COMMISSION REGUL ATION (EU) No 1148/2014 
of 28 Oct ober 2014 
amending Annexes II, VII, VIII, IX and X to Regulation (EC) No 999/2001 of the European 
Parliament and of the Council laying down rules for the prev ention, control and eradication of 
certain transmissible spongif orm encephalopathies 
(Text with EEA relevance) 
THE EUR OPEAN COMMISSION, 
Having regar d to the Treaty on the Functioning of the European Union, 
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 spongif orm encephalopathies (1), and in 
particular the first paragraph of Article 23 thereof, 
Whereas: 
(1) Regulation (EC) No 999/2001 lays down rules for the prevention, control and eradication of transmissible spon ­
giform encephalopathies (TSEs) in bovine, ovine and capr ine animals. It applies to the production and placing on 
the mark et of live animals and products of animal origin and in certain specifi c cases to expor ts thereof. 
(2)  Annex II to Regulation (EC) No 999/2001 lays down rules governing the determination of the bovine spongif orm 
encephalopath y (BSE) status of Member States or third countr ies or regions thereof. These rules are based on the 
international standard established by the World Organisation for Animal Health (OIE) in the Terrestr ial Animal 
Health Code (the Code). In the BSE chap ter of the 2013 version of the Code, the expression ‘release assessment ’ 
has been replaced by ‘entr y assessment ’, and the table providing the points targets for a countr y or region has 
been significantly amended to better meet the needs of countr ies with a small or very small cattle population. 
These amendments should be reflected in Annex II. 
(3)  Point 2.2.1 of Chapt er B of Annex VII to Regulation (EC) No 999/2001 refers to the methods and protocols set 
out in Annex X. The wording of this point should be amended to reflect amendments to Annex X brought by 
this act. 
(4)  Chapt er A of Annex VIII to Regulation (EC) No 999/2001 lays down rules governing intra-U nion trade in live 
animals, semen and embr yos, including the exemption of homozygous ovine ARR embr yos from any other clas­
sical scrapie related requirement in intra-Un ion trade. On 24 Januar y 2013, the European Food Safety Author ity 
(EFSA ) adopt ed a scientific opinion on the risk of transmission of classical scrapie via in vivo derived embr yo 
transfer in ovine animals (2), where it concluded that the risk of transmitting classical scrapie by the implantation 
of homozygous or heterozy gous ovine ARR embr yos could be considered negligible providing that the OIE 
recommendations and procedures relating to embr yo transfer are adhered to. The relevant provisions of Annex 
VIII should theref ore be amended to also exem pt intra-Union trade of hete rozygous ovine ARR embr yos from 
any other classical scrapie related requirement. 
(5)  In certain languag e versions of Regulation (EC) No 999/2001, there is a term inology incoherence between 
Points 1.2. and 1.3 of Section A of Chapt er A of Annex VIII to that Regulation and the rest of the text. For the 
sake of coherence, the same term should be used in the language versions concer ned. 
(6)  Point 2 of Section A of Chapt er A of Annex VIII to Regulation (EC) No 999/2001 lays down the rules governing 
the appro val of the negligible risk status for classical scrapie of a Member State or zone of a Member State. On 
4 July 2013, Austr ia submitted to the Commission the appropr iate suppor ting documentation. Given the favour­
able outcome of the assessment of this application by the Commission, Austr ia should be listed as a Member 
State with a negligible risk of classical scrapie. 
(7)  Point 3.2 of Section A of Chapt er A of Annex VIII to Regulation (EC) No 999/2001 lists Member State s with an 
appro ved national control programme for classical scrapie. Consider ing that Austr ia should be liste d as a Member 
State with a negligible risk of classical scrapie, it should simultaneously be delet ed from the list of Member States 
with an appro ved national control programme for classical scrapie, as this status offers guarant ees over and 
above those offered in the control programme. 29.10.2014 L 308/66 Official Jour nal of the European Union EN     
(1)OJ L 147, 31.5.2001, p. 1. 
(2)EFSA Jour nal 2013; 11(2):3080. (8)  Chapt er H of Annex IX to Regulation (EC) No 999/2001 lays down rules for the impor t in the Union of ovine 
and capr ine semen and embr yos. These imp ort rules should be update d to reflect amendments to Annex VIII 
brought by this act. 
(9)  Annex X to Regulation (EC) No 999/2001 lays down the methods of analysis applicable to TSE testing in bovine, 
ovine and capr ine animals. This Annex should be reviewed to updat e the information on the designated labora ­
tories, adjust the reference to various guidelines, harmonize some technical terms, and clarify the discr iminato ry 
testing process in case of positive TSE cases in ovine and capr ine animals, in accordance with the lates t scientifi c 
kno wledge and current practices in the Union. 
