Soil microbial community responses to different sugarcane management strategies as revealed by 16S metagenomics

XXX Congress of the ISSCT (International Society of Sugar Cane Technologists), Tucumán, from 31 August to 8 September 2019Sugarcane cultivation in Argentina is distributed in three geographic regions: Tucumán, Northern (Salta and Jujuy) and Littoral (Santa Fe and Misiones), covering about 376,223 ha. Tucumán has traditionally been the most important region with 68% of the total production. Since new agricultural techniques, such as green-cane harvesting and sugarcane crop rotation with soybean, were implemented in the last decade, changes in the agroecosystem of sugarcane, specific pathosystems and epidemiological parameters have been observed. A 16S metagenomics approach to investigate total bacterial communities associated with sugarcane rhizospheric soil when soybean was the predecessor crop in a cultivation area with a high incidence of red stripe disease was applied; soil from sugarcane monoculture was also included. Two commercial sugarcane cultivars (Saccharum spp. hybrids) with differential responses to red stripe infection (tolerant and susceptible) were evaluated. Sampling was carried out in 2013-2014 and in 2014-2015 (first and second ratoon, respectively) at 30, 90 and 180 days after harvest. Total soil DNA was obtained using FastPrep® technology. The 16S RNA gene (variable region V3-V4) was sequenced using a MiSeq platform Illumina. Taxonomic assignment revealed Bacillales, Rhyzobiales, Rhodospirilliales, Xanthomonadales and Acidobacteriales among the most abundant orders in all samples. Soil samples from sugarcane without soybean rotation showed a marked decrease in Bacillaes, Rhizobiales and Sphingomonadales. Cluster analysis grouped together samples from the tolerant genotype, while those from the susceptible genotype formed two subgroups that were distinguished according to sampling time after harvest. The analysis showed that samples from sugarcane under monoculture were grouped distant to the rest of the samples showing different microbiota composition. The sugarcane rhizosphere microbiome and its biotechnological potential open a new opportunity in the concept of sustainable crop management. The data contribute significant knowledge about the microbial diversity in agricultural ecosystems.El cultivo de caña de azúcar en Argentina se distribuye en tres regiones geográficas: Tucumán, Norte (Salta y Jujuy) y Litoral (Santa Fe y Misiones), que cubren alrededor de 376,223 ha. Tucumán ha sido tradicionalmente la región más importante con el 68% de la producción total. En la última década, como consecuencia de los cambios en el manejo como la rotación con soja y la cosecha en verde, se han observado cambios en el agroecosistema de la caña de azúcar, en patosistemas específicos y parámetros epidemiológicos. En el presente estudio, se aplicó un enfoque metagenómica de gen 16S ARNr para investigar las comunidades bacterianas asociadas con el suelo rizosférico de caña de azúcar en un sistema con soja como cultivo predecesor en un área de cultivo con una alta incidencia de estría roja. A fines comparativos, se incluyó en el estudio muestras de suelo monocultivo de caña de azúcar. Se evaluaron dos cultivares comerciales de caña de azúcar (Saccharum spp. híbridos) con respuestas diferenciales a la infección de estría roja (tolerante y susceptible). A partir del ADN total del suelo de muestras obtenidas en las campañas 2013-2014 y en 2014-2015 (soca 1 y 2, respectivamente) a los 30, 90 y 180 días después de la cosecha, se amplificó la región variable V3-V4 del 16S rRNA y se secuenció utilizando una plataforma MiSeq Illumina. La asignación taxonómica de las secuencias, reveló Bacillales, Rhyzobiales, Rhodospirilliales, Xanthomonadales y Acidobacteriales entre las órdenes más abundantes en todas las muestras. Las muestras de suelo de caña de azúcar sin rotación con soja mostraron una marcada disminución