Seroprevalence of Anti-Hepatitis E Virus Antibodies among Patients from a Tertiary Hospital from Northeast Romania.

Background and Objectives. Being an enterically transmitted pathogen with a growing prevalence in developed countries, hepatitis E virus (HEV) infection remains an underdiagnosed disease in Eastern Europe. As far as Romania is concerned, only a few studies address this issue. Our goal was to estimate the prevalence of serum anti-HEV IgA/IgM/IgG antibodies in a group of patients admitted to the Clinical Hospital for Infectious Diseases "St. Parascheva" Iasi. Materials and Methods. The cross-sectional study consisted of enrollment of 98 patients admitted to the clinic for COVID-19 over a period of three months in 2020. Results. The median age in our study was 73 years, with an equal gender ratio and with a predominance of people from the urban environment (75%). The overall HEV antibody seroprevalence was 12.2%. The main risk factors associated with HEV infection were consumption of water from unsafe sources (58.3% HEV-positive patients vs. 26.7% HEV-negative patients, p = 0.026) and improperly cooked meat (58.3% HEV-positive patients vs. 23.2% HEV-negative patients, p = 0.01). Zoonotic transmission was an important criterion in our study, with patients reporting contact with pigs, poultry, rats, or other farms animals, but no significant differences were found between HEV antibody positive and negative groups. Conclusions. The seroprevalence rate of HEV antibodies was similar to other previous reports from our area but higher than in most European countries. The fact that HEV antibodies were detected in patients without identifiable risk factors for hepatitis E is evidence of subclinical infection as a silent threat.

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.

Clinical significance of early IgA anti-SARS-CoV-2 antibody detection in patients from a Romanian referral COVID-19 hospital.

Controlling the spread of coronavirus disease 2019 (COVID-19) includes institute isolation, quarantine measures and appropriate clinical management, which all require effective screening, diagnostic and prognostic tools. The present study aimed to analyze severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-specific immunoglobulin (Ig)A detection and determine the potential association with the clinical course of COVID-19 and the levels of inflammation. In the present study, the presence of IgA and IgG SARS-CoV-2 antibodies in 75 consecutive patients with confirmed COVID-19 infection was investigated. No significant differences were found between the IgA positive and negative groups, regarding the presence of symptoms, haematological and inflammatory variables, or the presence of pneumonia. In the majority of cases, antibody detection was comparable, for example, 79.7% of patients in the IgA positive group exhibited both types of antibodies, while 80.9% of patients in the IgA negative group were also IgG negative. A total of four patients in the IgA negative group presented with anti-SARS-CoV-2 IgG antibodies. Early detection of IgA was more frequent in patients who later developed severe forms of the disease. In addition, the IgG SARS-CoV-2 antibody response was higher in patients with the severe form of the disease.

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.

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.

Molecular Evidence of Rickettsia spp., Anaplasma phagocytophilum, and "Candidatus Neoehrlichia mikurensis" in Ticks from Natural and Urban Habitats in Eastern Romania.

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 Iasi 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 Iasi 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.

Evidence of hepatitis E infection in swine and humans in the East Region of Romania.

OBJECTIVE: Swine hepatitis E virus (HEV) is considered to be a new zoonotic agent due to its close genomic resemblance to the human HEV. The aim of this study was to determine human HEV seroprevalence in eastern Romania and to characterize circulating swine HEV sequences. METHODS: Serological investigations of human serum samples were done using a commercial ELISA kit (MP Biomedicals). Swine faecal samples were tested to detect the HEV ORF2 sequence by nested reverse transcription PCR. RESULTS: One hundred and forty-eight human serum samples were tested for anti-HEV IgG of which 22 were found to be positive. Fresh swine faeces (pools) were collected from five farms in eastern Romania. Six out of 19 pooled samples were positive for HEV RNA. Phylogenetic analysis based on alignment of the ORF2 sequence indicated that the Romanian swine HEV isolates belonged to genotype 3. CONCLUSIONS: This is the first study showing HEV to be present in Romanian pig herds and that the human population is exposed.

West Nile virus reemergence in Romania: a serologic survey in host species.

The presence of West Nile virus (WNV) in humans has been known in Romania since the 1950s; the 1996 epidemics emphasized the reemergence potential of WNV in Romania. Serological surveys made on susceptible species, known as good sentinels or reservoir hosts, e.g., horses, wild and domestic birds were undertaken from 2006-2011. Our results corroborated incidence data in human patients and other recent seroprevalence studies in animals, and should partially clarify the emergence of WNV in the eastern rural territories of Romania. It also highlighted risk zones for endemic WNV infection in Romania.