1997L0012 — EN — 31.12.1998 — 001.001 — 1
This document is meant purely as a documentation tool and the institutions do not assume any liability for its contents
►B COUNCILDIRECTIVE97/12/EC
of17March1997
amending and updating Directive 64/432/EEC on health problems affecting intra-Community
tradeinbovineanimalsandswine
(OJ L 109, 25.4.1997, p. 1)
Amended by:
Official Journal
No page date
►M1Council Directive 98/99/EC of 14 December 1998 L 358 107 31.12.1998
▼B
COUNCILDIRECTIVE97/12/EC
of17March1997
amending and updating Directive 64/432/EEC on health problems
affecting intra-Communitytradeinbovineanimalsandswine
THE COUNCIL OF THE EUROPEAN UNION,
Having regard to the Treaty establishing the European Community, andin particular Article 43 thereof,
Having regard to the proposal from the Commission(
1),
Having regard to the opinion of the European Parliament(2),
Having regard to the opinion of the Economic and Social
Committee(3),
Whereas important progress has been made in the harmonization of the
veterinary field, in particular through the adoption by the Council ofDirective 90/425/EEC of 26 June 1990 concerning veterinary andzootechnical checks applicable in intra-Community trade in certainlive animals and products with a view to the completion of the internal
market(
4), Directive 91/496/EEC of 15 July 1991 laying down the prin-
ciples governing the organization of veterinary checks on animalsentering the Community from third countries and amending Directives89/662/EEC, 90/425/EEC and 90/675/EEC(
5), Directive 85/511/EEC of
18 November 1985 introducing Community measures for the control offoot-and-mouthdisease(
6), and Directive 92/119/EEC of 17 December
1992 introducing general Community measures for the control of
certain animal diseases and specific measures relating to swine vesi-cular disease(
7);
Whereas the Council in its resolution 94/C 16/01 of 22 December 1993
on the strengthening of veterinary epidemiological surveillancemeasures(
8) agreed to do everything in its power to ensure rapid imple-
mentation of the principles of that resolution in the context of the
amendment of Directive 64/432/EEC of 26 June 1964 on health
problems affecting intra-Community trade in bovine animals andswine(
9);
Whereas it is necessary, in the light of this situation, to amend Direc-
tive 64/432/EEC in particular as regards the period of residence in aMember State prior to movement, rules for trade in animals under 15days old, rules for control of certain diseases and rules applicable toassembly centres, transporters and dealers;
Whereas for the purpose of rapid and accurate tracing of animals for
animal health reasons each Member State should create a computerized
database which shall record the identity of the animals, all holdings on
their territory and the movements of the animals;
Whereas Directive 64/432/EEC has been substantially amended on a
number of occasions; whereas for the sake of clarity that Directiveshould be updated,1997L0012 — EN — 31.12.1998 — 001.001 — 2
(1) OJ No C 33, 2. 2. 1994, p. 1.
(2) OJ No C 128, 9. 5. 1994, p. 105.
(3) OJ No C 133, 16. 5. 1994, p. 31.
(4) OJ No L 224, 18. 8. 1990, p. 29. Directive as last amended by Directive 92/
118/EEC (OJ No L 62, 15. 3. 1993, p. 49).
(5) OJ No L 268, 24. 9. 1991, p. 56. Directive as last amended by Directive 96/
43/EC (OJ No L 162, 1. 7. 1996, p. 1).
(6) OJ No L 315, 26. 11. 1995, p. 11. Directive as last amended by the 1994
Act of Accession.
(7) OJ No L 62, 15. 3. 1993, p. 69. Directive as last amended by the 1994 Act
of Accession.
(8) OJ No C 16, 19. 1. 1994, p. 1.
(9) OJ No 121, 29. 7. 1964, p. 1977/64. Directive as last amended by Directive
95/25/EC (OJ No L 243, 11. 10. 1995, p. 16).
▼B
HAS ADOPTED THIS DIRECTIVE:
Article 1
The Articles of and the Annexes B, C, D (Chapter II) and E to Direc-
tive 64/432/EEC shall be replaced by the text annexed to this Directive,
the Annexes A, D (Chapter I) and F shall be replaced by the text
annexed to Directive 98/46/EC.
Article 2
1. Member States shall bring into force the laws, regulations and
administrative provisions necessary to comply withth is Directive►
M1not later than 1 July 1999 ◄. They shall forthwith inform the
Commission thereof.
When Member States adopt these measures, they shall contain a refer-
ence to this Directive or shall be accompanied by such reference on the
occasion of their official publication. The methods of making such areference shall be laid down by the Member States.
2. Member States shall communicate to the Commission the texts of
the main provisions of domestic law which they adopt in the fieldgoverned by this Directive.
Article 3
This Directive shall enter into force on the day of its publication in the
Official Journal of the European Communities .
Article 4
This Directive is addressed to the Member States.1997L0012 — EN — 31.12.1998 — 001.001 — 3
▼M1
▼B
▼B
ANNEX
DIRECTIVE64/432/EEC
(amendmentandupdatingoftheArticlesandtheAnnexes)
Article 1
This Directive shall apply to intra-Community trade in bovine animals and
swine withth e exception of feral pigs as defined in Article 2 (e) of Directive
80/217/EEC(1) without prejudice to the provisions laid down in Directives 80/
215/EEC(2), 85/511/EEC, 88/407/EEC(3), 89/608/EEC(4), 90/425/EEC, 90/429/
EEC(5), 90/667/EEC(6), 91/496/EEC, 91/628/EEC(7), 92/102/EEC(8), 92/119/
EEC and Decision 90/424/EEC(9).
Article 2
1. The definitions given in Article 2 of Directive 90/425/EEC and Article 2
of Directive 91/628/EEC shall apply.
2. In addition the following definitions apply for the purposes of this Direc-
tive:
(a)herdmeans an animal or group of animals kept on a holding (within the
meaning of Article 2 (b) of Directive 92/102/EEC) as an epidemiological
unit; if more than one herd is kept on a holding, each of these herds shall
form a distinct unit and shall have the same health status;
(b)animal for slaughter means a bovine animal (including the species Bison
bisonandBubalus bubalus ) or swine intended to be taken to a slaughter-
house or assembly centre from which it may only move to slaughter;
(c)animals for breeding or production means bovine animals (including the
species Bison bison andBubalus bubalus ) and swine other than those
referred to in (b), including those intended for breeding, milk or meat
production, or draft purposes, shows or exhibition with the exception of
animals taking part in cultural and sporting events;
(d)officiallytuberculosis-free bovine herd means a bovine herd which satisfies
the conditions laid down in ►M1Annex A.I, paragraphs 1 and 2 ◄;
(e)officiallytuberculosis-free Member State orregion of a Member State
means a Member State or part of a Member State which satisfies the condi-
tions laid down in ►M1Annex A.I, paragraphs 4 and 5 ◄;
(f)officiallybrucellosis-free bovine herd means a bovine herd which satisfies
the conditions laid down in ►M1Annex A.II, paragraphs 1 and 2 ◄;
(g)officiallybrucellosis-free region means a region of a Member State which
satisfies the conditions laid down in Annex A.II, paragraphs 7, 8 and 9;
(h)officiallybrucellosis-free Member State means a Member State which satis-
fies the conditions laid down in ►M1Annex A.II, paragraphs 7, 8 and
9◄;
(i)brucellosis-free bovine herd means a bovine herd which satisfies the condi-
tions laid down in ►M1Annex A.II, paragraphs 4 and 5 ◄;
(j)officiallyenzootic-bovine-leukosis-free herd means a herd which satisfies
the conditions laid down in Annex D, Chapter I, Sections A and B;
(k)officiallyenzootic-bovine-leukosis-free Member State orregionmeans a
region or Member State which meets the requirements laid down in
►M1Annex D, Chapter I, Sections E and F ◄;
(l)official veterinarian means the veterinarian appointed by the competent
authority of the Member State;1997L0012 — EN — 31.12.1998 — 001.001 — 4
(1) OJ No L 47, 21. 2. 1980, p. 11. Directive as last amended by Directive 93/384/EEC
(OJ No L 166, 8. 7. 1993, p. 34).
(2) OJ No L 47, 21. 2. 1980, p. 4. Directive as last amended by Directive 91/687/EEC (OJ
No L 377, 31. 12. 1991, p. 16).
(3) OJ No L 194, 22. 7. 1988, p. 10. Directive as last amended by Directive 93/60/EEC
(OJ No L 186, 28. 7. 1993, p. 28).
(4) OJ No L 351, 2. 12. 1989, p. 34.
(5) OJ No L 224, 18. 8. 1990, p. 62. Directive as last amended by the 1994 Act of Acces-
sion.
(6) OJ No L 363, 27. 12. 1990, p. 51. Directive as last amended by Directive 92/118/EEC.
(7) OJ No L 340, 11. 12. 1991, p. 17. Directive as last amended by Directive 95/29/EC (OJ
No L 148, 30. 6. 1995, p. 52).
(8) OJ No L 355, 5. 12. 1992, p. 32. Directive as last amended by the 1994 Act of Acces-
sion.
(9) OJ No L 224, 18. 8. 1990, p. 19. Decision as last amended by Decision 94/370/EC (OJ
No L 168, 2. 7. 1994, p. 31).
▼B
(m)approved veterinarian means any veterinarian approved by the competent
authority in accordance with the provisions of Article 14 (3) (B);
(n)compulsorilynotifiable diseases means the diseases listed in Annex E (I);
(o)assemblycentre means holdings, collection centres and markets, at which
bovine animals or swine originating from different holdings are grouped
together to form consignments of animals intended for trade. These
assembly centres must be approved for trading purposes and meet the
requirements laid down in Article 11;
(p)regionmeans that part of a Member State's territory which is at least
2 000 km2in area and which is subject to inspection by the competent
authorities and includes at least one of the following administrative regions:
— Belgium: province — provincie
— Germany: Regierungsbezirk— Denmark: amt or island
— France: département
— Italy: provincia— Luxemburg —
— Netherlands: rrv-kring
— United Kingdom: England, Wales and Northern Ireland:
county
Scotland: district or island area
— Ireland: county
— Greece: νοµός
— Spain: provincia
— Portugal: continente: distrito, and other parts of
Portugal's territory: região autónoma
— Austria: Bezirk
— Sweden: län
— Finland: lääni/län;
(q)dealermeans any natural or legal person who buys and sells animals
commercially either directly or indirectly, who has a regular turnover of
these animals and who within a maximum of 30 days of purchasing animals
resells them or relocates them from the first premises to other premises not
within his ownership and meets the conditions laid down in Article 13.
Article 3
1. Each Member State shall ensure that only animals that fulfil the relevant
conditions laid down in this Directive are sent from its territory to that of
another Member State.
2. Bovine animals and swine covered by this Directive must:
(a) be subjected:
— to an identity check, and
— to a clinical inspection within 24 hours of departure by an official veter-
inarian and show no clinical sign of disease;
(b) not have been obtained from a holding or an area which for health reasons
is subject to prohibition or restriction affecting the species involved in
accordance withCommunity and/or national legislation;
(c) be identified as provided for in Directive 92/102/EEC;(d) not be animals which are to be slaughtered and not be restricted under a
contagious or infectious disease eradication programme of a Member State
or region;
(e) comply withth e provisions of Articles 4 and 5.
Article 4
1. Bovine animals and swine covered by this Directive must at no time
between leaving the holding of origin and arriving at destination come into1997L0012 — EN — 31.12.1998 — 001.001 — 5
▼B
contact with cloven-hoofed animals other than animals that have the same health
status.
2. Bovine animals and swine covered by this Directive must be transported in
means of transport meeting the requirements of Directive 91/628/EEC and in
addition the requirements of Article 12.
3. Rules for the approval of sites where cleansing and disinfection may be
carried out shall be determined in accordance with the procedure set out in
Article 17.
Article 5
1. Bovine animals and swine covered by this Directive must be accompanied
during transportation to destination by a health ►M1certificate conforming to
either model 1 or 2 set out in Annex F as appropriate ◄. The certificate shall
consist of a single sheet or, where more than one page is required, shall be in
such a form that any two or more pages are part of an integrated whole and
indivisible and shall contain a serial number. It shall be drawn up on the day
of the health inspection, in one of the official languages of the country of desti-
nation at least. The certificate shall be valid for 10 days from the date of the
health examination.
2. The health inspection for the issuing of the health certificate (including
additional guarantees) for a consignment of animals may be carried out in the
holding of origin or of an assembly centre. For this purpose the competent
authority shall ensure that any health certificate is drawn up by the official
veterinarian after inspections, visits and controls as provided by this Directive.
