Case series study of nosocomial Legionnaires' disease in Apulia region (southern Italy): The role of different molecular methods in identifying the infection source

BACKGROUND AND AIM: Legionnaires' disease is a severe form of pneumonia caused by the inhalation or aspiration of water droplets contaminated with Legionella pneumophila and other Legionella species. These bacteria are commonly found in natural habitats and man-made water systems. Legionnaires' disease is a significant public health problem, especially in healthcare settings where patients may be exposed to contaminated environmental sources. Nosocomial outbreaks have been reported worldwide, leading to high morbidity and mortality rates, and increased healthcare costs. This study aimed to compare, the clonal relationship of clinical L. pneumophila strains from two different hospitals with L. pneumophila strains isolated from the water supply. METHODS: In the period from 2019 to 2021, clinical and environmental strains involved in three cases of legionellosis were compared by means of pulsed field gel electrophoresis and sequence based typing techniques. RESULTS: Our findings highlight the persistence of clonally distinct strains within each hospital examined. Furthermore, the L. pneumophila strains detected from hospital environmental sources were related to the clinical strains isolated, demonstrating the nosocomial origin of these cases. CONCLUSIONS: Therefore, it is important to implement more accurate surveillance systems both for epidemiological studies and to check the effectiveness of remediation procedures. (www.actabiomedica.it).

Preliminary Investigation on Hygienic-Sanitary Quality of Food Vending Machines.

The consumption of hot drinks dispensed from vending machines has become a common practice, both in workplaces and during free time. Every day, millions of bulk drinks are sold, but the quality of the products distributed may not always be guaranteed, as it is related to many factors such as the quality of the water, the raw materials used, and the effectiveness of the equipment's cleaning system. The purpose of this study is to evaluate the hygienic-sanitary requirements of hot drinks and vending machine surfaces. The investigation highlighted the microbial contamination of both coffee and vending machine surfaces. Although the "coffee break" is usually considered a moment of pleasure, apparently not subject to specific jurisdiction, the products dispensed can represent a health risk if the hygienic conditions are not fully respected. Therefore, official controls carried out by the Prevention Department represent a suitable way for evaluating and guaranteeing the hygienic-sanitary requirements, providing for corrective actions, when needed, to protect consumers.

Legionella anisa or Legionella bozemanii? Traditional and molecular techniques as support in the environmental surveillance of a hospital water network.

Understanding the actual distribution of different Legionella species in water networks would help prevent outbreaks. Culture investigations followed by serological agglutination tests, with poly/monovalent antisera, still represent the gold standard for isolation and identification of Legionella strains. However, also MALDI-TOF and mip-gene sequencing are currently used. This study was conducted to genetically correlate strains of Legionella non pneumophila (L-np) isolated during environmental surveillance comparing different molecular techniques. Overall, 346 water samples were collected from the water system of four pavilions located in a hospital of the Apulia Region of Italy. Strains isolated from the samples were then identified by serological tests, MALDI-TOF, and mip-gene sequencing. Overall, 24.9% of water samples were positive for Legionella, among which the majority were Legionella pneumophila (Lpn) 1 (52.3%), followed by Lpn2-15 (20.9%), L-np (17.4%), Lpn1 + Lpn2-15 (7.1%), and L-np + Lpn1 (2.3%). Initially, L-np strains were identified as L. bozemanii by monovalent antiserum, while MALDI-TOF and mip-gene sequencing assigned them to L. anisa. More cold water than hot water samples were contaminated by L. anisa (p < 0.001). PFGE, RAPD, Rep-PCR, and SAU-PCR were performed to correlate L. anisa strains. Eleven out of 14 strains identified in all four pavilions showed 100% of similarity upon PFGE analysis. RAPD, Rep-PCR, and SAU-PCR showed greater discriminative power than PFGE.

Vaccination Offer during the Occupational Health Surveillance Program for Healthcare Workers and Suitability to Work: An Italian Retrospective Cohort Study.

