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Usutu virus (USUV) is a mosquito-borne virus belonging to the family Flaviviridae, genus Flavivirus. Natural transmission cycle of USUV involves mosquitoes and birds, so humans and other mammals are considered incidental hosts. In this study, USUV infection was diagnosed in all wild blackbirds, collected from July to September 2018 in a wildlife recovery center in the province of Bologna, in the Emilia-Romagna region, northern Italy. All blackbirds showed neurological clinical signs, such as overturning, pedaling, and incoordination. Moreover, the subjects died shortly after arriving at the hospitalization center. Virological investigations were performed by real-time PCR on frozen samples of the spleen, kidney, myocardium, and brain for the detection of Usutu (USUV) and West Nile (WNV) viruses. The small and large intestine were used as a matrix for the detection of Newcastle disease virus (NDV). All 56 subjects with neurological clinical signs were positive for USUV, only one subject (1.8%) tested positive for WNV, and no subject was positive for NDV. The most represented age class was class 1 J (58.9%), followed by class 3 (25.0%), and lastly from class 4 (16.1%). Most of the blackbirds before dying were in good (51.8%) and fair (39.3%) nutritional status, while only five subjects (8.9%) were cachectic. The USUV genomes detected in the blackbirds of this study fall within the sub-clade already called EU2 that has been detected since 2009 in the Emilia-Romagna region. Neurological clinical signs in USUV-affected blackbirds are still widely discussed and there are few works in the literature. Although our results require further studies, we believe them to be useful for understanding the clinical signs of Usutu virus in blackbirds, helping to increase the knowledge of this zoonotic agent in wild species and to understand its effect on the ecosystem. The goal of this study was to report—in the context of the regional passive surveillance program—the detection of USUV RNA in its most important amplifying host, the common blackbird, when showing clinical signs before death. [ABSTRACT FROM AUTHOR]

During influenza surveillance activities carried out from 1999 to 2001 in wild ducks, 311 cloacal swabs were collected in Central Italy. A total of 20 avian influenza viruses (AIVs) were isolated, that belonged to eight different subtypes. Five of these subtypes had never been isolated before in the area under study, including two H7N3 viruses obtained from mallards in Autumn 2001. One year after, outbreaks of AI caused by H7N3 LP viruses occurred and rapidly spread among commercial poultry farms in Northern Italy. We compared the H7N3 duck strains with the first two H7N3 turkey isolate, and a high level of similarity between the two viruses was found in all genes, except for the NA. The N3 genes of domestic poultry strains showed a 23-amino acid deletion in the stalk region, indicating an initial adaptation of wild waterfowl viruses to terrestrial bird species. Phylogenetic data are also discussed. This is the first report in which a domestic poultry AIV and its direct wild bird counterpart are described. [Copyright &y& Elsevier]

We report the results of a 6-year serological and virological monitoring performed in ducks and coots in Italy, in order to assess the degree of influenza A virus circulation in these birds during wintering. A total of 1039 sera collected from 1992 to 1998 was screened by a double antibody sandwich blocking ELISA (NP-ELISA): seroprevalence of antibodies to influenza A viruses was significantly higher in ducks compared to coots (52.2% vs. 7.1%, respectively). The hemagglutination-inhibition (HI) assay, performed on NP-ELISA positive sera, showed that 16.9% of these duck sera and 33.3% of these coot sera had antibodies to at least one influenza virus HA subtype: ducks showed HI antibodies against most of the HA subtypes, except for the H3, H4, H7, and H12; coots were seropositive to the H3 and H10 subtypes, only. From 1993 to 1998, 22 virus strains were obtained from 802 cloacal swabs, with an overall virus isolation frequency of 2.7%. Viruses belonging to the H1N1 subtype were by far the most commonly circulating strains (18/22) and were isolated mainly from ducks (17/18). The remaining viruses were representative of the H10N8, H5N2 and H3N8 subtypes. Our data indicate some differences between influenza A virus circulation in sympatric ducks and coots and a significant antigenic diversity between some reference strains and viruses recently isolated in Italy. [Copyright &y& Elsevier]

