Council Directive 89 /556/EEC of 25 September 1989 on animal health conditions applicable to intra-Community trade in and imports from third countries of embryos of the domestic animals of the bovine species has been incorporated into the national legislation by Royal Decree 855/1992 of 10 July laying down the animal health conditions applicable to intra-Community trade and to the imports from third countries of embryos of domestic animals of the species bovine.

Later, Council Directive 93 /52/EEC of 24 June amending Directive 89 /556/EEC, including in its field of application to embryos derived from certain techniques, with the addition of certain guarantees.

Changes are also made, setting the status of semen used for egg fertilisation and reflecting the new Community policy on foot-and-mouth disease.

On the other hand, Commission Decision 94 /113/EEC of 8 February sets out the additional guarantees for embryos subject to techniques requiring the penetration of the transparent zone and the fertilisation derivatives. Vitro ".

It is therefore necessary to introduce into Royal Decree 855/1992 of 10 July 1992 the amendments established by Directive 93 /52/EEC, which is transposed into national law by means of this provision, and by Decision 94 /113/EEC, in accordance with the State competence contained in Article 149.1.10. of the Constitution, in the field of foreign trade, and 149.1.16. of bases and coordination of health, and once the sector has been consulted.

By virtue of the proposal of the Minister for Agriculture, Fisheries and Food, in agreement with the Council of State and after deliberation by the Council of Ministers at its meeting on 6 May 1994,

D I S P O N G O:

Single item.

Royal Decree 855/1992 of 10 July 1992 laying down the animal health conditions applicable to intra-Community trade in and imports from third countries of animal embryos domestic of the bovine species, is amended as follows:

1. Article 1 (2) is worded as follows:

" 2. This Royal Decree shall not apply to embryos resulting from the transfer of nuclei. '

2. In Article 2, a paragraph (g) is added, with the following text:

"g) Embryo Production Equipment: the officially approved embryo collection team for in vitro fertilisation, in accordance with the conditions set out in the relevant Annex."

3. The first subparagraph of Article 3 (1) (a) is worded as follows:

" (a) must have been obtained as a result of artificial insemination or in vitro fertilisation, with semen from a donor of a semen collection centre approved by the competent authority for collection, treatment and storage of semen, or semen imported in accordance with the provisions of Royal Decree 877/1990 of 6 July 1990 laying down the animal health requirements applicable to intra-Community trade and imports of frozen meat of bovine animals. '

4. Article 4 is deleted.

5. In Article 5, a paragraph 4 is added, with the following text:

" 4. Authorisation shall only be granted to an embryo production team obtained by "in vitro" fertilisation, if the provisions of the corresponding Annex to this Royal Decree are complied with and if the embryo production team is able to comply with the corresponding provisions of this Royal Decree, and in particular paragraphs 1, 2 and 3 of this Article, which shall apply mutatis mutandis. '

6. Article 9 is structured in two paragraphs:

Paragraph 1 with the content of the current Article 9 and paragraph 2 with the following wording:

" 2. With regard to foot-and-mouth disease:

(a) From third countries which are vaccination against foot-and-mouth disease, only frozen embryos may be imported. Such embryos shall be stored under the conditions laid down for a minimum period of 30 days prior to dispatch.

(b) The donor animals shall come from a holding where no animal against foot-and-mouth disease has been vaccinated, during the 30 days prior to collection and which is not subject to any prohibition measure or quarantine. "

7. Article 11 is worded as follows:

" Article 11.

The principles and rules governing the organisation of veterinary checks for products from third countries introduced into the Community and the safeguard measures to be applied shall apply. apply. "

8. Articles 12 and 13 are deleted.

9. The Annexes are replaced by those of this Royal Decree.

Single end disposition.

This Royal Decree shall enter into force on the day following that of its publication in the "Official Gazette of the State".

Given in Madrid to May 6, 1994.

JOHN CARLOS R.

The Minister of Agriculture, Fisheries and Food,

LUIS MARIA ATIENZA SERNA

ANNEX A

CHAPTER I

Conditions for the authorization of embryo collection teams

In order to be authorised, each embryo collection team must meet the following requirements:

A) The collection, treatment and storage of embryos shall be carried out by a team veterinarian or, under his responsibility, by one or more competent technicians and trained in the hygiene methods and techniques.

B) Act under the general control of the official veterinarian and under his authority.

C) shall have at its disposal, for the examination, treatment and packaging of embryos, permanent or mobile laboratory facilities, composed of at least one working surface, a microscope and a team cryogenic.

D) You will have at your disposal, if the lab has a permanent site:

(a) A local for the handling of attached embryos, but physically separated from the area in which the donor animals are handled during collection.

