Highly Pathogenic Avian Influenza (H7N7) in Italy

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1 Highly Pathogenic Avian Influenza (H7N7) in Italy STANDING COMMITEE ON PLANTS, ANIMALS, FOOD AND FEED Section Animal Health and Welfare st June 26

2 H7N7 HPAI Outbreak in Emilia Romagna - 6/ Organic free-range laying hens holding in Ferrara province (Emilia Romagna region) 7. laying hens 29 April 26 - Farmer reported increased daily mortality 3 April 26 - The National Reference Laboratory confirmed the positivity for Influenza A virus subtype H7

3 H7N7 HPAI Outbreak in Emilia Romagna - 6/ May 26 Beginning of depopulation and carcasses disposal 2 May 26 The National Reference Laboratory identifies the H7N7 HPAI virus 2 May 26 End of depopulation and disposal of carcasses (rendering)

4 Office and warehouse Run

5

6 Daily Mortality

7 Weekly Egg Production

8 Consumption of feed Data Weekly Consumption (kg) Average daily consumption (kg) 29/2/26-6/3/26 39,72 9,98 7/3/26-3/3/26 39,5 9,93 4/3/26-2/3/26 39,3 9,9 2/3/26-27/3/26 38,88 9,84 28/3/26-3/4/26 38,39 9,77 4/4/26 - /4/26 37,69 9,67 /4/26-7/4/26 37,2 9,6 8/4/26-24/4/26 37,3 9,59

9 Protection and Surveillance Zones

10 Result of epidemiological enquiry Basing on epidemiological data and phylogenetic analysis: Possible virus introduction as Low Pathogenicity through direct contact between wild birds and free-range laying hens Likely mutation from Low to High pathogenicity strain, with increased mortality and decrease in egg production Sequencing of the virus showed two different pathotypes (HPAI and LPAI)

11 Epidemiological contacts Feed lorry visited the infected holding on 4 April The vehicle came from a feed factory annexed to a laying hens holding, in Treviso province (Veneto region) Table eggs produced before the suspect confirmation were sent to a packaging centre in Cremona province (Lombardia region) The vehicle stopped for egg load in another holding in Bologna province (Emilia Romagna region) before reaching the packaging centre Lorry of the rendering plant visited the infected holding on 9 April Vehicle collected the carcasses in other 3 poultry farms in Padova province (Veneto region), after visiting the infected farm

12 feed factory with poultry farm egg packaging centre poultry farms outbreak laying hens holding

13 Control measures implemented in contact holdings All the contact holdings were put under official restrictions and official controls on reared birds were performed as provided for by the national provisions 3 blood samples/shed 3 tracheal swabs/shed 5 dead birds/shed After 2 days from the last at-risk contact, and after the negative results of the official tests the restrictive measures were lifted in all the contact holdings

14 Control measures at a national level In all layer holdings located in Densely Populated Poultry Areas (DPPAs) 3 tracheal swabs and 3 blood samples for shed Clinical inspection and submission of at least 5 dead birds for each affected shed in case of increased mortality, drop in egg production and/or feed consumption

15 Control measures at a national level Pre-moving testing of pullets and laying hens in Densely Populated Poultry Areas (DPPAs) Pre-moving testing of fattening turkeys before loading for slaughter in DPPAs

16 Control measures at a national level (DPPA) No. Of PCR Analysis No. Of Confirmed PCR Species No. Of Farms No. Of Animals M-gene PCR H5 H7 H5 H7 FATTENING TURKEY LAYING HENS TOTAL Layers holdings Fattening turkeys holdings

17 Control measures at a national level Enhanced biosecurity measures Functional separation of production between Emilia Romagna and the other regions

18 Phylogenetic analysis The HA gene of the virus A/chicken/Italy/6VIR-873/26 (H7N7) clusters with H7 subtypes detected in Africa and Asia, between 2 and 23 in both wild and domestic birds The NA gene of the virus A/chicken/Italy/6VIR-873/26 (H7N7) clusters with AIVs belonging to HN7 and H7N7 subtypes detected in Europe, between 2 and 25 in wild and domestic birds and seals The HA gene resulted to be low related to other HPAIVs recently identified in Italy and Europe and to the H7N7 LPAI virus responsible for the Ferrara outbreak of 5 April 26 (similarity lower than 93%) No additional glycosylation sites or molecular markers of adaptation to mammalian hosts have been identified in the HA gene.

