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The natural transmission cycle of West Nile virus (WNV) involves birds as primary hosts and mosquitoes as vectors, but this virus can spread to mammals, human beings included. Asymptomatic infected donors pose a risk to the safety of blood transfusions and organ transplants, as WNV can be transmitted through these medical procedures. Since 2009, the region of Emilia-Romagna in northern Italy has been implementing an integrated surveillance system in order to detect WNV circulation in the environment at an early stage. Here we report the results of the two components of the surveillance system, the active testing of corvids and humans, and demonstrate that bird surveillance alone improves a surveillance system based solely on human case detection. As WNV risk reduction measures are applied on a provincial basis, we assessed the ability of this surveillance system component to detect virus circulation prior to the notification of the first human case for each province. Overall, 99 epidemic seasons were evaluated as a result of 11 years (2013–2023) of surveillance in the nine provinces of the region. In this period, 22,314 corvids were tested for WNV and 642 (2.9%) were found to be infected. WNV was generally first detected in birds in July, with sample prevalence peaks occurring between August and September. During the same period, 469 autochthonous human cases were notified, about 60% of which were reported in August. WNV was detected 79 times out of the 99 seasons considered. The virus was notified in birds 73 times (92.4%) and 60 times (75.9%) in humans. WNV was first or only notified in birds in 57 seasons (72.1%), while it was first or only notified in humans in 22 seasons (27.8%). Active surveillance in corvids generally allows the detection of WNV before the onset of human cases. Failure of virus detection occurred mainly in seasons where the number of birds tested was low. Our results show that active testing of a minimum of 3.8 corvids per 100 km2 provides a satisfactory timeliness in the virus detection, but for early detection of WNV it is crucial to test birds between mid-June and mid-August.

This study analyzed data from 6 years (2014–2019) of official controls in the Emilia-Romagna region (northern Italy) to investigate the frequencies of human pathogens and chemical hazards in foods during production and distribution. Campylobacter spp. was the most prevalent pathogen, isolated in 4.4% of the 1,078 food samples examined, followed by Salmonella spp. (2.8%), Shiga toxin-producing Escherichia coli (STEC) (1.9%), and Listeria monocytogenes (0.9%). Salmonella serotyping showed that the isolates belonged to the serotypes most commonly isolated from humans in Emilia-Romagna. These serotypes were as follows: S. Infantis (34.8%), mostly isolated from chicken, monophasic S. Typhimurium (1,4, [5],12:i:-) (12.6%), S. Bredeney (8.9%), and S. Derby (8.6%). No Clostridium botulinum, Yersinia spp., and Shigella spp. were isolated. No positivity was detected for hepatitis A virus, while 5.1% of samples taken in the production phase of the food chain were found to be contaminated with norovirus. The chemical analyses identified environmental contaminants within legal limits (heavy metals, 0.6% positive overall; mycotoxins, 0.4% positive overall), analytes subjected to monitoring (perfluoro-alkyl substances (PFASs), 6.2% positive overall; inorganic arsenic, no positives overall) and process contaminants and additives within legal limits (acrylamide, 9.6% positive overall; permitted or nonpermitted additives, 0.9% positive overall). Only one sample showed dioxins and polychlorinated biphenyls (PCBs) at levels higher than the legal limits. The monitoring by competent authorities (CA) of food contamination can generate useful data that can be used as a basis for estimating the exposure to different food contaminants over time and for evaluating the effects of control measures on the contamination of food.

Paratuberculosis is a notable infectious disease of ruminants. Goats appear to be particularly susceptible. The survey aimed to investigate the spread of paratuberculosis in Italian goat farming and evaluate whether the presence of the disease could be influenced by welfare and biosecurity deficiencies. A serological survey for paratuberculosis in 33 dairy farms in northern Italy was conducted. Contextually, animal welfare and biosecurity were assessed, using a standardized protocol of 36 welfare indicators and 15 biosecurity indicators which assigns to each farm a welfare and biosecurity score from 0 (any application) to 100% (full application). An overall result of less than 60% was considered insufficient. Nineteen farms (58%) tested positive for paratuberculosis, with a mean intra-herd seroprevalence of 7.4%. Total welfare ranged from 39.56 to 90.7% (mean 68.64%). Biosecurity scores ranged from 10.04 to 90.01% (mean 57.57%). Eight farms (24%) showed poor welfare conditions (welfare score < 60%) and 19 (58%) an unsatisfactory biosecurity condition (biosecurity score < 60%). With respect to the explorative character of the study, an indicative association between seven welfare and biosecurity indicators and paratuberculosis seropositivity was identified. The presence of paratuberculosis in northern Italy dairy goat farms was confirmed. The welfare and biosecurity assessment protocol proved to be an accurate tool, capable of identifying critical points for managing health, welfare and productivity.

