Commission Regulation (EC) No 1248/2001 of 22 June 2001 amending Annexes III, X and XI to Regulation (EC) No 999/2001 of the European Parliament and of the Council as regards epidemio-surveillance and testing of transmissible spongiform encephalopathies
Date: 2001-06-22
Institutions: European Commission
Subjects: animal disease, bovine spongiform encephalopathy, disease prevention, epidemiology, public health, veterinary inspection

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COMMISSION REGULATION (EC) No 1248/2001

of 22 June 2001

amending Annexes III, X and XI to Regulation (EC) No 999/2001 of the European Parliament and of
the Council as regards epidemio-surveillance and testing of transmissible spongiform encephalo-
pathies

THE COMMISSION OF THE EUROPEAN COMMUNITIES,

not excluded). Greece did not submit a dossier for
assessment, referring to legal and technical uncertainties.

Having regard to the Treaty establishing the European
Community,

(5)

Having regard to Regulation (EC) No 999/2001 of the Euro-
pean Parliament and of the Council of 22 May 2001 laying
down rules for the prevention, control and eradication of
certain transmissible spongiform encephalopathies (1), and in
particular Articles 20(2) and 23 thereof,

Whereas:

(1)

(2)

(3)

(4)

Detailed rules for monitoring of transmissible spongi-
form encephalopathies (TSEs)
in bovine, ovine and
caprine animals are laid down in Annex III to Regulation
(EC) No 999/2001. These rules
include systematic
testing of bovine animals over 30 months of age
entering the food chain and random testing of bovine
animals over 30 months of age not entering the food
chain. In addition, all bovine animals subject to casualty
slaughter or found sick when slaughtered for destruction
under the over thirty months scheme (OTMS) shall be
tested. Ovine and caprine animals with clinical signs
compatible with TSE shall be subject to active surveil-
lance.

In view of the detection of bovine spongiform encephal-
opathy (BSE) in two bovine animals at the age of 28
months
in routine testing of casualty slaughtered
animals and to provide an early warning system of the
emergence of any unfavourable trends in BSE incidence
in younger animals, the age limit should be reduced to
24 months in animals belonging to certain risk popula-
tions.

In the monitoring carried out during the first trimester
of 2001, positive BSE cases were detected in all Member
States except in Greece, Luxembourg, Austria, Finland
and Sweden. The number of bovine animals belonging
to certain risk groups tested in the above Member States
were: 248 in Greece, 763 in Luxembourg, 3 295 in
Austria, 4 527 in Finland and 8 254 in Sweden.

In its opinion of 6 July 2000 on the geographical risk of
the Scientific Steering Committee (SSC)
BSE (GBR),
concluded that the GBR level of Luxembourg was III
(BSE confirmed at a low level) and the GBR level of
Austria, Finland and Sweden was II (BSE unlikely, but

(1) OJ L 147, 31.5.2001, p. 1.

In the light of the monitoring carried out in Austria,
Finland and Sweden and of the assessment of the SSC,
the presence of BSE in those Member States is unlikely,
but not excluded. If present, BSE would most likely be
detected by examining bovine animals which have died
on farms, which have been subject to casualty slaughter
or have been found sick at normal slaughter. Those
Member States should therefore be allowed to reduce the
testing in healthy slaughtered bovine animals.

(6) With a view to additional information of the occurrence
of BSE in the United Kingdom, the testing in the OTMS
should be expanded to include all animals born within
one year after the effective enforcement of the feed ban.
Other bovine animals slaughtered under the OTMS
should be tested at random.

(7)

(8)

(9)

(10)

(11)

Member States should be allowed to test other bovine
animals on a voluntary basis, in particular where those
animals are considered to present a higher risk, provided
it is done without disrupting trade.

It is necessary to clarify the measures following testing
of bovine animals and to introduce measures to prevent
carcases potentially
test-positive
carcases from entering the food chain.

contaminated

by

Rapid post-mortem testing should be introduced on a
random basis to improve the detection of scrapie in
ovine and caprine animals. To obtain a more complete
picture of the situation,
it is necessary to carry out
random sampling in two different target populations:
dead-on-farm animals and slaughtered animals.

