Commission Regulation (EC) No 162/2009 of 26 February 2009 amending Annexes III and X to Regulation (EC) No 999/2001 of the European Parliament and of the Council laying down rules for the prevention, control and eradication of certain transmissible spongiform encephalopathies (Text with EEA relevance)
Date: 2009-02-26
Institutions: European Commission
Subjects: animal disease, animal nutrition, animal protein, cattle, derogation from EU law, disease prevention, veterinary legislation

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COMMISSION REGULATION (EC) No 162/2009

of 26 February 2009

amending Annexes III and X to Regulation (EC) No 999/2001 of the European Parliament and of the
laying down rules for the prevention, control and eradication of certain transmissible
Council

spongiform encephalopathies

(Text with EEA relevance)

THE COMMISSION OF THE EUROPEAN COMMUNITIES,

Having regard to the Treaty
Community,

establishing the European

Having regard to Regulation (EC) No 999/2001 of the European
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
the third subparagraph of Article 5(3) and the first paragraph of
Article 23 thereof,

Whereas:

Regulation (EC) No 999/2001 lays down rules for the
prevention, control and eradication of certain trans­
missible spongiform encephalopathies (TSEs) in animals.
It provides that each Member State is to carry out an
annual monitoring programme for TSEs based on active
and passive surveillance.

concerning

Regulation (EC) No 1774/2002 of
the European
Parliament and of
the Council of 3 October 2002,
animal by-
laying down health rules
products not intended for human consumption (2) lays
down the animal and public health rules
the
collection, transport, storage, handling, processing and
use or disposal of animal by-products, to prevent these
products from presenting a risk to animal or public
health.

for

(6)

(7)

(8)

(9)

Article 4(2) of Regulation (EC) No 1774/2002 sets out
the means of disposing of Category 1 material, as defined
in Article 2(1)(b) of that Regulation.

(10)

Part I of Chapter A of Annex III to Regulation (EC) No
999/2001 lays down rules for the monitoring in bovine
animals as well as measures to be applied following
testing of the animals.

According to those rules, all parts of the body of an
tested for bovine spongiform encephalopathy
animal
(BSE),
including the hide, are to be retained under
official control until a negative result to the rapid test
in
has been obtained, unless
in
accordance with two of

they are disposed of
set out
the means

(1)

(2)

(3)

(4)

(5)

Article 4(2) of Regulation (EC) No 1774/2002. Also,
all parts of the body of an animal found positive or
inconclusive to a rapid test including the hide are to
be disposed of in accordance with the same means of
disposal.

Regulation (EC) No 1774/2002 provides for the possi­
bility of additional means of disposal to be approved for
in the light of developments in
Category 1 material,
scientific
are
Such
knowledge.
approved and laid down in Commission Regulation
(EC) No 92/2005 (3).

alternative means

In the interest of consistency of Community legislation,
points 6.3 and 6.4 of Part I of Chapter A of Annex III to
Regulation (EC) No 999/2001 should be amended to
cover also those additional means of disposal.

Chapter C of Annex X to Regulation (EC) No 999/2001
lays down rules for the sampling and laboratory testing
for the presence of TSEs.

According to those rules, the first diagnostic method to
be used for the confirmation of a clinical suspect case of
BSE is based on histopathological examination, which is
the method recommended in an earlier edition of the
Manual for diagnostic tests and vaccines for terrestrial
animals of the World Organisation for Animal Health
(OIE) (‘the Manual’).

In the latest edition of the Manual, adopted in May 2008,
histopathological examination is no longer considered as
the reference diagnostic method for
investigation of
animals suspected of being infected by BSE. According
immunohistochemical and immuno­
to the Manual,
chemical methods,
including rapid tests, may now be
used for
that purpose. The Community reference
laboratory for TSEs considers that applying the same
approach for the investigation of ovine and caprine
animals
suspected of being infected by a TSE is
relevant and scientifically robust.

(1) OJ L 147, 31.5.2001, p. 1.
(2) OJ L 273, 10.10.2002, p. 1.

(3) OJ L 19, 21.1.2005, p. 27.

(11)

The methods and protocols to be used for the BSE active
surveillance in bovine animals
should therefore be
amended to reflect the recent modification of the Manual.