Although many studies highlighted the potential of COVID-19 to reshape existing models of wildlife management, empirical research on this topic has been scarce, particularly in Europe. We investigated the potential of COVID-19 pandemic to accelerate the ongoing decline in an aging population of recreational hunters in Italy. Namely, we modelled spatiotemporal trends between 2011 and 2021 in the number of recreational hunters in 50 Italian provinces with a varying incidence of COVID-19, and temporally delayed waves of infection. Compared to projections from 2011-2019 data, we detected a lower number of hunters who enrolled for the hunting season, both in 2020 (14 provinces) and in 2021 (15 provinces). The provinces with the highest incidence of COVID-19 in the Lombardy and Emilia-Romagna regions were also those experiencing the most marked decrease in hunting participation. Our findings revealed that a wildlife management system based on recreational hunting can be rapidly destabilized by epidemics and their associated public health measures, particularly when the average age of hunters is high, like in Italy. Considered the high incidence attained by COVID-19 in many European countries, where hunters are pivotal for the management of large ungulates and where they were already declining before the pandemic, our findings call for further large-scale research about the impact of COVID-19 on hunting participation., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Cerri et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Background: The wild boar (Sus scrofa) and the Apennine wolf (Canis lupus italicus) are two wild species that have both increased their presence in the Italian territory, albeit in varying numbers. They can be occasionally found in peri-urban areas as well. Both of these species can serve as intermediate hosts for Toxoplasma gondii, as they can become infected either through the consumption of oocysts found in water, soil, or on vegetables, or through the ingestion of meat containing bradyzoites. Consequently, these animals can be regarded as key indicators of Toxoplasma presence in the wild or peri-urban environment. In our study, we examined a total of 174 wild boar meat juice and 128 wolf sera from Italy for the detection of T. gondii IgG using the indirect fluorescent antibody test (IFAT)., Results: The results showed that 40 (22.6%) of the wild boar meat juice and 34 (26.6%) of the wolf serum samples tested positive. Interestingly, there were no significant differences in seropositivity with respect to gender, age group, or the region of origin in both species., Conclusions: Overall the results indicate a moderate exposure in both the species under investigation, highlighting the spread of T. gondii in sylvatic and periurban environments. The prevalence of T. gondii in wild boar is consistent with findings from other studies conducted in Europe. Our study, with a considerably larger sample size compared to the available research in European context, provides valuable data on the seroprevalence of T. gondii in wolves., (© 2024. The Author(s).)

Background: The only natural hosts of Pseudorabies virus (PRV) are members of the family Suidae (Sus scrofa scrofa). In mammals, the infection is usually fatal and typically causes serious neurologic disease. This study describes four Aujeszky's disease cases in free-ranging Italian wolves (Canis lupus italicus). In Italy, the wolf is a strictly protected species and is in demographic expansion., Case Presentation: Three wolves (Wolf A, B, and C) were found in a regional park in Northern Italy, and one (Wolf D) was found in Central Italy. Wolf A and D were alive at the time of the finding and exhibited a fatal infection with epileptic seizures and dyspnoea, dying after a few hours. Wolf B presented scratching lesions under the chin and a detachment of the right earlobe, whilst Wolf C was partially eaten. The wolves showed hepatic congestion, diffuse enteritis, moderate pericardial effusion, severe bilateral pneumonia, and diffuse hyperaemia in the brain. The diagnostic examinations included virological analyses and detection of toxic molecules able to cause serious neurological signs. All four wolves tested positive for pseudorabies virus (PrV). The analysed sequences were placed in Italian clade 1, which is divided into two subclades, "a" and "b". The sequences of Wolf A, B, and C were closely related to other Italian sequences in the subclade b, originally obtained from wild boars and hunting dogs. The sequence from Wolf D was located within the same clade and was closely related to the French hunting dog sequences belonging to group 4., Conclusion: Results showed the presence of PrV strains currently circulating in wild boars and free-ranging Italian wolves. The genetic characterisation of the PrV UL44 sequences from the four wolves confirmed the close relationship with the sequences from wild boars and hunting dogs. This fact supports a possible epidemiological link with the high PrV presence in wild boars and the possibility of infection in wolves through consumption of infected wild boar carcasses or indirect transmission. To the best of our knowledge, this study is the first detection of Pseudorabies virus in free-ranging Italian wolves in northern and central Italy., (© 2023. The Author(s).)

