Safety of concentrated l‐lysine (base), l‐lysine monohydrochloride and l‐lysine sulfate produced using different strains of Corynebacterium glutamicum for all animal species based on a dossier submitted by FEFANA asbl
2019; Wiley; Volume: 17; Issue: 1 Linguagem: Inglês
10.2903/j.efsa.2019.5532
ISSN1831-4732
AutoresVasileios Bampidis, Giovanna Azimonti, Maria de Lourdes Bastos, Henrik Christensen, Birgit Dusemund, Maryline Kouba, Mojca Fašmon Durjava, Marta López‐Alonso, Secundino López Puente, Francesca Marcon, Baltasar Mayo, Alena Pechová, Mariana Petkova, Yolanda Sanz, Roberto Edoardo Villa, Ruud Woutersen, Lucio G. Costa, Noël Dierick, Gerhard Flachowsky, Boet Glandorf, Lieve Herman, Sirpa Kärenlampi, Ľubomír Leng, Alberto Mantovani, Robert John Wallace, Jaime Aguilera, Jordi Tarrés‐Call, Fernando Ramos,
Tópico(s)Genetically Modified Organisms Research
ResumoEFSA JournalVolume 17, Issue 1 e05532 Scientific OpinionOpen Access Safety of concentrated l-lysine (base), l-lysine monohydrochloride and l-lysine sulfate produced using different strains of Corynebacterium glutamicum for all animal species based on a dossier submitted by FEFANA asbl EFSA Panel on Additives and Products or Substances used in Animal Feed (EFSA FEEDAP Panel), EFSA Panel on Additives and Products or Substances used in Animal Feed (EFSA FEEDAP Panel)Search for more papers by this authorVasileios Bampidis, Vasileios BampidisSearch for more papers by this authorGiovanna Azimonti, Giovanna AzimontiSearch for more papers by this authorMaria de Lourdes Bastos, Maria de Lourdes BastosSearch for more papers by this authorHenrik Christensen, Henrik ChristensenSearch for more papers by this authorBirgit Dusemund, Birgit DusemundSearch for more papers by this authorMaryline Kouba, Maryline KoubaSearch for more papers by this authorMojca Kos Durjava, Mojca Kos DurjavaSearch for more papers by this authorMarta López-Alonso, Marta López-AlonsoSearch for more papers by this authorSecundino López Puente, Secundino López PuenteSearch for more papers by this authorFrancesca Marcon, Francesca MarconSearch for more papers by this authorBaltasar Mayo, Baltasar MayoSearch for more papers by this authorAlena Pechová, Alena PechováSearch for more papers by this authorMariana Petkova, Mariana PetkovaSearch for more papers by this authorYolanda Sanz, Yolanda SanzSearch for more papers by this authorRoberto Edoardo Villa, Roberto Edoardo VillaSearch for more papers by this authorRuud Woutersen, Ruud WoutersenSearch for more papers by this authorLucio Costa, Lucio CostaSearch for more papers by this authorNoël Dierick, Noël DierickSearch for more papers by this authorGerhard Flachowsky, Gerhard FlachowskySearch for more papers by this authorBoet Glandorf, Boet GlandorfSearch for more papers by this authorLieve Herman, Lieve HermanSearch for more papers by this authorSirpa Kärenlampi, Sirpa KärenlampiSearch for more papers by this authorLubomir Leng, Lubomir LengSearch for more papers by this authorAlberto Mantovani, Alberto MantovaniSearch for more papers by this authorRobert John Wallace, Robert John WallaceSearch for more papers by this authorJaime Aguilera, Jaime AguileraSearch for more papers by this authorJordi Tarrés-Call, Jordi Tarrés-CallSearch for more papers by this authorFernando Ramos, Fernando RamosSearch for more papers by this author EFSA Panel on Additives and Products or Substances used in Animal Feed (EFSA FEEDAP Panel), EFSA Panel on Additives and Products or Substances used in Animal Feed (EFSA FEEDAP Panel)Search for more papers by this authorVasileios Bampidis, Vasileios BampidisSearch for more papers by this authorGiovanna Azimonti, Giovanna AzimontiSearch for more papers by this authorMaria de Lourdes Bastos, Maria de Lourdes BastosSearch for more papers by this authorHenrik Christensen, Henrik ChristensenSearch for more papers by this authorBirgit Dusemund, Birgit DusemundSearch for more papers by this authorMaryline Kouba, Maryline KoubaSearch for more papers by this authorMojca Kos Durjava, Mojca Kos DurjavaSearch for more papers by this authorMarta López-Alonso, Marta López-AlonsoSearch for more papers by this authorSecundino López Puente, Secundino López PuenteSearch for more papers by this authorFrancesca Marcon, Francesca MarconSearch for more papers