However as regards:
(a) animals coming from approved assembly centres, suchcertification sh all be:
— on the basis of an official document containing the necessary informa-
tion completed by the official veterinarian for the holding of origin, or
— in the form of the ►M1certificate according to either model 1 or 2 in
Annex F as appropriate ◄withSections A and B duly completed and
certified by the official veterinarian for the holding of origin;
(b) animals coming from an approved holding which is participating in the
surveillance network provided for in Article 14, suchcertification sh all be:
— on the basis of an official document containing the necessary informa-
tion completed by the approved veterinarian for the holding of origin, or
— in the form of the ►M1certificate according to either model 1 or 2 in
Annex F as appropriate ◄withSections A and B duly completed and
certified by the approved veterinarian for the holding of origin.
For this purpose, the official veterinarian will ensure where appropriate that the
additional guarantees provided for in Community legislation are fulfilled.
3. The official veterinarian for the assembly centre shall carry out all neces-
sary checks on animals arriving there.
4. The official veterinarian completing ►M1Section C of the certificate
according to either model 1 or 2 in Annex F as appropriate ◄, shall ensure
that the movement is registered on the Animo system on the day the certificate
is issued.
5. The animals covered by this Directive may transit through an assembly
centre which is located in one other Member State before being consigned to
the Member State of destination. In this case, the ►M1certificate according
to either model 1 or 2 in Annex F as appropriate ◄►M1(including Section
C)◄must be completed by the official veterinarian responsible in the Member
State where the animals originate. The official veterinarian responsible for the
assembly centre of transit shall provide certification to the Member State of
destination by completing a second certificate, as in Annex F, endorsing it with
the serial number of the original and attaching it to the original certificate or to
an officially endorsed copy thereof. In this case the combined validity of the
certificates shall not exceed that provided for in paragraph 1.
Article 6
1. Animals for breeding or production must, in addition to the requirements
of Articles 3, 4 and 5:
— have remained in a single holding of origin for a period of 30 days prior to
loading, or since birth in the holding of origin where the animals are less
than 30 days old. The official veterinarian must, on the basis of the official
identification provided for in Article 3 (2) (c) and official records, be satis-
fied that the animals have complied with this condition and furthermore that1997L0012 — EN — 31.12.1998 — 001.001 — 6
▼B
the animals have originated in the Community or have been imported from a
third country in compliance with Community animal health legislation.
However, in the case of animals transiting through an approved assembly
centre in the Member State of origin, the period during which the assembly
of these animals takes place outside the holding of origin shall not exceed
six days,
— withregard to animals imported from a th ird country into a Member State
which is not that of ultimate destination, be transported to the Member State
of destination as quickly as practicable under cover of a certificate issued
under Article 7 of Directive 91/496/EEC,
— withregard to animals imported from a th ird country upon arrival at destina-
tion and before any further movements satisfy the requirements of this
Directive, and in particular the residency requirement in the first indent,
and may not be brought into the herd until the veterinarian responsible for
that holding has ascertained that the animals in question are not likely to
jeopardize the health status of the holding.
If an animal from a third country is introduced into a holding no animal
from the holding may be traded for 30 days following introduction unless
the imported animal is isolated from all other animals on the holding.
2. Bovine animals for breeding and production must, in addition to the
requirements in Articles 3, 4 and 5:
(a) come frome an officially tuberculosis-free bovine herd, and in the case of
animals more than six weeks old, have reacted negatively to an intradermal
tuberculin test carried out during the 30 days prior to leaving the herd of
origin, in accordance withth e provisions of Annex B point 32 (d).
This intradermal tuberculin test is not required if the animals originate in a
Member State or part of a Member State recognized as officially tubercu-
losis free or in a Member State or part of a Member State withan
approved surveillance network;
(b) in the case of uncastrated animals which come from an officially brucel-
losis-free bovine herd and more than 12 months old, have shown a brucella
count lower that 30 international units (IU) of agglutination per millilitre
when given a serum agglutination test (or any test approved by Standing
Veterinary Committee (SVC) procedure following the adoption of the rele-
vant protocols) carried out during the 30 days prior the leaving the herd
oforigin and complying withth e provisions of Annex C Section A.
This serum agglutination test (or any test approved by SVC procedure
following the adoption of the relevant protocols) is not required if the
animals originate in a Member State or part of a Member State recognized
as officially brucellosis free or in a Member State or part of a Member State
withan approved surveillance network;
(c) come from an officially enzootic-bovine-leukosis-free herd and, if more than
12 months old, have reacted negatively to an individual test carried out
during the 30 days prior to leaving the herd of origin and complying with
the provisions of Annex D.
The test is not required if the animals originate in a Member State or part of
a Member State recognized as officially enzootic-bovine-leukosis free or in
a Member State or part of a Member State withan approved surveillance
network;
(d) at no time between leaving the holding of origin and arriving at destination
come into contact with bovine animals which meet only the requirements in
paragraph3.
3. Bovine animals for slaughter must, in addition to the requirements in Arti-
cles 3, 4 and 5, come from herds that are officially tuberculosis free, officially
enzootic-bovine-leukosis free and in the case of uncastrated bovines, from herds
that are officially brucellosis free.
However, until 31 December 1999, the destination countries may grant to Spain
general or limited licences to introduce into their territories animals for
slaughter from herds which are not officially free of tuberculosis, enzootic
bovine leukosis and brucellosis, provided suchanimals:
— have in the 30 days prior to embarkation undergone the appropriate tests laid
down in Annexes B, C and D, withnegative results,
— are taken on arrival in the country of destination directly to a slaughterhouse
and are slaughtered there as soon as possible but at least within 72 hours of
arrival, in accordance withanimal h ealthrequirements.1997L0012 — EN — 31.12.1998 — 001.001 — 7
▼B
Article 7
Animals for slaughter which have been taken on arrival in the country of desti-
nation:
— to a slaughterhouse, must be slaughtered there as soon as possible but at
least within 72 hours of arrival, in accordance with animal health require-
ments, or
— to an approved assembly centre, must be removed after the market directly
to a slaughterhouse to be slaughtered as soon as possible but at the latest
within three working days of arrival at the assembly centre, in accordance
with animal health requirements. At no time, between their arrival at the
assembly centre and their arrival at the slaughterhouse, may they come into
contact with cloven-hoofed animals other than animals that fulfil the condi-
tions laid down in this Directive.
Article 8
Member States shall ensure that the suspected presence of any of the diseases
referred to in Annex E (I) is compulsorily and immediately notifiable to the
competent authority.
Each Member State shall forward to the Commission by 31 May each year, and
for the first time in 1999, details of the occurence of diseases listed in Annex E
(I) and of any other diseases covered by the additional guarantees provided for
by Community legislation in its territory in the previous calendar year including
details of the monitoring and eradication programmes in operation. This infor-
mation shall be based on uniform criteria to be established by the procedure
provided for in Article 17. The Commission shall present this information to
the Member States in the framework of the SVC and in particular may utilize
it in relation to the decisions referred to in Annexes A and D.
Article 9
1. A Member State which has a compulsory national control programme for
one of the contagious diseases listed in Annex E (II) for all or part of its terri-
tory may submit the said programme to the Commission, outlining in particular:
— the distribution of the disease in the Member State,
— the reasons for the programme, taking into consideration the importance of
the disease and the programme's likely benefit in relation to its cost,
— the geographical area in which the programme will be implemented,
— the status categories to be applied to the animal establishments, the standards
which must be attained in each category, and the test procedures to be used,
— the programme monitoring procedures, the results of which must be supplied
at least annually to the Commission,
— the action to be taken if, for any reason, an establishment loses its status,
— the measures to be taken if the results of the tests carried out in accordance
with the provisions of the programme are positive.
2. The Commission shall examine the programmes presented by the Member
States. Programmes as referred to in paragraph1 may be approved in compli-
ance with the criteria laid down in paragraph 1 in accordance with the
procedure provided for in Article 17. According to the same procedure, the
additional guarantees, general or limited, which may be required in intra-
Community trade, shall be defined at the same time or at the latest three months
after approval of the programmes. Such guarantees must not exceed those which
the Member State implements nationally.
3. Programmes submitted by Member States may be amended or supple-
mented in accordance withth e procedure laid down in Article 17.
Amendments or additions to programmes which have already been approved or
to guarantees which have been defined in accordance with paragraph 2 may be
approved under the same procedure.
Article 10
1. Where a Member State considers that its territory or part of its territory is
free from one of the diseases listed in Annex E (II), it shall present to the
Commission appropriate supporting documentation, setting out in particular:
— the nature of the disease and the history of its occurrence in its territory,
— the results of surveillance testing based on serological, microbiological,
pathological or epidemiological investigation and on the fact that the disease
must by law be notified to the competent authorities,
— the period over which the surveillance was carried out,
— where applicable, the period during which vaccination against the disease
has been prohibited and the geographical area concerned by the prohibition,1997L0012 — EN — 31.12.1998 — 001.001 — 8
▼B
— the arrangements for verifying the absence of the disease.
2. The Commission shall examine documentation submitted by Member
States. The additional guarantees, general or specific, which may be required
in intra-Community trade may be defined in accordance withth e procedure
laid down in Article 17. Such guarantees must not exceed those which the
Member State implements nationally.
3. The Member State concerned shall notify the Commission of any change
in the details specified in paragraph 1 which relate to the disease, in particular
regarding any new outbreaks of the disease. The guarantees defined in accor-
dance with paragraph 2 may, in the light of such notification, be amended or
withdrawn in accordance with the procedure laid down in Article 17.
Article 11
1. Member States shall ensure that, in order to be approved by the competent
authority, assembly centres meet the following conditions at least. They must:
(a) be under the control of an official veterinarian who shall ensure that, in
particular, the provisions of Article 4 (1) and (2) are complied with;
(b) be located in an area which is not subject to prohibiiton or restrictions in
accordance withrelevant Community legislation and/or national legislation;
(c) be cleaned and disinfected before use, as required by the official veteri-
narian;
(d) they must have, taking into account the animal capacity of the assembly
centre:
— a facility dedicated exclusively for this purpose when used as an
assembly centre,
— appropriate facilities for loading, unloading and adequate housing of a
suitable standard for the animals, for watering and feeding them, and
for giving them any necessary treatment; these facilities must be easy
to clean and disinfect,
— appropriate inspection facilities,
— appropriate isolation facilities,
— appropriate equipment for cleaning and disinfecting rooms and trucks,— an appropriate storage area for fodder, litter and manure,— an appropriate system for collecting waste water,— the use of an office for the official veterinarian;
(e) admit only animals that are identified and come from herds that are offi-
cially free of tuberculosis, brucellosis and leukosis or slaughter animals
meeting the conditions set out in the present Directive and in particular
Article 6 (3). To this end, when animals are admitted the owner or person
in charge of the centre shall ensure they are properly identified and accom-
panied by health documents or appropriate certificates for the species and
categories involved;
(f) be regularly inspected in order to ascertain that the requirements for
approval continue to be fulfilled.
2. The owner or person in charge of the assembly centre shall be required, on
the basis either of theaccompanying documents for the animals or of the identi-
fication numbers or marks of the animals, to record on a register or a data base
and retain for a minimum period of three years the following information:
— the name of the owner, the origin, date of entry and exit, number and iden-
tification of the bovine animals or the registration number of the holding of
origin or of the herd of origin of the pigs entering the centre and their
proposed destination,
— the registration number of the transporter and the licence number of the lorry
delivering or collecting animals from the centre.
3. The competent authority shall issue an approval number to each approved
assembly centre. Suchapproval may be limited to a particular species or to
animals for breeding and production or to animals for slaughter. The competent
authority shall notify the Commission of the list of approved assembly centres
and of any updates. The Commission shall present this information to Member
States in the framework of the SVC.
4. The competent authority may suspend or withdraw approval in the event
of failure to comply with this Article or other appropriate provisions of this
Directive or other directives in respect of health restrictions. Approval may be
restored when the competent authority is satisfied that that assembly centre is in
full compliance with all the appropriate provisions of this Directive.
5. The competent authority shall ensure that when operating assembly centres
have sufficient approved veterinarians to carry out all duties.1997L0012 — EN — 31.12.1998 — 001.001 — 9
▼B
6. Any detailed rules required for uniform application of this Article shall be
adopted in accordance withth e procedure provided for in Article 17.
