1987L0153 — EN — 26.10.2001 — 003.001 — 1
This document is meant purely as a documentation tool and the institutions do not assume any liability for its contents
►B COUNCILDIRECTIVE
of16February1987
fixingguidelinesfortheassessmentofadditivesinanimalnutrition
(87/153/EEC)
(OJ L 64, 7.3.1987, p. 19)
Amended by:
Official Journal
No page date
►M1CommissionDirective 94/40/EC of 22 July 1994 L 208 15 11.8.1994
►M2CommissionDirective 95/11/EC of 4 May 1995 L 106 23 11.5.1995
►M3CommissionDirective 2001/79/EC of 17 September 2001 L 267 1 6.10.2001
▼B
COUNCILDIRECTIVE
of16February1987
fixingguidelinesfortheassessmentofadditivesinanimalnutrition
(87/153/EEC)
THE COUNCIL OF THE EUROPEAN COMMUNITIES,
Having regard to the Treaty establishing the European Economic
Community,
Having regard to Council Directive 70/524/EEC of 23 November 1970
concerning additives in feedingstuffs(1), as last amended by Commis-
sionDirective 86/525/EEC(2), and in particular Article 9 thereof,
Having regard to the proposal from the Commission,
Whereas Directive 70/524/EEC provides that the examination of addi-
tives must be performed onthe basis of a dossier forwarded officially
to the Member States and to the Commission;
Whereas such dossiers must make it possible to verify that additives
comply, in respect of their proposed use, with the general principles
laid down in the Directive for their inclusion in the Annexes thereto;
Whereas it has been found necessary to provide for the dossiers to be
compiled in accordance with common guidelines defining the scientificdata which make it possible to identify and characterize the productsconcerned and the studies necessary in order to evaluate, in particular,their efficacy and their safety for man, animals and the environment;
Whereas the guidelines are intended primarily as a general guide;
whereas, depending on the nature of the additive or its conditions ofuse, the extent of the studies necessary in order to evaluate its proper-ties or its effects may vary;
Whereas it is indispensable to apply the principles of good laboratory
practice when developing additives intended for use in feedingstuffs to
ensure that the results of laboratory tests are not disputed; whereas
recourse to procedures involving the use of laboratory animals forexperimental or other scientific purposes should be kept to a minimum;
Whereas the guidelines have been drawn up on the basis of present
scientific and technical knowledge and they may be adapted if neces-sary to any developments in this sphere,
HAS ADOPTED THIS DIRECTIVE:
Article 1
Member States shall prescribe that the dossiers which must accompany
every request for the inclusion of an additive or a new use of an addi-tive in the Annexes to Directive 70/524/EEC are to be compiled in
accordance with the guidelines set out in the Annex to this Directive.
Article 2
This Directive shall apply without prejudice to provisions on:
(a) good laboratory practice for the purposes of mutual acceptance of
data for the evaluationof chemical products; an d
(b) the protection of animals used for experimental or other scientific
purposes.1987L0153 — EN — 26.10.2001 — 003.001 — 2
(1) OJ No L 270, 14. 12. 1970, p. 1.
(2) OJ No L 310, 5. 11. 1986, p. 19.
▼B
Article 3
Member States shall bring into force the laws, regulations or adminis-
trative provisions necessary in order to comply with this Directive by
31 December 1987 at the latest. They shall forthwith inform the
Commissionthereof.
Article 4
This Directive is addressed to the Member States.1987L0153 — EN — 26.10.2001 — 003.001 — 3
▼M1
ANNEX
GUIDELINES FOR THE ASSESSMENT OF ADDITIVES IN FEEDING-
STUFFS
PART I
ADDITIVESOTHERTHANMICRO-ORGANISMSANDENZYMES
GENERAL ASPECTS
This document is intended as a guideline for establishing dossiers on substances
and preparations being submitted for authorisation as additives in feedingstuffs
or a new usage of an authorised additive. The term ‘additive’, as used in these
guidelines refers to the active chemically specified substances or the prepara-
tions containing active substances in the state in which they will be
incorporated in premixtures and feedingstuffs. The dossiers must enable an
assessment to be made of the additives based on the present state of knowledge
and make it possible to ensure their compliance with the fundamental principles
laid down for their authorisation, which are the subject of the provisions of
Article 3a of Council Directive 70/524/EEC.
Where a dossier concerns an additive consisting of or containing genetically
modified organisms within the meaning of Articles 2(1) and (2) of Council
Directive 2001/18/EC(1), the dossier must include the additional information
specified inArticle 7a(1) of Directive 70/524/EEC, inadditionto the in forma-
tionrequired by these guidelin es.
The dossiers should include detailed reports of all studies done, presented in the
order and with the numbering proposed in these guidelines. They should include
references and copies of all published scientific data relevant to the evaluation
of the additive. Anelectron ic versionof the dossier should be made available.
The studies are intended to demonstrate the safety of use of the additive in rela-
tionto:
(a) the target species at the proposed levels of incorporation in the feedingstuff;
(b) those likely to be exposed to the additive by respiratory, other mucosal, eye
or cutaneous contact while handling the additive as such or incorporated
into premixtures or feedingstuffs;
(c) consumers who ingest food products obtained from animals having received
the additive, which could contain residues of the additive, or its metabolites;
this will generally be ensured by the setting of maximum residue limits
(MRLs) and withdrawal periods;
(d) the animals and the human-beings through the selection and spread of anti-
microbial resistance genes;
(e) the environment arising from the additive itself or by products derived from
the additive, either directly and/or excreted by animals.
As a general rule, studies to establish the identity, conditions of use, physico-
chemical properties, methods of determination and efficacy of the additive, and
also its metabolic fate and residues, physiological and toxicological effects on
target species must be provided. When the additive is intended for a category
of animals belonging to a defined species, efficacy and residue studies must be
performed onthis target category. The studies n ecessary for the evaluationof
risks to human health or the environment will depend essentially on the nature
of the additive and the circumstances of its use. In this respect, no strict rule is
applicable. If necessary, additional information will be requested. Reasons must
be givenfor the omissionfrom the dossier of an y data prescribed inthese guide-
lines. In particular, studies of mutagenicity, carcinogenicity and reproductiontoxicity studies may only be dispensed with if the chemical composition, prac-
tical experience, or other considerations can reasonably exclude these effects.
The studies should be done and reported according to appropriate quality stan-
dards (e.g. good laboratory practice (GLP) pursuant to Council Directive 87/18/
EEC of 18 December 1986, on the harmonisation of laws, regulations and
administrative provisions relating to the application of the principles of good
laboratory practice and the verification of their applications for tests on
chemical substances(
2)).
Expert reports onquality, efficacy an d safety should be provided. Their authors,
who should have relevant qualifications and be recognised experts in the field
concerned, should not have been personally involved in the conduct of the tests1987L0153 — EN — 26.10.2001 — 003.001 — 4
(1) OJ L 106, 17.4.2001, p. 1.
(2) OJ L 15, 17.1.1987, p. 29.▼M3
▼M3
included in the dossier. The reports must provide a critical appraisal of the
documentation provided by the applicant; a factual summary is not sufficient.
The determination of physico-chemical, toxicological and eco-toxicological
properties shall be performed inaccordan ce with the methods established by
Council Directive 67/548/EEC of 27 June 1967 on the approximation of laws,
regulations and administrative provisions relating to the classification, packaging
and labelling of dangerous substances(1), as last amended by Commission
Directive 2000/33/EC(2), or with updated methods recognised by international
scientific bodies. The use of methods other than these must be justified.
