Search Results (28)

Highly pathogenic avian influenza (HPAI) H5N1 continues to pose a major threat to animal and public health. Since its emergence, clade 2.3.4.4b has become the dominant global lineage, associated with widespread outbreaks in Europe. This study aimed to characterise the H5N1 strains detected in backyard chickens in Serbia and investigate their phylogenetic and phylogeographic relationships with historical and currently circulating strains in Serbia and the broader region. Samples collected in 2024 were tested by qRT-PCR, and positive samples were propagated in MDCK cells. Whole genome sequencing of isolated virus was performed using the MinION Mk1C platform (ONT, Oxford, UK). Bayesian phylogenetic and phylogeographic analyses were conducted using BEAST X and SPREAD3 v.9.6. The results revealed three independent introductions of H5N1 into Serbia between 2020 and 2024. The spatiotemporal diffusion patterns confirmed both north–south and west–east viral spread from Northern and Central Europe into the Balkans. Additionally, Serbia was identified as a critical transit and divergence point in the regional transmission network, highlighting its role in the spread of H5N1 between Western and Eastern Europe. These findings underscore the importance of continued genomic surveillance in both domestic and wild bird populations to better understand and reduce HPAI spread. Full article

Birds are hosts to a diverse assemblage of vector-transmitted haemosporidian parasites. However, the true genetic diversity and many host–parasite interactions are still unknown, particularly in under-represented species groups such as wild doves and pigeons (Columbiformes). In this study, we examined the prevalence and lineage diversity of haemosporidian genera Plasmodium, Leucocytozoon, and Haemoproteus in three species of wild columbids, sampled in Germany. Examinations were performed by applying molecular methods (nested PCR and one-step multiplex PCR) and blood smear examination, and their respective advantages and disadvantages are discussed. In the case of the European Turtle Dove Streptopelia turtur, samples were collected along a west–east gradient throughout Germany, covering migratory birds from the Western and Central-Eastern flyway of this species. Although no infection was detected in the Stock Dove Columba oenas samples, 53% of Turtle Dove and 86% of Common Woodpigeon Columba palumbus harbored a parasite of at least one haemosporidian genus, revealing previously unknown lineage–host interactions. We were not able to demonstrate a correlation between infection status (presence or absence of infection based on PCR results) and parasitemia with condition based on the heterophil to lymphocyte ratio (H/L ratio). Neither lineage occurrence nor prevalence of infection followed any geographically specific patterns. Thus, haemosporidian lineages found in Turtle Doves could not be used as a marker of geographic origin that would allow the tracking of their nonbreeding distribution. Full article

Avian influenza (AI), caused by Alphainfluenzavirus (family Orthomyxoviridae), poses significant threats to poultry, biodiversity, and public health. AI outbreaks in poultry lead to severe economic losses, while highly pathogenic strains (HPAIVs) severely impact wild bird populations, with implications for biodiversity and potential zoonotic risks. Similarly, arboviruses such as West Nile virus (WNV) and Usutu virus (USUV) are emerging zoonoses. WNV can cause severe neurological diseases in birds, humans, and other animals, while USUV significantly affects blackbird populations and has zoonotic potential, though human cases remain rare. This study investigated avian viruses in 1654 wild birds from 75 species that died at the Wildlife Rescue Center in Vanzago, Lombardy, during 2023–2024. Necropsies were conducted, and virological analyses were performed to detect avian influenza viruses, WNV, and USUV. Among the tested birds, 15 were positive for H5N1 HPAIV clade 2.3.4.4b, all in 2023, including 13 Chroicocephalus ridibundus, one Coturnix coturnix, and one Columba palumbus. Additionally, 16 tested positive for WNV (15 for lineage 2 and one for lineage 1), one for USUV, and 11 co-infections WNV/USUV were recorded in 2023–2024. These findings underscore the importance of avian viral passive surveillance in identifying epidemiological trends and preventing transmission to other species, including mammals and humans. Full article