Article 12
1. Member States shall ensure that the transporters referred to in Article 5 of
Directive 91/628/EEC meet the following additional conditions:
(a) for the carriage of animals they must use means of transport that are:
— constructed in such a way that the animal faeces, litter or feed cannot
leak or fall out of the vehicle,
— cleaned and disinfected immediately after every animal transport opera-
tion or that of any product which could affect animal health and if
necessary before any new loading of animals, using disinfectants offi-
cially authorized by the competent authority;
(b) they must either have appropriate cleaning and disinfection facilities
approved by the competent authority, including facilities for storing litter
and dung, or they must provide documentary evidence that these operations
are perfomed by a third party approved by the competent authority.
2. The transporter must ensure that for each vehicle used for the transport of
animals a register is kept containing the following information at least which
shall be retained for a minimum period of three years:
(i) places and dates of pick-up, and the name or business name and address of
the holding or assembly centre where the animals are picked up;
(ii) places and dates of delivery, and the name or business name and address of
the consignee(s);
(iii) species and number of animals carried;
(iv) date and place of disinfection;
(v) details of accompanying documentation (serial number, etc.).
3. Transporters shall ensure that the consignment of animals do not at any
time, between leaving the holdings or the assembly centre of origin and arriving
at their destination, come into contact with animals of a lower health status.
4. Member States shall ensure that transporters give a written undertaking
stating in particular that:
— all the measures necessary to comply with this Directive shall be taken and
in particular the provisions laid down in this Article and relating to the
appropriate documentation that must accompany the animals,
— the transport of animals is entrusted to staff who possess the necessary
ability, professional competence and knowledge.
5. In the event of failure to comply with the provisions of this Article the
requirements of Article 18 of Directive 91/628/EEC shall apply mutatis
mutandis relating to animal health.
Article 13
1. Member States shall ensure that all dealers are registered, approved and
issued with an approval number by the competent authority and that they
comply withth e following conditions at least:
(a) they must deal only in animals that are identified and come from herds that
are officially free of tuberculosis, brucellosis and leukosis or slaughter
animals meeting the conditions set out in thisDirective and in particular
Article 6 (3). To this end, the dealer shall ensure that the animals are prop-
erly identified and are accompanied by health documents as appropriate forthe species involved.
However, the competent authority may authorize the marketing of identified
animals which do not fulfil the conditions laid down in the first paragraph,
in so far as they are brought direct to a slaughterhouse in the Member State
of origin without passing through their facilities, for slaughter as soon as
possible in order to prevent the spreading of diseases. The necessary provi-
sions should be taken to ensure that such animals, when they reach the
abattoir, cannot come into contact with other animals and that they are
slaughtered apart from other animals;
(b) the dealer shall be required, either on the basis of the document accompa-
nying the animals, or on the basis of identification numbers or marks on the
animals, to keep a record or data base and to store the following data for at
least three years:
— the name of the owner, origin, date of purchase, categories, number and
identification of bovine animals or registration number of the holding of
origin or of the herd of origin of pigs purchased,1997L0012 — EN — 31.12.1998 — 001.001 — 10
▼B
— the registration number of the transporter and/or the licence number of
the lorry delivering and collecting animals,
— the name and address of the purchaser and the destination of the
animals,
— copies of route plans and/or serial number of health certificates as
applicable;
(c) when the dealer keeps animals on his premises he shall ensure that:
— specific training is given to the staff in charge of the animals in applying
the requirements of this Directive and in the care and welfare of the
animals,
— controls and tests if necessary on the animals are carried out regularly
by the official veterinarian and that all necessary steps are taken to
prevent the spread of disease.
2. Member States shall ensure that all premises used by a dealer in connec-
tion with his business are registered and issued with an approval number by the
competent authority and that they comply with the following conditions at least:
(a) they must be under the control of an official veterinarian;
(b) they must be located in an area which is not subject to prohibition or restric-
tions in accordance withth e relevant Community legislation or national
legislation;
(c) they must have:
— appropriate facilities of sufficient capacity and in particular inspection
facilities and isolation facilities so that all animals can be isolated in
the event of an outbreak of a contagious disease,
— appropriate facilities for unloading and where necessary adequate
housing of a suitable standard for the animals, for watering and feeding
them, and for giving them any necessary treatment; these facilities must
be easy to clean and disinfect,
— an appropriate reception area for litter and manure,
— an appropriate system for collecting waste water;
(d) be cleaned and disinfected before use, as required by the official veteri-
narian.
3. The competent authority may suspend or withdraw approval in the event
of failure to comply with this Article or other appropriate provisions of this
Directive or other directives in respect of health restrictions. Approval may be
restored when the competent authority is satisfied that the dealer is in full
compliance with all the appropriate provisions of this Directive.
4. The competent authority must carry out regular inspections in order to
ascertain that the requirements of this Article are fulfilled.
Article 14
1. The competent authority in a Member State may introduce a system of
surveillance networks.
The surveillance network system must comprise at least the following elements:
— the herds,
— the owner or any other natural or legal person responsible for the holding,
— the approved veterinarian or the official veterinarian responsible for the
holding,
— the official veterinary service of the Member State,
— the official veterinary diagnostic laboratories or any other laboratory
approved by the competent authority,
— a computer database.
Official veterinarians for the slaughtering establishments and approved assembly
centres will be associated withth e network system.
2. The main objectives of the surveillance network system are to make the
official classification of holdings, to maintain such classification by regular
inspection, to collect epidemiological data and to carry out disease monitoring
so as to ensure compliance with all the provisions of this Directive and other
directives in respect of health restrictions.
This surveillance network system shall be mandatory on all holdings in the terri-
tory of the Member State operating such a system. However, the competent
authority may authorize the establishment of such a network on part of the terri-
tory made up of one or several adjacent regions as defined in Article 2 (2) (p).
Where this derogation is accorded, animal movements to that part of the terri-1997L0012 — EN — 31.12.1998 — 001.001 — 11
▼B
tory from other regions which are not part of the network system shall be
subject to the provisions of this Directive.
The competent authority shall lay down the obligations and rights incumbent
upon the approved veterinarians, the persons responsible for the holdings or
their owners and any other participants in the system including the persons
responsible for issuing health certificates.
3. The competent authority shall ensure that the obligations referred to in
paragraph2 at least include th e following:
A. Every owner of or person responsible for a holding must:
(i) secure, by contract or legal instrument, the services of a veterinarian
approved by the competent authority;
(ii) immediately call in the approved veterinarian for the holding when he
suspects the occurrence of an infectious disease or of any notifiable
disease;
(iii) notify the approved veterinarian of all arrivals of animals on his
holding;
(iv) isolate the animals before introducing them into his holding to enable
the approved veterinarian to check, where appropriate by means of the
required tests, whether the holding's status may be maintained.
B. The approved veterinarians as provided for by Article 2 (2) (m) shall be
under the control of the competent authority and must comply with the
following requirements.
They must:
(i) meet the conditions for pursuing the veterinary profession;
(ii) have no financial interest or family links with the owner of or person
responsible for the holding;
(iii) possess particular knowledge in the field of animal health as it applies
to animals of the species concerned. This means that they must:
— regularly update their knowledge, especially as regards the relevant
health regulations,
— meet the requirements laid down by the competent authority to
ensure the proper functioning of the network,
— provide the owner of or person responsible for the holding with
information and assistance in order that all steps are taken to ensure
that the holding's status is maintained, particularly on the basis of
programmes agreed with the competent authority,
— ensure compliance withth e requirements concerning:
(i) the identification and health certification of the animals of the
herd, the animals introduced and those traded;
(ii) compulsory reporting of infectious animal diseases and any
other risk factor for animal health or welfare, and for human
health;
(iii) establishing as far as possible the cause of death of animals and
where they are to be consigned;
(iv) the hygiene conditions of the herd and of the livestock produc-
tion units.
If the proper functioning of the system so requires, each Member
State may limit the veterinarians' responsibility to a specific number
of holdings or to a specific geographical area.
The competent authority shall draw up lists of approved veterinar-
ians and of the approved holdings participating in the network. If
the competent authority finds that a participant in the network no
longer fulfils the conditions set out above, it shall suspend or with-
draw approval, without prejudice to any penalties that may be
applied.
C. The computer database must contain at least the following information:
(1) For eachanimal:
— identification code,
— date of birth,— sex,— breed or colour of coat,
— identification code of the mother or, in the case of an animal
imported from a third country, the identification number given
following inspection under Directive 92/102/EEC and corresponding
to the identification number of origin,
— identification number of the holding where born,1997L0012 — EN — 31.12.1998 — 001.001 — 12
▼B
— identification numbers of all holdings where the animal has been
kept and the dates of each change of holding,
— date of deathor slaugh ter.
(2) For eachh olding:
— an identification number consisting of not more than 12 figures
(apart from the country code),
— name and address of the holder.
(3) The database must be able to supply the following particulars at any
time:
— the identification number of all animals of the bovine species present
on a holding, or in the case of groups of animals of the porcine
species, the registration number of the holding of origin or herd of
origin and the number of the health certificate where applicable,
— a list of all changes of holding for each animal of the bovine species
starting from the holding of birth, or the holding of importation in
the case of animals imported from third countries; and for groups of
pigs the registration number of the last holding or last herd and for
imported animals from third countries the holding of importation.
These particulars will be held on the database until three consecutive
years have elapsed since the death of the bovine animal or until three
consecutive years have elapsed since the record was made in the case
of records for pigs.
However, only paragraphs 2 and 3 shall be applicable to pigs.
4. All participants in the surveillance network other than those provided for
in 3A and B shall be accountable to the competent authority. The competent
authority in each Member State shall be responsible for setting up the network
and shall carry out regular checks to ensure that it operates properly.
5. Member States which introduce a system of surveillance networks as
outlined in paragraphs 1 to 4, operational for a period of at least 12 months,
shall apply to the Commission to have it approved under the procedure provided
for in Article 17.
For this purpose the Commission shall examine documentation submitted by
Member States.
The Commission experts shall validate the systems by means of a system of
audits. Where the result of the audit is favourable the Commission shall within
90 days of receipt of the request for approval make a report to the SVC together
withappropriate proposals.
Where repeated offences are noted, the approval of the surveillance network
system can be suspended according to the procedure laid down in Article 17,
at the request of the Commission or of one or more Member States.
6. Member States which have implemented in all their territory a recognized
surveillance network system as laid down in this Article shall be authorized not
to apply the provision referred to in Article 3 (2) (a), second indent to animal
movements referred to by this Directive within their own territory.
7. Not later than 31 December 1999, acting on the basis of a report from the
Commission, accompanied by proposals on which it shall act by a qualified
majority, the Council shall review the provisions of this Article in the light of
experience witha view to amending and updating th em and, if appropriate,
extending them to all Member States.
8. The financing of the surveillance network system will be covered within
the framework of the revision of Annex B to Directive 85/73/EEC(1) in accor-
dance withth e provisions laid down in Article 8 of Directive 96/43/EC.
Article 15
1. Member States shall take the appropriate specific measures to penalize any
infringement of this Directive whether by a natural or a legal person.
2. If it is confirmed that the provisions of this Directive are not or have not
been complied with the competent authority of the place in which such a finding
is made shall take all appropriate measures to safeguard animal health and to
prevent the spread of disease.1997L0012 — EN — 31.12.1998 — 001.001 — 13
(1) OJ No L 32, 5. 2. 1985, p. 14. Directive as amended by Directive 96/43/EC (OJ No L
162, 1. 7. 1996, p. 1).
▼B
Depending on the circumstances, such action by the competent authority may
consist of taking the measures required to:
(a) arrange for the journey to be completed or the animals returned to their
place of departure by the most direct route, providing this course of action
would not further jeopardize the health or welfare of the animals;
(b) arrange for the animals to be held in suitable accommodation with appro-
priate care in the event of interruption of the journey;
(c) arrange for the slaughter of the animals. The destination and use of such
animals after slaughter shall be regulated:
— in accordance withth e provisions of Directive 64/433/EEC(1), or
— in accordance with the provisions of Directive 90/667/EEC where the
health status of the animals cannot be established or where they are
liable to be a risk to animal health or public health. However, where
the provisions of Directive 90/667/EEC should apply, a regularization
period may be accorded to the owner or his agent before that final possi-
bility is invoked. In that case, the provisions of paragraph 3 of this
Article shall apply.
3. The competent authority of the Member State of destination shall immedi-
ately notify the competent authority of the Member State of origin on the
establishment of any infringement of this Directive.
