Molecular analysis of blood-associated pathogens in European wildcats (Felis silvestris silvestris) from Germany.

European wildcats (Felis silvestris silvestris) have not been investigated in large numbers for blood-associated pathogens in Germany, because wildcats, being a protected species, may not be hunted, and the collection of samples is therefore difficult. Thus, spleen tissue and whole blood from 96 wildcats from Germany found as roadkill or dead from other causes in the years 1998-2020 were examined for the prevalence of blood associated pathogens using molecular genetic tools. PCR was used to screen for haemotrophic Mycoplasma spp., Hepatozoon spp., Cytauxzoon spp., Bartonella spp., Filarioidea, Anaplasmataceae, and Rickettsiales, and positive samples were subsequently sequenced. Phylogenetic analyses were performed for Mycoplasma spp. and Hepatozoon spp. by calculating phylogenetic trees and DNA haplotype networks. The following pathogens were found: Candidatus Mycoplasma haematominutum (7/96), Mycoplasma ovis (1/96), Hepatozoon silvestris (34/96), Hepatozoon felis (6/96), Cytauxzoon europaeus (45/96), and Bartonella spp. (3/96). This study elucidates the prevalence of blood-associated pathogens in wildcats from Germany.

First Nationwide Monitoring Program for the Detection of Potentially Invasive Mosquito Species in Austria.

In Austria, only fragmented information on the occurrence of alien and potentially invasive mosquito species exists. The aim of this study is a nationwide overview on the situation of those mosquitoes in Austria. Using a nationwide uniform protocol for the first time, mosquito eggs were sampled with ovitraps at 45 locations in Austria at weekly intervals from May to October 2020. The sampled eggs were counted and the species were identified by genetic analysis. The Asian tiger mosquito Aedes albopictus was found at two sites, once in Tyrol, where this species has been reported before, and for the first time in the province of Lower Austria, at a motorway rest stop. The Asian bush mosquito Aedes japonicus was widespread in Austria. It was found in all provinces and was the most abundant species in the ovitraps by far. Aedes japonicus was more abundant in the South than in the North and more eggs were found in habitats with artificial surfaces than in (semi-) natural areas. Further, the number of Ae. japonicus eggs increased with higher ambient temperature and decreased with higher wind speed. The results of this study will contribute to a better estimation of the risk of mosquito-borne disease in Austria and will be a useful baseline for a future documentation of changes in the distribution of those species.

Emergence of Parafilaria bovicola in Austria.

Veterinarians reported cases of cutaneous bleeding in cattle in Austria in the spring and summer of 2020. It was our goal to confirm the tentative diagnosis of parafilariosis by identifying Parafilaria bovicola in exudate samples using molecular methods for the first time in Austria. We asked veterinarians in the field to collect exudate from typical lesions on cattle. We performed polymerase chain reactions (PCRs) and sequenced a 674-bp section of the mitochondrial cytochrome oxidase subunit I in all positive samples. Overall, in 57 of 86 samples, P. bovicola was confirmed by PCR in cattle from Lower Austria, Upper Austria, Styria, Salzburg, Carinthia, and Tyrol. Sequencing detected four different haplotypes or genotypes, respectively, indicating multiple routes of introduction. We conclude that parafilariosis has spread in Austria and we expect that the number of reports of clinical signs and losses due to carcass damage will increase in the future.

Monitoring of alien mosquitoes in Western Austria (Tyrol, Austria, 2018).

Mosquitoes are of major importance to human and animal health due to their ability to transmit various pathogens. In Europe the role of mosquitoes in public health has increased with the introduction of alien Aedes mosquitoes such as the Asian tiger mosquito, Aedes albopictus; the Asian bush mosquito, Ae. japonicus; and Ae. koreicus. In Austria, Ae. japonicus has established populations in various regions of the country. Aedes albopictus is not known to overwinter in Austria, although isolated findings of eggs and adult female mosquitoes have been previously reported, especially in Tyrol. Aedes koreicus had not so far been found in Austria. Within the framework of an alien mosquito surveillance program in the Austrian province of Tyrol, ovitraps were set up weekly from May to October, 2018, at 67 sites- 17 in East Tyrol and 50 in North Tyrol. Sampling was performed at highways and at urban and rural areas. DNA obtained from mosquito eggs was barcoded using molecular techniques and sequences were analysed to species level. Eggs of alien Aedes species were found at 18 out of 67 sites (27%). Both Ae. albopictus and Ae. japonicus were documented at highways and urban areas in both East and North Tyrol. Aedes koreicus was found in East Tyrol. During this mosquito surveillance program, eggs of Ae. albopictus, Ae. japonicus, and Ae. koreicus were documented in the Austrian province of Tyrol. These findings not only show highways to be points of entry, but also point to possible establishment processes of Ae. japonicus in Tyrol. Moreover, Ae. koreicus was documented in Austria for the first time.

Monitoring of alien mosquitoes of the genus Aedes (Diptera: Culicidae) in Austria.

Systematic, continuous mosquito surveillance is considered the most reliable tool to predict the spread and establishment of alien mosquito species such as the Asian tiger mosquito (Aedes albopictus), Japanese bush mosquito (Aedes japonicus), and the transmission risk of mosquito-borne arboviruses to humans. Only single individuals of Ae. albopictus have been found in Austria so far. However, it is likely that the species will be able to establish populations in the future due to global trade and traffic as well as increasing temperatures in the course of global climate change. In summer 2017, a project surveilling the oviposition of newly introduced Aedes mosquitoes, using ovitraps, was set up by means of citizen scientists and researchers and was performed in six federal provinces of Austria-Tyrol, Carinthia, Vienna, Lower Austria, Styria, and Burgenland. Eggs of Ae. albopictus were identified in Tyrol during the months August and September, while Ae. japonicus was found in Lower Austria, Styria, and Burgenland. In Vienna and Carinthia, all ovitraps were negative for Aedes eggs; however, Ae. japonicus was found for the first time in Vienna in July 2017 during routine sampling of adult mosquitoes. With this project, we demonstrated the benefits of citizen scientists for ovitrap-based mosquito surveillance. The finding of Ae. albopictus eggs in Northern Tyrol is not yet a proof of the establishment of a self-sustaining population, although it indicates the ongoing introduction of this species along main traffic routes from Italy, where this mosquito is well established. The risk of establishment of the tiger mosquito in the Lower Inn Valley is therefore a given and informing the public about preventive measures to hinder and delay this development is highly recommended.