https://www.veterinariaitaliana.izs.it/index.php/VetIt <p>A quarterly peer-reviewed journal devoted to veterinary public health and other aspects of veterinary science and medicine, Veterinaria Italiana is published by the Istituto Zooprofilattico Sperimentale dell’Abruzzo e del Molise ‘G. Caporale’ (Istituto G. Caporale) in Teramo, Italy.</p> Istituto Zooprofilattico Sperimentale dell'Abruzzo e del Molise G. Caporale en-US Veterinaria Italiana 0505-401X https://www.veterinariaitaliana.izs.it/index.php/VetIt/article/view/3946 <p>Bluetongue virus (BTV) and epizootic haemorrhagic disease virus (EHDV) are Culicoides-borne orbiviruses affecting domestic and wild ruminants. Information on their circulation in Namibian wildlife is limited. This study investigated the molecular detection and serotype distribution of BTV and EHDV in wild ruminants from a commercial game farm in the Khomas Region, Namibia, where wildlife and livestock coexist.</p> <p>Between June and September 2019, spleen samples from 62 clinically healthy animals (kudu, oryx, and red hartebeest) were analysed by real-time RT-PCR using pan-BTV and pan-EHDV assays, followed by serotype-specific tests for selected BTV types. Two animals (3.23%) tested positive for EHDV. BTV RNA was detected in 24/62 animals (38.71%), with Ct values ranging from 28.3 to 38.4. BTV-3 and BTV-4 were the most frequently identified serotypes, while one sample was positive for BTV-1; six BTV-positive samples remained untyped. High Ct values and low RNA loads likely limited sequencing success.</p> <p>Although restricted to a single farm and a limited serotype panel, this study provides preliminary molecular evidence of BTV and EHDV circulation in Namibian wild ruminants, highlighting the need for broader epidemiological investigations at the wildlife–livestock interface.</p> Umberto Molini Gloria Plebani Maria Yvonne Hemberger Nicandro Rodi Mariassunta Iannetta Ottavio Portanti Alessio Lorusso Juliet Kabajani Copyright (c) 2026 Umberto Molini, Gloria Plebani, Maria Yvonne Hemberger, Nicandro Rodi, Mariassunta Iannetta, Ottavio Portanti, Alessio Lorusso, Juliet Kabajani http://127.0.0.1/foo.html 2026-03-04 2026-03-04 62 2 10.12834/VetIt.3946.39700.1 https://www.veterinariaitaliana.izs.it/index.php/VetIt/article/view/3908 <p>Pathogen-agnostic diagnostics are crucial for the early detection of emerging viruses. Shotgun metagenomic sequencing enables unbiased detection of viral genomes but is frequently constrained by the abundance of host and microbial ribosomal RNA (rRNA), which reduces sensitivity and increases sequencing costs. CRISPR-Cas9–based rRNA depletion has emerged as an alternative to enzymatic methods; however, its performance for the characterization of zoonotic viruses across diverse animal hosts and tissues remains underexplored. We compared CRISPR-Cas9 (Jumpcode CRISPRclean™ Plus) and RNase H–based enzymatic depletion (Ribo-Zero Plus, Illumina) using 12 samples positive for rabies lyssavirus, influenza A virus, West Nile virus or norovirus, from multiple host species and tissues, including both high-quality and degraded RNA. CRISPR-Cas9 efficiently reduced rRNA content (14.5%) but recovered fewer viral reads than Ribo-Zero, which achieved up to 60.7× enrichment. Both methods produced complete viral consensus genomes when RNA quality and viral load were sufficient. However, based on the data generated here, enzymatic depletion currently remains more efficient and cost-effective for viral metagenomics. Further optimization of CRISPR-Cas9 workflows could enhance its utility for viral surveillance and diagnostics.</p> Gianpiero Zamperin Elisa Palumbo Martina Castellan Sabrina Marciano Alice Fusaro Isabella Monne Copyright (c) 2026 Gianpiero Zamperin, Elisa Palumbo, Martina Castellan, Sabrina Marciano, Alice Fusaro, Isabella Monne http://127.0.0.1/foo.html 2026-03-04 2026-03-04 62 2 10.12834/VetIt.3908.38985.2