(b) A local or area equipped for the cleaning and sterilisation of the equipment and material used for the collection and handling of embryos.

(c) If a micro-manipulation of the embryo which requires penetration of the transparent zone is to be performed, it must be carried out in facilities with adequate laminar flow. Such facilities shall be properly cleaned and disinfected between lots.

E) shall have at its disposal, in the case of a mobile laboratory, a part of the specially equipped vehicle, consisting of two separate sections:

a) A clean area, intended for the examination and manipulation of embryos, and

b) An area intended to house equipment and materials that come into contact with donor animals.

Every mobile laboratory will always be in contact with a permanent site laboratory, with a view to the sterilization of its equipment and to the supply of fluids and other products needed for collection and handling. of embryos.

In addition, in order for a team to be allowed to produce and treat embryos resulting from in vitro fertilisation or in vitro culture, the team must also meet the following additional requirements:

F) Personnel must be trained in appropriate laboratory and disease control techniques, especially in procedures for working under sterile conditions.

G) A transformation laboratory with a permanent location must be available to:

(a) With appropriate equipment and facilities, including a separate room for recovering oocytes from the ovaries, and separate rooms or areas for the transformation of oocytes and embryos, and for storage of embryos;

b) Dispose of laminar-flow facilities, under which all oocytes, semen and embryos must be treated; however, the centrifuge of semen may be carried out outside the laminar-flow facility provided it is take all necessary hygiene precautions.

H) When oocytes and other tissues are to be collected in a slaughterhouse, the equipment shall have at its disposal adequate equipment for the collection and transport of the ovaries and other tissues to the treatment laboratory in a hygienic manner. and secure.

CHAPTER II

Conditions relating to the collection, production, processing, storage and transport activities carried out by authorised embryo collection teams.

1. Collection and treatment.

A) The embryos shall be collected and processed by authorised collection teams, without coming into contact with other batches of embryos which do not comply with the requirements of this Royal Decree.

B) Embryos shall be collected in a place that is isolated from other parts of the enclosure or establishment and is in good conservation status and is easy to clean and disinfect.

(C) Embryos shall be treated (examined, washed, handled and placed in identified and sterile containers), either in a permanent laboratory or in a mobile laboratory located in an area not covered by the prohibition or quarantine.

(D) The material which is in contact with the embryos or the donor animal in the collection and treatment stages shall be disposable or subjected to adequate disinfection and sterilisation before reusing it.

E) Products of animal origin used during the collection of embryos and in the means of transport shall be from sources which do not pose a risk to the animal health status of the animals or receive prior treatment. to the use that eliminates that risk.

All means and solutions will be sterilized by applying authorized methods according to the recommendations of the annual International Embryo Transfer Society (IETS, International Embryo Transfer Society). According to the IETS manual, antibiotics may be added to the media.

F) The storage and transport containers will be properly disinfected or sterilized before each filling operation begins.

G) The cryogenic agent used has not previously been used for other products of animal origin.

H) Each embryo container, as well as the container in which embryos are stored and transported, shall be clearly marked by code so that the date of collection of the embryos can be easily determined, of the race and identification of the male and female donors and the registration number of the team. The form and characteristics of this code shall be determined in accordance with the procedure laid down by the Community authorities.

I) The embryo shall be washed at least ten times in special liquid, which shall be renewed each time and which, unless otherwise decided in application of paragraph (M), shall contain the trypsin, in accordance with the internationally recognised procedures. Each wash shall consist of a centesimal dissolution of the preceding washing, using a sterile micropipette to transfer the embryo each time.

J) After the last washing, it will be examined microscopically, with a minimum magnification of x 50, each of the embryos in all its surface, to check that the transparent zone is intact and there are no substances Attached to it.

Any micro-manipulation requiring the penetration of the transparent zone shall be carried out on the premises authorised for such purposes, and after the last washing and examination. Such micro-manipulation may not be performed in an embryo whose transparent area is not previously intact.

K) Each batch of embryos which has successfully passed the test set out in paragraph J) shall be deposited in a sterile container, which shall be marked in accordance with paragraph H) and sealed immediately thereafter.

L) If necessary, each embryo shall be frozen as soon as possible and stored in a local subject to the control of the team veterinarian and regularly inspected by the official veterinarian.

(M) In accordance with the provisions laid down by the Community authorities, the competent bodies of the Autonomous Communities shall draw up a protocol on rinsing and washing liquids, washing techniques, and, if necessary, enzyme treatments, as well as the means of conservation authorised for transport.

Until the adoption of a protocol on enzyme trations, national rules on the use of trypsin shall continue to apply.