by this authorBaltasar Mayo, Baltasar MayoSearch for more papers by this authorAlena Pechová, Alena PechováSearch for more papers by this authorMariana Petkova, Mariana PetkovaSearch for more papers by this authorYolanda Sanz, Yolanda SanzSearch for more papers by this authorRoberto Edoardo Villa, Roberto Edoardo VillaSearch for more papers by this authorRuud Woutersen, Ruud WoutersenSearch for more papers by this authorLucio Costa, Lucio CostaSearch for more papers by this authorNoël Dierick, Noël DierickSearch for more papers by this authorGerhard Flachowsky, Gerhard FlachowskySearch for more papers by this authorBoet Glandorf, Boet GlandorfSearch for more papers by this authorLieve Herman, Lieve HermanSearch for more papers by this authorSirpa Kärenlampi, Sirpa KärenlampiSearch for more papers by this authorLubomir Leng, Lubomir LengSearch for more papers by this authorAlberto Mantovani, Alberto MantovaniSearch for more papers by this authorRobert John Wallace, Robert John WallaceSearch for more papers by this authorJaime Aguilera, Jaime AguileraSearch for more papers by this authorJordi Tarrés-Call, Jordi Tarrés-CallSearch for more papers by this authorFernando Ramos, Fernando RamosSearch for more papers by this author First published: 31 January 2019 https://doi.org/10.2903/j.efsa.2019.5532Citations: 5 Correspondence: feedap@efsa.europa.eu Requestor: European Commission Question number: EFSA-Q-2016-00391 Panel members: Giovanna Azimonti, Vasileios Bampidis, Maria de Lourdes Bastos, Henrik Christensen, Birgit Dusemund, Maryline Kouba, Mojca Kos Durjava, Marta López-Alonso, Secundino López Puente, Francesca Marcon, Baltasar Mayo, Alena Pechová, Mariana Petkova, Fernando Ramos, Yolanda Sanz, Roberto Edoardo Villa and Ruud Woutersen. Acknowledgements: The Panel wishes to thank the following for the support provided to this scientific output: Montserrat Anguita, Rosella Brozzi, Jaume Galobart, Matteo Innocenti, Paola Manini and Maria Vittoria Vettori. Legal notice: Relevant information or parts of this scientific output have been blackened in accordance with the European Commission decision on the confidentiality requests formulated by the applicant and further confidentiality requests formulated by the applicant for which a decision by the European Commission is pending. The blackened text will be subject to review once the full decision on the confidentiality requests is adopted by the European Commission. The full output was shared with the European Commission, EU Member States and the applicant. Adopted: 28 November 2018 AboutSectionsPDF ToolsExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat Abstract The EFSA FEEDAP Panel previously (2016) could not conclude on the safety of certain concentrated liquid l-lysine (base), l-lysine monohydrochloride (HCl) and l-lysine sulfate products manufactured using different strains of Corynebacterium glutamicum. New information on the safety of these products was provided by the applicant. The recipient strain C. glutamicum KCTC 12307BP qualifies for qualified presumption of safety (QPS) approach for safety assessment, the genetic modification does not introduce any safety concern and no introduced antibiotic resistance genes remain in its genome. Even if uncertainty remains concerning the absence/presence of the production strain and/or its recombinant DNA in the final products, these would not raise safety concerns. The liquid l-lysine (base) and l-lysine HCl produced by C. glutamicum KCTC 12307BP or C. glutamicum KCCM 11117P; and l-lysine HCl produced by C. glutamicum NRRL B-50547 are considered safe for the target species, consumers and the environment. Regarding the safety for the user, concentrated liquid l-lysine (base) and l-lysine HCl produced by C. glutamicum KCTC 12307BP, C. glutamicum NRRL B-50547 or C. glutamicum KCCM 11117P are not irritant to skin or eyes and they are not skin sensitisers. l-Lysine HCl is not hazardous by inhalation. The use of C. glutamicum DSM 24990 in the production of l-lysine sulfate is considered safe for the target species, consumers, users or the environment. No negative effects are to be expected for the target species within the proposed inclusion levels of 0.5–30 g lysine sulfate/kg complete feed provided that the total S intake complies with the recommendations of established scientific bodies. The use of C. glutamicum KCCM 10227 in the production of l-lysine sulfate is considered safe for the target species, consumers, users and the environment with regard to antimicrobial resistance. No negative effects are to be expected for the target species within common inclusion levels provided that the total S intake complies with the recommendations of established scientific bodies. 