Hepatitis E virus infection in North Italy: high seroprevalence in swine herds and increased risk for swine workers.

We determined the hepatitis E virus (HEV) seroprevalence and detection rate in commercial swine herds in Italy's utmost pig-rich area, and assessed HEV seropositivity risk in humans as a function of occupational exposure to pigs, diet, foreign travel, medical history and hunting activities. During 2011-2014, 2700 sera from 300 swine herds were tested for anti-HEV IgG. HEV RNA was searched in 959 faecal pools from HEV-seropositive herds and in liver/bile/muscle samples from 179 pigs from HEV-positive herds. A cohort study of HEV seropositivity in swine workers (n = 149) was also performed using two comparison groups of people unexposed to swine: omnivores (n = 121) and vegetarians/vegans (n = 115). Herd-level seroprevalence was 75·6% and was highest in farrow-to-feeder herds (81·6%). Twenty-six out of 105 (24·8%) herds had HEV-positive faecal samples (25 HEV-3, one HEV-4). Only one bile sample tested positive. HEV seropositivity was 12·3% in swine workers, 0·9% in omnivores and 3·0% in vegetarians/vegans. Factors significantly associated with HEV seropositivity were occupational exposure to pigs, travel to Africa and increased swine workers' age. We concluded that HEV is widespread in Italian swine herds and HEV-4 circulation is alarming given its pathogenicity, with those occupationally exposed to pigs being at increased risk of HEV seropositivity.

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.

Hepatitis E virus genotype 4 in a pig farm, Italy, 2013.

Zoonotic strains of hepatitis E virus (HEV) in Europe have been reported to belong to genotypes 3 and 4. In 2012 and 2013, 57 pig farms in Northern Italy that had previously resulted seropositive for HEV were surveyed for the presence of the virus, with positive samples subsequently genotyped. Hepatitis E RNA was identified in 17/57 (29·8%) seropositive farms. Phylogenetic analysis demonstrated that distinct subtypes of genotype 3 were circulating in the north-east of Italy; as well, for the first time in the Italian swine population, genotype 4 was identified and attributed to subtype d.

Application of an integrated outbreak management plan for the control of leptospirosis in dairy cattle herds.

SUMMARY: Two outbreaks of Leptospira borgpetersenii serovar Hardjo infection in dairy cattle herds were managed through the application of enhanced biosecurity measures, whole-herd antibiotic treatment and vaccination. Micro-agglutination test antibody titres were determined in paired serum samples at 3 weeks (T1: n = 125, 97% seropositivity, median 800, range 100-12 800) and 24 weeks (T2: n = 110, 88% seropositivity, median 200, range 100-6400) after vaccination and studied in relation to cows' age, herd of origin and sampling time. From T1 to T2, vaccine-elicited antibody titres decreased by 84·7% (95% CI 76·2-90·1). Consistent with increasing immunocompetence in calves (aged <12 months) and immunosenescence in adult cows (aged >36 months) associated with ageing, antibody titres correlated positively with calves' age and negatively with adult cows' age. No cow had cultivable, (histo)pathologically detectable and/or PCR-detectable leptospires in urine or kidney samples after treatment and vaccination. Vaccination together with proper biosecurity measures and chemoprophylaxis are an affordable insurance to control bovine leptospirosis.

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.

Serosurvey against H5 and H7 avian influenza viruses in Italian poultry workers.

