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AVIAN INFLUENZA (157): ITALY, LPAI H5N2, H7N3

Gert van der Hoek

In Memoriam - Editor, Senior Moderator
Archive Number 20070919.3113
Published Date 19-SEP-2007
Subject PRO/AH/EDR> Avian influenza (157): Italy, LPAI H5N2, H7N3



AVIAN INFLUENZA (157): ITALY, LPAI H5N2, H7N3
*********************************************
A ProMED-mail post
<http://www.promedmail.org>
ProMED-mail is a program of the
International Society for Infectious Diseases
<http://www.isid.org>

Date: Tue 18 Sep 2007
From: Giuseppe Michieli <giuseppe.michie@libero.it>


I would like to submit to you recent reports posted by IZSVE (Istituto
Zooprofilattico delle Venezie-Legnaro-Padova, Italy, OIE (Office
International des Epizooties; World Organization for Animal Health)
Reference Laboratory for avian influenza) regarding multi-focal epizootics
of LPAI [low pathogenic avian influenza] A/H5N2 and A/H7N3 in several
northern provinces of Italy, which have appeared during the last few weeks.
Animals affected were mainly birds in farms (turkeys, broilers, ducks,
geese) as well as free-range poultry in vocational farms.

The reports are dispatched in PDF format and contain analytical tables with
phylogenetic trees of A/H5 and A/H7 viral strains isolated at infected
farms (there were only asymptomatic animals, or -- in one instance -- mild,
subclinical signs). The documents include maps and description of outbreaks
with the number of susceptible animals:
http://www.izsvenezie.it/dnn/Default.aspx?tabid=312 (Epidemiological
situation on 19 Sep 2007, in English and Italian).

I am a bit surprised because there isn't -- to date -- any official report
to OIE. Epizootics recur for several months but no OIE confirmation reports
were published on the WAHID (World Animal Health Information Database) site.

Northern provinces of Italy contain larger poultry industrial farms with
extremely high numbers of birds.

--
Giuseppe Michieli (not a health care worker)
Padova (Padua)
Italy
http://hygimia69.blogspot.com/
giuseppe.michie@libero.it

[The following documents can be downloaded from the useful English website
above:

1. Low pathogenic avian influenza in Italy: Lombardia, Veneto, Emilia
Romagna, Basilicata Region (situation on 17 Sep 2007).
2. LPAI in Italy: maps of outbreaks (Italy; Lombardia Region; Veneto
Region; Emilia Romagna Region; Basilicata Region).
3. Low pathogenic avian influenza in Italy: the epidemiological situation,
(22 pages, updated 10 Sep 2007).

Italy's most recent report to the OIE on avian influenza dates back to 21
Sep 2006; see <http://www.oie.int/eng/info/hebdo/AIS_77.HTM#Sec11.

http://www.promedmail.org/pls/askus..._BACK_PAGE,F2400_P1001_PUB_MAIL_ID:1000,39379
 