In Member States with small national herds of sheep and
goats it is difficult to carry out statistically meaningful
sampling in both target groups. Those Member States
should therefore be allowed to use a smaller sample size
but targeted at animals where the likelihood to find
positive cases is highest.

In view of the role of genetic resistance in the develop-
ment of clinical scrapie and the possibility to use
breeding programmes in the prevention, control and
eradication of scrapie, the genotype of all scrapie cases
should be determined and cases found in resistant geno-
types should be submitted for strain-typing.

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(12)

The list of national reference laboratories should be
updated.

HAS ADOPTED THIS REGULATION:

(13)

Following the introduction of rapid tests in monitoring
programmes for ovine and caprine animals it is neces-
sary to lay down appropriate diagnostic methods and
protocols. Furthermore,
the diagnostic methods and
protocols laid down for bovine animals should be
updated.

(14) According to Article 22 of Regulation (EC) No 999/
2001, a conclusive statistical survey shall be used to
confirm or overturn the risk analysis conclusions carried
out as the first step in determining the BSE status of a
country or a region. The minimum criteria for the stat-
istical survey are laid down in Part B of Annex XI. In
view of
the lower BSE risk in Austria, Finland and
Sweden, as assessed by the SSC, and the dispropor-
tionate resources
involved, a derogation should be
provided for those Member States to exclude from the
survey dead-on-farm animals in remote areas with a low
animal density.

(15)

In the interest of clarity Commission Decisions 98/
272/EC (1) on epidemio-surveillance for transmissible
spongiform encephalopathies,
amended by
Decision 2001/8/EC (2), and 2000/764/EC (3) on the
testing of bovine animals for the presence of bovine
spongiform encephalopathy, as amended by Decision
2001/8/EC, should be repealed.

last

as

(16)

The measures provided for in this Regulation are in
accordance with the opinion of the Standing Veterinary
Committee,

Article 1

Regulation (EC) No 999/2001 is amended as follows:

1. The text in Annex III is replaced by the text in Annex I to

this Regulation.

2. The text in Annex X, Chapter A, point 3, is replaced by the

text in Annex II to this Regulation.

3. The text in Annex X, Chapter C, is replaced by the text in

Annex III to this Regulation.

4. The text in Annex XI, Chapter B, is replaced by the text in

Annex IV to this Regulation.

Article 2

1.

Decisions 98/272/EC and 2000/764/EC are repealed.

References to the repealed Decisions shall be construed as
2.
references to Regulation (EC) No 999/2001. In particular, refer-
ences to Annex IVA of Decision 98/272/EC shall be construed
as references to Annex X, Chapter C, point 4, to Regulation
(EC) No 999/2001.

Article 3

This Regulation shall enter into force on the day following its
publication in the Official Journal of the European Communities.

It shall apply from 1 July 2001. However, the provisions of
Annex III, Chapter A, section II, to Regulation (EC) No 999/
2001, as set out in Annex I to this Regulation, shall apply from
1 January 2002.

The provisions of Annex III to Regulation (EC) No 999/2001,
as set out in Annex I to this Regulation, shall be reviewed in
the light of the results obtained during the first six months of
the monitoring.

This Regulation shall be binding in its entirety and directly applicable in all Member States.

Done at Brussels, 22 June 2001.

For the Commission

David BYRNE

Member of the Commission

(1) OJ L 122, 24.4.1998, p. 59.
(2) OJ L 2, 5.1.2001, p. 28.
(3) OJ L 305, 6.12.2000, p. 28.

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ANNEX I

‘ANNEX III

MONITORING SYSTEM

CHAPTER A

I. MONITORING IN BOVINE ANIMALS

1. General

Monitoring in bovine animals shall be carried out in accordance with the laboratory methods laid down in
Annex X, Chapter C, point 3.1(b).

2. Monitoring in animals slaughtered for human consumption

2.1. All bovine animals over 24 months of age:

— subject to “special emergency slaughtering” as defined in Article 2(n) of Council Directive 64/433/EEC (1), or
— slaughtered in accordance with Annex I, Chapter VI, point 28(c), to Directive 64/433/EEC,

shall be tested for BSE.

2.2. All bovine animals over 30 months of age subject to normal slaughter for human consumption shall be tested

for BSE.

2.3. By way of derogation from point 2.2, and with regard to bovine animals born, reared and slaughtered on their
territory, Austria, Finland and Sweden may decide to examine only a random sample. The sample shall comprise
at least 10 000 animals per year.

