Toxoplasma gondii genotypes and frequency in domestic cats from Romania.

BACKGROUND: Toxoplasma gondii is a zoonotic protozoan parasite with a heteroxenus life cycle that involves felids as the definitive hosts and any warm-blooded animal, including humans, as intermediate hosts. Cats are key players in parasite transmission as they are capable of shedding high numbers of oocysts in their feces that contaminate the environment. METHODS: The study was performed on 31 domestic cats (31.23 ± 27.18 months old) originating from rural and urban areas (5.17:1) in the center and north-west Romania. Feces (n = 31), blood (n = 28), and heart samples (n = 27) were collected. Fecal samples were analyzed by flotation technique, and PCR (529 bp repetitive element). Fecal samples with T. gondii oocysts were bioassayed in mice. Serum samples were analyzed by modified agglutination test and ImmunoComb for the detection of specific anti-T. gondii IgG antibodies. Heart samples were bioassayed in mice, and analyzed by PCR. Toxoplasma gondii positive samples were genotyped by nPCR-RFLP targeting eleven genetic loci (SAG1, SAG2, alt-SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico). RESULTS: Toxoplasma gondii oocysts were found in 2 out of 31 fecal samples collected from a 3-months old stray kitten, and a 4-years old female. In total, 17 out of 27 sera were positive for T. gondii IgG antibodies. The antibody titers in MAT ranged from 1:6 to 1:384. Toxoplasma gondii DNA was detected in 7 out of 27 heart samples, and four of them were positive also by bioassay. Six T. gondii DNA samples from bioassayed mice could be assigned to ToxoDB PCR-RFLP genotype #1 or #3 (Type II) and one T. gondii DNA from heart digest to genotype #2 (Type III). Both of these genotypes are common in Europe. CONCLUSIONS: Our results revealed that the infection with T. gondii is still high in cats from Romania. The oocysts shedded by these cats represent an important source of infection for intermediate hosts, including humans. Further studies on a wider range of cases are necessary for a more exhaustive definition of the T. gondii genotypes circulating in Romania.

VEGF Polymorphisms (VEGF-936 C/T, VEGF-634 G/C and VEGF-2578 C/A) and Cardiovascular Implications in Long COVID Patients.

The COVID-19 pandemic has raised awareness of the virus's long-term non-pulmonary consequences. This study examined the relationship between genetic polymorphisms of VEGF and cardiac dysfunction and subclinical atherosclerosis in patients recovering from COVID-19. This study included 67 patients previously diagnosed with COVID-19. VEGF-936C/T, VEGF-634G/C, and VEGF-2578C/A statuses were determined. Conventional echocardiography and arterial parameters assessments were performed at inclusion and at six months after the first assessment. For VEGF-936C/T, dominant and over-dominant models showed a significant increase in ejection fraction at six months after COVID (p = 0.044 and 0.048) and was also a predictive independent factor for the augmentation index (ß = 3.07; p = 0.024). The dominant model showed a rise in RV-RA gradient (3.702 mmHg) (p = 0.028 95% CI: 0.040-7.363), with the over-dominant model indicating a greater difference (4.254 mmHg) (p = 0.025 95% CI: 0.624-7.884). The findings for VEGF-634G/C were not statistically significant, except for a difference in TAPSE during initial evaluation, using the codominant model. For VEGF-2578C/A, a difference in ventricular filling pressure (E/E'ratio) was best described under the recessive model. Our research suggests that the VEG-936C/T genotype may impact the baseline level and subsequent changes in cardiac function and subclinical atherosclerosis. These findings offer valuable insights into the complex correlation between genetic polymorphisms and cardiovascular disfunction in long COVID patients.

The Impact of Long COVID on the Quality of Life.