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 of concentrated liquid l-lysine (base), l-lysine monohydrochloride and l-lysine sulfate produced using different strains of C. glutamicum as a nutritional additive for all animal species based on the additional data submitted by the applicant. The approach followed by the FEEDAP Panel to assess the safety of concentrated liquid l-lysine (base), l-lysine monohydrochloride and l-lysine sulfate (solid or liquid) is in line with the principles laid down in Regulation (EC) No 429/2008 and the relevant guidance documents. Uncertainty remains concerning the absence/presence of the production strain C. glutamicum KCTC 12307BP and/or its recombinant DNA in the final products liquid l-lysine (base) and l-lysine HCl. The recipient strain qualifies for qualified presumption of safety (QPS) approach for safety assessment, the genetic modification does not introduce any safety concern and no introduced antibiotic resistance genes remain in the genome of the production strain. Therefore, the presence of viable cells and/or its recombinant DNA in the products would not raise safety concerns. The liquid l-lysine (base) and l-lysine HCl produced by C. glutamicum KCTC 12307BP are considered safe for the target species, consumers and the environment. The production strain C. glutamicum NRRL B-50547 carries a full gene of ■■■■■. Since no cells and recombinant DNA from the production strain were found in the product l-lysine HCl, the product does not raise safety concerns with respect to the genetic modification of the production strain provided that the downstream process is performed ensuring that no recombinant DNA of the production strain is present in the final product. l-Lysine HCl produced by C. glutamicum NRRL B-50547 does not raise safety concerns and it is considered safe for the target species, consumers and the environment. The products concentrated liquid l-lysine (base) and l-lysine monohydrochloride, produced using C. glutamicum KCCM 11117P, do not raise safety concerns with respect to the potential presence of viable cells of the production strain or its recombinant DNA. Consequently, the liquid l-lysine (base) and l-lysine HCl produced by C. glutamicum KCCM 11117P are considered safe for the target species, consumers and the environment. Regarding the safety for the user, concentrated liquid l-lysine (base) and l-lysine HCl produced by C. glutamicum KCTC 12307BP, C. glutamicum NRRL B-50547, C. glutamicum KCCM 11117P are not irritant to skin or eyes and they are not skin sensitisers. l-Lysine HCl is not hazardous by inhalation. The use of C. glutamicum DSM 24990 in the production of l-lysine sulfate is considered safe for the target species, consumers, users or the environment. No negative effects are to be expected for the target species within the proposed inclusion levels of 0.5–30 g lysine sulfate/kg complete feed provided that the total S intake complies with the recommendations of established scientific bodies. The use of C. glutamicum KCCM 10227 in the production of l-lysine sulfate is considered safe for the target species, consumers, users and the environment with regard to antimicrobial resistance. No negative effects are to be expected for the target species within common inclusion levels provided that the total S intake complies with the recommendations of established scientific bodies. 