3. Monitoring in animals not slaughtered for human consumption

Bovine animals over 24 months of age which have died or been killed but which were not
— killed for destruction pursuant to Commission Regulation (EC) No 716/962 (2),
— killed in the framework of an epidemic, such as foot-and-mouth disease,
— slaughtered for human consumption,

shall be tested for BSE at random. The number of samples shall not be less than the sample size indicated in the
table. The sampling must be representative for each region and continuous.

Total population
over 24 months

Sample size (*)

Total population
over 24 months

Sample size (*)

100 000

200 000

300 000

400 000

500 000

600 000

700 000

800 000

900 000

1 000 000

1 500 000

2 000 000

2 500 000

3 000 000

3 500 000

4 000 000

950

1 550

1 890

2 110

2 250

2 360

2 440

2 500

2 550

2 590

3 000

3 500

4 000

4 500

5 000

5 500

4 500 000

5 000 000

5 500 000

6 000 000

6 500 000

7 000 000

7 500 000

8 000 000

8 500 000

9 000 000

9 500 000

10 000 000

10 500 000

11 000 000

11 500 000

12 000 000

6 000

6 500

7 000

7 500

8 000

8 500

9 000

9 500

10 000

10 500

11 000

11 500

12 000

12 500

13 000

13 500

(*) The sample size has been calculated to detect a prevalence of 0,1 % with a 95 % confidence in the subpopulation referred to in
point 3, based on the assumption that the proportion of this subpopulation in the total popuolation of bovine animals over
24 months of age is 1 %. Where the size of the total population of bovine animals over 24 months of age is 1 500 animals or
more, the sample size has been increased by 500 samples per 500 000 animals as a proportionality adjustment, to take
account of the larger likelihood of variation in risk for BSE within the population.

(1) OJ 121, 29.7.1964, p. 2012/64.
(2) OJ L 99, 20.4.1996, p. 14.

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4. Monitoring in animals purchased for destruction pursuant to Regulation (EC) No 716/96

4.1. All animals subject to casualty slaughter or found sick at ante-mortem inspection shall be tested for BSE.

4.2. All animals born between 1 August 1996 and 1 August 1997 shall be tested for BSE.

4.3. A random sample comprising at least 50 000 animals annually of animals not covered by points 4.1 or 4.2 shall

be tested for BSE.

5. Monitoring in other animals

In addition to the testing referred to in points 2 to 4, Member States may on a voluntary basis decide to test
other bovine animals on their territory,
in particular where those animals originate from countries with
indigenous BSE, have consumed potentially contaminated feedingstuffs or were born or derived from BSE
infected dams.

6. Measures following testing

6.1. Where an animal slaughtered for human consumption is tested for BSE, the health marking provided for in
Chapter XI of Annex I to Directive 64/433/EEC shall not be carried out on the carcase of that animal until a
negative result to the rapid test has been obtained.

6.2. Member States may derogate from the provisions of point 6.1 where an official system is in place in the
slaughterhouse ensuring that no parts of examined animals bearing the health mark leave the slaughterhouse
until a negative result to the rapid test has been obtained.

6.3. All parts of the body of an animal tested for BSE including the hide shall be retained under official control until
a negative result to the rapid test has been obtained, unless they are destroyed in accordance with Annex V,
point 3 or 4.

6.4. All parts of the body of an animal found positive to the rapid test including the hide shall be destroyed in
accordance with Annex V, point 3 or 4, apart from material to be retained in conjunction with the records
provided for in Chapter B, Section III.

6.5. Where an animal slaughtered for human consumption is found positive to the rapid test, at least the carcase
immediately preceding the test-positive carcase and two carcases immediately following the test-positive carcase
on the same slaughter line shall be destroyed in accordance with point 6.4, in addition to the test-positive
carcase.

6.6. Member States may derogate from the provisions of point 6.5 where a system is in place in the slaughterhouse

preventing contamination between carcases.

II. MONITORING IN OVINE AND CAPRINE ANIMALS

1. General

Monitoring in ovine and caprine animals shall be carried out in accordance with the laboratory methods laid
down in Annex X, Chapter C, point 3.2(b).

