PROPOSAL FOR A COUNCIL DIRECTIVE AMENDING DIRECTIVE 64/432/EEC AS REGARDS THE DIAGNOSIS OF BOVINE BRUCELLOSIS AND ENZOOTIC BOVINE LEUKOSIS
Date: 1990-11-09
Institutions: European Commission
Subjects: animal leucosis, brucellosis, cattle, medical diagnosis

IIP  COMMISSION OF THE EUROPEAN  COMMUNITIES 

mm 

HI 

COM(90)492  f inal 

Brussels, 9  November 1990 

Proposal  for a 

COUNCIL DIRECTIVE 

amending  Directive 64/432/EEC as regards the diagnosis of 

bovine  brucellosis and enzootic  bovine  leukosis 

(presented  by the Commission) 

-  2 -

EXPLANATORY  MEMORANDUM 

Recent  advances  in Scientific  knowledge  have allowed  for  the development  of 

new  tests  (ELISA)  for  the  diagnosis  and  control  of  bovine  brucellosis  and 

for  the adaptation of  tests for  the detection of enzootic bovine  leukosis. 

A scientific  group  (Brucellosis  uniformity  antigen  group)  have  studied  the 

matter  and  have  carried  out  a  number  of  international  scientific  trials, 

resulting  from which  recommedations have been made  to  Incorporate  the ELISA 

as  an  alternative  for  milk  tests  for  the  maintenance  of  officially 

brucellosis  free herds as well  as  individual  blood  samples. 

The attached  is designed  to accomplish  the abovementioned objectives. 

There are no financial  repercussions on  the Community  budget  involved. 

- 3 -

Proposal  for a 

COUNCIL  DIRECTIVE 

amending Directive 64/432/EEC as regards  the diagnosis of 

bovine brucellosis and enzootic bovine  leukosis 

THE COUNCIL  Of  THE EUROPEAN  COMMUNITIES, 

Having  regard  to  the  Treaty  establishing  the  European  Economic  Community, 

and  in particular Article 43, 

Having regard  to the proposal  from  the Commission, 

Having  regard  to the opinion of  the European  Parliament, 

Having  regard  to the opinion of  the Economic  and Social  Committee, 

Whereas  Council  Directive  64/432/EEC  of  26  June  1964  on  animal  health 

problems  affecting  Intra-Communlty  trade  In  bovine  animals  and  swine*1), 

as  last  amended  by  Directive  90/422/EEC*2),  lays  down  the  methods  for 

maintaining  the  status  of  officially  brucellosis  free  herds  and  enzootic 

bovine  leukosis  free herds; 

Whereas,  due  to new  scientific  knowledge  and  technical  developments  In the 

diagnosis  and  control  of  bovine  brucellosis  and  enzootic  bovine  leukosis, 

an  adjustment  of  existing  Community  measures  In  this  field  has  proved 

necessary; 

(1)  OJ No 121, 29.7.1964, p. 1977/64 

(2)  OJ No L 224, 18.8.1990, p. 9. 

HAS ADOPTED  THIS DIRECTIVE: 

-  4 -

Article 1 

The  Annexes  to  Directive  64/432/EEC  are  hereby  amended  in  accordance  with 

the Annex  to this  Directive. 

Article  2 

Member  States  shall  bring 

into  force  the 

laws,  regulations  and 

administrative provisions  necessary  to comply  with  this Directive  not  later 

than  1 January  1991. They  shall  forthwith  inform  the Commission  thereof. 

When  Member  States  adopt  these  provisions,  these  shall  contain  a  reference 

to  this  Directive  or  shall  be  accompanied  by  such  reference  at  the  time of 

their  official  publication.  The  procedure  for  such  reference  shall  be 

adopted  by Member  States. 

Article  3 

This Directive  is addressed  to the Member  States. 

Done at Brussels, 

For  the  CounciI, 

- 5 -

oHrifcÀ, 

t. Annex A,  11.A.1.(c)(ii) is replaced by the following: 

(il)  are  checked  annually  to  establish  that  brucellosis  Is  not 

present  by  three  ring  tests  or  milk  EL ISA  carried  out  at 

intervals  of  at  least  three months or  two  ring  tests or  milk 

EL ISA  at  an  interval  of  at  least  three  months  and  one 

serological  test  (sero-agglutination test or  buffered brucella 

antigen  test  or  plasma  agglutination  test  or  plasma  ring  test 

or micro-agglutination test or  individual  blood  ELISA) carried 

out  at  not  less  than  six  weeks  after  the  second  ring  test  or 

milk  ELISA.  If  ring  tests or  milk  ELISA  are  not  carried  out, 

two  serological  tests  (sero-agglutiantIon  test  or  buffered 

brucella  antigen  test  or  plasma  agglutination  test  or  plasma 

ring  test  or  micro-agglutination  test  or  individual  blood 

ELISA) shall  be carried out each year  at  Intervals of  at  least 

three months and not more  than six months. 

Where,  in  a  Member  States  or  region  thereof  in  which  all 

bovine  herds  are  subject  to  official  operations  to  combat 

brucellosis, not more  than 1% of bovine herds are  Infected, it 

shall  be  sufficient  to carry  out  each  year  two  ring  tests or 

two  milk  ELISA  at  an  Interval  of  at  least  threee  months, or 

one  serological  test  (sero-agglutination  test  or  buffered 

brucella  antigen  test or  plasma-agglutination  test  or  plasma-

ring  test  or  micro-agglutination  tests  or  individual  blood 

ELISA). 

