Measles Immunity Status of Greek Population after the Outbreak in 2017-2018: Results from a Seroprevalence National Survey.

Accurate data on susceptibility rates against measles in the general population of Greece are scarce. Many studies have estimated the vaccination coverage, but none have calculated the nationwide immunity rate, including all age groups, against the measles virus. The purpose of our study was to determine the measles immunity status, especially after the latest outbreak in 2017-2018. In total, 3972 leftover blood samples were obtained during 2020-2021. They were collected from a nationwide laboratory network using a geographically stratified sampling strategy and were tested for the presence of measles-specific IgG antibodies. The overall crude seroprevalence was calculated to be 89.6% and the adjusted was 89.8% (95% CI: 88.8-90.8%). There was no statistically significant difference in seropositivity between sexes (p = 0.783). Higher immunity rates and antibody concentrations were found in older age groups ≥41 years old (94.9%, 95% CI: 93.7-95.9%, and 730.0 mIU/mL) in comparison with younger individuals aged 1-40 years old (83.4%, 95% CI: 81.6-85.7%, and 616.5 mIU/mL). Comparing the seroprevalence among the Nomenclature of Territorial Units for Statistics (NUTS 2), a statistically significant difference was estimated among them (<0.001). The two regions where higher measles incidence was observed during the 2017-2018 outbreak, Eastern Macedonia and Thrace, and Western Greece, were among the four regions with lower seropositivity (84.6%, 95% CI: 79.9-89.4%, and 85.9%, 95% CI: 81.4-90.4%, respectively). Our study showed a measles immunity gap that affects the younger age groups and makes a new measles outbreak likely. The enforcement of vaccination campaigns and addressing vaccine hesitancy could bridge it and achieve the required target of herd immunity.

A Point Source Gastroenteritis Outbreak in a High School Putatively Due to Clostridium perfringens: Timely Investigation Is Everything.

On May 28, 2021, several gastroenteritis cases were reported among students and staff of a high school on a Greek island. Lunches were delivered daily at the school by a catering company. We conducted a retrospective cohort study. Cases were defined as students or school staff presenting with diarrhea and abdominal pain between May 27 and 29, 2021. We used a self-administered questionnaire to collect the data. In the univariate analysis, we calculated food-specific attack rates (ARs), relative risks (RRs), and confidence intervals (95% CIs). Statistically significant variables (p < 0.05) were included in the multivariable model. Stool samples were collected from students on the same day of notification and were tested for bacteria, viruses, and parasites. The local food safety authority also inspected the school and the catering company premises. The food samples were sent to the reference laboratory for analysis. We received 129 responses from 174 students and staff members (response rate: 74%). Thirty cases were identified (AR, 23%). Of the 11 significant food items in the univariate analysis, the consumption of spaghetti with minced meat on May 27 (odds ratio 53.40, 95% CI: 3.9-736.1, 97% of cases exposed) was independently associated with the onset of gastroenteritis. Clostridium perfringens was detected in two of the three students' stool samples. Six samples from spaghetti with minced meat were positive for C. perfringens and four of those also tested positive for Bacillus cereus. Environmental investigation revealed non-compliances concerning food personnel training and Hazard Analysis and Critical Control Points (HACCP) plan implementation in the food establishment. Timely investigation and samples collection allowed the identification of the probable source of the outbreak. Changes, regarding the reception and distribution of meals in the school, were applied. Recommendations were given to the catering establishment focusing on adequate personnel training and HACCP issues, especially regarding temperature/time controls during all production, handling, and distribution phases.

Does tourism affect the long term course of COVID-19 pandemic in a country of destination? Evidence from a popular Greek island in 2020 where control measures were implemented.

