Southeastern Romania Research Articles

Obstructive Sleep Apnea after COVID-19: An Observational Study.

The risk factors of hospitalized COVID-19 and obstructive sleep apnea (OSA) overlap. The aim of this study is to evaluate the prevalence and associated factors of post-COVID-19 OSA in hospitalized adult patients from southeastern Romania. A follow-up study was conducted on patients hospitalized for COVID-19 at the Pneumology Hospital in Galati, Romania, between 2021 and 2022. OSA was evaluated using the Epworth and STOP-BANG questionnaires and nocturnal polygraphy monitoring. Out of 331 patients, 257 were evaluated for sleep apnea in the 12th week. The prevalence of severe OSA was 57.97%. Significant associations were found with male gender, an age over 60, obesity, and cardiovascular co-morbidities. Non-invasive ventilatory therapy (NIV) and a hygienic-dietary regimen were recommended based on severity following a control visit after a month. Developing strategies for diagnosing and monitoring sleep disorders, including home sleep apnea tests and patient education, are the next directions for post-COVID-19 management.

Age estimation in tortoises: an evaluation of methods for the spur-thighed tortoise (Testudo graeca)

Abstract Age and growth-related parameters provide a better understanding of life-history adaptations and trade-offs and can be studied in the wild through capture-mark-recapture (CMR) or sclerochronology (i.e., skeletochronology and counting growth marks on hard structures). In chelonians, skeletochronology is limited to carcasses, CMR is limited by time and funding constraints, but counting shell growth annuli (SGA count) is a cheaper and faster alternative. A long-term CMR study in Testudo graeca from south-eastern Romania allowed us to compare the age estimated by the two sclerochronology methods using carcasses, and calibrate the results with data obtained from recaptured tortoises. We found that the accuracy of SGA counts significantly decreased with time (i.e., years between captures and older individuals). Skeletochronology showed the best results with long bones (i.e., ulna, radius, femur), but overall underestimated the age when compared to SGA counts; we obtained comparable results from these two methods in tortoises up to 17 years old. The oldest age estimated by skeletochronology alone was 28 years, 30 years by SGA counts, and 40 years by SGA counts when calibrated by CMR. The growth models using age estimations from both skeletochronology and SGA counts were similar, highlighting their usefulness in monitoring growth trajectories. Our study showed that while age estimation in tortoises is challenging, a combination of methods can improve its accuracy.

New species of gall wasps (Hymenoptera: Cynipidae: Aulacideini and Phanacidini) inducing galls on plants of the genera Sonchus and Crepis (Asteraceae) in Romania (Dobrogea Province) and notes on their parasitoids

Two new species of herb gall wasps (Hym., Cynipidae) developing on plants in the genera Sonchus L. and Crepis L. (Asteraceae) are described from Southeastern Romania (Dobrogea Province): Aulacidea andriescui sp. n. gall inducer on Sonchus palustris L. and Phanacis dobrogicus sp. n. gall maker on Crepis foetida subsp. rhoeadifolia (M. Bieb.) Čelak. In addition to the diagnosis, identification, morphological description, life-cycle, geographical distribution, photos of gall and host plant, and SEM images, data on the associated fauna obtained from the galls of the two new species are also presented.

Monosensitisation to Salsola kali pollen pectin methylesterase in a patient from Southeastern Romania

Monosensitisation to Salsola kali pollen pectin methylesterase in a patient from Southeastern Romania

Assessment of Systemic Condition and Smoking Impact Over Incidence of Apical Periodontitis.

This study aimed to assess the prevalence of apical periodontitis in a subset of the population of south-eastern Romania and to analyze the overall health status of the group of patients with apical periodontitis. The medical and dental history, including age, gender, background, presence of smoking, level of education, the total number of teeth present and with apical periodontitis, and the general health status were recorded from a total of 200 patients. The periapical status was analyzed using an orthopantomogram and periapical radiographs of teeth that were diagnosed with periapical lesions by the same dental professional. The periapical status was classified according to the periapical index (PAI), withapical periodontitis being present if the PAI score ≥3. The majority of patients were female (58.5%), with secondary or higher education from urban areas and the mean number of teeth with apical periodontitis was 2.29 ± 1.26, with a median of 2 teeth. A total of 17.1% of patients were smokers, these patients had two more teeth with periapical pathology, and 16% of all patients had general diseases, the most common of which was cardiovascular disease (8.2%). Compared with those without the disease, these patients had a higher number of teeth with apical periodontitis (median = 2.5, IQR = 2-4 vs. median = 2, IQR = 1-3). As a result, this scientific research suggests an association between smoking, cardiovascular disease, and gastritis with apical periodontitis, but no association could be demonstrated between apical periodontitis and other systemic diseases.

