Hygiene, Volume 5, Issue 3 (September 2025) – 17 articles

Background: Dental offices and clinics utilize a variety of dental materials that are delivered in reusable containers and dispensers. However, many of these materials, including NeoPutty, BC Putty, Flowable, and Diapex, may be subject to bacterial contamination and microbial exposures from the surrounding dental office environment. Objectives: The aim of this study was to quantify and identify microbial contamination, specifically in regard to these reusable dental materials. Methods: Surfaces of new and used reusable and resealable tubes where the material dispenses and the interior surfaces of the cap were swabbed and cultured. DNA was isolated from each sample and quantitative polymerase chain reaction (qPCR) was performed to determine the presence or absence of microbial contamination, as well as the relative abundance. Results: Microbial contamination was observed among all of the “in use” samples from both the dispensing end and the interior surfaces of the cap and was strongly associated with the amount of usage. Conclusions: These data suggest that environmental contamination may be present in measurable and quantifiable amounts on reusable and resealable dental materials, which suggest the need to create protocols for sanitizing the surfaces of reusable materials to reduce the presence of microbial contamination identified in similar clinical settings. Full article

Biofilms are the source of numerous issues in the food, pharmaceutical, and production industries, making their control a major component of economic and public health. Among anti-biofilm strategies, enzyme-based products that target the biofilm matrix have proven effectiveness against multiple bacterial species. We tested the efficacy of a commercial product, Baso Bionil SL40^® (SL40; Stockmeier France, Saint-Jacques de la lande, France), against biofilms of Pseudomonas aeruginosa under various conditions of temperature, concentration, pH, and incubation time. SL40 contains two enzymes: a subtilisin protease and an α-amylase glycosidase. Our results showed that SL40 removed up to 85% of the biofilm biomass compared to tris solutions. SL40’s efficacy was strongly influenced by the presence of the enzymes and both temperature and concentration. Enzymatic activity was maintained from 20 °C to 60 °C and at pH values ranging from 5 to 9, with effective concentrations corresponding to SL40 dilutions from 3/200 to 1/200 in 50 mM tris solutions. Additionally, we observed that the P. aeruginosa biofilm biomass after pH 9 tris solution treatment was twice compared to a H₂O washing. Our results confirm the potential of enzymes against biofilms, highlight the need to define optimal application conditions, and support their integration into combined strategies for complete biofilm removal. Full article

In cemented endoprosthetics, closed prepacked mixing systems represent the most advanced generation of cementing technology. (1) Background: The purpose of the present study is to evaluate four approved prepacked systems—Palacos^® R+G pro, SmartMix™ Cemvac GHV, Optipac^® Refobacin and Cemex^® System Genta—with a focus on practical handling and intraoperative hygiene. (2) Method: The systems were evaluated according to established standard test methods for bone cements (ISO 5833), including dough time, setting time, additional mechanical tests and the level of system closure. (3) Results: The results show that all systems are safe to use and meet the general requirements, but there are relevant differences in terms of intraoperative hygiene. The Palacos R+G pro system shows significantly shorter doughing and setting times, which helps to minimize wound exposure during surgery and thus significantly reduces the overall operating time and the risk of bacterial contamination. Two of the systems cannot be classified as completely closed “pre-packaged systems.” In two cases, the system must be temporarily opened before mixing to insert the mixing element, which may result in a temporary but clinically relevant impairment of sterility and a corresponding potential risk of contamination. (4) Conclusion: From a hygienic point of view, systems that remain completely closed throughout the entire preparation process can offer advantages in terms of infection prevention. This was the case for all systems tested. Short handling times, reduced exposure of the surgical site and a shorter overall duration of the procedure could further improve intraoperative safety and reduce the risk of contamination. In terms of intraoperative hygiene, the Palacos R+G pro system achieved the best results compared to the three other systems tested due to its rapid readiness for use and comparatively short setting time (according to ISO 5833). Cemex System Genta performed worst in this respect due to its late doughing time and setting time. Full article