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(12)

(13)

Point 3.2(c) of Chapter C of Annex X to Regulation (EC)
No 999/2001 provides
further examination of
for
positive scrapie cases in ovine and caprine animals in
order to investigate the possible presence of BSE.

cases

(TSE)

In its opinion on classification of atypical Transmissible
Spongiform Encephalopathy
in small
ruminants (1), of 26 October 2005, the European Food
Safety Authority states that atypical scrapie cases are
in its
clearly distinguishable from BSE.
guidelines (2),
the Community reference laboratory for
TSEs considers that if a TSE is confirmed as an atypical
scrapie case, no further testing is required.

In addition,

(14) Diagnosed atypical scrapie cases should therefore be
exempted from the requirement of further examination
set out in point 3.2(c) of Chapter C of Annex X to
Regulation (EC) No 999/2001.

(15)

(16)

(17)

Point 4 of Chapter C of Annex X to Regulation (EC) No
999/2001 sets out a list of rapid tests approved for the
monitoring of TSEs in bovine, ovine and caprine animals.

commercial

The
currently
denomination of
approved TSE tests has recently changed. For the sake
of
these changes should be reflected in
Point 4 of Chapter C of Annex X.

transparency,

some

In addition, the companies manufacturing certain rapid
tests do not exist anymore. Other companies producing
rapid tests have not submitted the details of their quality
system to the Community reference laboratory for
review. Certain other rapid tests have been withdrawn
from the market.

(18)

(19)

It is therefore appropriate to amend accordingly the lists
of rapid tests approved for the monitoring of BSE and
TSEs, set out in point 4 of Chapter C of Annex X to
Regulation (EC) No 999/2001.

In the interest of clarity and legal certainty, the wording
of the heading of point 3.2(c) of Annex X, Chapter C,
should be amended to be consistent with the general
scope of paragraph 3.2 of Annex X, Chapter C which
is related to laboratory testing for the presence of TSE in
ovine and caprine animals.

(20)

Regulation (EC) No 999/2001 should therefore be
amended accordingly.

(21)

The measures provided for in this Regulation are in
Standing
accordance with
Committee on the Food Chain and Animal Health,

opinion

the

the

of

HAS ADOPTED THIS REGULATION:

Article 1

Annexes III and X to Regulation (EC) No 999/2001 are
amended in accordance with the Annex to this Regulation.

Article 2

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

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

Done at Brussels, 26 February 2009.

For the Commission

Androulla VASSILIOU
Member of the Commission

(1) The EFSA Journal (2005) 276, pp. 1-30.
(2) http://www.defra.gov.uk/vla/science/docs/sci_tse_rl_handbookv2

mar07.pdf

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ANNEX

Annexes III and X to Regulation (EC) No 999/2001 are amended as follows:

1.

in Part I of Chapter A of Annex III, points 6.3 and 6.4 are replaced by the following:

‘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
in accordance with
Article 4(2)(a), (b) or (e) of Regulation (EC) No 1774/2002 of the European Parliament and of the Council.

to the rapid test has been obtained, unless they are disposed of

6.4. All parts of the body of an animal found positive or inconclusive to the rapid test including the hide shall be
disposed of in accordance with Article 4(2)(a), (b) or (e) of Regulation (EC) No 1774/2002, apart from material
to be retained in conjunction with the records provided for in Chapter B(III).’;

2.

in Annex X, Chapter C is amended as follows:

(a)

In Point 3.1, points (a) and (b) are replaced by the following:

‘(a) Suspect cases

Samples from bovine animals sent for laboratory testing pursuant to the provisions of Article 12(2) shall
immediately be subjected to confirmatory examinations using at least one of the following methods and
protocols laid down in the latest edition of the Manual:

(i) the immunohistochemical (IHC) method;

(ii) SAF-immunoblot or OIE approved alternative;

(iii) the demonstration of characteristic fibrils by electron microscopy;

(iv) the histopathological examination;

(v) the combination of rapid tests as laid down in the third subparagraph.

In case the histopathological examination is inconclusive or negative, the tissues shall be submitted to a further
examination by one of the other confirmatory methods and protocols.