Where  ring  tests are carried  out  on  bulk  tanks, the  number  of 

tests  referred  to  In  the  preceding  subparagraphs  shall  be 

doubled  and  the  intervals between  the tests shall  be halved." 

2. The following point  shall  be added  to Annex  C: 

"H. 

The ELISA  for brucellosis as described under  Annex  G" 

-  6 

3.  In Annex G: 

a) 

The following words shall  be added  to the title: 

"and brucellosls" 

b) 

Point C  is replaced by  the following: 

"C. 

Enzyme-linked  immunosorbent  assay  (ELISA)  for  enzootic 

bovine  leukosis and bovine brucellosis. 

1. 

For  the  ELISA  method,  the  materials  and  reagents 

to be used are as follows: 

(a)  solid  phase  microplates,  cuvettes  or  any 

other  sol id phase; 

(b)  the antigen  Is fixed  to the solid phase with 

or  without  the  aid  of  polyclonal  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  E.B.L..  The  control  antigen 

should  be  identical  to  the  antigen  except 

for  the  B.L.V.  antigens.  If  catching 

antibodies are coated  to the solid phase  the 

antibodies must  not  react  to other  antigens 

other  than B.L.V. or brucellosis antigens; 

(c) 

the biological  fluid  to be  tested; 

(d)  a  corresponding  positive  and  negative 

control ; 

(e)  conjugate; 

- 7 -

(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; 

(1)  a  reading  system 

appropriate  to  the 

substrate used. 

2. 

Standardization and sensitivity of  test: 

(a)  For enzootic bovine  leukosis  the  sensitivity 

of  the  EL I SA  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  E4  serum  will  be  supplied  by  the 

National  Veterinary Laboratory,  Copenhagen. 

(b) 

(i)  For brucellosis, bulk milk  samples  are 

classified  negative  if  they  give  a 

reaction  less  than  fifty  percent  of 

that  given by a 1  in 10 000 dilution 

-  8  -

of 

the 

second 

International 

Brucellosis  Standard  serum  made  up  in 

negatIve  milk; 

(II)  for 

brucellosis 

individual 

serum 

samples  are  classified  negative  if 

they  give  a  reaction  less  than  ten 

percent  of  that  given  by  a  1  in  200 

dilution  of  the  second  International 

Brucellosis  Standard  serum  made  up  in 

sal I ne  so I ut ion. 

The  brucellosis  ELISA  standards  shall 

be  as  specified  in  Annex  C,  A  1  and  2 

(to  be  used  at  the  dilutions  indicated 

on  the  label). 

3. 

Conditions  for  use  of  the  ELISA  test  for  E.B.L. 

The  ELISA  method  may  be  used  on  a  sample  of  milk 

or  whey  taken  from  the  ml Ik  collected  from  a  farm 

with  at  least  30% of  dairy  cows  in milk. 

If  use  is  made  of  one  of  these  abovement ioned 

possibilities,  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. 

If  one  of  the  samples  scores  positive,  the  herd 

must  remain  under  official  supervision  until  a 

negative  result  has  been  recorded  for  at  least  two 

individual 

tests  carried  out,  at  a  minimum 

Interval  of  four  months,  on  all  cattle  aged  more 

than 

six 

months, 

In 

accordance 

with 

the 

abovementioned  provisions  and  In a  laboratory  that 

is  directly  supervised  by  a  laboratory  mentioned 

in point  A." 

T»*k  * o r ce  PME 
T»*k  * o r ce  PME 

XOMMTITIVENgSS  AND  EMPLOYMENT  IMPACT  STATEMENT 
XOMMTITIVENgSS  AND  EMPLOYMENT  IMPACT  STATEMENT 

II 
ii 

— 

• • — y—  «•• 
• • — y— 

« •• 

i — — ^ —« 

i — — ^ —« 

I. 

what  ic the sj'.n reason  fur  Introducing the measure ? 

Tu use advanced technology to diaonose ch" *•»*** 

II. 

Features nf  i.K« businesses in Question. Ill particular : 
<*>  »rt there many  SMEs ? 

NO 

•<b) Are Uiey cnnt.irnirated  in region* which  at >•  -. 

i.  eligible tor regional airl in the Member atotcs ? 

H.  eligible under  ihp €*0F ? 

III. 

Whet direct obligations does this neesure t«pos« on business**  1 

-NONE 

IV. 

What  indirect obligation* are locel authorities  LikvLy to Impott  , • '!";1 
on businesses ? 

NONb 

n 

V. 

Arr Tti«r« any  special Measures in rrKpeec of SiMEs 7  Please specify. • -

WO 

•"'"';. 

Vi„ 

What is th*  ;.ik»ly effect on : 
(a) the competitiveness of businesses ? 

(b> «mployment 7 

NONE 

•. none 

VII. 

Have both  sides of industry been consulted 1 

Pl*<*i» indicate ir>e"»r opinions. 

m0t necessary 

ISSN 0254-1475 

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