Greece opened its points of entry on July 1, 2020, with specific guidelines for travellers arriving by sea, air or land. The aim of this article is to examine the effect of tourism on the long term course of the Coronavirus Disease 2019 (COVID-19) pandemic during the pre-vaccination era (June to December 2020) on the popular Greek island of Crete. To achieve this, a cross-sectional serosurvey, repeated at monthly intervals, was conducted to compare the seroprevalence in Crete with seroprevalence in the mainland of Greece. Crete welcomed nearly 2,000,000 travellers during the 2020 summer season. Left-over serum samples were collected and obtained from public and private laboratories located in Greece, including the island of Crete. These samples were tested for the presence of anti-SARS-CoV-2 IgG antibodies. A total of 55,938 samples were collected, 3,785 of which originated from Crete. In Crete, the seroprevalence ranged between 0% (June 2020) and 2.58% (December 2020), while the corresponding seroprevalence in Greece was 0.19% and 10.75%, respectively. We identified 4.16 times lower seropositivity in Crete (2.58%) in comparison with the mainland of Greece (10.75%) during December 2020. Moreover, the monthly infection fatality rate (IFR) in Crete was calculated at 0.09%, compared with 0.21% in mainland Greece for December 2020. The island of Crete presented more than four times lower seroprevalence than the mainland of Greece, despite being a highly attractive tourist destination. This evidence supports the idea that tourism may not have affected the long term course of the COVID-19 pandemic in Greece. However, due to contradicting results from previous studies, further investigation is needed.

Dynamics of Anti-SARS-CoV-2 IgA and IgG Responses and Their Protective Effect against Fatal Disease after Booster COVID-19 Vaccination.

During the post-coronavirus disease (COVID-19) era, a primary question is whether booster vaccination is effective against severe COVID-19 and should be recommended, particularly to individuals at high risk for severe disease (i.e., the elderly or those with additional severe comorbidities). From December 2020 to February 2023, a cohort study was conducted to estimate IgG and IgA immunogenicity and the dynamics of booster mono- and bivalent COVID-19 mRNA vaccines in 260 individuals (male/female: 114/146, median age: 68 years, interquartile range (IQR) = 31) who initially received either mRNA (218) or adenovirus-vector-based vaccines (42). Participants were followed until the 90th day after the third booster dose. Our cohort study indicated a beneficial effect of booster vaccination on the magnitude of IgG and IgA severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) antibodies. We found that second and third booster doses were more protective than one against fatal disease (p = 0.031, OR 0.08). In conclusion, booster COVID-19 vaccination should be strongly recommended, especially to individuals at high risk for severe/fatal disease.

Intensity of Humoral Immune Responses, Adverse Reactions, and Post-Vaccination Morbidity after Adenovirus Vector-Based and mRNA Anti-COVID-19 Vaccines.

The aim of the study was to compare mRNA vaccine BNT162b2 with adenovirus vector- based vaccines in terms of presence of adverse reactions, immunogenicity, and protection against COVID-19. A total of 270 individuals were enrolled, of which 135 were vaccinated with adenovirus vector-based vaccines and compared with 135 age- and sex-matched participants who received the BNT162b2 mRNA vaccine. Serum sampling was performed on all participants on days 21, 42, 90, and 180 following the first dose, to evaluate anti-spike IgG and IgA responses. Antibodies were quantified by chemiluminescent microplate and ELISA assays. We demonstrate that both mRNA and adenovirus vector-based vaccines caused mild side-effects and were effective in inducing adequate antibody responses against SARS-CoV-2, although BNT162b2 was superior concerning the intensity of antibody responses and protection against severe COVID-19. Moreover, we identify that IgG and IgA responses depended primarily on both history of previous COVID-19 infection and vaccination platform used, with individuals immunized with a single-dose vaccine having lower antibody titers over time. Lastly, all vaccine platforms had limited side-effects, with the most frequent pain at the injection site. Our results provide useful information regarding antibody responses after vaccination with different vaccine platforms, which can be useful for public health vaccination strategies.

Performance Evaluation of a Rapid Antigen Test (RAT) during Omicron Pandemic Wave in Greece, Conducted by Different Personnel, and Comparison with Performance in Previous Wave (Alpha Variant) Period.

