{
  "url": "https://openlegislation.uk/eu-decision-2006-437/annex/chapter-V/division-3",
  "json_url": "https://openlegislation.uk/eu-decision-2006-437/annex/chapter-V/division-3.json",
  "legislation": "eudn/2006/437",
  "legislation_title": "Commission Decision of 4 August 2006 approving a Diagnostic Manual for avian influenza as provided for in Council Directive 2005/94/EC (notified under document number C(2006) 3477) (Text with EEA relevance) (2006/437/EC)",
  "legislation_url": "https://openlegislation.uk/eu-decision-2006-437",
  "path": "annex/chapter/V/division/3",
  "kind": "division",
  "number": "3",
  "heading": "Virus isolation in embryonated fowls' eggs",
  "text": "The clarified supernatant fluid must be inoculated in 0,1 to 0,2 ml amounts into the allantoic cavity of each of a minimum of four embryonated fowls' eggs that have been incubated for nine to 11 days. Ideally, these eggs must be obtained from a specific pathogen free (SPF) flock, but when this is impracticable eggs obtained from a flock shown to be free of antibodies to AI (serum antibody negative — SAN), may be used.\nThe inoculated eggs must be held at 37 oC and candled daily. Eggs with dead or dying embryos as they arise, and all remaining eggs six days after inoculation must be chilled to 4 oC and the allantoic- amniotic fluids tested for haemagglutination activity. If no haemagglutination is detected, this procedure must be repeated using undiluted allantoic/amniotic fluid as inoculum. When haemagglutination is detected the presence of bacteria must be excluded by culture. If bacteria are present, the fluids may be passed through a 450 nm membrane filter, further antibiotics added and inoculated into embryonated eggs as above.\nTo expedite diagnosis some laboratories have used two 3-day passages or 2-day and 4-day passages and reported comparable results to two 6-day passages, but this has not yet been fully evaluated.\nPositive fluids need to be tested for freedom from bacteria. If bacteria are present the fluids may be passed through a 450-nm membrane filter or centrifuged to remove bacteria and re-passaged in eggs after the addition of more antibiotics.",
  "extent": "E+W+S+N.I.",
  "status": null,
  "version": "current",
  "annotations": [],
  "outstanding_effects": [],
  "provenance": {
    "source": "legislation.gov.uk",
    "source_url": "https://www.legislation.gov.uk/eudn/2006/437/annex/chapter/V/division/3",
    "retrieved_at": "2026-11-01T15:17:33.742537+00:00"
  },
  "licence": "Open Government Licence v3.0",
  "licence_url": "https://www.nationalarchives.gov.uk/doc/open-government-licence/version/3/",
  "attribution": "Contains public sector information licensed under the Open Government Licence v3.0. Source: legislation.gov.uk, The National Archives."
}