(10)  Point 4 of Chapt er C of Annex X to Regulation (EC) No 999/2001 sets out the lists of rapid tests appro ved for 
the monitori ng of TSEs in bovine, ovine and capr ine animals. On 18 Sept ember 2013, IDEXX made an applica ­
tion in order that the name of the test IDEXX HerdChek BSE-Scrapie Antigen Test Kit, EIA be chang ed to Herd ­
Chek BSE-Scrapie Antigen (IDEXX Laboratori es). The new packag e inser t for this test has been appro ved by the 
European Union Reference laboratory for TSEs on 2 May 2013. Further more, on 6 December 2013, the Enfe r 
Group informed that it has ceased the manufa cture of the Enfe r Version 3 TSE diagnostic kit and request ed the 
deletion of this kit from the list of appro ved BSE rapid tests in bovine animals. The lists in point 4 of Chapt er C 
of Annex X should theref ore be adapt ed according ly. 
(11)  In order to offer suffic ient time to Member States to align their scrapie-related certification procedures for ovine 
embr yos, certain amendments introduced by this Regulation should apply from 1 Januar y 2015. 
(12)  Regulation (EC) No 999/2001 should theref ore be amended according ly. 
(13)  The measures provided for in this Regulation are in accordance with the opinion of the Standing Committ ee on 
the Food Chain and Animal Health, 
HAS ADOPTED THIS REGUL ATION: 
Article 1 
Annex es II, VII, VIII, IX and X to Regulation (EC) No 999/2001 are amended in accordance with the Annex to this Regu ­
lation. 
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. 
Paragraphs (a), (b) and (e) of point 3 and point 4 of the Annex shall apply from 1 Januar y 2015. 
This Regulation shall be binding in its entirety and directly applicable in all Member State s. 
Done at Brussels, 28 October 2014. 
For the Commission 
The Presid ent 
José Manuel BARR OSO  29.10.2014 L 308/67 Official Jour nal of the European Union EN     ANNEX 
Annex es II, VII, VIII, IX and X to Regulation (EC) No 999/2001 are amended as follows: 
(1)  Annex II is amended as follows: 
(a)  Points 1 and 2 of Chapt er B are replaced by the following: 
‘1.  Structure of the risk analysis 
The risk analyses shall compris e an entr y assessment and an exposure assessment. 
2.  Entr y assessment (exter nal challenge) 
2.1.  The entr y assessment shall consist of assessing the likelihood that the BSE agent has either been introduced 
into the countr y or region via commodities potentially contaminated with a BSE agent, or is already 
present in the countr y or region. 
The following risk factors shall be taken into account: 
(a) the presence or absence of the BSE agent in the countr y or region and, if the agent is present, its preva ­
lence based on the outcome of surveillance activities; 
(b)  the production of meat-and-bone meal or greav es from the BSE indigenous ruminant population; 
(c)  imported meat-and-bone meal or greav es; 
(d)  imported bovine and ovine and capr ine animals; 
(e)  imported animal feed and feed ingredients; 
(f)  imported products of ruminant origin for human consump tion, whic h may have contained tissues 
listed in point 1 of Annex V and may have been fed to bovine animals; 
(g)  imported products of ruminant origin for in vivo use in bovine animals. 
2.2.  Special eradication schem es, surveillance and other epidemiological investigations (especially surveillance 
for BSE conduct ed on the bovine animals population) relevant to the risk factors listed in point 2.1 should 
be taken into ac-count in carrying out the entr y assessment.’ 