The active immunization of health care workers (HCWs) is a crucial measure to avoid nosocomial infection; nevertheless, vaccine coverage (VC) among health personnel in Italy is unsatisfactory. To improve VC in the healthcare set, the Hygiene and Occupational Medicine departments of Bari Policlinico General University Hospital applied a specific program. The operative procedure demands that in the context of the occupational medical examination, all workers are evaluated for susceptibility to vaccine-preventable diseases (VDPs), with immunization prophylaxis offered to those determined to be susceptible. This study analyzed data from workers who attended the biological risk assessment protocol from December 2017 to October 2021 (n = 1477), who were evaluated for the immune status for measles, mumps, rubella, and varicella. Among the enrolled subjects, non-protective antibody titers were higher for measles and mumps (13%), followed by rubella (11%) and varicella (8%). Appropriate vaccinations were offered to all susceptible HCWs, and HCWs were re-tested one month after immunization. The seroconversion rate after the administration of one or more booster dose(s) was over 80%. Overall, 2.5% of the subjects refused the offered vaccine(s); the main determinant of immunization compliance was younger age (aOR = 0.86; 95%CI = 0.80-0.92). Especially during the COVID-19 pandemic, VPDs may still present a hazard in nosocomial environment. Our experience suggests that, despite hospital procedures and dedicated human assets, satisfactory VC cannot be reached without the provision of federal regulations. Nevertheless, public health policymakers have to improve the promotion of vaccine prophylaxis and education to reach higher VC.

Prevalence and management of rubella susceptibility in healthcare workers in Italy: A systematic review and meta-analysis.

Introduction: In the pre-vaccination era, all adults acquired immunity status due to natural infections during childhood and adolescence, whereas universal mass vaccination has changed the seroepidemiology of rubella among adults, showing lack of immunity in some subgroups. National and international guidelines recommend evaluating all healthcare workers (HCWs) for their immune status to rubella and possibly vaccinating those who are seronegative. We conducted a systematic review and meta-analysis to estimate the susceptibility rate to rubella among HCWs in Italy and to explore possible options for the management of those found to be susceptible. Methods: Eight studies were included in the meta-analysis, selected from scientific papers available in the MEDLINE/PubMed and Google Scholar (till page 10) databases between January 1, 2015 and November 30, 2021. The following terms were used for the search strategy: (sero* OR seroprevalence OR prevalence OR susceptibilit* OR immunit* OR immunogenict*) AND (healthcare worker* OR health personnel OR physician* OR nurse OR student*) AND (rubella OR german measles OR TORCH) AND (Italy). Results: The prevalence of rubella-susceptible HCWs was 9.0 % (95 %CI: 6.4-12.1 %). In a comparison of female vs. male serosusceptible HCWs, the RR was 0.67 (95 %CI = 0.51-0.88). Occupational medicine examinations for rubella screening with possible subsequent vaccination of seronegatives and exclusion of susceptible HCWs from high-risk settings were common management strategies. Conclusions: HCWs susceptible to rubella are an important epidemiological concern in Italy, and efforts to identify and actively offer the vaccine to this population should be increased.

Tendency in Pulmonary Aspergillosis Investigation during the COVID-19 Era: What Is Changing?

Aspergillosis is a disease caused by Aspergillus, and invasive pulmonary aspergillosis (IPA) is the most common invasive fungal infection leading to death in severely immuno-compromised patients. The literature reports Aspergillus co-infections in patients with COVID-19 (CAPA). Diagnosing CAPA clinically is complex since the symptoms are non-specific, and performing a bronchoscopy is difficult. Generally, the microbiological diagnosis of aspergillosis is based on cultural methods and on searching for the circulating antigens galactomannan and 1,3-ß-D-glucan in the bronchoalveolar lavage fluid (bGM) or serum (sGM). In this study, to verify whether the COVID-19 period has stimulated clinicians to pay greater attention to IPA in patients with respiratory tract infections, we evaluated the number of requests for GM-Ag research and the number of positive tests found during the pre-COVID-19 and COVID-19 periods. Our data show a significant upward trend in GM-Ag requests and positivity from the pre-COVID to COVID period, which is attributable in particular to the increase in IPA risk factors as a complication of COVID-19. In the COVID period, parallel to the increase in requests, the number of positive tests for GM-Ag also increased, going from 2.5% in the first period of 2020 to 12.3% in the first period of 2021.

The Geological Characteristics of the Vadose Zone Influence the Impact of Treated Wastewater on the Groundwater Quality (SCA.Re.S. Project 2019-2020).