en Bacillaes, Rhizobiales y Sphingomonadales. El análisis de conglomerados agrupó las muestras del genotipo tolerante, mientras que las del genotipo susceptible formaron dos subgrupos que se distinguieron según el tiempo de muestreo después de la cosecha. El análisis mostró además que las muestras de caña de azúcar provenientes de un sistema de monocultivo se agruparon distantes del resto de las muestras evidenciando las diferencias en la composición de microbiota. El microbioma de la rizosfera de la caña de azúcar y su potencial biotecnológico abren una nueva oportunidad en el concepto de manejo sostenible de cultivos. Los datos aportan un conocimiento significativo sobre la diversidad microbiana en los ecosistemas agrícolas.La culture de la canne à sucre en Argentine est répartie dans trois régions géographiques: Tucumán, le Nord (Salta et Jujuy) et le Littoral (Santa Fe et Misiones), couvrant environ 376 223 ha. Tucumán est traditionnellement la région la plus importante avec 68% de la production totale. Depuis que de nouvelles techniques agricoles, telles que la récolte de la canne verte et la rotation de la canne à sucre avec le soja, ont été mises en place au cours de la dernière décennie, des changements dans l’écosystème de la canne à sucre, des pathosystèmes spécifiques et des paramètres épidémiologiques ont été observés. Une approche métagénomique 16S pour étudier les communautés bactériennes totales associées au sol rhizosphérique de la canne à sucre lorsque le soja était la culture précédente dans une zone de culture présentant une incidence élevée de maladie à rayures rouges a été appliquée; le sol issu de la monoculture de canne à sucre a également été inclus. Deux cultivars commerciaux de la canne à sucre (hybrides Saccharum spp.) avec des réponses différentielles à l'infection à rayures rouges (tolérants et sensibles) ont été évalués. L'échantillonnage a été réalisé en 2013-2014 et en 2014-2015 (première et deuxième repousses, respectivement) à 30, 90 et 180 jours après la récolte. L'ADN total du sol a été obtenu en utilisant la technologie FastPrep®. Le gène de l'ARN 16S (région variable V3-V4) a été séquence en utilisant une plate-forme MiSeq Illumina. L'analyse taxonomique a révélé que Bacillales, Rhyzobiales, Rhodospirilliales, Xanthomonadales et Acidobacteriales étaient parmi les ordres les plus abondants dans tous les échantillons. Des échantillons de sol de canne à sucre sans rotation de soja ont montré une diminution marquée des bacilles, des rhizobiales et des sphingomonadales. L'analyse a regroupé des échantillons du génotype tolérant, tandis que ceux du génotype sensible ont formé deux sous-groupes qui ont été distingués en fonction de la période d'échantillonnage après la récolte. L'analyse a montré que les échantillons provenant de la canne à sucre en monoculture étaient regroupés à distance du reste des échantillons présentant une composition de microbiote différente. Le microbiome de la rhizosphère de canne à sucre et son potentiel biotechnologique ouvrent une nouvelle opportunité dans le concept de gestion durable des cultures. Les données fournissent des connaissances significatives sur la diversité microbienne dans les écosystèmes agricoles.EEA FamailláFil: Fontana, Paola Daniela. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Famaillá; ArgentinaFil: Orru, L. Council for Agricultural Research and Economics (CREA). Research Centre for Genomics & Bioinformatics; ItaliaFil: Fontana, Cecilia Alejandra. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Famaillá; ArgentinaFil: Cocconcelli, P.S. Università Cattolica del Sacro Cuore. Dipartimento di Scienze e Tecnologie Alimentari per una filiera agro-alimentare Sostenibile (DISTAS); ItaliaFil: Vignolo, Graciela Margarita. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Tucumán. Centro de Referencia Para Lactobacilos; ArgentinaFil: Salazar, Sergio Miguel. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Famaillá; Argentin