In accordance with the provisions established by Directive 89/608/EEC,
Member States shall provide mutual assistance to one another in the application
of this Directive in order to ensure, in particular, compliance with the provisions
laid down in this Article.
4. This Article shall not affect national rules applicable to penal sanctions.
Article 16
1. Annexes A, D (Chapter I), E and F shall be amended by the Council,
acting by a qualified majority on a Commission proposal, in particular with
regard to their adaptation to technological and scientific developments.
Annexes B, C and D (Chapter II) shall be amended by the Commission in accor-
dance withth e procedures provided for in Article 17.
However,
(a) before 1 July 1997, the Commission shall submit to the Council a proposal
for the amendment of Annexes A and D (Chapter I), for the purpose of
updating them and if necessary it will apply the same procedure to Annex
F. The Council shall decide on these proposals by a qualified majority
before 1 January 1998;
(b) before 30 June 1998, the Commission, in accordance with the procedure laid
down in Article 17 and on the basis of the opinion of the Scientific Veter-
inary Committee, shall update and if necessary amend Annexes B, C and D
(Chapter II) to adapt them to scientific developments.
2. Not later than 31 December 1999, acting on the basis of a report from the
Commission, accompanied by suitable proposals on which it shall act by a
qualified majority, the Council shall review the provisions of this Directive in
the light of experience with a view to amending and updating them so that
they comply with the rules laid down for completion of the internal market.
Article 17
Where reference is made to the procedure provided for in this Article, the SVC
set up by Decision 68/361/EEC(2) shall act in accordance with the rules laid
down in Article 18 of Directive 89/662/EEC(3).
Article 18
Those Member States which have not introduced an approved surveillance
network system shall ensure that a computer database complying with the provi-
sions laid down in Article 14 is fully operational from 31 December 1999.1997L0012 — EN — 31.12.1998 — 001.001 — 14
(1) OJ No 121, 29. 7. 1964, p. 2012/64. Directive as last amended by Directive 95/23/EC
(OJ No L 243, 11. 10. 1995, p. 7).
(2) OJ No L 255, 18. 10. 1968, p. 23.
(3) OJ No L 395, 30. 12. 1989, p. 13. Directive as last-amended by Directive 92/118/EEC
(OJ No L 62, 15. 3. 1993, p. 49).
▼B
Article 19
The rules laid down in Directive 90/425/EEC shall apply in particular to checks
at origin, to the organization of, and follow-up to, the checks to be carried out
by the country of destination, and to the safeguard measures to be implemented.
Article 20
This Directive is addressed to the Member States.1997L0012 — EN — 31.12.1998 — 001.001 — 15
▼B
ANNEX A
I.Officially tuberculosis-freebovineherd
1. A bovine herd is officially tuberculosis free if:
(a) all the animals are free from clinical signs of tuberculosis;
(b) all the animals over six weeks old have reacted negatively to at least
two official intradermal tuberculin tests carried out in accordance
with Annex B, the first six months after the elimination of any infec-
tion from the herd and the second six months later or, where the herd
has been assembled solely from animals that originate in officially
tuberculosis-free herds, the first test shall be carried out at least 60
days after assembly and the second shall not be required;
(c) following the completion of the first test referred to in (b), no bovine
animal over six weeks old has been introduced into the herd unless it
has reacted negatively to an intradermal tuberculin test performed and
assessed according to Annex B and carried out either in the 30 days
prior to, or the 30 days after the date of its introduction into the herd.
This test shall not be necessary in Member States or regions of a
Member State where the percentage of bovine herds infected with
tuberculosis is less than 0,2 % or if the animal originates in an offi-
cially tuberculosis-free herd.
2. A bovine herd will retain officially tuberculosis-free status if:
(a) the conditions detailed in 1 (a) and (c) continue to apply;
(b) all animals entering the holding come from herds of officially tuber-
culosis-free status;
(c) all animals in the holding, with the exception of calves under six
weeks old which were born in the holding, are subjected to routine
tuberculin testing in accordance withAnnex B at yearly intervals.
However, the Commission, in accordance with the procedure in
Article 17, may, for a Member State or part of a Member State where
all the bovine herds are subject to official operations to combat tuber-
culosis, alter the frequency of the routine tests as follows:
— if the percentage of bovine herds infected with tuberculosis is not
more than 1 % on average during the two most recent annual
supervisory periods, the interval between routine herd tests may
be increased to two years,
— if the percentage of infected bovine herds is not more than 0,2 %
on average during the two most recent biennial supervisory
periods, the interval between routine tests may be increased to
three years,
— if the percentage of infected bovine herds is not more than 0,1 %
on average during the two most recent triennial supervisory
periods, the interval between routine tests may be increased to
four years and/or the age at which animals have to undergo these
tests may be increased to 24 months.
The Commission may also, in accordance with Article 17, take a
decision increasing the frequency of routine tuberculin testing if the
level of disease appears to have increased.
When a Member State operates a system of identification and regis-
tration of bovine animals which provides for identification of herds of
origin and transit, if the percentage of infected herds is not greater
than 0,1 % on average during the two most recent control periods, it
may dispense with the annual tuberculin test of the herds on condition
that:
(1) before the introduction into the herd all the bovine animals are
subjected to an intradermal tuberculin test withnegative results;
(2) all bovine animals slaughtered are examined for lesions of tuber-
culosis and any suchlesions are submitted to a bacteriological
examination for evidence of tuberculosis bacteria.
3. The officially tuberculosis-free status of a herd shall be suspended if:
(a) the conditions detailed in 2 have not been complied with;
(b) an animal is deemed to have reacted positively to a routine tuberculin
test, or a case of tuberculosis has been diagnosed at routine post-
mortem examination.
In these cases, the status shall remain suspended until such time as all
the remaining animals over six weeks of age have reacted negatively1997L0012 — EN — 31.12.1998 — 001.001 — 16
▼B
to at least two official intradermal tuberculin tests in accordance with
Annex B, the first one carried out at least two months after elimina-
tion of the animal from the holding and the second one at least 42
days after the first.
However, by way of derogation, if during a routine herd test one or
more animals react positively to a tuberculin test, and bovine tubercu-
losis is not considered to be the cause of this reaction, the suspected
case(s) of tuberculosis shall be fully investigated, including in parti-
cular the tracing and checking of the herd of residence at the time
of the test and of any previous herd considered necessary by the
competent authority and all examinations including appropriate post-
mortem tests and laboratory examinations.
While such examinations take place the officially tuberculosis-free
status of the herd referred to shall be suspended until investigations
and laboratory examinations or tuberculin tests have ruled out the
presence of bovine tuberculosis. If the presence of tuberculosis is
not confirmed the status of the herd may be restored.
However, where the routine herd test detailed in 2 (c) has not been
performed on time, the status of the herd shall not be suspended
provided that the test is carried out not later than 60 days after it
was originally due, and provided that subsequent testing takes place
according to the original timetable;
(c) the herd contains animals of unresolved status as described in Annex
B, point 32. In this case the status of the herd shall remain suspended
until the animals' status has been clarified.
4. A Member State or part of a Member State may be declared officially
tuberculosis free according to the procedure laid down in Article 17 if it
meets the following conditions:
(a) the percentage of infected bovine herds has not been more than
0,01 % for six consecutive years and at least 99,9 % of the herds
have been declared officially tuberculosis free for 10 years;
(b) an identification system making it possible to identify the herds of
origin and transit for eachbovine animal is in existence;
(c) all bovine animals slaughtered must be subjected to a post-mortem
examination by an official veterinarian;
(d) all suspected cases of tuberculosis must be fully investigated,
including tracing and checking any herds of origin or transit and
carrying out all appropriate laboratory examinations. While such
examinations take place the officially tuberculosis-free status of the
herds of origin or transit shall be suspended until clinical or labora-
tory examinations or tuberculin tests have ruled out the presence of
bovine tuberculosis.
5. The Member State or part of a Member State will retain officially tuber-
culosis-free status if:
(a) the conditions 4 (a) to (d) continue to apply;
(b) when a case of tuberculosis is confirmed, the officially tuberculosis-
free status of the herd of origin and transit is withdrawn;
(c) the officially tuberculosis-free status of herds where tuberculosis has
been confirmed remains withdrawn until:
— all the animals that have been deemed to be infected have been
slaughtered,
— disinfection of the premises and utensils has taken place,
— all the remaining bovine animals over six weeks of age have
reacted negatively to at least two official intradermal tests in
accordance with Annex B, the first at least six months after the
removal of the infected animals, the second six months after the
first.
6. If there is evidence of a significant change in the situation as regards
tuberculosis in a Member State or part of a Member State which has
been recognized as officially tuberculosis free, the Commission may in
accordance withth e procedure laid down in Article 17 take a decision
suspending or revoking the status and requiring routine tuberculin tests
to be carried out in accordance with one of the schedules in 2 (c).
II.Officially brucellosis-free andbrucellosis-free bovineherds
For the purposes of this Section ‘bovine animals’ means all bovine animals
with the exception of males castrated before the age of four months.1997L0012 — EN — 31.12.1998 — 001.001 — 17
▼B
1. A bovine herd is officially brucellosis free if:
(a) it contains no bovine animals which have been vaccinated against
brucellosis, except females which have been vaccinated at least three
years previously;
(b) all the bovine animals have been free from clinical signs of brucel-
losis for at least six months;
(c) all the bovine animals over 12 months old have been subjected to
one of the following test regimes with negative results in accordance
withAnnex C:
(i) two serum agglutination tests at an interval of more than three
months and less than 12 months;
(ii) three ring tests at three-monthly intervals followed at least six
weeks later by a serum agglutination test;
(iii) two buffered brucella antigen tests at an interval of more than
three months and less than 12 months;
(iv) two micro-agglutination tests at an interval of more than three
months and less than 12 months.
2. A bovine herd will retain officially brucellosis-free status if:
(a) one of the following test regimes is carried out annually with nega-
tive result in accordance withAnnex C:
(i) three ring tests carried out at intervals of at least three months;
(ii) three milk Elisas carried out at intervals of at least three
months;
(iii) two ring tests carried out at an interval of at least three months
followed at least six weeks later by a serological test;
(iv) two milk Elisas carried out at an interval of at least three
months followed at least six weeks later by a serological test;
(v) two serological tests carried out at an interval of at least three
months and not more than six months.
However, the Commission, in accordance with the procedure laid
down in Article 17, may, for a Member State or part of a Member
State which is not officially brucellosis free but where all the bovine
herds are subject to official operations to combat brucellosis, alter
the frequency of the routine tests as follows:
— where not more than 1 % of bovine herds are infected, it may be
sufficient to carry out eachyear two ring tests or two milk Elisas
at an interval of at least three months, or one serological test,
— where at least 99,8 % of bovine herds have been recognized as
officially brucellosis free for at least four years, the interval
between checks may be extended to two years and the checks
must be carried out using one of the serological tests referred to
in' paragraph7 (a);
(b) all bovine animals entering the herd come from herds of officially
brucellosis-free status and, in the case of bovine animals over 12
months old, have shown a brucella count of less than 30 IU of
agglutination per ml when given a serum agglutination test in accor-
dance with Annex C during the 30 days prior to introduction into the
herd.
However, the serum agglutination test described in point (b) need
not be required in Member States, or regions of Member States,
where the percentage of bovine herds infected with brucellosis has
not exceeded 0,2 % for at least two years and where the animal
comes from an official brucellosis-free bovine herd within that
Member State or region and has not during transportation come
into contact withbovine animals of lesser status;
(c) notwithstanding point (b), bovine animals from a brucellosis-free
bovine herd may be introduced into an officially brucellosis-free
herd if they are at least 18 months old and, if vaccinated against
brucellosis, the vaccination was carried out more than a year
previously.
Such animals must have shown, in the 30 days prior to introduction,
a brucella count lower than 30 IU of agglutination per ml and a
negative result when given a complement fixation test, both in
accordance withAnnex C.
If, however, a bovine animal from a brucellosis-free herd is intro-
duced into an officially brucellosis-free bovine herd, under these
provisions, that herd shall be considered to be brucellosis free for
two years from the date on which the animal was introduced.1997L0012 — EN — 31.12.1998 — 001.001 — 18
▼B
3. The officially brucellosis-free status of a herd may be suspended or
withdrawn if:
(a) the conditions detailed in paragraphs 1 and 2 have not been
complied with; or
(b) as a result of laboratory tests or on clinical grounds one or more
bovine animals are suspected of having brucellosis.