Highly pathogenic (HP) and low pathogenic (LP) avian influenza viruses (AIVs) belonging to H5 and H7 subtypes have been found to be associated with human infection as the result of direct transmission from infected poultry. Human infections by AIVs can cause mild or subclinical disease, and serosurveys are believed to represent an important tool to identify risk of zoonotic transmission. Therefore, we sought to examine Italian poultry workers exposed during LPAI and HPAI outbreaks with the aim of assessing serologic evidence of infection with H5 and H7 AIVs. From December 2008 to June 2010 serum samples were collected from 188 poultry workers and 379 nonexposed controls in Northern Italy. The hemagglutination inhibition (HI) assay using horse red blood cells (RBCs) and a microneutralization (MN)-enzyme-linked immunosorbent assay test were used to analyze human sera for antibodies against the following H5 and H7 LPAI viruses: A/Dk/It/4445/07(H5N2); A/Ty/It/2369/09(H5N7); A/Ty/It/218-193/ 10; A/Ck/It/3775/99(H7N1); A/Ty/It/214845/03(H7N3); and A/Dk/It/332145/09(H7N3). Since previous studies identified low antibody titer to AIVs in people exposed to infected poultry, a cutoff titer of > or = 1:10 was chosen for both serologic assays. Only HI-positive results confirmed by MN assay were considered positive for presence of specific antibodies. The Fisher exact test was used to analyze differences in seroprevalence between poultry workers and control groups, with the significance level set at P < 0.05. MN results showed a proportion of H7-seropositive poultry workers (6/188, i.e., 3.2%), significantly higher than that of controls (0/379), whereas no MN-positive result was obtained against three H5 LPAI subtypes recently identified in Italy. In conclusion, the survey indicated that assessing seroprevalence can be an important tool in risk assessment and health,surveillance of poultry workers.

West Nile virus circulation in Veneto region in 2008-2009.

West Nile virus (WNV) was detected in Italy, in late summer 2008 in horses and birds in the Po valley. As a consequence, an intense WNV surveillance was implemented in that area involving Emilia-Romagna, Veneto and Lombardy. This paper presents the results of the September 2008-November 2009 surveillance on equines, mosquitoes, wild birds, dogs and cattle in Veneto. WNV was detected in equines and dogs, and, to a lesser extent in cattle and wild birds. Simultaneous circulation of Usutu virus was detected by testing wild birds found dead. Usutu virus but not WNV was also found in mosquitoes monitored during 2009. Equine practices monitoring allowed the definition of an area of WNV circulation and the 2008-2009 westward and northward spread of the infection. Although a relatively low number of human cases and a low virus circulation in vectors and birds detected in Veneto region could be considered favourable conditions for a limited risk of human exposure, it remains difficult to predict the possible evolution of the epidemiological situation.

Epidemiology and control of low pathogenicity avian influenza infections in rural poultry in Italy.

We analyzed the involvement of the rural poultry sector in outbreaks of low pathogenicity avian influenza (AI) in Italy in 2007-2009 and discuss possible measures for improving monitoring and control. A description of how the rural poultry sector is organized also is provided. Data were obtained by the AI surveillance system established in the areas affected by the outbreaks. The surveillance activities identified two H7N3 epidemics, in 2007 and 2009, both of which mainly involved the rural sector, yet these activities did not allow for the prompt eradication of the disease. Additional strategies could be adopted to avoid the persistence of AI within the rural sector, based on the regulation and control of poultry holdings at the top of the production chain.

Low pathogenicity avian influenza in Italy during 2007 and 2008: epidemiology and control.

Since 1999, the Italian poultry production system has experienced several outbreaks of avian influenza (AI), mainly located in northeastern Italy. This paper describes the low pathogenicity (LP) AI outbreaks detected during the surveillance activities implemented in 2007-08. From May to October 2007, ten rural and hobby poultry farms were infected by an LPAI virus of the H7N3 subtype. In August-October 2007, the H7N3 LPAI virus was introduced into the industrial poultry sector with the involvement of six meat turkey farms. Phylogenetic analysis of the hemagglutinin gene indicated that all but one of the H7N3 virus strains had a high level of homology (98.7%-99.8%). Furthermore, in August 2007, an LPAI H5N2 virus was identified in a free-range geese and duck breeder flock. The hemagglutinin and neuraminidase genes showed a high level of homology (99.8% and 99.9%, respectively) with H5N2 LPAI viruses isolated from mallards in July 2007 in the same area, suggesting a possible introduction from the wild reservoir. All the birds (in total 129,386) on the infected poultry farms were culled. The prompt implementation of AI control measures, including the enforcement of a targeted emergency vaccination plan, allowed the rapid eradication of infection. In 2008, three LPAI viruses (two H7N1 and one H5N1) were identified in dealer/rural farms. The surveillance activity implemented in this area allowed the prompt detection of LPAI viruses of the H5 and H7 subtypes in the rural sector, which, as observed in the 2007 epidemic, might be the source of infection for industrial poultry.