1 Introduction 1.1 Background and Terms of Reference as provided by the requestor Regulation (EC) No 1831/2003 establishes rules governing the Community authorisation of additives for use in animal nutrition and, in particular, Article 9 defines the terms of the authorisation by the Commission. The applicant, FEFANA asbl,1 is seeking a Community authorisation of concentrated liquid L-lysine (base), L-lysine monohydrochloride and L-lysine sulfate produced using different strains of Corynebacterium glutamicum to be used as a nutritional additive for all animal species. (Table 1). Table 1. Description of the substances Category of additive Nutritional additive Functional group of additive Amino acids, their salts and analogues Description Concentrated liquid L-lysine (base), L-lysine monohydrochloride and L-lysine sulfate produced using different strains of Corynebacterium glutamicum Target animal category All animal species Applicant FEFANA asbl Type of request New opinion On 02 December 2015, the Panel on Additives and Products or Substances used in Animal Feed of the European Food Safety Authority ("Authority"), in its opinion on the safety and efficacy of the product could not conclude on the safety of concentrated liquid L-lysine (base), L-lysine monohydrochloride and L-lysine sulfate produced using different strains of Corynebacterium glutamicum. The Commission has now received new data on the safety of concentrated liquid L-lysine (base), L-lysine monohydrochloride and L-lysine sulfate produced using different strains of C. glutamicum. In view of the above, the Commission asks the Authority to deliver a new opinion on the safety of concentrated liquid L-lysine (base), L-lysine monohydrochloride and L-lysine sulfate produced using different strains of C. glutamicum as a nutritional additive for all animal species based on the additional data submitted by the applicant. 1.2 Additional information The active substance of the three products under application, l-lysine, is produced either by genetically modified strains of C. glutamicum (KCTC 12307BP, NRRL B-50547 or KCCM 11117P) in the case of concentrated liquid l-lysine (base) and l-lysine monohydrochloride technically pure, or by chemically mutated strains of C. glutamicum (DSM 24990 or KCCM10227) in the case of l-lysine sulfate solid. The applicant withdrew the application for l-lysine sulfate (solid and liquid forms) produced by C. glutamicum DSM 16615 during the assessment. l-Lysine is currently authorised for its use in all animal species as a nutritional additive (functional group amino acids, their salts and analogues).2 No maximum content in feedingstuffs is established in the EU. The applicant has provided additional information on the characterisation of the additives, the characterisation of the production strains and on their safety. The initially proposed use in water for drinking of concentrated liquid l-lysine base and l-lysine monohydrochloride produced by C. glutamicum KCTC 12307BP was withdrawn by the applicant.3 Product from C. glutamicum rich in protein (inactivated fermentation by-product from the production of amino acids by culture of Corynebacterium glutamicum on substrates of vegetable or chemical origin, ammonia or mineral salts, it may be hydrolysed) with up to 0.3% antifoaming agents, 1.5% filtration/clarifying agents and 2.9% propionic acid and with a declared content of crude protein and propionic acid if > 0.5%; are listed in the Catalogue of feed materials (Commission Regulation (EU) 2017/1017).4 C. glutamicum is regarded as qualified presumption of safety (QPS) only when used as a production organism for amino acids (EFSA BIOHAZ Panel, 2013, 2017). 2 Data and methodologies 2.1 Data The present assessment is based on data submitted by the applicant in the form of additional information5 to a previous application of the same product.6 2.2 Methodologies The approach followed by the FEEDAP Panel to assess the safety of concentrated liquid l-lysine (base), l-lysine monohydrochloride and l-lysine sulfate (solid or liquid) is in line with the principles laid down in Regulation (EC) No 429/20087 and the relevant guidance documents: Guidance on nutritional additives (EFSA FEEDAP Panel, 2012a), Guidance for establishing the safety of additives for the consumer (EFSA FEEDAP Panel, 2012b), Guidance on studies concerning the safety of use of the additive for users/workers (EFSA FEEDAP Panel, 2012c), Guidance on the assessment of bacterial resistance to antibiotics of human or veterinary importance (EFSA FEEDAP Panel, 2012d), Guidance on the characterisation of microorganisms used as feed additives or as production organisms (EFSA FEEDAP Panel, 2018), Guidance on the risk assessment of genetically modified microorganisms and their products intended for food and feed use (EFSA GMO Panel, 2011) and Scientific Opinion on the maintenance of the list of QPS biological agents intentionally added to food and feed (EFSA BIOHAZ Panel, 2013, 2017 update). 