COUNTRY REPORTS ON AVIAN INFLUENZA FOR 004 BASED ON RESPONSES TO THE QUESTIONNAIRE Dennis J. Alexander and Ruth J. Manvell Community Reference Laboratory for Avian Influenza Veterinary Laboratories Agency Weybridge, New Haw, Addlestone, Surrey KT5 3NB, United Kingdom. INTRODUCTION Continuing the format adopted at the 7 th Meeting the information for this report was taken from answers supplied by National laboratories to the following questionnaire: *** AVIAN INFLUENZA. How many samples from which species of bird/type of poultry have been processed that would have resulted in the isolation of avian influenza viruses in and in? Example response: broilers 00 in 60 in turkeys 00 in 40 in 40 in s. State the number of influenza viruses isolated, their subtype, and the type of bird from which they were isolated. Example response: meat turkeys 3 x H6N x H9N waterfowl x H4N6, x H5N 3. For all influenza viruses isolated state type of poultry or species of bird and IVPI. For H5 and H7 isolates give amino acid sequence at the HA0 cleavage site and conclusion. Example response: Bird subtype IVPI HA0 cleavage site conclusion Turkeys H9N 0.00 nd LPAI feral duck H5N 0.00 PQRETR*GLF LPAI 4. Was any active surveillance for avian influenza carried out? If so give details of birds sampled, number of samples and results. *** 9
RESULTS A total of 33 questionnaires was sent to different laboratories in 30 countries. Responses were received for 9/5 EU countries [/7 laboratories]: Austria, Belgium, Cyprus, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece x, Ireland, Italy, Latvia, Netherlands, Poland, Slovakia, Slovenia, Sweden, UK Great Britain, UK Northern Ireland and from 3/5 [3/6 laboratories] non-eu countries: Croatia, Norway and Switzerland. The samples tested and the results for avian influenza are summarised in the following pages. VIRUS ISOLATION REPORTS BY COUNTRY AUSTRIA Samples tested BELGIUM Samples tested Type of bird Sample Method Number Poultry (chickens and turkey) Tissue 95 Cloacal swabs 0 Poultry (others) Tissue Psittacine Tissue 53 Duck and geese Cloacal swabs 4 pigeon Tissue 33 Pet birds Tissue 04 Tissue Cell culture 6 Quarantine birds Cloacal swabs 735 Wild birds Tissue 7 Influenza virus isolated Illegally imported eagle: x H5N Virus characterization Bird Subtype IVPI HA0 cleavage site conclusion eagle H5N.94 KRRKKR*GLF HPAI 0
CYPRUS Samples tested by inoculation into Chicken broilers tissue Chicken layers tissue 4 Falcons tissue 5 Flamingos tissue Hawks tissue 3 Partridges tissue 7 Pet birds, (parrots, canaries, finches) tissue 33 Pigeons tissue 7 Thrushes tissue 5 Various free living birds tissue 8. CZECH REPUBLIC Birds Sample Number Result chickens tissues 6 negative ducks tissues negative exotic birds faeces tissues 30 3 negative negative DENMARK chickens and hens tissue 660 psittacine and other caged birds tissue 3549 ducks and geese tissue 44 game birds tissue turkeys and ostriches tissue 60 pigeons tissue 30 wild birds faeces 3000* wild ducks 65 *in pools of 5
Influenza virus isolated Wild birds [All isolates were from wild ducks] x HN3, x H3N, x H3N?, x H5N (LPAI), x H6N, x H8N, x H8N4, 3 isolates with subtyping still pending - not H5/H7 (9.7% of the samples were positive by RT-PCR for influenza A) Characterisation of AIV isolates Bird subtype IVPI HA0 cleavage site conclusion Feral ducks H5N nd PQKETR*GLF LPAI ESTONIA broilers FINLAND Layers faecal samples 7 Turkeys faecal samples 8 Broilers 9 Water and shorebirds 309 Other wild birds 4 87 Partridges 8 Pigeons 4
FRANCE Exotics birds 5 tracheal swabs pooled tissues 5 Chickens/layers Ducks tracheal swabs pooled tissues tracheal swabs pooled tissues tracheal swabs pooled tissues 4 00 50 6 7 Pigeons 30 Swallow pooled tissues 4 Turkeys turkey breeders: 3 x H6N meat turkeys: x H6N Characterisation of AIV isolates tracheal swabs pooled tissues 70 75 3 Bird Subtype IVPI HA0 cleavage site Conclusion 3 isolates, and cleavage sites turkeys H6N 0.00 determined: PQVETR*GLF LPAI breeders PQIETR*GLF meat turkeys H6N 0.00 isolates with the same cleavage site : PQIETR*GLF LPAI GERMANY Samples tested Type of bird Sample Method Number broilers 00 60 turkeys 00 40 40 3