African swine fever (ASF), characterized by high mortality rates in infected animals, remains a significant global veterinary and economic concern, due to the widespread distribution of ASF virus (ASFV) genotype II across five continents. In this study, ASFV strains collected in Italy during 2022–2023 from two geographical clusters, North-West (Alessandria) and Calabria, were fully sequenced. In addition, an in vivo experiment in pigs was performed. Complete genomic sequencing of 30 strains revealed large-fragment deletions and translocations. In Alessandria, five samples showed two different deletions in the 5′ genomic region: a ~4340 bp deletion (positions ~9020–13,356 in Georgia 2007/1) in four samples and a 2162 bp deletion (positions 17,837–19,998) in one sample. Another strain showed a truncation of 1950 bp at the 3′ end. In Calabria, strains showed a 5137 bp deletion (positions 10,755–15,891) and ~2 kb truncations in the 3′ region. Two strains showed a translocation from the region 1–2244 to positions 188,631–190,584. In vivo characterization of the deleted strain 22489.4_2312/RC/2023 revealed identical disease progression to the wild-type strain, with severe ASF symptoms in inoculated pigs. This study is the first to report natural deleted strains of ASFV in Italy, revealing unique genomic deletions distinct from those in previously known strains. Full article

Grasspea (Lathyrus sativus L.) is a climate-smart legume crop with adaptation to fragile agroecosystems. The genus Lathyrus is recognized for its vast genetic diversity, encompassing over 160 species, many of which are cultivated for various purposes across different regions of the world. Among these, Lathyrus sativus is widely cultivated as food, feed, and fodder in South Asia, Sub-Saharan Africa, and the Central and West Asia and North Africa (CWANA) regions. Its global cultivation has declined substantially due to the stigma posed by the presence of neurotoxin β-N-oxalyl-L-α, β-diaminopropionic acid (β-ODAP) in its seeds and green foliage. Overconsumption for a longer period of grasspea seeds harvested from landraces may lead to a neurological disorder called neurolathyrism in humans. ODAP is an obstacle for grasspea expansion, but crop wild relatives (CWRs) have been found to offer a solution. The incorporation of CWRs, particularly Lathyrus cicera, and landraces into breeding programs may reduce the ODAP content in grasspea varieties to a safer level. Recent advances in genomics-assisted breeding have expanded the potential for utilizing challenging CWRs to develop grasspea varieties that combine ultra-low ODAP levels with improved yield, stability, and adaptability. Further progress in omics technologies—such as transcriptomics, proteomics, and metabolomics—along with genome sequencing and editing, has greatly accelerated the development of grasspea varieties with reduced or zero ODAP content, while also enhancing the plant’s agronomic value. This review highlights the significance of utilizing CWRs in pre-breeding programs, and harnessing advanced tools and technologies to enhance the performance, adaptability, and resilience of grasspea in response to changing environmental conditions. Full article

In Italy, there are hundreds of research, monitoring, and surveillance activities targeting emerging and re-emerging pathogens. These activities heavily rely on hunters for sample collection and early identification of morbidity/mortality events. The objective of this review is to describe and quantify the contribution of hunters in the context of disease research, monitoring, and surveillance in wild populations. A literature review and descriptive summary statistics were performed following PRISMA-2020 guidelines; articles were obtained from major scientific databases, abstracts from national and international conferences, proceedings, graduate-level theses from online library repositories, and direct contact with academic experts. The contribution of hunters in terms of sample collection for health-related activities on wildlife amounts to 400,000 sampled animals. Wild boars were involved in 158 surveillance systems/research studies, followed by red deer (71), foxes (63), and roe deer (59). The pathogens under surveillance were mainly zoonotic (Salmonella spp.), emerging (Hepatitis E virus), and/or vector-borne (West Nile virus). The temporal distribution of scientific papers followed a positive trend that reflects the growing interest in wildlife from different sectors. These results highlight how wildlife health-related efforts are a paradigm of the concept of One Health, in which the role of hunters is crucial to ensure sample availability, and it constitutes the base of much current wildlife health research, monitoring, and surveillance. Full article