Re: AVIAN INFLUENZA (157): ITALY, LPAI H5N2, H7N3

LPAI H7N3 in Italy: the epidemiological situation​
Since the end of May, 2007, 13 low pathogenicity (LP) avian influenza (AI)
outbreaks caused by a type A AI virus of the H5 - H7 subtype have occurred in Italy
(Table 1). Eight LPAI outbreaks were identified in back-yard flocks or in dealer
farms supplying the rural and hobby poultry sector (last outbreak identified on 7
September 2007 in Mantova Province). On 22 August 2007, an H7N3 LPAI virus
was isolated in a meat turkey holding located in the Municipality of Paderno
Franciacorta in the Province of Brescia, Lombardia Region. From 22 to 7 September
2007, another 3 LPAI infected meat turkey holdings were identified in the same
Region.
Phylogenetic analysis performed on the HA gene indicated that the HA gene of the
virus isolated in meat turkeys in Lombardy Region was part of the Eurasian lineage
of H7 viruses and it was identical to the virus isolated in Rogno outbreak (100%
homology). Both viruses were closely related (HA homology 99.8%) to the H7N3
virus identified on 19 July 2007 in the outbreak of Fiesso Umbertiano, Rovigo
province in the Veneto Region. Based on the HA and NA sequences, these two
isolates did not appear closely related to the previous H7N3 LPAI virus, isolated
during an outbreak in rural chickens last May in Venice province, Veneto Region.
For HA and NA homology was 94.5% and 93.5%, respectively. Similarly, all these
H7N3 LPAI viruses isolated in 2007 were unrelated to the H7N3 virus which caused
the 2002-2004 LPAI epidemic in Italy.
Furthermore, on 14 August 2007, an LPAI virus of the H5N2 subtype was isolated in
a free-range goose and duck breeder flock in Emilia-Romagna Region. The HA gene
of this virus strain was not phylogenetically related (93.2% homology) to the H5N2
LPAI virus that caused the 2005 epidemic in Lombardia Region, whereas the HA
gene showed a higher level of homology (98.5%) with H5N2 LPAI viruses isolated
from wild waterfowl in Germany in 2005.
The geographical distribution of LPAI outbreaks in Italy is illustrated in figure 1.​
Pagina 3 di 22​
Table 1- LPAI outbreaks in Italy: the epidemiological situation.​
Region Province Municipality Date of
confirmation
Type of
production
N of
Birds
LPAI
virus
subtype​
Veneto VE Santa Maria di
Sala
24/05/2007 Rural flock
(ducks, broilers,
layers, etc.)
960 H7N3
Veneto RO Fiesso
Umbertiano
19/07/2007 Dealer farm
(ornamental
birds)
2.139 H7N3
Basilicata PZ Ruoti 31/07/2007 Rural flock
(chikens, turkeys,
geese)
75 H7
Emilia
Romagna
RA Lugo di Romagna 14/08/2007 Breeder flock
(geese and ducks)
- free range
10.150 H5N2
Emilia
Romagna
RA Conselice 17/08/2007 Dealer farm
ducks, broilers,
layers)
215 H7
Emilia
Romagna
RA Lugo di Romagna 21/08/2007 Dealer farm
(guinea fowl,
broilers)
7.100 H7
Lombardia BS Paderno
Franciacorta
22/08/2007 Meat turkey farm 11.400 H7N3
Lombardia BS Paderno
Franciacorta
22/08/2007 Meat turkey farm 15.860 H7
Lombardia BG Filago 24/08/2007 Hobby flock
(ornamental
birds)
1.446 H7
Lombardia CR Volongo 27/08/2007 Meat turkey farm 7.898 H7
Lombardia BS Coccaglio 31/08/2007 Meat turkey farm 11.697 H7
Lombardia BG Rogno 03/09/2007 Dealer farm 4.157 H7N3
Lombardia MN Castel Goffredo 07/09/2007 Dealer farm 25.300 H7​
Pagina 4 di 22​
Figure 1. Location of LPAI outbreaks in Italy​
Pagina 5 di 22​
VENETO REGION OUTBREAKS​
Since 24 May, 2007, two LPAI outbreaks have been confirmed in the Veneto Region
(Fig. 2). Outbreak data are summarised in Table 2.​
Table 2 ? Confirmed outbreaks of LPAI H7N3 in the Veneto Region.​
Date of confirmation Subtype Diagnosis Municipality No. of birds
24 May, 2007 H7N3 LPAI
Serological
Virological
Santa Maria di Sala
(VE)
960
19 July, 2007 H7N3 LPAI
Serological
Virological
Fiesso Umbertiano
(RO)
2.139​
Figure 2. Location of LPAI outbreaks in the Veneto Region.​
Pagina 6 di 22​
The SANTA MARIA DI SALA (VE) outbreak​
On 24 May 2007, during national avian influenza monitoring plan activities,a
number of birds on a rural farm in the municipality of Santa Maria di Sala (VE) (Fig.
3) tested serologically and virologically positive for H7N3 LPAI (cleavage site
typical of the LPAI strains: PEIPKGRGLF).
At the moment of sampling on 21 May, 2005, all the birds on the farm appeared
clinically healthy. Epidemiological investigation did not reveal the sale of birds
during the ?high risk? period. The farm in question works in the farm tourism sector
and most of the poultry raised is consumed internally. There were 960 birds on the
farm at the moment of the outbreak: 97 ducks, 46 geese, 670 broilers, 50 laying hens
and 97 guinea fowls. The Veneto Region issued eradication measures as specified by
EU Directive 94/2005/CE. Furthermore, a 10 km radius monitoring zone (Fig. 3) was
set up.​
Figure 3. LPAI H7N3 in the Province of Venice: location of the infected farm in the municipality of
Santa Maria di Sala and the 1 km radius restriction zone and the 10 km radius monitoring zone.​
Pagina 7 di 22​
The stamping out of all the birds present at the infected farm was started and
completed on 25 May. The serological and virological tests conducted in the
restriction and monitoring zones did not reveal any circulation of the virus in the
adjacent areas.
The results of the phylogenetic analysis conducted on the strain are shown below
(Fig. 4 and 5):​
?​
no significant homology was revealed with the H7N3 LPAI virus strains that
caused the 2002-2004 epidemic in Italy and with the H7N1 LP-HPAI strains
responsible for the 1999-2001 Italian epidemic. The Santa Maria di Sala virus
strain is, in fact, part of distinct cluster clearly separated from the clusters of the
viruses that caused the two Italian epidemic.