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.

The effects of control measures on the economic burden associated with epidemics of avian influenza in Italy.

In 1999, Italy experienced a devastating epidemic of high-pathogenicity avian influenza (HPAI) caused by an H7N1 virus subtype. After this epidemic, a ministerial decree was passed to implement control measures for low-pathogenicity avian influenza (LPAI) due to H5 and H7 subtypes. We investigated whether these control measures have decreased the public expenditure associated with epidemics of LPAI and HPAI by comparing the direct and consequential losses of the 1999 epidemic to the losses associated with successive epidemics. The estimated total economic burden of the epidemics was about euro650 million (euro217 million in direct losses and euro433 million in consequential losses). The 1999 epidemic accounted for most of these losses (euro507 million: euro112 million in direct losses and euro395 million in consequential losses), whereas the total economic burden for the 5 successive LPAI was euro143 million (euro105 million in direct losses and euro38 million in consequential losses). These results demonstrate that the implementation of a coordinated set of disease-control measures, which included both emergency and prophylactic vaccination, was able to reduce the overall costs associated with avian influenza epidemics. The results also show that the application of adequate LPAI control measures may limit the risk of emergence of an HPAI virus in an area with a high poultry density, allowing the complete disruption of the poultry market and its huge associated costs to be avoided.

Adult neurogenesis without germinal layers: the "atypical" cerebellum of rabbits.

Unlike non mammalian vertebrates, adult neurogenesis in mammals is detectable in highly restricted brain sites. Persistent neurogenesis is thought to depend on stem cells residing in neural stem cell niches which are remnants of the embryonic germinal layers. Local progenitors which retain some proliferative capacity have been identified in the mature brain parenchyma, yet they do not support a constitutive, 'actual' neurogenesis, but rather a 'potential' neurogenesis which does not extrinsecate fully and spontaneously in vivo. In contrast with such a view, genesis of neuronal and glial cells from local progenitors does occur in the peripuberal and adult rabbit cerebellum. This process is independent from persisting germinal layers and involves different cell populations.

Transfer of passive immunity in dairy calves: the effectiveness of providing a supplementary colostrum meal in addition to nursing from the dam.

Failed transfer of passive immunity (FTPI) in dairy calves - which is often due to the low amount of colostrum provided within a few hours after birth - remains a crucial issue. Enabling dairy calves to nurse colostrum from their dams could be useful in increasing intake and thus avoiding FTPI, but further potential effects on the health and welfare of both calves and dams should also be considered. In this study, 107 calf-dam pairs from two Italian dairy farms were alternately assigned to one of the following colostrum provision methods (CPMs): 'hand-fed method' (HFM) - the calf was separated from the dam immediately after birth and colostrum was provided by nipple-bottle (n = 50); 'nursing method' (NM) - the calf nursed colostrum from the dam for the first 12 h of life without farmer assistance (n = 30); and 'mixed method' (MM) - the nursing calf received a supplementary colostrum meal by nipple-bottle (n = 27). Serum of calves (1 to 5 days of age) and samples of their first colostrum meal were analysed by electrophoresis to assess immunoglobulin (Ig) concentration. Additionally, behavioural indicators of separation distress (calf and dam vocalisations; calf refusal of the first meal after separation; undesirable dam behaviour at milking) in the following 24 h were recorded as binary variables (Yes/No), and the health status of calves (disease occurrence and mortality) and dams (postpartum disorders and mastitis occurrence) were monitored for the first 3 months of life and 7 days after parturition, respectively. The lowest FTPI occurrence (calf serum Ig concentration <10.0 g/l) was found in the MM (11.1%) and the HFM (22.0%) compared with the NM (60.0%) (P<0.05), and the highest percentage of calves with optimal transfer of passive immunity (serum Ig concentration ≥16.0 g/l) was observed in the MM (55.6%). The lowest calf-dam separation distress was observed in the HFM (P<0.05). The highest calf disease occurrence was recorded in the HFM (64.0%) and the lowest in the NM (33.3%), with an intermediate value for the MM (44.4%) (P<0.05). No effect of the CPM was observed on dam health or calf mortality (P>0.05). The results of this study indicated that providing calves with a supplementary colostrum meal in addition to nursing from the dam (MM) is truly effective in maximizing passive immunity transfer. Anyway, specific strategies should be studied to minimise calf-dam separation distress.