meat turkeys waterfowl 3 x H6N, x H9N x H4N6, x H5N Characterisation of AIV isolates Bird subtype IVPI HA0 cleavage site conclusion turkeys H9N 0.00 nd LPAI feral duck H5N 0.00 PQRETR*GLF LPAI GREECE - THESSALONIKI Broilers 95 40 Broiler breeders 33 Layers 87 Meat turkeys 8 7 IRELAND Broiler Breeders 0 tracheal swabs 0 Broilers 6 *Chickens 9 Pheasants 8 Ducks Pigeons 4 *Exotic Geese 3 Turkeys 6 Wild 8 Other 8 4
ITALY broilers pools of 7 40 turkeys pools of 4 pools of tracheal swabs 8 domestic duck pools of 4 pools of tracheal swabs 9 ostriches pools of 87 quail pools of 8 pools of faeces pigeons 7 parrots pools of faeces 6 geese pools of 3 pools of faeces Samples from wild birds: mallard (Anas platyrhynchos) 36 teal (Anas crecca) pintail (Anas acuta) gadwall (Anas strepera) shoveller (Anas clypeata) tufted duck (Aythya fuligula) 9 coot (Fulica atra) 8 dunlin (Calidris alpina) 7 black-headed gull (Larus ridibunot doneus) pools of tracheal swabs 0 cormorant (Phalacrocorax carbo) 3 Cetti s warbler (Cettia cetti) lapwing (Vanellus vanellus) 3 moorhen (Gallinula chloropus) From intensively reared birds turkeys x H7N3 quail x H7N3 From backyard flocks ducks x HN, x HN, x H4N, x H5N3, x H4N6, x H7N7, 3 x H0N4 geese x HN, x H3N8, x H4N, x H7N7, x H9N8, x HN, x HN9 From wild birds mallard (Anas platyrhynchos) x H4N6, 3 x H7N7, x H7N4 5
Characterisation of AIV isolates Bird subtype IVPI HA0 cleavage site conclusion turkey H7N3 0.00 PEIPKGR*GLF LPAI quail H7N3 Not done PEIPKGR*GLF LPAI duck HN Not done Not done - duck HN Not done Not done - duck H4N6 Not done Not done - duck H0N4 Not done Not done - duck H0N4 Not done Not done - duck H0N4 Not done Not done - duck H5N3 0.00 PQRDTR*GLF LPAI duck HN Not done Not done - duck H7N7 Not done Not done LPAI duck H4N Not done Not done - goose HN Not done Not done - goose H3N8 Not done Not done - goose H4N Not done Not done - goose HN9 Not done Not done - goose HN Not done Not done - goose H9N8 Not done Not done - goose H7N7 Not done PEIPKGR*GLF LPAI wild mallard H4N6 Not done Not done - wild mallard H7N7 Not done PEIPKGR*GLF LPAI wild mallard H7N7 Not done PEIPKGR*GLF LPAI wild mallard H7N7 Not done PEIPKGR*GLF LPAI wild mallard H7N4 Not done PEIPKGR*GLF LPAI LATVIA Broilers 40 Layers 45 Quail 6 6
NETHERLANDS chickens trachea swabs 40 lung/trachaea samples turkeys trachea swabs 40 ducks 0 cloaca swabs 0 faeces sample poultry lung/trachea samples 0 tracheal swabs 79 miscellaneous miscellaneous samples 57 unknown swabs 3 Exotic birds in quarantine faeces samples psittacines litter samples other birds Domestic duck x HN? Infection detected by RT- PCR faeces samples litter samples 30 06 607 Bird subtype IVPI HA0 cleavage site conclusion duck H5 ND* GLRNVPQKETR*GLF LPAI * No virus was isolated. pool was positive in M- and H5-RT-PCR. POLAND broilers pooled broiler breeders pooled turkeys pooled 3 ostriches pooled pigeons pooled black grouse pooled 4 sparrows faeces samples 6 storks 6 7
SLOVAKIA broilers tracheal swabs 4 layers 66 3 turkeys 34 ducks and geese 3 pigeons 6 wild birds 7 others faeces samples 4 96 3 SLOVENIA Surveillance in wild birds broilers 5 pigeons ducks grebes Cloacal swabs or samples from carcases tested by isolation in and RT- PCR Species of bird Number of samples waterfowl and pheasants ducks 39 geese 3 8
storks 3 herons swans grebes pheasants 5 other free-living birds* 30 Total number 4 *Sylvia attricapila, Sylvia borin, Panus major, Tardus merula, Erithacus rubecula, Acrocephalus schoenobaenus or detected SWEDEN broilers 6 imported layer breeders 8 breeders 3 layers 5 0 pigeons 8 wild birds 5 zoo birds 6 UK GREAT BRITAIN Samples tested Type of bird Sample Method Number tissues 66 70 chickens faeces swabs 5 3 turkeys pet birds tissues faeces swabs tissues 4 58 8 5 5 9