Each dossier shall contain an adequate summary, an annex proposal and may
contain a monograph. The dossiers relating to antibiotics, coccidiostats and other
medicinal substances and to growth promoters must be accompanied by a mono-
graph, conforming to the model provided in Section V, enabling the additive
concerned to be identified and characterised in accordance with Article 9n of
Directive 70/524/EEC. An identification note conforming to the model in
SectionVI has to be provided for all additives.
For additives intended exclusively for pet food it may not always be necessary
to subject additives to an as exhaustive program of chronic toxicity, mutageni-
city, reproductive toxicity and carcinogenicity testing as that required for
additives intended for feeding to livestock from which products for human
consumption are derived. Residue studies in pet animals are not required.
Study of the metabolic fate of the additive in food producing target animals and
inlaboratory species used for toxicity testin g, is required inorder to:
(a) ensure that there is adequate data on the toxicity of the parent additive and
any metabolites produced in the target species to which the consumer might
be exposed. To this end a comparison of the metabolic fates of the additive
in the target and laboratory animal species used for the toxicity testing is
important;
(b) identify and quantify the appropriate marker residue(s) to be used for setting
the MRL for the marker residue and the withdrawal periods for the final
product.1987L0153 — EN — 26.10.2001 — 003.001 — 5
(1) OJ L 196, 16.8.1967, p. 1.
(2) OJ L 136, 8.6.2000, p. 90.
▼M3
CONTENTS
1.SectionI:Summaryofthedatainthedossier
2.Section II: Identity, characterisation and conditions of use of the
additive;methodsofcontrol
2.1. Identity of the additive
2.2. Characterisationof the active substan ce(s)2.3. Characterisation of the additive: Physico-chemical and technological
properties
2.4. Conditions of use of the additive2.5. Control methods
3.SectionIII:Studiesconcerningtheefficacyoftheadditive
3.1. Studies onthe effects onfeedin gstuffs
3.2. Studies onthe effects onan imals3.3. Studies onthe quality of an imal produce
3.4. Studies onthe effects onthe characteristics of an imal wastes.
4.SectionIV:Studiesconcerningthesafetyofuseoftheadditive
4.1. Studies ontarget species
4.2. Studies onlaboratory an imals
4.3. Safety evaluationfor the humancon sumer4.4. Worker safety assessment
4.5. Environmental risk assessment
5.SectionV:Formofmonograph
5.1. Identity of the additive5.2. Specifications concerning the active substance
5.3. Physico-chemical, technological and biological properties of the addi-
tive.
5.4. Control methods
5.5. Biological properties of the additive5.6. Details of the quantitative and qualitative residues in target tissues, if
any, found in animal produce following envisaged use of the additive.
5.7. The ADI, the established MRLs and the withdrawal period should be
given, if appropriate.
5.8. Other characteristics suitable for identification of the additive5.9. Conditions of use
5.10. Date
6.SectionVI:Formofidentification note
7.Section VII: Renewal of authorisation of additives submitted to a
brand-specificapproval
8.SectionVIII:Newapplicantrelying onthefirstauthorisationofan
additive whose authorisation is linked to a person responsible for
puttingthemintocirculation1987L0153 — EN — 26.10.2001 — 003.001 — 6
▼M3
1.SectionI:Summaryofthedatainthedossier
The summary must follow the order of the guidelines and address all
the different parts with reference to the relevant pages of the dossier.
It should contain a proposal covering all the conditions for the author-
isationsought.
2.Section II: Identity, characterisation and conditions of use of the
additive;methodsofcontrol
2.1. Identity of the additive
2.1.1. Proposed proprietary name(s)
2.1.2. Type of additive according to its main function. When possible,
evidence of mode(s) of action should be included. Any other uses of
the active substance should be specified.
2.1.3. Qualitative and quantitative composition (active substance, other
components, impurities, batch-to-batch variation). If the active
substance is a mixture of active components, each of which is clearly
definable, the main components must be described separately and the
proportions in the mixture given.
2.1.4. Physical state, particle size distribution, particle shape, density, bulk
density; for liquids: viscosity, surface tension.
2.1.5. Manufacturing process including any specific processing procedures.
2.2. Characterisation of the active substance(s)
2.2.1. Generic name, chemical name according to IUPAC nomenclature
(International Union of Pure and Applied Chemistry), other generic
international names and abbreviations. Chemical abstracts service
number (CAS).
2.2.2. Structural formula, molecular formula and molecular weight.
For active substances being fermentation products: microbial origin
(name and place of culture collection recognised as an international
depository authority, preferably in the European Union, where the strain
is deposited, accession number and all relevant morphological, physio-
logical, genetic and molecular characteristics for its identification). For
genetically modified strains, information on the genetic modification
must be provided.
2.2.3. Purity
Identification and quantification of occurring chemical and microbial
impurities and toxic substances, confirmation of the absence of produc-
tionorgan isms.
2.2.4. Relevant properties
Physical properties of the chemically specified substances: dissociation
constant, pKa, electrostatic properties, melting point, boiling point,
density, vapour pressure, solubility in water and organic solvents, Kowand Koc, mass and absorption spectra, NMR data, possible isomers and
any other appropriate physical properties.
2.2.5. Manufacturing, purification processes, media used and, for fermentation
products, batch-to-batch variation.
2.3. Characterisation of the additive: Physico-chemical and technological
properties
2.3.1. Stability of each formulation of the additive on exposure to environ-
mental conditions such as light, temperature, pH, moisture, oxygen and
packing material. Expected shelf life of the additive as marketed.
2.3.2. Stability of each formulation of the additive during the preparation and
storage of premixtures and feedingstuffs, in particular stability to antici-
pated process/storage conditions (heat, moisture, pressure/shear, time
and packing material). Possible degradation or decomposition products.
Expected shelf life of the additive.
2.3.3. Other appropriate physico-chemical or technological properties in order
to obtain and keep homogeneous mixtures in premixtures and feeding-
stuffs, antidusting and electrostatic properties, dispersability in liquids.
2.3.4. Incompatibilities or interactions that could be expected with feeding-
stuffs, carriers, other approved additives or with medicinal products.1987L0153 — EN — 26.10.2001 — 003.001 — 7
▼M3
2.4. Conditions of use of the additive
2.4.1. Where an additive has significant technological as well as zootechnical
effects it has to meet the requirements of both claims. The claims for
each additive have to be identified and justified.
2.4.2. Proposed technological use in animal feedingstuff manufacture or if
appropriate inraw materials.
2.4.3. Proposed mode of use in animal nutrition (e.g. animal species or cate-
gories and age group/production stage of animal, type of feedingstuff,
and contra-indications).
2.4.4. Proposed method and level of inclusion in premixtures and feeding-
stuffs or raw materials if appropriate expressed as proportionof the
additive and chemically specified substances by weight for premixtures,
for feedingstuffs or raw materials if appropriate, with proposed dose in
the final feedingstuff and proposed duration of administration and with-
drawal period if appropriate.
2.4.5. Data from other known uses of the active substance (e.g. in foodstuffs,
human or veterinary medicine, agriculture and industry) must be
provided.
2.4.6. Proposed material safety data sheet as foreseenby CommissionDirec-
tive 91/155/EEC(1) defining and laying down the detailed arrangements
for the system of specific information relating to dangerous preparations
inimplemen tationof Article 10 of Directive 88/379/EEC(2) and, if
necessary, proposed measures for the prevention of occupational risks
and means of protection during manufacture, handling, use and
disposal.
2.5. Control methods
2.5.1. Descriptionof the methods used for the determin ationof the criteria
listed under items 2.1.3, 2.1.4, 2.2.3, 2.2.4, 2.3.1, 2.3.2, 2.3.3 and 2.3.4.
