Integration of genetic and epidemiological data to infer H5N8 HPAI virus transmission dynamics during the 2016-2017 epidemic in Italy.

Between October 2016 and December 2017, several European Countries had been involved in a massive Highly Pathogenic Avian Influenza (HPAI) epidemic sustained by H5N8 subtype virus. Starting on December 2016, also Italy was affected by H5N8 HPAI virus, with cases occurring in two epidemic waves: the first between December 2016 and May 2017, and the second in July-December 2017. Eighty-three outbreaks were recorded in poultry, 67 of which (80.72%) occurring in the second wave. A total of 14 cases were reported in wild birds. Epidemiological information and genetic analyses were conjointly used to get insight on the spread dynamics. Analyses indicated multiple introductions from wild birds to the poultry sector in the first epidemic wave, and noteworthy lateral spread from October 2017 in a limited geographical area with high poultry densities. Turkeys, layers and backyards were the mainly affected types of poultry production. Two genetic sub-groups were detected in the second wave in non-overlapping geographical areas, leading to speculate on the involvement of different wild bird populations. The integration of epidemiological data and genetic analyses allowed to unravel the transmission dynamics of H5N8 virus in Italy, and could be exploited to timely support in implementing tailored control measures.

The economic implications of sylvatic rabies eradication in Italy.

After more than 10 years of absence, sylvatic rabies re-appeared in Italy in 2008. To prevent disease spread, three oral rabies vaccination (ORV) campaigns targeting red foxes were performed through manual distribution of vaccine baits between January and September 2009. As these campaigns proved unsuccessful, at the end of December 2009, baits started being distributed using helicopters, allowing uniform coverage of larger areas in a shorter time period. From winter 2009 to autumn 2016, a total of 15 ORV campaigns (four emergency, four regular and seven preventive ORV) were implemented through aerial distribution of baits. In this study, we assessed the costs of the aerial ORV campaigns, which were aimed at eradicating the disease and reobtaining the rabies-free status. Cumulative costs per km2 were estimated at €59.45 during emergency campaigns and ranged between €51.94 and €65.67 in the regular vaccinations. The main portion of costs for ORV programmes were related to baits supply and distribution: €49.24 (82.83%) in emergency campaigns and from €40.33 to € 43.35 in regular ORVs (71.97% and 66.02%, respectively). At the end of each ORV campaign, the efficacy of vaccination activities was estimated by assessing the proportion of foxes testing positive for tetracycline biomarker in jawbone, indicating bait intake. Results revealed that the proportion of foxes that ingested baits varied between 70.97% and 95.51%. Statistical analysis indicated that reducing the density of dropped baits could potentially lead to a cost-saving of 22.81%, still maintaining a satisfactory level of bait intake by the fox population.

H7N7 Highly Pathogenic Avian Influenza in Poultry Farms in Italy in 2016.

After the H7N7 highly pathogenic (HP) avian influenza (AI) outbreak in 2013, and a single case of H5N8 HPAI in 2014, in April 2016, a H7N7 HPAI virus was detected in northeastern Italy. The case occurred in an organic free-range laying hen farm located in proximity with one of the highest densely populated poultry areas (DPPAs) in Italy. Control measures provided by the Council of the European Union in directive 2005/94/CE were promptly applied, and enhanced surveillance activities were implemented in the DPPAs. On May 16, 2016, a second case was confirmed in a fattening turkey farm within the protection zone of the previous outbreak. Following an epidemiologic inquiry, another turkey farm was considered at risk of transmission and was subjected to preemptive culling. Epidemiologic data and phylogenetic analyses indicated that the virus was likely introduced from wild birds as a low pathogenicity AI strain, through direct contact. The rapid containment of the outbreak proves the level of preparedness of the veterinary public health sector in Italy. Nevertheless, the recurrent introductions from wild birds indicate the need of improving both the biosecurity levels in the DPPA and the surveillance activities in wild birds to quickly detect the presence of AI in the territory.

Factors associated with pre-slaughter mortality in turkeys and end of lay hens.

Pre-slaughter transportation may affect poultry welfare and mortality rates. A retrospective analysis was conducted to examine the effect of environmental, management and individual factors on the percentage of dead birds during pre-slaughter transportation (dead-on-arrival, DOA). The variables accounted for in the analyses included: environmental temperature, travel duration, genetic line, gender, crate type and crate stocking density. Among the 41 452 loads of turkeys (34 696 388 birds) and 3241 of end of lay hens (21 788 124 birds) transported to three large abattoirs in northern Italy in a 3-year period, the median DOA was 0.14% in turkeys, and 0.38% in hens. In turkeys, travel duration longer than 30 min, temperature higher than 26°C and high in-crate densities were associated with increased DOA. In winter (⩽2°C), high stocking densities did not reduce the mortality risk from cold stress; on the contrary, for stocking densities either near to or just above the maximum density in EC Reg. 1/2005, the DOA risk was greater than for loads with densities of 10 kg/m2 less than the EC maximum. Male birds and specific genetic lines also showed a higher DOA. In hens, transportation lasting longer than 2 h and the brown-feathered breed were associated with higher DOA. Dead-on-arrival progressively increased with travel duration, remaining constant between 4 and 6 h and peaking at 8 h (median: 0.57%). The maximum DOA increase was detected during winter. These results show that several species-specific factors may lead to increased risk of mortality.

Retrospective space-time analysis methods to support West Nile virus surveillance activities.

