An Assessment of West Nile and Usutu Viruses' Seroprevalence in Hospitalized Patients: A Preliminary Study on Flavivirus Exposure in Eastern Romania.

WNV and USUV are closely related epornitic flaviviruses transmitted by Culex mosquitoes which can cause febrile and neurodegenerative disease in humans. The impact of both viruses on public health has increased in the recent decades. AIM: The aim of the study was to evaluate the seroprevalence of WNV and USUV in hospitalized patients from eastern Romania who did not show symptoms corresponding to the case definition. METHODS: Human blood samples from the hospitalized patients were collected in 2015 and from April to September 2019 in Iasi County, Romania. The samples were screened by ELISA for anti-WNV IgG, IgM, and anti-USUV IgG antibodies. RESULTS: A cumulative seroprevalence of 3.4% was recorded for anti-WNV IgG antibodies and 9.1% for anti-WNV IgM. No sample was positive for anti-USUV antibodies. CONCLUSION: The cumulative seroprevalence observed provides support for the consideration of WNV as being endemic in the east of Romania. The absence of anti-USUV antibodies may be related to cross-reactivity and cohort size, thus, USUV should be considered in clinical practice and become an objective for active surveillance in Romania.

Antimicrobial Activity of Artemisia dracunculus Oil-Loaded Agarose/Poly(Vinyl Alcohol) Hydrogel for Bio-Applications.

In this study, the potential use of Artemisia dracunculus essential oil in bio-applications was investigated. Firstly, the phytochemicals from Artemisia dracunculus were analyzed by different methods. Secondly, the Artemisia dracunculus essential oil was incorporated into the hydrogel matrix based on poly(vinyl alcohol) (PVA) and agar (A). The structural, morphological, and physical properties of the hydrogel matrix loaded with different amounts of Artemisia dracunculus essential oil were thoroughly investigated. FTIR analysis revealed the successful loading of the essential oil Artemisia dracunculus into the PVA/A hydrogel matrix. The influence of the mechanical properties and antimicrobial activity of the PVA/A hydrogel matrix loaded with different amounts of Artemisia dracunculus was also assessed. The antimicrobial activity of Artemisia dracunculus (EO Artemisia dracunculus) essential oil was tested using the disk diffusion method and the time-kill assay method after entrapment in the PVA/A hydrogel matrices. The results showed that PVA/agar-based hydrogels loaded with EO Artemisia dracunculus exhibited significant antimicrobial activity (log reduction ratio in the range of 85.5111-100%) against nine pathogenic isolates, both Gram-positive (S. aureus, MRSA, E. faecalis, L. monocytogenes) and Gram-negative (E. coli, K. pneumoniae, S. enteritidis, S. typhimurium, and A. salmonicida). The resulted biocompatible polymers proved to have enhanced properties when functionalized with the essential oil of Artemisia dracunculus, offering opportunities and possibilities for novel applications.

Clonal Dissemination of Extended-Spectrum Cephalosporin-Resistant Enterobacterales between Dogs and Humans in Households and Animal Shelters of Romania.

Faecal carriage of extended-spectrum cephalosporin-resistant (ESC-R) Enterobacterales in healthy pets is a concerning issue. This study aimed to determine the prevalence, genetic background, and potential for interspecies transmission of these bacteria between dogs and humans within the same household (HH) or shelter environment in Romania. Faecal samples (n = 263) collected from healthy dogs (n = 102), their owners (n = 32), as well as dogs (n = 110) and staff (n = 19) from dog shelters, were screened for ESC-R carriage. Clonal relatedness of canine and human Escherichia coli isolates was established using Fourier Transform Infrared Spectroscopy (FTIR), followed by Illumina WGS of selected isolates. The highest prevalence of ESC-R Enterobacterales faecal carriage was identified in staff working at dog shelters (78.9%), followed by dogs from households (44.11%), dog owners (43.7%), and dogs from shelters (27%). FTIR identified 15 clusters of closely related E. coli isolates, including dog and human isolates from the same environment. Co-carriage of ESC-R isolates in both the dog and owner was identified in 12 HHs (37.5%), with two HHs (6%) having both the owner and dog carrying isolates with identical FTIR spectra, phylogroup, resistance genes, and Inc plasmids. Major ExPEC lineages such as ST127, ST10, ST155, and ST88 were detected in human and dog isolates. Our study revealed a high prevalence of faecal ESC-R E. coli carriage in both dogs and humans from Romanian households and shelters, where bidirectional clonal transmission between humans and dogs is likely. Furthermore, we identified ESC-R Enterobacterales co-carriage in people and dogs sharing the same environment using FTIR, demonstrating its value in AMR surveillance for humans and animals.

Seroprevalence of Crimean-Congo Hemorrhagic Fever Among Small Ruminants from Southern Romania.