Assessing Benzene and TVOC Pollution and the Carcinogenic and Noncarcinogenic Risks to Workers in an Industrial Plant in Southeastern Romania.

The article aims to analyze the pollution with Volatile Organic Compounds (VOC) emitted from the biggest refinery in Romania, using the daily and monthly series registered for two years in two sites on the industrial platform, and the carcinogenic and noncarcinogenic risks for workers at the industrial plant. Since the values of the basic statistics (minimum, maximum, and average) and outliers indicate that most recorded values exceed the maximum admissible limits established by law, the Peaks Over Threshold (POT) method was utilized to model the maximum values of the series and determine the return levels for benzene and total VOC (TVOC). Given the high values obtained for relatively short return periods, indicating potential danger for the workers, we assessed the noncarcinogenic and carcinogenic risks to benzene and TVOC exposure by computing the hazard index (HI) and lifetime cancer risk (LCR). The results indicate that 43.75% of the HI values are above 1, indicating a relatively high noncarcinogenic risk for different categories of workers. LRC indicates a high LRC for 93.75% of the workers in all considered categories exposed to TVOC.

Groundwater Exploration in Carbonate Reservoirs Using Borehole Investigations: A Case Study from South Dobrogea, Romania

The Late Jurassic–Early Cretaceous (J3–K1) transboundary aquifer is the most important groundwater body in southern–southeastern Romania, shared with Bulgaria and hosted in karstic–fractured carbonates. We conducted an integrated evaluation of this aquifer by analyzing three 700 m deep groundwater exploration–exploitation boreholes, which intercepted it in the Cernavodă area (South Dobrogea region). The evaluation was based on geophysical wireline logging, drilling information, and borehole production tests. A K-means clustering of the logging data was performed for lithology typing, formation boundaries identification, and the delineation of probable producing intervals associated with secondary porosity development. Petrophysical interpretation was carried out via depth-constrained (zonal) inversion, using multimineral models, the estimated formation boundaries, and variable uncertainties for the main input logs. The optimal interpretation models were correlated with borehole testing results to gain insight into the hydrogeological properties of the aquifer complex. The fractured–vuggy interval with the highest water-producing potential was identified in the lower section of the J3-age Rasova Formation (639–700 m depth), comprising mainly undolomitized limestones. A southeast-to-northwest trend of increasing productivity of the boreholes, correlated with an increasing lateral dolomitization intensity within the Rasova Formation, suggests a highly heterogeneous character of the aquifer. The differences in productivity are due not only to local porosity variations but also to various degrees of pore space connectivity that are related to the amount of fracturing or karstification. The novel findings of this study have important practical implications for the optimal placement, design, and drilling program of future groundwater exploitation boreholes in the Cernavodă area and neighboring sectors.

SEROLOGICAL SCREENING FOR Neospora caninum INFECTION IN A DAIRY CATTLE FARM WITH REPRODUCTIVE DISORDERS, IN SOUTH-EASTERN ROMANIA

Neospora caninum is an apicomplexan protozoa parasite that causes reproductive disorders in cattle worldwide. Despite the high seroprevalence reported in Romania, there are limited data on N. caninum infection in dairy cattle, and no studies have evaluated the presence of infection over time in a limited area. Therefore, this study aimed to assess the seroprevalence of N. caninum in a dairy cattle farm in Southeastern Romania where a previously study reported about the cattle exposure to N. caninum infection. A total of 85 cattle, assigned in four groups, according to the reproductive status (repeated breeding syndrome, history of abortion, pregnant cows, and fresh cows) were tested for the presence of N. caninum antibodies, using an indirect ELISA test. The overall seroprevalence was 28.23% (95% CI: 18.67–37.80). According to the reproductive groups, the seroprevalence varied between 19.04% (95% CI: 2.25-35.84) in fresh cows to 38.09% (95% CI: 17.32 - 58.87) in cows with repeated breeding syndrome. By these findings new information is provided about the disease's presence in the studied farm, implying that infection with N. caninum may be the primary cause of reported reproductive disorders, as well as about the infection's persistent nature.