Background and Objectives: The term long COVID refers to patients with a history of confirmed COVID-19 infection, who present symptoms that last for at least 2 months and cannot be explained by another diagnosis. Objectives: The present study aims to determine the most common symptoms of the long COVID syndrome and their impact on the quality of life. Materials and Methods: A prospective observational study was conducted on patients diagnosed with mild and moderate COVID-19 (based on a positive SARS-CoV-2 molecular diagnostic or rapid antigen test and severity form definition) at the Clinical Hospital of Infectious Diseases, Cluj-Napoca, Romania. Clinical examinations with detailed questions about symptoms were performed at the time of the diagnosis of COVID-19 and the six-month follow-up. Two years after COVID-19 infection, patients were invited to complete an online quality-of-life questionnaire regarding long COVID symptoms. Results: A total of 103 patients (35.92% males) with a mean age of 41.56 ± 11.77 were included in this study. Of the total number of patients, 65.04% presented mild forms of COVID-19. Data regarding the vaccination status showed that 83.5% were vaccinated against SARS-CoV-2. The most common symptoms at diagnosis were cough (80.6%), fatigue (80.4%), odynophagia (76.7%), and headaches (67.6%), with female patients being statistically more likely to experience it (p = 0.014). Patients with moderate forms of the disease had higher levels of both systolic (p = 0.008) and diastolic (p = 0.037) blood pressure at diagnosis, but no statistical difference was observed in the 6-month follow-up. The most common symptoms at 2 years (in 29 respondent subjects) were represented by asthenia (51.7%), headache (34.5%), memory disorders (27.6%), abdominal meteorism (27.6%), and arthralgia (27.6%). In terms of cardiovascular symptoms, fluctuating blood pressure values (20.7%), palpitations (17.2%), and increased heart rate values (17.2%) were recorded. Conclusions: If at the time of diagnosis, the most frequent manifestations of the disease were respiratory, together with headache and fatigue, at re-evaluation, asthenia, decreased effort tolerance, and neuropsychiatric symptoms prevailed. Regarding the cardiovascular changes as part of the long COVID clinical picture, some patients developed prehypertension, palpitations, and tachycardia.

Immunohistochemistry and real-time PCR as diagnostic tools for detection of Borrelia burgdorferi sensu lato in ticks collected from humans.

The objective of this study was to evaluate different methods used for detection of Borrelia burgdorferi sensu lato (s.l.) in ticks: immunohistochemistry followed by focus floating microscopy (FFM) and real-time polymerase chain reaction (real-time PCR) targeting the ospA and hbb genes. Additionally, an optimized ospA real-time PCR assay was developed with an integrated internal amplification control (IAC) for the detection of inhibition in the PCR assay and was validated as an improved screening tool for B. burgdorferi. One hundred and thirty-six ticks collected from humans in a hospital from Cluj-Napoca, Romania, were investigated regarding genus, stage of development and sex, and then tested by all three assays. A poor quality of agreement was found between FFM and each of the two real-time PCR assays, as assessed by concordance analysis (Cohen's kappa), whereas the agreement between the two real-time PCR assays was moderate. The present study argues for a low sensitivity of FFM and underlines that discordant results of different assays used for detection of B. burgdorferi in ticks are frequent.

Incidence of symptomatic Borrelia burgdorferi sensu lato infection in Romania, 2018-2023.

BACKGROUND: Lyme borreliosis (LB), caused by Borrelia burgdorferi sensu lato (Bbsl), is the most common tick-borne disease in Europe. Although public health surveillance for LB has been conducted in Romania since 2007, the extent of under-detection of Bbsl infections by LB surveillance has not been estimated. We therefore estimated the under-detection of symptomatic Bbsl infections by LB surveillance to better understand the LB burden in Romania. METHODS: The number of incident symptomatic Bbsl infections were estimated from a seroprevalence study conducted in six counties (population 2.3 M) and estimates of the symptomatic proportion and duration of persistence of anti-Bbsl immunoglobulin G (IgG) antibodies. The number of incident symptomatic Bbsl infections were compared with the number of surveillance-reported LB cases to derive an under-detection multiplier, and then the under-detection multiplier was applied to LB surveillance data to estimate the incidence of symptomatic Bbsl infection from 2018 to 2023. RESULTS: We estimate that there were 1968 individuals with incident symptomatic Bbsl infection in the six counties where the seroprevalence study was conducted in 2020, compared with the 187 surveillance-reported LB cases, resulting in an under-detection multiplier of 10.5 (i.e., for every surveillance-reported LB case, there were 10.5 symptomatic incident Bbsl infections). The incidence of symptomatic Bbsl infection in the six counties was 86.9/100,000 population in 2023, similar to the incidence in 2018-2020 (86.0) and higher than in 2021-2022 (40.3). CONCLUSIONS: There is a higher incidence of symptomatic Bbsl infection than is reported through public health surveillance for LB in Romania. Additional efforts are needed to strengthen disease prevention and address the important public health problem of LB.

