ARCHIVESHISTORIQUES
DELACOMMISSION
COLLECTION RELIEEDES
DOCUMENTS "COM"
COM(82)615
Vol.1982/0193

Disclaimer
Conformément aurèglement(CEE,Euratom)n°354/83duConseildu1erfévrier1983
concernantl'ouvertureaupublicdesarchiveshistoriquesdelaCommunauté économique
européenne etdelaCommunauté européenne del'énergieatomique(JOL43du15.2.1983,
p.1),telquemodifiéparlerèglement(CE,Euratom)n°1700/2003du22septembre2003
(JOL243du27.9.2003,p.1),cedossierestouvertaupublic.Lecaséchéant,lesdocuments
classifiésprésentsdanscedossierontétédéclassifiés conformément àl'article5dudit
règlement.
Inaccordance withCouncilRegulation(EEC,Euratom)No354/83of1February1983
concerningtheopeningtothepublicofthehistoricalarchivesoftheEuropeanEconomic
Community andtheEuropeanAtomicEnergyCommunity (OJL43,15.2.1983,p.1),as
amendedbyRegulation(EC,Euratom)No1700/2003of22September2003(OJL243,
27.9.2003,p.1),thisfileisopentothepublic.Wherenecessary,classifieddocuments inthis
filehavebeendeclassified inconformitywithArticle5oftheaforementioned regulation.
InÜbereinstimmung mitderVerordnung(EWG,Euratom)Nr.354/83desRatesvom1.
Februar1983überdieFreigabederhistorischen ArchivederEuropäischen
Wirtschaftsgemeinschaft undderEuropäischen Atomgemeinschaft (ABI.L43vom15.2.1983,
S.1),geändertdurchdieVerordnung (EG,Euratom)Nr.1700/2003vom22.September2003
(ABI.L243vom27.9.2003,S.1),istdieseDateiderÖffentlichkeit zugänglich.Soweit
erforderlich,wurdendieVerschlusssachen indieserDateiinÜbereinstimmung mitArtikel5
dergenanntenVerordnungfreigegeben.

COMMItMQN OFTHEEUROPEAN COMMUNITIES
COM(82)615final
Brussels,1October1982
Proposalfora
COUNCILDIRECTIVE
onthefixingofguidelinesfortheassessmentofcertainproductsused
inanimalnutrition
(submittedtotheCouncilbytheCommission )
C0M(S2>615final

l-h
EXPLANATORY MEMORANDUM
On30June1982,theCounciladoptedDirective82/471/EECconcerningcertain
productsusedinanimalnutritiononaccountoftheirproteincontent.
Thepurposeofthatframeworkdirectivewastoauthorize-undercertain
conditions-theproduction anduseofbioproteinsforanimalfeeding.
Moreparticularly,thepurposewastocontrolnotonlytheuseofcertain
well-knownproductssuchasaminoacids,non-protein nitrogenous compounds
andcertainyeasts,butabovealltheuseofnovelproteinsobtainedfrom
algae,fungiandthecultureofmicro-organisms (bacteria,yeast)on
substratesofvariousoriginssuchaspetroleumormethanol.
Inordertoestablisheverypossibleguaranteeensuringthesafeguardof
humanandanimalhealthandprotectionoftheenvironment,theCouncil
decidedtomakeproteinproductsobtainedfrombacteria,yeasts,algae
andinferiorfungisubjecttoaverysevereauthorization procedure,
underwhichtheCommissionwillberequiredtoconsulttheScientific
CommitteeforAnimalNutritionandtheScientific CommitteeforFood,and
toobtaintheopinionoftheStandingCommitteeforFeedingstuffs.
Furthermore,toensurecompliancewiththeprovisions onhumanand
animalhealthintheframeworkDirective,Article7providesthat,for
eachproduct,theMemberStateconcernedmustsendadossierofficially
toalltheotherMember"StatesandtotheCommissionandtothemembers
oftheScientific Committees .Thisdossiermustbedrawnupinaccor­
dancewithguidelines whichtheCouncilundertooktoadoptintimefor
themtoapplyasfromthedateofapplication ofthebasicDirective.
i

-2-
Accordingly,thepurpospofthepresentDirectiveistoLaydownthesaid
guidelinesforthepresentation oftheproductdossiersreferredtoin
Article7ofDirective82/471/EEC.Theseguidelineswillbeusedasa
referenceforassessingtheproductsconcernedinthelightofcurrent
scientific knowledgeandforensuringthatsuchproductsmeetthebasic
criterialaiddownfortheirauthorization .