?​
the HA gene sequence presents elevated homology (98.3%) with the sequence of
the same gene of the strain A/chicken/England/4054/2006 (H7N3).

?​
the NA gene sequence shows a 97.7% homology with a LPAI H7N3 strain
isolated in 2001 from a wild duck in Italy.

?​
the preliminary observations of the strain?s nucleotide sequences indicate its
recent introduction in domestic poultry. This hypothesis is supported by:

?​
absence of the stalk deletion in the NA protein;

?​
absence of additional glycosylation sites in the HA protein;

?​
elevated homology with strains circulating in Italian and
European wildfowl.

Pagina 8 di 22​
A/turkey/Italy/8000/2002(H7N3))
A/turkey/Italy/251/2003(H7N3)) segmen...
A/ty/It/220158/02(H7N3)) neuraminidas...
A/turkey/Italy/220158/2002(H7N3)) NA ...
A/turkey/Italy/3620/2003(H7N3)) segme...
A/turkey/Italy/2987/2003(H7N3)) segme...
A/turkey/Italy/1010/2003(H7N3)) segme...
A/turkey/Italy/214845/2002(H7N3)) NA ...
A/turkey/Italy/214845/02(H7N3)) neura...
A/turkey/Italy/8912/2002(H7N3)) segme...
A/turkey/Italy/4608/2003(H7N3)) segme...
A/quail/Italy/3347/2004(H7N3)) segmen...
A/turkey/Italy/3337/2004(H7N3)) segme...
A/turkey/Italy/4479/2004(H7N3)) segme...
A/turkey/Italy/3807/2004(H7N3)) segme...
1941 A/turkey/Italy/2043/2003(H7N3) I...
A/mallard/Italy/33/01(H7N3)) neuramin...
A/mallard/Italy/43/01(H7N3)) neuramin...
A/chicken/Italy/2837-54/2007 NA
A/mallard/Netherlands/12/2000(H7N3)) ...
A/chicken/Pakistan/34669/95(H7N3)) ne...
A/turkey/England/63(H7N3)) neuraminid...
A/turkey/England/​
100 1963(H7N3)) segment ...