2. Monitoring in animals slaughtered for human consumption

Animals over 18 months of age slaughtered for human consumption shall be tested in accordance with the
sample size indicated in the table. The sampling shall be representative for each region and season. The sample
selection shall be designed with a view to avoid the over-representation of any group as regards the origin,
species, age, breed, production type or any other characteristic. The age of the animals shall be estimated based
on dentition, obvious signs of maturity or other reliable information. Multiple sampling in the same flock shall
be avoided, where possible.

Total number of slaughtered
animals over 18 months
of age

Minimum sample size,
slaughtered animals (*)

Total number of slaughtered
animals over 18 months
of age

Minimum sample size,
slaughtered animals (*)

5 000

10 000

15 000

20 000

25 000

30 000

40 000

50 000

4 750

7 760

9 470

10 540

11 270

11 790

12 490

12 940

60 000

70 000

80 000

90 000

100 000

150 000

200 000

13 260

13 490

13 660

13 800

13 910

14 250

14 430

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Total number of slaughtered
animals over 18 months
of age

Minimum sample size,
slaughtered animals (*)

Total number of slaughtered
animals over 18 months
of age

Minimum sample size,
slaughtered animals (*)

250 000

14 540

300 000

14 610

350 000

14 660

400 000

14 700

450 000

14 730

500 000

14 760

600 000

14 790

700 000

14 820

800 000

14 840

900 000

14 850

1 100 000

1 200 000

1 300 000

1 400 000

1 500 000

1 600 000

1 700 000

1 800 000

1 900 000

2 000 000

2 100 000

14 880

14 890

14 890

14 900

14 900

14 910

14 910

14 920

14 920

14 920

14 920

1 000 000

14 870

2 200 000 or more

14 930

(*) The sample size has been calculated to detect a prevalence of 0,02 % with a 95 % confidence in slaughtered animals.

3. Monitoring in animals not slaughtered for human consumption

Animals over 18 months of age which have died or been killed, but which were not:
— killed in the framework of an epidemic, such as foot-and-mouth disease,
— slaughtered for human consumption,

shall be tested in accordance with the sample size indicated in the table. The sampling shall be representative for
each region and season. The sample selection shall be designed with a view to avoid the over-representation of
any group as regards the origin, species, age, breed, production type or any other characteristic. The age of the
animal shall be estimated based on dentition, obvious signs of maturity or other reliable information. Multiple
sampling in the same flock shall be avoided, where possible.

Total number of animals over 18 months of age (*)

Minimum sample size, dead animals (**)

100 000

200 000

300 000

400 000

500 000

600 000

700 000

800 000

900 000

1 000 000

1 500 000 or more

950

1 550

1 890

2 110

2 250

2 360

2 440

2 500

2 550

2 590

3 000

(*) Where the total number of ovine and caprine animals over 18 months of age is not known, the total number of “ewes and

ewe-lambs put to the ram” and “goats which have already kidded and goats mated” shall be used instead.

(**) The sample size has been calculated to detect a prevalence of 0,1 % with a 95 % confidence in dead animals, based on the
assumption that the proportion of dead animals in the total population of ovine and caprine animals over 18 months of age
is 1 %.

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4. Monitoring in Member States with a small ovine and caprine population

Member States, where the total number of ovine and caprine animals over 18 months of age is 500 000 or less
may, by way of derogation from the sampling foreseen in points 2 and 3, decide to monitor the following
combined subpopulation:

(a) animals over 18 months of age which have died or been killed, but which were not:

— killed in the framework of an epidemic, such as foot-and-mouth disease,

— slaughtered for human consumption (“dead animals”); and

(b) animals over 18 months of age whose appearance suggests a chronic wasting condition (“chronic wasting

animals”).

The number of samples tested annually in each Member State from the above combined subpopulation shall not
be less than the sample size indicated in the table.

The sampling shall be representative for each region and season. The sample selection shall be designed with a
view to avoid the over-representation of any group as regards the origin, species, age, breed, production type or
any other characteristic. Where dead animals are sampled, the age of the animal shall be estimated based on
dentition, obvious signs of maturity or other reliable information. Where chronic wasting animals are sampled,
only animals inspected by an official veterinarian and for which the age and clinical symptoms are well
documented shall be selected for sampling. Multiple sampling in the same flock shall be avoided, where possible.