Multilocus Sequence Analysis highlights genetic diversity of Acidovorax avenae strains associated with sugarcane red stripe

XXX Congress of the ISSCT (International Society of Sugar Cane Technologists), Tucumán, from 31 August to 8 September 2019Pathogenic species of Acidovorax cause economically important diseases in monocotyledonous and dicotyledonous crops, including sugarcane, corn, rice, oats, millet, foxtail, watermelon and orchids. Sugarcane red stripe, caused by Acidovorax avenae, is present in the main production areas around the world. In Argentina, red stripe affects about 30% of stalks milled with important economic losses when severe infections occur. MLST was used to explore the genetic diversity of this bacterium associated with red stripe in Argentina, as well as their phylogenetic relationships. The MLST analysis included sequences from a total of 118 Acidovorax, 15 A. avenae strains isolated from Argentina sugarcane production areas, A. citrulli (93) from melon and watermelon, A. avenae (9) from rice, millet, corn, vasey grass and sorghum, and A. oryzae (1) from rice. MLST analysis revealed five novel sequence types (STs) for the sugarcane A. avenae strains, constituting a clonal complex with a common and close origin. When genetic relationships with other Acidovorax were explored, sugarcane strains were related to A. avenae from other hosts and more distantly to A. citrulli. Signals of frequent recombination in several lineages of A. avenae were detected and we observed that A. oryzae is closely related to A. avenae strains. This study provides valuable data in the field of epiphytological and evolutionary investigations of A. avenae strains causing sugarcane red stripe. Knowledge of the genetic diversity and host-strain specificity are important to select the genotypes with the best response to red stripe disease.Las especies fitopatógenas de Acidovorax causan enfermedades en cultivos tanto monocotiledóneos y dicotiledóneos, que incluyen caña de azúcar, maíz, arroz, avena, mijo, sandía y orquídeas. La estría roja de caña de azúcar, causada por Acidovorax avenae, está presente en las principales áreas de producción del mundo. En Argentina, esta enfermedad llegó a afectar hasta un 30% de tallos molibles en infecciones severas con importantes pérdidas económicas. Para explorar la diversidad genética de esta bacteria, así como sus relaciones filogenéticas, se aplicó un análisis MLST. El MLST incluyó un total de 118 secuencias de cepas de Acidovorax, 15 A. avenae aisladas de diferentes áreas de producción de caña de azúcar de Argentina, A. citrulli (93) de melón y sandía, A. avenae (9) de arroz, mijo, maíz, pasto vasey y sorgo y A. oryzae (1) de arroz. El análisis de MLST reveló cinco nuevos tipos de secuencia (ST) para las cepas de caña de azúcar A. avenae, que constituyen un complejo clonal con un origen común y cercano. Cuando se investigó la relación genética con otras Acidovorax, las cepas de caña de azúcar se mostraron cercanas con A. avenae de otros huéspedes, pero más distante de A. citrulli. Se evidenciaron señales de recombinación frecuente en varios linajes de A. avenae y observamos que A. oryzae está estrechamente relacionada con las cepas de A. avenae. Este estudio proporciona datos valiosos en el campo de las investigaciones epifitológicas y evolutivas de las cepas de A. avenae que causan estría roja en caña de azúcar. El conocimiento de la diversidad genética y la especificidad cepa-hospedante son importantes para seleccionar genotipos con la mejor respuesta frente a los biotipos más virulentos y predominantes en la región.Les espèces pathogènes d'Acidovorax sont responsables de maladies importantes sur le plan économique dans les cultures monocotylédones et dicotylédones, notamment la canne à sucre, le maïs, le riz, l'avoine, le millet, la sétaire, la pastèque et l’orchidée. Les rayures rouges de la canne à sucre, causée par Acidovorax avena, est présente dans les principales zones de production du monde. En Argentine, les rayures rouges touchent environ 30% des tiges usinables, entraînant d'importantes pertes économiques en cas d'infection grave. Le MLST a été utilisé pour explorer la diversité génétique de cette bactérie associée aux rayures rouges en Argentine, ainsi que leurs relations phylo-génétiques. L'analyse MLST comprenait des séquences provenant d'un total de 118 souches Acidovorax, 15 souches d'A. avenae isolées des zones de production de canne à sucre de l’Argentine, A. citrulli (93) du melon et de pastèque, A. avenae (9) du riz, le mil, le maïs, l’herbe de vasey et le sorgho et A. oryzae (1) obtenue du riz. L'analyse MLST a révélé cinq nouveaux types de séquence (ST) pour les souches de canne à sucre d’A. avenae, constituant un complexe clonal d'origine commune et proche. Lorsque les relations génétiques avec d'autres Acidovorax ont été explorées, les souches provenant de la canne à sucre étaient apparentées à A. avenae provenant d'autres hôtes et plus lointainement à A. citrulli. Des signaux de recombinaison fréquente dans plusieurs lignées d'A. avenae ont été détectés et nous avons observé qu'A. oryzae est étroitement apparenté aux souches d'A. avenae. Cette étude fournit des données précieuses dans le domaine des études épiphytologiques et évolutives des souches d'A. avenae provoquant les rayures rouges de la canne à sucre. La connaissance de la diversité génétique et de la spécificité souche-hôte est importante pour sélectionner les génotypes présentant la meilleure résistance à la maladie des rayures rouges.EEA FamailláFil: Fontana, Paola Daniela. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Famaillá; ArgentinaFil: Tomasini, Nicolás. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Salta. Instituto de Patología Experimental; ArgentinaFil: Tomasini, Nicolás. Universidad Nacional de Salta. Instituto de Patología Experimental; ArgentinaFil: Fontana, Cecilia Alejandra. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Famaillá; ArgentinaFil: Di Pauli, Valentina. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Famaillá; ArgentinaFil: Cocconcelli, P.S. Università Cattolica del Sacro Cuore. Dipartimento di Scienze e Tecnologie Alimentari per una filiera agro-alimentare Sostenibile (DISTAS); ItaliaFil: Vignolo, Graciela Margarita. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Tucumán. Centro de Referencia Para Lactobacilos; ArgentinaFil: Salazar, Sergio Miguel. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Famaillá; Argentin