3 Assessment The products under application are concentrated liquid l-lysine (base), l-lysine HCl, l-lysine sulfate solid and l-lysine sulfate liquid. l-Lysine is currently authorised for use in feeds for all animal species. The products under application are produced by fermentation with different C. glutamicum strains. They are intended to be used in all animal species as nutritional additives (functional group amino acids, their salts and analogues) in feed. l-Lysine HCl and the solid form of l-lysine sulfate are also proposed for their use in water for drinking. Concentrated liquid l-lysine base and l-lysine HCl are produced by microbial fermentation by genetically modified (GM) strains of C. glutamicum (KCTC 12307BP, NRRL B-50547 or KCCM 11117P). l-Lysine sulfate solid and liquid are produced by microbial fermentation by chemically mutated strains of C. glutamicum (DSM 24990 or KCCM10227). C. glutamicum is considered QPS approach for safety assessment for amino acid production when specific requirements are met (species identity confirmation and the absence of transmissible antibiotic resistance) (EFSA BIOHAZ Panel, 2013, 2017). When the production strain is a GM C. glutamicum, the genetic modification should not raise safety concerns. In the previous assessment, the FEEDAP Panel concluded that the use of l-lysine HCl technically pure produced using C. glutamicum NRRL B-50547 in animal nutrition is considered a hazard for the target species, consumer, user and environment due to the presence of a recombinant antimicrobial resistance gene in most batches of the product. The FEEDAP Panel could not conclude on: – The safety of the products concentrated liquid l-lysine (base) and l-lysine HCl, technically pure, produced using C. glutamicum KCCM 11117P (the presence of the production strain and/or its recombinant DNA in concentrated liquid l-lysine (base) and l-lysine HCl technically pure could not be excluded; and the genetic basis of the resistance of the production strain to clindamycin was not investigated) or KCTC 12307BP (incomplete data on its identity, safety of genetic modification, potential for antimicrobial resistance and the possible presence of the production strain and its recombinant DNA in the final products concentrated liquid l-lysine (base) and l-lysine HCl technically pure) for the target animal species, consumer, the user and the environment. – The safety of l-lysine sulfate solid and/or liquid originating from C. glutamicum KCCM 10227 or DSM 24990. The genetic base of resistance to at least one of the antimicrobials used in medical and veterinary practice was not sufficiently elucidated. The intrinsic high sulfate content in the product might have the potential to cause adverse effects in the target species. – The dermal sensitisation and on the irritancy of l-lysine HCl to skin or eyes. The concerns regarding the safety of the genetic modification and potential presence of transmissible resistance to antimicrobials might also have implications for the safety of the user. Regardless of the assessment of the genetic modifications, potential transmissible antimicrobial resistance or the absence of studies on safety for the user, the FEEDAP Panel had concerns on the safety of the administration of the amino acids, including l-lysine, via water for drinking for the target species because of the risk of imbalances and for hygiene reasons. Additional data have been submitted on the characterisation of the production strains and/or the products under assessment. These products had been characterised in a previous scientific opinion (EFSA FEEDAP Panel, 2016). The specifications of the products under assessment are: – Concentrated liquid l-lysine (base): minimum 50% lysine, maximum 48% water; – l-Lysine monohydrochloride: minimum 78% lysine, maximum 1.5% water; and – l-Lysine sulfate solid: minimum 40% lysine. 3.1 Concentrated liquid l-lysine (base) and l-lysine HCl produced by C. glutamicum KCTC 12307BP In its previous opinion (EFSA FEEDAP Panel. 