This is the first study showing HEV to be present in Romanian pig herds and that the human population is exposed.

West Nile virus (WNV) is a zoonotic flavivirus whose transmission cycle in nature includes wild birds as amplifying hosts and ornithophilic mosquito vectors. Bridge vectors can transmit WNV to mammal species potentially causing West Nile Fever. Wild bird migration is a mode of WNV introduction into new areas. The Danube Delta Biosphere Reserve (DDBR) is a major stopover of wild birds migrating between Europe and Africa. The aim of this study was to investigate the presence of WNV in the DDBR during the 2016 transmission season in wild birds and mosquitoes. Blood from 68 wild birds (nine different species) trapped at four different locations was analyzed by competitive ELISA and Virus Neutralization Test (VNT), revealing positive results in 8/68 (11.8%) of the wild birds by ELISA of which six samples (three from juvenile birds) were confirmed seropositive by VNT. Mosquitoes (n = 6523, 5 genera) were trapped with CDC Mini Light traps at two locations and in one location resting mosquitoes were caught. The presence of WNV RNA was tested in 134 pools by reverse transcription quantitative PCR (RT-qPCR). None of the pools was positive for WNV-specific RNA. Based on the obtained results, WNV was circulating in the DDBR during 2016.

The purpose of this research was to improve the epidemiological data on HEV infection in the human population in Romania. The analysis targeted hospitalized subjects with acute hepatitis (n = 94) of unknown etiology from the Infectious Diseases Regional Hospital in Iasi. Moreover, patients without liver disease (n = 40) from a different county hospital located in Eastern Romania were included. The presence of HEV infection and first characterization of human HEV strains was determined using serological and molecular assays. The apparent HEV seroprevalence varied between 29.16% (95% CI, 16.31–42.03) and 32.5% (95% CI, 17.98–47.02) according to patient grouping. Molecular analysis enhanced the detection of two HEV isolates, that clustered in subtype HEV-3c, the most commonly identified subtype in Europe. Identification of acute hepatitis E cases, together with the first detection and molecular characterization of human HEV in Romania represent the originality attributes of the present study.

The impact of mosquito-borne diseases on human and veterinary health is being exacerbated by rapid environmental changes caused mainly by changing climatic patterns and globalization. To gain insight into mosquito-borne virus circulation from two counties in eastern and southeastern Romania, we have used a combination of sampling methods in natural, urban and peri-urban sites. The presence of 37 mosquito-borne viruses in 16,827 pooled mosquitoes was analyzed using a high-throughput microfluidic real-time PCR assay. West Nile virus (WNV) was detected in 10/365 pools of Culex pipiens (n = 8), Culex modestus (n = 1) and Aedes vexans (n = 1) from both studied counties. We also report the first molecular detection of Sindbis virus (SINV) RNA in the country in one pool of Culex modestus. WNV infection was confirmed by real-time RT-PCR (10/10) and virus isolation on Vero or C6/36 cells (four samples). For the SINV-positive pool, no cytopathic effectwas observed after infection of Vero or C6/36 cells, but no amplification was obtained in conventional SINV RT-PCR. Phylogenetic analysis of WNV partial NS5 sequences revealed that WNV lineage 2 of theCentral-Southeast European clade, has a wider circulation in Romania than previously known.

Ixodid ticks are competent vectors for multiple pathogens, several of which cause infections in human. The medical importance of tick-borne pathogens is well known, yet unanswered questions remain regarding the occurrence of pathogens such as Rickettsia spp., Anaplasma phagocytophilum, and "Candidatus Neoehrlichia mikurensis" in questing ticks in Romania. Our objectives were to identify three emerging tick-borne zoonotic pathogens in eastern Romania, to assess their prevalence, establish co-infection rates, and to compare infection levels of selected pathogens in questing ticks collected from one suburban area in the city of Iaşi and one forested area located in the Danube Delta Biosphere Reserve. We collected 490 questing nymphs or adult ticks (467 Ixodes ricinus, 4 Dermacentor reticulatus, and 19 Haemaphysalis punctata). We individually analyzed ticks for the presence of Rickettsia spp., A. phagocytophilum, and "C. N. mikurensis." Rickettsia spp. was detected in 9.4% of ticks from both sampling areas. Rickettsia spp. included R. helvetica (n = 17 I. ricinus ticks), R. monacensis (n = 28 I. ricinus ticks), and R. raoultii (n = 1 D. reticulatus). "C. N. mikurensis" had an infection rate of 4.9% while A. phagocytophilum was detected only in the forested area with a global prevalence of 1.2%. The overall prevalence of ticks infected with at least one pathogen was 15.5%, and 5.3% of infected ticks were tested positives for dual pathogen association. Our study documents the presence of pathogens in questing ticks in the urban recreational areas of Iaşi and forested areas located in the Danube Delta Biosphere Reserve. Worth mentioning, is the presence of "C. N. mikurensis" in ticks from eastern Romania, an agent just recently described in Romania, and the existence of co-infections in ticks at a similar prevalence to other European countries.