92/532/EEC: Commission Decision of 19 November 1992 laying down the sampling plans and diagnostic methods for the detection and confirmation of certain fish diseases

• Published date: 21 November 1992
• Subject Matter: Veterinary legislation
• Official Gazette Publication: Official Journal of the European Communities, L 337, 21 November 1992

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31992D0532

92/532/EEC: Commission Decision of 19 November 1992 laying down the sampling plans and diagnostic methods for the detection and confirmation of certain fish diseases

Official Journal L 337 , 21/11/1992 P. 0018 - 0027
Finnish special edition: Chapter 3 Volume 46 P. 0019
Swedish special edition: Chapter 3 Volume 46 P. 0019

COMMISSION DECISION of 19 November 1992 laying down the sampling plans and diagnostic methods for the detection and confirmation of certain fish diseases (92/532/EEC)

THE COMMISSION OF THE EUROPEAN COMMUNITIES,

Having regard to the Treaty establishing the European Economic Community,

Having regard to Council Directive 91/67/EEC of 28 January 1991 concerning the animal health conditions governing the placing on the market of aquaculture animals and products (1), and in particular Article 15 thereof,

Whereas, in accordance with Article 15 of Directive 91/67/EEC sampling plans and diagnostic methods to be applied for the detection and confirmation of diseases in aquaculture animals shall be established;

Whereas the Scientific Veterinary Committee, established by Commission Decision 81/651/EEC (2) has been consulted;

Whereas the measures provided for in this Decision are in accordance with the opinion of the Standing Veterinary Committee,

HAS ADOPTED THIS DECISION:

Article 1

The sampling plans and diagnostic methods for the detection and confirmation of infectious Haematopoietic Necrosis (IHN) and Viral haemorrhagic Septicaemia (VHS) are laid down in the Annex.

Article 2

This Decision is addressed to the Member States. Done at Brussels, 19 November 1992. For the Commission

Ray MAC SHARRY

Member of the Commission

(1) OJ No L 46, 19. 2. 1991, p. 1. (2) OJ No L 233, 19. 8. 1981, p. 32.

ANNEX

PART I

SAMPLING AND TESTING PROCEDURES FOR VHS AND IHN MONITORING I. Sampling 1. Sampling time

Farms are inspected clinically at least twice per year during the period October to June or whenever the water temperature is below 14 °C. Intervals between inspections must be at least four months. All production units (ponds, tanks, aquaria, netcages, etc.) are inspected for the presence of dead, weak or abnormally behaving fish. Particular attention has to be paid to the water outlet area (if feasible) where weak fish tend to accumulate because of the water current.

2. Selection and collection of samples

Thirty to 150 fish and/or ovarian fluid samples are collected for examination in connection with the inspections according to Table 1. If rainbow trout are present fish of that species should make up the whole sample. If rainbow trout are not present the sample has to contain fish of all other species present whenever these species are susceptible to VHS and/or IHN as listed in Annex A of Council Directive 91/67/EEC concerning the animal health conditions governing the placing on the market of aquaculture animals and products. The species have to be equally represented in the sample. During the initial four-year control period which precedes achievement of approved status the sample size is 150 in order to ensure detection at a 95 % confidence level of virus carriers at a carrier prevalence of 2 %. During the subsequent years (maintenance of approved status) the sample size can be reduced to 30 to ensure detection at a 95 % confidence level of virus at a prevalence of 10 %.

In farms which have a documented history of freedom from VHS and IHN (based on a regular official health inspection programme) the small sample size can be used also during the initial four-year control.

If more than one water source is utilized for fish production, fish representing all water sources may be included in the 150 or 30 fish-sample. If weak, abnormally behaving or freshly dead (not decomposed) fish are present, these must primarily be included in the sample. If such fish are not present the sample must be composed of normally appearing, healthy fish collected in such a way that all parts of the farm as well as all year classes are represented in the sample.

3. Preparation and shipment of samples from fish

Before shipment or transfer to the laboratory pieces of the organs to be examined are removed from the fish with sterile scissors and forceps and transferred to plastic tubes containing transportation medium i.e. cell culture medium with 10 % calf serum and antibiotics. The combination of 200 iu penicillin, 200 mg streptomycin, and 200 mg kanamycin per millilitre (ml) can be recommended but other antibiotics of proven efficiency may be used as well. The tissue material to be examined is spleen, anterior kidney, encephalon and, in some cases, ovarian fluid (Table 1).

Organ pieces from 5 to 10 fish (Table 1) may be collected in one tube and represent one pooled sample. The tissue in each sample should weigh a minimum of 1 gram (g) and such that the final dilution 1: 10.

The tubes are placed in insulated containers (for instance thick-walled polystyrene boxes) together with sufficient ice of 'frost blocks' to ensure chilling of the samples to between 0 and 5 °C during transportation to the laboratory. Freezing must be avoided.

The virological examination must be started as soon as possible and not later than 48 hours after the collection of the samples. If the fish to be examined are less that 6 centimetres (cm) in length, the whole fish may be shipped to the laboratory in plastic bags chilled as mentioned above.

4. Collection of supplementary diagnostic material

According to agreement with the involved diagnostic laboratory, other fish tissues may be collected as well and prepared for supplementary examinations.

II. Preparation of samples for virological examination

1. Homogenization of organs

In the laboratory the tissue in the tubes must be completely homogenized (either by stomacher, blender or mortar and pestle) and subsequently suspended in the original transport medium. If a sample consisted of whole fish, i.e. fish less than 6 cm long, these are minced with sterile scissors after removal of the body behind the gut opening, homogenized as described above and suspended at a 1: 10 ratio in transport medium.

2. Centrifugation of homogenate

The homogenate is centrifuged in a refrigerated centrifuge at 2 - 5 °C at 2 000 to 4 000 x g for 15 minutes and the supernatant collected for examination.

If shipment of the sample has been made in a transport medium (i.e. with exposure to antibiotics) the supernatant...