the product to homogeneously distribute in feed has been previously demonstrated for chickens feed (EFSA, 2006). The FEEDAP Panel considers these data to be applicable for turkey feed Conditions of use The product is intended for use in feed for ducks, geese, pigeons and other game birds for meat production, ducks, geese, pigeons, game birds, ornamental and sporting birds for rearing to point of lay and turkeys reared for breeding and for fattening at a dose of 3 x 10 8 CFU/kg complete feedingstuffs, and for chickens reared for laying at a dose of 5 x 10 8 CFU/kg complete feedingstuffs Evaluation of the analytical methods by the Community Reference Laboratory (CRL) The CRL considered that the conclusions and recommendations reached in the previous assessment are valid and applicable for the current application. 3. Efficacy 3.1. Efficacy for turkeys for fattening Three efficacy studies with turkeys for fattening with a similar experimental design, carried out in two different Member States, have been reported. Trial 1 In the first trial, a total of 799 one-day-old female turkeys for fattening (BUT Nº 9) were distributed in 24 replicates of 33 or 34 birds per replicate, adjusted (based on weight) to 25 birds per pen after 28 days. 16 The turkeys were randomly allocated to two experimental treatments (control and Calsporin ) in a block design using two identical but separate rooms in order to prevent cross-contamination. Birds were fed typical commercial turkey diets in mash or pellets, depending on the age of the birds. Calsporin was added to basal diets providing 3 x 10 8 CFU/kg feed (confirmed by analysis), otherwise the diets were identical in composition and nutrient homogeneity. The study lasted 84 days. At the end of the study, birds receiving Calsporin were heavier and consumed more feed (P < 0.05) than the birds of the control group (Table 2). Feed to gain ratio was not significantly different between the treatments. Overall mortality was low (< 4 %) and did not differ between treatments. Trial 2 In the second trial, Calsporin was added to a basal diet based on wheat and soybean meal at 30 mg/kg, providing 3 x 10 8 CFU/kg feed (confirmed by analysis). 17 Two treatments were established (control and Calsporin ) each of which was fed to 32 replicates of 15 one-day-old birds (960 male commercial BUT 8 turkeys in total) for 84 days. The diets (starter, grower and finisher) were provided for ad libitum access. The overall rate of mortality/culling was 2.1 %. Over the entire period, the birds fed the diet containing Calsporin were significantly heavier, consumed significantly more feed, and had significantly better feed to gain ratio than birds fed the control diet (Table 2). 14 Technical dossier/section II/Annex II_4_1f. 15 Technical dossier/section II/Annex II_4_1 e. 16 Technical dossier/section IV/Annex II_3a. 17 Technical dossier/section IV/Annex II_3b. 9

10 Trial 3 In the third trial, 400 one-day-old hybrid Large White male poults were randomly allotted into 16 pens (25 poults per pen) blocked by treatment. 18 The treatments consisted in a basal diet and the same diet containing Calsporin at 3 x 10 8 CFU/kg feed (doses verified by analysis). Lasalocid A sodium was included in all diets (90 mg/kg feed). The trial lasted 126 days. Individual birds weight and pen feed consumption data were collected. Feed to gain ratio was adjusted to account for mortality. Over the entire period, the birds fed diets containing Calsporin were significantly heavier than birds in the control group. No differences in feed to gain ratio (Table 2) or mortality were observed. Table 2: Summary of the results of efficacy trials of Calsporin with turkeys Trial (Duration in days) 1 (84) 2 (84) 3 (126) a,b 1 Total animals (Replicates/treatment X animals/replicate) 600 (12 X 25) 960 (32 X 15) 400 (8 X 25) Treatment (CFU/kg feed) Total weight gain 1 (kg) Feed to gain ratio 0 3x a 6.7 b a 2.05 a 3x b 2.02 b a x b 2.42 Means within a treatment not sharing a common superscript differ significantly P < Final Body Weight in trial 1. The three trials provided showed that turkeys receiving Calsporin at the recommended dose were heavier than control animals. An improved feed to gain ratio was also observed in one trial Efficacy for chickens reared for laying The efficacy for chickens for fattening has been established in previous assessments (EFSA, 2006 and 2007). Since the applicant proposes the use of the same dose (5 x 10 8 CFU/kg complete