98
78
73
100
94
97
100
74
43
42
53
72
38
54
31​
0.01​
(Similarity = 96.8%)​
A/ck/Italy/2837/2007​
Figure 4 - A/Chicken/Italy/2837/2007 (H7N3) LPAI ? Phylogenetic analysis of the entire HA gene
sequence.
Figure 5 - A/Chicken/Italy/2837/2007 (H7N3) LPAI ? Phylogenetic analysis of the entire NA gene
sequence.​
A/turkey/Italy/4479/2004(H7N3)) segme...
A/quail/Italy/3347/2004(H7N3)) segmen...
A/turkey/Italy/3807/2004(H7N3)) segme...
A/turkey/Italy/8912/2002(H7N3)) segme...
A/turkey/Italy/214845/2002(H7N3)HA
A/turkey/Italy/251/2003(H7N3)HA A/turkey/Italy/251/2003(H7N3)) segmen...
A/quail/Italy/4610/2003(H7N3)) segmen...
A/mallard/Italy/33/01(H7N3)) hemagglu...
A/mallard/Italy/33/01(H7N3)HA A/mallard/Italy/43/01(H7N3)) hemagglu...
A/chicken/Italy/2837-58/2007 HA
A/chicken/Italy/2837-54/2007 HA
A/chicken/England/4266/2006(H7N3)) se...
A/chicken/England/4054/2006(H7N3)) se...
A/duck/Mongolia/47/01(H7N1)) HA gene ...
A/mallard/Netherlands/12/00(H7N3)) he...
A/mallard/Netherlands/12/2000(H7N3)) ...
A/Mallard/Sweden/107/02(H7N7)) hemagg...
A/Netherlands/219/03(H7N7)) hemagglut...
A/chicken/Netherlands/1/03(H7N7)) hem...
A/Netherlands/33/03(H7N7)) hemaggluti...
A/Chicken/Germany/R28/03(H7N7)) HA ge...
A/chicken/Italy/322/2001(H7N1)) segme...
A/Chicken/Italy/1067/99(H7N1)) H7 gen...
2043 A/turkey/Italy/977/1999(H7N1) IT...
A/Turkey/Italy/13467/99(H7N1) A/Chicken/Italy/13307/99(H7N1)) A/Turkey/Italy/13468/99(H7N1) A/ostrich/Italy/1038/2000(H7N1) A/ostrich/Italy/2332/00(H7N1) A/chicken/Italy/13474/99(H7N1) A/turkey/Italy/12598/99(H7N1) A/Chicken/Italy/13489/99(H7N1) A/Chicken/Italy/267/00(H7N1) A/swan/Shimane/42/99(H7N8)) HA gene f...
A/ostrich/South Africa/M320/96(H7N7))...
A/turkey/Ireland/PV74/1995(H7N7)) hem...
A/England/268/1996(H7N7)) hemagglutin...
A/Mallard/Sweden/56/02(H7N7)) hemaggl...
A/ostrich/South Africa/1069/91(H7N1))... A/ostrich/Zimbabwe/222/96(H7N1)) hema... A/duck/Nanchang/1904/1992(H7N1)) segm...
A/fairy bluebird/Singapore/F92/94(H7N... A/conure/England/766/94(H7N1)) hemagg...
A/parakeet/Netherlands/267497/94(H7N1...
A/common iora /Singapore/F89/95(H7N1)...
A/conure/England/1234/94(H7N1)) hemag...
A/parrot/England/1174/94(H7N1)) hemag...
A/chicken/Pakistan/CR2/95(H7N3)) hema...
A/chicken/Pakistan/447/95(H7N3)) hema...
A/chicken/Pakistan/34669/1995(H7N3)) ...
A/chicken/Hebei/1/2002(H7N2)) hemaggl...
A/psittacine/Italy/1/91(H7N2)) hemagg... A/gull/Italy/692-2/93(H7N2)) hemagglu... A/duck/Taiwan/33/1993(H7N7)) HA gene ...​
9
10
10
10
9
4
7
6
10
7
10
9
9
5 8
7
10
9
9
6
10
10
8
10
10
9
8
5
8
9
9
6
9
9
10
7
6
5
7
9
9
8
8
6
6
10​
0.02​
A/ck/Italy /2837/2007​
Strain of the Italian H7N3 LPAI epidemic (2001-2004)
Strain of the Italian H7N1 LP/HPAI epidemic (1999-2001)​
Pagina 9 di 22​
The FIESSO UMBERTIANO (RO) LPAI H7N3 outbreak​
Tests carried out in the frame of the national avian influenza monitoring plan at an
ornamental bird/dealer farm in the municipality of Fiesso Umbertiano (RO) revealed
a positivity for a LPAI H7N3 virus that was confirmed on 19 July 2007. The birds did
not present any symptoms at the moment of sampling. The Veneto Region issued
eradication measures as specified by EU Directive 94/2005/CE. Furthermore, a 10
km radius monitoring zone (Fig. 6) was set up. On 20/07/07, 2114 birds were
stamped out; 25 birds of endangered species were not suppressed .​
Figure 6. LPAI H7N3 in the Province of Rovigo: location of the infected farm in the municipality of
Fiesso Umbertiano and the 1 km radius restriction zone and the 10 km radius monitoring zone.​
Pagina 10 di 22​
THE BASILICATA REGION OUTBREAK
The H7 AI ? RUOTI (PZ) outbreak​
On 31 July 2007, serological positivity for AI sub-type H7 was diagnosed during
national avian influenza monitoring activities. The birds at the family-run farm did
not show any clinical symptoms at the moment of sampling. The farm located in
Molino Grosso in Agro in the municipality of Ruoti (PZ) (Fig. 7) contained 31
chickens, 5 turkeys, 3 guinea fowls, 21 ducks and 15 geese. The Basilicata Region