Proposalfora
COUNCILDIRECTIVE
onthefixingofguidelinesfortheassessmentofcertainproductsused
inanimalnutrition
THECOUNCILOFTHEEUROPEANCOMMUNITIES,
HavingregardtotheTreatyestablishing theEuropeanEconomicCommunity,
HavingregardtoCouncilDirective82/471/EECof30June1982concerningcertain
1productsusedinanimalnutrition,andinparticularArticle7thereof,
HavingregardtotheproposalfromtheCommission,
WhereasDirective82/471/EECprovidesthattheproductsbelongingtocertain
groupsmustbeexaminedonthebasisofadossierforwardedofficiallyto
theMemberStatesandtheCommission;
Whereassuchdossiersmustmakeitpossibletoverifythattheproductsinquestion
complywiththegeneralprincipleslaiddownintheDirectiveinrespectofthe
inclusionofnewproductsintheAnnex;
Whereasithasbeenfoundnecessarytoprovideforthedossierstobecompiledin
accordancewithcommonguidelinesdefining,foreachprinciple,thescientific
datawhichmakeitpossibletoidentifyandcharacterize theproductsconcerned
andthestudiesnecessaryinordertoevaluatetheirnutritionalproperties
andbiologicaleffects;whereastheseguidelinesmustbeapplicable onthedate
onwhichDirective82/471/EECitselfentersintoforce;
Whereastheguidelinesareintendedprimarilyasageneralguide;whereas,
dependingonthenatureoftheproductoritsconditionsofuse,theextentof
thestudiesnecessaryinordertoevaluateitspropertiesoritseffectsmay
vary;
Whereastheguidelineshavebeendrawnuponthebasisofpresentscientificand
technicalknowledgeandtheymaybeadaptedifnecessarytoanydevelopments in
thissphere,
– ./.
(1)J.O.N°L213,21.7.1982,p.8.

HASADOPTEDTHISDIRECTIVE :
Article 1
MemberStatesshallprescribethatthedossiersontheproductslistedinpoints
1.1.and1.2.oftheAnnextoDirective82/471/EECaretobecompiledin
accordancewiththeguidelinessetoutintheAnnexhereto.
Article2
MemberStatesshallbringintoforceon13July1984thelaws,regulationsor
administrative provisionsnecessaryinordertocomplywiththisDirective.They
shallforthwithinformtheCommissionthereof.
Article3
ThisDirectiveisaddressedtotheMemberStates.
DoneatBrussels, FortheCouncil

ANNEX
GUIDELINES FORTHEASSESSMENT OFCERTAINPRODUCTS
USEDINANIMALNUTRITION
GENERALASPECTS
Thisdocumentisintendedasaguideforestablishing dossiersonproducts
Listedinitem1oftheAnnexofCouncilDirective82/471/EECof30June
1982concerningcertainproductsusedinanimalnutrition (1),whichhave
beenobtainedfromculturingmicroorganisms andwhichareproposedasa
newsourceofproteinsinanimalnutrition.Thesedossiersshouldenable
anassessmentofsuchproductsbasedonthepresentstateofknowledgeand
shouldensuretheircompliancewiththefundamentalprincipleslaiddown
forpermittingtheiruse,whicharethesubjectoftheprovisionsofArticle
6(2)oftheabovementionedDirective.
Allthestudiesoutlinedinthisdocumentmayberequiredand,ifnecessary,
additionalinformation mayberequested.Asageneralrule,alltheinforma­
tionnecessarytoestablishtheidentityofthemicroorganism andthecompo­
sitionoftheculturemedium,andalsothemanufacturing process,characte­
ristics,presentation,conditionsofuse,methodsofdetermination andnutri­
tionalpropertiesoftheproductmustbeprovided.Thesameappliestothe
informationnecessarytoassessthetoleranceoftheproductbythetarget
speciesandtherisksformanandtheenvironment,whichcouldresultdirectly
orindirectlyfromtheuseoftheproduct.Thetoxicological studiesrequired
forthispurposewilldependonthenatureoftheproduct,theanimalspecies
concernedandthemetabolismoftheproductinlaboratoryanimals.
Thedocumentation tobeprovidedshouldincludedetailedreports,presented
intheorderandwiththenumberingproposedintheseguidelinesandshould
beaccompaniedbyasummary.Theomissionofanyproposedstudiesshouldbe
justified.Thepublicationsquotedasreferencesshouldbeattached.
OBSERVATIONS
Theterm"product",asusedintheseguidelines,referstoanyproteinaceous
productinthestateinwhichitwillbepresentedasfeedingstufforcomponent
ofafeedingstuff .
Anymodificationinthemanufacturingprocessorintheconditionsofuseof
aproductwillrequirenotification and,ifnecessary,additionaldocumentation
foranewassessment .
Theguidelineswillbeupdatedasnewscientificknowledgedevelops.
(1)0JNoL213,21.7.1982,p.8