Although it has been established that population density can contribute to the outbreak of the COVID-19 virus, there is no evidence to suggest that economic activities, which imply a significant change in mobility, played a causal role in the unfolding of the pandemic. In this paper, we exploit the particular situation of Sardinia (Italy) in 2020 to examine how changes in mobility due to tourism inflows (a proxy of economic activities) influenced the development of the COVID-19 pandemic. Using a difference-in-differences approach, we identify a strong causal relationship between tourism flows and the emergence of COVID-19 cases in Sardinia. We estimate the elasticity of COVID-19 cases in relation to the share of tourists to be 4.1%, which increases to 5.1% when excluding local residents. Our analysis suggests that, in the absence of tools preventing the spread of infection, changes in population density due to economic activities trigger the pandemic spreading in previously unaffected locations. This work contributes to the debate on the complex relationship between COVID-19 and the characteristics of locations by providing helpful evidence for risk-prevention policies., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Author(s). Published by Elsevier B.V. All rights reserved.)

West Nile (WNV) and Usutu (USUV) viruses are two mosquito-borne viruses belonging to the family Flaviviridae and genus Flavivirus. The natural transmission cycle of WNV and USUV involves mosquitoes and birds, while mammals are thought to be accidental hosts. The goal of this study was to report-in the context of "off-season monitoring" and passive surveillance-the detection of WNV and USUV RNA in wild birds. To this end, we analyzed biological samples of wild birds in Northern Italy, from October to May, hence outside of the regional monitoring period (June-September). The virological investigations for the detection of USUV and WNV RNA were performed using real-time PCR on frozen samples of the brain, myocardium, kidney, and spleen. In a total sample of 164 wild birds belonging to 27 different species, sequences of both viruses were detected: four birds (2.44%) were positive for WNV and five (3.05%) for USUV. Off-season infections of WNV and especially USUV are still widely discussed and only a few studies have been published to date. To the best of our knowledge, this study is the first report on the detection of USUV RNA until December 22nd. Although further studies are required, our results confirm the viral circulation out-of-season of Flavivirus in wild birds, suggesting reconsidering the epidemiological monitoring period based on each individual climate zone and taking into consideration global warming which will play an important role in the epidemiology of vector-borne diseases.

The gray wolf (Canis lupus) expanded its distribution in Europe over the last few decades. To better understand the extent to which wolves could re-occupy their historical range, it is important to test if anthropization can affect their fitness-related traits. After having accounted for ecologically relevant confounders, we assessed how anthropization influenced i) the growth of wolves during their first year of age (n = 53), ii) sexual dimorphism between male and female adult wolves (n = 121), in a sample of individuals that had been found dead in Italy between 1999 and 2021. Wolves in anthropized areas have a smaller overall variation in their body mass, during their first year of age. Because they already have slightly higher body weight at 3-5 months, possibly due to the availability of human-derived food sources. The difference in the body weight of adult females and males slightly increases with anthropization. However, this happens because of an increase in the body mass of males only, possibly due to sex-specific differences in dispersal and/or to "dispersal phenotypes". Anthropization in Italy does not seem to have any clear, nor large, effect on the body mass of wolves. As body mass is in turn linked to important processes, like survival and reproduction, our findings indicates that wolves could potentially re-occupy most of their historical range in Europe, as anthropized landscapes do not seem to constrain such of an important life-history trait. Wolf management could therefore be needed across vast spatial scales and in anthropized areas prone to social conflicts., Competing Interests: The authors declare no competing interest, regarding the study., (Copyright: © 2023 Cerri et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