feedingstuffs) with chickens reared for laying, efficacy for this animal category can be presumed Efficacy for turkeys reared for breeding The efficacy for turkeys for fattening was established in the studies described above. Since the applicant proposes the use of the same dose (3 x 10 8 CFU/kg complete feedingstuffs) with turkeys reared for breeding, efficacy for this animal category can be presumed Efficacy for ducks, geese, pigeons and other game birds for meat production, ducks, geese, pigeons, game birds, ornamental and sporting birds for rearing to point of lay The efficacy for turkeys for fattening was established in the studies described above. Since the applicant proposes the use of the same dose (3 x 10 8 CFU/kg complete feedingstuffs) in these minor avian species, and as the mechanism of action of the additive can be reasonably assumed to be the same as in the major species, efficacy for ducks, geese, pigeons and other game birds for meat production, ducks, geese, pigeons, game birds, ornamental and sporting birds for rearing to point of lay can be presumed. 4. Compatibility with coccidiostats in poultry feed The compatibility of Calsporin with various coccidiostats was studied using the stepwise approach proposed by the FEEDAP Panel (EFSA, 2008). Five chemical coccidiostats (decoquinate, robenidine hydrochloride, diclazuril, nicarbazin and halofuginone) and six ionophores (lasalocid, narasin, 18 Technical dossier/section IV/Bibliography/Blair_et_al_

11 salinomycin, maduramycin ammonium, monensin sodium, and semduramycin sodium) were evaluated for their minimum inhibitory concentrations (MIC) against Bacillus subtilis C The MIC values of diclazuril (> 4 µg/ml), halofuginone (> 16 µg/ml), nicarbazin (> 512 µg/ml) and decoquinate ( 256 µg/ml) were higher than four times the maximum authorised concentrations of these coccidiostats in feed. Therefore, compatibility is assumed with these four coccidiostats. Compatibility could not be established in vitro for salinomycin sodium, monensin sodium, semduramycin sodium, lasalocid A sodium, maduramycin ammonium, robenidine hydrochloride and narasin. Therefore, to demonstrate the compatibility with those coccidiostats, an in vivo trial was performed. 20 Three hundred and sixty one-day-old Ross 308 male chicks were allocated at random to nine treatments (T1 to T9, Table 3), replicated six times each (four to nine birds per cage). Experimental feeds and water were provided for ad libitum access from arrival until gut sampling. Feeds (T2 to T9) were supplemented with Calsporin at the recommended dose, and with one of the coccidiostats each at the maximum authorised (or higher) dose (T3 to T9, Table 9). The duration of the study was 35 days. Birds were sacrificed from week 3 to 5 and fresh gut contents taken. At least six samples of approximately 15 g of caecal/large bowel contents were obtained per treatment (nine samples for T1 and T2). Differences in total numbers of Bacillus subtilis spores or vegetative cells between each coccidiostat treatment and the positive control did not reach statistical significance (Table 3). Table 3: Viable spores and vegetative cells of Bacillus subtilis in gut contents of chickens for fattening Treatment Spores Vegetative cells Log 10 CFU/g Log 10 CFU/g T1 basal diet ND* ND T2 Calsporin T3 Calsporin + lasalocid A sodium 125 mg/kg T4 Calsporin + monensin sodium 125 mg/kg T5 Calsporin + robenidine hydrochloride 36 mg/kg T6 Calsporin + maduramycin ammonium 5 mg/kg T7 Calsporin + narasin 70 mg/kg T8 Calsporin + narasin/nicarbazin 100 mg/kg (50+50) T9 Calsporin + semduramycin sodium 25 mg/kg * ND: not detected; below the detection level 3.0. The in vivo compatibility of Calsporin with lasalocid A sodium, monensin sodium, robenidine hydrochloride, maduramycin ammonium, narasin, narasin/nicarbazin and semduramycin sodium was demonstrated in the present trial. The compatibility of Calsporin with salinomycin sodium was tested in a second in vivo study. 21 The experiment involved one-day-old Ross chicks and consisted of three treatments (T1 basal diet, T2 Calsporin, T3 Calsporin + salinomycin sodium 70 mg/kg) with eight replicates each (900 birds/replicate). Calsporin with or without the coccidiostat was added at the recommended dose to the starter (from day 0 to day 18) and grower (from day 18 to day 35) diets. Faecal samples were collected from four pens per treatment at days 13, 21 and 31 for the analysis of B. subtilis vegetative cells and spores (T2 and T3). The results are shown in Table 4. The levels of B. subtilis spores were the same in T2 and T3, whereas the levels of vegetative cells were Log-value lower in T3 compared to T2 (depending on a time point) (Table 4). Since the difference between T2 and T3 vegetative cells was less than one Log 10 value, the in vivo compatibility of Calsporin with salinomycin was demonstrated. 19 Technical dossier/section II/Annex II_4_4a. 20 Technical dossier/section II/Annex II_4_4b. 21 Technical dossier/section II/Annex II_4_4c. 11