If one or more bovine animals in an officially brucellosis-free herd are
suspected of having brucellosis, the status of the herd may be suspended,
rather than withdrawn, if the animal or animals are immediately
destroyed or isolated.
Where the animal has been destroyed, the suspension may be lifted if
two serum agglutination tests, carried out in accordance withAnnex C
on all bovine animals in the herd over 12 months old, show a count
lower than 30 IU of agglutination per ml. The first test shall be carried
out at least 30 days after the removal of the animal and the second at
least 60 days later.
Where the animal has been isolated, it may be reintroduced into the herd
and the status of the herd may be restored, if it subsequently shows a
serum agglutination count lower than 30 IU of agglutination per ml and
has given a negative result to a complement fixation test, these tests
being carried out in accordance withAnnex C.
Where, as a result of laboratory tests or epidemiological investigations,
brucella infection has been confirmed in a herd, the status of that herd
shall not be restored until all bovine animals that were pregnant at the
time of the outbreak have given negative results to the above tests, the
final test having being carried out at least 21 days after calving.
4. A bovine herd is brucellosis free if it complies with the conditions in 1
(a), (b) and (c), except that:
(i) female bovine animals may be vaccinated:
— before the age of six months old with live strain 19 vaccine or
other vaccines approved under the procedure laid down in
Article 17, or
— before the age of 15 months old with killed 45/20 adjuvant
vaccine which has been officially inspected and recognized;
(ii) bovine animals under 30 months old which have been vaccinated
withlive strain 19 vaccine may give a serum agglutination test
result greater than 30 IU but less than 80 IU of agglutination per
millilitre provided that, on the complement fixation test, they give
a result less than 30 EEC units in the case of females vaccinated
less than 12 months previously or less than 20 EEC units in all
other cases;
(iii) in addition to those tests listed in 1 (c), the following test regimes
will also be approved in order to attain brucellosis-free status;
(a) two buffered brucella antigen tests carried out at an interval of
more than three months and less than 12 months;
(b) two micro-agglutination tests carried out at an interval of more
than three months and less than 12 months in accordance with
the provisions of Annex C.
5. A bovine herd will retain brucellosis-free status if:
(i) it is subject to one of the testing regimes listed in 2 (a);
(ii) bovine animals entering the herd comply with the requirements of 2
(b); or
— come from herds of brucellosis-free status, and in the case of
bovine animals over 12 months old, have shown, in the 30 days
prior to introduction into the herd, less than 30 IU of agglutina-
tion per ml when given a serum agglutination test and a negative
complement fixation test in accordance withAnnex C, or
— come from herds of brucellosis-free status, are under 30 months
old and have been vaccinated with live strain 19 vaccine may
give a serum agglutination test result greater than 30 IU but
less than 80 IU of agglutination per millilitre provided that, on
the complement fixation test, they give a result less than 30
EEC units in the case of females vaccinated less than 12 months
previously or less than 20 EEC units in all other cases.
6. The brucellosis-free status of a herd shall be suspended or withdrawn if:
(a) the conditions detailed in paragraphs 4 and 5 have not been
complied with; or1997L0012 — EN — 31.12.1998 — 001.001 — 19
▼B
(b) as a result of laboratory tests or on clinical grounds one or more
bovine animals over 30 months old are suspected of having brucel-
losis.
If one or more bovine animals, over 30 months old, in a brucellosis-free
herd are suspected of having brucellosis, the status of the herd may be
suspended, rather than withdrawn, if the animal or animals are immedi-
ately destroyed or isolated.
Where the animal has been destroyed, the suspension may be lifted if
two serum agglutination tests, carried out in accordance withAnnex C
on all bovine animals in the holding over 12 months old, show a count
lower than 30 IU of agglutination per ml. The first test shall be carried
out at least 30 days after the removal of the animal and the second at
least 60 days later.
Where the animal has been isolated, it may be reintroduced into the herd
and the status of the herd may be restored, if it subsequently shows a
serum agglutination count lower than 30 IU of agglutination per ml and
has given a negative result to a complement fixation test, these tests
being carried out in accordance withAnnex C.
Where, as a result of laboratory tests or epidemiological investigations,
brucella infection has been confirmed in a herd, the status of that herd
shall not be restored until all bovine animals that were pregnant at the
time of the outbreak have given negative results to the above tests, the
final test having being carried out at least 21 days after calving.
7. A region of a Member State may be declared officially brucellosis free
according to the procedure laid down in Article 17 if it meets the
following conditions:
(a) no case of abortion due to brucella infection has been recorded for
at least three years and at least 99,8 % of the herd have been
declared officially brucellosis free for 10 years;
(b) an identification system making it possible to identify the herds of
origin and transit for eachbovine animal is in existence.
8. Subject to point 9, a region declared officially brucellosis free shall
retain this status if all bovine animals over 24 months old are subjected
to either two ring tests or one serological test every three years. In the
event of a positive result the provisions of point 6 shall apply.
9. A region declared officially brucellosis free shall report the occurrence
of all cases of brucellosis to the Commission. The Commission may
according to the procedure laid down in Article 17 propose that the
status be suspended or revoked and require that routine brucellosis
testing be carried out in accordance with one of the schedules in para-
graph2.
10. A Member State may be declared officially brucellosis free according to
the procedure laid down in Article 17 if it meets the following condi-
tions:
(a) no case of abortion due to brucella infection has been recorded for
at least three years and at least 99,8 % of the holdings have been
declared officially brucellosis free for 10 years;
(b) an identification system making it possible to identify the herds of
origin and transit for eachbovine animal is in existence.
11. A Member State declared officially brucellosis free shall retain this
status if:
— every bovine animal suspected of being infected withbrucellosis is
notified to the competent authority and undergoes official investiga-
tion for brucellosis comprising at least two serological blood tests,
including the complement fixation test, and a microbiological exam-
ination of appropriate samples taken in the case of an abortion,
— during the period of suspicion, which shall continue until negative
results have been obtained from the tests provided for in the first
indent, the officially brucellosis-free status of the herd of origin or
transit of the suspected bovine shall be suspended,
— in the event of a positive result, the provisions of point 6 apply.
12. A Member State declared officially brucellosis free shall report the
occurrence of all cases of brucellosis to the Commission. The Commis-
sion may, according to the procedure laid down in Article 17, propose
that the status be suspended or revoked and require that routine brucel-
losis testing be carried out in accordance with one of the schedules in
paragraph2.1997L0012 — EN — 31.12.1998 — 001.001 — 20
▼B
13. (a) For the purposes of Section II, a serological test shall mean either a
serum agglutination test, buffered brucella antigen test, complement
fixation test, plasma agglutination test, plasma ring test, micro-
agglutination test or individual blood Elisa, as described in Annex
C.
(b) Where ring tests are carried out on bulk tanks, the number of those
tests referred to in this Annex shall be doubled and the intervals
between the tests shall be halved.1997L0012 — EN — 31.12.1998 — 001.001 — 21
▼B
ANNEX B
(STANDARDSFOR THEMANUFACTUREANDUSEOF BOVINEAND
AVIANTUBERCULINS)
1. Officially supervised tuberculin tests must be carried out withPPD or
HCSM tuberculins.
2. Manufacturers' working standards for the control of bovine PPD and HCSM
tuberculins must be calibrated in Community tuberculin units (CTU)
following biological assay against the appropriate EEC standard tuberculin.
3. Manufacturers' working standards for the control of avian tuberculins must
be calibrated in international units following biological assay against the
EEC standard for PPD of avian tuberculin.
4. The EEC standard for PPD of bovine tuberculin is that supplied by the
Instituut voor Dierhouderij en Diergezondheid (ID-DLO), Lelystad, The
Netherlands.
5. The EEC standard for bovine HCSM tuberculin is that supplied by the
Institut Pasteur, Paris, France.
6. The EEC standard for avian tuberculin is that supplied by the Veterinary
Laboratory Agency, Addlestone, Weybridge, England.
7. Bovine tuberculins must be prepared withone of th e mycobacterium bovis
strains indicated below:
(a) AN5;
(b) Vallee.
8. Avian tuberculins must be prepared withone of th e mycobacterium avium
strains indicated below:
(a) D4ER;
(b) TB56.
9. The pH of tuberculins must be between 6,5 and 7,5.
10. Anti-microbial preservatives or other substances that may be added to a
tuberculin shall have been shown, to the satisfaction of the State institute
responsible for the official testing of the tuberculin, not to impair the safety
and effectiveness of the product.
The following are the maximum permitted concentrations for phenol and
glycerol:
(a) phenol: 0,5 % m/v;
(b) glycerol: 10 % v/v.
11. Provided the tuberculins are stored at a temperature between 2 and 8 ºC,
protected from light, they may be used up to the end of the following
periods subsequent to the last satisfactory potency test:
(a) liquid PPD tuberculins: two years,
lyophilized PPD tuberculins: eight years;
(b) HCSM tuberculins diluted: two years.
12. The State institutes listed below shall be responsible for the official testing
of tuberculins in their respective countries:
(a) Germany: Paul-Ehrlich Institut, Frankfurt/Main;
(b) Belgium: Instituut Voor Hygiene en Epidemiologie, J. Wytsmanstraat
14, B-1050 Brussels;
(c) France: Laboratoire national des médicaments vétérinaires, Fougères;(d) Grand Duchy of Luxembourg: institute of the supplying country;(e) Italy: Istituto superiore di sanità, Rome;(f) Netherlands: Instituut voor Dierhouderij en Diergezondheid (ID-DLO),
Lelystad;
(g) Denmark: Statens Veterinaere Serumlaboratorium, Copenhagen V;(h) Ireland: institute of the supplying country;(i) United Kingdom: Veterinary Laboratory Agency, Addlestone,
Weybridge;1997L0012 — EN — 31.12.1998 — 001.001 — 22
▼B
(j) Greece: Κέντρο Κτηνιατριχών Ιδρυµάτων Νεαπόλεως 25, 153 10
Αθήνα;
(k) Spain: Laboratorio de Sanidad y producción animal de Granada;(l) Portugal: Laboratorio Nacional de Investigação Veterinária, Lisbon;(m) Austria: Bundesanstalt für Tierseuchenbekämpfung, Mödling;(n) Finland: Eläinlääkintä — ja elintarvikelaitos, Helsinki-anstalten för
veterinärmedicin ochlivsmedel, Helsingfors;
(o) Sweden: Statens veterinärmedicinska anstalt, Uppsala.
13. Official testing must be carried out on eachbatchof bottled tuberculins
ready for use.
14. Tuberculins shall be tested by biological and chemical methods.
15. Tuberculins must be sterile. Tests for sterility shall be carried out according
to the specifications of the European pharmacopoeia.
16. A test for the absence of toxic or irritant properties shall be carried out
according to the specifications of the European pharmacopoeia.
17. Tuberculins must be chemically analyzed to determine the concentration of
glycerol and/or phenol and also the concentration of any other preservative
which may have been added.
18. A test of non-sensitization to tuberculin must be carried out according to the
specifications of the European pharmacopoeia.
19. The potency of tuberculins must be assessed by biological methods. These
methods must be used for HCSM and PPD tuberculins; they are based on
the comparison with standard tuberculins of the tuberculins to be tested.
20. The protein content of PPD tuberculin must be estimated by the Kjeldahl
method. The nitrogen is converted into tuberculo-protein content by multi-
plying by a factor of 6,25.
21. The EEC standard for bovine HCSM has a potency of 65000 Community
tuberculin units (CTU) per ml and is dispensed in ampoules containing 5 ml
of tuberculin.
22. The EEC standard for bovine PPD has a potency of 50000 Community
tuberculin units (CTU) per mg of PPD and is dispensed lyophilized in
ampoules containing 1,8 mg of PPD, i.e. 0,00002 mg PPD has a potency
equal to one Community tuberculin unit.
23. The EEC standard for avian PPD has a potency of 50000 international units
(IU) per mg of the dried material of the purified protein derivative and is
dispensed lyophilized in ampoules containing 10 mg of PPD plus 26,3 mg
of salts, i.e. 0,0000726 mg of the standard has a potency equal to one inter-
national unit.
24. Tuberculins submitted by manufacturers for testing by the State institutes
listed in paragraph12 must h ave been tested for potency by biological assay
against the appropriate standards as listed in paragraphs 2 and 3.
25. (a)Potencytestingonguinea-pigs
Albino guinea-pigs weighing between 400 and 600 g must be used.
These guinea-pigs must be in good health at the time of injection of
the tuberculin. Not less than eight guinea-pigs shall be used for each
assay. The assay should be made not less than one month after sensiti-
zation.