We report the detection of canine adenovirus type 1 DNA by real-time PCR technique in an oral sample of an Italian wolf ( Canis lupus italicus ). Genetic characterization of the virus revealed a strict relationship with viruses detected in dogs ( Canis lupus familiaris ), wolves, and red foxes ( Vulpes vulpes ), suggesting that transmission between wild animals and dogs had occurred.

Swine influenza viruses (SIVs) have been repeatedly demonstrated to circulate in wild boar ( Sus scrofa) populations, whereas no evidence of exposure to avian influenza viruses (AIVs) has been described in wild boar. To better understand how different environments may influence the ecology of influenza A viruses (IAVs) in wild suid populations, we examined biologic samples of wild boars from two study areas represented by an upland (UL) and a wetland (WL) in northern and central Italy, respectively. Serum samples were collected from 388 wild boars sampled in the UL, whereas both a serum sample and a nasal swab were obtained from each of 35 wild boars sampled in the WL. Twenty of 388 (5.2%) sera from the UL were positive by enzyme-linked immunosorbent assay for the presence of antibodies against influenza A nucleoprotein and some of these samples showed antibodies by hemagglutination inhibition to SIVs of H1N1 (1/20), H1N2 (10/20), and H3N2 (1/20) antigenic subtypes. No IAV-seropositive wild boar was detected in the WL, although one of 35 animals was found to be IAV-positive by both a reverse transcriptase PCR and a real-time reverse transcriptase PCR. We hypothesize an SIV exposure for IAV-seropositive wild boars occupying the UL, whereas a possible AIV spillover from aquatic bird species-natural reservoirs of IAVs-to wild boars in the WL cannot be ruled out. Further research is needed to better understand the role played by wild boars in IAV ecology in Mediterranean habitats.

We investigated the Chlamydia spp. occurrence in Eurasian Collared Doves (Streptopelia decaocto) from urban and suburban areas in northern Italy. Among 76 doves screened, prevalence of Chlamydia spp. was 61%. Chlamydia psittaci genotype E was identified in 33 of the 46 positive samples. The multilocus sequence typing pattern of one highly positive sample showed a new allelic combination. The same molecular features were observed in a C. psittaci strain subsequently isolated from a live dove. Our results reveal a high C. psittaci prevalence in S. decaocto. The spread of this zoonotic pathogen from collared doves to other birds or humans seems to be a potential risk.

Data about the presence of Arcobacter in wild birds are currently lacking. In this study cloacal swabs from 95 collared doves (Streptopelia decaocto), submitted to the Department of Veterinary Medical Sciences (Bologna, Italy) between 2011 and 2013 from various urban and suburban areas of the Emilia-Romagna region (Northern Italy), were tested for the presence of Arcobacter sp. by a rRNA 23S nested Polymerase Chain Reaction (PCR). Eighteen out of 95 (19%) samples showed the expected PCR product. Further cultural and molecular studies are needed to assess the Arcobacter prevalence in wild birds and elucidate their potential epidemiological role as source of animal and human infections.