Originals or local replicas? The techno-functional analysis of the disc-shaped antler cheekpieces from the Bronze Age settlement at Sărata Monteoru, south-eastern Romania

Originals or local replicas? The techno-functional analysis of the disc-shaped antler cheekpieces from the Bronze Age settlement at Sărata Monteoru, south-eastern Romania

Boutonneuse Fever in Southeastern Romania.

Boutonneuse fever (BF) is an eruptive disease and is classified as a spotted fever, which is endemic in the Mediterranean basin (i.e., Marseille fever or Mediterranean spotted fever) and the Black Sea, caused by Rickettsia conorii, with dog ticks being a vector (i.e., Rhipicephalus sanguineus). In Romania, although the first reported outbreak of BF occurred during the summer of 1931 in Constanta, the disease was discovered in 1910. Although the disease has occurred most frequently in the two counties of the Dobruja region (Constanta and Tulcea), a region of the Balkan Peninsula, during the last few years, other counties in southeastern Romania have started to report BF cases. In a period of 9 years, 533 cases were registered in Constanta county, while in a period of 11 years, 339 cases were registered in Bucharest county. In this review, we describe the bacterial tick-borne disease caused by R. conorii in southeastern Romania, focusing on its history and epidemiology, pathophysiology, clinical aspects, diagnosis, treatment and preventive measures in the context of climate changes. Although R. conorii is the principal etiologic agent of BF in southeastern Romania, we should take into consideration that other Rickettsia spp. could be present and involved in disease transmission.

CMV and HIV Coinfection in Women from a Region in Eastern Europe.

(1) Background: Human cytomegalovirus (CMV) infection is one of the most frequent opportunistic infections in immunosuppressed patients. Romania has one of the highest incidences of patients living with human immunodeficiency virus (HIV) which determines an immunosuppressive state. The aim of this study was to establish the prevalence of CMV infection among women living with HIV in Southeastern Romania and also to evaluate and correlate antiretroviral therapy (ART) with CD4 level and CMV disease evolution. (2) Methods: Seventy women living with HIV from Southeastern Romania were screened for CMV infection using antigen quantification. Of these, 50 were included in the study. First, the patients filled out a questionnaire regarding social conditions and other associated diseases. Then, we explored the statistical correlations between the data and HIV status, CD4+ cell counts, viral load, and antiretroviral therapy (ART). (3) Results: Median age of the patients was 33 years. Twenty-nine cases were diagnosed with HIV after sexual life beginning and 21 before. Most of the patients had a CD4 level over 200 cells/µL. ART duration in the CD4 under 200 cells/µL group was a bit longer than that in the CD4 over 200 cells/µL group. Forty-one patients had undetectable viremia. CD4 average value in the lot of patients with undetectable viremia was 704.71 cells/µL and in the lot with detectable viremia was 452.44 cells/µL. Viremia values correlated negatively with CD4 level. A positive correlation between IgG CMV values and ART therapy length was identified. A negative significant correlation between values of IgG CMV and values of CD4 was identified. CD4 value correlated negatively with IgG CMV values and with CMV avidity. (4) Conclusions: IgG CMV values had a weak positive correlation with ART therapy length, and a negative statistically significant correlation with values of CD4. CMV avidity has a negative correlation with CD4 value.