(aa) For the assay of bovine tuberculins, guinea-pigs shall be sensitized
by one of the following methods:
(1) the injection of heat-killed mycobacterium bovis strain AN5 in
oil adjuvant;
(2) the injection of living mycobacterium bovis strain AN5 in
physiological saline;
(3) the injection of BCG vaccine.
(bb) For the assay of avian tuberculins guinea-pigs shall be sensitized
by injection of 2 mg of heat-killed avian-type tubercular bacilli
suspended in 0,5 ml of sterile liquid paraffin or by the injection
of live avian-type tubercular bacilli in physiological saline. The
avian-type strain D4 must be used for this purpose.
(cc) Each tuberculin under test shall be assayed against the appropriate
standard tuberculin by an intradermal assay using groups of
guinea-pigs suitably sensitized.1997L0012 — EN — 31.12.1998 — 001.001 — 23
▼B
The hair shall be clipped from both sides of each guinea-pig. The
assay shall be carried out by comparing the reactions induced by a
series of intracutaneous injections of doses of not more than 0,2 ml
of dilutions of the standard tuberculin in isotonic buffered saline
solution containing Tween 80, 0,0005 %, witha corresponding
series of injections of the tuberculin under test. Dilutions shall be
arranged in geometric series, and injected into guinea-pigs
according to a randomized latin square design (four sites on each
side of an eight-point assay is used). The diameters of the reac-
tions at eachsite sh ould be measured and recorded after 24 to 28
hours.
For eachsample of tuberculin under test, an estimate of relative
potency against the appropriate standard and its fiducial limits
shall be made by statistical methods, using the diameters of the
reactions and the logarithms of the doses as metameters. The
bovine tuberculin under test is of acceptable potency if its esti-
mated potency guarantees per bovine dose 2 000 Community
tuberculin units (more or less 25 %) in cattle. The potency of
eachtuberculin under test sh all be expressed as appropriate in
Community tuberculin units or international units per ml.
(b)Potencytestingoncattle
Periodic potency testing of bovine tuberculins may be carried out on
naturally or artificially infected tuberculous cattle. These potency tests,
on groups of tuberculous cattle, shall be carried out by intradermal four
or six-point assay of the tuberculin under test against the appropriate
standard and the potency of the tuberculin shall be estimated by statis-
tical methods as in the guinea-pig assay.
26. The following requirements shall apply to the labelling of tuberculin
containers and packages.
The label on the containers and the label on the package shall state:
— the name of the preparation,
— for liquid preparations, the total volume in the container,— the number of Community or international units per ml or per mg,— the manufacturer's name,— the batch number,— the nature and quantity of the reconstituting liquid for the freeze-dried
preparation.
The label on the container or the label on the package shall state:
— the expiry date,
— the conditions of storage,— the name and, if possible, the proportions of any added substance,— the strain of bacillus from which the tuberculin has been made.
27. Community laboratories designated in accordance withArticle 17 will be
made responsible for the additional examination of routine issue field tuber-
culins used in the Member States to ensure that the potency of each of these
tuberculins is adequate in relation to the appropriate Community standard
tuberculin. These examinations must be carried out, in tuberculous bovine
animals, in suitably sensitized guinea-pigs and by appropriate chemical
tests.
28. The following shall be recognized as official intradermal tuberculin tests:
(a) the single intradermal test — this test requires a single injection of
bovine tuberculin;
(b) the intradermal comparative test — this test requires one injection of
bovine tuberculin and one injection of avian tuberculin given simulta-
neously.
29. The dose of tuberculin injected shall be:
(1) Not less than 2 000 CTU of bovine tuberculin;
(2) Not less than 2 000 IU of avian tuberculin W15;
The volume of each injection dose shall not exceed 0,2 ml.
30. Tuberculin tests shall be carried out by injecting tuberculin(s) into the skin
of the neck. The injection sites shall be situated at the border of the anterior
and middle thirds of the neck. When both avian and bovine tuberculins are
injected in the same animal, the site for injection of avian tuberculins shall
be about 10 cm from the crest of the neck and the site for the injection of
bovine tuberculin about 12,5 cm lower on a line roughly parallel with the
line of the shoulder or on different sides of the neck; in young animals in1997L0012 — EN — 31.12.1998 — 001.001 — 24
▼B
which there is not room to separate the sites sufficiently on one side of the
neck, one injection shall be made on each side of the neck at identical sites
in the centre of the middle third of the neck.
31. The technique of tuberculin testing and interpretation of reactions shall be
as follows:
(a)Technique
Injection sites shall be clipped and cleansed. A fold of skin within each
clipped area shall be taken between the forefinger and thumb and
measured withcallipers and recorded. A sh ort sterile needle, beveledge outwards, withgraduated syringe ch arged withtuberculin attach ed
shall be inserted obliquely into the deeper layers of the skin. The dose
of tuberculin shall then be injected. A correct injection shall be
confirmed by palpating a small pea-like swelling at eachsite of injec-
tion. The skin-fold thickness of each injection site shall be remeasured
72 hours after injection and recorded.
(b)Interpretation of reactions
The interpretation of reactions shall be based on clinical observations
and the recorded increase(s) in skin-fold thickness at the sites of injec-
tion 72 hours after injection of tuberculin(s).
(ba)Negative reaction: if only limited swelling is observed, withan
increase of not more than 2 mm in the thickness of the fold of
skin without clinical signs such as diffuse or extensive oedema,
exudation, necrosis, pain or inflammation of the lymphatic ductsin that region or of the lymph nodes.
(bb)Inconclusive reaction: if no clinical signs suchas mentioned in
(ba) are observed and if the increase in skin-fold thickness is
more than 2 mm and less than 4 mm.
(bc)Positive reaction: if clinical signs suchas mentioned in (ba) are
observed or there is an increase of 4 mm or more in the thickness
of the fold of skin at the injection site.
32. The interpretation of official intradermal tuberculin tests shall be as follows:
(a) Single intradermal test:
positive: a positive bovine reaction as defined in paragraph31
(bc);
inconclusive: an inconclusive reaction as defined in paragraph31
(bb);
negative: a negative bovine reaction as defined in paragraph31
(ba).
Animals inconclusive to the single intradermal test shall be subjected to
another test after a minimum of 42 days.
Animals which are not negative to this second test shall be deemed to
be positive to the test.
Animals positive to the single intradermal test may be subjected to an
intradermal comparative test.
(b) Intradermal comparative test for the establishment and maintenance of
officially tuberculosis-free herd status:
positive: a positive bovine reaction which is more than 4 mm
greater than the avian reaction, or the presence of clin-
ical signs;
inconclusive: a positive or inconclusive bovine reaction which is
from 1 to 4 mm greater than the avian reaction, and
the absence of clinical signs;
negative: a negative bovine reaction, or a positive or inconclu-
sive bovine reaction but which is equal to or less than
a positive or inconclusive avian reaction and the
absence of clinical signs in bothcases.
Animals inconclusive to the intradermal comparative test shall be
subjected to another test after a minimum of 42 days. Animals which
are not negative to this second test shall be deemed to be positive to
the test.1997L0012 — EN — 31.12.1998 — 001.001 — 25
▼B
(c) Officially tuberculosis-free herd status may be suspended and animals
from the herd shall not be allowed to enter intra-Community trade until
such time as the status of the following animals is resolved:
(1) animals which have been deemed to be inconclusive to the single
intradermal tuberculin test;
(2) animals which have been deemed to be positive to the single intra-
dermal tuberculin test but are awaiting retest withan intradermal
comparative test;
(3) animals which have been deemed to be inconclusive to the intra-
dermal comparative test.
(d) Where animals are required by Community legislation to be subjected
to an intradermal test prior to movement, the test shall be interpreted
so that no animal which shows an increase in skin-fold thickness greater
than 2 mm or the presence of clinical signs is entered into intra-
Community trade.1997L0012 — EN — 31.12.1998 — 001.001 — 26
▼B
ANNEX C
BRUCELLOSIS
A.Serumagglutinationtests
1. The standard agglutinating serum must conform to the standard serum
prepared by the Veterinary Laboratory Agency, Addlestone, Weybridge,
England.
The ampoule must contain 1000 IU of agglutination obtained by lyophi-
lizing 1 ml of bovine serum.
2. The standard serum must be that supplied by the Bundesgesundheitsamt,
Berlin.
3. The degree of brucella agglutination in a serum must be expressed in IU
per ml (i.e. serum X = 80 IU/ml).
4. Readings of slow serum agglutination in tubes must be taken at 50 or at
75 % agglutination, the antigen used having been titrated under identical
conditions against the standard serum.
5. The agglutinating value of various antigens in relation to standard serum
must be within the following limits:
— if the reading is made at 50 %: between 1/600 and 1/1 000,
— if the reading is made at 75 %: between 1/500 and 1/750.
6. Weybridge strain No 99 and USDA 1119 or any other strain of equiva-
lent sensitivity must be used for preparing the antigen for use in tube
agglutination (slow method).
7. The culture media used for keeping the strain in the laboratory and for
producing the antigen must be such that they do not encourage bacterial
dissociation (S-R); potato agar should preferably be used.
8. The bacterial emulsion must be made from physiological saline (NaCl
0,85 ‰ phenolized at 0,5 %. Formol must not be used.
9. The official institutes indicated below must be made responsible for the
official testing of antigens:
(a) Germany: Bundesgesundheitsamt, Berlin;
(b) Belgium: Institut national de recherches vétérinaires, Brussels;(c) France: Laboratoire central de recherches vétérinaires, Alfort;(d) Grand Duchy of Luxembourg: institute of the supplying country;(e) Italy: Istituto superiore di sanità, Rome;(f) Netherlands: Instituut voor Dierhonderij en Diergezondheid (ID-
DLO), Lelystad;
(g) Denmark: Statens Veterinaere Serumlaboratorium, Copenhagen V;(h) Ireland: Veterinary Research Laboratory, Department of Agriculture
and Food, Dublin;
(i) United Kingdom:
— Great Britain: Veterinary Laboratory Agency, Addlestone,
Weybridge, England,
— Northern Ireland: Veterinary Research Laboratory, Stormont,
Belfast;
(j) Greece: Κέντρο Κτηνιατρικών Ιδρυµάτων Νεαπόλεως 25, 153
10 Αθήνα;
(k) Spain: Centro Nacional de Brucelosis; Laboratorio de Sanidad y
Producción Animal de Santa Fé (Granada);
(l) Portugal: Laboratoria Nacional de Investigaçáo Veterinária, Lisbon;(m) Austria: Bundesanstalt für Tierseuchenbekämpfung, Mödling;(n) Finland: Eläinlääkintä-ja elintarvikelaitos, Helsinki-anstalten för
veterinärmedicin ochlivsmedel, Helsingfors;
(o) Sweden: Statens veterinärmedicinska anstalt, Uppsala.
10. Antigens may be delivered in the concentrated state provided the dilu-
tion factor to be used is indicated on the bottle label.
11. In order to carry out a serum agglutination test, at least three dilutions
must be prepared for eachserum. Dilutions of suspect serum must be1997L0012 — EN — 31.12.1998 — 001.001 — 27
▼B
made in such a way that the reading of the reaction at the infection limit
is made in the median tube. If there is a positive reaction in this tube,
the suspect serum contains at least 30 IU of agglutination per ml.
B.Complementfixationreaction test
1. The standard serum is the same as that under A.1 of this Annex. In addi-
tion to its content in international agglutinating units, 1 ml of this
lyophilized bovine serum must contain 1000 sensitizing units which
fix the complement. These sensitizing units are called EEC sensitizing
units.
2. The standard serum must be supplied by the Bundesgesundheitsamt,
Berlin.
3. A serum's level of antibodies which fix the complement must be
expressed in EEC sensitizing units (for example: serum X = 60 EEC
sensitizing units per ml).
4. A serum containing 20 or more EEC sensitizing units (i.e. an activity
equal to 20 % of that of the standard serum) per ml, must be considered
to be positive.
5. Serums must be inactivated as follows:
(a) bovine serum: 56 to 60ºC for 30 to 50 minutes;
(b) swine serum: 60ºC for 30 to 50 minutes.
6. Weybridge strain No 99 or USDA strain 1119 must be used for the
preparation of the antigen. The antigen represents a bacterial suspension
in a physiological serum at 0,85 % or in a veronal loading solution.
7. In order to carry out the reaction test a complementary dose higher than
the minimum necessary for total haemolysis should be used.