issued eradication measures as specified by EU Directive 94/2005/CE and on 9
August 2007 all the birds present were killed and destroyed.​
Figure 7 ? LPAI H7 in the Province of Potenza: location of the infected back-yard flock in the
municipality of Ruoti.​
Pagina 11 di 22​
THE EMILIA ROMAGNA REGION OUTBREAKS​
Three outbreaks of LPAI have been confirmed in the Emilia Romagna Region since
14 August, 2007 (Fig. 8). The respective data are summarised in Table 3.​
Table 3 ? LPAI outbreaks in the Emilia Romagna Region confirmed as of 10/09/07.​
Date of
confirmation
Subtype Diagnosis Municipality No. of birds
14 August 2007 H5N2 LPAI Virological
Lugo di
Romagna (RA)
10.150
17 August 2007 H7
Serological
Virological
Conselice (RA) 215
21 August 2007 H7
Serological
Virological
Lugo di
Romagna (RA)
7.100​
Figure 8. Location of LPAI outbreaks in the Emilia Romagna Region.​
Pagina 12 di 22​
The LUGO DI ROMAGNA (RA) H5N2 LPAI outbreak​
As part of the national AI monitoring plan, a free range mallard and geese breeding
farm located in Lugo (RA) was subjected to sampling on 9 August, 2007. At the
moment of sampling, 7000 mallards (born on the farm in the April-June period), 3000
geese (of which 2000 were raised in a second farm located 1.5 km from the main
farm from which they were moved on 29 July 2007) and 150 chickens for selfconsumption
were present at the farm. None of these birds showed symptoms at the
moment of sampling. On 14 August 2007, the National Reference Laboratory for
Avian Influenza (IZS delle Venezie) confirmed the isolation of a Type A, sub-Type
H5N2 LPAI virus in cloacal swabs taken from mallards (deduced amino acid
sequence at the cleavage site of the hemagglutinin: -PQRETRGL*F).​
Figure 9. Emilia Romagna Region LPAI outbreaks: restriction and monitoring zones.​
The preliminary results of the phylogenetic analysis of the HA gene showed elevated
homologies with strains isolated in 2005 from a duck in Bavaria
(A/H5N2/mallard/Bavaria/1/2005) and from a teal in Italy (A/H5N2/teal/Italy/3931-
38/2005) of 98.5% and 98.4% respectively. This strain also showed low homology​
Pagina 13 di 22​
(93.2%) with the strain (A/H5N2/turkey/Italy/1325/2005) in circulation during Italy?s
H5N2 epidemic in 2005 (Fig. 10). The Emilia Romagna Region issued eradication
measures as specified by EU Directive 94/2005/CE. Furthermore, a 3 km radius
restriction zone and a 10 km radius monitoring zone were set up around both farms
(Fig. 9). Between 15-16 August, 2007, all the birds at the farm and its functionally
linked structures (2000 geese raised at the farm 1.5 km away) were stamped out and
destroyed.​
gA/teal/Italy/3931-38/2005(H5N2)) seg...
A/teal/Italy/3931/2005(H5N2)) haemmag...
A/mallard/Bavaria/1/2005(H5N2)) hemag...
A/duck/Italy/4445/2007 (H5N2) HA
A/teal/Italy/3812/2005(H5N3)) segment...
gA/duck/Italy/775/2004(H5N3)) segment...
A/duck/France/05054a/2005(H5N3)) HA g...
A/duck/France/05057b/2005(H5N2)) HA g...
A/mallard/Netherlands/3/99(H5N2)) hem...
A/duck/Denmark/65047/04(H5N2)) hemagg...
A/mallard/Sw eden/39/02(H5N3)) hemaggl...
A/mallard/Sw eden/2/02(H5N2)) hemagglu...
A/mallard/Sw eden/7/02(H5N2)) hemagglu...
A/Chicken/Italy/367/97) segment 4 hem...
A/poultry/Italy/367/1997(H5N2)) segme...
A/poultry/Italy/373/1997(H5N2)) segme...
A/chicken/Italy/312/1997(H5N2)) segme...
A/mallard/Italy/36/2002(H5N3)) hemagg...
A/migratory duck/Hong Kong/MP206/2004...
gA/duck/Mongolia/54/01(H5N2)) HA mRNA...
A/mallard/Sw eden/58/02(H5N3)) hemaggl...
A/chicken/Italy/8/1998(H5N2)) hemaggl...
A/mallard/Italy/208/2000(H5N3)) hemag...
A/chicken/France/03426a/03(H5N2)) HA ...
A/turkey/Italy/1258/2005(H5N2)) segme...
A/turkey/Italy/1325/2005(H5N2)) segme...
A/cygnus olor/Italy/742/2006(H5N1)) s...
A/cygnus olor/Croatia/1/2005(H5N1)) s...
A/mallard/Italy/835/2006(H5N1)) segme...
A/mallard/Bavaria/1/2006(H5N1)) hemag...
A/turkey/Italy/1980(H5N2)) segment 4 ...
A/duck/Hong Kong/342/1978(H5N2)) hema...
A/duck/France/02166/02(H5N3)) HA gene...
gA/chicken/Ibaraki/1/2005(H5N2)) HA g...
A/chicken/Ibaraki/​
100 2/2005(H5N2)) HA ge...