(1) Background: Bacillus cereus biovar anthracis (Bcbva) was the causative agent of an anthrax-like fatal disease among wild chimpanzees in 2001 in Côte d’Ivoire. Before this, there had not been any description of an anthrax-like disease caused by typically avirulent Bacillus cereus. Genetic analysis found that B. cereus had acquired two anthrax-like plasmids, one a pXO1-like toxin producing plasmid and the other a pXO2-like plasmid encoding capsule. Bcbva caused animal fatalities in Cameroon, Democratic Republic of Congo, and the Central African Republic between 2004 and 2012. (2) Methods: The pathogen had acquired plasmids in the wild and that was discovered as the cause of widespread animal fatalities in the early 2000s. Primate bones had been shipped out of the endemic zone for anthropological studies prior to the realized danger of contamination with Bcbva. Spores were isolated from the bone fragments and positively identified as Bcbva. Strains were characterized by classical microbiological methods and qPCR. Four new Bcbva isolates were whole-genome sequenced. Chromosomal and plasmid phylogenomic analysis was performed to provide temporal and spatial context to these new strains and previously sequenced Bcbva. Tau and principal component analyses were utilized to identify genetic and spatial case patterns in the Taï National Park anthrax zone. (3) Results: Preliminary studies positively identified Bcbva presence in several archival bone fragments. The animals in question died between 1994 and 2010. Previously, the earliest archival strains of Bcbva were identified in 1996. Though the pathogen has a homogeneous genome, spatial analyses of a subset of mappable isolates from Taï National Park revealed strains found closer together were generally more similar, with strains from chimpanzees and duikers having the widest distribution. Ancestral strains were located mostly in the west of the park and had lower spatial clustering compared to more recent isolates, indicating a local increase in genetic diversity of Bcbva in the park over space and time. Global clustering analysis indicates patterns of genetic diversity and distance are shared between the ancestral and more recently isolated type strains. (4) Conclusions: Our strains have the potential to unveil historical genomic information not available elsewhere. This information sheds light on the evolution and emergence of a dangerous anthrax-causing pathogen. Full article

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. Full article

Dioscorea praehensilis Benth. is a semi-wild yam species and a valuable source of resistance trait genes. To access the agronomic and tuber quality performance, eleven quantitative phenotypic traits were used to discriminate and identify promising accessions among 162 accessions of D. praehensilis collected in Ghana. Significant and high genetic variability (p < 0.001) for all eleven quantitative traits was found among the evaluated accessions. Moderate broad-sense heritability (H²) (30–60%) was observed for all the evaluated quantitative traits except the response to YMV and tuber hardness. The accessions were clustered into three groups; each cluster displayed genotypes with good potentiality for the different traits evaluated. Path coefficient analysis revealed positive contributions (p < 0.01) of the number of tubers per plant, tuber length, tuber width, stem internode length, number of internodes, and tuber flesh hardness to the total tuber weight per plant. Through the multi-trait genotype–ideotype distance index (MGIDI), 24 accessions were identified from the 162 evaluated accessions as top-ranking and could be used as progenitors for trait introgression. The results of this study provide insight for future yam breeding and improvement programs in West Africa. Full article