Due to the prevailing ambiguity regarding the performance of rapid antigen tests (RATs) for B.1.1.529 (Omicron) variant diagnosis, a commercial RAT was evaluated in the emergency ward of a general hospital in Larissa, Central Greece. The sampling and the evaluation were repeated twice by different personnel. Discordance between the two samplings was observed regarding the sensitivity (47.5%, 95% CI: 39.0-56.1 vs. 78.6%, 95% CI: 69.1-86.2) and specificity (93.8%, 95% CI: 86.0-97.9 vs. 100.0%, 95% CI: 93.3-100.0) of the RAT. Furthermore, the test displayed slightly lower sensitivity (78.6% vs. 85.5%, 95% CI: 79.1-90.5) compared to its initial evaluation that was conducted by our team when the B.1.1.7 (Alpha) variant was dominant.

SARS-CoV-2 Sero-Surveillance in Greece: Evolution over Time and Epidemiological Attributes during the Pre-Vaccination Pandemic Era.

BACKGROUND: Nation-wide SARS-CoV-2 seroprevalence surveys provide valuable insights into the course of the pandemic, including information often not captured by routine surveillance of reported cases. METHODS: A serosurvey of IgG antibodies against SARS-CoV-2 was conducted in Greece between March and December 2020. It was designed as a cross-sectional survey repeated at monthly intervals. The leftover sampling methodology was used and a geographically stratified sampling plan was applied. RESULTS: Of 55,947 serum samples collected, 705 (1.26%) were found positive for anti-SARS-CoV-2 antibodies, with higher seroprevalence (9.09%) observed in December 2020. Highest seropositivity levels were observed in the "0-29" and "30-49" year age groups. Seroprevalence increased with age in the "0-29" age group. Highly populated metropolitan areas were characterized with elevated seroprevalence levels (11.92% in Attica, 12.76% in Thessaloniki) compared to the rest of the country (5.90%). The infection fatality rate (IFR) was estimated at 0.451% (95% CI: 0.382-0.549%) using aggregate data until December 2020, and the ratio of actual to reported cases was 9.59 (7.88-11.33). CONCLUSIONS: The evolution of seroprevalence estimates aligned with the course of the pandemic and varied widely by region and age group. Young and middle-aged adults appeared to be drivers of the pandemic during a severe epidemic wave under strict policy measures.

Intensity and Dynamics of Anti-SARS-CoV-2 Immune Responses after BNT162b2 mRNA Vaccination: Implications for Public Health Vaccination Strategies.

The aim of our study was to investigate the immunogenicity of the BNT162b2 vaccination according to the age and medical status of vaccinated individuals. A total of 511 individuals were enrolled (median age: 54.0 years, range: 19-105); 509 of these individuals (99.6%) received two doses of BNT162b2 at an interval of 21 days. IgG and IgA responses were evaluated on days 21, 42, 90, and 180 after the first dose with chemiluminescent microparticle and ELISA assays. The cell-mediated immune responses were assessed by an automated interferon-gamma release assay. We demonstrated positive antibody responses after vaccination for the majority of enrolled participants, although waning of IgG and IgA titers was also observed over time. We further observed that the intensity of humoral responses was positively correlated with increased age and prior COVID-19 infection (either before or after the first vaccination). Moreover, we found that only a medical history of autoimmune disease could affect the intensity of IgA and IgG responses (3 weeks after the primary and secondary immunization, respectively), while development of systemic adverse reactions after the second vaccination dose was significantly associated with the height of IgG responses. Finally, we identified a clear correlation between humoral and cellular responses, suggesting that the study of cellular responses is not required as a routine laboratory test after vaccination. Our results provide useful information about the immunogenicity of COVID-19 vaccination with significant implications for public health vaccination strategies.

Short-Term Selection to Diflubenzuron and Bacillus thuringiensis Var. Israelensis Differentially Affects the Winter Survival of Culex pipiens f. Pipiens and Culex pipiens f. Molestus (Diptera: Culicidae).