2.5.2. Description of the qualitative and quantitative analytical methods for
routine control of the active substance in premixtures and feedingstuffs.
This method has to be validated in a ring test involving at least four
laboratories or has to be validated in house following international
harmonised guidelines for the in-house validation of methods of
analysis(3) with respect to the following parameters: applicability,
selectivity, calibration, accuracy, precision, range, limit of detection,
limit of quantification sensitivity, robustness and practicability.
Evidence that these characteristics have been assessed must be made
available (2.5.4).
2.5.3. Description of the qualitative and quantitative analytical methods for
determining the marker residue(s)(4) of the active substance in target
tissues and animal products.
2.5.4. The methods mentioned under 2.5.2 and 2.5.3 should be accompanied
by information as to the sampling method used, percentage recovery,
specificity, accuracy, precision, limits of detection, limits of quantifica-
tion and validation procedure used. Reference standards of the active
substance and/or of the marker residue(s) must be available as well as
information on the optimum storage conditions for these reference stan-
dards. When devising methods, consideration must be given to the fact
that their limits of quantification must be below the MRLs. Moreover,
their suitability for routine analysis must be taken into account.
3.SectionIII:Studiesconcerningtheefficacyoftheadditive
3.1. Studies on the effects on feedingstuffs
These studies concern technological additives such as antioxidants,
preservatives, binders, emulsifiers, stabilisers, gelling agents, pH modi-
fiers etc., which are intended to improve or stabilise the characteristics
of premixtures and feedingstuffs but have no direct biological effect on
animal production. All claimed activities or effects of the additive have
to be justified by scientific information.1987L0153 — EN — 26.10.2001 — 003.001 — 8
(1) OJ L 76, 22.3.1991, p. 35.
(2) OJ L 187, 16.7.1988, p. 14.
(3) Method Validation— A Laboratory Guide, Eurachem Secretariat, Laboratory of the
Government Chemist, Teddington, United Kingdom, 1996.
(4) The marker residue is a residue whose concentration is in known relationship with the
rate at which the concentration of the total residue in the target tissue depletes to the
MRL.
▼M3
Evidence of the efficacy of the additive must be provided by means of
appropriate criteria as reflected in recognised acceptable methods, under
the intended conditions of use in comparison with appropriate control
feedingstuffs. These investigations must be designed and performed so
as to permit a statistical evaluation.
Full information on the active substances, preparations, premixes and
feedingstuffs examined, the reference number of the batches, the
detailed treatment and testing conditions should be provided. Positive
and negative effects, both technological and biological, should be
described for each experiment.
3.2. Studies on the effects on animals
Studies onzootechn ical additives must be performed intarget species/
animal categories for which the additive is intended in comparison with
negative control groups (without antibiotics, growth promoters or other
medicinal substances) and, possibly, with groups receiving feedingstuffs
containing EU approved additives of known effectiveness used at their
recommended dosages (positive control).
Animals used should be healthy and preferably from a homogeneous
group.
Studies must permit the evaluationof the efficacy of the additive
according to farming practice in the EU. Similar protocol designs
should, where possible, be used for all trials so that data caneven tually
be tested for homogeneity and pooled (if tests so indicate) for statistical
evaluation.
No single design is recommended, flexibility being provided to allow
for scientific discretion in the design and conduct of the studies. The
experimental design used must be justified according to the claim for
the use of the additive and must include consideration of adequate
statistical power.
3.2.1. For coccidiostats and other medicinal substances
Importance should primarily be attached to evidence of the specific
effects (e.g. species controlled, life-cycle stage(s) affected) and particu-
larly to prophylactic properties (e.g. morbidity, mortality, oocyst count
and lesion score).
Information on the effect on feed efficiency and liveweight gain should
be provided.
The required efficacy data involve three stages of target animal experi-
mentation:
(a) controlled battery-cage experiments (single and mixed infections);
(b) controlled floor pen studies (simulated use conditions);
(c) controlled field trials (actual use conditions).
Simultaneously and when relevant, within trials on efficacy, additional
data should be recorded to allow an evaluation of interference with
growth and feed conversion (fattening birds, replacement layers and
rabbits), effects on egg fertility and hatchability (breeding birds).
3.2.2. For other zootechnical additives
In formationshould be provided onthe effects onfeed in take, body
weight, feed efficiency (preferably on a dry matter basis), product
quality and yield and any other parameter of benefit to the animal, the
environment, the producer or the consumer. The studies should include
an indication of dose/response relationship where appropriate.
3.2.3. Experimental conditions
Trials should be carried out at least at two different locations. They
should be reported individually, giving details of the controls and each
experimental treatment. The trial protocol should be carefully drawn up
with regard to general descriptive data as follows:
3.2.3.1. Herd or flock: location and size; feeding and rearing conditions, method
of feeding; for aquatic species, size and number of tanks or pens at the
farm and water quality.
3.2.3.2. Animals: species (for aquatic species intended for human consumption
identification shall be made by their colloquial name followed in
parenthesis by the latin or Linnean description), breed, age, sex, identi-
fication procedure, physiological stage and general health.1987L0153 — EN — 26.10.2001 — 003.001 — 9
▼M3
3.2.3.3. Number of test and control groups, number of animals in each group.
The number of animals involved in the trials must permit statistical
analysis. The methods of statistical evaluation used should be stated.
At least three independent comparable trials at the level of p < 0,05 in
each of the claimed animal category(ies) have to be provided to show
the effect referred to. In the case of ruminants a lower level of prob-
ability could be accepted p < 0,10. The report should include all
animals or experimental units involved in the trials. Cases which cannot
be assessed due to a lack or loss of data should be reported and their
distributionwithinthe groups of an imals classified.
3.2.3.4. Diets: description of manufacture and quantitative composition of the
diet(s) in terms of ingredients used, relevant nutrients (analysed values)
and energy. Feed intake records.
3.2.3.5. Concentration of the active substance (and, where that is the case,
substances used for comparative purposes) in the feedingstuffs should
be established by a control analysis, using the appropriate recognised
method. Reference number(s) of the batches.
3.2.3.6. Date and exact duration of testing. Date and nature of the examinations
performed.
3.2.3.7. Dose determination studies: the purpose of these studies is to explain
the rationale for the selection of a dose or dose range claimed to be
optimally effective. Dose determination will be based on a control
(without antibiotics, growth promoter or other medicinal substances)
and at least three non-zero levels in target animals.
3.2.3.8. The timing and prevalence of any undesirable consequences of treat-
ment in individuals or groups must be reported (give details of the
observationprogramme used inthe study).
3.2.3.9. All additives studied under farm conditions must have good scientific
evidence of safety for the user, consumer, animal and the environment.
Where an additive does not meet the requirements for consumer safety
any study undertaken should be designed to prevent animal products
derived from the test animals from entering the human food chain.
3.3. Studies on the quality of animal produce
Animal products should be examined for organoleptic, nutritional,
hygienic and technological qualities as appropriate.
3.4. Studies on the effects on the characteristics of animal wastes
If the additive is intended to modify some characteristics of animal
waste (e.g. nitrogen, phosphorus, odour, volume), then studies demon-
strating these properties are required.
4.SectionIV:Studiesconcerningthesafetyofuseoftheadditive
The studies outlined in this section are intended to permit assessment
of:
— the safety of use of the additive inthe target species,
— any risk associated with the selection and/or transfer of resistance to
antibiotics and increased persistence and shedding of enteropatho-
gens,
— the risks to the consumer which could result from the consumption
of food containing residues of the additive or its metabolites,
— the risks from respiratory, other mucosal, eye or cutaneous contact
for persons likely to handle the additive as such or as incorporated
into premixtures or feedingstuffs,
— the risks of adverse effects on the environment, from the additive
itself or by products derived from the additive, either directly and/
or excreted by animals.
