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West Nile disease is a vector-borne disease caused by West Nile virus (WNV), involving mosquitoes as vectors and birds as maintenance hosts. Humans and other mammals can be infected via mosquito bites, developing symptoms ranging from mild fever to severe neurological infection. Due to the worldwide spread of WNV, human infection risk is high in several countries. Nevertheless, there are still several knowledge gaps regarding WNV dynamics. Several aspects of transmission taking place between birds and mosquitoes, such as the length of the infectious period in birds or mosquito biting rates, are still not fully understood, and precise quantitative estimates are still lacking for the European species involved. This lack of knowledge affects the precision of parameter values when modelling the infection, consequently resulting in a potential impairment of the reliability of model simulations and predictions and in a lack of the overall understanding of WNV spread. Further investigations are thus needed to better understand these aspects, but field studies, especially those involving several wild species, such as in the case of WNV, can be challenging. Thus, it becomes crucial to identify which transmission processes most influence the dynamics of WNV. In the present work, we propose a sensitivity analysis to investigate which of the selected epidemiological parameters of WNV have the largest impact on the spread of the infection. Based on a mathematical model simulating WNV spread into the Lombardy region (northern Italy), the basic reproduction number of the infection was estimated and used to quantify infection spread into mosquitoes and birds. Then, we quantified how variations in four epidemiological parameters representing the duration of the infectious period in birds, the mosquito biting rate on birds, and the competence and susceptibility to infection of different bird species might affect WNV transmission. Our study highlights that knowledge gaps in WNV epidemiology affect the precision in several parameters. Although all investigated parameters affected the spread of WNV and the modelling precision, the duration of the infectious period in birds and mosquito biting rate are the most impactful, pointing out the need of focusing future studies on a better estimate of these parameters at first. In addition, our study suggests that a WNV outbreak is very likely to occur in all areas with suitable temperatures, highlighting the wide area where WNV represents a serious risk for public health.

Background West Nile virus (WNV) is an emerging vector-borne pathogen that is becoming increasingly prevalent in temperate regions. The development of effective intervention strategies is crucial for limiting its spread; however, the adaptability and ubiquity of mosquitoes, combined with the complexity of the WNV transmission cycle, continue to hinder its eradication. Methods This study employs a deterministic compartmental model to evaluate the effectiveness of ten intervention strategies targeting either the mosquito (vector) or avian (host) population in the Lombardy region of Italy. Results Vector-targeted interventions were more effective than host-targeted measures, with breeding site reduction and larvicide treatments demonstrating the greatest efficacy. In contrast, interventions targeting adult mosquitoes, including adulticide treatments and elimination of overwintering mosquitoes, showed moderate efficacy. Furthermore, the impact of eliminating overwintering mosquitoes gradually diminished over time. Host-targeted strategies, such as bird population reduction, were ineffective and, in some cases, led to increased WNV transmission. The efficacy of all interventions varied temporally, peaking in mid-summer. Conclusions These findings highlight the importance of prioritising mosquito control, particularly targeting immature stages, to mitigate WNV outbreaks. Our study highlights the critical role of mathematical modelling in designing effective intervention strategies. By providing a structured framework to evaluate and predict the outcomes of various approaches, modelling can aid disease control while optimising resource allocation and minimising environmental impact. Mathematical models, therefore, prove to be powerful tools for balancing public health goals with sustainable practices. Graphical Abstract

In the last two years, there have been three major epidemic seasons in the territory of the European Union and the HPAI epizootic in 2021–2022 is the most severe in recent history. In Italy, the disease was introduced to dense poultry areas with serious economic consequences for the entire sector. In Lombardy, the analysis of the risk factors was carried out, also taking into account the density of domestic birds. In the most affected areas, 66.7% of the outbreaks occurred in the areas with the highest poultry density and the likelihood of an outbreak occurring increased with an increase in the density of birds per km2. In cells 10 × 10 km with a density greater than 10,000 birds/km2, the probability of outbreak occurrence was over 66.7%. The provinces involved in the last epidemic were the same involved in previous epidemics and, given the risk factors present in the area, it is plausible that the risk remains high also for future epidemic seasons. Therefore, to avoid the repetition of similar events, certain control measures shall be strengthened and vaccination considered as a complementary tool for the control of HPAI virus in risk areas.