(b)  In point 3 of Chapt er D, Table 2 is replaced by the following: 
‘Table 2 
Points targets for different adult bovine animals population sizes in a countr y or region 
Points targets for countr y or region 
Adult bovine animals population size 
(24 months and older) Type A surveillance Type B surveillance 
> 1 000 000 300 000 150 000 
900 001-1 000 000 214 600 107 300 
800 001-900 000 190 700 95 350 
700 001-800 000 166 900 83 450 
600 001-700 000 143 000 71 500 29.10.2014 L 308/68 Official Jour nal of the European Union EN     Points targets for countr y or region 
Adult bovine animals population size 
(24 months and older) Type A surveillance Type B surveillance 
500 001-600 000 119 200 59 600 
400 001-500 000 95 400 47 700 
300 001-400 000 71 500 35 750 
200 001-300 000 47 700 23 850 
100 001-200 000 22 100 11 500 
90 001-100 000 19 900 9 950 
80 001-90 000 17 700 8 850 
70 001-80 000 15 500 7 750 
60 001-70 000 13 000 6 650 
50 001-60 000 11 000 5 500 
40 001-50 000 8 800 4 400 
30 001-40 000 6 600 3 300 
20 001-30 000 4 400 2 200 
10 001-20 000 2 100 1 050 
9 001-10 000 1 900 950 
8 001-9 000 1 600 800 
7 001-8 000 1 400 700 
6 001-7 000 1 200 600 
5 001-6 000 1 000 500 
4 001-5 000 800 400 
3 001-4 000 600 300 
2 001-3 000 400 200 
1 001-2 000 200 100’  
(2)  In Annex VII, the first paragraph in point 2.2.1 of Chapt er B is replaced by the following: 
‘If BSE cannot be excluded after the results of the secondar y molecular testing carried out in accordance with the 
methods and protocols set out in Annex X, Chapt er C, point 3.2(c) (ii), the killing and compl ete destr uction, without 
dela y, of all animals, embr yos and ova identified by the inquir y referred to in the second to fifth indents of 
point 1(b).’ 29.10.2014 L 308/69 Official Jour nal of the European Union EN     (3)  In Annex VIII, Section A of Chapt er A is amended as follows: 
(a)  In point 1.2, paragraph (g) is replaced by the following: 
‘(g)  only the following ovine and capr ine embr yos/o va may be introduced: 
(i)  embr yos/o va from donor animals whic h have been kept since birth in a Member State with a negligible 
risk of classical scrapie, or in a holding with a negligible or a controlled risk of classical scrapie, or 
which meet the following requirements: 
—  they are permanently identified to enable trace back to their holding of birth 
—  they have been kept since birth in holdings in which no case of classical scrapie has been confir med 
during their residency 
—  they showed no clinical sign of classical scrapie at the time of embr yo/ova collection; 
(ii)  ovine embr yos/o va carrying at least one ARR allele.’ 
(b)  In point 1.3, paragraph (g) is replaced by the following: 
‘(g)  only the following ovine and capr ine embr yos/o va may be introduced: 
(i)  embr yos/o va from donor animals whic h have been kept since birth in a Member State with a negligible 
risk of classical scrapie, or in a holding with a negligible or a controlled risk of classical scrapie, or 
which meet the following requirements: 
—  they are permanently identified to enable trace back to their holding of birth 
—  they have been kept since birth in holdings in which no case of classical scrapie has been confir med 
during their residency 
—  they showed no clinical sign of classical scrapie at the time of embr yo/ova collection; 
(ii)  ovine embr yos/o va carrying at least one ARR allele.’ 
(c)  In point 2, the following point 3 is added: 
‘2.3.  The Member States or zone of the Member State with a negligible risk for classical scrapie are the 
following: 
—  Austr ia.’ 
(d)  Point 3.2 is replaced by the following: 
‘3.2.  The national scrapie control programmes of the following Member States are hereb y appro ved: 
—  Denmark 
—  Finland 
—  Sweden.’ 
(e)  In point 4.2, paragraph (e) is replaced by the following: 
‘(e)  in the case of ovine embr yos, be carrying at least one ARR allele.’ 
(4)  In Annex IX, point (ii) of point 2 of Chapt er H is replaced by the following: 
‘(ii)  in the case of ovine embr yos, the embr yos carry at least one ARR allele.’ 29.10.2014 L 308/70 Official Jour nal of the European Union EN     (5)  Annex X is replaced by the following: 
‘ANNEX X 
REFERENCE LABORA TORIES, SAM PLING AND LABORA TOR Y ANAL YSIS METHODS 
CHA PTER A 
National referenc e laborator ies 
1.  The designat ed national reference laborat ory is to: 
(a)  have at its disposal facilities and exper t personnel enabling it to show at all times, and especially when the 
disease in question first appears, the type and strain of the agent of TSE, and to confir m results obtained by 
official diagnostic laboratories. Where it is not capable of identifying the strain-type of the agent, it shall set 
up a procedure to ensure that the identif ication of the strain is refer red to the EU reference laborat ory; 
(b)  verify diagnostic methods used in official diagnostic laboratories; 
(c)  be responsible for coordination of diagnostic standards and methods within the Member State. To this end, it: 
—  may provide diagnostic reag ents to official diagnostic laboratori es; 
—  is to control the quality of all diagnostic reagents used in the Member State 
—  is to periodically arrange comparative tests 
—  is to hold isolat es of the agents of the disease in question, or corresponding tissues containing such 
agents, coming from cases confir med in the Member State 
—  is to ensure conf irmation of results obtained in diagnostic laboratori es; 
(d)  is to cooperate with the EU reference laboratory , which includes the participation in the periodic compar ative 
tests orga nised by the EU referen ce laborat ory. Should a national reference laboratory fail in a comparat ive 
test organised by the EU reference laborat ory, it shall take immediate ly all the corrective actions to remedy 
the situation and successfully pass the repeat compar ative test or the next compar ative test organised by the 
EU reference laborat ory. 