Ante-mortem and post-mortem inspection at slaughter are performed by Official Veterinarians and it is essential to identify alterations/lesions, which can make organs/carcasses unsuitable for human consumption. Obviously, carcass condemnation must be regarded as a highly undesirable event for the entire swine industry chain, as it represents the total failure of a long period of challenging work. Therefore, it seems valuable to estimate the prevalence and causes of carcasses condemnation, in order to preserve consumers’ health and pig farming profitability. Bearing that in mind, the present study aimed at providing a reliable picture of the condemnation of porcine carcasses in Italy, with a special emphasis being placed upon pathological findings. Data were collected in a high-throughput abattoir located in northern Italy, where a total of 2,062,278 pigs were slaughtered during the period of study (2021–2022). Overall, 1362 pigs were considered unfit for slaughtering after ante-mortem inspection, mostly because of death during transportation or in resting pens, after their arrival at the abattoir. Moreover, 2007 carcasses were condemned after post-mortem inspection. The most common causes of condemnation were severe and diffuse peritonitis, disseminated abscesses, jaundice, and erysipelas. In our opinion, the present survey may provide useful and updated information about the condemnation of pig carcasses in Italy. At the same time, it highlights the need to collect data in a more systematic and standardized way, thus making possible their comparison over time and among different geographic areas.

In recent years, the growth of wild ungulates has increased the focus on their health monitoring. In particular, the health status of wild boars is relevant for the economic impact on the pig industry. The Emilia-Romagna region activated a wildlife monitoring plan to better evaluate the health status of the wild boar population. Between 2011 and 2021, samples of found dead and hunted wild boar have been examined for trichinellosis, tuberculosis, brucellosis, african swine fever, classical swine fever, Aujeszky’s disease, swine vesicular disease, and swine influenza A. Trichinella britovi was identified in 0.001% of the examined wild boars; neither M. bovis nor M. tuberculosis were found in M. tuberculosis complex positive samples; 2.3% were positive for Brucella suis; 29.4% of the sera were positive for Aujeszky’s disease virus; and 0.9% of the samples were positive for swine influenza A virus. With an uncertain population estimate, the number of animals tested, the number of positives, and the sampling method do not allow us to make many inferences but suggest the need to implement and strengthen the existing surveillance activity, as it seems to be the only viable alternative for safeguarding animal and human health.

Streptococcus agalactiae (group B Streptococcus, GBS) is a causative agent of mastitis in dairy cattle, mainly causing a subclinical disease associated with a high somatic cell count (SCC), and a consequent decrease in production yield and quality of milk. GBS has been almost eradicated in many Northern European countries, but there are warnings of its re-emergence as a zoonotic threat. In Italy, only two regions carry out a GBS control program: Lombardy and Emilia-Romagna. In Emilia-Romagna, the program has been in place since 2019 and provides for the bacteriological culture of bulk-tank milk (BTM) of all dairy farms every 6 months and the voluntary application of herd eradication programs in the case of positive results. To assess the progress of the program in Emilia Romagna, in terms of herd-level prevalence and GBS transmission between herds, we analyzed the results of 17,056 BTM cultures from 2,831 dairy herds, sampled bi-annually in the period 2019–2021 (six rounds total). The impact of GBS infection on SCC and milk production was also evaluated. The results show a decreasing trend in both the incidence rate (from 3.0 to 1.5%) and apparent prevalence (from 8.9 to 5.2%) of GBS over the study period. By using a susceptible-infected-susceptible (SIS) model for the estimation of the transmission parameters, a basic reproductive number R0 of 1.4 was calculated, indicating an active spread of GBS in the dairy cattle population of the Emilia-Romagna region. GBS infected farms have a consistently higher BTM SCC than negative ones (+77,000 cells/ml), corresponding to a 0.4 kg/cow/day milk loss. Moreover, GBS infected herds resulted in almost three times more likelihood of having non-marketable milk by exceeding the legal SCC limit. This study demonstrates the need to maintain the current control program against GBS to lower its occurrence and prevent significant market losses to farmers.

