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.

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.

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.

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.

A pilot sentinel surveillance system to monitor treatment and treatment outcomes of chronic hepatitis B and C infections in clinical centres in three European countries, 2019.

BackgroundThe World Health Organization European Action Plan 2020 targets for the elimination of viral hepatitis are that > 75% of eligible individuals with chronic hepatitis B (HBV) or hepatitis C (HCV) are treated, of whom > 90% achieve viral suppression.AimTo report the results from a pilot sentinel surveillance to monitor chronic HBV and HCV treatment uptake and outcomes in 2019.MethodsWe undertook retrospective enhanced data collection on patients with a confirmed chronic HBV or HCV infection presenting at one of seven clinics in three countries (Croatia, Romania and Spain) for the first time between 1 January 2019 and 30 June 2019. Clinical records were reviewed from date of first attendance to 31 December 2019 and data on sociodemographics, clinical history, laboratory results, treatment and treatment outcomes were collected. Treatment eligibility, uptake and case outcome were assessed.ResultsOf 229 individuals with chronic HBV infection, treatment status was reported for 203 (89%). Of the 80 individuals reported as eligible for treatment, 51% (41/80) were treated of whom 89% (33/37) had achieved viral suppression. Of 240 individuals with chronic HCV infection, treatment status was reported for 231 (96%). Of 231 eligible individuals, 77% (179/231) were treated, the majority of whom had received direct acting antivirals (99%, 174/176) and had achieved sustained virological response (98%, 165/169).ConclusionTreatment targets for global elimination were missed for HBV but not for HCV. A wider European implementation of sentinel surveillance with a representative sample of sites could help monitor progress towards achieving hepatitis control targets.

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.

Seroprevalence of antibodies against Borrelia burgdorferi sensu lato in healthy blood donors in Romania: an update.

BACKGROUND: The Borrelia burgdorferi sensu lato (s.l.) genogroup is the causative agent responsible for Lyme borreliosis, a common tick-borne infectious disease in some temperate regions of the Northern Hemisphere. In humans, the clinical manifestations of Lyme borreliosis vary from dermatological infection to severe systemic manifestations. In Romania, data on the seroprevalence of Lyme borreliosis and associated risk factors are scarce and outdated, as the only seroprevalence study with a large dataset was published more than 20 years ago. Therefore, the aim of the present study was to evaluate the seroprevalence for Borrelia burgdorferi s.l. in healthy blood donors from six Romanian counties and identify the associated risk factors. METHODS: The study was conducted among 1200 healthy blood donors aged between 18 and 65 years during November 2019 and September 2020 from six counties in the northwestern and central parts of Romania. A two-tiered testing strategy was applied. Positive and equivocal immunoenzymatic test results for IgG and IgM antibodies were further confirmed by Western blot. RESULTS: Serum samples from 20% of the blood donors had positive or equivocal IgG and IgM ELISA index values. In total, 2.3% of the serum samples for IgG and 1.8% for IgM were positive by Western blot. The seroprevalence for both antibodies varied between 1.5% (Satu-Mare) and 6.5% (Bistrița-Nasaud) in the six counties investigated. The highest seroprevalence was observed in men (4.7%), in blood donors performing their professional activities outdoors (4.2%), and in those aged ≥ 56 years (8%). CONCLUSIONS: These findings confirm the presence of specific IgG and IgM antibodies to B. burgdorferi s.l. among healthy blood donors from Romania. Furthermore, potential risk factors, such as gender, age, and behavior, associated with the presence of positive B. burgdorferi s.l. antibodies among healthy blood donors were identified.

Evaluation of several serum interleukins as markers for treatment effectiveness in naïve HIV infected patients: A pilot study.

In this observational pilot study, we investigated the impact of Dolutegravir, Raltegravir, Elvitegravir (Integrase Strand Transfer Inhibitors, INSTIs), or boosted Darunavir (a Protease Inhibitor, PI) in combination with two nucleoside reverstranscriptase inhibitors (Emtricitabine/Tenofovir disoproxil or Lamivudine/Tenofovir disoproxil, NRTI) on four interleukins (IL-4, IL-10, IL-13, and IL-21) as immune activation markers in naïve HIV(Human Immunodeficiency Virus)-infected patients during the first six months of combined standard-of-care antiretroviral therapy (cART). Newly diagnosed with HIV-infected subjects and without any disease that could affect the immune activation markers were evaluated. The patients' physicians recommended the cART as standard-of-care and the ILs were measured before cART and six months of cART. The levels of CD4+ T-cells count and CD4+/CD8+ ratio significantly increased at six months (P-value<0.02) regardless of the drugs, INSTIs or PI. However, a CD4+/CD8+ >1 was observed in 25% of patients treated with Raltegravir and half of those treated with Dolutegravir. At six months of cART, viral load was detectable in only 6/31 individuals. IL-21 had an undetectable level in 30/31 patients after six months of cART. Our results suggest the potency in restoring immune markers in HIV-infected patients with all investigated drugs. Dolutegravir showed a tendency to statistically significant changes in IL-4 and IL-10. A clinical trial with random allocation of medication and an extensive follow-up is needed to replicate this research and validate the usefulness of evaluated ILs as markers of cART effectiveness.