game birds pigeons waterfowl other faeces swabs tissues swabs tissues faeces swabs tissues swabs tissues cells 76 677 45 34 3 3 86 84 8 337 7 74 3 0 5 UK NORTHERN IRELAND chickens faeces tissues 30 3 4 pigeons tracheal swabs 3 45 turkeys 30 CROATIA broilers 39 broiler breeders layers 4 backyard chicken 3 pigeons 4 0
0 doves 7 ostriches 3 pheasant tissue sample swan tissue sample blackbird tissue sample vulture buzzard NORWAY Samples tested Bird Specimen Detection method Number Wild pigeon cloacal swab Inoc. in Wild duck Inoc. in 3 Wild duck RT-PCR 63 or detected SWITZERLAND. broilers 3 laying hens 6 duck duck DISCUSSION Responses The questionnaire for 004 reversed the trend of more countries responding each year seen over recent years. The responses for the last 4 years compared to the number of countries invited to complete the questionnaire have been: 000 9/9; 00 /9; 00 5/30; 003 8/30, but for 004 this was /30.
Samples tested The overall isolation attempts for avian influenza are summarised in Tables and for egg inoculations and Table 3 for inoculations. The overall total of 8,44 was considerably lower than totals of 45,866 in 004 and 35,374 for 00, but is more than twice the total of 8,498 in 00. Table Summary of virus isolation attempts in from tissue samples by countries responding to the questionnaire Bird Number countries Number chickens 0 544 turkeys 5 47 ducks & geese 7 63 game birds etc 8 87 ostriches 3 pigeons 4 340 cage, zoo & Q 0 4050 wild birds 0 TOTAL 6,655 Table Summary of virus isolation attempts in from, tracheal swabs and faecal samples by countries responding to the questionnaire Bird Number countries Number chickens 9 937 turkeys 8 666 ducks & geese 5 56 game birds etc 9 ostriches 87 pigeons 5 507 cage, zoo & Q 5 356 wild birds 9 4379 others 498 TOTAL,5
Table 3 Summary of virus isolation attempts from all samples* in cell cultures by countries responding to the questionnaire Type of bird Number countries reporting Number attempts chickens 95 turkeys 98 ducks & geese 5 game birds 37 pigeons cage, zoo, pet, quarantine etc 5 TOTAL 67 * tissues/tracheal swabs//faeces Viruses isolated Of the 4 laboratories responding 8 reported no isolations of AI viruses. HPAI The only isolations of HPAI in the EU in 004 were the H5N viruses obtained from the mountain hawk eagles smuggled from Thailand and detected at Brussels Airport in October 004 [see these proceedings]. LPAI H5 and H7 subtypes The H5 and H7 LPAI viruses isolated in the EU during 004 are summarised in Table 4. Table 4 Summary of H5 or H7 subtype LPAI viruses isolated. Subtype Country Bird Number HA0 cleavage site H5N Denmark PQKETR*GLF feral ducks Germany PQRETR*GLF H5N3 Italy domestic duck PQRDTR*GLF H5N? Netherlands domestic duck PQKETR*GLF H7N3 Italy meat turkeys PEIPKGR*GLF quail PEIPKGR*GLF H7N4 Italy feral mallard PEIPKGR*GLF H7N7 Italy domestic duck domestic goose feral mallards 3 PEIPKGR*GLF PEIPKGR*GLF PEIPKGR*GLF Other LPAI viruses A total of 38 LPAI influenza viruses of subtypes other than H5 or H7 was isolated from 5 countries (Table 3). Thirteen of these isolates were obtained from wild ducks and a further 5 from domestic ducks or geese. Isolates of H6N viruses were obtained from turkeys in France and Germany and two 3
H9N viruses from meat turkeys in Germany. There were no isolates of these viruses from chickens. Table 5 Summary of other LPAI viruses isolated by countries responding to the questionnaire Type of bird Subtype No. of isolates No. Countries turkeys H6N 8 H9N HN HN commercial ducks H4N H4N6 H0N4 HN? 3 commercial geese wild ducks HN H3N8 H4N H9N8 HN HN9 HN3 H3N H3N? H4N6 H6N H8N H8N4 untyped 3 4