The present article analyses the current situation of End-of-Life-of-Vehicles (ELVs) management in Europe, with particular attention on Italian condition. Similarly to other European countries, metal recycling is the main activity of the whole system, but such situation is evolving due to the 2000/53/EC Directive, which sets out targets for Reuse, Recycling and Recovery of ELVs. Due to the relevance of the ELVs problem, in 2008 Italian Ministry of Environment subscribed a framework agreement with competent stakeholders as carmakers, dismantlers, shredders. The main result is an industrial plan to promote (amongst other objectives) technological progress for residual waste (Automotive Shredder Residue-ASR) treatment. According with Italian Trial 2006 analysis about ELVs, Reuse and Recycling rate is currently estimated to be about 81%. At the present time, dismantling plants constitute the first collection points for ELVs; for this reason, during 2009 an investigation has been done over a number of ten Authorized Treatment Facilities (ATFs) operating in Italy. The first step of the analysis was aimed to find out major practices and methods through observations of ATFs activities and interviews to operators. Furthermore, the depollution and dismantling treatments of about 70 different ELVs have been observed and timed in detail over a period of three months. The results included the identification of most relevant critical issues in ELVs treatment, such as distortions between scrapping activities and Directive's regulation, and the assessment of the time and of the resources needed to perform each operation. In the second step of the survey, a process simulation model has been built on the basis of such data. The model was aimed to include the real variability and all the uncertainties that are typical of dismantling activities; it is intended as a tool for process layout planning and for its management. Some control parameters have been introduced; these are able to dynamically modify process path depending on ELVs queues and priorities. The model can also be used for the economic assessments of single operations or of the whole treatment activity., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Conjunctival swabs from 44 free-living wild boars culled during a demographic control programme applied in a Regional Park located in the Northern Italy were examined by 16S rRNA encoding gene nested PCR. In total, 22 (50%) wild boars were PCR positive. Sequencing of the amplicons identified Chlamydia suis and Chlamydia pecorum in 12 and 5 samples, respectively. For one sample found PCR positive, the nucleotide sequence could not be determined. Four conjunctival samples showed ≥ 92% sequence similarities to 16S rRNA sequences from Chlamydia-like organisms, as did large intestine, uterus, and vaginal swabs from the same four animals. Amoeba DNA was found in one Chlamydia-like organism positive conjunctival swab. To our knowledge, this is the first detection of members of the Parachlamydiaceae family in wild boars, confirming a large animal host range for Chlamydia-like organisms.

Sardinia is a Mediterranean island with a long geological history, leading to a separation process from continental Europe during the Miocene. As a consequence, in this insular habitat some wild bird species developed endemic forms, some of which are currently threatened. The aim of this study is to evaluate the possible animal health risk associated with a potential avian influenza virus (AIV) circulation in Sardinian wild bird populations. Overall, 147 cloacal swabs were sampled in the Sardinia region from June 2009 to September 2011. Samples were obtained from 12 taxonomic orders, including 16 families and 40 species of birds. Based on the endangered host status or on the ecology of the host-virus interaction, samples were categorized into three groups of species: 1) endemic, endangered, or both (17 species); 2) potential reservoir (21 species); and 3) potential spillover (two species). Cloacal swabs were tested by reverse transcription (RT)-PCR for influenza A virus matrix gene amplification. Forty-one serum samples were tested by nucleoprotein-enzyme-linked immunosorbent assay (NP-ELISA) for antibodies against influenza A virus nucleoprotein and by hemagglutination inhibition assay for detection of seropositivity against H5 and H7 AIV subtypes. No cloacal swabs tested RT-PCR positive for AIV, whereas two weak seropositive results were detected by NP-ELISA in a mallard (Anas platyrhynchos) and in a yellow-legged gull (Larus michahellis). The low or absent AIV circulation detected in Sardinia's wild birds during the study suggests a naïve status in these avian populations. These data provide new information on AIV circulation in Sardinia's wild birds that could be applied to implement conservation strategies for threatened species.