American Foulbrood (AFB) is a contagious and severe brood disease of honey bees caused by the spore-forming bacterium Paenibacillus larvae. The identification of honey bee colonies infected by P. larvae is crucial for the effective control of AFB. We studied the possibility of identifying the infection levels by P. larvae in honey bee colonies through the examination of powdered sugar samples collected in the hives. The powdered sugar was dusted on the top bars of honeycombs and collected from a sheet paper placed at the bottom of the hive. Three groups of honey bee colonies were examined: Group A1- colonies with clinical symptoms of AFB (n = 11); Group A2 – asymptomatic colonies located in apiaries with colonies showing symptoms of AFB (n = 59); Group B – asymptomatic colonies located in apiaries without cases of the disease (n = 49). The results showed that there was a significant difference in spore counting between Groups and that the spore load in sugar samples was always consistent with the clinical conditions of the colonies and with their belonging to AFB-affected apiaries or not. Based on the obtained results the cultural examination of powdered sugar samples collected from hives could be an effective tool for the quantitative non-destructive assessment of P. larvae infections in honey bee colonies.

In the last decade, an upsurge of human leishmaniasis has been reported in the Emilia-Romagna region, Northeast Italy. Epidemiologic data have raised doubts about the role of dogs as the main reservoirs for Leishmania infantum. In the present study, a total of 1077 wild animals were screened for L. infantum DNA in earlobe and spleen samples from 2019 to 2022. The lymph nodes were tested only in 23 animals already positive in the earlobe and/or spleen. A total of 71 (6.6%) animals resulted positive in at least one of the sampled tissues, including 3/18 (16.7%) wolves, 6/39 (15.4%) European hares, 38/309 (12.3%) roe deer, 1/11 (9.1%) red deer, 8/146 (4.9%) wild boars, 13/319 (4.1%) red foxes, 1/54 (1.9%) porcupine, and 1/59 (1.7%) European badger. Most of the infected animals (62/71) tested positive only in the earlobe tissue, only four animals (two roe deer and two wild boars) tested positive only in the spleen, and five animals (three roe deer and two red foxes) resulted positive for both tissues. L. infantum DNA was detected in the lymph nodes of 6/23 animals. L. infantum detection occurred in all seasons associated with low real-time PCR Ct values. Further research is needed in order to clarify the role of wildlife in the re-emerging focus of leishmaniasis in Northeast Italy.

Paenibacillus larvae (P. larvae) is responsible for American foulbrood (AFB), the most severe bacterial disease of honeybees. The enumeration of P. larvae spores in substrates taken from hives allows for the identification of the contamination levels of the colonies, mostly in those with atypical symptoms or with asymptomatic infections; in these cases, it is essential for the effective control of American foulbrood (AFB). In this work we described a new quantitative TaqMan® probe-based real-time PCR (qPCR) assay, targeting the 16S rRNA gene of P. larvae, used for the quantification of P. larvae spores in powdered sugar samples collected from hives, in comparison to the culture. A total of 105 colonies were selected, belonging to 10 apiaries with different levels of infection, located in northern Italy. The proportions of positive colonies was 54% (57/105) with the culture and 66% (69/105) with qPCR. A significant difference between the two methods was found with McNemar’s test (p = 0.02). Out of the 51 positive samples by both methods, 45 showed higher infection by qPCR compared to the culture. A close concordance with the clinical–epidemiological status of the hives was observed by both methods, with higher infection levels found by qPCR.

