Council Directive of 26 June 1990 on animal health conditions governing the movement and import from third countries of equidae (90/426/EEC) (repealed)

1. Introductory Text

2. CHAPTER 1 General provisions

   1. Article 1. This Directive lays down animal health conditions for the movement...

   2. Article 2. For the purposes of this Directive: ‘holding’ means an agricultural...

3. CHAPTER II Rules for the movement of equidae

   1. Article 3. Member States shall authorize the movement of equidae registered in...

   2. Article 4. (1) Equidae must show no clinical sign of disease at...

   3. Article 5. (1) A Member State which is not free of African...

   4. Article 6. Member States which implement an alternative control system providing guarantees...

   5. Article 7. (1) The equidae must be transported, as soon as possible,...

   6. Article 8. (1) Member States shall ensure that: registered equidae which leave...

   7. Article 9. The rules laid down in Council Directive 90/425/EEC of 26...

   8. Article 10. Veterinary experts from the Commission may, to the extent necessary...

4. CHAPTER III Rules for imports from third countries

   1. Article 11. (1) Equidae imported into the Community must satisfy the conditions...

   2. Article 12. (1) The importation of equidae into the Community shall only...

   3. Article 13. (1) The equidae must come from third countries:

   4. Article 14. Before the day of loading for transportation to the Member...

   5. Article 15. Importation of equidae from the territory of a third country...

   6. Article 16. (1) The equidae must be identified in accordance with Article...

   7. Article 17. Checks shall be carried out on the spot by veterinary...

   8. Article 18. (1) Immediately upon arrival in the Member State of destination,...

   9. Article 19. The Commission, acting in accordance with the procedure laid down...

   10. Article 20. . . . . . . . . . ....

   11. Article 21. . . . . . . . . . ....

5. CHAPTER IV Final provisions

   1. Article 22. The provisions of this Directive, and in particular those contained...

   2. Article 23. The Annexes to this Directive shall be amended by the...

   3. Article 24. (1) The Commission shall be assisted by the Standing Committee...

   4. Article 25. (1) The Commission shall be assisted by the Standing Committee...

   5. Article 26. Article 34 of Directive 72/462/EEC shall apply to the requirements...

   6. Article 27. Member States shall bring into force the laws, regulations and...

   7. Article 28. This Directive is addressed to the Member States.

6. 1. ANNEX A

      COMPULSORILY NOTIFIABLE DISEASES

      1. The following diseases are compulsorily notifiable:

      2. Dourine Glanders Equine encephalomyelitis (of all types, including VEE) Infectious...

   2. ANNEX B

   3. ANNEX C

   4. ANNEX D

      AFRICAN HORSE SICKNESS

      1. Reagents for the enzyme-linked immunosorbent assays (ELISA) described below may...

      2. 1. COMPETITIVE ELISA FOR THE DETECTION OF ANTIBODIES TO AFRICAN HORSE...

         1. 1.1. Test procedure

            1. 1.1.1. Preparation of plates

               1. 1.1.1.1. Coat ELISA plates with AHSV antigen extracted from infected cell...

               2. 1.1.1.2. Wash plates three times by flooding and emptying the wells...

            2. 1.1.2. Control wells

               1. 1.1.2.1. Titrate the positive control sera in a twofold dilution series,...

               2. 1.1.2.2. Add 50 μl of the negative control serum at a dilution...

               3. 1.1.2.3. Add 100 μl/well of blocking buffer to wells C and D...

               4. 1.1.2.4. Add 50 μl of blocking buffer to wells E, F, G...

            3. 1.1.3. Spot test method

               1. 1.1.3.1. Add a 1 in 5 dilution of each test serum...

            4. 1.1.4. Serum titration method

               1. 1.1.4.1. Prepare a twofold dilution series of each test sample (1...

            5. 1.1.5. Add 50 μl of guinea pig antisera, pre-diluted in blocking buffer,...

               1. 1.1.5.1. Incubate for 1 hour at 37 °C on an orbital shaker....

               2. 1.1.5.2. Wash plates three times and blot dry as before.

               3. 1.1.5.3. Add 50 μl of rabbit anti-guinea-pig horseradish peroxidase (HRP) conjugate pre-diluted...

               4. 1.1.5.4. Incubate for 1 hour at 37 °C on an orbital shaker....

               5. 1.1.5.5. Wash plates three times and blot dry as before.

            6. 1.1.6. Chromogen

            7. 1.1.7. Reading

         2. 1.2. Expression of results

            1. 1.2.1. Using a software package print out the optical density (OD)...

            2. 1.2.2. The diagnostic threshold (cut-off value) for test sera is 50 %...

      3. 2. INDIRECT ELISA FOR THE DETECTION OF ANTIBODIES TO AFRICAN HORSE...

         1. 2.1. Test procedure

            1. 2.1.1. Solid phase

               1. 2.1.1.1. ELISA plates are coated with recombinant AHSV-4 VP7 diluted in...

               2. 2.1.1.2. Wash the plates five times with distilled water containing 0,01 %...

               3. 2.1.1.3. Block the plates with phosphate buffered saline (PBS) + 5 % (w/v)...

               4. 2.1.1.4. Remove the blocking solution and gently tap the plates onto...

            2. 2.1.2. Test samples

               1. 2.1.2.1. Serum samples to be tested, and positive and negative control...

               2. 2.1.2.2. Wash the plates as described in step 2.1.1.2.

            3. 2.1.3. Conjugate

               1. 2.1.3.1. Dispense 100 μl/well of horseradish-peroxidase (HRP) -conjugated anti-horse gamma-globulin diluted in...

               2. 2.1.3.2. Wash the plates as described in step 2.1.1.2.

            4. 2.1.4. Cromogen/Substrate

               1. 2.1.4.1. Add 200 μl/well of chromogen/substrate solution (10 ml of 80,6 mM DMAB (dimethyl...

               2. 2.1.4.2. Read the plates at 600 nm (or 620 nm).

         2. 2.2. Interpretation of the results

            1. 2.2.1. Calculate the cut-off value by adding 0,6 to the value...

            2. 2.2.2. Test samples giving absorbance values lower than the cut-off are...

            3. 2.2.3. Test samples giving absorbance values greater than the cut-off + 0,15...

            4. 2.2.4. Test samples giving intermediate absorbance values are doubtful and a...

      4. 3. BLOCKING ELISA FOR THE DETECTION OF ANTIBODIES TO AFRICAN HORSE...

         1. 3.1. Test procedure

            1. 3.1.1. ELISA plates

               1. 3.1.1.1. Coat ELISA plates with recombinant AHSV-4 VP7 diluted in carbonate/bicarbonate...

               2. 3.1.1.2. Wash the plates five times with phosphate buffered saline (PBS)...

               3. 3.1.1.3. Stabilise the plate by treatment with a stabilising solution (in...

            2. 3.1.2. Test samples and controls

               1. 3.1.2.1. For screening: dilute test sera and controls 1 in 10...

               2. 3.1.2.2. For titration: prepare a twofold dilution series of test sera...

            3. 3.1.3. Conjugate

            4. 3.1.4. Wash the plates five times with PBST and blot dry...

            5. 3.1.5. Chromogen/Substrate

            6. 3.1.6. Reading

         2. 3.2. Interpretation of the results

            1. 3.2.1. Assay validation

            2. 3.2.2. Cut-off calculation

            3. 3.2.3. Interpretation of results