Hand hygiene (HH) is universally recognized as a pillar of infection prevention and control (IPC), both in hospitals and long-term care facilities such as nursing homes (NHs). An annual national survey based on the voluntary notification of purchased alcohol-based hand rubs (ABHRs) has been set up in France since 2019 to appraise the compliance to HH in NHs. The results pertaining to the Hauts-de-France (HdF) region were analyzed over the 2018–2023 period. The impact of factors such as the year, structure status, availability of an infection control link (ICL) nurse and/or an IPC team as well as constant participation in the survey was evaluated. A clear rise in the consumption of ABHRs was witnessed in 2020 followed by a progressive return to the 2018 baseline by 2023 for most NHs. This decrease was significantly less marked in NHs constantly participating in the survey. The ABHR indicator’s annual values were significantly higher in public NHs than in associative ones and in NHs with available ICL nurses/IPC teams. Finally, even though less than 50% of regional NHs reached the target of four daily ABHR uses per resident, they more frequently met it than French ones as a whole. This result underscores the need for ceaselessly reiterating the importance of HH and for pursuing training efforts in NHs with the help of ICL nurses and IPC teams. Full article

Gingivitis is a common and reversible inflammatory condition caused by dental plaque accumulation, which, if left untreated, can progress to periodontitis. Conventional oral care products like chlorhexidine (CHX) and fluoride are effective in plaque control but are often associated with adverse effects such as dental staining and mucosal irritation. This systematic review aimed to compare the efficacy and safety of natural versus conventional toothpastes and mouthwashes in managing plaque-induced gingivitis. The review followed PRISMA guidelines and was registered in PROSPERO (No. 1008296). A systematic search was conducted across PubMed, Web of Science, and Scopus for English-language clinical studies published between 2015 and 2025. Eligible studies included randomized controlled trials and clinical trials on human subjects with plaque-induced gingivitis. Exclusion criteria were studies on animals, in vitro experiments, review articles, and studies lacking control groups. Data extracted included intervention type, sample characteristics, clinical indices (PI, GI, SBI), inflammatory biomarkers, adverse events, and patient adherence. A narrative synthesis was conducted due to study heterogeneity. Fifteen studies were included. Natural products such as neem, green tea, aloe vera, and propolis demonstrated comparable effectiveness to CHX and fluoride in reducing gingival inflammation and plaque indices, with a lower incidence of side effects. In particular, natural formulations showed superior tolerability and better patient compliance, especially in long-term use. However, variability in concentration and the formulation of natural products limits their clinical standardization. In conclusion, natural oral care products appear to be effective and better-tolerated alternatives to conventional agents in managing gingivitis. Nonetheless, further long-term, standardized clinical trials are needed to confirm their efficacy and define optimal formulations. Full article

Growing concern for environmental sustainability has resulted in the implementation of sanitization methods that respect ecological principles. This research evaluates a “green” sanitizing protocol that uses CAM (Minimum Environmental Criteria)-compliant products against a traditional protocol within two ASL Roma 1 facilities. The study performed a Life Cycle Assessment (LCA) following ISO 14040, ISO 14044, and ISO 14067 standards to measure greenhouse gases emissions. Microbiological sampling was conducted according to established protocols across three different risk zones utilizing contact plates and surface swabs. The Life Cycle Assessment showed that CO₂ emissions reduced by 49.6% to 53.3% at different sites due to reduced energy use together with concentrated detergents and improved washing cycles. Microbiological testing revealed notable decreases in contamination rates across both cleaning systems yet demonstrated the “green” system achieved superior results specifically within high-risk zones. The “green” protocol matched traditional cleaning methods hygienically but delivered significant environmental advantages which positions it as a sustainable hospital cleaning solution. Full article

