23.9. 83 Official Journal of the European Communities No C 255/3 
II 
(Preparatory Acts) 
COMMISSION 
Proposal for a Council Directive amending Directive 64/432/EEC as regards brucellosis 
in respect of the buffered brucella antigen test, the micro-agglutination test and the milk 
ring test as applied to samples of milk from bulk tanks 
(Submitted by the Commission to the Council on 9 September 1983) 
THE COUNCIL OF THE EUROPEAN 
COMMUNITIES, 
Having regard to the Treaty establishing the 
European Economic Community, and in particular 
Article 43 thereof, HAS ADOPTED THIS DIRECTIVE: 
Article 1 
Directive 64/432/EEC is hereby amended as follows: 
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-
Community trade in bovine animals and swine ('), as 
last amended by Directive 82/893/EEC (2), lays down 
common standards on anti-brucellosis measures 
applicable to animals intended for intra-Community 
trade; 
Whereas it is necessary to indicate precisely that the 
slow tube serum agglutination test is the test to be 
carried out on bovine animals and swine before they 
enter into intra-Community trade; 
Whereas, in order to assure continuous free trade 
within the Community of bovine animals, it is 
necessary to adapt the technical provisions of the 
above Directive relating to brucellosis to take account 
of scientific progress; 
Whereas new scientific knowledge and technical 
developments in the diagnosis and control of bovine 
brucellosis require adjustment of existing Community 
measures in this field, 
O OJ No 121, 29. 7. 1964, p. 1977/64. 
O OJ No L 378, 31. 12. 1982, p. 57. 1. In Article 3 (3), subparagraph (c), the words 
'paragraph A' are added after the words 'Annex 
C. 
2. In Article 3 (4), subparagraph (i), the words 
'paragraph A' are added after the words 'Annex 
C\ 
3. In Article 3 (6), subparagraph (c), the words 
'paragraph A' are added after the words 'Annex 
C. 
4. In Article 7 (1) C, the words 'paragraph A' are 
added after the words 'Annex C\ 
5. In Article 7 (1) D, the words 'paragraph A' are 
added after the words 'Annex C\ 
6. In Annex A.ILA.l (c) (i): 
— in the first paragraph the words 'paragraph A' 
are added after the words 'Annex C, 
— the second indent is replaced by the following: 
'— the sero-agglutination tests referred to in 
the first paragraph may be replaced by two 
official buffered brucella antigen tests or 
two micro-agglutination tests carried out 
in accordance with Annex C (D) and (G). 
These tests shall also be carried out at 
intervals of at least three months and at 
most 12 months.' 
No C 255/4 Official Journal of the European Communities 23. 9. 83 
7. Annex A.II.A.1 (c) (ii) is replaced by the following: 
'(ii) are checked annually to establish that 
brucellosis is not present by three ring tests 
carried out at intervals of at least three 
months or two ring tests 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) 
carried out at not less than six weeks after the 
second ring test. If ring tests are not carried 
out, two serological tests (sero-agglutination 
test or buffered brucella antigen test or 
plasma-agglutination test or plasma ring test 
or micro-agglutination test) shall be carried 
out each year at intervals of at least three 
months and not more than six months. 
Where, in a Member State or region thereof 
in which all bovine herds are subject to 
official operations to combat brucellosis, not 
more than 1 °/o of bovine herds are infected, it 
shall be sufficient to carry out each year two 
ring tests at an interval of at least three 
months, or one serological test (sero-aggluti­
nation test or buffered brucella antigen test or 
plasma-agglutination test or plasma ring test 
or micro-agglutination test).' 
In Annex A.II.A.2 (c), the last paragraph is 
replaced by the following: 
'The sero-agglutination tests referred to in 
1 (c) (i), first indent, may be replaced by buffered 
brucella antigen tests carried out in accordance 
with Annex C (D), or micro-agglutination tests 
carried out in accordance with Annex C (G).' 
9. In Annex C: 3. The antigen may be stained only with 
hematoxylin or tetrazolium; hema­
toxylin should preferably be used. 
4. If no preservation is used then the 
reaction test must be carried out 
between 18 and 24 hours of taking the 
sample from the cow. If milk is to be 
tested later than 24 hours after sampling 
then preservation must be used. 