N) Each collection team shall take routine samples of the products resulting from the official examination of the equipment, with rinsing and washing liquids, disintegrated embryos, infected eggs, etc., for the detection of possible bacterial and viral contamination. The procedure for the collection of samples, the criteria for the examinations and the levels to be achieved shall be those laid down by Community legislation, in accordance with the intended Community procedure.

O) Each collection team must keep track of the embryo collection activities carried out in the last 12 months before and after storage. Such registration shall include:

a) The race, age and identification of the corresponding donor animals.

b) The place of collection, treatment and storage of the embryos collected by the team.

c) The identification of the embryos with details of their destination, if known.

(d) Details of micro-manipulation techniques that require penetration of the transparent zone or other techniques, such as in vitro fertilisation or in vitro culture, to which the embryos have been subjected. In the case of embryos resulting from 'in vitro' fertilisation, identification shall be provided from the batch, but the date and place of collection of the ovaries or oocytes should be mentioned. It should also be possible to identify the herd of origin of the donor animals.

The conditions set out in paragraphs (A) to (O) shall apply, where appropriate, to the collection, treatment, storage and transport of ovaries, oocytes and other tissues for use for in vitro fertilisation or in vitro culture. The following additional conditions shall also apply:

P) When the ovaries and other tissues are to be collected in a slaughterhouse, the slaughterhouse must be officially approved and under the control of an official veterinarian responsible for the ante-mortem and post-mortem inspections of the donors.

Q) Materials and equipment that come into direct contact with the ovaries and other tissues must be sterilized prior to their use and, after this process, should be used only for these purposes. Different equipment shall be used to treat oocytes and embryos from different batches of donor animals.

R) The entry into the treatment laboratory of the ovaries and other tissues shall not be permitted until the "post-mortem" inspection of the lot has been completed. If relevant signs of disease are found in the donor lot or in any animal slaughtered in the same slaughterhouse on the same day, all tissues in that lot must be found and removed.

S) The washing and examination procedures set out in paragraphs (I) and (J) shall be carried out after the cultivation procedure has been completed.

T) Any micro-manipulation requiring penetration of the transparent zone shall be carried out in accordance with the provisions of paragraph (J), once the procedures laid down in paragraph S have been completed.

U) Only embryos from the same batch of donors shall be stored in the same ampoule or straw.

2. Storage.

The embryo collection or production teams shall ensure that the embryos are stored at appropriate temperatures in premises approved for that purpose by the Autonomous Community.

To be authorized, those locales must:

(a) Contar at least one local that can be locked and used exclusively for the storage of embryos.

b) Be easy to clean and disinfect.

(c) Dispose of records in which all entries and exits of embryos are to be permanently entered. These records shall specify in particular the final destination of the embryos.

d) Be inspected by the official veterinarian.

The competent authority of the Autonomous Communities may authorise the storage of semen complying with the provisions of Royal Decree 877/1990.

3. Transport.

Embryos intended for trade shall be transported in satisfactory hygienic conditions, in sealed containers from the authorised storage premises until their arrival at the place of destination.

The containers will be marked in such a way as to match the number listed in the health certificate.

ANNEX B

Conditions for donor animals to be required

1. For the purpose of embryo collection, donor animals shall meet the following requirements:

(a) For the preceding six months, they shall have remained in the territory of the Community or of the third country of collection;

(b) They shall be present in the herd of origin for a minimum of 30 days prior to collection;

c) herd procedures:

-Officially tuberculosis-free.

-Officially free of brucellosis or brucellosis-free.

-Indemnes of enzootic bovine leukosis.

By virtue of an exception provided for in the third indent, donor animals may come from a herd (or flocks) which is not free but for which it is certified that there has been no clinical case of leukosis. enzootic bovine in the last three years;

(d) For the previous year, no infectious bovine rhinotracheitis or infectious purulent vulvovagitis clinical signs have been present in a herd (or herds) in the herd.

2. On the day of embryo collection, the donor female:

(a) Be found in an establishment which is not subject to prohibition or veterinary quarantine measures;

b) You must not show any clinical signs of disease.

3. The above conditions shall also apply to live animals intended to be converted into oocyte donors by means of egg collection or ovariotomy.

4. In the case of donors of ovaries and other tissues to be collected after slaughter in a slaughterhouse, those animals shall not have been selected for slaughter within a national programme for the eradication of a disease or come from a holding subject to restrictions due to the presence of an epizootic disease.

5. The slaughterhouse in which the ovaries and other tissues are collected shall not be located in an area subject to prohibition or quarantine measures.

ANNEX C