To date, the microbiome, as well as the virome of the Croatian populations of bats, was unknown. Here, we present the results of the first viral metagenomic analysis of guano, feces and saliva (oral swabs) of seven bat species (Myotis myotis, Miniopterus schreibersii, Rhinolophus ferrumequinum, Eptesicus serotinus, Myotis blythii, Myotis nattereri and Myotis emarginatus) conducted in Mediterranean and continental Croatia. Viral nucleic acids were extracted from sample pools, and analyzed using Illumina sequencing. The presence of 63 different viral families representing all seven Baltimore groups were confirmed, most commonly insect viruses likely reflecting the diet of insectivorous bats. Virome compositions of our samples were largely impacted by the sample type: invertebrate-infecting viruses were most frequently found in feces, bacterial viruses in guano, whereas vertebrate-infecting viruses were most common in swabs. Most vertebrate-infecting virus sequences were assigned to retroviruses, parvoviruses, iridoviruses, and poxviruses. We further report the complete genome sequence of a novel adeno-associated virus, densovirus and a near complete length genome sequence of a novel iflavirus. Additionally, one of the most interesting findings in this study was the difference in viromes between two contrasting habitats, the continental and Mediterranean Croatia.

BackgroundRabies is the only known zoonotic disease of bat origin in Europe. The disease is caused by species belonging to the genus Lyssavirus. Five Lyssavirus species, i.e., European bat lyssavirus (EBLV)-1, EBLV-2, Bokeloh bat lyssavirus, Lleida bat lyssavirus, and West Caucasian bat virus, have been identified in European bats. More recently, a proposed sixth species, Kotalahti bat lyssavirus, was detected. Thus, in this study, active surveillance was initiated in order to obtain insights into the prevalence of lyssaviruses in Croatian bat populations and to improve our understanding of the public health threat of infected bats.ResultsIn total, 455 bats were caught throughout Continental and Mediterranean Croatia. Antibodies were found in 20 of 350 bats (5.71%, 95% confidence interval 3.73–8.66). The majority of seropositive bats were found in Trbušnjak cave (Continental Croatia, Eastern part), and most seropositive bats belonged to Myotis myotis (13/20). All oropharyngeal swabs were negative for the presence of Lyssavirus.ConclusionsThe presence of lyssaviruses in bat populations was confirmed for the first time in Croatia and Southeastern Europe. The results of this study suggest the need for further comprehensive analyses of lyssaviruses in bats in this part of Europe.

A.: DNA barcoding of the family Phryganeidae (Insecta, Trichoptera) in Croatia with particular reference to phylogeny, distribution and conservation biology. Nat. Croat., Vol. 28, No. 2., 305-323, 2019, Zagreb.In Europe the Phryganeidae family comprises 7 genera and 20 species. In Croatia, 6 species have been recorded from this family, and four of them recorded and collected in the last five years have been DNA barcoded. In terms of faunistic research, most interesting is the record of Trichostegia minor Curtis, 1834 from the region of the Gacka River in Lika, in upland Croatia. Up to now four species have been included in the BOLD system with 6 DNA-barcoded specimens. From the Gacka River area, 11 specimens of 8 species, including T. minor, have been barcoded and included into the BOLD system. Phylogenetic research based on DNA barcode data has shown some interesting results. Specimen of T. minor collected in the area of the Gacka River is clustered with a specimen of this species from Austria into a separate subgroup, which shows minor molecular specifics. Phylogenetic analysis has confirmed the justification of the status of a subspecies from Mongolia, Phryganea grandis rotundata Ulmer, 1905.Future Trichoptera fauna research in Croatia will continue investigations of the family Phryganeidae in various parts of the country for the purpose of ascertaining the detailed distribution of all the species recorded, to enable finding some other previously missing species, as well as collecting and DNA barcoding of Hagenella clathrata (Kolenati, 1848) and Oligostomis reticulata (Linnaeus, 1761). With respect to these species, the data will also be useful for the area of conservation biology, necessary for their protection and for the protection of the sites in which they occur, for it is clear that these are very rare species in Croatia, T. minor also belonging to this group: 50% of the species from this family recorded in Croatia are rare and very rare species. Kučinić, M. et al.: DNA barcoding of the family Phryganeidae in Croatia with particular reference to ...306 Kučinić, M., Ćukušić, A., Cerjanec, D., Podnar, M., Plantak, M., Žalac, S., Ćuk, R., Vučković, I., Ibrahimi, H. & Delić, A.: DNA barkodiranje porodice Phryganeidae (Insecta, Trichoptera) u Hrvatskoj, s posebnim osvrtom na filogeniju, rasprostranjenost i konzervacijsku biologiju. Nat. Croat., Vol. 28, No. 2., 305-323, 2019, Zagreb. Porodica Phryganeidae broji u Europi 7 rodova i 20 vrsta. Iz ove porodice na području Hrvatske utvrđeno je 6 vrsta, od kojih su DNA barkodirane četiri vrste, zabilježene i prikupljene u posljednjih pet godina. Faunistički najzanimljiviji je nalaz vrste Trichostegia minor Curtis, 1834 s područja rijeke Gacke (regija Lika), gorska Hrvatska. U BOLD bazu upisane su četiri vrste sa šest DNA barkodiranih primjeraka. S područja rijeke Gacke je DNA barkodirano i upisano u BOLD bazu 11 primjeraka s osam vrsta, uključujući i vrstu T. minor. Filogenetska istraživanja temeljena na analizi DNA barkod podataka pokazala su zanimljive rezultate...

Caddisfly faunistic and ecological features were studied at 14 sampling sites along the catchment area of the Dobra River. Adults were collected monthly, over a period of 13 months (from early November 2009 until the end of November 2010), using ultraviolet (UV) light trap and entomological net. Physico-chemical parameters of water and air were also measured at all sampling sites. The study encompassed several segments of caddisfly fauna: the composition, new records, the distribution of all recorded species and seasonal dynamics. A total number of 77 species were recorded, out of which 15 species and four genera were new records for Croatian caddisfly fauna. The CCA analysis showed that the water temperature and the altitude had the highest impact on distribution of caddisfly fauna in this area. The results of this study contribute to the increase of faunistic and ecological knowledge of the Croatian caddisflies.

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