2016), the FEEDAP Panel could not conclude on the safety of concentrated liquid l-lysine (base) and l-lysine HCl produced by the GM C. glutamicum KCTC 12307BP for the target animals, consumers and environment due to the lack of adequate information on the identity of the production strain, its genetic modification, its potential to harbour transferable antimicrobial resistance and lack of evidence of absence of the production strain or its recombinant DNA in the products. The applicant provided new information on the characterisation of the strain, the absence of the production strain or its recombinant DNA in the final product, characterisation of the products and studies on the safety of both products for the user. The new information provided is assessed below. 3.1.1 Characterisation of the C. glutamicum KCTC 12307BP and the absence of viable cells and DNA in the final products In the former assessment (EFSA FEEDAP Panel, 2016), evidence for the identity of the production strain was not submitted. In addition, no information on the process of genetic modification was provided. Antimicrobial susceptibility of the genetically modified microorganism C. glutamicum KCTC 12307BP was tested for antimicrobial susceptibility using ■■■■■ method. The battery of antimicrobials tested was that recommended by EFSA (EFSA FEEDAP Panel, 2012c). All minimum inhibitory concentration (MIC) values were below the corresponding cut-off for 'Other Gram +', except for clindamycin. The MIC is 2 mg/L and the cut-off 0.25 mg/L.8 No convincing explanation on the genetic basis of this resistance was provided. In the meantime, based in new scientific evidence, the FEEDAP Panel set the cut-off value of clindamycin from 0.25 to 4 mg/L for Corynebacterium, as reflected in the guidance on characterisation of microorganisms used as feed additives or as production organisms (EFSA FEEDAP Panel, 2018). Therefore, the strain is considered susceptible to all relevant antimicrobials, and of no concern for the target animals, users, consumers or the environment. Information on the parental/recipient organism The recipient organism was ■■■■■9 ■■■■■ The strain was identified by ■■■■■ analysis.10 Information regarding the donor organism All sequences introduced in the production strain derive from ■■■■■. Information regarding the genetic modification ■■■■■ ■■■■■ ■■■■■ ■■■■■ ■■■■■ ■■■■■ Absence of viable cells and DNA in the final product In the former assessment (EFSA FEEDAP Panel, 2016), no information on the absence of viable cells and DNA in the final products was provided. New data were provided in which the production strain was not found in three batches of each final product (l-lysine base and l-lysine HCl), by incubating in non-selective liquid medium at 30°C for 4 h, followed by plating on selective medium at 30°C for 10 days. However, the amount of sample analysed was not reported. Therefore, uncertainty remains on the absence of viable production cells in the final products.11 No recombinant DNA was found in three batches of each final product (l-lysine base and l-lysine_HCl), each tested in triplicate, by ■■■■■. However, no control DNA was added to the samples before DNA extraction as requested. Therefore, a limit of detection could not be established and uncertainty remains on the absence of recombinant DNA in the final products. 3.1.2 Characterisation of the additives New analytical data on impurities of concentrated liquid l-lysine (base) and l-lysine monohydrochloride (three batches each) have been submitted. All six batches yielded similar analytical results. As regards ■■■■■. Regarding the microbiological analyses, all batches were ■■■■■12 New certificates of analysis have been submitted showing aerobic plate count at 30°C < 102 CFU/g and the absence of vegetative anaerobic mesophilic bacteria in three batches of l-lysine HCl and concentrated liquid l-lysine base (50%).13 Regarding the physical properties, new data on particle size distribution (laser diffraction) of three batches of l-lysine HCl showed a fraction of particles < 10 μm ranging from 0 to 0.43% (v/v), a fraction of particles < 50 μm ranging from 0 to 2.2% (v/v) and a fraction of particles < 100 μm ranging from 0 to 5.3% (v/v).14 3.1.3 Safety of concentrated liquid l-lysine (base) and