12 Table 4: Viable spores and vegetative cells of Bacillus subtilis in the faeces of chickens for fattening at different time points Treatment Spores Vegetative cells (Log 10 CFU/g) (Log 10 CFU/g) 13d 21 d 31 d 13 d 21 d 31 d T1 basal diet ND* ND 4.1 T2 Calsporin T3 Calsporin +salinomycin sodium 70 mg/kg * Not detected; below the detection level. In conclusion, the compatibility of Calsporin was demonstrated with diclazuril, halofuginone, decoquinate, nicarbazin, lasalocid A sodium, monensin sodium, robenidine hydrochloride, maduramycin ammonium, narasin, narasin/nicarbazin, semduramycin sodium and salinomycin sodium. 5. 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 Regulation 22 and Good Manufacturing Practice. CONCLUSIONS The active agent Bacillus subtilis is considered by EFSA to be suitable for the QPS approach to safety assessment. As the identity of the active agent has been established and the susceptibility to antibiotics and lack of toxigenic potential have been demonstrated, the use of Calsporin can be presumed safe for the target species for which an authorisation is being sought. Evidence has been provided that Calsporin has a potential to improve weight gain when fed to turkeys for fattening at the proposed dose of 3 x 10 8 CFU/kg complete feedingstuffs. Since the efficacy of Calsporin for chickens (5 x 10 8 CFU/kg complete feedingstuffs) and turkeys for fattening (3 x 10 8 CFU/kg complete feedingstuffs) has been established, the product can be assumed to be efficacious with chickens reared for laying and turkeys reared for breeding at the same respective doses. Moreover, as the mechanism of action of the additive can be reasonably assumed to be the same as in the major species, efficacy for ducks, geese, pigeons and other game birds for meat production, ducks, geese, pigeons, game birds, ornamental and sporting birds for rearing to point of lay can be presumed at a dose of 3 x 10 8 CFU/kg complete feedingstuffs. The compatibility of Calsporin was demonstrated with diclazuril, halofuginone, decoquinate, nicarbazin, lasalocid A sodium, monensin sodium, robenidine hydrochloride, maduramycin ammonium, narasin, narasin/nicarbazin, semduramycin sodium and salinomycin. DOCUMENTATION PROVIDED TO EFSA 1. Application for extension of the authorisation of Calsporin (Bacillus subtilis C-3102, EU No 4b1820) as a feed additive in the EU as per Regulation (EC) No 1831/2003. April Submitted by Pen & Tec Consulting. 2. Comments from Member States received through the ScienceNet. 22 OJ L 35, , p

13 REFERENCES EFSA (European Food Safety Authority), Scientific Opinion of the Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) on the safety and efficacy of the product Calsporin, a preparation of Bacillus subtilis, as a feed additive for chickens for fattening in accordance with Regulation (EC) No 1831/2003. The EFSA Journal, 336, EFSA (European Food Safety Authority), Scientific Opinion of the Panel on additives and products or substances used in animal feed (FEEDAP) on the safety and efficacy of the product Calsporin, a preparation of Bacillus subtilis, as a feed additive for chickens for fattening in accordance with Regulation (EC) No 1831/2003. The EFSA Journal, 543, 1-8. EFSA (European Food Safety Authority), Technical guidance prepared by the Panel on Additives and Products or Substances Used in Animal Feed (FEEDAP) on the compatibility of zootechnical microbial additives with other additives showing antimicrobial activity. The EFSA Journal, 658, 1-5. EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP), Scientific Opinion on the maintenance of the list of QPS microorganisms intentionally added to food or feed. EFSA Journal, 7(12):1431. EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP), Scientific Opinion on the safety and efficacy of Calsporin (Bacillus subtilis) as a feed additive for piglets on request from the European Commission. EFSA Journal, 8(1):