PRESENTATION OFSTUDIES
I. Microorganism ,cultur-s medium andmanufacturing process ,charac­
teristics ofproduct ,presentation andconditions ofuse,methods
ofdetermination .
II.Studies onthenutritional properties oftheproduct .
III .Studies onthebiological consequences oftheuseoftheproduct in
animal nutrition .
IV.Other relevant studies .
SECTION I: MICROORGANISM ,CULTURE MEDIUM ANDMANUFACTURING PROCESS ,
CHARACTERISTICS OFPRODUCT ,PRESENTATION AND CONDITIONS
OFUSE,METHODS OFDETERMINATION
1. MICROORGANISM
1.1 .Classification ,morphology ,biological properties ,anygenetic
manipulation .
1.2 .Innocuity ,possible survival outside thefermenter andenviron -
mental conséquences .
1.3 .Constancy andpurity ofstrains cultivated .Methods used tocheck
these criteria .
2. CULTURE MEDIUM AND MANUFACTURING PROCESS
2.1 .Composition ofsubstrate ,added substances ,etc .
2.2 .Manufacturing andpurification processes .Methods used tocheck
theconstancy ofcomposition oftheculture product andthe
detection ofanychemical andbiological contamination during
production .
2.3 .Technical processes ofpreparation foruse .
3. CHARACTERISTICS OFPRODUCT
3.1.Physical andphysico-chemical properties :macro -andmicro -
morphology ,particle size ,density ,specific weight ,hygrosco­
picity ,solubility ,solution characteristics (pH,rheological
characteristics ),electrostatic properties ,etc .
3.2 .Chemical composition :
3.2.1 .Content ofmoisture ,crude protein ,crude fat,crude fibre ,
ash,nitrogen-free extract .Limits ofvariation .
3.2.2 .Content oftotal ,ammonium ,amide ,nitrate andnitrite nitrogen ,
true protein .Qualitative andquantitative composition oftotal
andfree amino acids ,nucleic acids (purine andpyrimidine bases ).

3.2.3.Qualitative andquantitative compositionoftotallipids:fatt'y
acids,non-saponifiablématter,lipidsolublepigments,etc.
3.2.4.Qualitative andquantitative compositionofcarbohydrates andrelatedsubstances .
3.2.5.Qualitative andquantitative compositionofinorganiccomponents.
3.2.6.Qualitative andquantitative colpositionofvitamins.
3.2.7.Qualitative andquantitative compositonoftheotherconsti­
tuents:additives,residuesofsubstrateandsolvents,conta­
minants(inparticular,polycyclicaromatichydrocarbons,nitro­
samines),etc.
3.3.Contamination bymicroorganisms duringthemanufacturing process.
3.4.Behaviourandstabilityoftheproduct,assuchandwhenmixed
withfeedingstuffsincurrentuse,duringstorage.
4.PRESENTATION ANDCONDITIONS OFUSE
4.1.Proposednamesformarketingtheproduct.
4.2.Proposedformulations formarketingtheproduct.
4.3.Intendeduseoftheproductinanimalnutrition.Proposedcon­
centrations incompleteandcomplementary feedingstuffsforthe
animalspeciesconcerned.
5.METHODS OFDETERMINATION
Qualitative andquantitative methodsfordetermination ofthe
productincompleteandcomplementary feedingstuffs.
N.B.:Description ofthesemethodsshouldbeaccompanied byin­
formation astospecificity,sensitivity,limitsofde­
tection,marginoferror,possibleinterferences byother
substances .Samplesoftheproductinitsvariousproposed
presentations shouldbeavailable.
SECTIONII:STUDIES ONTHENUTRITIONAL PROPERTIES OFTHEPRODUCT
1.ASSESSMENT OFPROTEINVALUE
1.1.Chemicaland/ormicrobiological studies.
1.2.Studiesonlaboratoryanimals(preferablyrats):digestibility ,
proteinefficiencyratio(PER),biologicalvalueoftheproduct
comparedwithreferenceproteins.
2.STUDIESONTARGETSPECIES
Thefollowingstudiesshouldbeperformedoneachtargetspecies
incomparisonwithacontrolgroupreceiving,underthesamecon­
ditionsofnutritional balance,adietincurrentusecontaining
equivalent amountsofproteinnitrogen.