94
87
100
100
100
99
94
85
52
100
93
96
47
95
94
87
76
88
37
100
100
97
99
77
77
88
99
100
88
49
37
0 .02​
Figure 10. Phylogenetic analysis of the nucleotide sequence of the gene that codes for the
hemagglutinin of the strain A/H5N2/duck/Italy/4445/2007 isolated in Lugo di Romagna (RA).​
Pagina 14 di 22​
The CONSELICE (RA) H7 LPAI outbreak​
On 13 August 2007, birds at a small dealer farm that sold broilers, guinea fowl, layers
and geese located in Conselice (RA) tested serologically positive for AI, subtype
(confirmation was made on 17 August 2007). None of the farm?s 60 geese, 150
guinea fowl, or 5 layers showed any symptoms at the moment of sampling. The
guinea fowl (17/20 serums) and the hens tested positively serologically, while
virological tests provided negative outcome. The Emilia Romagna Region issued
eradication measures as specified by EU Directive 94/2005/CE. Furthermore, a 10
km radius monitoring zone (Fig. 9) was set up, and on 19 August 2007 all the farm?s
birds were stamped out and destroyed.​
The LUGO DI ROMAGNA (RA) H7 AI outbreak​
A check conducted on 18 August 2007 at a dealer farm with 6.600 broilers and 500
guinea fowl located in the monitoring zone set up around the previous outbreak at
Lugo tested serologically positive for AI, subtype H7 (confirmation was made on 21
August 2007). The Emilia Romagna Region issued eradication measures as specified
by EU Directive 94/2005/CE and a 10 km radius monitoring zone (Fig. 9) was set up.
None of the birds showed clinical symptoms at the moment of sampling. All the birds
were stamped out and destroyed during the night of 23 and 24 August.
Epidemiological investigation revealed that the owner of the farm had commercial
links with the supplier of the guinea fowl that tested positive in outbreak 2.​
Pagina 15 di 22​
THE LOMBARDIA REGION OUTBREAKS​
As of 10/09/2007, no. 7 outbreaks of Avian Influenza had been confirmed in the
Lombardia Region (Tab. 4). The geographical distribution of LPAI outbreaks in
Lombardia is illustrated in figure 11.​
Table 4 ? LPAI Outbreaks in the Lombardia Region confirmed as of 10/09/07.​
Date of
confirmation
Subtype Diagnosis Municipality No. of birds
22 August 2007 H7N3
Serological and
Virological
Paderno
Franciacorta (BS)
11.400
22 August 2007 H7
Serological and
Virological
Paderno
Franciacorta (BS)
15.860
24 August 2007 H7 Serological Filago (BG) 1.446
27 August 2007 H7
Serological and
Virological
Volongo (CR) 7.898
31 August 2007 H7
Serological and
Virological
Coccaglio (BS) 11.697
3 September 2007 H7N3 Virological Rogno (BG) 4.157
7 September 2007 H7
Serological and
Virological
Castel Goffredo
(MN)
22.000​
Pagina 16 di 22​
Figure 11. Location of LPAI outbreaks in the Lombardia Region.​
Pagina 17 di 22​
The PADERNO FRANCIACORTA (BS) H7N3 LPAI and H7 LPAI outbreaks​
On 21 August 2007, a veterinarian working in the poultry production chain reported a
suspected outbreak of avian influenza in a 100 day-old meat turkey flock located in
Paderno Franciacorta (BS) on the basis of symptoms observed. The birds presented
respiratory symptoms accompanied by loss of appetite and a slightly increased
mortality rate (70 turkeys/day out of 11.400 males present). On 22 August 2007,
serological positivity for subtype H7 (1:16-1:256) was confirmed. An H7N3 avian
influenza virus​
(A/turkey/Italy/4527/2007) was subsequently isolated in this flock.
This isolate was characterized as LPAI with an intravenous pathogenicity (IVPI)
index of 0.00
. On 22 August 2007, virological and serological tests were conducted at
another meat turkey farm (with 15.860 males) owned by the same owner in the
immediate vicinity (around 150 meters distant). These birds tested positive for avian
influenza virus H7 (39/40 positive serums with variable titres from 1:16 to 1:256).
RT-PCR provided a negative outcome. The Lombardia Region issued eradication
measures as specified by EU Directive 94/2005/CE and a 10 km radius monitoring
zone was set up. All the birds in the first and in the second outbreaks were stamped
out and destroyed, on 23 and 24 August respectively.
Phylogenetic analysis based on the complete sequence of the HA gene
(A/turkey/Italy/4527/2007
- Fig. 12) demonstrated that the virus in question had an
extremely high homology (99.8%) with the virus (A/chicken/Italy/3981-90/2007-
H7N3) isolated in July from LPAI affected birds in the oubreak of Fiesso
Umbertiano, Province of Rovigo.
The virus A/turkey/Italy/4527/2007(H7N3) isolated in the Brescia area was not
related to the virus isolated in chickens in May, 2007 (S.Maria di Sala ? VE; 94.6%
homology) or with the viruses recently isolated (2006-2007) from wild geese in Italy.
The nucleotide sequences of the viruses A/chicken/Italy/3981-90/2007 (H7N3) and
A/turkey/Italy/4527/2007(H7N3) showed an average homology of 96% with the
viruses responsible for the 2002 LPAI H7N3 epidemic and for such reason could not
be closely correlated while indicating a mean homology of 93.9% with the virus