Identification of a zoonotic genotype 3 hepatitis E subtype in wildlife in north-eastern Italy.

Hepatitis E virus (HEV) is an emerging zoonosis caused by a positive RNA single stranded virus of the Hepeviridae family. In developed countries, pigs and wild boars are the main reservoir for zoonotic genotypes 3 and 4. In spring 2017, for the first time HEV was detected in wild boars captured in the Regional Park of the Euganean Hills, in north-eastern Italy. Phylogenetic analysis of two complete viral genomes and seven partial ORF1 and ORF2 sequences of HEV viruses, selected from twelve positive animals, showed that the viruses grouped together within genotype 3 but clustered separately from previously identified subtypes, thus suggesting the identification of a novel genotype 3 subtype. The phylogenetic analysis of nine partial ORF2 sequences showed the closest similarity with wild boar/human viruses identified in central-northern Italy in 2012. The circulation of HEVs in this area, characterized by a vast man-made environment, an overpopulation of wild boars and >150 swine farms, should be considered in a public health perspective. Further investigations at the wild/domestic species and human interface are therefore necessary to gain a deeper understanding of HEV dynamics.

Shc signaling in differentiating neural progenitor cells.

Previously we found that the availability of ShcA adapter is maximal in neural stem cells but that it is absent in mature neurons. Here we report that ShcC, unlike ShcA, is not present in neural stem/progenitor cells, but is expressed after cessation of their division and becomes selectively enriched in mature neurons. Analyses of its activity in differentiating neural stem/progenitor cells revealed that ShcC positively affects their viability and neuronal maturation via recruitment of the PI3K-Akt-Bad pathway and persistent activation of the MAPK pathway. We suggest that the switch from ShcA to ShcC modifies the responsiveness of neural stem/progenitor cells to extracellular stimuli, generating proliferation (with ShcA) or survival/differentiation (with ShcC).

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.

Factors associated with passive immunity transfer in dairy calves: combined effect of delivery time, amount and quality of the first colostrum meal.

Despite the well-known importance of an adequate colostral immunoglobulin (Ig) transfer to calf health and survival, failed transfer of passive immunity (FTPI) remains a widespread problem in dairy farming. The aim of this study was to investigate the management factors associated with FTPI in newborn calves, evaluating particularly the combined effect of delivery time, amount and quality of the first colostrum meal. The study was conducted from March to August 2014 on 21 Italian dairy farms. Farmers were asked as first to answer a farm-level questionnaire on calf management. Blood sampling was then performed on overall 244 calves (1 to 5 days of age) born from Holstein cows, and a sample of the first colostrum meal of each calf was collected. Individual information on calves and the respective colostrum management were recorded. Serum and colostrum Ig concentrations were assessed by electrophoresis. A mixed effects multivariable logistic regression model was used to investigate the association of the variables obtained from both the management questionnaire and the individual calf data with FTPI (calf serum Ig concentration 87.6 g/l) within 1.0 h from birth. Considerable improvements are still needed about colostrum management for newborn calves in dairy farms. The results of this study will help in developing farm-specific programs for reducing the occurrence of FTPI.

Microbiological, chemical and physical quality of drinking water for commercial turkeys: a cross-sectional study.

