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Tick-borne encephalitis was limited to northeast portions of Italy. We report in Lombardy, a populous region in the northwest, a chamois displaying clinical signs of tickborne encephalitis virus that had multiple virus-positive ticks attached, as well as a symptomatic man. Further, we show serologic evidence of viral circulation in the area.

Bovine viral diarrhea (BVD) is a major cause of economic losses in the global cattle industry, particularly in countries characterized by intensive livestock production systems. Pestivirus tauri, formerly known as Bovine viral diarrhea virus type 2 (BVDV-2), is the current taxonomic designation according to the International Committee on Taxonomy of Viruses (ICTV). Between 2005 and 2018, Pestivirus tauri was detected in cattle herds in mainland Italy, particularly in the Lombardy region. Four viral strains were successfully isolated in cell cultures and subjected to whole-genome sequencing. Phylogenetic reconstruction placed all Italian isolates within the Pestivirus tauri subgenotype c, a lineage encompassing strains reported in Asia, Europe and the United States. Consistently, comparative sequence identity analyses indicated the highest similarity with the Parker strain (USA, 1991) and the Potsdam 1600 strain (Germany, 2000). These results contribute to a more detailed understanding of Pestivirus tauri genomic architecture and evolutionary dynamics, providing a valuable resource for comparative genomic studies. Such data are crucial for exploring viral diversity and evolution, optimizing the design of diagnostic primers and probes, and advancing insights into the molecular epidemiology of Pestivirus.

Bioaugmentation, defined as the strategic incorporation of specifically selected microbial biomass into contaminated environments, can significantly enhance the biodegradation of pollutants and is extensively employed in soil bioremediation efforts. A multistep screening process was applied to develop an autochthonous microbial consortium, including (i) hydrocarbonoclastic strain isolation from soil chronically contaminated with petroleum hydrocarbons, (ii) bacterial selection according to genomic and functional traits, and (iii) consortium validation in the native contaminated soil through microcosm experiments. The selection of strains with the ability to degrade alkanes and aromatic hydrocarbons on synthetic media was further supported by genomic analysis, delivering a consortium with complementary degradative properties. The outcomes of the microcosm experiments corroborated the efficacy of the selected indigenous consortium, demonstrating that the combination of Acinetobacter guillouiae, A. radioresistens, and Pseudomonas zarinae as an inoculum in the bioaugmentation strategy was successful in achieving the removal of up to 26% and 76% of linear and polycyclic aromatic hydrocarbons, respectively, thereby effectively addressing areas where natural attenuation was insufficient.

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.

The correct identification of mosquito vectors is often hampered by the presence of morphologically indiscernible sibling species. The Maculipennis complex is one of these groups that include both malaria vectors of primary importance and species of low/negligible epidemiological relevance, of which distribution data in Italy are outdated. Our study was aimed at providing an updated distribution of Maculipennis complex in Northern Italy through the sampling and morphological/molecular identification of specimens from five regions. The most abundant species was Anopheles messeae (2032), followed by Anopheles maculipennis s.s. (418), Anopheles atroparvus (28) and Anopheles melanoon (13). Taking advantage of ITS2 barcoding, we were able to finely characterize tested mosquitoes, classifying all the Anopheles messeae specimens as Anopheles daciae , a taxon with debated rank to which we referred as species inquirenda ( sp. inq. ). The distribution of species was characterized by Ecological Niche Models (ENMs), fed by recorded points of presence. ENMs provided clues on the ecological preferences of the detected species, with An. daciae sp. inq. linked to stable breeding sites and An. maculipennis s.s. more associated to ephemeral breeding sites. We demonstrate that historical Anopheles malaria vectors are still present in Northern Italy.

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

Hepatitis E virus (HEV) causes viral hepatitis, posing a risk to vulnerable populations. Although swine are established reservoirs, the involvement of other animal species remains undefined. This study investigated 301 livers from naturally deceased European hedgehogs collected from wildlife centres (northern Italy), to determine their role in HEV transmission, spread and epidemiology, considering their proximity to urban areas. No positive samples were found by Real-time PCR, suggesting that hedgehogs play a marginal role in HEV epidemiology. Further studies are required to investigate other potential wild reservoirs to gain insight into HEV transmission dynamics, given the global impact of HEV-related diseases.

Coronaviruses (CoVs) are widespread and highly diversified in wildlife and domestic mammals and can emerge as zoonotic or epizootic pathogens and consequently host shift from these reservoirs, highlighting the importance of veterinary surveillance. All genera can be found in mammals, with α and β showing the highest frequency and diversification. The aims of this study were to review the literature for features of CoV surveillance in animals, to test widely used molecular protocols, and to identify the most effective one in terms of spectrum and sensitivity. We combined a literature review with analyses in silico and in vitro using viral strains and archive field samples. We found that most protocols defined as pan-coronavirus are strongly biased towards α- and β-CoVs and show medium-low sensitivity. The best results were observed using our new protocol, showing LoD 100 PFU/mL for SARS-CoV-2, 50 TCID50/mL for CaCoV, 0.39 TCID50/mL for BoCoV, and 9 ± 1 log2 ×10−5 HA for IBV. The protocol successfully confirmed the positivity for a broad range of CoVs in 30/30 field samples. Our study points out that pan-CoV surveillance in mammals could be strongly improved in sensitivity and spectrum and propose the application of a new RT-PCR assay, which is able to detect CoVs from all four genera, with an optimal sensitivity for α-, β-, and γ-.

Phleboviruses transmitted by phlebotomine sandflies are endemic in the Mediterranean basin. Toscana phlebovirus (TOSV), Sicilian phlebovirus (SFSV), and Naples phlebovirus (SFNV) are responsible of summer fever, with well-known pathogenic potential for humans ranging from asymptomatic to mild fever, in addition to neuro-invasive infections during summer. Although TOSV, in particular, is a significant and well-known human pathogen, SFVs remain neglected, with many gaps in the relevant knowledge. Sero-epidemiological studies and case reports recently showed a geographical wider distribution than previously considered, although the real incidence of phleboviruses infections in the Mediterranean area is still unknown. Here we retrospectively evaluated the circulation of phleboviruses during summer seasons between 2007 and 2019 in 649 patients showing neurological symptoms using both molecular and serological approaches. We found that 42/649 (6.5%) subjects experienced phlebovirus infection and only 10/42 cases were detected by molecular assays, whereas the other 32/42 were identified using serological approaches, including neutralization assays. During the 2013 summer, an outbreak in the Lombardy region is described because the prevalence of phlebovirus infection reached 37.2% (19/51 subjects). Interestingly, only 5/19 (26.5%) reported traveling in endemic areas. Of note, no cross-neutralization was observed between different strains tested, showing the possibility to be reinfected by newly discovered phlebovirus strains. In conclusion, phlebovirus infections are still inadequately considered by physicians and are generally underestimated. However, based on our results, sandfly fever viruses should be routinely included in diagnostic panels during summer period, including in Northern Italy.

We isolated Issyk-Kul virus (ISKV) from a bat sampled from Italy in 2021 and conducted ISKV-specific surveillance in bats collected in Italy during 2017-2023. ISKV circulation among synanthropic and sedentary species of bat, such as Savi's pipistrelle bat (Hypsugo savii) in northern Italy, may have public health implications in this region.