Pagina 18 di 22​
strains responsible for the 1999-2001 LPAI and HPAI H7N1 epidemic. Analysis of
the HA sequence of the 2 viruses (3981-90 and 4527) showed classic glycosylation
sites without demonstrating the presence of additional glycosylation sites in position
123 and 149. The aggregate data leads to the supposition of a still limited circulation
of the viruses in domestic poultry. The analysis of the NA protein coding gene and
the other gene segments for the virus A/turkey/Italy/4527/2007(H7N3) is still in
progress.​
Pagina 19 di 22​
A /q u a il/Ita ly /3347/2004(H7N3) ) s egme n ...
A /turkey /Italy /3337/2004(H7N3) ) s egme ...
A /turkey /Italy /4479/2004(H7N3) ) s egme ...
A /turkey /Italy /3807/2004(H7N3) ) s egme ...
A /turkey /Italy /4608/2003(H7N3) ) s egme ...
A /turkey /Italy /3620/2003(H7N3) ) s egme ...
A /turkey /Italy /2987/2003(H7N3) ) s egme ...
A /turkey /Italy /1010/2003(H7N3) ) s egme ...
A /Ty /It/9742/02 H7N3
A /turkey /Italy /8912/2002(H7N3)HA
A /turkey /Italy /8000/2002 (H7N3 )
A /turkey /Italy /21484 5 /0 2 (H7N3)HA
A /turkey /Italy /22015 8 /2002(H7N3)HA
A /turkey /Italy /251/2003(H7N3)HA
A /turkey /Italy /2043/2003(H7N3) I...
A /c hic ke n /Ita ly /3981-9 0 /2 0 0 7 (H7N3 ) HA
A /turkey /Italy /4527/2 0 0 7 HA
A /malla rd /Ita ly /3 3 /0 1 (H7N3)HA
A /malla rd /Ita ly /4 3 /0 1 (H7N3) ) hema g g lu ...
A /mallard/Italy /4234/2004(H7N7) ) hema ...
A /Mallard/Italy /497-2 9 /2 0 0 6 H7N7 (HA ...
A /Shov e ler /Italy /377-6 /2006 H7N7 (HA ...
A /Mallard/Italy /2699-7 8 /2006 H7N7 (HA ...
A /duc k/Ita ly /4692-3 0 /2004(H7N7) ) hema ...
A /mallard/Italy /4810-7 /2004(H7N7) ) h ...
A /mallard/Italy /4810-7 9 /2004(H7N4) ) h ...
A /c hic ke n /Ita ly /2837-5 8 /2 0 0 7 (H7N3 ) HA
A /c hic ke n /Ita ly /2837-5 4 /2 0 0 7 (H7N3 ) HA
A /Shov e le r/Italy /2698-2 7 /2006 H7N7 (H...
A /c hic ke n /En g land/4266/2006(H7N3) ) s e...
A /c hic ken/En g land/4054/2006(H7N3) ) s e...
A /mallard/Ne th e rlands /1 2 /0 0 (H7N3) ) h e ...
A /c hic ke n /Neth e rlands /1/0 3 (H7N7) hema ...
A /turkey /Italy /1351/2001(H7N1) ) s egme ...
A /c hic ke n /Ita ly /322/2001(H7N1) ) s egme ...
A /turkey /Italy /4426/2000(H7N1) ) s egme ...
A /c hic ke n /Ita ly /1082/1999(H7N1) ...
A /turkey /Italy /3775/9 9 (H7N1 )) HA g e n ...
A /c hic ke n /Ita ly /2335/2000 (H7N1 )
A /os tr ic h /Ita ly /2332/0 0 (H7N1) ) s egme n ...
A /c hic ke n /Ita ly /4746/1999(H7N1) ...
A /q u a il/Ita ly /396/2000(H7N1) )
A /g u ll/Ita ly /692-2 /9 3 (H7N2) ) h ema g g lu ...
A /c hic ke n /Pakis ta n /CR2 /9 5 (H7N3) ) hema ...
A /c hic ke n /Pakis ta n /447/9 5 (H7N3) ) hema ...
A /c hic ke n /​
6 4 Pakis ta n /34669/1995(H7N3) ) ...