t-.1.Proteinndenergy .up,»let.ieiitdticn valueoft;ieproductinthe
reticvisi.nc?rthe>.conditionsofuseatvariousphysio­
logicalstagesofth.animals(growingperiod,pregnancy,laying,
etc.).
2.2.Influenceoftheproductundertheproposedconditionsofuseon
growthret*,feedconversionrate,morbidity,mortality,etc.
2.3.Optimumlevelsofincorporation oftheproductintherations.
2.4.Effectoftheproductundertheproposedconditionsofuseonthe
composition andonnutritive ,,technological andorganoleptic
qualityofneat,offal,eggsandmilk.
3.EXPERIMENTAL CONDITIONS INTHESTUDIESONTARGETSPECIES
Giveadetaileddescription ofthetestsperformedandprovide
thefollowingdata:
3.1.Species,breed,agea.idsexoftheanimals,identification
procedure .
3.2.Numberoftestandcontrolgroups;numberofanimalsineach
group(thenumbershouldbelargeenoughforstatistical analysis
usingsuitablestatistical parameters).
3.3.Levelsofincorporation oftheproduct,qualtitative andquan­
titativecomposition oftherationanditsanalysis.
3.4.Locationofeachexperiment,physiological stateandanimal
healthconditions,rearingconditions (theseshouldreflectthose
usedinpracticeintheCommunity).
3.5.Exactdurationoftestinganddateoftheanalysesperformed.
3.6.Adverseeffectswhichoccuredduringtheexperiment andtimeof
theirappearance .
SECTIONIII:STUDIES CONCERNING THEBIOLOGICAL CONSEQUENCES OFTHE
USEOFTHEPRODUCTINANIMALNUTRITION
Thestudiesoutlinedinthissectionareintendedtopermitassessment
ofthesafetyinuseoftheproductinthetargetspecies,andofthe
risksformanandtheenvironment whichcouldresultdirectlyorin­
directlyfromthisuse.Thetoxicologicalstudiesrequiredforthis
purposewilldependonthenatureoftheproduct,theanimalspecies
concerned andthemetabolism oftheproductinlaboratory animals.
1.STUDIES ONTARGETSPECIES
Thefollowingstudiesshouldbeperformed oneachtargetspecies
incomparison withacontrolgroupreceiving,underthesamecon­
ditionsofnutritional balance,adietincurrentusecontaining
equivalent amountsofproteinnitrogen.