8. In carrying out the complement fixation reaction test, the following
controls must be made eachtime:
(a) control of the anti-complementary effect of the serum;
(b) control of the antigen;
(c) control of sensitized red blood corpuscles;
(d) control of the complement;
(e) control using a positive serum of sensitivity at the start of the reac-
tion;
(f) control of the specificity of the reaction using a negative serum.
9. The supervision and official control of standard serums and antigens
shall be carried out by the bodies listed in A.9 of this Annex.
10. Antigens may be delivered in the concentrated state provided the dilu-
tion factor to be used is indicated on the bottle label.
C.Ringtest
1. The ring test must be made on the contents of each milk churn or on the
contents of eachbulk tank from th e farm.
2. The standard antigen to be used must come from one of the institutes
listed in A.9 (a) to (j). It is recommended that the antigens be standar-
dized according to the WHO/FAO recommendations.
3. The antigen may be stained only with haematoxylin or tetrazolium;
haematoxylin should preferably be used.
4. If no preservation is used then the reaction test must be carried out
between 18 and 24 hours of taking the sample from the cow. If milk is
to be tested later than 24 hours after sampling, thenpreservation must be
used; formalin or mercuric chloride may be used as preservatives and if
either of these is used the test must be carried out within 14 days of the
day of sampling. Formalin may be added to give a final concentration in
the milk sample of 0,2 % and, in such cases, the ratio between the
amount of milk and the solution of formalin must be at least 10 to 1. A
solution of mercuric chloride may be used instead of formalin to give a
final concentration in the milk of 0,2 % and, in such cases, the ratio
between the amount of milk and the solution of mercuric chloride must
be 10 to 1.1997L0012 — EN — 31.12.1998 — 001.001 — 28
▼B
5. The reaction must be carried out using one of the following methods:
— on a column of milk at least 25 mm high and on a volume of milk of
1 ml to which 0,03 ml of one of the standardized stained antigens has
been added,
— on a column of milk at least 25 mm high and on a volume of milk of
1 ml to which 0,05 ml of one of the standardized stained antigens has
been added,
— on a volume of milk of 8 ml to which 0,08 ml of one of the standar-
dized stained antigens has been added,
— on a column of milk at least 25 mm high and on a volume of milk of
2 ml to which 0,05 ml of one of the standardized stained antigens has
been added.
6. The mixture of milk and antigens must be incubated at 37ºC for not less
than 45 minutes and not more than 60 minutes. The test must be assessed
within 15 minutes of removal from the incubator.
7. The reaction must be assessed according to the following criteria:
(a)negative reaction: coloured milk, colourless cream;
(b)positive reaction: milk and cream identically coloured or colourless
milk and coloured cream.
D.Thebufferedbrucellaantigentest
The buffered brucella antigen test may be carried out using one of the
following methods:
(a)Manual test
1. The standard serum shall be the second international standard anti-
brucella abortus serum which is supplied by the Veterinary Labora-
tory Agency, Addlestone, Weybridge, England.
2. The antigen shall be prepared without reference to the cell concentra-
tion, but its sensitivity must be standardized in relation to the second
international standard anti-brucella abortus serum in sucha way th atthe antigen produces a positive reaction with serum dilution of 1:47,5
and a negative reaction witha dilution of 1:55.
3. The antigen shall be suspended in buffered brucella antigen diluent at
a pH of 3,65 ± 0,5 and may have been stained by the use of rose
bengal dye.
4. Weybridge strain No 99 or USDA 1119 or any other strain of equiva-
lent sensitivity must be used for preparing the antigen.
5. The culture media used for keeping the strain in the laboratory and
for producing the antigen must be such that they do not encourage
bacterial dissociation (S-R); potato agar medium or continuous
culture methods should be used.
6. The antigen shall be tested against eight freeze-dried known positive
and negative sera.
7. The official supervision and control of standard serum and antigen
shall be carried out by the official bodies listed in A.9 of this Annex.
8. The antigen shall be delivered ready for use.9. The buffered brucella antigen test shall be carried out in the
following manner:
(a) one drop (0,03 ml) of antigen should be placed alongside one
drop (0,03 ml) of the serum on a white plate;
(b) they should be mixed with an applicator stick, first in a straight
line and then in a circle of about 10 to 12 mm diameter;
(c) the plate should then be rocked back and forth for four minutes
(about 30 times per minute);
(d) readings should be taken in a good light; if there is no evidence
of agglutination, the test shall be regarded as negative; any
degree of agglutination shall be regarded as positive, unless there
has been excessive drying round the edges.
(b)Automated method
The automated method must be at least as sensitive and accurate as the
manual method.1997L0012 — EN — 31.12.1998 — 001.001 — 29
▼B
E.Plasmaringtest
(a)Extraction of the plasma
The tube containing blood, coagulation of which has been inhibited by
the addition of EDTA, should be centrifuged for three minutes at
3 000 r/min and subsequently kept at 37ºC for 12 to 24 hours.
(b)Evaluation
0,2 ml of stabilized plasma should be placed in a tube with 1 ml of
untreated milk. After mixing, one drop (0,05 ml) of ABR antigen should
be added and the whole again mixed. The antigen should be standar-
dized in relation to a standard antigen supplied by the body referred to
in A.9 (a).
Following an incubation period of 45 minutes at 37ºC, a reading should
be taken within 15 minutes. The result shall be regarded as positive if
the colour of the ring has become the same as, or darker than, that of
the milk column.
F.Plasmaagglutination
The plasma extracted in accordance with E (a) may be used immediately
after centrifuging, no thermal stabilization being necessary. 0,05 ml of
plasma should be mixed with 1 ml of antigen for 50 % serum agglutination,
which corresponds to a dilution of 1:20 for serum agglutination. A reading
should be taken after 18 to 24 hours incubation at 37ºC. 50 % or more
agglutination shall be regarded as positive.
G.Micro-agglutinationtest
1. Diluents are made up of 0,85 % physiological saline solution phenolized
at 0,5 %.
2. The antigen shall be prepared as described under A.6, 7 and 8 and shall
be titrated as described under A.5. At the moment the antigen is used,
safranin O shall be added at 0,02 % (final dilution).
3. The standard serum is the same as that under A.1.4. The standard serum must be supplied by the Bundesgesundheitsamt,
Berlin.
5. The micro-agglutination test shall be carried out on plates bearing wells
with conical bottoms of a volume of 0,250 ml. The test shall be carried
out as follows:
(a) predilution of the serum: 0,050 ml of each serum to be tested are
added to eachwell containing 0,075 ml of diluent. Th e mixtures are
shaken for 30 seconds;
(b) gradual serum dilution: prepare at least three dilutions for each
serum. To this end from the predilutions (1:2,5) 0,025 ml of each
serum is taken and transferred to a well containing 0,025 ml of
diluent. In this way the first dilution reaches a strength of 1:5 and
the following dilutions are carried out by doubling;
(c) addition of antigen: 0,025 ml of antigen is added to eachwell
containing the different serum dilutions. After being shaken for 30
seconds the plates are closed with their respective lids and kept at
37ºC for 20 to 24 hours in a humidified atmosphere;
(d) reading the results: assessment of the aspect of the sedimentation of
the antigen is made by examining the bottom of the well reflected in
a concave mirror placed above it. If there is a negative reaction, the
antigen forms a sediment in the form of a compact button with clear
edges and having an intense red colour. If there is a positive reaction
on the other hand, a diffused pink veil is formed that is evenly
distributed. The different percentages of agglutination are determined
by comparison withantigen ch ecks indicating 0, 25, 50, 75 and 100 %
agglutination. The title of each serum is expressed in international
units of agglutination per ml. Controls withnegative and positive
serum diluted so as to contain 30 international units of agglutination
per ml should be included in the test.1997L0012 — EN — 31.12.1998 — 001.001 — 30
▼B
H.Enzyme-linkedimmunosorbentassay(Elisa)fordetectingbovinebrucel-
losis
1. The material and reagents to be used are as follows:
(a) solid-phase microplates, cuvettes or any other solid phase;
(b) the antigen is fixed to the solid phase with or without the aid of poly-
clonal or monoclonal catching antibodies;
(c) the biological fluid to be tested;
(d) a corresponding positive and negative control;(e) conjugate;(f) a substrate adapted to the enzyme used;
(g) a stopping solution, if necessary;
(h) solutions for the dilution of the test samples for preparations of the
reagents and for washing;
(i) a reading system appropriate to the substrate used.
2.Standardization and sensitivityof test
(1) Bulk milk samples are classified negative if they give a reaction less
than 50 % of that given by a 1 in 10000 dilution of the second inter-
national brucellosis standard serum made up in negative milk.
(2) Individual serum samples are classified negative if they give a reac-
tion less than 10 % of that given by a 1 in 200 dilution of the second
international brucellosis standard serum made up in saline solution or
in any other recognized dilution, in accordance with the procedure
laid down in Article 17 after receiving the opinion of the Scientific
Veterinary Committee.
The brucellosis Elisa standards shall be as specified in A.1 and 2 (to
be used at the dilutions indicated on the label).
3.Conditions for use of the Elisa test for bovine brucellosis
The Elisa method may be used on a sample of milk or whey taken from
the milk collected from a farm with at least 30 % of dairy cows in milk.
If this method is used, measures must be taken to ensure that the samples
taken can be identified with the animals from which the milk or sera
examined were taken.1997L0012 — EN — 31.12.1998 — 001.001 — 31
▼B
ANNEX D
CHAPTER I
OFFICIALLY ENZOOTIC-BOVINE-LEUKOSIS-FREE HERDS,
MEMBERSTATESANDREGIONS
A. Officially enzootic-bovine-leukosis-free herd means a herd in which:
(i) there is no evidence, either clinical or as a result of a laboratory test, of
any case of enzootic bovine leukosis in the herd and no such case has
been confirmed in the previous two years; and
(ii) all animals over 24 months of age have reacted negatively during the
preceding 12 months to two tests carried out in accordance with this
Annex, at an interval of at least four months; or
(iii) it meets the requirements of (i) above and is situated in an officially
enzootic-bovine-leukosis-free Member State or region.
B. An individual herd shall retain officially enzootic-bovine-leukosis-free status
provided:
(i) the condition in A (i) continues to be fulfilled;
(ii) any animals introduced into the herd come from an officially enzootic-
bovine-leukosis-free herd;
(iii) all animals over 24 months of age continue to react negatively to a test
carried out in accordance with Chapter II at intervals of three years.
C. The officially leukosis-free status of a herd shall be suspended if the condi-
tions detailed in B are not fulfilled.
D. The status shall remain suspended until the following requirements are
complied with:
1. If a single animal in an officially enzootic-bovine-leukosis-free herd has
reacted positively to one of the tests referred to in Chapter II:
(i) the animal which has reacted positively, and, in the case of a cow,
any calf it may have produced, must have left the herd for slaughter
under the supervision of the veterinary authorities;
(ii) the remaining animals have reacted negatively to a serological test
carried out in accordance with Chapter II three months at least after
removal of the positive animal and any possible progeny thereof;
(iii) an epidemiological inquiry must be conducted and the herds linked
epidemiologically to the infected herd must be subjected to the
measures laid down in (ii).
However, the competent authority may grant a derogation from the obli-
gation to slaughter the calf of an infected cow where it was separated
from its mother after calving. In this case, the calf must be made subject
to the requirements provided for in 2 (iii).
2. Where more than one animal from an officially enzootic-bovine-leukosis-
free herd has reacted positively or where infection has been confirmed in
a herd:
(i) the animals which have reacted positively and, in the case of cows,
their calves, must be removed for slaughter under the supervision of
the veterinary authorities;
(ii) all animals aged over 24 months must react negatively to two tests
carried out in accordance withCh apter II at an interval of at least
four months and no more than 12 months;
(iii) all other animals must, after identification, remain on the holding
until they are aged over 24 months and have satisfied the tests
referred to in (ii);
(iv) an epidemiological inquiry must be conducted, and the herds linked
epidemiologically to the infected herd must be subjected to the
measures laid down in (ii).
However, the competent authority may grant a derogation from the obli-
gation to slaughter the calf of an infected cow where it was separated
from its mother after calving. In this case, the calf must be made subject
to the requirements provided for in 2 (iii).
