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Advances in the epidemiological tracing of pathogen transmission have been largely driven by the increasing characterisation of whole-genome sequence data obtained at a finer resolution from infectious disease outbreaks. Dynamic models that integrate genomic and epidemiological data further enhance inference on the evolutionary history and transmission dynamics of epidemic outbreaks by reconstructing the network of ‘who-infected-whom’. Swine Vesicular Disease (SVD) was present in Italy from 1966 until 2015, and since the mid-1990s, it has mainly been circulating within Italy’s central-southern regions with sporadic incursions to the north of the country. However, a recrudescence of SVD in northern Italy was recorded between November 2006 and October 2007, leading to a large-scale epidemic that significantly affected the intensive pig industry of the Lombardy region. In this study, by using whole-genome sequence data in combination with epidemiological information on disease occurrences, we report a retrospective epidemiological investigation of the 2006–2007 SVD epidemic, providing new insights into the transmission dynamics and evolutionary mode of the two phases that characterised the epidemic event. Our analyses support evidence of undetected premises likely missed in the chain of observed infections, of which the role as the link between the two phases is reinforced by the tempo of SVD virus evolution. These silent transmissions, likely resulting from the gradual loss of a clear SVD clinical manifestation linked to sub-clinical infections, may pose a risk of failure in the early detection of new cases. This study emphasises the power of joint inference schemes based on genomic and epidemiological data integration to inform the transmission dynamics of disease epidemics, ultimately aimed at better disease control.

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.

Lumpy skin disease virus (LSDV), a member of the genus, poses a significant threat to livestock health and productivity in both endemic and newly affected regions. The disease is primarily transmitted by blood-feeding insects, leading to fever, cutaneous nodules, lymphadenopathy, and substantial economic losses. While vaccination remains the cornerstone of control efforts, effective surveillance-especially in high-risk areas-relies on robust and scalable diagnostic tools. Although the virus neutralization test is considered the reference standard among serological assays for detecting neutralising antibodies, it is labor-intensive and requires high-containment laboratories. In this study, we produced and evaluated two recombinant LSDV antigens: ORF074 (p32), a well-known immunodominant protein, and ORF060 (homologous to the Vaccinia virus L1R), a myristoylated membrane protein identified as a promising immunogenic target. Both proteins were expressed in . Recombinant p32 was purified under native conditions, whereas recombinant L1R required denaturation and refolding. The antigens were used to develop two indirect ELISAs, and were evaluated using sera from experimentally infected cattle, as well as both vaccinated and infected field samples from Albania and Serbia. Both assays demonstrated high immunoreactivity and strong concordance with the VNT. These results support the suitability of both antigens for use in serological assays and suggest that a combined, multi-target ELISA approach could enhance diagnostic sensitivity. Once validated for routine use, these novel tools may significantly improve large-scale, cost-effective serological surveillance of LSDV in endemic and at-risk regions.

During the last 25 years, swine vesicular disease (SVD) has occurred in Italy mostly sub-clinically. Therefore, regular testing of fecal samples from suspected holdings and high turnover premises was fundamental to identifying virus circulation and to achieve SVD eradication. In this study, we evaluated diagnostic performances of six genomic amplification methods, using positive fecal samples from 78 different outbreaks (1997–2014), which included different lineages. Comparison of three RT-PCRs, designed to amplify the same 154 nt portion of the gene 3D, demonstrated that a conventional and a real-time based on SYBR Green detection assay showed the highest diagnostic sensitivity, detecting all samples, while a real-time TaqMan-based test missed three cases, owing to two mismatches in the probe target sequence. Diagnostic and analytical specificities were optimal, as 300 negative field samples and other enteroviruses reacted negative. Three further evaluated tests, previously described, were a 3D-targeted reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) and two real-time RT-PCRs targeted on the 5′UTR region. Here, the presence of multiple mismatches in probe and primers reduced the diagnostic performances, and two of the assays were unable to detect viruses from one sub-lineage. These results highlight that the choice of tests using less nucleotide targets significantly contributed to the success of the SVD eradication plan.

Abstract Swine vesicular disease virus (SVDV) was considered endemic in Italy until 2015, when eradication was achieved. From 1992, when the most recent antigenic-genomic variant of SVDV was identified, to 2015, ~685 SVD outbreaks occurred. In order to understand the evolutionary drivers of SVDV in Italy, 152 viral strains were selected and whole-genome sequencing was performed. Comparison of the polyprotein coding region revealed a minimum pairwise identity of 85% and 98% at the nucleotide and amino acid levels, respectively, consistent with the antigenic homogeneity of the analysed strains. Phylogenetic analysis confirmed the presence of two sublineages: sublineage 1, which evolved and circulated specifically in Italy since 1995, and sublineage 2, which also includes strains circulating in both Spain and Portugal in 1993 and in Portugal during 2003. Recombination analysis revealed a breakpoint site within the 2B coding region, resulting in a recombination strain originating from viruses belonging to the two sublineages, which was dated to the beginning of 2008. This single recombination event gave rise to at least 20 recombinant strains, which circulated together with their parental strains until 2010 and from then on circulated on their own until eradication in 2015. Our data showed that, apart from the recombination event, SVDV, which has been present on Italian territory for >20 years, has not been subject to positive selective pressures that would have conferred a possible evolutionary advantage.

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.