Consideration should be given to known incompatibilities and/or inter-
actions between the additive and veterinary medicines and/or
components of the diet relevant to the species concerned.
These studies will normally be required in their entirety for each addi-
tive unless a specific exclusion or modification is specified in the
Directive.
A more limited submissionwill n ormally be accepted for a proposed
extension of the authorised use to a species which is physiologically
and metabolically close to one in which the use of the additive has
already beenapproved. This reduced data set should demon strate safety
to the new species and lack of significant differences in metabolic fate1987L0153 — EN — 26.10.2001 — 003.001 — 10
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and residues in edible tissues. The proposed MRL and withdrawal
period for the species must be justified.
In order to assess the risks for the consumer and consequently the deter-
mination of the MRLs and the withdrawal period the following
information has to be provided:
— the chemical structure of the active substance,
— the metabolism inthe proposed target species,— the nature of the residues in these target species,— tissue depletionstudy of residues,— data onthe biological effects of the active substan ce together with
its metabolites.
A knowledge of the bioavailability of the residues (both unbound and
bound) may also be useful namely when many metabolites are
produced and no marker residues is evidenced (see section 4.1.3.3).
Furthermore, knowledge of the composition and of the physico-
chemical and biological properties of the major excreted materials
deriving from the additive are required to define the extent of the
studies necessary for assessment of the risk of adverse effects on the
environment or persistence in the environment (see paragraph 4.5).
4.1. Studies on target species
4.1.1. Tolerance tests on target species/animal categories
The aim is to determin e a safety margin(i.e. marginbetweenthe
maximum proposed dose-level in feedingstuffs and the minimum level
resulting in unfavourable effects). However a safety margin of a factor
of at least 10 is considered sufficient to require no further testing. Such
a tolerance test must be conducted in the target species/animal cate-
gories preferably through the entire length of the production period
although a test period of one month would normally be acceptable.
This requires at least the assessment of clinical signs and other para-
meters to ascertain effects on target animal health. A negative control
group (without antibiotics, growth promoter or other medicinal
substances) has to be included. Depending on the toxicological profile
additional parameters may also be required. Any adverse effects
detected during efficacy trials should also be reported in this section.
Whenever the product is intended for use in animals which may be used
for breeding, studies should be conducted to identify possible impair-
ment of male or female general reproductive function or harmful
effects on progeny resulting from the administration of the additive
under investigation.
4.1.2. Microbiological safety of the additive.4.1.2.1. All studies should be performed with the highest proposed dose level
4.1.2.2. If the active substance possesses antimicrobial activity at feed concen-
tration level, the minimum inhibitory concentration (MIC) should be
determined in appropriate pathogenic and non-pathogenic, endogenous
and exogenous bacteria, according to standardised procedures.
4.1.2.3. Tests to determine the ability of the additive to:
— induce cross-resistance to relevant antibiotics,
— select resistant bacterial strains under field conditions in the target
species and, if so, investigations on the genetic mechanisms for
transfer of the resistance genes.
4.1.2.4. Tests to determine the effect of the additive:
— on a number of opportunistic pathogens present in the digestive
tract (e.g. enterobacteriaceae ,enterococci andclostridia ),
— on the shedding or excretion of relevant zoonotic micro-organisms,
e.g.salmonella spp,campylobacter spp.
4.1.2.5. In cases where the active substance shows an antimicrobial action, field
studies to monitor for bacterial resistance to the additive should be
provided.
4.1.3. Metabolism and residue studies
4.1.3.1. The aim of the studies is to:
— establish the metabolic pathways of the active substance as a base
for its toxicological evaluation,
— identify residues and establish their kinetics in the edible tissues and
products (milk, eggs),1987L0153 — EN — 26.10.2001 — 003.001 — 11
▼M3
— identify the excreted substances as a prerequisite for assessing their
impact on the environment.
Occasionally, e.g. for fermentation-derived additives, it could be neces-
sary to extend these studies to other substances added to or derived
during the fermentation process. Possession of a toxicity significant in
relation to that of the active component(s) of the additive would exem-
plify this circumstance.
4.1.3.2. Pharmacokinetics
The planning and experimental design of the studies must take into
account the anatomical, physiological (age, type, sex), zootechnical
category and environmental peculiarities of the target population.
When appropriate, the influence of the intestinal or ruminal microflora,
enterohepatic circulation or caecotrophy must be considered. The dose
regimen tested must be that intended for use, and possibly a multiple of
that dose if justified. The active substance (including the labelled
substance) must be incorporated into the feed unless there is justifica-
tion for not doing so.
The studies required are the following:
— metabolic balance and kinetics in the plasma/blood following a
single dose administration in order to assess the rate and extent of
the absorption, distribution and excretion (urine, faeces, gills, bile,
expired air, milk or eggs),
— identification of the major (> 10 %) metabolites in the excreta;
except if a minor (< 10 %) metabolite would appear to be of toxi-
cological concern,
— distribution of labelled material in tissues and products following a
single dose administration to animals already in steady state equili-
brium achieved with unlabelled additive.
The studies mentioned in 4.1.3.1 and 4.1.3.2 should include isotope
tracer or alternative relevant methods.
4.1.3.3. Study of the residues
— identification of those residues (parent compound, metabolites,
degradationproducts, boun d residues(1)) which represent more
than10 % of the total residue (except if a min or metabolite would
appear to be of toxicological concern) in the edible tissues and
products (milk, eggs) at metabolic equilibrium, i.e. following
multi-dose administration of the labelled substance; ratio of the
marker residue to total residues,
— kinetic study of the residues in the tissues (including milk and eggs
when appropriate) during the depletion period following achieve-
ment of steady-state and using the highest level proposed
metabolic profiling, identification of the target tissue(2) and of the
marker residue,
— depletionstudy of the marker residue from the target tissues
(including milk and eggs if appropriate) after withdrawal of the
additive following its repeated administration according to the
proposed conditions of use and sufficient to have reached steady-
state, inorder to set a withdrawal period onthe basis of the fixed
MRL,
— the withdrawal period for the additive must not be less than the time
necessary for the concentration of the marker residue determined in
the target tissue to fall below the MRL value (95 % confidence
limit). Spaced time points, suitably chosen by reference to the
depletion phase of the active substance and its metabolites, and at
least four animals per point depending on the species (size, genetic
variability) should be considered as a minimum requirement(3).1987L0153 — EN — 26.10.2001 — 003.001 — 12
(1) Bound residues correspond to the tissue residual fraction that is not extractable using
physico-chemical or biological means. They derive from the covalent binding of ametabolite of the compound with cellular macromolecules.
(
2) The target tissue is the edible tissue selected to monitor for the total residue in the
target animal.
(3) For the determination of a withdrawal period, the suggested minimum numbers of
healthy animals sampled at each slaughter or time point are the following:— lactating cattle, eight, including animals at second or subsequent lactations (four
high yielding cattle at an early stage of lactation and four low yielding cattle at a
late stage of lactation),
— other large animals, four per sampling time,— poultry, six per sampling time,— laying birds, ten eggs per time point,
— fish, tenper samplin g time.
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4.2. Studies on laboratory animals
These studies must be carried out with the active substance using inter-
nationally recognised standard test methods as described in the OECD
Guidelines for methodological details or in Directive 67/548/EEC, and
according to the principles of good laboratory practice (GLP). Addi-
tional studies on particular metabolites produced by the target species
may be necessary if these are not formed to a significant extent in the
laboratory test species. Also where there is data inmanthis may n eed
to be taken into consideration in deciding what additional studies toconduct.