Rapid tests may be used for both primary screening of suspect cases and, if inconclusive or positive, for
subsequent confirmation, according to the guidelines from the Community reference laboratory and provided
that:

(i) the confirmation is carried out in a national reference laboratory for TSEs; and

(ii) one of the two rapid tests is a Western blot; and

(iii) the second rapid test used:

— includes a negative tissue control and a bovine BSE sample as positive tissue control,

— is of a different type than the test used for the primary screening; and

(iv) if a rapid Western blot is used as the first test, the result of that test must be documented and submitted

to the national reference laboratory for TSEs; and

(v) where the result of the primary screening is not confirmed by the subsequent rapid test, the sample must
be subjected to an examination by one of the other confirmatory methods; in case the histopathological
examination is used for that purpose but proves to be inconclusive or negative, the tissues must be
submitted to a further examination by one of the other confirmatory methods and protocols.

If the result of one of the confirmatory examinations referred to in points (i) to (v) of the first subparagraph is
positive, the animals shall be regarded a positive BSE cases.

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(b) BSE monitoring

Samples from bovine animals sent for laboratory testing pursuant to the provisions of Annex III, Chapter A,
Part I shall be examined by a rapid test.

When the result of the rapid test is inconclusive or positive, the sample shall immediately be subjected to
confirmatory examinations using at least one of the following methods and protocols laid down in the latest
edition of the Manual:

(i) the immunohistochemical (IHC) method;

(ii) SAF-immunoblot or OIE approved alternative;

(iii) the demonstration of characteristic fibrils by electron microscopy;

(iv) the histopathological examination;

(v) the combination of rapid tests as laid down in the fourth subparagraph.

In case the histopathological examination is inconclusive or negative, the tissues shall be submitted to a further
examination by one of the other confirmatory methods and protocols.

Rapid tests may be used for both primary screening and, if inconclusive or positive, for subsequent confir­
mation, according to the guidelines from the Community reference laboratory and provided that:

(i) the confirmation is carried out in a national reference laboratory for TSEs; and

(ii) one of the two rapid tests is a Western blot; and

(iii) the second rapid test used:

— includes a negative tissue control and a bovine BSE sample as positive tissue control,

— is of a different type than the test used for the primary screening; and

(iv) if a rapid Western blot is used as the first test, the result of that test must be documented and submitted

to the national reference laboratory for TSEs; and

(v) where the result of the primary screening is not confirmed by the subsequent rapid test, the sample must
be subjected to an examination by one of the other confirmatory methods; in case the histopathological
examination is used for that purpose but proves to be inconclusive or negative, the tissues must be
submitted to a further examination by one of the other confirmatory methods and protocols.

An animal shall be regarded a positive BSE case if the result of the rapid test is inconclusive or positive, and at
least one of the confirmatory examinations referred to in points (i) to (v) of the first subparagraph is positive.’;

(b) in Point 3.2, point (a) is replaced by the following:

‘(a) Suspect cases

Samples from ovine and caprine animals sent for laboratory testing pursuant to the provisions of Article 12(2)
shall immediately be subjected to confirmatory examinations using at least one of the following methods and
protocols laid down in the latest edition of the Manual:

(i) the immunohistochemical (IHC) method;

(ii) SAF-immunoblot or OIE approved alternative;

(iii) the demonstration of characteristic fibrils by electron microscopy;

(iv) the histopathological examination.

In case the histopathological examination is inconclusive or negative, the tissues shall be submitted to a further
examination by one of the other confirmatory methods and protocols.

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Rapid tests may be used for primary screening of suspect cases. Such tests may not be used for subsequent
confirmation.

Where the result of the rapid test used for primary screening of suspect cases is positive or inconclusive, the
sample shall be subjected to an examination by one of the confirmatory examinations referred to in points (i)
to (iv) of the first subparagraph. In case the histopathological examination is used for that purpose but proves
to be inconclusive or negative, the tissues shall be submitted to a further examination by one of the other
confirmatory methods and protocols.

If the result of one of the confirmatory examinations referred to in point (i) to (iv) of the first subparagraph is
positive, the animals shall be regarded positive TSE cases and further examination as referred to in point (c)
shall be performed.’;

(c)

in point 3.2, the heading of point (c) is replaced by the following:

‘(c) Further examination of positive TSE cases’;

(d) in point 3.2, point (c)(i) is replaced by the following:

‘(i) Primary molecular testing with a discriminatory immuno-blotting

Samples from clinical suspect cases and from animals tested in accordance with Annex III, Chapter A, Part II,
points 2 and 3 which are regarded as positive TSE cases but which are not atypical scrapie cases, following the
examinations referred to in points (a) or (b), or which display characteristics which are deemed by the testing
laboratory to merit investigation, shall be forwarded for further examination by a primary molecular typing
method to:

— Agence Française de Sécurité Sanitaire des Aliments, Laboratoire de pathologie bovine, 31 avenue Tony

Garnier, BP 7033, F-69342, Lyon Cedex, France,

— Veterinary Laboratories Agency, Woodham Lane, New Haw, Addlestone, Surrey KT15 3NB, United

Kingdom, or

— to a laboratory, appointed by the competent authority, which has participated successfully in proficiency
testing organised by the Community reference laboratory for the use of a molecular typing method.’;

(e)

in point 3.2(c)(ii), the word ‘scrapie’ is replaced by ‘TSE’;

(f) point 4 is replaced by the following:

‘4. Rapid tests

For the purposes of carrying out the rapid tests in accordance with Articles 5(3) and 6(1), the following
methods shall be used as rapid tests for the monitoring of BSE in bovine animals:

— the immuno-blotting test based on a Western blotting procedure for the detection of the Proteinase K-

resistant fragment PrPRes (Prionics-Check Western test),

— the chemiluminescent ELISA test involving an extraction procedure and an ELISA technique, using an
test & Enfer TSE Kit version 2.0, automated sample

reagent

(Enfer

enhanced chemiluminescent
preparation),

— the microplate-based immunoassay for the detection of PrPSc (Enfer TSE Version 3),

— the sandwich immunoassay for PrPRes detection with the TeSeE SAP Detection kit carried out following

denaturation and concentration steps with the TeSeE Purification kit (Bio-Rad TeSeE rapid test),

— the microplate-based immunoassay (ELISA) which detects Proteinase K-resistant PrPRes with monoclonal

antibodies (Prionics-Check LIA test),

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— the immunoassay using a chemical polymer for selective PrPSc capture and a monoclonal detection
antibody directed against conserved regions of the PrP molecule (IDEXX HerdChek BSE Antigen Test
Kit, EIA),

— the lateral-flow immunoassay using two different monoclonal antibodies to detect Proteinase K-resistant

PrP fractions (Prionics Check PrioSTRIP),

— the two-sided immunoassay using two different monoclonal antibodies directed against two epitopes

presented in a highly unfolded state of bovine PrPSc (Roboscreen Beta Prion BSE EIA Test Kit),

— the sandwich ELISA for the detection of Proteinase K-resistant PrPSc (Roche Applied Science PrionScreen).

For the purposes of carrying out the rapid tests in accordance with Articles 5(3) and 6(1), the following
methods shall be used as rapid tests for the monitoring of TSE in ovine and caprine animals:

— the sandwich immunoassay for PrPRes detection with the TeSeE SAP Detection kit carried out following

denaturation and concentration steps with the TeSeE Purification kit (Bio-Rad TeSeE rapid test),

— the sandwich immunoassay for PrPRes detection with the TeSeE Sheep/Goat Detection kit carried out
following denaturation and concentration steps with the TeSeE Sheep/Goat Purification kit (Bio-Rad TeSeE
Sheep/Goat rapid test),

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

enhanced chemiluminescent reagent (Enfer TSE Kit version 2.0),

— the microplate-based immunoassay for the detection of PrPSc (Enfer TSE Version 3),

— the immunoassay using a chemical polymer for selective PrPSc capture and a monoclonal detection
antibody directed against conserved regions of the PrP molecule (IDEXX HerdChek BSE-Scrapie Antigen
Test Kit, EIA),

— the immuno-blotting test based on a Western blotting procedure for the detection of the Proteinase K-

resistant fragment PrPRes (Prionics-Check Western Small Ruminant test),

— the microplate-based chemiluminescent immunoassay for the detection of Proteinase K-resistant PrPSc

(Prionics Check LIA Small Ruminants).

In all tests, sample tissue on which the test must be applied must comply with the manufacturer’s instructions
for use.

Producers of rapid tests must have a quality assurance system in place that has been approved by the
Community reference laboratory and ensures that the test performance does not change. Producers must
provide the Community reference laboratory with the test protocols.

Changes to rapid tests and to test protocols may only be made after prior notification to the Community
reference laboratory and provided that the Community reference laboratory finds that the change does not
alter the sensitivity, specificity or reliability of the rapid test. That finding shall be communicated to the
Commission and to the national reference laboratories.’