The Culex pipiens (Diptera: Culicidae) mosquito is of high medical importance as it is considered the prime vector of West Nile virus. In Europe, this species consists of two forms, named pipiens and molestus, that exhibit substantial differences in their overwintering biology. Diflubenzuron (DFB) and Bacillus thuringiensis var. israelensis (Bti) are two of the most used larvicides in mosquito control, including that of Culex pipiens. The high dependency on these two larvicides poses major concerns for resistance development. The evolution and stability of resistance to insecticides has been associated with fitness costs that may be manifested under stressful conditions, such as the winter period. This study investigated how short-term selection of pipiens and molestus forms to both larvicides affect their overwintering success. Larvae from each form were subjected to the same selective pressure (80% mortality) for three successive generations with DFB and Bti. At the end of this process, the winter survival between the selected populations and the controls (colonies without selection) was determined for each form. Selection to both larvicides significantly reduced the winter survival rates of molestus but not of pipiens form, indicating potential differences in the persistence of the selected individuals from year to year between the two forms.

Repeated Leftover Serosurvey of SARS-CoV-2 IgG Antibodies in Greece, May to August 2020.

A serosurvey of IgG antibodies against SARS-CoV-2 was conducted in Greece between May and August 2020. It was designed as a cross-sectional survey and was repeated at monthly intervals. The leftover sampling methodology was used and a geographically stratified sampling plan was applied. Of 20,110 serum samples collected, 89 (0.44%) were found to be positive for anti-SARS-CoV-2 antibodies, with higher seroprevalence (0.35%) observed in May 2020. The highest seroprevalence was primarily observed in the "30-49" year age group. Females presented higher seroprevalence compared to males in May 2020 (females: 0.58% VS males: 0.10%). This difference reversed during the study period and males presented a higher proportion in August 2020 (females: 0.12% VS males: 0.58%). Differences in the rate of seropositivity between urban areas and the rest of the country were also observed during the study period. The four-month infection fatality rate (IFR) was estimated to be 0.47%, while the respective case fatality rate (CFR) was at 1.89%. Our findings confirm low seroprevalence of COVID-19 in Greece during the study period. The young adults are presented as the most affected age group. The loss of the cumulative effect of seropositivity in a proportion of previous SARS-CoV-2 infections was indicated.

Transmission Dynamics of SARS-CoV-2 during an Outbreak in a Roma Community in Thessaly, Greece-Control Measures and Lessons Learned.

A COVID-19 outbreak occurred among residents of a Roma settlement in Greece (8 April-4 June 2020). The aim of this study was to identify factors associated with an increased risk of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection and to evaluate the effectiveness of control measures implemented. Data were analyzed from individuals that were tested for SARS-CoV-2 during contact tracing, population screening or hospital visits. RT-PCR was used for the detection of SARS-CoV-2 in oropharyngeal samples. Risk factors for household secondary attack rates (SAR) and hospitalization with COVID-19 were examined using chi-square tests, Fisher's exact tests and logistic regression analyses. During the outbreak, 142 cases, 20 hospitalizations and 1 death were recorded, with a total of 2273 individuals tested. The risk of hospitalization was associated with age (OR: 1.04, 95% CI: 1.02-1.07) and Cycle threshold (Ct) values (OR for a decrease in Ct values by 1: 1.18, 95% CI: 1.07-1.31). Household SAR was estimated at 38.62% (95% CI: 32.50-45.01%). After the designation of an isolation facility for cases, household SAR declined from 74.42% to 31.03%. Household size was associated with the risk of infection (OR: 2.65, 95% CI: 1.00-7.07). The presence of COVID-19 symptoms among index cases was correlated with higher transmission (OR: 23.68, 95% CI 2.21-253.74) in multivariate analysis, while age was found to be associated with SAR only in univariate analysis. Roma communities can be particularly vulnerable to the spread of SARS-CoV-2. In similar settings, symptomatic cases are more important transmitters of SARS-CoV-2. Within these communities, immediate measures should be implemented to mitigate disease spread.