l-lysine monohydrochloride produced using C. glutamicum KCTC 12307BP Safety of the genetic modification Uncertainty remains concerning the absence/presence of the production strain and/or its recombinant DNA in the final products. Nevertheless, as the recipient strain C. glutamicum KCTC 12307BP qualifies for QPS approach for safety assessment, the genetic modification does not introduce any safety concern and no introduced antibiotic resistance genes remain in the genome of the production strain, the presence of viable cells and/or its recombinant DNA in the products do not raise safety concern. Safety for the target species, consumers and the environment The safety of l-lysine in animal nutrition when used in appropriate amounts is well established in the scientific literature. For nutritional additives produced by fermentation, the risk associated with the residues of the fermentation process in the final product needs to be assessed. The production strain C. glutamicum KCTC 12307BP has been unambiguously identified and it is susceptible to antimicrobials relevant for human or veterinary medicine. The recipient strain qualifies for QPS approach for safety assessment; the genetic modification does not introduce any safety concern and no introduced antibiotic resistance genes remain in the genome of the production strain. Although uncertainty remains concerning the possible presence of the production strain and/or its recombinant DNA in the final products, this does not raise safety concerns considering the above reasons. Consequently, the concentrated liquid l-lysine (base) and l-lysine HCl produced by C. glutamicum KCTC 12307BP are considered safe for the target species, consumers and the environment. Safety for the user In the former assessment (EFSA FEEDAP Panel, 2016), no specific studies concerning the user safety performed with either of the two products were available. Concentrated liquid l-lysine (base) was considered as corrosive due to its high pH. In the absence of data, the FEEDAP Panel could not conclude on the dermal sensitisation and on the irritancy of l-lysine HCl to skin or eyes. The applicant has provided new studies on the safety for the user performed with l-lysine HCl and concentrated liquid l-lysine produced by C. glutamicum KCTC 12307BP. Concentrated liquid l-lysine (base) Effects on skin and eyes An acute dermal irritation study was performed with concentrated liquid l-lysine (50%) in rabbits in accordance with OECD Guideline 404.15 None of the rabbits showed any skin reaction. Consequently, the product is classified as not irritating to human skin. An acute eye irritation study was conducted with concentrated liquid l-lysine (50%, pH 9.95) in rabbits, in accordance with OECD Guideline 405.16 Some redness of conjunctiva and chemosis was observed within 24 h post-application in different grades in all three rabbits. As the recorded scores were lower than the threshold value for classifying a product as an eye irritant (0.7 vs 2), the test material is considered not irritant for human eye. The skin sensitisation potential of the product concentrated liquid l-lysine (50%) was studied in guinea pigs in accordance with OECD Guideline 406 (Guinea pig maximisation test).17 No dermal reactions were observed at the 24- or 48-h post-challenge. Consequently, the test item has no sensitising properties. l-lysine monohydrochloride Effects on the respiratory system Although the additive has very low fraction of particles below 100 μm (up to 5.3%), its dusting potential is high (7.1 g/m3, EFSA FEEDAP Panel, 2016), indicating that exposure of the user by inhalation is possible. An acute inhalation toxicity study in accordance with OECD Guideline 436 was performed with l-lysine HCl (99.4% pure).18 Six CRL (WI) BR rats (3 males and 3 females) were exposed by inhalation ('nose only') to a concentration of 5.1 mg l-lysine HCl/L air for 4 h. No mortality was observed. Dyspnoea during or shortly after exposure (all rats, 4 of them up to day 1) or weak grip and limb tone (one female rat, the first days of the observation period) were observed. No signs were observed from day 6 to the end of the study. Only one rat showed a slight body weight loss (2 g) during da
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