2.  However , by way of derogation from point 1, Member States which do not have a national reference laborat ory 
shall use the services of the EU reference laboratory or of national referen ce laboratories locat ed in other Member 
States or European Free Trade Association (EFT A) Members. 
3.  The national reference laborat ories are: 
Austr ia: Agen tur für Gesundheit und Ernähr ungssic herheit GmbH (AGES) Institut für veteri när­
medizinische Untersuchung en 
Rober t Koch Gasse 17 
A-2340 Mödling 
Belgium: CER VA-COD A-VAR 
Centre d'Étude et de Rech erches Vétérinaires et Agrochimiques, Centr um voor Onder ­
zoek in Diergeneesk unde en Agroche mie, Veterinar y and Agroch emical Research Centre 
Groeselenberg 99 
B-1180 Bruxelles 
Bulgar ia: Национален диагностичен научноизследователски ветеринарномедицински институт 
‘Проф. Д-р Георги Павлов’ 
Национална референтна лаборатория ‘Tрансмисивни спонгиформни енцефалопатии’ 
бул. ‘Пенчо Славейков’ 15 
София 1606 
(National Diagnostic Veterinar y Research Institute “Prof. Dr Georgi Pavlov”, National 
Reference Laboratory for Transmissible Spongif orm Encephalopathies, 15 Penc ho 
Slave ykov Blvd., 1606 Sofia) 29.10.2014 L 308/71 Official Jour nal of the European Union EN     Croatia: Hrvatski vete rinarski institut, 
Savska Cesta 143 
10000 Zagreb 
Cypr us: State Veterinar y Laborator ies 
Veterinar y Services 
CY-1417 Athal assa 
Nicosia 
Czech Republic: Státní veteri nární ústav Jihla va (State Veterinar y Institute Jihla va) 
National Reference Laboratory for BSE and Animal TSEs 
Rantířo vská 93 
586 05 Jihla va 
Denmark: Veterinær institutte t 
Danmark s Tekniske Universitet 
Bülo wsvej 27 
DK-1870 Freder iksbe rg C 
(National Veterinar y Institute, Techn ical Universit y of Denmark, 27, Bülo wsvej, DK — 
1870 Freder iksberg C) 
Estonia: Veterinaar - ja Toidulaboratoori um (Estonian Veterinar y and Food Laboratory) 
Kreutzwaldi 30 
Tartu 51006 
Finland: Finnish Food Safety Author ity Evira 
Research and Laborat ory Depar tment 
Veterinar y Virology Research Unit- TSEs 
Mustialankatu 3 
FI-00790 Helsinki 
France: ANSES-L yon, Unité MND 
31, avenue Tony Garnier 
69 364 LYON Cedex 07 
German y: Friedr ich-Loeff ler-Institut 
Institute for Novel and Emerging Infectious Diseases at the Frieder ich-Loeff ler-Institut 
Federal Research Institute for Animal Health 
Suedufer 10 
D-17493 Greifswald Insel Riems 
Greece: Ministr y of Agriculture — Veterinar y Laborat ory of Larissa 
6th km of Larissa — Trikala Highwa y 
GR-41110 Larissa 
Hungar y: Veterinar y Diagnostic Directorat e, National Food Chain Safety Office (VDD NFCSO) 
Tábor nok u. 2 
1143 Budapest 
Ireland: Central Veterinar y Research Laboratory 
Depar tment of Agriculture, Food and the Mar ine 
Backwest on Cam pus 
Celbr idge 
Co. Kildare 29.10.2014 L 308/72 Official Jour nal of the European Union EN     Italy : Istituto Zooprofilattico Sper imentale del Piemonte , Ligur ia e Valle d'Aosta — CEA 
Via Bologna, 148 
I-10154 Torino 
Latvia: Institute of Food Safety , Animal Health and Environment (BIOR) 
Lejupes Str. 3 
Riga LV 1076 
Lithuania: National Food and Veterinar y Risk Assessment Institute 
J. Kairiūkščio str. 10 
LT-08409 Vilnius 
Luxembourg: CER VA-COD A-VAR 
Centre d'Étude et de Rech erches Vétérinaires et Agrochimiques, Centr um voor Onder ­
zoek in Diergeneesk unde en Agroche mie, Veterinar y and Agroch emical Research Centre 
Groeselenberg 99 
B-1180 Bruxelles 
Malta: Veterinar y Diagnostic Laboratory 
Depar tment of Food Health and Diagnostics 
Veterinar y Affairs and Fisher ies Division 
Ministr y for Rural Affairs and the Environment 
Alber t Town Marsa 
Netherlands: Central Veterinar y Instutute of Wageningen UR 
Edelher tweg 15 
8219 PH Lelystad 
P.O. Box 2004 
NL-8203 AA Lelystad 
Poland: Państwo wy Instytut Weterynar yjny (PIW et) 
24-100 Puławy 
al. Partyzant ów 57 
Portugal: Setor diagnóstico EET 
Laboratóri o de Patolo gia 
Unidade Estratégica de Investi gação e Serviços de Produção e Saúde Animal 
Instituto Nacional de Investi gação Agrár ia e Veterinár ia Rua General Morais Sarmento 
1500-311 Lisboa 
Romania: Institutul de Diagnostic și Sănătate Animală (Institute for Diagnosis and Animal Health) 
