Purification of Epizootic Hemorrhagic Disease Virus (and Other Orbiviruses) Particles from Infected Mammalian or Insect Cells.

Epizootic hemorrhagic disease virus (EHDV), like other orbiviruses, infects and replicates in mammalian and insect vector cells. Within its ruminant hosts EHDV, like bluetongue virus (BTV), it has mainly been associated with infection of endothelial cells of capillaries as well as leukocyte subsets. Furthermore, EHDV infects and replicates within its biological vector, Culicoides biting midges and Culicoides-derived cells. A wide range of common laboratory cell lines such as BHK, BSR, and Vero cells are susceptible to infection with certain EHDV strains. Cell culture supernatants of infected cells are commonly used for both in vivo and in vitro infection studies. For specific virological or immunological studies, using highly purified virus particles, however, might be beneficial or even required. Here we describe a purification method for EHDV particles, which had been originally developed for certain strains of BTV.

Involving citizen scientists in monitoring arthropod vectors of human and zoonotic diseases: The case of Mosquito Alert in Italy.

Citizen science has been particularly effective in gathering reliable, timely, large-scale data on the presence and distributions of animal species, including mosquito vectors of human and zoonotic pathogens. This involves the participation of citizen scientists in research projects, with success strongly dependent on the capacity to disseminate project information and engage citizen scientists to contribute their time. Mosquito Alert is a citizen science that aids in the system surveillances of vector mosquitoes. It involves citizen scientists providing expert-validated photos of targeted mosquitoes, along with records of bites and breeding sites. Since 2020 the system has been disseminated throughout Europe. This article uses models to analyze the effect of promotion activities carried out by the Mosquito Alert ITALIA team from October 2020 to December 2022 on the number of citizen scientists recruited and engaged in the project, and their performance in mosquito identification. Results show a high level of citizen scientist recruitment (N > 18.000; 37 % of overall European participants). This was achieved mostly through articles generated by ad hoc press releases detailing the app's goals and functioning. Press releases were more effective when carried out at the beginning and end of the mosquito season and when mosquito's public health significance was emphasized. Despite the high number of records received (N > 20.000), only 30 % of registered participants sent records, and the probability of a participant sending a record dropped off quickly over time after first registering. Among participants who contributed, ∼50 % sent 1 record, ∼30 % ≥3 and 4 % >10 records. Participants showed good capacity to identify mosquitoes and improve identification skills with app usage. The results will be valuable for anyone interested in evaluating citizen science, as participation and engagement are seldom quantitatively assessed. Our results are also useful for designing dissemination and education strategies in citizen science projects associated with arthropod vector monitoring.

Parasitic Characteristics of Ticks (Acari: Ixodidae) Collected from Water Deer (Hydropotes inermis argyropus) and Spatiotemporal Distribution Prediction within Host-Influenced Cultivated Areas.

Tick-borne diseases result from human-ixodid tick encounters, making it crucial to understand host-ixodid tick interactions and host-dependent distribution for epidemiology and prevention. This study examines water deer patterns and ixodid tick interactions in designated croplands of South Korea over two years, finding that the highest deer presence occurred in July and the lowest in May, during crop harvesting. Four tick species were identified, with Haemaphysalis longicornis being predominant (92.2%). Tick life stage analysis revealed peak nymphs and adults in July and larvae in October. Nymph abundance correlated positively with nearby water deer. MaxEnt biodiversity prediction results indicated wider water deer distribution in summer, reflecting their tendency to use multiple croplands. Areas with nymphs and adults aligned with predicted deer presence in summer, while larval areas aligned in autumn. Increased agroforestry expanded water deer habitats, enhancing tick dispersion. Prevention involved minimizing human-deer encounters by strategic land use in tick-prone areas. This comprehensive study provides insights into preventing severe fever with thrombocytopenia syndrome in agricultural workers, emphasizing the need for targeted interventions based on host behavior and tick life stages in different seasons.

Characterization of spatial lipidomic signatures in tick-bitten guinea pig skin as a model for host-vector-pathogen interaction profiling.

IMPORTANCE: Here, we demonstrate the adaptability of spatial "omics" methods to identify interphylum processes regulated at the vector-host interface of ticks during a mammalian blood meal. This approach enables a better understanding of complex bipartite or tripartite molecular interactions between hosts, arthropod vectors and transmitted pathogens, and contributes toward the development of spatially aware therapeutic target discovery and description.