Prevalence and Genotyping of Water- and Food-Borne Parasitic Protozoans (Giardia duodenalis and Cryptosporidium spp.) in Hospitalized Patients from Northwestern Romania.

Giardia duodenalis and Cryptosporidium spp. are important zoonotic protozoan pathogens that infect the gastro-intestinal tract of numerous vertebrates, including humans, and both parasites are responsible for water- or food-borne outbreaks of disease worldwide. Although, globally, both parasites are highly prevalent, particularly in developing countries, epidemiological data from Romania are scarce, and genotyping has rarely been performed. The aims of the present study were to investigate the occurrence and genetic diversity of G. duodenalis and Cryptosporidium spp. in patients hospitalized in Northwestern Romania in relation to clinical and paraclinical presentation and to identify the relative frequency of non-specific symptoms and potential risk factors. Between June 2022 and January 2024, 426 fecal samples were screened for gastro-intestinal parasites by rapid tests and microscopical examination, further confirmed by PCR and sequencing. Giardia duodenalis was detected and characterized in 12 samples (2.82%), while Cryptosporidium parvum was confirmed in four samples (0.94%). A majority of positive patients were symptomatic and reported nausea and vomiting with a significantly higher frequency compared to negative ones. This study provides new insights into the epidemiological status and clinical implications of gastro-intestinal parasite species and genospecies in Romania that are necessary for an in-depth understanding of the potential zoonotic transmission and improvement of patient care.

Changes in Epidemiology and Antibiotic Prescription of Influenza: Before and after the Emergence of COVID-19.

BACKGROUND: The appearance of COVID-19 had a major impact on healthcare and the epidemiology of other diseases. Following the cessation of non-pharmacologic interventions destined to limit the spread of COVID-19, influenza reemerged. The aim of this study was to compare the pre-pandemic influenza seasons with the influenza seasons after the emergence of the COVID-19 pandemic, and to identify differences in terms of clinical characteristics, risk factors, complications, outcomes, and antiviral and antibiotic treatments. METHODS: We conducted a retrospective cohort study from the Teaching Hospital of Infectious Diseases database in Cluj-Napoca, Romania. We analyzed four pre-pandemic seasons and the seasons after the onset of COVID-19. We included adult patients hospitalized with confirmed influenza between October 2016 and August 2023. Variables such as age, sex, duration of hospitalization, severity, clinical manifestations, comorbidities, and Charlson comorbidity index were assessed. RESULTS: A total of 941 patients were included in the analysis. The percentage of severe influenza was similar in both groups, but mortality from influenza was significantly lower after 2022. Virtually all patients were prescribed antivirals; antibiotic prescriptions decreased in the post-COVID-19 influenza seasons. CONCLUSION: The present study suggests that influenza seasons after 2022 had lower mortality and attenuated clinical presentation.

Serological evidence of human exposure to foodborne zoonotic parasites in Romanian patients and possible association with food habits and hygiene.

Humans may become accidental dead-end hosts for a series of zoonotic foodborne parasites, of which Toxoplasma gondii, Echinococcus spp., Toxocara spp., and Trichinella spiralis are of major public health interest, due to their potential pathological implications. The aims of the study were to evaluate the exposure to these pathogens in north-western Romania, and to investigate their potential association to risk factors. From June 2022 to January 2024, 554 patients admitted to a tertiary hospital in north-western Romania were screened for the presence of IgG antibodies against T. gondii, Echinococcus spp., Toxocara canis, and T. spiralis by ELISA, and potential risks were assessed using a questionnaire. Overall, 225 samples (40.6%) were positive for at least one pathogen. The highest seroprevalence for IgG was found for T. gondii (33.9%), followed by Echinococcus spp. (9.1%), T. spiralis (2.9%), and T. canis (1.1%). For T. spiralis, raw meat consumption was associated with positivity. For T. gondii, increased age, rural environment, contact with cats, consumption of unwashed fruits/vegetables and drinking water from unverified sources were significantly associated to seropositivity. The present study provides new insights into the epidemiological status of zoonotic foodborne parasite in Romania, underlining the need to increase awareness on the importance of water, sanitation and food habits in relation with this neglected pathology.

Burnout, Depression, and Job Stress Factors in Healthcare Workers of a Romanian COVID-19 Dedicated Hospital, after Two Pandemic Years.