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 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.

This study investigated the species composition and density of sand flies in the Lombardy region (Northern Italy). Sand flies were collected using CDC traps baited with CO2 (CO2–CDC traps) between June and August 2021. A total of 670 sand flies were collected. The specimens were identified as seven species belonging to two genera, Phlebotomus and Sergentomyia, namely, S. minuta, Ph. perniciosus, Ph. perfiliewii, Ph. neglectus, Ph. mascitti, Ph. papatasi, and Ph. ariasi. Phlebotomus perniciosus was the most abundant species (87.76%), followed by Ph. perfiliewii (7.31%), Ph. neglectus (3.13%), S. minuta (0.75%), Ph. mascitti (0.6%), Ph. papatasi (0.3%), and Ph. ariasi, for which only one specimen was identified. Among these identified species, five are considered vectors of Leishmania, which causes cutaneous and visceral leishmaniasis. As vector presence increases the risk of vector-borne leishmaniasis, these results suggest that Northern Italy could be a potential area of pathogen circulation over the next few years. These preliminary results suggest that the risk of borne leishmaniasis is high in this region of Northern Italy. Monitoring the distribution of sand fly species in areas suitable for their persistence is important for control programs aimed at reducing the risk of leishmaniasis infection.

The western honey bee (Apis mellifera) represents a key pollinator for both crops and wild plants, and its global decline raises serious concerns for ecosystem stability and agricultural productivity. Several biotic and abiotic factors are responsible for colony losses, including alterations in the bee microbiota, which is essential for host metabolism, development, and immune responses. In this study, we employed both molecular protocols and metagenomic approaches based on Next-Generation Sequencing (NGS) to characterize the microbial composition and identify commensal, symbiotic, and pathogenic microorganisms, both known and emerging, associated with A. mellifera colonies from 20 apiaries across the Italian territory. Molecular screening revealed Vairimorpha ceranae, Lotmaria passim, Crithidia mellificae and several viruses, including Sacbrood virus (SBV), Black Queen Cell virus (BQCV), Deformed Wing virus (DWV), Chronic Bee Paralysis virus (CBPV) and Acute Bee Paralysis virus (ABPV). 16S rRNA gene sequencing highlighted a bacterial community mainly composed of the Lactobacillus, Gilliamella, and Snodgrassella genera. Virome analysis detected members belonging to the families Dicistroviridae and Iflaviridae, as well as previously unreported viruses in Italy, such as Apis rhabdovirus (ARV-1, ARV-2), Bee Macula-like virus (BeeMLV), and Lake Sinai virus (LSV). This research expands current knowledge of the A. mellifera metagenome, offering valuable insights for epidemiological surveillance and diagnostic assay development.

Porcine Respirovirus 1 (PRV1) is an enveloped, single-stranded, negative-sense RNA virus belonging to the genus Respirovirus within the Paramyxoviridae family. Since its first detection in China in 2013, PRV1 has been identified in several American and European countries. Although its pathogenicity is uncertain, recent studies have suggested that it may play a role in the Porcine Respiratory Disease Complex (PRDC) because of its capacity to replicate in the upper and lower respiratory tracts. This study aimed to determine the spread of PRV1 in Northern Italy and the phylogeny of the isolates. Therefore, PRV1 was investigated using real-time RT-PCR in 902 samples collected from September 2022 to September 2023 from pigs with respiratory symptoms in North Italy. Fourteen (1.55%) samples tested as PRV1-positive. The full-length fusion (F) gene, which codifies for a major surface protein, was amplified and used for phylogenetic analysis to help carry out molecular epidemiological studies on this virus. In addition, swine influenza virus (SIV) and porcine reproductive and respiratory syndrome virus (PRRSV) infections were detected in most of the PRV1-positive samples. In conclusion, we report the detection of PRV1 in Italy and discuss its potential role as a co-factor in causing the Porcine Respiratory Disease Complex.