Total number of ovine and caprine animals
over 18 months of age (*)

Minimum sample size, dead and chronic wasting animals

10 000

20 000

30 000

40 000

50 000

60 000

70 000

80 000

90 000

100 000

200 000

300 000

400 000

500 000

100

200

300

400

500

600

700

800

900

950

1 550

1 890

2 110

2 250

(*) Where the total number of ovine and caprine animals over 18 months of age is not known, the total number of “ewes and

ewe-lambs put to the ram” and “goats which have already kidded and goats mated” shall be used instead.

5. Monitoring in other animals

In addition to the monitoring programmes set out in points 2 to 4, Member States may on a voluntary basis
carry out monitoring in other animals,

in particular:

— animals used for dairy production,

— animals originating from countries with indigenous TSEs,

— animals which have consumed potentially contaminated feedingstuffs,

— animals born or derived from TSE infected dams,

— animals from flocks infected with TSE.

6. Measures following testing of ovine and caprine animals

All parts of the body of a tested animal including the hide shall be retained under official control until a negative
result to the rapid test has been obtained, unless they are destroyed in accordance with Annex V, point 3 or 4.

All parts of the body of an animal found positive to the rapid test including the hide shall be destroyed in
accordance with Annex V, point 3 or 4, apart from material to be retained in conjunction with the records
provided for in Chapter B, section III.

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7. Genotyping

The prion protein genotype shall be determined for each positive TSE case in sheep. TSE cases found in resistant
genotypes (sheep of genotypes which encode alanin on both alleles at codon 136, arginin on both alleles at
codon 154 and arginin on both alleles at codon 171) shall immediately be reported to the Commission. Where
possible, such cases shall be submitted for strain-typing. Where strain-typing of such cases is not possible, the
herd of origin and all other herds where the animal has been shall be subjected to enhanced monitoring with a
view to find other TSE cases for strain-typing.

I.

INFORMATION TO BE PRESENTED BY MEMBER STATES IN THEIR REPORT

CHAPTER B

1.

2.

3.

4.

5.

6.

7.

The number of suspected cases per animal species placed under movement restrictions in accordance with
Article 12(1).

The number of suspected cases per animal species subject to laboratory examination in accordance with Article
12(2) and the outcome of the examination.

The number of flocks where suspected cases in ovine and caprine animals have been reported and investigated
pursuant to Article 12(1) and (2).

The estimated size of each subpopulation referred to in Chapter A, Section I, points 3 and 4.

The number of bovine animals tested within each subpopulation referred to in Chapter A, Section I, point 2 to
5, the method for sample selection and the outcome of the tests.

The estimated size of those subpopulations referred to in Chapter A, Section II, points 2 to 4, which have been
selected for sampling.

The number of ovine and caprine animals and flocks tested within each subpopulation referred to in Chapter A,
Section II, points 2 to 5, the method for sample selection and the outcome of the tests.

8. Number, age distribution and geographical distribution of positive cases of BSE and scrapie. The country of
origin, if not the same as the reporting country, of positive cases of BSE and scrapie. Number and geographical
distribution of scrapie positive flocks. The year and, where possible, month of birth should be given for each BSE
case.

9.

Positive TSE cases confirmed in animals other than bovine, ovine and caprine animals.

II.

INFORMATION TO BE PRESENTED BY THE COMMISSION IN ITS SUMMARY

The summary shall be presented in a tabled format covering at least the information referred to in Part I for each
Member State.

III. RECORDS

1.

The competent authority shall keep, for seven years, records of:
— the number and types of animals placed under movement restrictions as referred to in Article 12(1),
— the number and outcome of clinical and epidemiological
investigations as referred to in Article 12(1),
— the number and outcome of laboratory examinations as referred to in Article 12(2),
— the number, identity and origin of animals sampled in the framework of the monitoring programmes as

referred to in Chapter A and, where possible, age, breed and anamnestic information,

— the prion protein genotype of positive TSE cases in sheep,
— where chronic wasting ovine and caprine animals have been selected for sampling, the method to establish

the age of and the clinical symptoms observed in each sampled animal.

2.

The investigating laboratory shall keep, for seven years, all records of testing, in particular laboratory workbooks
and, where appropriate, paraffin blocks and photographs of Western blots.’