Tick-borne encephalitis virus (TBEV) is a flavivirus transmitted by ticks. Serological screenings in animals are performed to estimate the prevalence and distribution of TBEV. Most screenings consist of a primary screening by ELISA, followed by confirmation of positive samples by plaque reduction neutralization tests (PRNTs). In this study, 406 wild boar sera were tested with 2 regularly used commercial ELISAs for flavivirus screening in animals (Immunozym FSME (TBEV) IgG All Species (Progen) and ID Screen West Nile Competition (Innovative Diagnostics)) and PRNTs for TBEV and USUTU virus. The results showed that the Immunozym and IDScreen ELISAs had low relative sensitivities of 23% and 20%, respectively, compared to the PRNT results. The relative specificities were 88% and 84% due to cross reactions with USUTU virus-specific antibodies. The minimal TBEV prevalence in our sample set was 8.6% when determined by PRNT. When the screening approach of ELISA testing followed by PRNT confirmation was applied, a TBEV seroprevalence of only 2.0% and 1.7% was found. The suboptimal performance of the ELISAs was confirmed by testing sera collected from experimentally TBEV-infected sheep. While the PRNT detected TBEV specific antibodies in 94% of samples collected between 7 and 18 days post-infection, the ELISAs classified only 50% and 31% of the samples as positive. Both routinely used ELISAs for TBEV antibody screening in animal sera were shown to have a low sensitivity, potentially leading to an underestimation of the true prevalence, and furthermore cross-react with other flavivirus antibodies. Full article

Abies nebrodensis (Lojac.) Mattei is an endemic species of the north-west of Sicily located in an 84 ha area in the Madonie Regional park. The current population is limited to 30 relic adult trees and a fluctuating number of juveniles of natural regeneration. The species is defined as “Critically Endangered” in the Italian list of threatened plants and is classified as CR-D in the 2000 IUCN Red List of Threatened Species. This article reports the key action undertaken by the LIFE4FIR project aimed at preserving A. nebrodensis, and the results obtained so far in three years of activity. OpenArrays SNPs genotyping revealed a high rate of inbreeding in the natural population and that the adult trees are genetically related. Controlled cross-pollination was consequently performed to increase the genetic variability of the progeny. Outbred offspring are currently being grown in the nursery. Reforestation has been planned by using 4000 selected outbred seedlings in 10 areas within Madonie Park to create re-diffusion cores. Support and protection of the relic population have been implemented through regular phytosanitary surveys, as well as new fencing and video surveillance systems against grazing and wild herbivores. A seedbank and cryobank for the long-term germplasm conservation have been established. Full article

Flavivirus outbreaks require fast and reliable diagnostics that can be easily adapted to newly emerging and re-emerging flaviviruses. Due to the serological cross-reactivity among flavivirus antibodies, neutralization tests (NT) are considered the gold standard for sero-diagnostics. Here, we first established wild-type single-round infectious virus replicon particles (VRPs) by packaging a yellow fever virus (YFV) replicon expressing Gaussia luciferase (Gluc) with YFV structural proteins in trans using a double subgenomic Sindbis virus (SINV) replicon. The latter expressed the YFV envelope proteins prME via the first SINV subgenomic promoter and the capsid protein via a second subgenomic SINV promoter. VRPs were produced upon co-electroporation of replicon and packaging RNA. Introduction of single restriction enzyme sites in the packaging construct flanking the prME sequence easily allowed to exchange the prME moiety resulting in chimeric VRPs that have the surface proteins of other flaviviruses including dengue virus 1–-4, Zika virus, West Nile virus, and tick-borne encephalitis virus. Besides comparing the YF-VRP based NT assay to a YF reporter virus NT assay, we analyzed the neutralization efficiencies of different human anti-flavivirus sera or a monoclonal antibody against all established VRPs. The assays were performed in a 96-well high-throughput format setting with Gluc as readout in comparison to classical plaque reduction NTs indicating that the VRP-based NT assays are suitable for high-throughput analyses of neutralizing flavivirus antibodies. Full article