The COVID-19 pandemic put unprecedented pressure on all areas of activity, especially healthcare workers. Understanding the psychological response to the pandemic in healthcare workers is an important challenge. This study aims to investigate burnout, depression, and job stress factors in the medical personnel of a COVID-19-dedicated hospital, two years after the beginning of the pandemic. The survey was performed between the fifth and sixth pandemic waves in Romania. Employees of the Clinical Hospital for Infectious Diseases, Cluj-Napoca, completed an online survey using four tools: Maslach Burnout Inventory (MBI), Copenhagen Burnout Inventory (CBI), the Karasek Job factors questionnaire, and the Patient Health Questionnaire-9 (PHQ-9). A total of 114 employees completed the questionnaire (10.83% of total employees). The results showed 100% prevalence of Maslach burnout (56.1% moderate and severe burnout) and 63.1% prevalence of depression. The infectious disease resident doctors had the highest prevalence of burnout scores, depression, and perceived Karasek job demands. The 22- to 30-year-old age group and the group with fewer than ten years of professional experience had a significantly higher prevalence of burnout and depression than older employees or employees with more professional experience. The COVID-19 pandemic continues to have a high impact on the mental health of healthcare workers.

Retrospective Survey of Human Trichinellosis in a Romanian Infectious Diseases Hospital over a Thirty-Year Interval-The Never-Ending Story.

Trichinellosis remains a food-safety risk in Romania due to cultural traditions and food behavior. The aim of the present study was to evaluate the epidemiological, clinical and therapeutical data of all human trichinellosis cases in patients admitted to an Infectious Diseases Hospital from northwestern Romania during a thirty-year interval. Between 1 January 1988 and 31 December 2018, a total of 558 patients were hospitalized with the diagnosis of trichinellosis. The number of cases/year varied between 1 and 86. The source of infection was known for 524 patients, represented by domestic pig meat (n = 484; 92.37%) and wild boar (n = 40; 7.63%). Most patients (410; 73.48%) presented were part of family or group outbreaks. Demographical and clinical data of patients will be presented. Antiparasitic therapy was prescribed in 99.46% of cases, and corticosteroids were prescribed in 77.06% of patients. In total, 48 patients (8.6%) presented complications of trichinellosis: 44 for a single complication (neurological, cardiovascular or respiratory); the others multiple complications. Pregnancy was documented in five patients. No fatalities occurred during the study period. Although the number of hospitalized patients has decreased in the last years, trichinellosis still remains an important public health problem in northwestern Romania.

25 Hydroxyvitamin D Serum Concentration and COVID-19 Severity and Outcome-A Retrospective Survey in a Romanian Hospital.

Interest in the immunomodulatory function of vitamin D has grown since the COVID-19 pandemic started. Our study investigated the possible association between vitamin D deficiency and COVID-19 severity, intensive care needs, and mortality in patients hospitalized with COVID-19. A prospective cohort study was performed on 2342 COVID-19 hospitalized patients between April 2020 and May 2022 in a Romanian tertiary hospital for infectious diseases. A multivariate generalized linear model for binary data was fit with dependent variables: severe/critical form of COVID-19, intensive care need, and fatal outcome as a function of vitamin D deficiency, controlling for age, comorbidities, and vaccination status. More than half of the patients (50.9%) were classified with vitamin D deficiency based on a serum concentration of less than 20 ng/mL. There was a negative association between vitamin D and age. Vitamin D-deficient patients presented with more cardiovascular, neurological, and pulmonary diseases, as well as diabetes, and cancer. In multivariate logistic regression models, vitamin D-deficient patients had higher odds of severe/critical forms of COVID-19 [OR = 1.23 (95% CI 1.03-1.47), p = 0.023] and higher odds of death [OR = 1.49 (95% CI 1.06-2.08), p = 0.02]. Vitamin D deficiency was associated with disease severity and death outcome in hospitalized COVID-19 patients.

Comparison of COVID-19 Severity in Vaccinated and Unvaccinated Patients during the Delta and Omicron Wave of the Pandemic in a Romanian Tertiary Infectious Diseases Hospital.