–5-
1.1.Maximumincorporation ratesoftheproductintherationwithout
producinganyadverseeffect.
1.2.Effectsofingestionoftheproductundertheproposedconditions
ofuseonmicroorganisms ofthefloraofthealimentarytractand
oncolonization ofpathogensinthealimentarytract.
1.3.Investigation undertheproposedconditionsofuseofpossible
residuesoftheproduct(substrate,culturemedium,solvents,
contaminants )inanimalproducts(meat,milk,eggs,etc.).
1.4.Investigation undertheproposedconditionsofuseofpossible
residuesoftheproduct(substrate,culturemedium,solvents,
contaminants )inexcreta.
2-STUDIESONLABORATORY ANIMALS
2.1.Metabolism
Fateoftheproductintheanimal:absorption,elimination,etc.
2.2.Mutagenicity
Investigations ofpotentialmutagenicity duetocontaminants (in
particularmycotoxins)orresiduesoftheproduct(substrate,
culture,medium,solvents),includinginvitroscreeningtests
usingmetabolicactivation systems.
2.3.Toxicologicalstudies
Thefollowingstudiesshouldbeperformedincomparisonwith
controlgroupsreceiving,underthesameconditionsofnutriti­
onalbalance,adiet-incurrentusecontainingequivalent amounts
ofproteinnitrogen.Toxiceffectsshouldbeinvestigated toelu­
cidatetheircauseandmechanisms andtoascertainthattheydo
notresultfromnutritional imbalanceorfromanoverdosageof
theproductinthediet.
2.3.1.Subchronictoxicity(atleast90days)
Ingeneral,thesestudiesshouldbecarriedoutontwoanimal
species,oneofwhichbeingarodent.Theproductshouldbe
administered inthedailyrationinatleasttwolevelsofin­
corporation .Ifpossible,theseshouldbechosensoastode­
termineano-effectlevelandalevelshowingsomeadverseeffect.
Theexperimental groupsshouldcontainanadequatenumberof
animalsofeachsex.Acontrolgroupshouldalwaysbeincluded.
Allrelevantbiological datashouldberecordedatappropriate
intervals,particularly dataongrowthrate,feedconsumption,
haematology,urineanalysis,biochemical parameters,mortality,
organweights,grosspathologyandhistopathology ofmajororgans
andtissues.Theresultsshouldbepresentedindetailand,as
faraspossible,shouldincludestatistical assessment .

<.3.2 „Chronic '(oxvcity
Ingeneral ,c.iror.v ,fxicity studies should becarried outontwo
animal species ,one ofwhich being arodent .Theproduct should
beadministered inrr)daily ration inatleart two levels of
incorporation .Expt ^ :;icnts should extend for ?.mininum oftwo
years Ir.th?rat oroJweeks inmice .Theexperimental groups
should contain anc....;qujte number ofanimals of?cch sex .A
control group shoulci always beincluded .Theexperiment ,if
continued beyond the !iinimum period ,should beterminated when
survival inanybutthegroup with thehighest level ofincorpo­
ration has fallen to20%.
The biological examinations mentioned under item 2.3.1 .should be
carried outpreferr^ly onasmall satellito group ofanimals at
appropriate inte'v^is throughout theexperiment andonthe sui–
viviny animals att.seend oftheexperiment .Forassessing
carcinogenicity ,particular attention should bepaid tothetime
ofappearance ,theVstological types ofanyobserved tumours and
their incidence .Any effect onthe incidence oftumours and/or
theincidence orprogress ofdiseases should beassessed by
reference tocontrol iroups ,asindicated inparagraph 2.3 .The
result should bepresented indetail and,asfaraspossible ,
should include statistical assessment .
2.4 .Other studies
Reproduction studies should extend over atleast two filial
generations andmay becombined with embryotoxicity including
teratogenicity studies .Particular attention should bepaid to
fertility ,fecundity andobservation onpost-natal development of
litters .
2.5 .Experimental conditions inthe studies onlaboratory animals
Give detailed description ofthe tests performed andprovide the
following data :
2.5.1 .Species ,breed ,strain andsexofanimals .
2.5.2 .Number oftest and control groups ,number ofanimals ineach
group (thenumber should belarge enough forstatistical analysis
using suitable statistical parameters ).
2.5.3 .Levels ofincorporation oftheproduct ,qualitative andquanti­
tative composition ofthe ration and itsanalysis .
2.5.4 .General rearing conditions throughout theperiod oftesting .
2.5.5 .Exact duration oftesting anddate ofexaminations performed .
2.5.6 .Rate andtiming ofdeaths for thevarious test groups .
2.5.7 .Pathological incidents which occurred during theexperiment and
time oftheir appearance .

-7–
3.STUDIES CONCERNING THEENVIRONMENT
Depending onthenatureofpossibleresiduesoftheproduct
(substrate,culturemedium,solvents,contaminants )inexcretaof
targetspecies,dataonthefateoftheseresiduesinmanure,
soilandwaterandalsotheireffectsonsoilbiology,plant
growthandaquaticlifemayberequired.
SECTIONIV:OTHERRELEVANT STUDIES
Depending onthenatureandtheconditions ofuseoftheproduct,data
onallergiceffects,onirritation oftheskinandmucousmembranesof
theeye,respiratory ordigestivetractmayberequiredtoassesspos­
siblerisksinhandlingtheproductandtopreventthem.