Depar tment of Mor phology 
Strada Dr Staico vici nr. 63, 5 
București 050557 
Slovakia: State Veterinar y Institute Zvolen 
Pod dráhami 918 
SK-960 86, Zvolen 
Slovenia: Universit y of Ljubljana, Veterinar y faculty 
National Veterinar y Institute 
Gerbičeva 60 
SI-1000 Ljubljana 29.10.2014 L 308/73 Official Jour nal of the European Union EN     Spain: Laboratori o Central de Veterinar ia (Alg ete) 
Ctra. M-106 pk 1,4 
28110 Alget e (Madr id) 
Sweden: National Veterinar y Institute 
S-751 89 Uppsala 
Unit ed Kingdom: Animal Health and Veterinar y Laboratories Age ncy 
Woodham Lane 
New Haw, Addlestone, Surrey KT15 3NB  
CHA PTER B 
EU reference laboratory 
1.  The EU referen ce laboratory for TSEs is: 
The Animal Health and Veterinar y Laboratori es Agency 
Woodham Lane 
New Haw 
Addlestone 
Surrey KT15 3NB 
Unit ed Kingdom 
2.  The functions and duties of the EU reference laboratory are: 
(a) to coordinate, in consultation with the Commission, the methods employed in the Member States for diag ­
nosing TSEs and the deter mination of the prion prote in genotype in ovine animals, specifically by: 
—stori ng and supplying corresponding tissues containing the TSE agents, for the development or produc ­
tion of the relevant diagnostic tests or for typing strains of the TSE agents 
—  supplying standard sera and other reference reage nts to the national referen ce laboratori es in order to 
standardise the tests and reage nts used in the Member States 
—  building up and retaining a collection of corresponding tissues containing the agents and strains of TSEs 
—organising periodic comparative tests for the procedures for the diagnosis of TSEs and for the deter min ­
ation of the prion prot ein genotype in ovine animals at EU level 
—  collecting and collating data and information on the methods of diagnosis used and the results of tests 
carried out in the EU 
—  character ising isolate s of the TSE agent by the most up-to -date methods to allow greate r understanding of 
the epidemiology of the disease 
—  keeping abreast of trends in surveillance, epidemiology and prevention of TSEs throughout the world 
—  maintaining exper tise on prion diseases to enable rapid differ ential diagnosis 
—  acquir ing a thorough kno wledge of the preparation and use of diagnostic methods used to control and 
eradicate TSEs; 
(b)  to assist actively in the diagnosis of outbreaks of TSEs in Member States by studying samples from TSE- 
infect ed animals sent for conf irmatory diagnosis, characteri sation and epidemiological studies; 
(c)  to facilitate the training or retraining of exper ts in laborat ory diagnosis with a view to the harmonisation of 
diagnostic techniques throughout the EU. 29.10.2014 L 308/74 Official Jour nal of the European Union EN     CHAPTER C 
Sampling and laborato ry testing 
1.  Sampling 
Any samples intended to be examined for the presence of a TSE shall be collect ed using the methods and prot o­
cols laid down in the latest edition of the Manual for diagnostic tests and vaccines for Terrestr ial Animals of the 
World Org anisation for Animal Health (OIE) (the Manual). In addition to, or in the absence of, OIE methods 
and protocols, and to ensure that suffi cient mater ial is available, the compet ent author ity shall ensure the use of 
sampli ng methods and protocols in accordance with guidelines issued by the EU reference laborat ory. 
In particular the compet ent author ity shall collect the appropr iate tissues, according to the available scientific 
advice and the guidelines of the EU reference laborat ory, in order to ensure the detection of all kno wn strains 
of TSE in small ruminants and shall keep at least half of the collect ed tissues fresh but not frozen until the 
result of the rapid test is negat ive. Where the result is positive or inconclusive the residual tissues must be 
subject to conf irmatory testing, and be processed subsequently in accordance with the EU reference laboratory 
guidelines on discr iminato ry testin g and classification — “TSE strain charact erisation in small ruminants: A 
technical handbook for National Reference Laborat ories in the EU”. 