1 0 0
1 0 0
1 0 0
9 9
1 0 0
6 4
4 4
9 7
1 0 0
4 5
3 8
9 0
9 0
1 0 0
1 0 0
8 1
6 5
9 9
5 5
9 8
8 3
6 2
8 0
9 9
9 6
3 1
9 4
1 4
4 0
8 9
7 0
8 2
8 3
8 9
6 0
5 1
4 8
6 8
8 0
5 8
0 .0 1​
Figure 12. Phylogenetic tree based on the complete sequence of the HA gene, subtype H7. The
most pertinent sequences are marked with a black ring.​
Pagina 20 di 22​
The FILAGO (BG) H7 LPAI outbreak​
A serological positivity (titres variable from 1:16 to 1:128) for avian influenza virus
H7 was confirmed on 24 August in turkeys on a farm in Filago (BG) with 1.446 birds
of various species including ornamental birds. On 3 September, all the animals were
killed and the carcases disposed of. The origin of the outbreak was related to the
introduction (on 11 and 18 July 2007) of live poultry from a previously LPAI
affected dealer farm at Lugo di Romagna (outbreak No. 6)​
The VOLONGO (CR) H7 LPAI outbreak​
On 24 August 2007, an AI outbreak was reported in a meat turkey holding with 8.560
females in Volongo (CR). The birds presented early respiratory symptoms (beginning
on 18 August 2007) and slightly increased mortality rate (approx. 1.5%). On 27
August, seropositivity for subtype H7 (1:16-1:128) was confirmed. At the moment of
seizure, there were 7.898 live birds on the farm. H7 positivity was confirmed on 29
August 2007 through RT-PCR analysis of tracheal swabs. The isolation of the virus is
currently in progress. On 31 August, all the animals were killed and the carcases
disposed of.​
The COCCAGLIO (BS) H7 LPAI outbreak​
Another AI outbreak was confirmed on 31 August 2007 in an industrial farm of
11.697 meat turkeys (males) located in Coccaglio (BS). Approximately 50% of the
birds present at the moment of confirmation (5800) presented early respiratory
symptoms (beginning on 14 August 2007). This was accompanied by an increase in
the mortality rate (around 6% of the total number of birds). Serological tests (HI)
demonstrated H7 subtype positivity with antibody titres variable from 1:16 to 1:256.
RT-PCR analysis conducted on a pool of tracheal swabs confirmed positivity for
avian influenza Type A subtype H7 on 30 August. Further analysis is currently being
conducted. Stamping out was completed on 3 September 2007.​
Pagina 21 di 22​
The ROGNO (BG) H7 LPAI outbreak​
An outbreak of H7 LPAI was confirmed on 3 September 2007 in a dealer farm at
Rogno (BG). At the moment of seizure, there were 4.157 live birds on the farm:
mainly laying hens, broilers, capons, meat turkeys, guinea fowl, ducks, pheasant,
partridges and grey partridges. None of the birds showed respiratory symptoms at the
moment of sampling. A H7N3 virus was isolated from samples taken from the
affected birds. Stamping out has been completed on 6 September 2007. Phylogenetic
analysis based on the complete sequence of the HA gene (A/duck/Italy/4936/2007 ?
Fig. 13) demonstrated that the virus in question has a 100% homology with the virus
A/turkey/Italy/4527/2007(H7N3) isolated in August from affected birds in the LPAI
outbreak of Paderno Franciacorta, Province of Brescia.​
The CASTEL GOFFREDO (MN) H7 LPAI outbreak​
An outbreak of H7 LPAI was confirmed on 7 September 2007 in a dealer farm at
Castel Goffredo (MN). At the moment of seizure, there were 22.000 live birds on the
farm: broilers, capons, ducks and geese. None of the birds showed respiratory
symptoms at the moment of sampling. A H7 positivity was obtained through RT-PCR
analysis of faecal samples taken from geese and ducks. This outbreak was
epidemiologically linked to a previously LPAI affected dealer farm at Rogno
(outbreak No. 12): both producers exchanged birds at the Montichiari market in the
province of Brescia.​
Pagina 22 di 22

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