Development of a qPCR Duplex Assay for simultaneous detection of Fascioloides magna and Galba truncatula in eDNA samples: Monitoring beyond boundaries.

Parasites constitute a significant economic burden and highly impact environmental, public, and animal health. The emergence of many parasitic diseases is environmentally mediated and they share the same biogeography with humans and both domestic and wild animals. American liver fluke, Fascioloides magna - a trematode parasite of domestic and wild ungulates - is an example of the anthropogenic introduction of an "invasive alien species" in Italy and Europe. Multiple introductions to Europe have led to the biogeographical expansion of the parasite across the Danube region mainly provided by the presence of suitable habitats for all hosts involved in the parasite's life cycle, human-assisted transport, and drastic environmental events such as flooding. In Italy, it was introduced and established in La Mandria Regional Park (LMRP) near Turin in 1865 along with imported wapitis (Cervus elaphus canadensis) from North America (Bassi, 1875), but with no reported expansion to the surrounding areas. LMRP isolated F. magna focus, poses an important threat of possible expansion since the enclosed area is vulnerable to occasional bidirectional passage of roe deer. Additionally, tributary rivers to the Po river system, traversing the enclosed area, could further bolster the possibility of such spread. In this study, we developed a duplex qPCR assay for F. magna and its principal intermediate host Galba truncatula optimized for testing eDNA samples to meet the needs for surveillance of the parasite. Moreover, we validated the developed assay in natura by testing samples derived from filtered water and sediments collected inside and outside LMRP's fenced-off area. Our findings for the first time demonstrate the presence of F. magna's eDNA outside the park's internal fenced-off area.

How abundant is a species at the limit of its distribution range? Crested porcupine Hystrix cristata and its northern population.

The crested porcupine (Hystrix cristata) is a rodent present in Africa and southern Europe (Italy exclusively). The Italian population is expanding from the centre to the north and south, but little is known about the species' abundance. Reliable population density estimates are important for monitoring trends in wildlife populations and for developing effective conservation and management strategies. In this context, we aimed to first report crested porcupine population density on the northern limit of its current distribution range using a non-invasive approach. Specifically, we randomly placed 38 camera traps in an area of 242 km2 in north Italy (Lombardy region), and we applied camera trap distance sampling. We estimated a porcupine density of 0.49 ind·km-2 (±0.33, standard error). The results presented here are the first crested porcupine density estimate accounting for imperfect detection (i.e. species present but not detected). The abundance estimate reported here is fundamental for a better understanding of the species status in Europe and for implementing conservation and management plans.

The contribution of citizen science in the surveillance of wildlife and related arthropods.

Environmental and anthropogenic factors may significantly affect the diffusion of wild animals, enhancing the interface of human­wildlife interactions and driving the spread of pathogens and vector-borne diseases between animals and humans. However, in the last decade, the involvement of citizens in scientific research (the so-called citizen science approach, henceforth abbreviated as CS) provided a network of large-scale and cost-effective surveillance programmes of wildlife populations and their related arthropod species. Therefore, this review aims to illustrate different methods and tools used in CS studies, by arguing the main advantages and considering the limitations of this approach. The CS approach has proven to be an effective method for establishing density and distribution of several wild animal species, in urban, peri-urban and rural environments, as well a source of information regarding vector­host associations between arthropods and wildlife. Extensive efforts are recommended to motivate citizens to be involved in scientific projects to improve both their and our knowledge of the ecology and diseases of wildlife. Following the One Health paradigm, collaborative and multidisciplinary models for the surveillance of wildlife and related arthropod species should be further developed by harnessing the potentiality of the CS approach.

The Dangerous Side of Being a Predator: Toxoplasma gondii and Neospora caninum in Birds of Prey.

Toxoplasma gondii and Neospora caninum are apicomplexan protozoa of major concern in livestock and T. gondii is also considered one of the major threats and a public health concern. These protozoa have a wide range of intermediate hosts, including birds. The present work aimed to assess the prevalence of these cyst-forming parasites in migratory and sedentary birds of prey. The skeletal muscle and myocardium of 159 birds of prey from Central Italy, belonging to 19 species and recovered across 6 Wildlife Recovery Centers/Care structures along the Italian migratory route, were collected specifically for molecular (PCR) and for histopathological analysis to detect T. gondii and N. caninum. For the molecular analysis, genomic DNA was extracted. The DNA was tested by sequence typing, targeting GRA6, 529 bp repeated element, B1, PK1, BTUB, SAG2, alt.SAG2, and APICO genes for T. gondii and to end-point PCR targeting NC5 gene for N. caninum. Thirty-seven out of the one hundred and fifty-nine analyzed samples tested positive for T. gondii with a prevalence of 23.27% and nine for N. caninum, with a prevalence of 5.66%. Thirty-two sequences were obtained from the thirty-seven isolates of T. gondii. Among these, 26 presented alleles compatible with type I strain in 1 or more loci, 4 with type II strain and 2 consisted of atypical strains. Toxoplasma gondii genetic variability in birds of prey confirms previous findings of wildlife as reservoirs of atypical strains. Results from the histology showed few protozoal tissue cysts in skeletal muscle (n. 4) and hearts (n. 2).

Fifty Years of Wildlife Diseases in Europe: A Citation Database Meta-Analysis.

Although wildlife has progressively been recognized as a booster for the spillover of pathogens to humans and other wild and domestic animals, the interest of scientists in this topic has not been constant over time and uniform in its targets. Epidemiological outbreaks and technological development have contributed to this. Through the analysis of the number of publications from a citation database, we aimed to obtain an indicator of the scientific community's interest towards wildlife diseases over the years. Using Web of Science, bibliographic searches were performed by adding to the basic search string "Wildlife AND Disease" terms detailing topics such as aspect investigated, pathogen type, aetiologic group and species group. For each host species group, the 100 most frequent Medical Subject Headings (MeSHs) related to specific diseases in each decade were extracted. The scientific production regarding wildlife diseases has increased 3.7 times the relative proportion of publications on diseases during the last 50 years, focusing mainly on zoonotic or epizootic pathogens, and with a sharp growth in contemporary hot topics. Wildlife disease scenarios are complex and challenging to approach. Knowing the trends in the scientific interest in the past decades may pose a guide to direct future steps and actions in several fields, from public health to ecosystem management.