Repeated leftover serosurvey of SARS-CoV-2 IgG antibodies, Greece, March and April 2020.

A serosurvey of IgG antibodies against severe acute respiratory coronavirus 2 (SARS-CoV-2) was performed during March and April 2020. Among 6,586 leftover sera, 24 (0.36%) were positive, with higher prevalence in females, older individuals and residents of large urban areas. Seroprevalence was estimated at 0.02% and 0.25%, respectively, in March and April, infection fatality rate at 2.66% and 0.54%. Our findings confirm low COVID-19 incidence in Greece and possibly the effectiveness of early measures.

Serotyping and detection of pathogenecity loci of environmental isolates of Legionella pneumophila using MALDI-TOF MS.

BACKGROUND: The majority of Legionnaires' disease cases is attributed to Legionella pneumophila serogroup 1 (Lp1). Moreover, pathogenicity loci lvh and rtxA were associated with the ability of Lp strains to cause the disease. Consequently, except from serogroup assignment the detection of the aforementioned virulence genes during Legionella detection in water samples, could help environmental risk assessment and the implementation of targeted control measures. AIM: To establish and validate a rapid and robust MALDI-TOF MS-based method for the assignment of Lp isolates to serogroup, and identify distinct peak biomarkers for the detection of lvh and rtxA loci during environmental investigations. METHOD: Fifteen reference strains and 150 Lp environmental isolates (70 Lp1 and 80 Lp2-15 strains) were used. All strains were PCR-tested for the presence of lvh and rtxA loci. Independent training and validation strain sets were constituted and all strains were protein-extracted and submitted to MALDI-TOF MS analysis. The raw spectra of the training set strains obtained, were introduced into the Mass-Up software platform for biomarker detection, for both serogroup assignment and pathogenicity loci detection. Validation of the assigned biomarkers followed using the validation set strains. RESULTS: For serogroup assignment, the Mass-up analysis indicated five potential discriminating peaks and correctly classified 115 out of 132 validation set strains, displaying sensitivity of 87.5%, specificity of 86.7% and 87.1% accuracy. Concerning the pathogenicity loci detection, the Mass-up analysis indicated two ion peaks for rtxA locus discrimination and one peak for lvh locus discrimination. Concerning the lvh virulence gene, the algorithm correctly classified 113 out of 137 positive and all negative strains 14 in total-showing sensitivity of 82.5%, specificity of 100.% and 84.1% accuracy. For rtxA locus, 134 out of 134 positive and 14 out of 17 negative strains were correctly classified with sensitivity of 100%, specificity of 76.5% and 97.4% accuracy. CONCLUSION: MALDI-TOF MS displayed good performance for Lp serogroup assignment and detection of the lvh and rtxA virulence genes. These findings could contribute to the rapid, inexpensive and comprehensive case investigation and risk assessment. Further studies are needed to standardize and evaluate the method using the direct target plate protein profiling instead of protein extraction in order to simplify the protocol. .

Evaluation of Two Chemiluminescent and Three ELISA Immunoassays for the Detection of SARS-CoV-2 IgG Antibodies: Implications for Disease Diagnosis and Patients' Management.