Mid-to late Holocene vegetation and environmental change at local and regional scales based on a multi-proxy analysis of the upper Danube Delta, Romania

The Danube Delta is Europe's largest wetland system and of unique biogeographical character. Whilst its geomorphological evolution is relatively well-known its ecological history is poorly understood, including the history of human impact, as a result of the dynamic nature of deltaic systems, and the scarcity of reliably dated successions. In this paper, we report a multi-proxy record of palynology, sediment texture, geochemistry, and ostracods from the upper reaches of the Danube Delta. We use these data to reconstruct a mid-to-late Holocene history of vegetation and environmental changes at local and regional scales, which serve as a model for understanding baseline conditions in delta apex regions. From 7500 to 6200 ​cal ​yr BP, the study site was part of an inter-distributary channel-levee system connected to coastal lagoons and sensitive to sea-level fluctuations but transitioned to a partially-disconnected lacustrine environment after 5700 ​cal ​yr BP. These geomorphologically-driven landscape changes strongly influenced the pollen source area making it complex to interpret palynoassemblages. Prior to 5700 ​cal ​yr BP, palynoassemblages were predominantly river-transported, reflecting widespread hinterlands, and providing information for the regional reconstruction of the vegetation history in the northern Dobrogea region. These data reveal the early presence of Carpinus and Fagus in southeastern Romania, prior to their spread through the Carpathian Basin. After 5700 ​cal ​yr BP, initiated by a slowdown in the relative rate of rise of the Black Sea, peat accumulation commenced within the shallow lake depocentre and airborne pollen became the dominant source. These pollen data record an expansion of herbaceous taxa and highly diverse marsh and aquatic taxa. Tree cover became dominated by Quercus, with low percentages of Carpinus, Betula, Ulmus, and Tilia. The earliest pollen evidence indicative of human impact commences c. 6500 ​cal ​yr BP) and, after 3200 ​cal ​yr BP, the decline in Quercus and a synchronous rise in archaeological artifacts, points to an opening of the landscape by forest clearance and an enlargement of arable areas. In particular, clearance and agriculture during the development of the Hallstattian Babadag culture (c. 3200-2800 ​yr BP) reflects an increase in population and settlement density during that late interval. Our paper comprises the first mid-to-late Holocene palynological record for the Danube Delta and highlights how human activity has profoundly altered the dryland region, creating landscapes comparable to today from around 3200 years BP, while the local deltaic landscape retained its character over the last six millennia.

Comparative Evaluation of the Clinical Severity of COVID-19 of Vaccinated and Unvaccinated Patients in Southeastern Romania in the First 6 Months of 2022, during the Omicron Wave.

(1) Background: The pandemic wave produced by SARS-CoV-2 Omicron was characterized by milder clinical forms and high contagiousness. The vaccination rate against COVID-19 in Romania was approximately 42%. (2) Objectives: Comparison of the clinical severity in vaccinated patients compared to unvaccinated ones. (3) Methods: A retrospective cohort study was conducted on a group of 699 adult patients confirmed with COVID-19 who presented in the "Sf. Cuvioasa Parascheva" Infectious Diseases Clinical Hospital of Galati, Romania, between 1 January 2022 and 30 June 2022. The study compared the need for hospitalization, reinfections, demographic and comorbidity data, clinical and paraclinical parameters from the initial evaluation, and the ratio of unfavorable developments on subgroups chosen according to the vaccination status. (4) Results and Conclusions: Our study reveals that unvaccinated patients required hospitalization in 54.68% of cases, while fully vaccinated patients had a hospitalization rate of 40.72%, which was significantly lower than that of the unvaccinated group (p = 0.01); patients who received a booster dose had a hospitalization rate of 27.84% (p < 0.01, significantly lower than unvaccinated individuals; p = 0.01, significantly lower than fully vaccinated individuals); and among the four patients who received four doses, none required hospitalization. From the analysis of the two subgroups of hospitalized patients, we observed a significantly higher prevalence of radiological lesions, such as pulmonary opacities in the group of unvaccinated patients and a higher average duration of hospitalization, and serum values of D-dimers and blood-sugar at admission were significantly higher in unvaccinated patients. The higher presence of these parameters, which are indicators of severe progression in clinical studies, in the group of unvaccinated patients suggests the need to include them in the initial evaluation of the unvaccinated patients with COVID-19.The cumulative share of deaths and transfers in the ICU was higher in the group of unvaccinated patients, but the difference between the groups had no statistical significance. This study draws attention to the possibility of severe clinical forms among both vaccinated and unvaccinated populations, especially in the elderly and in patients with multiple comorbidities.