This study evaluated whether some chemical and microbial contaminants in treated sewage effluents from two wastewater treatment plants (WWTP) reached the groundwater when they drained through a fractured karst vadose zone (WWTP-K) and a porous vadose zone (WWTP-P). Forty-five samples of sewage water (SW), treated water (TW), and monitoring well (MW), collected from WWTP-P (24) and WWTP-K (21), were analyzed for a range of microbiological and chemical properties. The E. coli and Salmonella counts were below the limits outlined in the Legislative Decree 152/06 in effluents from both types of WWTP. Enteric viruses were found in 37.5% and 12.5% of the SW and TW from WWTP-P, respectively. The percentages of Pepper mild mottle virus isolated were higher in TW (62.5% in WWTP-P, 85.7% in WWTP-K) than in SW and MW. The residual concentrations of contaminants of emerging concern (CEC) of each drug category were higher in the MW downstream of WWTP-K than of WWTP-P. Our results showed that the porous vadose zone was more effective at reducing the contaminant loads than the fractured karst one, especially the CEC, in the effluent. The legislation should include other parameters to minimize the risks from treated effluent that is discharged to soil.

Investigations on the Efficacy of Ozone as an Environmental Sanitizer in Large Supermarkets.

Awareness of the importance of the microbial contamination of air and surfaces has increased significantly during the COVID-19 pandemic. The aim of this study was to evaluate the presence of bacteria and fungi in the air and on surfaces within some critical areas of large supermarkets with and without an ozonation system. Surveys were conducted in four supermarkets belonging to the same commercial chain of an Apulian city in June 2021, of which two (A and B) were equipped with an ozonation system, and two (C and D) did not have any air-diffused remediation treatment. There was a statistically significant difference in the total bacterial count (TBC) and total fungal count (TFC) in the air between A/B and C/D supermarkets (p = 0.0042 and p = 0.0002, respectively). Regarding surfaces, a statistically significant difference in TBC emerged between A/B and C/D supermarkets (p = 0.0101). To the best of our knowledge, this is the first study evaluating the effect of ozone on commercial structures in Italy. Future investigations, supported by a multidisciplinary approach, will make it possible to deepen the knowledge on this method of sanitation, in light of any other epidemic/pandemic waves.

Microbiological and chemical characteristics of beaches along the Taranto Gulf (Ionian Sea, Southern Italy).

Coastal habitats provide important ecosystem services, such as the maintenance of ecological sustainability, water quality regulation, nutrient recycling, and sandy beaches which are important areas for recreation and tourism. The quality of seawater is generally measured by determining the concentrations of Escherichia coli and intestinal Enterococci, which might be affected by the persistent populations of these bacteria in sand. Sand might thus be a significant source of pathogen exposure to beachgoers. The quality of coastal recreational waters can also be affected by eutrophication, water discoloration, and harmful algal blooms, which pose additional human health risks. Here, we conducted a monitoring of the beaches quality along the Taranto Gulf by determining the concentrations of fecal indicator organisms, as well as other parameters that are not traditionally measured (physicochemical parameters, Pseudomonas aeruginosa, and harmful microalgae), in shallow seawater and sand sampled from three beaches. The concentrations of bacteria were determined using both standard microbiological methods and the IDEXX system. Our results demonstrate the utility of measuring a greater number of parameters in addition to those conventionally measured, as well as the importance of assessing the health risks posed by the sand matrix. Additional work is needed to develop rapid analytical techniques that could be used to monitor the microbiological parameters of solid matrices.

No-Touch Automated Disinfection System Based on Hydrogen Peroxide and Ethyl Alcohol Aerosols for Use in Healthcare Environments.

Healthcare-related infections are sustained by various bacteria and fungi. In recent years, various technologies have emerged for the sanitation of healthcare-related environments. This study evaluated the effectiveness of a no-touch disinfection system that aerosolizes 5% hydrogen peroxide and 10% ethyl alcohol. After selecting an environment, the Total Bacterial Count and the Total Fungal Count in the air and on a surface of the room were determined to evaluate the effectiveness of the aerosolization system. In addition, sterile stainless-steel plates inoculated with S. aureus, P. aeruginosa, and Aspergillus spp. isolated from hospitalized patients and reference strains were used to evaluate the effectiveness of the system. For each organism, three plates were used: A (cleaned), B (not cleaned), and C (control). The A plates were treated with non-ionic surfactant and the aerosolization system, the B plates were subjected to the aerosolization system, and the plates C were positioned outside the room that was sanitized. Following sanitization, air and surface sampling was conducted, after which, swabs were processed for bacterial and fungal enumeration. The results showed that the air sanitization system had good efficacy for both bacteria and fungi in the air and on stainless-steel plates, particularly for the A plates.

