West Nile virus infections in Romania after the 2010 outbreak - a retrospective study in human and animal population

In this study the authors have made a retrospective analysis of West Nile virus infection in human and animals after 2010. Few studies have dealt with this arbovirosis in the last years and there is no analysis to correlate the evolution in animals and humans. To date it is considered that the largest outbreak of West Nile virus (WNV) encephalitis from Europe was registered in 1996 in Romania. Cases of encephalitis with WNV have been reported in our country before first in 1955 in Transylvania, followed by an outbreak in 1964 in Banat. The outbreak from 1996, ended with over 390 confirmed cases of encephalitis in humans the majority being registered in Bucharest and in the southern area of Romania. The epidemiological situation, after the implementation of a surveillance system for WNV in humans until 2010, was characterized by sporadic cases, WNV being reported yearly, except 2002. The infections were reported mostly in the south and south eastern area of the country. The age distribution in human cases recorded a majority in the group age over 65 years due to the senescence of the immune system and the associated diseases. The gender ratio was dominant for men, considering the occupational factor. In 2010 the second large outbreak of WNV was recorded and 47 confirmed cases of WNV infection were registered. After 2010, WNV encephalitis was recorded yearly and the area of detection was confined to eastern area and slowly to the to the whole country. Human cases were registered yearly in areas where the virus was detected in animals by serological screening. Considering the analyzed data, we can state that the main feature of WNV infection is the continuous virus spreading over the territory both in humans and animals. This can be determined by the climatic changes, that allow the surviving and multiplication of the competent vectors in new areas and maybe of the changes in the migratory routes of the birds- main amplifying hosts. Unfortunately, no statistical correlation between human and animal cases could be made. We can state that WNV is continuing to represent a threat for public health and more in depth research has to be made in order to characterize the evolution of the infection in our country and the causes of its endemisation

Detection of BHV-1 antibodies in bovine raised in rural households from Iași County

Infectious bovine rhinotracheitis / infectious pustular vulvovaginitis (IBR/IPV) is a significant disease among domestic and wild cattle. IBR/IPV is caused by Bovine Herpesvirus-1 (BHV-1) that is capable of attacking many different tissues in the body leading to a variety of clinical diseases. The virus can persist in clinically recovered animals for years, remaining inactive until the animal is placed under stress. Investigations were undertaken during 2015-2016 on bovine raised in rural households in Miroslovești and Ciohorăni commune from Iași County. Individual serum samples from 305 cattle were tested for detection of BHV-1 antibodies using an ELISA commercial kit (IDEXX IBR Ab). The results showed that overall seroprevalence of infectious bovine rhinotracheitis (IBR) was 63.6% in tested cattle. These data underline the fact that in household reared cattle are several animals with BHV-1 lifelong latent infection

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

Diversity of nontuberculous mycobacteria isolated from animal samples

Limited data is available regarding the epidemiology of the nontuberculous mycobacteria (NTM) in Romania, in both human and veterinary medicine. The objective of the present study was the identification and characterization of the Mycobacterium avium complex species isolated from backyard poultry and ruminant samples. The molecular identification and genotyping was performed in ANSES, Laboratory for Animal Health, Maisons-Alfort, France, by real time PCR, with a wide range of targets: hsp65, IS6110, IS1081, IS1245, IS901 and MIRU-VNTR. M. avium avium and M. avium paratuberculosis were the two species identified. Five different profiles were obtained through genotyping, four of which had corresponding INMV (INRA Nouzilly MIRU-VNTR) profiles: INMV 2, INMV 67, INMV 99 and INMV 100. The technique differentiated between the M. avium paratuberculosis isolated from sheep and cattle and expressed a high discriminatory power, proving to be extremely useful for assessing the genetic diversity of the tested animal origin samples and providing comparable information on the general structure and main pathogens belonging to MAC

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