4.2.1. Acute toxicity
Acute oral toxicity studies should be carried inat least two mammalian
species. One laboratory species may be replaced, if appropriate, by a
target species. It will not be necessary to identify a precise LD50; an
approximate determination of the minimum lethal dose is normally
adequate. In order to reduce the number and suffering of the animals
involved, the maximum dosage should not exceed 2 000 mg/kg body
weight and alternative methods (limit test, fixed dose method, acute
toxicity class method) are recommended.
Risks to workers should be assessed ina series of studies usin g the
product (active substan ce plus carrier inthe form inwhich it is to be
made available commercially). Studies onskinirritan cy must be
performed and if these give positive results, mucous membrane (e.g.
eye) irritancy should be assessed. Allergic potential — skin sensitisa-
tion potential should also be assessed. Acute inhalation studies should
be performed if the product is likely to form a respirable dust or mist.
4.2.2. Genotoxicity studies including mutagenicity
In order to identify active substances and, if appropriate, their metabo-
lites and degradation products with mutagenic and genotoxic properties,
a selected combination of at least three different genotoxicity tests must
be carried out. The test battery should normally include prokaryotic and
eukaryotic systems tests including mammalian in vitroandin vivotests
systems. If appropriate the tests should be performed without and with
mammalianmetabolic activation .
Reasons for the choice of the tests with regard to their reliability to
assess genotoxic effects on different genetic endpoints at the gene,
chromosome and genome level should be given. Additional tests may
be indicated depending on the outcome of the tests and taking into
consideration the whole toxicity profile of the substance as well as the
intended use. Tests must be carried out according to established and up-
to-date validated procedures. Whenthe test target is bon e marrow,
proof of exposure of the cells to the test substance is required in the
case of a negative result.
4.2.3. Subchronic (90-day) oral toxicity studies
The durationof the tests must be at least 90 days. For additives
intended for use in food producing animal species the studies should
be carried out on two animal species, of which one should be a non-
rodent species, which may be the target species. For additives intended
for use in animals not for human consumption the studies on the target
species are sufficient: the active substance must be administered orally
at least at three levels inadditionto a con trol group to obtaina dose
response.
The maximum dose should normally reveal evidence of harmful effects.
The lowest dose level should not produce any evidence of toxicity.
4.2.4. Chronic oral toxicity studies (including carcinogenicity studies)
A chronic toxicity study, which may include examination of carcino-
genicity, must be carried out in at least one rodent species.
Carcinogenicity studies may not be necessary if the active substance
and its metabolites:
— give consistently negative results in an appropriate range of geno-
toxicity tests,
— are not structurally related to known carcinogens, and— give no effects indicative of potential (pre)neoplasia in chronic toxi-
city assays.1987L0153 — EN — 26.10.2001 — 003.001 — 13
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4.2.5. Reproduction toxicity studies including teratogenicity
4.2.5.1. Two generation reproduction toxicity study
— Studies of reproductive function must be carried out and extend
over at least two filial generations (F1, F2) and may be combined
with a teratogenicity study. The substance under investigation shall
be administered to males and females at an appropriate time prior to
mating. Administration should continue until the weaning of the F2
generation.
— All relevant fertility, gestation, parturition, maternal behaviour,
suckling, growth and development of the F1 offspring from fertilisa-
tion to maturity and the development of the F2 offspring to weaning
must be carefully observed and reported.
4.2.5.2. Teratogenicity study
The teratogenicity study covers embryo and foetotoxicity. It must be
carried out inat least two species.
4.2.6. Metabolism and disposition studies
Studies on absorption, distribution in the body fluids and tissues, excre-
tion routes, must be performed. A metabolic study including the
metabolic balance and identification of the main metabolites in the
urine and faeces should be performed in animals of both sexes and the
same strains as those used in the toxicological studies. A single dose of
the labelled molecule (see 4.1.3) should be administered at steady state
equilibrium reached using the unlabelled compound at a dose similar to
the highest level proposed for use inthe target an imal.
4.2.7. Bioavailability of residues
The assessment of the risk to consumers related to certain residues
contained in animal products, namely bound residues, may take into
account an additional safety factor based on the determination of their
bioavailability using appropriate laboratory animals and recognised
methods.
4.2.8. Other specific toxicological and pharmacological studies
Further studies providing additional information useful for the assess-
ment of the safety of the active substance and its residues should be
conducted if there is any reason for concern.
4.2.9. Determination of a no observed effect level (NOEL)
All the above findings together with all the relevant published data
(including any suitable information on effects of the active substance
in human) and information, where appropriate, on closely related
chemical structures should be taken into consideration in identifying a
NOEL expressed as mg/kg body weight per day. The lowest NOEL
should be selected.
However the NOEL to be used for the calculationof the ADI should be
selected onthe basis of toxicological or pharmacological effects as
appropriate. For some additives, e.g. antibacterials, an ADI may be
better established onthe basis of effects onthe humangut microflora.
In the absence of internationally accepted and validated methods for
describing gut flora, effects on selected and sensitive human gut
bacterial strains may be more appropriate.
4.3. Safety evaluation for the human consumer
4.3.1. Proposal of the acceptable daily intake (ADI) for the additive
AnADI should where appropriate be proposed.
The ADI (expressed as mg of additive or additive related material per
personper day) is derived by dividin g the NOEL mg/kg body weight
by an appropriate safety factor and multiplying by a mean human
body weight of 60 kg. This NOEL expressed as mg/kg body weight
per day may be selected using toxicological or pharmacological find-
ings. In some cases an ADI based on the additives microbiological
properties may be more relevant. The choice will depend on which
property is most relevant in terms of health hazard to the consumer.
The safety factor used to determine the ADI for a particular additive
should be selected, bearing in mind the following:
— the nature of the biological effect used to identify the NOEL,
— the relevance of this effect to man and the reversibility of the effect,— the range and quality of the data used to identify the NOEL,1987L0153 — EN — 26.10.2001 — 003.001 — 14
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— any knowledge of the effect(s) of the residue constituents.
It is customary to employ a safety factor of at least 100 incalculatin g
the ADI (i.e. a factor of ten to allow for potential interspecies variation
and a further factor of ten to allow for possible differences in response
between individual humans). When data on the active substance are
available for humanbein gs a lower safety factor may be acceptable.
4.3.2. Proposal of the maximum residue limits (MRLs) of the additive
It is assumed in calculating the MRL that the intake of edible tissue,
milk and egg products is the sole source of potential human exposure.
If this is not the case an allowance must be made for other sources.
A number of these substances have been used as feed additives and for
other applications. In such cases the calculated MRLs would be
expected to be the same. There may also be instances where on strict
scientific considerations different MRLs are calculated for each use
when the route, amount, dosage frequency and duration of dosing, are
sufficiently different from those appropriate to use as feed additive, that
there are evidences indicating that the kinetics and/or metabolism may
result in different residue profiles. In such circumstances it is antici-
pated that the strictest MRL will be applied.
To establish anMRL the chemical n ature of the drug-related material
which is intended to be used to specify the tissue residue levels must
be defined. This is termed the marker residue. This residue constituent
must not necessarily be the toxicologically relevant residue but has to
be chosen as a suitable indicator to represent the total significant
residue. The ratios of the marker residue/total residues in connection
with the ADI (i.e. ratio of the marker residue/total radioactive residues,
marker residue/all biologically active residues) should be established at
all the time points during the depletion studies. In particular, this ratio
should be known at the time point retained to elaborate MRLs. A
suitable analytical method for this marker residue must also be avail-
able to ensure compliance with the MRL.