The steep increase in human West Nile virus (WNV) infections in 2011-2012 in north-eastern Italy prompted a refinement of the surveillance plan. Data from the 2010-2012 surveillance activities on mosquitoes, equines, and humans were analysed through Bernoulli space-time scan statistics, to detect the presence of recurrent WNV infection hotspots. Linear models were fit to detect the possible relationships between WNV occurrence in humans and its activity in mosquitoes. Clusters were detected for all of the hosts, defining a limited area on which to focus surveillance and promptly identify WNV reactivation. Positive relationships were identified between WNV in humans and in mosquitoes; although it was not possible to define precise spatial and temporal scales at which entomological surveillance could predict the increasing risk of human infections. This stresses the necessity to improve entomological surveillance by increasing both the density of trapping sites and the frequency of captures.

Emergency oral rabies vaccination of foxes in Italy in 2009-2010: identification of residual rabies foci at higher altitudes in the Alps.

Following a resurgence of fox rabies in northeastern Italy in 2008-2009, two emergency oral rabies vaccination (ORV) campaigns were performed in the Alpine mountain ranges in 2009 and 2010 using aerial distribution to prevent the disease from spreading further inland. Vaccine baits were distributed only below the freezing point altitude, 1000 m above sea level (a.s.l.) in December 2009-January 2010 and 1500 m a.s.l. in April-May 2010, to avoid repeated freeze-thaw cycles. Spatial analysis unexpectedly identified fox rabies hotspots above the threshold altitudes, probably representing local residual rabies foci which may have contributed to maintaining the infectious cycle in areas not vaccinated at higher altitudes. Based on the results obtained, in May 2010, the second ORV campaign was extended to include threshold altitudes of up to 2300 m a.s.l. to eliminate residual foci. The observations made may help in the formulation of ORV strategies in countries sharing similar topographical features.

Impact of emergency oral rabies vaccination of foxes in northeastern Italy, 28 December 2009-20 January 2010: preliminary evaluation.

Fox rabies re-emerged in northeastern Italy in 2008, in an area bordering Slovenia. In 2009, the infection spread westward to Veneto region and in 2010 to the provinces of Trento and Bolzano. Aerial emergency oral fox vaccination was implemented in the winter 2009-10. Since this vaccination was performed at altitudes below the freezing level, a statistical analysis was conducted to evaluate its impact. Of the foxes sampled following the vaccination campaign, 77% showed a rabies antibody titre of >or=0.5 IU/ml.

Field evidence of the efficacy of vaccination to control low pathogenicity avian influenza in meat turkeys.

This paper analyzes the efficacy of vaccination to control low pathogenicity avian influenza outbreaks using information collected during four epidemics occurring in Italy between 2000 and 2005. Different vaccination strategies and protocols for meat-turkey immunization are also considered.

Modelling the spatial spread of H7N1 avian influenza virus among poultry farms in Italy.

We analysed the between-farm transmission of the H7N1 highly pathogenic avian influenza virus that disrupted the Italian poultry production in the 1999-2000 epidemic with a SEIR model with a spatial transmission kernel, accounting for the containment measures actually undertaken. We found significant differences in susceptibility between species and a reduction in transmissibility after the first phase. We performed simulations to assess the effectiveness of the implemented and new control measures. The most effective measure was the ban on restocking. An earlier start of pre-emptive culling promotes eradication; restricted pre-emptive culling delays eradication but causes lower losses.

Evaluation of interventions and vaccination strategies for low pathogenicity avian influenza: spatial and space-time analyses and quantification of the spread of infection.

In recent years the control of low pathogenicity avian influenza (LPAI) viruses of the H5 and H7 subtypes has increasingly become a concern. We evaluated the measures (stamping out, controlled marketing, emergency and preventive vaccination, farm density reduction and restocking in homogenous areas) implemented to control the LPAI epidemics that occurred in Italy between 2000 and 2005, using a combination of spatial and space-time analyses and estimates of the basic reproduction ratio (R0). Clustering of infected farms decreased over the years, indicating the effectiveness of the control strategies implemented. Controlled marketing [relative risk (RR) 0.46, 95% confidence interval (CI) 0.27-0.80], emergency (RR 0.47, 95% CI 0.39-0.57) and preventive vaccination (RR 0.19, 95% CI 0.09-0.41) were the most effective measures, yet R0<1 was only for preventive vaccination. Our results are useful for identifying the most effective measures for reducing the risk of the spread of LPAI and optimizing the allocation of resources.

Spatial analysis of the 1999-2000 highly pathogenic avian influenza (H7N1) epidemic in northern Italy.

The effect of proximity on infected premises was evaluated during the highly pathogenic avian influenza (HPAI) epidemic that struck northern Italy in 1999-2000 by quantifying the spatial and temporal clustering of cases. The epidemic was caused by an H7N1 subtype of type A influenza virus that originated from a low-pathogenic AI virus that spread among poultry farms in northeastern Italy in 1999 and eventually became virulent by mutation. More than 90% of 413 infected premises were located in Lombardy and Veneto regions; of 382 outbreaks, 60% occurred in the Lombardy region and 40% in the Veneto region. Global and local spatial statistics were used to estimate the location and degree of clustering of cases with respect to the population at risk. Outbreaks were spatially clustered primarily in Lombardy, with a large cluster in Brescia province and another in Mantua province, on the border of Veneto. Time series analysis was used to assess the temporal clustering of outbreaks. Temporal aggregation increased during the first 5 wk and decreased thereafter (probably as a result of eradication measures enforced in the Veneto region). Spatio-temporal clustering was assessed considering the Temporal Risk Window (TRW), the time period during which premises remain infectious and infection can spread to neighboring premises. The clustering pattern was similar to the one detected when considering spatial clustering (i.e., the larger clusters were identified in the Brescia and Mantua provinces of Lombardy). These results highlight the role of proximity in the spread of AI virus and, when considering the TRW, indicate the possible direction of virus spread.