Romania has a poor uptake of COVID-19 vaccination in its population. The study objectives were to evaluate the differences between vaccinated and unvaccinated hospitalized COVID-19 patients with regard to disease severity, intensive care need, and mortality during the fourth and the fifth wave of the pandemic associated with the Delta and Omicron variants of concern. A retrospective study on a cohort of hospitalized COVID-19 patients was performed in a Romanian tertiary hospital for infectious diseases. Multivariate logistic regression models were built predicting severe/critical COVID-19, intensive care need, and death as a function of vaccination status and adjusted for age, comorbidities, and wave of the pandemic. 2235 COVID-19 patients were included, and vaccination status, as a primary vaccination or a booster dose, was described in 750 (33.5%). Unvaccinated patients were older, with more cardiovascular and endocrine diseases, a longer duration of hospitalization, a higher percentage of severe/critical COVID-19, need for intensive care, and death (p < 0.05). The multivariate logistic regression models adjusted for age and comorbidities showed higher odds ratio for severe/critical COVID-19, intensive care need, and mortality in unvaccinated versus vaccinated patients. Our results support vaccination to prevent severe outcomes associated with COVID-19 due to both variants of concern.

Toxoplasmosis Screening during Pregnancy in a Romanian Infectious Diseases Tertiary Center: Results of a 15 Years Follow-Up Program.

Maternal infection with Toxoplasma gondii during pregnancy may have serious consequences for the fetus. In Romania, screening for toxoplasmosis is included in the first antenatal visit. A retrospective study was performed on all toxoplasmosis antenatal screening patients between May 2008 and February 2023. Twenty-seven thousand one hundred sixty-nine (27,169) pregnant women presented for prenatal screening once (22,858) or several times: during the same pregnancy (209) or during multiple pregnancies (4102). Thirty-one thousand six hundred fifty-eight (31,658) tests for IgM and IgG antibodies were performed. Nine thousand eighty-three (9083) tests (28.69%), corresponding to 7911 women (29.12%), were positive for IgG antibodies. The seroprevalence increased with patients' age, decreased in time intervals, and was more frequently associated with rural residence. At risk for acquiring the infection during the pregnancy were women with negative anti-Toxoplasma IgG antibodies (70.88%), but only 0.9% of them presented for rescreening during the same pregnancy. Acute Toxoplasma infection (ATI) was suspected in 44 patients (0.16%) due to IgG seroconversion and/or low or borderline IgG avidity. A questionnaire follow-up interview was performed, and no congenital toxoplasmosis was identified in children born from mothers with probable ATI. Our study demonstrates poor compliance with the screening program in the Romanian population.

The Role of Wild Boars in the Circulation of Tick-Borne Pathogens: The First Evidence of Rickettsia monacensis Presence.

Most wild mammals can serve as hosts both for tick-borne pathogens (TBPs) and for the ticks themselves. Among these, wild boars, due to their large body size, habitat and life span, show high exposure to ticks and TBPs. These species are now one of the widest-ranging mammals in the world, as well as the most widespread suid. Despite the fact that certain local populations have been decimated by African swine fever (ASF), wild boars are still considered overabundant in most parts of the world, including Europe. Altogether, their long-life expectancy, large home ranges including migration, feeding and social behaviors, wide distribution, overabundance and increased chances of interactions with livestock or humans make them suitable sentinel species for general health threats, such as antimicrobial-resistant microorganisms, pollution and ASF geographical distribution, as well as for the distribution and abundance of hard ticks and also for certain TBPs, such as Anaplasma phagocytophilum. The aim of this study was to evaluate the presence of rickettsial agents in wild boars from two counties in Romania. Among 203 blood samples of wild boars (Sus scrofa ssp. attila) collected during 3 (2019-2022) hunting seasons (September-February), 15 were found positive for tick-borne pathogen DNA. Six wild boars were positive for A. phagocytophilum DNA presence and nine for Rickettsia spp. The identified rickettsial species were R. monacensis (six) and R. helvetica (three). No animal was positive either for Borrelia spp., Ehrlichia spp. or Babesia spp. To the best of our knowledge, this is the first report of R. monacensis in European wild boars, thus adding the third species from the SFG Rickettsia, in the epidemiology of which this wild species may have a role as a reservoir host.

Evolving Clinical Manifestations and Outcomes in COVID-19 Patients: A Comparative Analysis of SARS-CoV-2 Variant Waves in a Romanian Hospital Setting.