Continuum of Care UNAIDS Fast-Track Targets Evaluation of Patients Living with Human Immunodeficiency Virus Infection.

The current study evaluated the progress of continuum healthcare for patients living with human immunodeficiency virus (HIV) infection from Cluj County in two moments, 2016 and 2020, and compared the results to the Fast-Track targets (FTTs) proposed by the Joint United Nations Programme (UNAIDS) on HIV/AIDS. By the end of 2020, 368 out of 385 confirmed HIV-positive patients from Cluj County were under surveillance in our center, representing almost 95% of the patients living with HIV and knowing their diagnosis, compared to 87.9% in 2016. Nearly 97% of those in active follow-up from Cluj County were under antiretroviral therapy (ART) in 2020, compared to 89% in 2016. The number of virally suppressed patients from those under ART was almost 94% in 2020, compared to 82.7% in 2016, and the increase is observed regardless of the ART regime. A shift towards integrase strand transfer inhibitors, with a higher efficacy, fewer adverse effects, and fewer drug interactions, is observed, which could contribute to the decrease in HIV transmission.

A View of Human Immunodeficiency Virus Infections in the North-West Region of Romania.

Background and objectives: In Romania, the human immunodeficiency virus (HIV) epidemic is almost the same as it is in Central Europe, with some differences; particularity the following one: people with nosocomial HIV infection, also known as Romanian cohort. Aim: The study aimed to present a local view of HIV infection in the North-West part of Romania, and to identify the particularities of patients under medical care in the Cluj AIDS Center. Materials and Methods: The demographic characteristics (age and gender), and medical and epidemiological data (stage of HIV infection and mode of transmission) of patients in a medical care in the Cluj Acquired Immunodeficiency Syndrome (AIDS) Center were evaluated. Data from the first patients admitted between 1989 and 2018, and the statuses of the infected persons as per 31 December 2018 were analyzed. Results: Nine hundred and fourteen patients were included in the study. The patients' ages varied from 0 (newborns from HIV-infected mothers) to 72 years old, and most patients were men (596 men vs. 318 women). The main mode of transmission was sexual (>50%), with an increased number of men who have sex with men (MSM) in the last years (from two cases in 2006 to thirty-four cases in 2018), and a very small percentage of intravenous drug users (IDU; <1%). The patients from the Romanian cohort were more frequently women as compared with men (p-value <0.0001), women were more frequently later presenters than men (p-value <0.0001), and the women more frequently had candidosis (p-value = 0.0372), cerebral toxoplasmosis (p-value = 0.0404), and co-infection with hepatitis B virus (p-value = 0.0018). One hundred and sixty patients died by the end of 2018 (17.5%). Sixty-eight children had been born from HIV-infected mothers, and 17 were HIV infected (25%). Conclusion: The main mode of HIV transmission in our sample was sexual, with an increased number of MSM over the last years and a low number of cases of intravenous drug users. A quarter of children borne from HIV-infected mothers were HIV infected.

Assesment of the Duke criteria for the diagnosis of infective endocarditis after twenty-years. An analysis of 241 cases.

BACKGROUND AND AIMS: In the absence of classical features (fever, cardiac murmur, and peripheral vascular stigmata) the diagnosis of infective endocarditis (IE) may be difficult. Current clinical guidelines for the diagnosis and management of IE recommend the use of modified Duke criteria. Correct and prompt diagnosis of IE is crucial for the treatment and outcome of the patients. The aim of this study was to evaluate the presence and the individual value of each criterion of the modified Duke criteria in our patients with infective endocarditis. METHODS: We performed a prospective observational study between January 2008 - June 2014, in which we enrolled consecutive adult patients admitted for suspicion of IE to the Hospital of Infectious Diseases and at the Heart Institute . We used and extensive database in order to collect demographic data, laboratory and echocardiography results, evolution and outcome of the patients. Using the modified Duke criteria we identified 3 categories of IE: definite, possible and rejected. In order to evaluate the importance of each criterion in the diagnosis of IE we tested two hypotheses. First, we excluded each criterion from the final diagnosis and we counted how many cases felt into a lower category. Second, after adding each major and minor criterion, we tested how many cases would have been classifiable as definite IE. RESULTS: The study included 241 adult patients with a mean age 58.16 years and sex ratio male/female 1.94. According to the modified Duke criteria 137 patients had definite IE, 79 patients had possible IE and 25 cases had rejected IE We had blood cultures positive IE in 109 cases and blood culture negative IE (BCNE) in 132 (71.21%) cases. Antibiotic treatment prior to blood culture was recorded in 152 (63.07%) patients. In the absence of the echocardiography major criterion, 43% of cases would become possible. After extraction of major microbiological criterion, only one third of definite cases would become possible. Minor criteria such as fever and predisposition contributed to the diagnosis only in 10% of cases. In the presence of vascular or immunological phenomena, or in the presence of minor microbiological criterion, half of the possible IE cases could become possible. CONCLUSION: Twenty-years after their launch, the Duke criteria for the diagnosis of IE continue to be important tools. Low index of suspicion of IE and inappropriate use of antibiotics may have a great negative impact on the diagnosis of IE. Nowadays, the scarcity of classical Osler manifestations - bacteremia, fever and peripheral stigmata - makes the diagnosis of IE a challenge.