19 . A/Eurasian_teal/Italy/439-7/25_H7N7 A/mallard/Italy/63-5/27_H7N A/Mallard/Italy/ /26_H7N7 A/mallard/Italy/63-2/27_H7N3 A/duck/Bangladesh/4VIR2-/23_H7N5 A/duck/Mongolia/9/28_H7N9 A/velvet_scoter/Mongolia/986V/29_H7N3 A/duck/Mongolia/29/2_H7N9 A/Netherlands/29/3_H7N7 A/mallard/Askania-Nova/39-5-2/2_H7N7 A/mallard/Novomyhalivka/-23-2/2_H7N7 A/Mallard/Italy/ /26_H7N7 A/ostrich/South_Africa/ORD/22_H7N A/Mallard/Sweden/5/2_H7N7 A/mallard/Italy/43/_H7N3 A/chicken/Pakistan/CR2/95_H7N3 A/chicken/Rizhao/875/23_H7N9 A/shoveler/Italy/64-3/27_H7N3 A/duck/Italy/6VIR-64-2/26 A/teal/Italy/6VIR345-29/26/H7 A/mallard/italy/ /28_H7N A/chicken/Pakistan/34669/995H7N3 A/shoveler/Egypt/7-NAMRU3/27_H7N3 A/garganey/Italy/699-2/29 A/Netherlands/3/3_H7N7 A/Mallard/Italy/497-29/26_H7N7 A/mallard/Italy/73/9_H7N A/chicken/Italy/ /27 A/chicken/Italy/ /27 A/ostrich/South_Africa/MKT/22_H7N A/chicken/Murree/NARC-/995_H7N3 A/mallard/Italy/794-8/28_H7N A/shoveler/Italy/2698-3/26_H7N7 A/shoveler/Egypt/25-NAMRU3/27_H7N9 A/chicken/England/283/25_H7N7 A/northern_shoveler/Egypt/EMC-/2_H7 A/shoveler/Egypt/4879-NAMRU3/26_H7N A/Chicken/NL-Barneveld/ /25_H7N7 A/turkey/Germany/R/2_H7N7 A/Mallard/Italy/2699-6/26_H7N7 A/duck/Italy/4VIR-383/24_H7N7 A/Mallard/Italy/ /26_H7N7 A/Shoveler/Italy/2698-3/26_H7N7 A/silkie_chicken/Wenzhou/86/23_H7N7 A/teal/Crimea/227/28_H7N8 A/duck/Turkey/55/Cetinkaya/49/26_H7N A/environment/Bangladesh/97/22_H7N9 A/duck/Italy/6VIR-64-/26 A/mallard/Poland/6/9 A/mallard/Italy/336/7_H7N3 A/mallard/Republic_of_Georgia/2/2_H7N7 A/turkey/Germany/R/_H7N7 A/ruddy_shelduck/Mongolia/9/2_H7N7 A/ruddy_shelduck/Mongolia/29/2_H7N A/mallard/Italy/VIR-54/_H7N7 A/Shoveler/Italy/377-6/26_H7N7 A/ostrich/South_Africa/KVL/22_H7N A/mallard/Italy/738-27/29 A/turkey/Germany-NI/R534/23_H7N7 A/Anas_crecca/Spain/46/28_H7N9 A/chicken/Pakistan/447/95_H7N3 A/chicken/Rizhao/87/23_H7N9 A/chicken/Italy/6VIR-873/26_H7N7 A/chicken/England/58-46/28_H7N7 A/garganey/Crimea/227/28_H7N8 A/mallard/Iran/V3/4_H7 A/mallard/Nizhnegorsk/3-6-/2_H7N7 A/teal/Italy/VIR-792/_H7N6 A/mallard/Sweden/5/22_H7N7 H7N HPAI-LPAI Italy A/Mallard/Italy/44-8/25_H7N7 A/Mallard/Italy/497-35/26_H7N7 A/teal/Italy/794-3/28_H7N A/mallard/Sweden/333/23_H7N7 A/Netherlands/27/3_H7N7 A/mallard/Republic_of_Georgia/3/2_H7N3 A/ostrich/South_Africa/FWS/22_H7N A/ruddy_shelduck/Mongolia/89/2_H7N A/mallard/Sweden/24987/2_H7N7 A/common_goldeneye/Mongolia/27/2_H7N A/Mallard/Italy/2699-5/26_H7N7 A/mallard/Netherlands/2/2_H7N3 A/mallard/Italy/ /28_H7N A/mallard/Italy/99/_H7N3 A/pigeon/Wenzhou/559/23_H7N7 A/northern_shoveler/Egypt/EMC-/22_H7N A/mallard/Poland//8_H7N7 A/Shoveler/Italy/ /26_H7N7 A/chicken/Karachi/NARC-/24_H7N3 A/ruddy_shelduck/Mongolia/436/2_H7N H7N7 HPAI Italy 23 A/chicken/England/26352/25_H7N7 H7N3 Italy 27-2 A/chicken/NL-Barneveld/ /25_H7N7 A/Fujian//23_H7N9 A/sacred_ibis/South_Africa/27/22_H7N A/Eurasian_teal/Italy/533/25_H7N7 A/guinea_fowl/Italy/47/28_H7N A/shoveler/Egypt/597-NAMRU3/24_H7N A/shoveler/Italy/6323-/27_H7N3 H7N3 Italy A/mallard/Italy/33/_H7N3 A/shellduck/South_Africa/DLH/22_H7N8 A/mallard/Italy/64-4/27_H7N3 A/mallard/Sweden/227/23_H7N7 A/mallard/Korea/822/_H7N7,98,79,23,89,92,99,79,83,8,9,94,88,9,99,99,96,97,9,9,99,75,75,98,7,94,5,89,75,97,76,95,84,9,92,88,99,93,9,73,92,66,84,9,7,76,34,87,92,89,76,96,78,99,9,98,82,96,79,75,8,77,88,76,97,98,99 A/mallard/Italy/63-5/27_H7N A/Mallard/Italy/ /26_H7N7 A/mallard/Italy/63-2/27_H7N3 A/duck/Bangladesh/4VIR2-/23_H7N5 A/velvet_scoter/Mongolia/986V/29_H7N3 A/duck/Mongolia/29/2_H7N9 A/mallard/Askania-Nova/39-5-2/2_H7N7 A/mallard/Novomyhalivka/-23-2/2_H7N7 A/Mallard/Italy/ /26_H7N7 A/ostrich/South_Africa/ORD/22_H7N A/shoveler/Italy/64-3/27_H7N3 A/duck/Italy/6VIR-64-2/26 A/teal/Italy/6VIR345-29/26/H7 A/mallard/italy/ /28_H7N A/shoveler/Egypt/7-NAMRU3/27_H7N3 A/garganey/Italy/699-2/29 A/Mallard/Italy/497-29/26_H7N7 A/ostrich/South_Africa/MKT/22_H7N A/shoveler/Italy/2698-3/26_H7N7 A/chicken/England/283/25_H7N7 A/northern_shoveler/Egypt/EMC-/2_H7 A/shoveler/Egypt/4879-NAMRU3/26_H7N A/Chicken/NL-Barneveld/ /25_H7N7 A/Mallard/Italy/2699-6/26_H7N7 A/duck/Italy/4VIR-383/24_H7N7 A/Mallard/Italy/ /26_H7N7 A/Shoveler/Italy/2698-3/26_H7N7 A/duck/Turkey/55/Cetinkaya/49/26_H7N A/environment/Bangladesh/97/22_H7N9 A/duck/Italy/6VIR-64-/26 A/mallard/Poland/6/9 A/mallard/Republic_of_Georgia/2/2_H7N7 A/ruddy_shelduck/Mongolia/9/2_H7N7 A/ruddy_shelduck/Mongolia/29/2_H7N A/mallard/Italy/VIR-54/_H7N7 A/Shoveler/Italy/377-6/26_H7N7 A/ostrich/South_Africa/KVL/22_H7N A/mallard/Italy/738-27/29 A/turkey/Germany-NI/R534/23_H7N7 A/chicken/Italy/6VIR-873/26_H7N7 A/mallard/Nizhnegorsk/3-6-/2_H7N7 A/teal/Italy/VIR-792/_H7N6 A/Mallard/Italy/497-35/26_H7N7 A/mallard/Republic_of_Georgia/3/2_H7N3 A/ostrich/South_Africa/FWS/22_H7N A/ruddy_shelduck/Mongolia/89/2_H7N A/common_goldeneye/Mongolia/27/2_H7N A/Mallard/Italy/2699-5/26_H7N7 A/mallard/Italy/ /28_H7N A/northern_shoveler/Egypt/EMC-/22_H7N A/mallard/Poland//8_H7N7 A/ruddy_shelduck/Mongolia/436/2_H7N H7N7 HPAI Italy 23 A/chicken/England/26352/25_H7N7 A/chicken/NL-Barneveld/ /25_H7N7 A/sacred_ibis/South_Africa/27/22_H7N A/shoveler/Italy/6323-/27_H7N3 A/shellduck/South_Africa/DLH/22_H7N8 A/mallard/Italy/64-4/27_H7N3 A/mallard/Korea/822/_H7N7,98,92,99,8,9,94,9,96,99,75,98,7,94,75,97,76,95,84,92,88,93,9,73,9,34,87,89,96,78,98,79,88,76,98,99 ML phylogenetic tree of the HA gene