Medium-to-Long-Term Immunogenicity of BNT162b2 mRNA COVID-19 Vaccine: A Retrospective Cohort Study.

To deal with the COVID-19 pandemic, a mass vaccination campaign was started in European countries on 27 December 2020. The first vaccine available to immunize healthcare workers (HCWs) was the BNT162b2 mRNA COVID-19 vaccine. While many studies have shown a high antibody response after the second vaccine dose, antibody persistence over the medium-to-long term has yet to be evaluated. The medium-to-long-term persistence of anti-SARS-CoV-2 antibodies was determined in a sample of fully vaccinated HCWs at Bari Policlinico General Hospital, Italy. This is a observational cohort study. HCWs who completed the immunization basal cycle were screened for anti-SARS-CoV-2 IgG on days 15, 30, 60, 90, and 120 after the second vaccine dose. At each time point, >99% of the screened HCWs were seroprotected. While the geometric mean titer initially declined over time, by 60 days the titer had stabilized. Older subjects seem to lose IgG faster than younger ones. The immunogenicity conferred by the vaccine provides further evidence that it is an essential weapon in efforts to bring the COVID-19 pandemic under control. Accordingly, strict measures should be implemented, ranging from the mandatory vaccination of HCWs to strong incentives aimed at achieving vaccination of the large majority of the overall population.

Attitude for vaccination prophylaxis among pregnant women: a cross-sectional study.

Pregnant women and newborns are considered a subgroup of population at increased risk for several infectious diseases, some of which are vaccine-preventable. Anti-diphtheria-tetanus-pertussis vaccine (dTpa) and influenza vaccine are recommended for pregnant women. We carried out a study to evaluate the knowledge of new mothers toward the main vaccine-preventable diseases and to assess their compliance with recommended vaccinations. Using the Hospital Discharge Forms (SDO), the list of women who gave birth in 2018 was generated. Women were interviewed by a questionnaire administered by telephone. The study sample consisted of 145 subjects, with an average age of 35.0 ± 5.9 years (range = 18.0-47.0). 5/145 (3.4%; 95% CI = 1.1-7.9%) subjects were advised during pregnancy to have the flu shot; only 1/145 (0.7%; 95% CI = 0.5-71.6%) reported the flu vaccine during the last pregnancy. 94/145 (64.8%; 95% CI = 57.2-73.2%) respondents declared that they had carried out the TORCH panel exams before pregnancy; of these 18/94 (19.2%; 95% C = 11.8-28.6%) were susceptible for rubella. Of these subjects, for 7/18 (38.9%; 95% CI = 17.3-64.3%) rubella vaccination was offered and 5/7 (71.4%; 95% CI = 29.0-96.3%) decided to carry out the vaccination. Only 1/145 (0.7%; 95% CI = 0.0-3.8%) of the interviewed woman underwent anti-dTap vaccination. Greater efforts must be made by public health institutions to raise awareness and improve vaccination compliance in this population.

Safety profile of MenB-FHBp vaccine among adolescents: data from surveillance of Adverse Events Following Immunization in Puglia (Italy), 2018-2020.

MenB-FHBp was licensed in Europe in 2017 from the age of 10. In the "postmarketing life" of a new vaccine, surveillance of Adverse Events Following Immunization (AEFI) is crucial, to better understand the pattern of safety and the effectiveness. This paper describes the MenB-FHBp AEFIs notified in Puglia in 2018-2021, to take a picture of the safety profile of this vaccine in the real life, four years after its introduction in Italy. This is a retrospective observational study. Data were collected from the list of AEFIs notified after MenB-FHBp vaccine administration in Puglia in 2018-2020, and the number of doses of this vaccine administered in the same period. AEFIs were classified according to WHO's algorithm, and causality assessment was carried out for serious AEFIs. From January 2018 to December 2020, in Puglia, 43,061 doses of MenB-FHBp were administered and 42 MenB-FHBp AEFIs (reporting rate: 97.5 per 100,000 doses administered) were reported. Among these, 12 were classified as severe (28.6%; reporting rate 27.9 per 100,000 doses). Overall, the male/female ratio in AEFIs was 1:1. The median age of people who suffered from AEFIs was 12 years (range 11-13). For the 11 serious AEFIs for which the classification was "consistent causal association," the diagnosis was hyperpyrexia (reporting rate 13.9 per 100,000 doses), fainting (rate 4.6 per 100,000 doses), urticaria (rate 2,3 per 100,000 doses), convulsions (rate 2,3 per 100,000 doses), and vomit (rate 2,3 per 100,000 doses). No deaths or impairment were notified in studied AEFIs. The picture of MenB-FHBp vaccine supports that the risk of AEFIs is in line with previous published data and in general acceptable.