Inestablishin g MRLs (expressed as g/kg of marker residue per kg of edible wet tissue or
product) on the basis of an ADI, the following daily human food consumption values should
be applied:
Mammals Birds Fish
Muscle 300 g 300 g 300 g (*)
Liver 100 g 100 g
Kidney 50 g 10 g
Fat 50 g (**) 90 g (***)
+ Milk 1 500 g
+ Egg 100 g
(*) Muscle and skin in natural proportions.
(**) For pigs 50 g of fat and skin in natural proportions.
(***) Fat and skin in natural proportions.
The individual MRLs in different tissues should reflect the depletion
kinetics of the residues within those tissues in the animal species
intended for use. An analytical method with a limit of quantification
below the MRL is required (see sectionII poin t 2.5.3).
If a substan ce could result ina residue intissues an d produce, the
MRLs should be proposed insuch a way that the total amoun t of toxi-
cologically (or microbiologically) significant residue ingested(1) daily
should be lower thanthe ADI (see table above).
The MRL should be set only after consideration and inclusion of any
other potential sources of exposures of the consumer to components of
the residues.
For certainadditives residues could arise below the MRL values in
milk, eggs or meat which could nonetheless interfere with food quality
inparticular food processin g procedures, e.g. use of milk incheese
making. For such additives, it may be appropriate to consider a1987L0153 — EN — 26.10.2001 — 003.001 — 15
(1) Proposed calculation: (500 g meat (consisting of 300 g muscle, 100 g liver, 50 g
kidney, 50 g fat) or 500 g poultry (consisting of 300 g muscle, 100 g liver, 10 g kidney,
90 g fat) or 300 g fish) + 1500 g milk + 100 g egg.
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‘maximum (food product) processing compatible residue’ in addition to
establishing MRL values.
There are some circumstances where a MRL will not be required such
as:
— no bioavailability of the residues and no harmful effect on the
human gut including its microflora,
— complete degradation to nutrients or harmless substances in the
target species,
— ADI ‘not specified’ because of low toxicity in animal tests,
— where use is restricted entirely to feed for pet animals,— where a substance is also approved as a food additive(
1), a MRL is
normally not required if the marker residue is primarily the parentsubstance and it constitutes only an insignificant fraction of the
ADI of the food additive.
4.3.3. Proposal of the withdrawal period for the additive
The withdrawal period will be set onthe basis of the MRLs. The with-
drawal time comprises the period after cessationof the admin istration
of the proposed formulation of the additive which is necessary to enable
the residue levels to fall below the MRLs (95 % confidence limit).
To establish a withdrawal period a particular edible tissue may be iden-
tified as a surrogate for others, oftentermed the target tissue.
4.4. Worker safety assessment
Workers can be exposed mainly by inhalation or topical exposure while
manufacturing or handling or using the additive e.g. farm workers are
potentially exposed when handling or mixing the additive. Additional
information on how the substances are handled should be provided.
An assessment of risk to workers should be included.
Experience in the manufacturing plant is often an important source of
information in evaluating the risks to workers from exposure to the
additive itself by both airborne and topical routes. Of particular concern
are additives/additive-treated feed and/or animal excreta, which are in
or may give rise to, a dry powdery form and feed additives which
may have allergenic potential.
4.4.1. Toxicological risk assessment for worker safety4.4.1.1. Effects onthe respiratory system
Evidence should be provided that airborne levels of dust will not consti-
tute a hazard to the health of workers. This evidence should include
where necessary: inhalation tests in laboratory animals, published
epidemiological data and/or the applicants own data on its workplant
and/or irritancy and respiratory system sensitisation tests.
4.4.1.2. Effects onthe eyes an d skin
Where available, direct evidence of absence of irritancy and/or sensiti-
sation should be provided from known human situations. This should be
supplemented by findings from validated animal tests for skin and eye
irritation, and for sensitisation potential using the appropriate additive.
4.4.1.3. Systemic toxicity
The toxicity data generated to meet safety requirements (including
repeated dose toxicity, mutagenicity, carcinogenicity and reproductive
testing) should be used to assess other aspects of worker safety. In
doing so, it needs to be remembered that contamination of skin and/or
inhalation of the additive are the most likely routes of exposure.
4.4.2. Exposure assessment
Information should be provided on how the use of the additive is likely
to give rise to exposure by all routes — inhalation, through the skin or
by ingestion. This information should include a quantitative assessment,
where available, such as typical airborne concentration, dermal contam-
ination or ingestion. Where quantitative information is not available,
sufficient information should be given to enable an adequate assessment
of exposure to be made.1987L0153 — EN — 26.10.2001 — 003.001 — 16
(1) In accordance with Council Directive 89/107/EEC of 21 December 1988 on the approx-
imation of the laws of the Member States concerning food additives for use as
foodstuffs intended for human consumption (OJ L 40, 11.2.1989, p. 27).
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4.4.3. Measures to control exposure
Using the information from the toxicology and exposure assessment, a
conclusion should be drawn about the risks to health of the users
(systemic, toxicity, irritancy or sensitisation) when using measures to
control exposure, which are reasonable, in the circumstances. If the
risk is unacceptable, then precautionary measures should be taken to
control or to eliminate exposure. Product reformulation or modification
of the procedures for production, use and/or disposal of the additive are
preferred solutions. Use of personal protective devices should only be
regarded as a measure of last resort to protect against any residual risk
once control measures are in place.
4.5. Environmental risk assessment
Consideration of the environmental impact of feed additives is impor-
tant since the administration of feed additives is typically over a long
period (even for lifetime), large groups of animals may be involved
and many additives are poorly absorbed and therefore excreted intact
to a considerable extent. Nonetheless, in a number of cases, the need
for environmental assessment may be limited. It is inappropriate to set
strict rules in this general guideline. To assist in determining the envir-
onmental impact of a feed additive, a stepwise approach should be
followed (see decisiontree), where inthe first phase, additives which
do not need further testing can be clearly identified. For other additives
a second phase of studies (Phase IIA) are needed to provide additional
information, based upon which further studies (Phase IIB) may be
considered necessary. Studies, when applicable, should be conducted
according to Council Directive 67/548/EEC.
4.5.1. Phase I assessment
The purpose of Phase I assessment is to determine whether or not a
significant environmental effect of an additive or its metabolites is
likely, based largely ondata already established for other purposes.
Exemption from Phase II assessment may be made on one of two
criteria:
(a) The chemical nature and the biological effect of the additive and its
use indicate that impact will be negligible: i.e. where the additive
and/or its main (more than 20 % of the total residues in the excreta)
metabolite(s) are:
— physiological/natural substances (e.g. a vitamin, or a mineral)
that will not alter the concentration in the environment, unless
there is evident reason for concern (e.g. copper),
— additives intended for companion animals (excluding horses).
(b) The worst case predicted environmental concentration (PEC) is too
low to be of concern.
The worst case PEC for soil is likely to arise as a result of manure
generated during maximum excretion of the major residue constituents
(the additive and/or its major metabolites), being spread on land. The
PEC should be evaluated for each major residue constituent in manure
and for each compartment of concern. For the terrestrial compartment,
if the PEC does not exceed 100 µg/kg for the sum of the major residue
constituents in manure, or if the major residue constituents in manure
are readily degraded (degradationtime DT 50 < 30 days) (incase these
data are available) to natural constituents or to concentrations of less
than100 µg/kg, or if the PEC insoil (5 cm depth) is less thanµg/kg,
then no further assessment is required.
The worst case PEC for water may arise either from direct transfer of
spilled feed or excreta containing the additive and its metabolites into
water bodies or from the leaching of material in excreta or soil into
ground water. If the PEC for contamination of water bodies or ground
water is reliably estimated to be less than0,1 µg per litre n o Phase IIA
assessment of the environmental impact of the additive on the aqueous
compartment is necessary.