Scientific Opinion on the safety and efficacy of Urea for ruminants

Urea supplementation to feed for ruminants provides non-protein nitrogen for microbial protein synthesis in the rumen and thus in part replaces other dietary protein sources. Urea supplementation of feed for ruminants at doses up to 1 % of complete feed DM (corresponding to 0.3 g/kg bw/day) is considered safe when given to animals with a well adapted ruminal microbiota and fed diets rich in easily digestible carbohydrates. Based on the metabolic fate of urea in ruminants, the use of urea in ruminant nutrition does not raise any concern for consumers\u2019 safety. Urea is considered to be non irritant to skin and eyes and its topical use suggests that it is not a dermal sensitiser. The risk of exposure by inhalation would be low. The substitution of protein by urea in well balanced feed for ruminants would not result in an increased environmental nitrogen load. Urea is an effective source of non-protein nitrogen substituting for dietary protein in ruminants

Safety and efficacy of B‐Act® (Bacillus licheniformis DSM 28710) for chickens for fattening and chickens reared for laying

The additive B-Act\uae is a preparation containing viable spores of a strain of Bacillus licheniformis. The additive is intended for use in feed for chickens for fattening and chickens reared for laying at the proposed dose of 1.6 9 109 colony-forming unit (CFU)/kg complete feedingstuffs. B. licheniformis is considered by EFSA to be suitable for the qualified presumption of safety approach for establishing safety. As the identity of the active agent was established and the lack of toxigenic potential and resistance to antibiotics of human or veterinary clinical significance were demonstrated, the additive is presumed safe for the target species, consumers and the environment. In the absence of data, no conclusion can be drawn on the skin/eye irritation or skin sensitisation potential. The dustiness of the preparations tested indicated a potential for users to be exposed via inhalation. B-Act\uae should be considered to have the potential to be a respiratory sensitiser. B-Act\uae at the recommended dose 1.6 9 109 CFU/kg feed has some potential to improve the feed to gain ratio of chickens for fattening. This conclusion can be extended to chickens reared for laying when used at the same dose. B. licheniformis DSM 28710 is compatible with decoquinate, diclazuril, halofuginone, nicarbazin, robenidine hydrochloride, lasalocid A sodium, maduramicin ammonium, monensin sodium, narasin and salinomycin sodium