The aim of this study was to evaluate differences in the clinical manifestations and outcomes in hospitalized patients with COVID-19 in a single Romanian center during four pandemic waves determined by different SARS-CoV-2 variants of concern (VOCs). A retrospective study on 9049 consecutive hospitalized adult patients was performed between 27 February 2020 and 31 March 2023. The study interval was divided into waves based on national data on SARS-CoV-2 VOCs' circulation. Multivariate logistic regression models were built, predicting death and complications as functions of comorbidities, therapy, wave, severity form, and vaccination status, and adjusted for ages ≥65 years. Pulmonary (pneumothorax/pneumomediastinum, pulmonary embolism) and extrapulmonary complications (liver injury, acute kidney injury, ischemic/hemorrhagic stroke, myocardial infarction, and gastrointestinal bleeding) were present, more frequently in ICU hospitalized patients and with differences between waves. The highest in-hospital mortality was found in patients presenting pneumothorax/pneumomediastinum. All of the evaluated risk factors were significantly associated with death, except for obesity and the Omicron wave. Our study highlights the changing nature of COVID-19 and acknowledges the impacts of viral mutations on disease outcomes. For all four waves, COVID-19 was a severe disease with a high risk of poor outcomes.

Effects of COVID-19 on the Liver and Mortality in Patients with SARS-CoV-2 Pneumonia Caused by Delta and Non-Delta Variants: An Analysis in a Single Centre.

The aim of this study was to ascertain patient characteristics, outcomes, and liver injuries in patients infected with different SARS-CoV-2 variants. Data from consecutive adult patients with severe/critical COVID-19 admitted to our hospital during the peak month of the Delta wave were compared to the ancestral, Alpha, and Omicron waves. The dataset of 551 hospitalized patients was similar in the Delta/non-Delta waves. At admission and discharge, the median aminotransferase levels were normal or slightly increased. During the Delta wave (172 vs. 379 non-Delta patients), more patients died (OR 1.69, 95%CI 1.09-2.56) or had liver injury at discharge (alanine aminotransferase, ALT ≥ 2 ULN) (OR 1.97, 95%CI 1.08-3.54). In-hospital mortality was associated with age, lung injury, intensive care unit admission, number of and cardiovascular comorbidities, diabetes, chronic kidney disease, and all inflammatory biomarkers. Serious liver injury at admission (ALT ≥ 5 × ULN) was significantly associated with in-hospital mortality (OR = 7.9, 95%CI 2-28.9). At discharge, drug-induced liver injury (DILI) was found in patients treated with remdesivir, ALT ≥ 2 ULN (OR = 2.62, 95%CI 1.22-5.75). Treatment with dexamethasone, remdesivir, and immunomodulators showed improved survival, OR = 0.50 (95%CI 0.33-0.77). Regardless of the variant and treatment options, less than 2% of patients displayed serious liver injury, which was not found to be a death predictor in multivariable analysis.

The Diagnostic Challenges and Clinical and Serological Outcome in Patients Hospitalized for Suspected Lyme Neuroborreliosis.

The aim of our study was to evaluate the differential diagnosis and clinical/serological outcome to antibiotic treatment in patients hospitalized for suspected Lyme neuroborreliosis (LNB). A prospective study included patients hospitalized in a Romanian hospital between March 2011 and October 2012 with neurological symptoms, positive laboratory tests for Borrelia burgdorferi, cerebrospinal fluid (CSF) analysis, and no previous treatment for LNB. A questionnaire was completed for each patient at admission, at the end of treatment, and 3 months later. Patients were treated with antibiotic therapy (ceftriaxone/cefotaxime), irrespective of CSF analysis results. A symptomatic scoring scale was used for the follow-up. Out of the 42 patients included, no patient fulfilled criteria for definite LNB; 7 patients were classified as possible LNB; and in 33 patients, LNB was excluded. Two patients could not be classified (insufficient amount of CSF). Clinical follow-up suggested a better response to therapy in the group of patients with possible LNB than in the group with LNB excluded. The patients' differential diagnosis and serological follow-up are presented. Patients investigated for suspected LNB present diverse clinical manifestations and comorbidities that complicate differential diagnosis. LNB may be misdiagnosed if CSF analysis is not performed.

External Validation of Mortality Scores among High-Risk COVID-19 Patients: A Romanian Retrospective Study in the First Pandemic Year.