Crimean-Congo hemorrhagic fever (CCHF) is a tick-borne disease that can be contracted by direct contact with viremic animals or humans. Domestic animals are accidental hosts and contribute to the spread and amplification of the virus. The main objective of this study was to provide updated information related to CCHF virus (CCHFV) infection in Southern Romania by assessing the seroprevalence of CCHF in small ruminants (sheep and goats) using a double-antigen sandwich enzyme-linked immunosorbent assay and by detection of CCHFV in engorged ticks and serum samples using real-time RT-PCR. The overall seroprevalence of CCHF in small ruminants was 37.7% (95% CI 31.7 to 43.7). No statistical seroprevalence difference was observed between the two species of ruminants (p = 0.76), but a significant difference was established between the locations (p < 0.01). No CCHFV RNA was detected in tick pools and small ruminant's sera tested by real-time RT-PCR, although the high seroprevalence to CCHFV among ruminants indicates that CCHV or a closely related virus circulates in Southern Romania.

Thyroid Cancer Diagnostics Related to Occupational and Environmental Risk Factors: An Integrated Risk Assessment Approach.

There are still many questions remaining about the etiopathogenesis of thyroid cancer, the most common type of endocrine neoplasia. Numerous occupational and environmental exposures have been shown to represent important risk factors that increase its incidence. Updated information about thyroid cancer diagnostics related to occupational and environmental risk factors is reviewed here, considering an integrated risk assessment approach; new data concerning thyroid cancer etiology and pathogenesis mechanisms, diagnostic biomarkers and methodologies, and risk factors involved in its pathogenesis are presented. A special emphasis is dedicated to specific occupational risk factors and to the association between environmental risk agents and thyroid cancer development. The occupational environment is taken into consideration, i.e., the current workplace and previous jobs, as well as data regarding risk factors, e.g., age, gender, family history, lifestyle, use of chemicals, or radiation exposure outside the workplace. Finally, an integrative approach is presented, underlying the need for an accurate Risk Assessment Matrix based on a systematic questionnaire. We propose a complex experimental design that contains different inclusion and exclusion criteria for patient groups, detailed working protocols for achieving coherent and sustainable, well-defined research stages from sample collection to the identification of biomarkers, with correlations between specific oncometabolites integrated into the Risk Assessment Matrix.

Common European Origin of Hepatitis E Virus in Human Population From Eastern Romania.

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.

Genetic Features of mcr-1 Mediated Colistin Resistance in CMY-2-Producing Escherichia coli From Romanian Poultry.

Colistin is a last resort antibiotic used for the treatment of human infections associated with carbapenemase-producing Enterobacteriales. Here, we evaluated the occurrence of mcr-1 and -2 plasmid-mediated colistin resistance in colistin and/or carbapenem resistant human clinical Enterobacteriales and other gram-negative bacteria (n = 543) as well as third generation cephalosporin-resistant (3GCR) Escherichia coli isolates from poultry abattoir workers (n = 15) and poultry fecal samples (n = 92) collected from two geographically separate abattoirs in Romania. which revealed that mcr-1 was present within four sequence types (STs): ST744 (n = 7), ST57 (n = 7), ST156 (n = 2), and ST10 (n = 1). Within STs, serotypes were conserved and, notably, all except one of the mcr-1-positive isolates were found to exhibit fluoroquinolone-resistance (FQR) associated SNPs in both gyrA and parC. While there were variations in genotypes, all isolates belonging to ST744, ST57, and ST156 were rich in resistance determinants, carrying aminoglycoside-modifying enzymes genes, sulfonamide resistance gene bla TEM- 1 as well as bla CMY- 2 AmpC ß-lactamase resistance genes. They also exhibited high similarity in carriage of virulence genes; ST10, however, only carried the mcr-1 gene. Whole genome sequencing (WGS) analysis also revealed that although the mcr-1 gene was identified in a diverse population of E. coli, two STs (ST57 and ST744) predominated and interestingly, were found in isolates across both abattoirs providing evidence for clonal transmission. Also, two main genomic contexts of mcr-1 isolates were revealed with all ST57 isolates harboring the mcr-1 gene between two copies of ISApl1 (or the Tn6330 transposon) whilst a common mcr-1 containing scaffold, highly similar to IncX type mcr-1-bearing plasmids (pWI2-mcr, Accession number: LT838201), was present among mcr-1 isolates of varying phylogenetic backgrounds (ST10, ST744 and ST156). The high prevalence of the mcr-1 gene in poultry E. coli isolates with co-resistance to cephalosporins and quinolones, in a country where antimicrobial use in food production species is poorly regulated, is concerning and the findings from this study should lead to better surveillance of antimicrobial resistance (AMR) in food-production animals in Romania.