Quantitative Microbial Risk Assessment Applied to Legionella Contamination on Long-Distance Public Transport.

The quantitative microbial risk assessment (QMRA) framework is used for assessing health risk coming from pathogens in the environment. In this paper, we used QMRA to evaluate the infection risk of L. pneumophila attributable to sink usage in a toilet cabin on Italian long-distance public transportation (LDT). LDT has water distribution systems with risk points for Legionella proliferation, as well as premise plumbing for drinking water, but they are not considered for risk assessment. Monitoring data revealed that approximately 55% of water samples (217/398) were positive for L. pneumophila, and the most frequently isolated was L. pneumophila sg1 (64%, 139/217); therefore, such data were fitted to the best probability distribution function to be used as a stochastic variable in the QMRA model. Then, a sink-specific aerosolization ratio was applied to calculate the inhaled dose, also considering inhalation rate and exposure time, which were used as stochastic parameters based on literature data. At L. pneumophila sg1 concentration ≤100 CFU/L, health risk was approximately 1 infection per 1 million exposures, with an increase of up to 5 infections per 10,000 exposures when the concentrations were ≥10,000 CFU/L. Our QMRA results showed a low Legionella infection risk from faucets on LDT; however, it deserves consideration since LDT can be used by people highly susceptible for the development of a severe form of the disease, owing to their immunological status or other predisposing factors. Further investigations could also evaluate Legionella-laden aerosols from toilet flushing.

How do Vaccinators Experience the Pandemic? Lifestyle Behaviors in a Sample of Italian Public Health Workers during the COVID-19 Era.

Public health workers (PHWs) have experienced substantial workload changes because of their role in managing measures to limit the spread of COVID-19. The study's aim was to assess lifestyle changes in Italian PHWs during the pandemic. PHWs attending an annual meeting completed an anonymous questionnaire assessing their sociodemographic and behavioral characteristics and lifestyle changes during the pandemic. A total of 1000 questionnaires were completed. Most participants (63.5% women, mean age 40 ± 13.1 years) were of normal weight (61.5%), non-smokers (81.9%), had a total screen time of ≥5 h/day (83.1%), and slept at least 6 h/night (88.7%). Approximately one-third consumed sweet foods every day (30%) and did not engage in physical activity (34.6%). Current sweet food consumption, physical activity, and sleep were associated with changes in these behaviors in the last 2 years (Tau-b = 0.155; Tau-b = -0.175; Tau-b = -0.276, respectively, p < 0.001). An increase in remote working was associated with worse sleep (odds ratio (OR) 2.065, 95% confidence interval (CI) 1.482-2.877) and diet (OR 1.982, 95% CI 1.385-2.838), and increased tablet/PC use (OR 3.314, 95% CI 2.358-4.656). Health promotion measures are needed to support the adoption of healthy lifestyles in this population during the current pandemic.

An Artefactual Cluster of Mycobacterium abscessus Pneumonia among Cancer Patients Arising from Contamination.

The Mycobacteriumabscessus complex (MABC) is a group of rapidly growing, nontuberculous mycobacteria that are ubiquitous in soil, urban water pipes, swimming pools, and drinking water. Members of the MABC are considered opportunistic pathogens. The aim of this study was to investigate the origins of MABC detected in broncho-lavage (BL) samples from asymptomatic cancer patients. We turned our attention to washing and disinfection procedures for bronchoscopes; we also assessed water and disinfectant samples. Of 10 BL and 34 environmental samples tested, four BL samples (40%) and seven environmental samples (20.6%) tested positive for MABC. We hypothesized that contamination could arise from the prewashing machine and/or the water used because no patient had clinical or radiological signs consistent with MABC respiratory tract infection. Our study highlights the importance of evaluating cleaning and disinfection procedures for endoscope channels to reduce the potential spread of microorganisms and artefactual results arising from contamination.