With several human cases reported annually since 2008 and the unapparent risk of infection of blood donors, the West Nile virus (WNV) is emerging as an important health issue in Europe. Italy, as well as other European countries, experienced a recrudescence of the virus circulation in 2018, which led to an increased number of human cases. An integrated surveillance plan was activated in the Emilia-Romagna and Lombardy regions (Northern Italy) since 2008 in order to monitor the intensity and timing of WNV circulation. A fundamental part of this plan consists in entomological surveillance. In 2018, the surveillance plan made it possible to collect 385,293 mosquitoes in 163 stations in the two Regions. In total 269,147 Culex mosquitoes were grouped into 2,337 pools and tested for WNV, which was detected in 232 pools. Circulation started in the central part of the Emilia-Romagna region in the middle of June, about one month before the previous seasons. Circulation suddenly expanded to the rest of the region and reached the Lombardy region in the middle of July. WNV circulated more intensively in the eastern part of the surveyed area, as confirmed by the highest number of human cases. A relationship between the number of mosquitoes collected and the virus incidence emerged, but the data obtained highlighted that the probability of detecting the virus in a given site was less than expected with a higher number of collected mosquitoes. A significant relationship was observed between the temperature recorded one week before the sampling and the number of collected mosquitoes, as well as between the estimated number of WNV-positive mosquitoes and the temperature recorded two weeks before the sampling. The two weeks delay in the influence of temperature on the positive mosquitoes is in line with the time of the virus extrinsic incubation in the mosquito. This finding confirms that temperature is one of the principal drivers in WNV mosquito infection. The surveillance system demonstrated the ability to detect the virus circulation early, particularly in areas where circulation was more intense. This allowed evaluating the effect of mosquito abundance and weather factors on virus circulation.

West Nile (WNV) and Usutu (USUV) viruses are mosquito-borne flaviviruses. Thanks to their importance as zoonotic diseases, a regional plan for surveillance of Arboviruses was implemented in Emilia-Romagna in 2009. The province of Ferrara belongs to the Emilia-Romagna region, and it is an endemic territory for these viruses, with favorable ecological conditions for abundance of mosquitoes and wild birds. From 2015 to 2019, we collected 1842 dead-found birds at a wildlife rehabilitation center, which were analysed by three different PCRs for the detection of WNV and USUV genomes. August was characterized by the highest infection rate for both viruses. Columbiformes scored the highest USUV prevalence (8%), while Galliformes and Strigiformes reported the highest prevalence for WNV (13%). Among Passeriformes (the most populated Order), Turdus merula was the most abundant species and scored the highest prevalence for both viruses. To optimize passive surveillance plans, monitoring should be focused on the summer and towards the avian species more prone to infection by both viruses.

Since 2013 in Emilia-Romagna, Italy, surveillance information generated in the public health and in the animal health sectors has been shared and used to guide public health interventions to mitigate the risk of West Nile virus (WNV) transmission via blood transfusion. The objective of the current study was to identify and estimate the costs and benefits associated with this One Health surveillance approach, and to compare it to an approach that does not integrate animal health information in blood donations safety policy (uni-sectoral scenario). Costs of human, animal, and entomological surveillance, sharing of information, and triggered interventions were estimated. Benefits were quantified as the averted costs of potential human cases of WNV neuroinvasive disease associated to infected blood transfusion. In the 2009-2015 period, the One Health approach was estimated to represent a cost saving of €160,921 compared to the uni-sectoral scenario. Blood donation screening was the main cost for both scenarios. The One Health approach further allowed savings of €1.21 million in terms of avoided tests on blood units. Benefits of the One Health approach due to short-term costs of hospitalization and compensation for transfusion-associated disease potentially avoided, were estimated to range from €0 to €2.98 million according to the probability of developing WNV neuroinvasive disease after receiving an infected blood transfusion.

Chemical treatments are commonly adopted for poultry house sanitation. In fact, ordinary floor disinfection is needed to deplete the pathogenic population (i.e. various species of bacteria and fungi) and reduce the risk of meat contamination. The increasing focus on the health of consumers and operators, as well as on food quality, has led farmers to consider alternative environmentally friendly methods. Research was carried out to set up a new machine for floor disinfection of poultry houses by open flame. The trials were run in controlled conditions in the laboratory of the University of Pisa, Italy, and on a private farm. The first experiment consisted of a series of test bench trials carried out to evaluate the efficacy and the adjustment of liquefied petroleum gas (LPG)-fed open flame burners on pre-inoculated steel plates. In the second experiment, the operative parameters of a custom-built 1.5 m wide mounted flaming machine were determined and the biological effects of the treatment were compared to ordinary chemical treatments. The results obtained were very promising. Test bench trials showed a 4-log reduction in E. coli, and microbial determinations carried out on-farm did not show any difference between thermal and chemical treatment. In addition, the cost estimation showed that thermal disinfection is approximately 4-fold cheaper than chemical sanitation methods. The effective working capacity of the machine was approximately 1700 m2 h–1, and the LPG consumption was approximately 16 kg per 1000 m2. Flame disinfection of poultry grow-out facilities could represent a valid alternative to chemical disinfection.