The samp les shall be correctly mark ed as to the identity of the samp led animal. 
2.  Laborator ies 
Any laborat ory examination for TSE shall be carried out in official diagnostic laborat ories designat ed for that 
purpose by the compet ent author ity. 
3.  Methods and protocols 
3.1.  Labor atory testing for the presence of BSE in bovine animals 
(a)  Suspect cases 
Sample s from bovine animals sent for laborat ory testing pursuant to the provisions of Article 12(2) shall 
immediately be subjected to conf irmatory examinations using at least one of the following methods and 
protocols laid down in the latest edition of the Manual: 
(i)  the immunohistoc hemical (IHC) method; 
(ii)  Wester n blot ; 
(iii)  the demonstration of charact eristic fibrils by electron microscop y; 
(iv)  histo pathological examination; 
(v)  the combination of rapid tests as laid down in the third subparagraph. 
If the histopathological examination is inconclusive or negative, the tissues shall be submitt ed to a further 
examination by one of the other conf irmat ory methods and protocols. 
Rapid tests may be used for both primar y screening of suspect cases and, if inconclusive or positive, for 
subsequent conf irmation, according to the guidelines from the EU reference laboratory — “OIE rules for 
the official conf irmation of BSE in bovines (based on an initial reactive result in an appro ved rapid test) by 
using a second rapid test”, and provided that : 
(i)  the confir mation is carried out in a national reference laboratory for TSEs; and 
(ii)  one of the two rapid tests is a Wester n blot ; and 29.10.2014 L 308/75 Official Jour nal of the European Union EN     (iii)  the second rapid test used: 
—  includes a negative tissue control and a bovine BSE samp le as positive tissue control, 
—  is of a different type than the test used for the primar y screening; and 
(iv)  if a rapid Western blot is used as the first test, the result of that test must be documented and the blot 
image submitted to the national reference laboratory for TSEs; and 
(v)  where the result of the primar y screening is not conf irmed by the subsequent rapid test, the sample 
must be subjected to an examination by one of the other conf irmatory methods; where the histopatho ­
logical 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 conf irmat ory methods and protocols. 
If the result of one of the conf irmatory examinations refer red to in points (i) to (v) of the first sub­
paragraph is positive, the animal shall be rega rded as a positive BSE case. 
(b)  BSE monitor ing 
Sample s from bovine animals sent for laborat ory testing pursuant to the provisions of Annex III, Chapt er 
A, Part I shall be examined by a rapid test. 
When the result of the rapid test is inconclusive or positive, the sam ple shall immediately be subjecte d to 
conf irmatory examinations using at least one of the following methods and protocols laid down in the 
latest edition of the Manual: 
(i)  the immunohistoc hemical (IHC) method; 
(ii)  Wester n blot ; 
(iii)  the demonstration of charact eristic fibrils by electron microscop y; 
(iv)  histo pathological examination; 
(v)  the combination of rapid tests as laid down in the fourth subparagraph. 
Where the histo pathological examination is inconclusive or negative, the tissues shall be submitted to a 
further examination by one of the other conf irmat ory methods and protocols. 
Rapid tests may be used for both primar y screening and, if inconclusive or positive, for subsequent conf ir­
mation, according to the guidelines from the EU reference laboratory — “OIE rules for the official conf ir­
mation of BSE in bovines (based on an initial reactive result in an appro ved rapid test) by using a second 
rapid test”, and provided that: 
(i)  the confir mation is carried out in a national reference laboratory for TSEs; and 
(ii)  one of the two rapid tests is a Wester n blot ; and 
(iii)  the second rapid test used: 
—  includes a negative tissue control and a bovine BSE samp le as positive tissue control, 
—  is of a different type than the test used for the primar y screening; and 
(iv)  if a rapid Western blot is used as the first test, the result of that test must be documented and the blot 
image submitted to the national reference laboratory for TSEs; and 
(v)  where the result of the primar y screening is not conf irmed by the subsequent rapid test, the sample 
must be subjected to an examination by one of the other conf irmatory methods; where the histopatho ­
logical 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 conf irmat ory methods and protocols. 29.10.2014 L 308/76 Official Jour nal of the European Union EN     An animal shall be regar ded a positive BSE case if the result of the rapid test is inconclusive or positive, 
and at least one of the conf irmat ory examinations refer red to in points (i) to (v) of the second sub­
paragraph is positive. 
(c)  Fur ther examination of positive BSE cases 
Sample s from all positive BSE cases shall be forwarded to a laboratory , appoint ed by the compet ent 
author ity, which has participat ed successfully in the latest prof iciency testing organised by the EU reference 
laboratory for discr iminato ry testing of conf irmed BSE cases, where they shall be further tested in accord ­
ance with the methods and protocols laid down in the EU reference laboratory' s method for the classifica ­
tion of bovine TSE isolat es (a two- blot method for the provisional classification of bovine TSE isolates). 