Trichinella proficiency testing in Southeastern European countries

In some Southeastern European (SEE) countries, like Serbia, Romania, and Bulgaria, trichinellosis is one of the most important foodborne zoonotic diseases. In those countries, EU regulation and local authorities require the staff of laboratories performing official controls on meat to be properly trained and to check their competence by participating regularly in proficiency testing (PT). PTs are organized by National Reference Laboratories for Trichinella of each country and involve all official laboratories testing meat. In Romania and Bulgaria, the organization of PT for the detection of Trichinella larvae in meat by Magnetic Stirrer Method (MSM) started in 2012. In Croatia and Serbia PT was first organized in 2015 and 2017, respectively. This study presents data on the performance obtained by official laboratories of SEE countries that organize PT at national level and compares the performance obtained by laboratories belonging to different countries. Results suggest that the constant participation in PT leads to an increase in the performance of participating laboratories by positively affecting the staff accuracy in sample testing by MSM. Since the percentage of recovered larvae was in some cases suboptimal (<80%) and occasionally very poor (<40%), there is room for improvement. The regular participation in PT by laboratories involved in official controls on meat intended for human consumption is fundamental to guarantee consumer safety.

Novel in-situ observations on the diel surface activity of the Romanian hamster, Mesocricetus newtoni (Rodentia, Cricetidae), during pup rearing

The Romanian hamster Mesocricetus newtoni is a threatened and cryptic species, with a distribution limited to North-eastern Bulgaria and South-eastern Romania (Dobruja region). Current literature does not provide sufficient data regarding diel activities and behavior of this species, especially under field conditions. In this context, one female M. newtoni, during the rearing period of her four pups, was monitored for ten days using camera traps. For each documented activity, the date, time, age of the individual, type of activity, and duration were recorded. In total, from the 26140 media files, 72968 seconds were recorded as active behaviors. Data analysis revealed that diurnal activity was predominant and documented for all monitored days; however, the animals spent more time per each activity during the night. The nocturnal activity was present but erratic, without significant patterns. No significant differences were observed in the median duration of activities of the adult compared to the immature individuals. Both the adult female and pups spent time exploring the burrow entrance, especially during the day, but immatures were more reserved to leave the burrow than the female. The female used four burrows for rearing the pups. During this time, food storage behaviors and transferring of immatures between burrows, as well as the weaning of the pups were documented. This study demonstrates that M. newtoni exhibits significant diurnal activity, at least in certain ecological and physiological conditions.

Preliminary observations on the food availability and diet composition of the Romanian hamster Mesocricetus newtoni (Rodentia, Cricetidae)

The Romanian hamster Mesocricetus newtoni is endemic to South-Eastern Romania (Dobruja region) and North-Eastern Bulgaria, and prefers steppic, arid, uncultivated habitats, but is also found in alfalfa fields, less often in corn cultures and field-protecting vegetation belts. Previous studies provide only limited data regarding the diet of this cryptic species. The diet of one family group (one female and four pups) was monitored for 10 days, during the vernal season. The data was collected using camera traps and direct observations, supplemented by photos. Plants were identified at the species level. A vegetation survey was performed in a radius of 25 m2 around the burrows using the pratological method, estimating a percentage of each plant species present in the habitat. A number of 16 plants were identified as part of the diet of the Romanian hamster, mostly herbaceous species within the Brassicaceae family. According to the observations, Erodium cicutarium (Geraniaceae) and Papaver rhoeas (Papaveraceae) were consumed most frequently, followed by Descurainia sophia (Brassicaceae), Sisymbrium orientale (Brassicaceae), Medicago minima (Fabaceae) and Buglossoides arvense (Boraginaceae). In the studied area,&amp;nbsp;38 plant taxa were determined and characterized as ruderal and segetal (weeds of arable land) plants, mostly native to Eurasia. The purpose of this study is to provide novel data on the diet composition of the Romanian hamster in accordance with plant taxa availability and to emphasize the importance of conservation measures of its preferred habitats, which are crucial to the survival of the species.

Detection and Molecular Characterization of Canine Babesiosis Causative Agent Babesia canis in Naturally Infected Dogs in the Dobrogea Area (Southeastern Romania).