BACKGROUND: We aimed to externally validate three prognostic scores for COVID-19: the 4C Mortality Score (4CM Score), the COVID-GRAM Critical Illness Risk Score (COVID-GRAM), and COVIDAnalytics. METHODS: We evaluated the scores in a retrospective study on adult patients hospitalized with severe/critical COVID-19 (1 March 2020-1 March 2021), in the Teaching Hospital of Infectious Diseases, Cluj-Napoca, Romania. We assessed all the deceased patients matched with two survivors by age, gender, and at least two comorbidities. The areas under the receiver-operating characteristic curves (AUROCs) were computed for in-hospital mortality. RESULTS: Among 780 severe/critical COVID-19 patients, 178 (22.8%) died. We included 474 patients according to the case definition (158 deceased/316 survivors). The median age was 75 years; diabetes mellitus, malignancies, chronic pulmonary diseases, and chronic kidney and moderate/severe liver diseases were associated with higher risks of death. According to the predefined 4CM Score, the mortality rates were 0% (low), 13% (intermediate), 27% (high), and 61% (very high). The AUROC for the 4CM Score was 0.72 (95% CI: 0.67-0.77) for in-hospital mortality, close to COVID-GRAM, with slightly greater discriminatory ability for COVIDAnalytics: 0.76 (95% CI: 0.71-0.80). CONCLUSION: All the prognostic scores showed close values compared to their validation cohorts, were fairly accurate in predicting mortality, and can be used to prioritize care and resources.

Seroprevalence Rates against West Nile, Usutu, and Tick-Borne Encephalitis Viruses in Blood-Donors from North-Western Romania.

INTRODUCTION: West Nile virus (WNV), Usutu virus (USUV), and the tick-borne encephalitis virus (TBEV) are all arboviruses belonging to Flaviviridae family. All are characterized by vectorial transmission and sometimes associated with neuroinvasive infections. The circulation of these viruses is considered endemic in parts of Europe, with human cases reported in many countries. Among hosts, the viruses are vectored by hematophagous arthropods, such as mosquitoes (WNV, USUV) and ticks (TBEV). Considering the currently outdated knowledge regarding the epidemiology of these viruses in Romania, the aim of our study was to assess the seroprevalence rates of WNV, USUV, and TBEV among healthy blood donors in north-western Romania. METHODS: Human blood samples from healthy donors were collected between November 2019 and February 2020 in six counties from the north-western region of Romania. The samples were serologically tested by ELISA and serum neutralization test. RESULTS: Overall, we obtained a seroprevalence of 3.17% for WNV, 0.08% for TBEV, and 0% for USUV. CONCLUSION: Despite the low seroprevalence of WNV, USUV, and TBEV in our study, we highlight the need for continuous nationwide vector and disease surveillance and implementation of control measures. Further research is required for an optimal overview of the epidemiological status of the Romanian population regarding these flaviviruses together with countrywide awareness campaigns.

Longitudinal determination of BNT162b2 vaccine induced strongly binding SARS-CoV-2 IgG antibodies in a cohort of Romanian healthcare workers.

Mass vaccination against the disease caused by the novel coronavirus (COVID-19) was a crucial step in slowing the spread of SARS-CoV-2 in 2021. Even in the face of new variants, it still remains extremely important for reducing hospitalizations and COVID-19 deaths. In order to better understand the short- and long-term dynamics of humoral immune response, we present a longitudinal analysis of post-vaccination IgG levels in a cohort of 166 Romanian healthcare workers vaccinated with BNT162b2 with weekly follow-up until 35 days past the first dose and monthly follow-up up to 6 months post-vaccination. A subset of the patients continued with follow-up after 6 months and either received a booster dose or got infected during the Delta wave in Romania. Tests were carried out on 1694 samples using a CE-marked IgG ELISA assay developed in-house, containing S1 and N antigens of the wild type virus. Participants infected with SARS-CoV-2 before vaccination mount a quick immune response, reaching peak IgG levels two weeks after the first dose, while IgG levels of previously uninfected participants mount gradually, increasing abruptly after the second dose. Overall higher IgG levels are maintained for the previously infected group throughout the six month primary observation period (e.g. 36-65 days after the first dose, the median value in the previously infected group is 5.29 AU/ml, versus 3.58 AU/ml in the infection naïve group, p less than 0.001). The decrease of IgG levels is gradual, with lower median values in the infection naïve cohort even 7-8 months after vaccination, compared to the previously infected cohort (0.7 AU/ml versus 1.29 AU/ml, p = 0.006). Administration of a booster dose yielded higher median IgG antibody levels than post second dose in the infection naïve group and comparable levels in the previously infected group.