Drinking water for poultry is not subject to particular microbiological, chemical and physical requirements, thereby representing a potential transmission route for pathogenic microorganisms and contaminants and/or becoming unsuitable for water-administered medications. This study assessed the microbiological, chemical and physical drinking water quality of 28 turkey farms in North-Eastern Italy: 14 supplied with tap water (TW) and 14 with well water (WW). Water salinity, hardness, pH, ammonia, sulphate, phosphate, nitrate, chromium, copper and iron levels were also assessed. Moreover, total bacterial count at 22°C, presence and enumeration of Enterococcus spp. and E. coli, presence of Salmonella spp. and Campylobacter spp. were quantified. A water sample was collected in winter and in summer at 3 sampling sites: the water source (A), the beginning (B) and the end (C) of the nipple line (168 samples in total). Chemical and physical quality of both TW and WW sources was mostly within the limits of TW for humans. However, high levels of hardness and iron were evidenced in both sources. In WW vs. TW, sulphate and salinity levels were significantly higher, whilst pH and nitrate levels were significantly lower. At site A, microbiological quality of WW and TW was mostly within the limit of TW for humans. However, both sources had a significantly lower microbiological quality at sites B and C. Salmonella enterica subsp. enterica serotype Kentucky was isolated only twice from WW. Campylobacter spp. were rarely isolated (3.6% of farms); however, Campylobacter spp. farm-level prevalence by real-time PCR was up to 43% for both water sources. Winter posed at higher risk than summer for Campylobacter spp. presence in water, whereas no significant associations were found with water source, site, recirculation system, and turkey age. Low salinity and high hardness were significant risk factors for C. coli and C. jejuni presence, respectively. These results show the need of improving sanitization of drinking water pipelines for commercial turkeys.

Impact of sexual maturity on the welfare of immunocastrated v. entire heavy female pigs.

During oestrus, fattening female pigs are more prone to lameness, fractures and wounds due to mounting and agonistic behaviours of penmates. This study assessed the effect of sexual maturity on the behaviour and welfare of heavy female pigs slaughtered at 36 weeks of age (180±10 kg) for dry-cured ham production. An immunocastrated control group was used for comparison. In all, 56 15-week-old female pigs, individually identifiable by back tattoos were equally distributed among four pens. All animals from two pens were subject to a three-dose immunocastration schedule at 16, 20 and 32 weeks of age. Skin lesions and behaviours were assessed at 18, 23, 28, 33 and 36 weeks of age. A blood sample was collected at 20, 24, 28 and 32 weeks of age for assessing health/stress parameters and GnRH antibodies. At slaughter, ovaries were weighed, measured and histologically examined; stomachs, carcasses and lungs were scored for lesions and a further blood sample was taken. Immunocastrated pigs did not significantly differ from controls in growth rate, feed efficiency and slaughter performances (lung score, gastric score, backfat thickness). However, they showed a lower frequency of aggressive interactions at 33 and 36 weeks, lower front lesions at 28 weeks, but higher at 30 weeks; a lower haptoglobin level at 28 weeks, a lower level of cortisol and back lesions at slaughter (36 weeks). These findings suggest a low, yet not negligible, impact of sexual maturity on the welfare of heavy female pigs.

A simulation model for low-pathogenicity avian influenza viruses in dabbling ducks in Europe.

Anseriformes are the reservoir of low-pathogenicity avian influenza viruses (LPAIV). Studies have shown a high LPAIV prevalence associated with low antibody detection in a wild duck population in northern European countries, whereas in winter areas (Mediterranean basin), low viral detection and high seroprevalence were observed. In order to gain insight into the role played by both population recruitment and migration on AIV persistence, an epidemiological model was developed. A susceptible, infectious and removed (immune or dead)-individuals model coupling population and infection dynamics was developed to simulate LPAIV circulation in dabbling ducks throughout the entire year. The transmission coefficient (beta) was calculated using the original dataset of published works, whereas dabbling duck demographic parameters were obtained from the literature. The estimated host density threshold for virus persistence is 380 susceptible individuals per day whereas the critical community size needed for maintaining the virus throughout the winter has been estimated to be about 1200 individuals. The model showed peaks of viral prevalence after nesting and during the moult period because of population recruitment and high host density, respectively. During the winter and spring periods, the viruses reach the minimal endemic level and local extinction is highly probable because of stochastic phenomena, respectively 80% and 90% of probabilities. The most sensitive parameters of the model are the recruitment rate of young susceptible animals and the duration of virus shedding.