Wild birds play an important role in the circulation and spread of pathogens that are potentially zoonotic or of high economic impact on zootechnical production. They include, for example, West Nile virus (WNV), Usutu virus (USUV), avian influenza virus (AIV), and Newcastle disease virus (NDV), which, despite having mostly an asymptomatic course in wild birds, have a strong impact on public health and zootechnical production. This study investigated the presence of these viruses in several wild bird species from North Italy during the biennium 2019–2020. Wild birds derived from 76 different species belonging to 20 orders. Out of 679 birds, 27 were positive for WNV (lineage 2) with a prevalence of 4%; all birds were negative for USUV; one gull was positive for H13N6 influenza virus, and 12 samples were positive for NDV with a prevalence of 2%. Despite the low prevalence observed, the analyses performed on these species provide further data, allowing a better understanding of the diffusion and evolution of diseases of both economic and zoonotic importance. Full article

The mission of botanical gardens is to work towards the conservation and sustainable use of plant diversity. In the case of gardens established on the premises of former manor estates, actions are also being taken to properly manage existing natural resources, e.g., historic tree stands. This article is based on the case study of the Botanical Garden in Lublin. To assess the health condition of the trees growing in the oldest part of the park—Kościuszko Redoubt—sonic tomography examinations were performed. The article presents the practical application of a newer form of digital imaging, put to the service of assessing the health of old trees in an important public space. The Redoubt is the only work of defense architecture in Poland related to the activity of General Tadeusz Kościuszko, the hero of Poland and the United States of America. It has survived in an excellent condition, preserved without significant changes, as an earth structure with ramparts; an interior, i.e., a yard; a cannon post located on an inaccessible cliff; and a brick basement serving as a shelter for soldiers and a warehouse. The entire surroundings also survived: ravines masked with a wild green, embankments with a safe hidden access road to the Redoubt, and a shaped defense line. The Botanical Garden area is located in the Sławinek district, in the valley of the Czechówka River, and includes a fragment of it and the slope adjacent to the west, cut by three loess ravines. The Botanical Garden was established in 1965, where there was a manor farm earlier, and then in the 19th century, a spa that was destroyed during World War I and II. Good soils and a varied microclimate provide the right ecological conditions for the high biodiversity of plants. With an area of 21.15 ha, the Garden area has the character of a landscape park with the theme of a manor garden from the turn of the 18th/19th century and a spa garden. The primary purpose of the work was to investigate the effectiveness of the use of computer tools in the sustainable management of historical greenery in the Botanical Garden. Research has shown that diagnostics with the use of sound waves allow for an accurate diagnosis and quick protective measures against the tree, improving the safety of visitors to the botanical garden. Moreover, the obtained results and a historical query were used to prepare an application to recognize Redoubt as a monument. Full article

West Nile virus (WNV) and Usutu virus (USUV) are zoonotic arboviruses. These flaviviruses are mainly maintained in the environment through an enzootic cycle involving mosquitoes and birds. Horses and humans are incidental, dead-end hosts, but can develop severe neurological disorders. Nevertheless, there is little data regarding the involvement of other mammals in the epidemiology of these arboviruses. In this study, we performed a serosurvey to assess exposure to these viruses in captive birds and mammals in a zoo situated in the south of France, an area described for the circulation of these two viruses. A total of 411 samples comprising of 70 species were collected over 16 years from 2003 to 2019. The samples were first tested by a competitive enzyme-linked immunosorbent assay. The positive sera were then tested using virus-specific microneutralization tests against USUV and WNV. USUV seroprevalence in birds was 10 times higher than that of WNV (14.59% versus 1.46%, respectively). Among birds, greater rhea (Rhea Americana) and common peafowl (Pavo cristatus) exhibited the highest USUV seroprevalence. Infections occurred mainly between 2016–2018 corresponding to a period of high circulation of these viruses in Europe. In mammalian species, antibodies against WNV were detected in one dama gazelle (Nanger dama) whereas serological evidence of USUV infection was observed in several Canidae, especially in African wild dogs (Lycaon pictus). Our study helps to better understand the exposure of captive species to WNV and USUV and to identify potential host species to include in surveillance programs in zoos. Full article