3.2.  Labor atory testing for the presence of TSE in ovine and capr ine animals 
(a)  Suspect cases 
Sample s from ovine and capr ine animals sent for laboratory testing pursuant to the provisions of 
Article 12(2) shall immediate ly be subjected to conf irmat ory examinations using at least one of the 
following methods and protocols laid down in the latest edition of the Manual: 
(i)  the immunohistoc hemical (IHC) method; 
(ii)  Wester n blot ; 
(iii)  the demonstration of charact eristic fibrils by electron microscop y; 
(iv)  histo pathological examination. 
In case the histo pathological examination is inconclusive or negative, the tissues shall be submitted to a 
further examination by one of the other conf irmat ory methods and protocols. 
Rapid tests may be used for primar y screening of suspect cases. Such tests may not be used for subsequent 
conf irmation. 
Where the result of the rapid test used for primar y screening of suspect cases is positive or inconclusive, 
the samp le shall be subjected to an examination by one of the conf irmatory examinations refer red to in 
points (i) to (iv) of the first subparagraph. Where the histopathological examination is used for that 
purpose, but proves to be inconclusive or negative, the tissues shall be submitte d to a further examination 
by one of the other conf irmat ory methods and protocols. 
If the result of one of the confir matory examinations refer red to in points (i) to (iv) of the first sub­
paragraph is positive, the animal shall be regarded as a positive TSE case and further examination as 
refer red to in point (c) shall be performe d. 
(b)  TSE monit oring 
Sample s from ovine and capr ine animals sent for laboratory testing pursuant to the provisions of Annex 
III, Chapt er A, Part II (Monit oring in ovine and capr ine animals) shall be examined by a rapid test, in order 
to ensure the detec tion of all kno wn strains of TSE. 
When the result of the rapid test is inconclusive or positive, the sampled tissues shall immediately be sent 
to an official laboratory for conf irmatory examinations by histo pathology , immunohist ochemistry , Western 
blotting or demonstration of charact eristic fibrils by electron microscop y, as refer red to in point (a). If the 
result of the conf irmatory examination is negat ive or inconclusive, the tissues shall be submitte d to a 
further examination by immunohistoc hemistr y or Western blotting. 
If the result of one of the confir matory examinations is positive, the animal shall be regar ded as a positive 
TSE case and further examination as refer red to in point (c) shall be performe d. 29.10.2014 L 308/77 Official Jour nal of the European Union EN     (c)  Fur ther examination of positive TSE cases 
(i)  Primar y molecular testing with a discr iminato ry Western blotting method 
Samples from clinical suspect cases and from animals tested in accordance with Annex III, Chapt er A, 
Part II, points 2 and 3 which are regar ded as positive TSE cases but which are not atypical scrapie 
cases following the examinations referred to in points (a) or (b), or which displa y charact eristics 
which are deemed by the testing laboratory to mer it inve stigation, shall be examined using a 
discr iminator y Western blotting method listed in the guidelines of the EU reference laboratory by an 
official diagnostic laboratory designat ed by the compet ent author ity, which has participated success ­
fully in the latest proficiency testin g organised by the EU reference laboratory for the use of such a 
method. 
(ii)  Secondar y molecular testing with additional molecular testing methods 
TSE cases in which the presence of BSE cannot be excl uded according to the guidelines issued by the 
EU reference laboratory by the primar y molecular testing refer red to in point (i), shall be refer red 
immediately to the EU reference laborat ory, with all the relevant information available. The samples 
shall be submitte d to further inve stigation and conf irmation by at least one altern ative method, 
differi ng immunochem ically from the original primar y molecular method, depending on the volume 
and nature of the refer red mater ial, as descr ibed in the guidelines of the EU reference laborat ory. 
These additional tests will be carried out in the following laborat ories appro ved for the relevant 
method: 
Agence Nationale de Sécur ité Sanitaire de l'alimentation, de l'env ironnement et du travail 
31, avenue Tony Garnier 
BP 7033 
F-69342 Lyon Cedex 
Commissar iat à l'Energie Atomique 
18, route du Panorama 
BP 6 
F-92265 Font enay-aux-R oses Cedex 
Animal Health and Veterinar y Laboratories Agen cy 
Woodham Lane 
New Haw 
Addlest one 
Surrey KT15 3NB 
Unit ed Kingdom 
The results shall be interpr eted by the EU referen ce laboratory assisted by a panel of exper ts refer red 
to as the Strain Typing Exper t Group (STEG), including a representative of the relevant national refer­
ence laborat ory. The Commission shall be informed immediately about the outcome of that interpre­
tation. 