If the applicant cannot demonstrate that the proposed additive falls into
any of these exemption categories or when the additive is directly
released in the environment (e.g. aquaculture), Phase II assessment
will normally be required.1987L0153 — EN — 26.10.2001 — 003.001 — 17
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ENVIRONMENTAL RISK FROM FEED ADDITIVES
Decisiontree PhaseI
1987L0153 — EN — 26.10.2001 — 003.001 — 18
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4.5.2. Phase II assessment
Phase II assessment is in two parts: Phase IIA and Phase IIB.
The bioaccumulation potential of the additive and/or its main metabo-
lites and its influence on the predicted safety margin should be
assessed. Bioaccumulation is not considered to be potentially significant
if e.g. Kow(partitioncoefficien t) is < 3. Appropriate Phase IIB studies
will generally be needed if such safety margins cannot be established.
4.5.2.1. Phase IIA
The purpose of Phase IIA assessment is to identify risk to the environ-
ment by:
— refining the calculation of the PEC(s),
— determining the relationship between exposure, the levels of addi-
tive and/or major metabolites and short term adverse effects in
relevant surrogate animal and plant species for the environmental
compartment(s) of concern,
— using these findings to determine the predicted no effect concentra-
tion(s) (PNEC) value(s).
The following sequential procedure is recommended to determine the
risk:
(a) If not already completed in Phase I, a more refined PEC should be
calculated for each environmental compartment of concern. In
ascertaining the PEC account should be taken of:
— the concentration of additive and/or its major metabolites in
manure following administration of the additive to animals at
the proposed dose level. This calculation should include consid-
erationof excreta volumes an d dosage rates,
— the potential dilution of the excreted additive related material
due to normal manure processing practice and storage prior to
its applicationto lan d,
— the adsorption/desorption of the additive and its metabolites
onto soil, the persistence of residues in soil (DT50and DT90);
sediment in case of aquaculture,
— other factors such as photolysis, hydrolysis, evaporation, degra-
dationinsoil or water sedimen t systems, dilutionthrough
ploughing etc.
The highest value for the PEC obtained by these calculations for
each environmental compartment of concern should be adopted for
level IIA risk assessment purposes.
If a high persistence in soil (DT90> 1 year) at concentrations in
excess of 10 g/kg soil is anticipated at steady state, a level IIB
assessment may be needed.
(b) The levels producing serious short-term adverse effects for various
trophic levels in the environmental compartments of concern (soil,
water) must next be determined. These tests should follow
OECD(1) or similar well-established guidelines. Suitable tests for
the terrestrial environment include: toxicity to earthworms (50 %
lethal concentration, LC50value), phytotoxicity (50 % effective
concentration, EC50value) interrestrial plan ts, effects onsoil
micro-organisms (e.g. EC50for effects on methanogenesis and
nitrogen fixation). For the aquatic environment: fish: a 96-hour
LC50study; Daphnia magna : a 48-hour EC50study; algae: anLC50study and a toxicity study for sediment organisms.
(c) Calculation of the PNEC value for each compartment of concern
should be carried out. This is derived normally by taking the lowest
value observed (i.e.: the result inthe most sen sitive species) for an
adverse effect in the above ecotoxicity tests and dividing by a
safety factor of at least 100 depending on the indicator and number
of test species used.
(d) The calculated PEC and PNEC values should be compared. The
acceptable ratio of the PEC to the PNEC value will depend on the
nature of the test result used to determine the PNEC. Normally it
will be between 1 and 0,1. If significantly lower ratios than these
are identified, further ecotoxicological tests are unlikely to be1987L0153 — EN — 26.10.2001 — 003.001 — 19
(1) OECD Guidelines for Testing of Chemicals.
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necessary unless bioaccumulation is expected. Conversely, higher
ratios will require some Phase IIB testing.
4.5.2.2. Phase IIB (more detailed toxicological studies)
For those additives where, following Phase IIA assessment, doubt
remains regarding their environmental impact, more detailed studies
are required of the effects on biological species in the environmental
compartment(s) in which Phase IIA studies indicate possible concern.
In this situation, further tests are needed to determine the chronic and
more specific effects on appropriate animal, plant and microbial
species. It may be that inthe Phase IIA assessmen t the PEC value has
been over estimated. To demonstrate this it may be necessary to carry
out measurements of the environmental concentrations and, of the
persistence of the additive and/or its major metabolites in field use
situations.
Suitable additional ecotoxicity tests are described in a number of publi-
cations, e.g. in OECD guidelines. Three categories of environmental
species may need to be considered, animals, plants and micro-organ-
isms. Careful choice of such tests is necessary to ensure that they are
appropriate to the situationinwhich the additive an d/or its metabolites
may be released and dispersed in the environment.
The assessment of the impact on the terrestrial compartment may
include:
— a sublethal study of the effects onearthworms, further studies of the
impact onsoil microflora, phytotoxicity tests ona ran ge of econ om-
ically important plant species, studies on grassland invertebrates
including insects and feral birds,
—NB:a separate evaluation of mammalian toxicity may not be neces-
sary, as this aspect is likely to be addressed by mammaliantoxicity
testing conducted to determine the ADI.
The assessment of the impact on the aquatic compartment may:
— include chronic toxicity testing on the most sensitive aquatic organ-
isms identified in the Phase IIA assessment, e.g.: the fish early life
stage test, the Daphnia reproduction test, 72-hour algae tests and a
bioaccumulationstudy,
— include, where an adequate safety margin between the PEC and
PNEC values cannot be established, identification of effective miti-
gating measures to limit the environmental impact must be
provided.
5.SectionV:Formofmonograph
5.1. Identity of the additive
5.1.1. Proposed proprietary name(s).5.1.2. Type of additive according to its main function. Any other uses of the
active substance should be specified.
5.1.3. Qualitative and quantitative composition (active substance, other
components, impurities, batch to batch variation). If the active
substance is a mixture of active components, each of which is clearly
definable, the main components must be described separately and the
proportions in the mixture given.
5.1.4. Physical state, particle size distribution, particle shape, density, bulk
density; for liquids: viscosity, surface tension.
5.1.5. Manufacturing process including any specific processing procedures.5.2. Specifications concerning the active substance
5.2.1. Generic name, chemical name according to IUPAC nomenclature, other
generic international names and abbreviations. Chemical Abstracts
Service Number (CAS).
5.2.2. Structural formula, molecular formula and molecular weight. Qualita-
tive and quantitative composition of the main components, microbial
origin (name and place of culture collection where the strain is depos-
ited), if the active substance is a fermentation product.
5.2.3. Purity
Qualitative and quantitative composition of the active substances and
occurring accompanying impurities and toxic substances, confirmation
of the absence of the production organisms.1987L0153 — EN — 26.10.2001 — 003.001 — 20
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5.2.4. Relevant properties
Physical properties of the chemically specified substances: dissociation
constant, pKa, electrostatic properties, melting point, boiling point,
density, vapour pressure, solubility in water and organic solvents, Kowand Koc, mass and absorption spectra, NMR data, possible isomers and
any other appropriate physical properties.
5.3. Physico-chemical, technological and biological properties of the addi-
tive
5.3.1. Stability of the additive on exposure to environmental conditions such
as light, temperature, pH, moisture and oxygen. Proposal of a shelf life.
5.3.2. Stability during the preparation of premixtures and feedingstuffs, in
particular stability to anticipated process conditions (heat, moisture,
pressure/shear and time). Possible degradation or decomposition
products.