20 H7N7 HPAI Outbreak in Emilia Romagna -6/2 Meat turkey holding in Ferrara province (Emilia Romagna region) The farm belongs to a vertical integrated group male turkeys (59 days old) Within the Protection Zone of the first H7N7 HPAI outbreak (6/)

21 Protection and Surveillance Zones

22 H7N7 HPAI Outbreak in Emilia Romagna - 6/2 3 May 26 Reported increased mortality in a single shed (n.2) 6 May 26 National Reference Laboratory confirmed the positivity for H7N7 HPAI virus 8 May 26 End of depopulation and carcass disposal

23 Daily mortality (shed n.2) Increased mortality in shed n.2 leads to suspect AI introduction

24

25

26 Result of epidemiological enquiry So far the introduction path of the virus remains unknown Pre-emptive killing in a single contact holding (same company; proximity) 7 May 26 Ministry of Health orders the pre-emptive slaughter of the turkey holding 8 May 26 Depopulation begins in the contact holding

27 Phylogenetical analysis The HA gene of the virus A/turkey/Italy/6VIR-225/26 (H7N7) shows a similarity of 99.6% with the H7N7 HPAI virus isolated in the previous outbreak (6/) clustering with H7 subtypes collected in Africa and Asia, between 2 and 23 in both wild and domestic birds; No additional glycosilation sites or molecular markers of adaptation to mammalian hosts have been identified in the HA gene.

28 ML phylogenetic tree of the HA gene

29 Thank you

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