Safety and efficacy of Taminizer D (dimethylglycine sodium salt) as a feed additive for chickens for fattening

Following a request from the European Commission, EFSA was asked to deliver a scientific opinion on the safety and efficacy of Taminizer D (dimethylglycine sodium salt) as a feed additive for chickens for fattening, based on a dossier submitted for the modification of the terms of authorisation of the additive. The product is authorised in the European Union for chickens for fattening at the maximum content of 1,000 mg/kg complete feedingstuffs. The applicant proposed the introduction of an additional manufacturing process, which introduces an impurity (dimethylamino-ethanol (DMAE)) in the additive at concentrations up to 0.09%. The EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) considered that the proposed modification would not substantially affect the previous assessment as related to the safety of the environment and the efficacy of the product. Since the safety of the active substance was established, the current assessment has dealt with the impurity DMAE. Considering the toxicological profile of DMAE, the estimated intake by the target animal and consumers, and making use of the Threshold of Toxicological Concern (TTC) approach, the Panel concluded that Taminizer D, manufactured by the DMAE route, is safe for both chickens for fattening and consumers, up to the maximum level of 1,000 mg/kg feed. The FEEDAP Panel extends its conclusions about Taminizer D produced by the original method to cover also Taminizer D produced by the new DMAE method. There is minimal risk to users from dust produced as a result of normal handling of the additive. Taminizer D is not irritant to skin but may be irritant to eyes; it is regarded as a potential skin sensitiser. The FEEDAP Panel recommended to set a specification for the DMAE content in the additive

Safety and efficacy of Kelforce \uae (l-glutamic acid, N,N-diacetic acid, tetrasodium salt (GLDA-Na 4 )) as a feed additive for chickens for fattening

l-Glutamic acid, N,N-diacetic acid, tetrasodium salt (GLDA-Na 4 ) (Kelforce \uae ) is sought to be used as a zootechnical feed additive in chickens for fattening to improve the absorption of zinc from feed, reducing zinc emissions through manure and thus, affecting favourably the environment. The product has not been authorised in the European Union as a feed additive. Kelforce \uae is intended to be marketed as a liquid and solid formulation, containing 65 47% and 65 30% of GLDA-Na 4 , respectively. Kelforce \uae is safe for chickens for fattening at the maximum level of 1,000 mg GLDA-Na 4 /kg complete feed. Based on the toxicological profile of GLDA-Na 4 and the consumer exposure to GLDA-Na 4 and to nitrilotriacetic acid trisodium salt (NTA-Na 3 ; an impurity of the additive), the use of Kelforce \uae at the maximum proposed level in feed of chickens for fattening is of no concern for consumer safety. Due to its low inhalation toxicity, the exposure to GLDA-Na 4 is unlikely to pose a risk by inhalation. However, owing to the high-dusting potential of the solid formulation, a risk from such high level of dust, even if toxicologically inert, cannot be excluded. Kelforce \uae is not a skin/eye irritant or skin sensitiser. No risks for the terrestrial compartment were identified at the maximum use level of the additive. Risks for the aquatic compartment cannot be excluded based on the secondary effect of the additive on green algae. In the absence of data, the Panel cannot conclude on the safety for the sediment compartment or the possible ground water contamination. The risk of bioaccumulation and secondary poisoning caused by the additive is considered very low. Owing to the inconsistent and conflicting results from the studies assessed, the Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) cannot conclude on the efficacy of the additive. The Panel made a recommendation regarding the levels of formaldehyde and cyanide in the active substance

Safety and efficacy of Axtra® PHY 20000 TPT2 (6‐phytase) as a feed additive for poultry and porcine species

