This guidance document is intended to assist the applicant in the preparation and the presentation of an application, as foreseen in Article 7.6 of Regulation (EC) No 1831/2003, for the authorisation of additives used in animal nutrition. It specifically covers the assessment of the safety for the environment.

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Little is known about the environmental hazards linked to the treatment of farm animals with antibiotics and subsequent spreading of manure, especially regarding soil microbial communities. In this investigation, pollutioninduced community tolerance (PICT) of bacteria from soils artificially spiked with the sulfonamide sulfachloropyridazine (SCP) was investigated. Tolerance of the bacterial communities after 3 weeks' exposure to SCP was determined by analyzing the sensitivity of 31 microbial metabolic processes in microtiter plates. Bacterial suspensions extracted from soils containing higher concentrations of SCP showed an increased tolerance of their metabolic activities to this antibiotic. An increase in tolerance by 10% was found at 7.3 mg/kg dw SCP. The PICT effect could be demonstrated by both a shift in the tolerance of the average of all metabolic activities and a shift of the physiological process sensitivity distributions made up from the single metabolic processes. The PICT effect was accompanied by smaller changes in the communitylevel physiological profile (CLPP). To conclude, PICT has been shown to be a versatile and illustrative method for the detection of the effects of antibacterial agents on soil microorganisms.

ABSTRACT. We present an overview of the risks that underground thermal energy storage (UTES) can impose on the groundwater system, drinking water production, and the subsurface environment in general. We describe existing policy and licensing arrangements for UTES in the Netherlands, as well as the capability of the current and future Dutch policy and legal framework to minimize or mitigate risks from UTES on groundwater resources. A survey at the European Union member state level indicates that regulation and research on the potential impacts of UTES on groundwater resources and the subsurface environment often lag behind the technological development of and ever-growing demand for this renewable energy source. The lack of a clear and scientifically underpinned risk management strategy implies that potentially unwanted risks might be taken at vulnerable locations such as near well fields used for drinking water production, whereas at other sites, the application of UTES is avoided without proper reasons. This means that the sustainability of UTES as a form of renewable energy is currently not fully understood, and the technology may be compromising the natural resilience of the subsurface environment. We recognize three main issues that should be addressed to secure sustainable application of UTES: Scientific research is required to further elucidate the impacts of UTES on groundwater; cross-sectoral subsurface planning is required to minimize negative conflicts between UTES and other subsurface interests; and EU-wide guidelines and standards are required for quality assurance and control when installing UTES systems.

Lysine is an essential amino acid for all animal species; it is the first limiting amino acid in swine nutrition and the second limiting amino acid in poultry nutrition. L-Lysine and its salts are widely used in the feed industry to optimise dietary protein.Neither the production strain nor its recombinant DNA was detected in any of the final products. The final products do not raise any safety concern with regard to the genetic modifications.Concentrated liquid L-lysine (base), concentrated liquid L-lysine HCl and L-lysine HCl technically pure are considered safe for target species when supplemented in appropriate amounts.Lysine produced by E. coli (FERM BP-10941) is not genotoxic and the results of subchronic studies do not indicate any specific concerns. As there are no lysine metabolites associated with safety concerns in the animal tissues and products, the FEEDAP Panel considers that the use of L-lysine and its hydrochloride salts in animal feed does not pose a risk for the consumer.Concentrated liquid L-lysine (base), concentrated liquid L-lysine HCl and L-lysine HCl technically pure are not considered to have the potential to cause respiratory toxicity, skin or eye irritation or skin sensitisation, but respiratory sensitisation cannot be excluded.L-Lysine is a substance naturally occurring in bacteria, plants and animals. The use of L-lysinecontaining feed additives does not represent a risk to the environment.Concentrated liquid L-lysine (base), concentrated liquid L-lysine HCl and L-lysine HCl technically pure are considered equivalent in terms of L-lysine availability to the target animals. The efficacy of supplementing L-lysine and its hydrochloride salts is extensively demonstrated in the literature for mammals (except ruminants), poultry and fish, including its use in a liquid or a powder form. Therefore, it does not require any further demonstration. Response in ruminants requires some degree of protection of L-lysine from ruminal degradation.

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Methanopterin is a coenzyme involved in methanogenesis. From 2 kg wet cells of Methanobacterium thermoautotrophicum about 35 μmol methanopterin were isolated. The structure of this compound was elucidated by various two‐dimensional nuclear‐magnetic‐resonance techniques. Methanopterin was identified as N‐[1′‐(2″‐amino‐4″‐hydroxy‐7″‐ methyl‐6″‐ pteridinyl)ethyl]‐4‐[2′,3′,4′,5′‐ tetrahydroxypent‐1′‐ yl (5′→1″) O‐α‐ribofuranosyl‐5″‐phosphoric acid] aniline, in which the phosphate group is esterified with α‐hydroxyglutaric acid. The molecular formula of the sodium salt of methanopterin at pH 7.0 is C30H38O16N6PNa3·xH2O (x is about 4). The anhydrous sodium salt of methanopterin has a molecular mass of 838.60 Da and the molar absorption coefficient at 342 nm is 7.4 mM−1 cm−1 at pH 7.0.