Hepatitis E virus (HEV) is the causative agent of Hepatitis E. Swine and human HEV strains are genetically related, suggesting the occurrence of zoonotic transmission. Recently, in Japan, cases of food-borne HEV transmission have been described in people after consuming raw or undercooked meat from wild boars or pigs. Although, swine HEV strains have been detected in pig herds in many European countries, only minimal information is presently available about the circulation and the prevalence of HEV in wild boars in Europe. In this study, we investigated the presence of HEV in a demographic managed wild boar population in Italy. Detection of HEV RNA was accomplished using a nested reverse-transcription polymerase chain reaction on bile samples from 88 shot animals. HEV RNA was detected in 22 out of 88 animals tested (25%). Phylogenetic analysis on the nucleotide sequences obtained from 10 positive PCR products indicated that only one HEV strain was circulating in the wild boar population considered, and that this strain was closer to human and swine HEV strains circulating in Europe than to wild boar Japanese strains.

Two epidemics of avian influenza due to H5 and H7 highly pathogenic viruses occurred in poultry in Italy in 1997/98 and 1999/2000, respectively. The circulation of these serotypes in wild aquatic birds was investigated examining 638 cloacal swabs and 621 sera collected from 150 gulls, 162 coots, and 326 ducks trapped in Italian wetlands from 1998 to 2000. Seroprevalences against influenza A viruses, detected by a double-antibody sandwich-blocking enzyme-linked immunosorbent assay (ELISA), were 11% in gulls, 16% in coots, and 45% in ducks. Among the Anatidae group, duck species wintering in Mediterranean areas showed significantly higher values than ducks wintering in South-Saharan areas of Africa. In order to detect H5 and H7 antibodies, the haemagglutination-inhibition assay and two competitive ELISA tests (H5-ELISA and H7-ELISA) using monoclonal antibodies specific for H5 and H7 subtypes were performed. None of the aquatic bird species were found seropositive for H7 subtype, whereas H5-positive sera were found by both the haemagglutination-inhibition and ELISA assays in ducks only. The highest H5 seroprevalences were detected by H5-ELISA; overall, 5% (10/201) of duck species wintering in Mediterranean areas tested positive by this assay, with annual seroprevalences ranging from 2% (2/123) to 12% (6/51). In the present study, only five viruses belonging to H1N1, H11N6, and H2N3 subtypes were isolated from ducks. However, the H5 seroconversion observed in one mallard duck at the beginning of 1998 indicates that H5 virus circulation also occurred in the study area.

Since the "bird flu" incident in Hong Kong SAR in 1997, several studies have highlighted the substantial role of domestic birds, such as turkeys and chickens, in the ecology of influenza A viruses. Even if recent evidence suggests that chickens can maintain several influenza serotypes, avian influenza viruses (AIVs) circulating in domestic species are believed to be introduced each time from the wild bird reservoir. However, so far the direct precursor of influenza viruses from domestic birds has never been identified. In this report, we describe the antigenic and genetic characterization of the surface proteins of H7N3 viruses isolated from wild ducks in Italy in 2001 in comparison to H7N3 strains that circulated in Italian turkeys in 2002-2003. The wild and domestic avian strains appeared strictly related at both phenotypic and genetic level: homology percentages in seven of their genes were comprised between 99.8% (for PB2) and 99.1% (for M), and their NA genes differed mainly because of a 23-aminoacid deletion in the NA stalk. Outside this region of the molecule, the NAs of the two virus groups showed 99% similarity. These findings indicate that turkey H7N3 viruses were derived "in toto" from avian influenza strains circulating in wild waterfowl 1 year earlier, and represent an important step towards the comprehension of the mechanisms leading to interspecies transmission and emergence of potentially pandemic influenza viruses.

The mechanisms of perpetuation of influenza A viruses in aquatic birds, their main reservoir in nature, have not yet been completely clarified. One hypothesis is that they continue to circulate in waterfowl throughout the year, even though virus isolations during the winter months are rare. We analyzed influenza virus circulation in wild ducks in Italy during six winter seasons (1993-99), using virus isolations and serological analyses. It was apparent that influenza A viruses were constantly circulating in wild birds during all the seasons considered. Moreover, seroconversion rates (obtained from ducks recaptured during the same season) suggest a frequency of influenza infections higher than expected on the basis of the virus isolation rates.