Axtra\uae PHY 20000 TPT2 is a solid preparation that contains a 6-phytase produced with a genetically modified strain of Trichoderma reesei. The production strain and its recombinant DNA were not detected in Axtra\uae PHY 20000 TPT2. From the results obtained in tolerance studies, the EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) concluded that the additive is safe for the target species at 2,000 FTU/kg feed. The studies provided to address the safety for the consumer were performed with the fermentation product that is used to formulate the additive and the results do not indicate any reason for concern for consumer safety arising from the use of the product as a feed additive. The studies provided to address the safety for the user were performed with the fermentation product that is used to formulate the additive and have been assessed in a previous opinion. Considering the results of those studies and the substances used during the formulation of Axtra\uae PHY 20000 TPT2, this formulation is not considered a dermal sensitiser. However, it should be considered a potential irritant to skin, eyes and the respiratory tract, and owing to the nature of the active substance, it should be considered a potential respiratory sensitiser. However, the exposure by inhalation is expected to be negligible. No risks to the environment are expected from the use of Axtra\uae PHY 20000 TPT2 as a feed additive. Based on the results of efficacy studies, the Panel concluded that the additive has the potential to be efficacious at 250 FTU/kg feed

Assessment of the application for renewal of the authorisation of Natuphos (3-phytase) as a feed additive for poultry and pigs

Natuphos \uae is a feed additive that contains 3-phytase which is produced \u25a0\u25a0\u25a0\u25a0\u25a0 The product is currently authorised for use as a feed additive in chickens for fattening, piglets (weaned) and pigs for fattening, laying hens and turkeys for fattening, ducks, sows, all minor avian species other than ducks and ornamental birds. This scientific opinion concerns the renewal of the authorisation of this additive for those species. The application also included chickens reared for laying/breeding, turkeys reared for breeding and breeding hens. The applicant provided evidence that the additive in the market complies with the conditions of authorisation. According to the information provided by the applicant, no new evidence has been identified that would make the Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) reconsider the previous conclusions regarding the safety for the target species, consumer and environment under the authorised conditions of use. The additive is a respiratory sensitiser and a potential skin sensitiser. The present application for renewal of the authorisation does not include a proposal for amending or supplementing the conditions of the original authorisation that would have an impact on the efficacy of the additive. Therefore, there was no need for assessing the efficacy of the additive in for those species for which an authorisation exists. The Panel also considered that the additive is safe and has a potential to be efficacious in chickens reared for laying/breeding, turkeys reared for breeding, breeding hens and suckling piglets at the corresponding recommended doses

Safety and efficacy of fumonisin esterase from Komagataella phaffii DSM 32159 as a technological feed additive for pigs and poultry

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

Safety and efficacy of TechnoSpore\uae (Bacillus coagulans DSM 32016) for piglets, other growing Suidae, chickens for fattening, other poultry for fattening and ornamental birds

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 TechnoSpore\uae (Bacillus coagulans DSM 32016), when used as a zootechnical additive for piglets (suckling and weaned), other growing Suidae, chickens for fattening, other poultry for fattening and ornamental birds. The bacterial species present in the additive is considered suitable for the qualified presumption of safety approach to safety assessment. The identity of the active agent was established and the lack of toxigenic potential confirmed. B. coagulans DSM 32016 did not show resistance to antibiotics of human and veterinary importance, and therefore, was presumed safe for the target species, consumers of products derived from animals Fed the additive and the environment. Since the other components of the additive did not give rise to concerns, TechnoSpore\uae was also considered safe for the target species, consumer and the environment. The additive is not a skin/eye irritant or a skin sensitiser but should be considered a respiratory sensitiser. TechnoSpore\uae showed the potential to be efficacious as a zootechnical additive in weaned piglets and chickens for fattening at 1  7 109 CFU/kg complete feed. This conclusion was extended to suckling piglets and extrapolated to other growing Suidae at the same physiological stage and to other birds for fattening and ornamental birds at the same use level. B. coagulans DSM 32016 included in Technospore\uae is compatible with halofuginone and diclazuril. The Panel could not conclude on the compatibility of the additive with monensin sodium, decoquinate, robenidine hydrochloride, lasalocid sodium, narasin, salinomycin sodium, maduramicin ammonium, nicarbazin and narasin/nicarbazin