The first events of bee decline in Italy were reported during 1999. Since then, population decline has frequently been reported in Lombardy. In this study, the association between bee decline and the type of land surrounding the apiary was evaluated. A risk map was developed to identify areas with the highest risk of decline. Apiaries in Lombardy were selected from the national beekeeping database (BDA). The study period was from 2014 to 2016. Apiaries were deemed “declined” if they reported at least one event of decline or tested positive for plant protection products; apiaries were “not declined” if they did not report any events of bee decline during the study period. Out of 14,188 apiaries extracted from the BDA, 80 were considered declined. The probability of an apiary being declined increases by 10% in orchards and by 2% in arable land for each additional km2 of land occupied by these crops. The study showed an association between bee decline and the type of territory surrounding the apiaries, and the areas at the greatest risk of decline in Lombardy were identified. This information can be used by Veterinary Services as a predictive parameter for planning prevention and control activities.

In Italy, the West Nile Virus surveillance plan considers a multidisciplinary approach to identify the presence of the virus in the environment (entomological, ornithological, and equine surveillance) and to determine the risk of infections through potentially infected donors (blood and organ donors). The costs associated with the surveillance program for the Lombardy Region between 2014 and 2018 were estimated. The costs of the program were compared with a scenario in which the program was not implemented, requiring individual blood donation nucleic acid amplification tests (NAT) to detect the presence of WNV in human samples throughout the seasonal period of vector presence. Considering the five-year period, the application of the environmental/veterinary surveillance program allowed a reduction in costs incurred in the Lombardy Region of 7.7 million EUR. An integrated surveillance system, including birds, mosquito vectors, and dead-end hosts such as horses and humans, can prevent viral transmission to the human population, as well as anticipate the detection of WNV using NAT in blood and organ donors. The surveillance program within a One Health context has given the possibility to both document the expansion of the endemic area of WNV in northern Italy and avoid most of the NAT-related costs.

West Nile disease is a vector-borne disease caused by West Nile virus (WNV), involving mosquitoes as vectors and birds as maintenance hosts. Humans and other mammals can be infected via mosquito bites, developing symptoms ranging from mild fever to severe neurological infection. Due to the worldwide spread of WNV, human infection risk is high in several countries. Nevertheless, there are still several knowledge gaps regarding WNV dynamics. Several aspects of transmission taking place between birds and mosquitoes, such as the length of the infectious period in birds or mosquito biting rates, are still not fully understood, and precise quantitative estimates are still lacking for the European species involved. This lack of knowledge affects the precision of parameter values when modelling the infection, consequently resulting in a potential impairment of the reliability of model simulations and predictions. Further investigations are thus needed to better understand these aspects, but field studies, especially those involving several wild species, such as in the case of WNV, can be challenging. Thus, it becomes crucial to identify which mechanisms most influence the dynamics of WNV transmission. In the present work, we propose a sensitivity analysis to identify the parameters that have the largest impact on the spread of the infection. Based on a mathematical model simulating WNV spread into the Lombardy region (northern Italy), the basic reproduction number of the infection was estimated and used to quantify infection spread into mosquitoes and birds. Then, we quantified how variations in the duration of the infectious period in birds, the mosquito biting rate on birds, and the competence and susceptibility to infection of different bird species might affect WNV transmission. Our study highlights that all considered parameters can affect the spread of WNV, but the duration of the infectious period in birds and mosquito biting rate are the most impactful, highlighting the need for further studies to better estimate these parameters. In addition, our study suggests that a WNV outbreak is very likely to occur in all areas with suitable temperatures, highlighting the wide area where WNV represents a serious risk for public health. Competing Interest Statement The authors have declared no competing interest.