(iii)  Mouse bioassa y 
Samples indicative of BSE or inconclusive for BSE, following secondar y molecular testing, shall be 
further analysed by mouse bioassa y for final conf irmation. The nature or quantity of available mate ­
rial may influence the bioassa y design, which will be appro ved by the EU reference laboratory assisted 
by the STEG on a case by case basis. Bioassa ys will be performed by the EU reference laboratory , or 
by laborat ories designated by the EU reference laborat ory. 
The results shall be interpret ed by the EU reference laborat ory assiste d by the STEG. The Commission 
shall be informed immediately about the outcome of that interpretat ion. 
3.3.  Labor atory testing for the presence of TSEs in species other than those referred to in points 3.1 and 3.2 
Where methods and prot ocols are established for tests carried out to conf irm the suspecte d presence of a TSE 
in a species other than bovine, ovine and capr ine, they shall include at least a histopathological examination of 
brain tissue. The compet ent author ity may also require laborat ory tests such as immunohistoc hemistr y, Western 
blotting, demonstration of charact eristic fibrils by electron microscop y or other methods designed to detect the 
disease associated form of the prion prote in. In any case at least one other laborat ory examination shall be 
carried out if the initial histopathological examination is negativ e or inconclusive. At least three diffe rent exami ­
nations with positive results shall be carried out in the event of the first appearance of the disease. 29.10.2014 L 308/78 Official Jour nal of the European Union EN     In particular , where BSE is suspect ed in a species other than bovine animals, the cases shall be refer red to the 
EU reference laboratory assisted by the STEG for further character isation. 
4.  Rapid tests 
For the purposes of carrying out the rapid tests in accordance with Articles 5(3) and 6(1), only the following 
methods shall be used as rapid tests for the monitori ng of BSE in bovine animals: 
—the immunoblotting test based on a Wester n blotting procedure for the detection of the Proteinase K-resis ­
tant fragment PrPRes (Prionics-Check Western test), 
—  the sandwich immunoassa y for PrPRes detection (shor t assa y protocol) carried out following denaturation 
and concentration steps (Bio-Rad TeSeE SAP rapid test), 
—  the microplate -based immunoassa y (ELISA) which detects Proteinase K-resistant PrPRes with monoclonal 
antibodies (Prionics-Check LIA test), 
—the immunoassa y using a chemical polymer for selective PrPSc captur e and a monoclonal detection anti­
body directe d against conser ved regions of the PrP molecule (IDEXX HerdChek BSE Antigen Test Kit, EIA & 
HerdChek BSE-Scrapie Antige n (IDEXX Laborat ories)), 
—  the later al-flow immunoassa y using two different monoclonal antibodies to detect Proteinase K-resistant PrP 
fractions (Prionics Check PrioSTRIP), 
—  the two- sided immunoassa y using two different monoclonal antibodies direct ed against two epitopes 
present ed in a highly unfolded state of bovine PrPSc (Roboscreen Beta Prion BSE EIA Test Kit). 
For the purposes of carrying out the rapid tests in accordance with Articles 5(3) and 6(1), only the following 
methods shall be used as rapid tests for the monitori ng of TSE in ovine and capr ine animals: 
—  the sandwich immunoassa y for PrPRes detection (shor t assa y protocol) carried out following denaturation 
and concentration steps (Bio-Rad TeSeE SAP rapid test), 
—  the sandwic h immunoassa y 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-R ad TeSeE 
Sheep/Goat rapid test), 
—the immunoassa y using a chemical polymer for selective PrPSc captur e and a monoclonal detection anti­
body direct ed against conser ved regions of the PrP molecule (HerdChek BSE-Scrapie Antigen (IDEXX 
Laboratories)), 
—  the later al-flow immunoassa y using two different monoclonal antibodies to detect Proteinase K-resistant PrP 
fractions (Prionics — Check PrioSTRIP SR, visual reading protocol) . 
In all rapid tests, sample tissue on whic h the test must be applied must comply with the manufacturer's instr uc­
tions for use. 
Producers of rapid tests must have a quality assurance system in place that has been appro ved by the EU refer­
ence laboratory and ensures that the test performa nce does not change. Producers must provide the EU refer ­
ence laboratory with the test protocols. 
Changes to rapid tests and to test protocols may only be made after prior notifi cation to the EU reference 
laboratory and provided that the EU referen ce laboratory finds that the change does not alter the sensitivity , 
specifi city or reliability of the rapid test. That finding shall be communicated to the Commission and to the 
national reference laboratori es. 
5.  Alter nativ e tests 
(To be defined)’  29.10.2014 L 308/79 Official Jour nal of the European Union EN