3. Where the officially enzootic-bovine-leukosis-free status of a herd has
been suspended for any other reason, all animals in the herd aged over
24 months must give a negative reaction to a serological test carried out
in accordance withCh apter II.1997L0012 — EN — 31.12.1998 — 001.001 — 32
▼B
E. In accordance with the procedure in Article 17, the Commission may
propose that a Member State or region of a Member State may become offi-
cially enzootic-bovine-leukosis-free if:
(a) at least 99,8 % of the bovine herds are officially enzootic bovine
leukosis free herds within the meaning of A;
or
(b) no case of enzootic bovine leukosis has been confirmed in the Member
State or region for the past three years and
in the case of a Member State, all animals aged over 24 months in at
least 10 % of herds, selected randomly, have been tested with negative
results in accordance with Chapter II in the previous 24 months or
in the case of a region or a Member State, all animals aged over 24
months have undergone a test provided for in Chapter II with negative
result.
F. A Member State or a region of a Member State shall retain officially
enzootic-bovine-leukosis-free status if:
(i) every year either a random sample with a confidence rating of 99 % has
established that less than 0,2 % of the herds were infected or not less
than 20 % of bovine animals over two years of age have been tested
and have reacted negatively to a test carried out in accordance with
Chapter II;
or
(ii) where no case of enzootic bovine leukosis has been recorded in the
Member State or region in a proportion of one herd out of 10000 for
at least three years, a decision may be taken in accordance with Article
17 to cease routine serological testing provided that:
— all cattle slaughtered within the territory of that Member State or
region are submitted to a post-mortem examination by an official
veterinarian who must issue notification of all tumours with a view
to laboratory examination, and
— the Member State shall report the occurrence of all cases of enzootic
bovine leukosis in the region affected by the decision to the
Commission. The Commission may, in accordance with the proce-
dure laid down in Article 17, propose that the decision to cease
routine serological testing be suspended or revoked, and
— any cattle which react positively to an immune-diffusion test are
slaughtered and the herd remains subject to restrictions until re-
establishment of its status pursuant to Chapter I.D.
G. (i) The officially enzootic-bovine-leukosis-free status of a Member State or
region of a Member State shall be suspended, in accordance with the
procedure in Article 17, if enzootic bovine leukosis is detected and
confirmed in more than 0,2 % of holdings in the region or Member
State.
(ii) The officially enzootic-bovine-leukosis-free status may be restored, in
accordance withth e procedure in Article 17, if:
(a) in addition to the measures provided for in paragraphs D.1 and 2, at
least 20 % of the other herds, selected randomly, in the region or
Member State have, within a 12-month period, undergone one of
the tests referred to in Chapter II;
(b) the results of this testing establish, with a confidence rating of 99 %,
that less than 0,2 % of herds are infected.
CHAPTER II
TESTSFORENZOOTICBOVINELEUKOSIS
Tests for enzootic bovine leukosis shall be carried out by the immune-diffusion
test under the conditions described in A and B or by the enzyme-linked immu-
nosorbent assay (Elisa) under the conditions described in C. The immune-
diffusion method may only be used for individual tests. If test results are the
subject of a duly-substantiated challenge, an additional check shall be carried
out by means of the immune-diffusion test.
A.Agargelimmune-diffusiontestforenzootic bovineleukosis
1. The antigen to be used in the test must contain bovine leukosis virus
glycoproteins. The antigen must be standardized against a standard
serum (El serum) supplied by the State Veterinary Serum Laboratory,
Copenhagen.1997L0012 — EN — 31.12.1998 — 001.001 — 33
▼B
2. The official institutes indicated below must be made responsible for
calibrating the standard working antigen of the laboratory against the
official EEC standard serum (El serum) provided by the State Veterinary
Serum Laboratory, Copenhagen.
(a) Germany: Bundesforschungsanstalt für Viruskrankheiten der Tiere,
Tubingen;
(b) Belgium: Institut national de recherches vétérinaires, Brussels;
(c) France: Laboratoire national de pathologie bovine, Lyon;(d) Grand Duchy of Luxembourg:
(e) Italy: Istituto zooprofilattico spenmentale, Perugia;
(f) Netherlands: Instituut voor Dierhouderij en Diergezondheid (ID-
DLO), Lelystad;
(g) Denmark: Statens Veterinaere Serum Laboratorium, Copenhagen;(h) Ireland: Veterinary Research Laboratory, Abbotstown, Dublin;
(i) United Kingdom:
(1) Great Britain: Veterinary Laboratory Agency, Addlestone,
Weybridge, England;
(2) Northern Ireland: Veterinary Research Laboratory, Stormont,
Belfast;
(j) Spain: Subdirreccion general de sanidad animal. Laboratorio de
sanidad y produccion animal Algete, Madrid;
(k) Portugal: Laboratorio Nacional de Investigaçáo Veterinária, Lisbon;
(l) Greece: Κέντρο Κτηνιατρικών Ιδρυµάτων Νεαπόλεως 25, 153
10 Αθήνα;;
(m) Austria: Bundesanstalt für Tierseuchenbekämpfung, Mödling;
(n) Finland: Eläinlääkintä-ja elintarvikelaitos, Helsinki/Anstalten för
veterinärmedicin ochlivsmedel, Helsingfors;
(o) Sweden: Statens veterinärmedicinska anstalt, Uppsala.
3. The standard antigens used in the laboratory must be submitted at least
once a year to the EEC reference laboratories listed in 2 for testing
against the official EEC standard serum. Apart from this standardization,
the antigen in use can be calibrated in accordance with B.
4. The reagents for the test shall consist of:
(a) antigen: the antigen must contain specific glycoproteins of enzootic
bovine leukosis virus which has been standardized against the offi-
cial EEC serum;
(b) the test serum;
(c) known positive control serum;
(d) agar gel:
0,8 % agar,
8,5 % NaCl,
0,05 M Tris-buffer pH 7,2;
15 ml of this agar must be introduced into a petri dish of 85 mm
diameter, resulting in a depthof 2,6 mm of agar.
5. A test pattern of seven moisture-free wells must be cut in the agar to the
bottom of the plate; the pattern must consist of one central well and six
wells in a circle around it.
Diameter of central well: 4 mmDiameter of peripheral wells: 6 mmDistance between central and peripheral wells: 3 mm
6. The central well must be filled with the standard antigen. Peripheral
wells 1 and 4 (see diagram below) are filled withth e known positive
serum, wells 2, 3, 5 and 6 with the test sera. The wells must be filled
until the meniscus disappears.
7. This results in the following quantities being obtained:
antigen: 32 µl
control serum: 73 µltest serum: 73 µl.1997L0012 — EN — 31.12.1998 — 001.001 — 34
▼B
8. Incubation must be for 72 hours at room temperature (20 to 27ºC) in a
closed humid chamber.
9. The test may be read at 24 and 48 hours but a final result may not be
obtained before 72 hours:
(a) a test serum is positive if it forms a specific precipitin line withth e
BLV antigen and forms a complete line of identity withth e control
serum;
(b) a test serum is negative if it does not form a specific precipitin line
with the BLV antigen and if it does not bend the line of the control
serum;
(c) the reaction cannot be considered conclusive if it:
(i) bends the line of the control serum towards the BLV antigen
well without forming a visible precipitin line with the antigen;
or
(ii) if it cannot be read either as negative or as positive.
In inconclusive reactions the test may be repeated and concentrated
serum utilized.
10. Any other well configuration or pattern may be utilized provided that
the E4 serum diluted 1:10 in negative serum can be detected as positive.
B.Methodforantigenstandardization
Solutions and materials required1. 40 ml of 1,6 % agarose in 0,05 M Tris/HCl buffer, pH 7,2 with8,5 %
NaCl;
2. 15 ml of a bovine leukosis serum, having antibody only to bovine
leukosis virus glycoproteins, diluted 1:10 in 0,05 M Tris/HCl buffer, pH
7,2 with8,5 % NaCl;
3. 15 ml of a bovine leukosis serum, having antibody only to bovine
leukosis virus glycoproteins, diluted 1:5 in 0,05 M Tris/HCl buffer, pH
7,2 with8,5 % NaCl;
4. four plastic petri dishes with a diameter of 85 mm;5. a punchwitha diameter of 4 to 6 mm;6. a reference antigen;7. the antigen which is to be standardized;
8. a water bath(56ºC).
ProcedureDissolve the agarose (1,6 %) in the Tris/HCl buffer by carefully heating to
100ºC. Place in 56ºC water bath for approximately one hour. Also, place the
bovine leukosis serum dilutions in a 56ºC water bath.
Now mix 15 ml of the 56ºC agarose solution with the 15 ml bovine leukosis
serum (1:10), quickly shake and pour 15 ml into each of two petri dishes.
Repeat this procedure with the bovine leukosis serum diluted 1:5.
When the agarose has hardened, holes are made in it as follows:Addition of antigen
(i) Petri dishes 1 and 3:
well A — undiluted reference antigen,
well B — 1:2 diluted reference antigen,wells C and E — reference antigen,well D — undiluted test antigen.
(ii) Petri dishes 2 and 4:
well A — undiluted test antigen,
well B — 1:2 diluted test antigen,well C — 1:4 diluted test antigen,well D — 1:8 diluted test antigen.
Additional instructions1. The experiment shall be carried out with two serum dilutions (1:5 and
1:10) in order to achieve optimal precipitation
2. If the precipitation diameter is too small with both dilutions, then the
serum must be further diluted.1997L0012 — EN — 31.12.1998 — 001.001 — 35
▼B
3. If the precipitation diameter in both dilutions is too large and faint, then a
lower serum must be chosen.
4. The final concentration of the agarose must be 0,8 %; that of the sera 5
and 10 % respectively.
5. Plot the measured diameters in the following coordinate system. The dilu-
tion of the antigen to be tested with the same diameter as the reference
antigen is the working dilution.
C.Enzyme-linked immunosorbent assay (Elisa) for detecting enzootic
bovineleukosis
1. The material and reagents to be used are as follows:
(a) solid-phase microplates, cuvettes or any other solid phase;
(b) the antigen is fixed to the solid phase with or without the aid of poly-
clonal or monoclonal catching antibodies. If antigen is coated directly
to the solid phase, all test samples giving positive reactions have to
be retested against control antigen in the case of EBL. The control
antigen should be identical to the antigen except for the BLV anti-
gens. If catching antibodies are coated to the solid phase, the
antibodies must not react to antigens other than BLV antigens;
(c) the biological fluid to be tested;
(d) a corresponding positive and negative control;
(e) conjugate;(f) a substrate adapted to the enzyme used;(g) a stopping solution, if necessary;
(h) solutions for the dilution of the test samples for preparations of the
reagents and for washing;
(i) a reading system appropriate to the substrate used.
2.Standardization and sensitivityof test
The sensitivity of the Elisa assay must be of such a level that E4 serum is
scored positive when diluted 10 times (serum samples) or 250 times (milk
samples) more than the dilution obtained of individual samples when
these are included in pools. In assays where samples (serum and milk)
are tested individually E4 serum diluted 1 to 10 (in negative serum) or 1
to 250 (in negative milk) must be scored positive when tested in the same
assay dilution as used for the individual test samples. The official insti-tutes listed in A.2 will be responsible for checking the quality of the
Elisa method, and in particular for determining, for each production
batch, the number of samples to be pooled on the basis of the count
obtained for the E4 serum.
The E4 serum will be supplied by the National Veterinary Laboratory,
Copenhagen.
3.Conditions for use of the Elisa test for EBL
The Elisa method may be used on a sample of milk or whey taken from
the milk collected from a farm with at least 30 % of dairy cows in milk.
If this method is used, measures must be taken to ensure that the samples
taken can be identified with the animals from which the milk or sera
examined were taken.1997L0012 — EN — 31.12.1998 — 001.001 — 36
▼B
ANNEX E (I)
(a)Bovinediseases
— Foot-and-mouthdisease
— Rabies— Tuberculosis— Brucellosis— Contagious bovine pleuropneumonia— Enzootic bovine leukosis— Anthrax
(b)Swinediseases
— Rabies
— Brucellosis— Classical swine fever— African swine fever— Foot-and-mouthdisease— Swine vesicular disease— Anthrax1997L0012 — EN — 31.12.1998 — 001.001 — 37
▼B
ANNEX E (II)
— Aujeszky's disease
— Infectious bovine rhinotracheitis— Brucella suis infection— Transmissible gastro-enteritis1997L0012 — EN — 31.12.1998 — 001.001 — 38
▼B
ANNEX F
1997L0012 — EN — 31.12.1998 — 001.001 — 39
▼B
1997L0012 — EN — 31.12.1998 — 001.001 — 40
▼B
1997L0012 — EN — 31.12.1998 — 001.001 — 41