The European subtype of tick-borne encephalitis virus (TBEV-Eur; species Orthoflavivirus encephalitidis, family Flaviviridae) was the only tick-borne flavivirus present in central Europe known to cause neurologic disease in humans and several animal species. Here, we report a tick-borne flavivirus isolated from Alpine chamois (Rupicapra rupicapra rupicapra) with encephalitis and attached ticks, present over a wide area in the Alps. Cases were detected in 2017 in Salzburg, Austria, and 2023 in Lombardy and Piedmont, Italy. The virus strains exhibit 94.8–97.3% nucleotide identities to each other and are more closely related to Louping ill viruses (LIV; Orthoflavivirus loupingi; 90–92% identities) than to TBEV-Eur (less than 88%). The chamois-derived virus strains, tentatively termed “Alpine chamois encephalitis virus”, form a well-supported independent genetic clade with Spanish goat encephalitis virus, clearly separated from other LIV. This supports its designation as a new virus subtype with the proposed shared taxonomic name “Spanish goat and Alpine chamois encephalitis virus subtype” within the species Orthoflavivirus loupingi. The zoonotic potential of this newly identified virus subtype as well as its host range in other animal species including farm animals needs to be further investigated.

The cultivation of Chlamydia gallinacea, a recently identified species, is challenging due to the lack of an optimized protocol. In this study, several infection protocols were tested, including different cell lines, incubation temperatures, centrifugation methods and culture media. However, none were successful in field samples. The only exception was a chance co-culture with Trichosporon asahii, a microorganism commonly found in the chicken gut. This suggests that current in vitro methods may not be optimized for this species and that host-associated microorganisms may influence the in vivo growth of C. gallinacea, which is typically found in the chicken gut. These findings raise new questions and highlight the need for further investigation of microbial interactions within the host, particularly to understand their role in the proliferation of chlamydial species.

Background/Objectives: The detection of anti-Coxiella antibodies using serological methods is essential for identifying exposed ruminants and preventing this important zoonotic disease in livestock. In recent years, numerous attempts have been made to increase diagnostic performance as well as simplify the production of serological assays. Commercially available tests often use whole-cell antigens, which can decrease specificity and require high-level biosafety facilities for manufacturing. The aim of this work was to produce three Coxiella burnetii (C. burnetii) antigens in recombinant form and assess them for the detection of anti-Coxiella antibodies in ruminants. Methods: Three recombinant C. burnetii antigens (Com-1, MceB, AdaA) were selected among immunodominant antigens and produced in a heterologous system (Escherichia coli). Following purification, the proteins were utilized to coat ELISA plates and evaluated for seroreactivity against sera from both negative and positive cattle. Results: Com-1 demonstrated the greatest agreement with the commercial test, albeit moderate. MceB exhibited nonspecific reactivity against a large number of sera, while the AdaA showed reactivity against only a few positive sera. Conclusions: Our findings are consistent with previous research, indicating that utilizing a single antigen to identify exposed animals is unfeasible with current knowledge, most likely due to the complex immunological response following C. burnetii infection in cattle. Consequently, it is critical to continue testing and identifying immunoreactive antigens in order to further investigate them and, potentially, select the most appropriate.

Since its introduction to Europe in 2004, West Nile Virus Lineage 2 (WNV-2) has become endemic, with Italy reporting the highest number of cases each season. In 2022, WNV infections in Italy exceeded those recorded during the major 2018 outbreak—the largest ever reported in Europe. This study investigates the genomic epidemiology of WNV during the 2022 and 2023 transmission seasons.

Tick-borne diseases are emerging worldwide due to the expanding distribution of tick vectors and represent an increasing public health concern. This study investigated the occurrence of major tick-borne pathogens in ticks collected from wild animals in Valle d’Aosta Region (Northwestern Italy) between 2017 and 2022. Tick pools were screened for Anaplasma spp., Borrelia burgdorferi s.l., Coxiella burnetii, Rickettsia spp., and Tick-Borne Encephalitis virus (TBEV). Most collected ticks were morphologically identified as Ixodes ricinus. Overall, 15.2% of the 158 analyzed pools tested positive for at least one pathogen. Rickettsia spp. was the most frequently detected pathogen (10.7%), with multiple species identified, including R. helvetica, R. monacensis, R. slovaca, and R. raoultii. Anaplasma phagocytophilum was detected in ticks collected from wild ungulates, whereas Borrelia burgdorferi s.l. was only occasionally identified. No samples tested positive for TBEV or C. burnetii. Rickettsia spp. positivity was higher in ticks collected from ungulates and in I. ricinus, although these differences were not statistically significant. These findings provide new insights into tick ecology in Alpine ecosystems, highlighting the circulation of potentially zoonotic pathogens and the altitudinal presence of ticks in the Alps up to 1800–2300 m a.s.l. The study contributes valuable epidemiological data and supports the need for continued surveillance within a One Health framework.