What about Your Body Ornament? Experiences of Tattoo and Piercing among Italian Youths.

BACKGROUND: tattooing and piercing are increasingly common, especially among youths. However, several health complications may be associated with these practices if basic hygiene rules are not respected. This multicenter study was aimed at exploring tattoo and piercing experiences reported by a large sample of Italian undergraduate students through a public health perspective. METHODS: tattooed and/or pierced students attending 12 Italian universities were asked to complete a web-based questionnaire regarding their body art experience. RESULTS: out of 1472 respondents, 833 (56.6%) were tattooed and 1009 (68.5%) were pierced. The greatest proportion of tattooed students (93.9%) got her/his first tattoo in a tattoo studio, while most of the pierced were serviced in a jewelry store (48.0%). The pierced ones were less informed on health issues related to the procedure (56.0% versus 77.8% of tattooed p < 0.001), and tattooists were reportedly more attentive to hygiene rules (instrument sterilization 91.5% versus 79.1% of piercers, p < 0.001; use of disposable gloves 98.2% versus 71% of piercers, p < 0.001). CONCLUSIONS: educational interventions for both professionals and communities are needed to improve the awareness and the control of health risks related to body art throughout the Italian territory.

Could mandatory vaccination increase coverage in health-care Workers? The experience of Bari Policlinico General Hospital.

Influenza is a major issue for health-care systems all over the world and health-care workers (HCWs) are a subgroup at increased risk of exposure to respiratory pathogens, including influenza, with potential threat for both their own and their patients' health. Despite this and despite the anti-influenza vaccination being offered to all HCWs, vaccination coverage among them is still low in Europe, generally less than 30%. During 2020s influenza season, influenza vaccination begun mandatory in Puglia (Italy) and Bari Policlinico General Hospital designed an active call system in order to increase its employees' compliance to vaccination. This system was able to determine an increase of vaccination coverage from 23.1% to 77.6%.

SARS-CoV-2 and Public Transport in Italy.

Although direct contact is considered the main mode of transmission of SARS-CoV-2, environmental factors play an important role. In this study, we evaluated the presence of SARS-CoV-2 on bus and train surfaces. From the buses, we took samples from the following areas: handrails used to enter or exit the bus, stop request buttons and handles next to the seats. From the trains, the sampled surfaces were handrails used to enter or exit the train, door open/close buttons, handles next to the seats, tables and toilet handles. SARS-CoV-2 was detected on 10.7% of the tested surfaces overall, 19.3% of bus surfaces and 2% of train surfaces (p < 0.0001). On the buses, the most contaminated surfaces were the handles near the seats (12.8%), followed by door open/close buttons (12.5%) and handrails (10.5%). Of the five analyzed transport companies, bus companies were the most contaminated, in particular, companies C (40%) and B (23.3%). A greater number of positive samples were found among those taken at 10:00 a.m. and 10:55 a.m. (45% and 40%, respectively). The presence of the virus on many bus surfaces highlights how the sanitation systems on public transport currently in use are not sufficient to limit the spread of SARS-CoV-2.

Acceptance of COVID-19 Vaccination in the Elderly: A Cross-Sectional Study in Southern Italy.

In Italy, at the end of 2020, a voluntary immunization plan against COVID-19 was introduced, involving elderly among the first target categories. The aim of this study was to assess, through an online questionnaire, the acceptance of COVID-19 vaccination in a sample of older adults from southern Italy. Of a total of 1041 respondents (41.7% males, mean age 76.6 ± 6.5), 965 (92.7%) were vaccinated or willing to be vaccinated against COVID-19, although less than half of the sample was favorable to vaccinations and agreed with mandatory immunization. Acceptance of COVID-19 vaccination was found to be positively related with higher educational level (OR = 1.875, CI95% = 1.113-3.161; p = 0.018) and having social/mass media as a main source of information (OR = 2.415 CI95% = 1.358-4.296, p = 0.003). On the contrary, an inverse relationship was found between acceptance of COVID-19 vaccination and having fulfilled the questionnaire after the introduction of green pass (OR = 0.218, CI95% = 0.129-0.369; p < 0.001). Therefore, although this evidence needs to be further confirmed, it is possible to agree with previous studies reporting that compulsory measures, such as green pass implementation, must be accompanied by effective education and information strategies of the target population.