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‘3. The national reference laboratories are:

ANNEX II

Austria:

Belgium:

Denmark:

Finland:

France:

Germany:

Greece:

Ireland:

Italy:

Luxembourg:

Netherlands:

Bundesanstalt für Tierseuchenbekämpfung, Mödling
Robert Koch Gasse 17
A-2340 Mödling

CERVA-CODA-VAR
Centre d'Étude et de Recherches Vétérinaires et Agrochimiques
Centrum voor Onderzoek in Diergeneeskunde en Agrochemie
Veterinary and Agrochemical Research Centre
Groeselenberg 99
B-1180 Bruxelles

Danish Veterinary Laboratory
Bülowsvej 27
DK-1790 Copenhagen V

Eläinlääkintä- ja elintarvikelaitos
Hämeentie 57
FIN-00550 Helsinki

Agence Française de Sécurité Sanitaire des Aliments
Laboratoire de pathologie bovine
31, avenue Tony Garnier
BP 7033
F-69342 Lyon Cedex

Bundesforschungsanstalt für Viruskrankheiten der Tiere
Anstaltsteil Insel Riems
Boddenblick 5A
D-17498 Insel Riems

Laboratory of Microbiology and Infectious Diseases
Faculty of Veterinary Medicine
Aristotelian University of Thessaloniki
University Campus
GR-54006 Thessaloniki
(rapid and immunological tests)

Laboratory of Gross Pathology (Morgue
Faculty of Veterinary Medicine
Aristotelian University of Thessaloniki
Giannitson & Voutyra St
GR-54627 Thessaloniki
(histopathology)

Central Veterinary Research Laboratory
Abbotstown
Castleknock
Dublin 15
Ireland

Istituto Zooprofilattico Sperimentale del Piemonte
Liguria e Valle d'Aosta
CEA
Via Bologna
I-148-10150 Torino

CERVA-CODA-VAR
Centre d'Étude et de Recherches Vétérinaires et Agrochimiques
Centrum voor Onderzoek in Diergeneeskunde en Agrochemie
Veterinary and Agrochemical Research Centre
Groeselenberg 99
B-1180 Bruxelles

Instituut voor Dierhouderij en Diergezondheid, ID-DLO Lelystad
Edelhertweg 15
Postbus 65
8200 AB Lelystad
Netherlands

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Portugal:

Spain:

Laboratório Nacional de Investigação Veterinária
Estrada de Benfica, 701
P-1500 Lisboa

Laboratorio de la Facultad de Veterinaria
Departamento de Patología Animal (Anatomía Patológica)
Zaragoza
Spain
(BSE and scrapie, methods other than rapid tests)

Laboratorio Central de Veterinaria de Algete
Madrid
Spain
(rapid tests)

Centro de Investigacion en Sanidad Anímal (CISA)
Crta, De Algete al Casar de Talamanca
28130 Valdeolmos (Madrid)
Spain
(TSEs other than BSE or scrapie)

Sweden:

United Kingdom

National Veterinary Institute
S-751 89 Uppsala

Veterinary Laboratories Agency
Woodham Lane
New Haw
Addlestone
Surrey KT15 3NB
United Kingdom’

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ANNEX III

‘CHAPTER C

Sampling and laboratory testing

1.

Sampling

Any samples intended to be examined for the presence of a TSE shall be collected using the methods and protocols
laid down in the latest edition of the Manual of standards for diagnostic tests and vaccines of the International Office for
Epizooties (IOE/OIE) (hereinafter referred to as “the Manual”). In the absence of such methods and protocols, the
samples shall be collected in a manner appropriate for the correct application of tests. The samples shall be correctly
marked as to the identity of the sampled animal.

2.

Laboratories

Any laboratory examination for TSE shall be carried out in laboratories approved for that purpose.

3. Methods and protocols

3.1. Laboratory testing for the presence of BSE in bovine animals

(a) S u s p e c t c a s e s

Tissues from bovine animals sent for laboratory testing pursuant to the provisions of Article 12(2) shall be
subject to a histopathological examination as laid down in the latest edition of the Manual, except where the
material is autolysed. Where the result of the histopathological examination is inconclusive or negative or where
the material is autolysed, the tissues shall be subjected to an examination by one of the other diagnostic methods
laid down in the Manual (immunocytochemistry, immuno-blotting or demonstration of characteristic fibrils by
electron microscopy). However, rapid tests cannot be used for this purpose.