The estimation of anti-SARS-CoV-2 IgG antibodies is possibly the best approach to accurately establish the number of infected individuals and the seroprevalence of COVID-19 within a population. Thus, several commercial immunoassays have recently been developed. The purpose of our study was to assess the performance of five commonly used immunoassays in Greece (3 ELISA, namely Euroimmun SARS-CoV-2, GA GENERIC SARS-CoV-2 and Vircell COVID-19; and 2 chemiluminescent, namely ABBOTT SARS-CoV-2 and ROCHE Elecsys Anti-SARS-CoV-2 test) for the detection of anti-SARS-CoV-2 IgG antibodies. Sera specimens derived from 168 individuals were utilized to assess the specificity and sensitivity score of each assay. Among them, we included 99 COVID-19 patients (29 asymptomatic, 36 with symptom onset 4 to 14 days before serum sampling, and 34 with symptom initiation ≥ 15 days ago), and 69 volunteers with sera specimens collected prior to the SARS-CoV-2 outbreak and maintained at -80°C. We demonstrated that chemiluminescent immunoassays exhibit a significantly higher specificity score but a lower sensitivity, compared to ELISA immunoassays. Moreover, immunoassays detecting IgG antibodies against SARS-CoV-2 N protein instead of S protein alone are more reliable, considering both specificity and sensitivity scores. Interestingly, all asymptomatic patients displayed anti-SARS-CoV-2 IgG antibodies, confirmed by at least two immunoassays. We suggest that chemiluminescent assays could be used as screening methods for the detection of anti-SARS-CoV-2 antibodies to evaluate the possible prevalence of disease in the general population, while ELISA assays would be more reliable to evaluate, and follow-up confirmed COVID-19 patients.

Legionella Colonization of Hotel Water Systems in Touristic Places of Greece: Association with System Characteristics and Physicochemical Parameters.

This study aimed to assess the colonization of hotel water systems in central Greece and Corfu by Legionella, and to investigate the association between physicochemical parameters and Legionella colonization. Standardized hygiene inspection was conducted in 51 hotels, and 556 water samples were analyzed for Legionella spp. Free chlorine concentration, pH, hardness, conductivity, and trace metals were defined in cold water samples. The results of inspections and chemical analyses were associated with the microbiological results using univariate and logistic regression analysis. According to the score of the checklist used for the inspections, 17.6% of the hotels were classified as satisfactory, 15.7% as adequate, and 66.7% as unsatisfactory. Moreover, 74.5% of the hotels were colonized by Legionella spp. and 31.4% required remedial measures according to the European guidelines. Legionella spp. were isolated in 28% of the samples. Unsatisfactory results of inspections were associated with Legionella presence (relative risk (RR) = 7.67, p-value = 0.043). In hot-water systems, <50 °C temperatures increased the risk of Legionella colonization (RR = 5.36, p-value < 0.001). In cold-water systems, free chlorine concentration <0.375 mg/L (odds ratio (OR) = 9.76, p-value = 0.001), pH ≥ 7.45 (OR = 4.05, p-value = 0.007), and hardness ≥321 mgCaCO3/L (OR = 5.63, p-value = 0.003) increased the risk, whereas copper pipes demonstrated a protective role (OR = 0.29, p-value = 0.0024). The majority of the hotels inspected were colonized with Legionella. Supplementary monitoring of the risk factors that were identified should be considered.

Clusters of Healthcare-Associated Legionnaires' Disease in Two Hospitals of Central Greece.

Healthcare-associated Legionnaires' disease often leads to fatal respiratory tract infection among hospitalized patients. In this report, three cases of Legionnaires' disease among patients in two different hospitals (Hospital A and Hospital B) were investigated. After conducting an epidemiologic and environmental investigation, the water distribution systems (WDSs) were identified as the possible source of infection, as Legionella pneumophila serogroup 1 (Lp1) was isolated from both clinical and environmental samples. Patients received aerosol therapy with nebulizers during their hospitalization. Based on the results of the investigation, the hospitals' infection control committees reviewed their policies for Legionnaires' disease prevention and implemented control measures focusing on using sterile fluids for aerosol treatments.

First reported isolation of an emerging opportunistic pathogen (Elizabethkingia anophelis) from hospital water systems in Greece.

After the Elizabethkingia anophelis outbreak in Wisconsin, USA, an active search for the detection of the microorganism in hospital water systems from Central and Northern Greece was performed from June to December 2016. In total, 457 water samples from 11 hospitals were analyzed. Elizabethkingia spp. was detected in three samples collected from two hospitals, both of which are located in Northern Greece. Two of the three isolated strains were identified as Elizabethkingia anophelis. No cases of Elizabethkingia infection were reported in either hospital during 2016. This is the first reported isolation of the pathogen in water supply systems in Greece.