Canine babesiosis is an emerging tick-borne disease of major veterinary concern in Europe. Its prevalence has increased in the last two decades and is spreading rapidly toward the north. The aim of this study was to investigate the genetic diversity of Babesia spp. strains isolated from naturally infected dogs in a tick-endemic area (Dobrogea) in southeastern Romania. For this purpose, a total of twenty-three samples from dogs diagnosed with various clinical forms of babesiosis, evaluated by means of clinical history, physical examination, and hematological tests, were subjected to a molecular investigation using PCR, sequencing analysis, and genetic characterization. A microscopic examination of thin Diff-quick-stained blood smears revealed large intra-erythrocytic Babesia piroplasms in all dogs. The PCR and sequencing analysis results indicated the presence of Babesia canis in 22 dogs (95.7%) and Babesia vogeli in 1 dog (4.3%). Among the B. canis isolates, two genotypes were distinguished based on two nucleotide substitutions (GA→AG) observed in the 18S rRNA gene sequences (at positions 609 and 610), with the AG genotype predominating (54.5% of samples), while the GA variant was identified in 9.1% of samples. In the remaining isolates (36.4%), both variants were identified. The B. vogeli-positive dog also tested positive for antibodies against Ehrlichia canis and displayed severe disease. This study reports, for the first time, the presence of genetically heterogenic B. canis strains in dogs with clinical babesiosis in Romania. These findings provide a basis for future studies on the relationship between the genetic structure of the causative agents of canine babesiosis in Romania and the course of the disease.

First Detection and Molecular Characterization of Usutu Virus in Culex pipiens Mosquitoes Collected in Romania.

Usutu virus (USUV) is an emergent arbovirus in Europe causing mortality in bird populations. Similar to West Nile virus (WNV), USUV is maintained in sylvatic cycles between mosquito vectors and bird reservoirs. Spillover events may result in human neurological infection cases. Apart from indirect evidence provided by a recent serological study in wild birds, the circulation of USUV in Romania was not assessed. We aimed to detect and molecular characterize USUV circulating in mosquito vectors collected in South-Eastern Romania-a well-known WNV endemic region-during four transmission seasons. Mosquitoes were collected from Bucharest metropolitan area and Danube Delta, pooled, and screened by real-time RT-PCR for USUV. Partial genomic sequences were obtained and used for phylogeny. USUV was detected in Culex pipiens s.l. female mosquitoes collected in Bucharest, in 2019. The virus belonged to Europe 2 lineage, sub-lineage EU2-A. Phylogenetic analysis revealed high similarity with isolates infecting mosquito vectors, birds, and humans in Europe starting with 2009, all sharing common origin in Northern Italy. To our knowledge, this is the first study characterizing a strain of USUV circulating in Romania.

Tentative indicators of malaria in archaeological skeletal samples, a pilot study testing different methods

ObjectiveThis study attempts to integrate multiple methods to investigate the presence of malaria in human skeletal samples from an archaeological context. Materials33 well preserved human remains originating from a 17th-century archaeological site in southeastern Romania. MethodsThe human bone samples were analyzed using rapid diagnostic tests for malaria antigens and PCR amplification of Plasmodium falciparum apical membrane antigen 1. A preliminary test was performed to identify and briefly characterize the presence of hemozoin using a combination of TEM imaging and diffraction. ResultsThe rapid diagnostic tests indicated that more than half of the examined samples were positive for Plasmodium antigens, but no traces of the parasites’ genetic material were detected despite repeated attempts. The TEM images indicated that hemozoin might be a promising diagnostic marker of malaria in ancient bones. ConclusionsThe indisputable identification of malaria in the analyzed archaeological population was not possible as none of the applied methodological strategies turned out to be straightforward. SignificanceThis study reinforces the intricacy and limitations of unequivocally identifying malaria in past populations and sets the stage for future studies on such life-threatening infectious disease in a geographical space, which is currently underrepresented in the bioarchaeological literature. LimitationsThe low sample size and the lack of consistency across all assays hindered understanding the role of malaria in the studied population. Suggestions for further researchFurther thorough multidisciplinary approaches on malaria detection in ancient settlements would be appropriate to inform our knowledge of its origins, frequency, and pathogen changes over centuries.

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.