Formalin or mercuric chloride may be 
used as preservatives and if either of 
these are used the test must be carried 
out within the following 14 days after 
the day of sampling. Formalin may be 
added to give a final dilution in the milk 
sample of 0,2 °/o and, in such cases, the 
ratio between the amount of milk and 
the solution of formalin must be at least 
10 to 1. A solution of mercuric chloride 
may alternatively be added to give a 
final dilution in the milk of 0,2 % and, 
in such cases, the ratio between the 
amount of milk and the solution of 
mercuric chloride must be 10 to 1. 
5. The reaction must be carried out using 
one of the following methods: 
— on a column of milk at least 25 mm 
high and on a volume of milk of 1 
ml to which 0,03 ml of one of the 
standardized stained antigens has 
been added, 
— on a column of milk at least 25 mm 
high and on a volume of milk of 1 
ml to which 0,05 ml of one of the 
standardized stained antigens has 
been added, 
— on a volume of milk of 8 ml to 
which 0,08 ml of one of the stan­
dardized stained antigens has been 
added. 
paragraph C is replaced by the following: 
'C. Ring test 
1. The ring test must be made on the 
contents of each milk churn or on the 
contents of each bulk tank from the 
farm. 
2. The standard antigen to be used must 
come from one of the institutes listed in 
paragraph A.9 (a) to (j). It is 
recommended that the antigens should 
be standardized according to the 
WHO/FAO recommendations. 6. The mixture of milk and antigens must 
be incubated at 37 ° C for not less than 
45 minutes and not more than 60 
minutes. The test must be assessed 
within 15 minutes of removal from the 
incubator. 
7. The reaction must be assessed according 
to the following criteria: 
(a) negative reaction: coloured milk, 
colourless cream; 
(b) positive reaction: milk and cream 
identically colour­
ed or colourless 
milk and colour­
ed cream.' 
23.9.83 Official Journal of the European Communities No C 255/5 
a new paragraph G is added: 
'G. Micro-agglutination test 
1. Diluents are made up of 0,85 % 
physiological saline solution phenolized 
at 0,5 % phenol and the antigen itself is 
stained with safranin 0. 
2. The antigen shall be prepared as 
described under points 6, 7 and 8 of 
Annex C (A) and shall be titrated as 
described under point 5 of Annex C (A). 
3. The standard serum is the same as that 
under point 1 of Annex C (A). 
4. The standard serum must be supplied by 
the Bundesgesundheitsamt, Berlin. 
5. The micro-agglutination test shall be 
carried out on plates bearing wells and at least three dilutions must be prepared 
for each serum. Dilutions of suspect 
serum must be made in such a way that 
the reading of the reaction at the 
infection limit is made in the median 
well by comparison with positive and 
negative controls. If there is a positive 
reaction in the median well, the suspect 
serum contains at least 30 IU of aggluti­
nation per ml.' 
Article 2 
Member States shall bring into force the laws, regu­
lations and administrative provisions necessary to 
comply with this Directive by 1 July 1984. They shall 
forthwith inform the Commission thereof. 
Article 3 
This Directive is addressed to the Member States. 
Recommendation for a Council Regulation on the conclusion of a Protocol on financial 
and technical cooperation between the European Economic Community and the Republic 
of Cyprus 
(Submitted by the Commission to the Council on 8 September 1983) 
THE COUNCIL OF THE EUROPEAN 
COMMUNITIES, 
Having regard to the Treaty establishing the 
European Economic Community, and in particular 
Article 238 thereof, 
Having regard to the recommendation from the 
Commission, 
Having regard to the opinion of the European 
Parliament, 
Whereas the Protocol on financial and technical 
cooperation between the European Economic 
Community and the Republic of Cyprus, signed on 
..., should be approved, 
HAS ADOPTED THIS REGULATION: 
Article 1 
The Protocol on financial and technical cooperation 
between the European Economic Community and the Republic of Cyprus is hereby approved on behalf of 
the Community. 
The text of the Protocol is attached to this Regu­
lation. 
Article 2 
The President of the Council shall give the 
notification provided for in Article 21 (1) of the 
Protocol. 
Article 3 
This Regulation shall enter into force on the day 
following its publication in the Official Journal of the 
European Communities. 
This Regulation shall be binding in its entirety and 
directly applicable in all Member States.