1. Introductory Text

  2. CHAPTER I GENERAL PROVISIONS

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

    2. Article 2.For the purposes of this Directive the following definitions shall...

  3. CHAPTER II RULES FOR THE MOVEMENT OF EQUIDAE BETWEEN MEMBER STATES

    1. Article 3.Member States shall authorise the movement of registered equidae 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 from African...

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

    5. Article 7.(1) Equidae must be transported, as soon as possible, from...

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

    7. Article 9.The rules laid down in Directive 90/425/EEC shall apply in...

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

  4. CHAPTER III RULES FOR IMPORTATION OF EQUIDAE FROM THIRD COUNTRIES

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

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

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

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

    8. Article 18.Checks shall be carried out on the spot by veterinary...

    9. Article 19.In accordance with the procedure referred to in Article 21(2):...

  5. CHAPTER IV FINAL PROVISIONS

    1. Article 20.Annexes I to IV shall be amended in accordance with...

    2. Article 21.(1) The Commission shall be assisted by the Standing Committee...

    3. Article 22.Directive 90/426/EEC, as amended by the acts listed in Annex...

    4. Article 23.This Directive shall enter into force on the 20th day...

    5. Article 24.This Directive is addressed to the Member States.

    1. ANNEX I

      COMPULSORILY NOTIFIABLE DISEASES

      1. The following diseases are compulsorily notifiable:

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

    2. ANNEX II

      MODEL

      1. Passport No …

      2. I, the undersigned, certify that the animal identified above meets...

      3. it has been examined today and shows no clinical sign...

    3. ANNEX III

      MODEL

    4. ANNEX IV

      AFRICAN HORSE SICKNESS

      1. PART A Serological tests

        1. 1. Indirect ELISA for the detection of antibodies to African horse...

          1. 1.1. Test procedure

            1. 1.1.1. Solid phase

              1. 1.1.1.1. Coat ELISA plates with recombinant AHSV-4 VP7 diluted in carbonate/bicarbonate...

              2. 1.1.1.2. Wash the plates five times with distilled water containing 0,01...

              3. 1.1.1.3. Block the plates with phosphate buffered saline (PBS) pH 7,2...

              4. 1.1.1.4. Remove the blocking solution and gently tap the plates onto...

            2. 1.1.2. Test samples

              1. 1.1.2.1. Serum samples to be tested, and positive and negative control...

              2. 1.1.2.2. Wash the plates five times with distilled water containing 0,01...

            3. 1.1.3. Conjugate

              1. 1.1.3.1. Dispense 100 μl/well of horseradish-peroxidase (HRP) -conjugated anti-horse gamma-globulin diluted...

              2. 1.1.3.2. Wash the plates five times with distilled water containing 0,01...

            4. 1.1.4. Chromogen/Substrate

              1. 1.1.4.1. Add 200 μl/well of chromogen/substrate solution (10 ml of 80,6...

              2. 1.1.4.2. Read the plates at 600 nm (or 620 nm).

          2. 1.2. Interpretation of the results

            1. 1.2.1. Calculate the cut-off value by adding 0,06 to the value...

            2. 1.2.2. Test samples giving absorbance values lower than the cut-off are...

            3. 1.2.3. Test samples giving absorbance values greater than the cut-off +...

            4. 1.2.4. Test samples giving intermediate absorbance values are considered to be...

        2. 2. Blocking 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. Coat ELISA plates with 50-100 ng of recombinant AHSV-4 VP7...

              2. 2.1.1.2. Wash the plates three times with phosphate buffered saline (PBS)...

            2. 2.1.2. Test samples and controls

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

              2. 2.1.2.2. Wash the plates five times with phosphate buffered saline (PBS)...

            3. 2.1.3. Conjugate

              1. 2.1.3.1. Dispense 100 μl/well of horseradish peroxidase-conjugated Mab anti-VP7. In advance,...

              2. 2.1.3.2. Wash the plates five times with phosphate buffered saline (PBS)...

            4. 2.1.4. Chromogen/Substrate

            5. 2.1.5. Reading

          2. 2.2. Interpretation of the results

            1. 2.2.1. Determine the blocking percentage (BP) of each sample by applying...

            2. 2.2.2. Samples showing a BP value higher than 50 % should...

            3. 2.2.3. Samples showing a BP value lower than 45 % should...

            4. 2.2.4. Samples showing a BP value between 45 % and 50...

      2. PART B Identification of the agent

        1. Real-time Reverse-Transcription Polymerase Chain Reaction (rRT-PCR)

          1. 1. Extraction of viral RNA

          2. 2. Real-time RT-PCR Procedure

            1. 2.1. Group-specific real-time RT-PCR by Agüero et al., 2008

            2. 2.2. Group-specific real-time RT-PCR by Guthrie et al., 2013

    5. ANNEX V

      1. PART A

      2. PART B

    6. ANNEX VI