If the result of one of the above examinations is positive, the animals shall be regarded a positive BSE case.

(b) B S E m o n i t o r i n g

Tissues from bovine animals sent for laboratory testing pursuant to the provisions of Annex III, Chapter A,
Section I (Monitoring in bovine animals) shall be examined by a rapid test.

immediately be subject to
When the result of the rapid test is inconclusive or positive, the tissues shall
confirmatory examinations in an official laboratory. The confirmatory examination shall start by a histopatholog-
ical examination of the brainstem as laid down in the latest edition of the Manual, except where the material is
autolysed or otherwise not suitable for examination by histopathology. Where the result of the histopathological
examination is inconclusive or negative or where the material is autolysed, the tissues shall be subjected to an
examination by one of the other diagnostic methods mentioned under (a).

An animal shall be regarded a positive BSE case if the result of the rapid test is positive or inconclusive, and
— the result of the subsequent histopathological examination is positive, or
— the result of another diagnostic method mentioned under (a) is positive.

3.2. Laboratory testing for the presence of scrapie in ovine and caprine animals

(a) S u s p e c t c a s e s

Tissues from ovine and caprine animals sent for laboratory testing pursuant to the provisions of Article 12(2)
shall be subject to a histopathological examination as laid down in the latest edition of the Manual, except where
the material is autolysed. Where the result of the histopathological examination is inconclusive or negative or
where the material is autolysed, the tissues shall be subjected to an examination by immunocytochemistry or
immuno-blotting, as laid down in the Manual. However, rapid tests cannot be used for this purpose.

If the result of one of the above examinations is positive, the animal shall be regarded a positive scrapie case.

(b) S c r a p i e m o n i t o r i n g

Tissues from ovine and caprine animals sent for laboratory testing pursuant to the provisions of Annex III,
Chapter A, Section II (Monitoring in ovine and caprine animals) shall be examined by a rapid test.

When the result of the rapid test is inconclusive or positive, the brainstem shall immediately be sent to an official
laboratory for confirmatory examinations by immunocytochemistry or immuno-blotting, as referred to under (a).

An animal shall be regarded a positive scrapie case if the result of the confirmatory examination is positive.

L 173/22

EN

Official Journal of the European Communities

27.6.2001

3.3. Laboratory testing for the presence of TSEs other than those referred to in points 3.1 and 3.2

The tests carried out to confirm the suspected presence of a TSE different from those referred to in points 3.1 and
3.2 shall include at least a histopathological examination of brain tissue. The competent authority may also require
laboratory tests such as immunocytochemistry, immuno-blotting, demonstration of characteristic fibrils by electron
microscopy or other methods designed to detect the disease associated form of the prion protein. In any case at least
one other laboratory examination shall be carried out if the initial histopathological examination is negative or
inconclusive. At least three different examinations shall be carried out in the event of the first appearance of the
disease.

In particular, where BSE is suspected in a species other than bovine animals, samples shall be submitted for
strain-typing, where possible.

4. Rapid tests

For the purposes of carrying out the tests in accordance with Article 5(3) and Article 6(1), the following methods
shall be used as rapid tests within the meaning of this Regulation:
— immuno-blotting test based on a western blotting procedure for the detection of the protease-resistant fragment

PrPRes (Prionics Check test),

— chemiluminescent ELISA test involving an extraction procedure and an ELISA technique, using an enhanced

chemiluminescent reagent (Enfer test),

— sandwich immunoassay for PrPRes carried out following denaturation and concentration steps (Bio-Rad Platelia

test).

5. Alternative tests

(To be defined)’

ANNEX IV

‘B. Concerning statistical surveys

1. The statistical survey referred to in Article 22 must cover:

— the animals sampled pursuant to the provisions of Annex III, Chapter A, Section I, points 2.1 and 4.1,
— all the animals in the subpopulation referred to in Annex III, Chapter A, Section I, point 3, instead of a random

sample.

This provision, applicable for a period of one year, may be reviewed in the light of experience gained in the first
six months.

2. Austria, Finland and Sweden may decide to derogate from the provisions of point 1, second indent, in remote

areas with a low animal density.’