5.3.3. Stability during the storage of premixtures and processed feedingstuffs
under defined conditions. Proposal of a shelf life.
5.3.4. Other appropriate physico-chemical, technological or biological proper-
ties such as dispersability under favourable conditions in order to obtain
and keep homogeneous mixtures in premixtures and feedingstuffs, anti-
dusting and antistatic properties, dispersability in liquids.
5.4. Control methods
5.4.1. Descriptionof the methods used for the determin ationof the criteria
listed under items 2.1.3, 2.1.4, 2.2.3, 2.2.4, 2.3.1, 2.3.2, 2.3.3 and 2.3.4.
5.4.2. Description of the qualitative and quantitative analytical methods for
determining the marker residue of the active substance in target tissues
and animal produce.
5.4.3. If the said methods have beenpublished the literature referen ces may
suffice and the corresponding reprints should be given.
5.4.4. Information on the optimum storage conditions for the reference stan-
dards.
5.5. Biological properties of the additive
5.5.1. Particulars of the prophylactic effects for coccidiostats and other medic-
inal substances (e.g. morbidity, mortality, oocyst count and lesion
score).
5.5.2. For zootechnical additives other than those listed in 5.5.1 particulars of
the effects on feed intake, body weight, feed efficiency, product quality
and yield and any other parameter of benefit to the animal, the environ-
ment, the producer or the consumer.
5.5.3. For technological additives, relevant technological effects.
5.5.4. Any adverse effects, contra-indications or warnings (target animal,
consumer, environment), including biological interactions, with particu-
lars of their justification. Any ADI or MRLs established for other uses
of the active substance should be specified.
5.6. Details of the quantitative and qualitative residues in target tissues, if
any, found in animal produce following envisaged use of the additive
5.7. The ADI, the established MRLs and the withdrawal period should be
given, if appropriate
5.8. Other characteristics suitable for identification of the additive
5.9. Conditions of use
5.10. Date1987L0153 — EN — 26.10.2001 — 003.001 — 21
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6.SectionVI:Formofidentification note
1.Identity of the additive
1.1. Type of additive1.2. Physical state1.3. Qualitative and quantitative composition
1.4. Method of analysis of the additive and the residues
1.5. Community registration number (EC number)1.6. Packaging2.Specifications concerning the active substance
2.1. Generic name, chemical name, CAS Number
— Generic name
— Chemical name (IUPAC)— CAS number
2.2. Empirical formula
3.Physico-chemical, technological and biological properties of the
additive
3.1. Stability of additive
3.2. Stability during the preparation of premixtures and feedingstuffs
3.3. Stability during storage of premixtures and feedingstuffs3.4. Other properties4.Conditions of use
4.1. Species or category of animals, maximum age if specified
4.2. Minimum and maximum content in feedingstuffs4.3. Contra-indications, interactions4.4. Warnings
5.Person responsible for putting into circulation
5.1. Name5.2. Address5.3. Registrationn umber
6.Manufacturer
6.1. Name6.2. Address6.3. Approval number or registration number assigned to the establish-
ment or the intermediary.
7.Date
7.SectionVII:Renewalofauthorisationofadditiveswhoseauthorisa-
tion is linked to a person responsible for putting them into
circulation
1.General
An updated dossier and monograph should be prepared according to
the most up-to-date guidelines and a list provided of all variations of
any type since the authorisation for putting into circulation or the
last renewal.
It must be confirmed that the monograph and safety file has have
been adapted to include all new information relevant to the additive
or now required as a result of changes in these guidelines.
Information must also be provided on the authorisation status world
wide and sales volume.1987L0153 — EN — 26.10.2001 — 003.001 — 22
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2.Identity of the active substance and of the additive
Evidence should be presented to show that the additive has not been
changed or altered in composition, purity or activity in respect of the
additive authorised. Any change of the manufacturing process should
be reported.
3.Efficacy
Evidence should be presented to show that the additive retains the
claimed efficacy under conditions of animal production current in
the European Union at the time of application for renewal of the
authorisation. This should include an account of general experience
with the use of the additive and performance monitoring.
4.Microbiology
Special regard should be given to possible development of resistance
to antimicrobials during the long-term use under practical conditions.
The tests must thus be performed under field conditions in farms,
where the additive has been routinely used for as long a time as
possible. A selection of common intestinal bacteria should be used
as test organisms, and the selection should include relevant endo-
genous and exogenous gram-positive as well as gram-negative
organisms.
If the tests show a change in the resistance pattern compared to the
original figures, the resistant bacteria must be examined for cross-
resistance to relevant antibiotics used for treatment of infectious
diseases in man and animals. The most important are antibiotics
belonging to the same group as the additive, but also other groups
of antibiotics should be included in the trial.
Results of appropriate monitoring programmes should be reported.
5.Safety
Evidence should be presented that in the light of the current knowl-
edge the additive remains safe under the approved conditions for
target species, consumers, operators and the environment. A safety
update for the period since the authorisation for putting into circula-
tion or the last renewal with information on the following items
should be presented:
— reports on adverse effects including accidents (previously
unknown effects, severe effects of any type, increased incidence
of known effects) for target animals, operators and the environ-
ment. The report on adverse effect should include the nature of
the effect, number of affected individuals/organisms, outcome,
conditions of use, causality assessment;
— reports on previously unknown interactions and cross-contamina-
tions;
— data from residue monitoring where appropriate;— any other information concerning the safety of the additive.
If no further information is provided on any of these factors, the
reasons for this should be clearly identified.
8.SectionVIII:Newapplicantrelying onthefirstauthorisationofan
additive whose authorisation is linked to a person responsible for
puttingthemintocirculation
Since reliance can be placed on the evaluation of the data supplied for
initial authorisation a dossier prepared in relation to an application
under Article 9c(3) need comply only with the following requirements.
An additive can be considered as identical for this purpose if the quali-
tative and quantitative composition and the purity of active and inactive
components are essentially similar, the preparation is the same and the
conditions of use are identical.
For such products it will normally not be necessary to repeat pharmaco-
logical, toxicological and efficacy studies and an abridged application
canbe submitted. This must in clude expert reports.
— A complete Section II and a monograph must be submitted.
— Data must be provided indicating that the specification range of the
physical, chemical characteristics of the additive is essentially
similar to that of the established product.
— It must be confirmed that further scientific knowledge in the avail-
able literature on the additive has not changed the original1987L0153 — EN — 26.10.2001 — 003.001 — 23
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assessment on efficacy since the authorisation for putting into circu-
lationof the origin al additive.
— Special regard should be givento possible developmen t of resis-
tance to antimicrobials during the long-term use of the active
substance under practical conditions. The tests must thus be
performed under field conditions in farms, where the active
substance has been routinely used for as long a time as possible. A
selection of common intestinal bacteria should be used as test
organisms, and the selection should include relevant endogenous
and exogenous gram-positive as well as gram-negative organisms.
— If the tests show a change in the resistance pattern compared to the
original figures, the resistant bacteria must be examined for cross-
resistance to relevant antibiotics used for treatment of infectious
diseases in man and animals. The most important are antibiotics
belonging to the same group as the additive, but also other groups
of antibiotics should be included in the trial.
— Evidence should be presented that in the light of the current scien-
tific knowledge in the available literature the additive remains safe
under the approved conditions for target species, consumers, opera-
tors and the environment.
— The conformity of the withdrawal period with the MRL has to be
established.
PART II
MICRO-ORGANISMSANDENZYMES (1)1987L0153 — EN — 26.10.2001 — 003.001 — 24
(1) See CommissionDirective 94/40/EC (OJ L 208, 11.8.1994, p. 15), as amen ded by
Directive 95/11/EC.