In 2008, after the first West Nile virus (WNV) detection in the Emilia-Romagna region, a surveillance system, including mosquito- and bird-based surveillance, was established to evaluate the virus presence. Surveillance was improved in following years by extending the monitoring to larger areas and increasing the numbers of mosquitoes and birds tested.A network of mosquito traps, evenly distributed and regularly activated, was set up within the surveyed area. A total of 438,558 mosquitoes, grouped in 3,111 pools and 1,276 birds (1,130 actively sampled and 146 from passive surveillance), were tested by biomolecular analysis. The survey detected WNV in 3 Culex pipiens pools while Usutu virus (USUV) was found in 89 Cx. pipiens pools and in 2 Aedes albopictus pools. Two birds were WNV-positive and 12 were USUV-positive. Furthermore, 30 human cases of acute meningoencephalitis, possibly caused by WNV or USUV, were evaluated for both viruses and 1,053 blood bags were tested for WNV, without any positive result.Despite not finding symptomatic human WNV infections during 2010, the persistence of the virus, probably due to overwintering, was confirmed through viral circulation in mosquitoes and birds, as well as for USUV. In 2010, circulation of the two viruses was lower and more delayed than in 2009, but this decrease was not explained by the relative abundance of Cx. pipiens mosquito, which was greater in 2010. The USUV detection in mosquito species confirms the role of Cx. pipiens as the main vector and the possible involvement of Ae. albopictus in the virus cycle. The effects of meteorological conditions on the presence of USUV-positive mosquito pools were considered finding an association with drought conditions and a wide temperature range. The output produced by the surveillance system demonstrated its usefulness and reliability in terms of planning public health policies.

BACKGROUND: In recent years human diseases due to mosquito-borne viruses were increasingly reported in Emilia-Romagna region (Italy), from the chikungunya virus in 2007 to the West Nile virus (WNV) in 2008. An extensive entomological survey was performed in 2009 to establish the presence and distribution of mosquito arboviruses in this region, with particular reference to flaviviruses. METHODOLOGY/PRINCIPAL FINDINGS: From May 6 to October 31, a total of 190,516 mosquitoes were sampled in georeferenced stations, grouped in 1,789 pools according date of collection, location, and species, and analyzed by reverse transcription polymerase chain reaction (RT-PCR) to detect the presence of RNA belong to Flavivirus genus. WNV was detected in 27 mosquito pools, producing sequences similar to those of birds and human strains obtained in 2008 outbreak, pointed out the probable virus overwintering. Isolation of WNV was achieved from one of these pools. Moreover 56 pools of mosquitoes tested positive for Usutu virus (USUV). Most PCR positive pools consisted of Culex pipiens, which also was the most analyzed mosquito species (81.4% of specimens); interestingly, USUV RNA was also found in two Aedes albopictus mosquito pools. Simultaneous circulation of WNV and USUV in the survey area was highlighted by occurrence of 8 mosquito WNV- and USUV-positive pools and by the overlaying of the viruses "hot spots", obtained by kernel density estimation (KDE) analysis. Land use of sampled stations pointed out a higher proportion of WNV-positive Cx. pipiens pool in rural environments respect the provenience of total sampled pool, while the USUV-positive pools were uniformly captured in the different environments. CONCLUSIONS/SIGNIFICANCE: Obtained data highlighting the possible role of Cx. pipiens mosquito as the main vector for WNV and USUV in Northern Italy, and the possible involvement of Ae. albopictus mosquito in USUV cycle. The described mosquito-based surveillance could constitute the foundation for a public health alert system targeting mosquito borne arboviruses.