First Record of West Nile Virus Specific Seroconversion in Dogs From Eastern Romania

West Nile virus (WNV) infection causes diseases that vary in intensity from asymptomatic to fatal encephalitis in humans and animals. The etiological agent is a Flavivirus belonging to Japanesse Encephalitis complex. This zoonotic virus is maintained in nature in an enzootic cycle between mosquitoes and birds as amplifying hosts. Except birds, a wide variety of animals develop seroconversion caused by infection, and previous studies indicated that dogs may be considered as sentinel for WNV. The importance of studying this virus is caused by its zoonotic character, limited possibilities of active surveillance and real-time alertness of the infection’s presence in the human and animal populations. The aim of the study was to demonstrate and estimate the seroconversion in dogs within an area with confirmed cases in humans and animals. A total number of 76 serum samples were evaluated for the presence of specific anti WNV antibodies using a commercial competitive ELISA kit, for the detection of anti-pr-E antibodies (ID Screen® West Nile Competition Multi-species). The results indicated specific antibodies in 32 (42.1%) out of 76 samples tested. Positive results were recorded in all four counties from which samples were collected. Our preliminary results proved the circulation of WNV in dogs in areas where the infection was previously reported in animals and humans

Species diversity, host preference and arbovirus detection of Culicoides (Diptera: Ceratopogonidae) in south-eastern Serbia

BackgroundCulicoides (Diptera: Ceratopogonidae) is a genus of small biting midges (also known as no-see ums) that currently includes 1368 described species. They are proven or suspected vectors for important pathogens affecting animals such as bluetongue virus (BTV) and Schmallenberg virus (SBV). Currently little information is available on the species of Culicoides present in Serbia. Thus, the aim of this study was to examine species diversity, host preference and the presence of BTV and SBV RNA in Culicoides from the Stara Planina Nature Park in south-eastern Serbia.ResultsIn total 19,887 individual Culicoides were collected during three nights of trapping at two farm sites and pooled into six groups (Obsoletus group, Pulicaris group, Others group and further each group according to the blood-feeding status to freshly engorged and non-engorged). Species identification was done on subsamples of 592 individual Culicoides specimens by morphological and molecular methods (MALDI-TOF mass spectrometry and PCR/sequencing). At least 22 Culicoides species were detected. Four animal species (cow, sheep, goat and common blackbird) as well as humans were identified as hosts of Culicoides biting midges. The screening of 8291 Culicoides specimens in 99 pools for the presence of BTV and SBV RNA by reverse-transcription quantitative PCR were negative.ConclusionsThe biodiversity of Culicoides species in the natural reserve Stara Planina was high with at least 22 species present. The presence of C. imicola Kieffer was not recorded in this area. Culicoides showed opportunistic feeding behaviour as determined by host preference. The absence of SBV and BTV viral RNA correlates with the absence of clinical disease in the field during the time of sampling. These data are the direct outcome of a training programme within the Institutional Partnership Project AMSAR: Arbovirus monitoring, research and surveillance-capacity building on mosquitoes and biting midges funded by the programme SCOPES of the Swiss National Science Foundation

Preliminary Data Regarding the Nontuberculous Mycobacteria Isolation from Clinical Specimens, from Human PatienTS, in Iași County

The nontuberculous mycobacteria (NTM) represents a large group ofubiquitous environmental mycobacterial species, with more than 160 members that can pose a threat for human health, especially for the immunocompromised hosts. The aim of the current study was the collection of data regarding the frequency of NTM isolation from human clinical specimens, thus providing preliminary data regarding the infection prevalence in Iași County, Romania.Between May 2015 and March 2017 epidemiological data regarding NTM isolatesin the Bacteriology Laboratory at the Clinical Hospital of Pneumology in Iasi were analyzed. A total of 63 mycobacteria strains were isolated and identified as nontuberculous (by smear microscopy, culture on solid and liquid media and immunochromatographic assay). Out of the total number of samples that were submitted in the laboratory for the diagnosis of tuberculosis, during the studied period, 0.25% of them were identified as NTM. The prevalence for 2015 was estimated at 0.18%, and 0.34% for 2016. Given the challenging diagnostic procedures and the absence of notifiable status of NTM in most European Union countries, the epidemiological situation is broadlyunknown. This emphases the need of regular recording and reporting of NTM isolation frequency for awareness of the risk of associated mycobacterial infections

First Detection of Hepatitis E Virus (<i>Rocahepevirus ratti</i> Genotype C1) in Synanthropic Norway Rats (<i>Rattus norvegicus</i>) in Romania

Hepatitis E virus (HEV) is an emerging zoonotic pathogen with different viral genera and species reported in a wide range of animals. Rodents, particularly rats, carry the specific genus rat HEV (Rocahepevirus genus, genotype C1) and are exposed occasionally to HEV-3 (Paslahepevirus genus, genotype 3), a zoonotic genotype identified in humans and widely distributed in domestic and feral pigs. In this study, the presence of HEV was investigated in synanthropic Norway rats from Eastern Romania, in areas where the presence of HEV-3 was previously reported in pigs, wild boars and humans. Using methods capable of detecting different HEV species, the presence of HEV RNA was investigated in 69 liver samples collected from 52 rats and other animal species. Nine rat liver samples were identified as being positive for rat HEV RNA (17.3%). High sequence identity (85–89% nt) was found with other European Rocahepevirus. All samples tested from other animal species, within the same environment, were negative for HEV. This is the first study to demonstrate the presence of HEV in rats from Romania. Since rat HEV has been reported to cause zoonotic infections in humans, this finding supports the need to extend the diagnosis of Rocahepevirus in humans with suspicion of hepatitis

Detection of West Nile Virus Lineage 2 in Eastern Romania and First Identification of Sindbis Virus RNA in Mosquitoes Analyzed using High-Throughput Microfluidic Real-Time PCR

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