The efficacy of peracetic acid (0.05%, 0.5%, and 1%), sodium hypochlorite (0.2%, 0.6%, and 1%), and benzalkonium chloride (0.3%, 1.15%, and 2%) was evaluated against Staphylococcus aureus (ATCC 6538), Salmonella enterica serovar Typhimurium, (ATCC 14028), Enterococcus hirae (ATCC 8043), Pseudomonas aeruginosa (ATCC 9027), Escherichia coli (ATCC 9027), and Listeria monocytogenes (ATCC 35152) using stainless steel discs, following European Committee for Standardization (CEN) guidelines. According to CEN, a sanitizer must achieve at least a 5 Log₁₀ CFU reduction to be considered effective. Peracetic acid at 1% demonstrated the highest inactivation capacity, reducing all tested strains by more than 7 Log₁₀ CFU/mL. P. aeruginosa (ATCC 9027) showed high tolerance to sodium hypochlorite and benzalkonium chloride, with reductions below 2 Log₁₀ CFU/mL even at maximum concentrations. Both sodium hypochlorite and benzalkonium chloride, at their highest tested concentrations, effectively inactivated S. aureus, S. typhimurium, E. hirae, L. monocytogenes, and E. coli, achieving reductions greater than 7 Log₁₀ CFU/mL. Overall, sanitizers were effective only at intermediate or maximum concentrations recommended by the manufacturers, suggesting that minimum label concentrations should be avoided to ensure microbiological control. Full article

Hand hygiene is a key measure to prevent healthcare-associated infections (HAIs), yet compliance remains low in Sub-Saharan Africa (SSA), due to limited resources, insufficient training, and behavioral challenges. Simulation-based education offers a promising approach to enhance technical and non-technical skills in safe learning environments, promoting behavioral change and patient safety. This study aimed to develop and pilot a contextually adapted hand hygiene simulation-based learning scenario for nursing students in SSA. Grounded in the Medical Research Council (MRC) Framework and Design-Based Research principles, a multidisciplinary team from European and African higher education institutions (HEIs) co-created this scenario, integrating international and regional hand hygiene guidelines. Two iterative pilot cycles were conducted with expert panels, educators, and students. Data from structured observation and post-simulation questionnaires were analyzed using descriptive statistics. The results confirm the scenario’s feasibility, relevance, and educational value. The participants rated highly the clarity of learning objectives (M = 5.0, SD = 0.0) and preparatory materials (M = 4.6, SD = 0.548), reporting increased knowledge/skills and confidence and emphasizing the importance of clear roles, structured facilitation, and real-time feedback. These findings suggest that integrating simulation in health curricula could strengthen HAI prevention and control in SSA. Further research is needed to evaluate long-term outcomes and the potential for wider implementation. Full article

Introduction: Respiratory infections cause serious complications responsible for a significant number of deaths in Brazil. In addition, the causes of death can be influenced by social and economic inequalities in Brazilian regions. Objective: To analyze the epidemiological profile and the influence of demographic and socioeconomic factors on deaths from respiratory infections in the adult population between 2014 and 2023 in Brazil. Methods: This was an analytical ecological study using data from the Death Information System. Death incidences were calculated. Multinomial logistic regressions and correlation tests were used to analyze the influence of socioeconomic factors on deaths. Results: There were high incidences of deaths from unspecified pneumonia, unconfirmed tuberculosis and complicated influenza. Deaths from pneumonia and the Gini index were positively correlated, considering the variables black ethnicity (R = 0.894), age over 90 (R = 0.869) and no schooling (R = 0.818) before the pandemic. The odds ratio of death from tuberculosis and influenza in the 70–79 age group (OR = 3.97) and black ethnicity (OR = 1.24), respectively, were higher in the pandemic and post-pandemic periods compared to the previous period. Conclusions: Deaths from respiratory infections were mainly influenced by demographic variables and socioeconomic inequalities in Brazil. Full article

The term “vaccine” has been broadly and inconsistently applied to a range of products with widely divergent immunological outcomes, leading to the erosion of public trust and confusion among both medical professionals and the public. Historically, and by broad public understanding, a vaccine is expected to prevent infection, transmission, and disease through the induction of sterilizing, or true neutralizing immunity, specifically, the prevention of pathogen entry and replication in vivo. This ideal extends beyond the mere elicitation of neutralizing antibodies demonstrable in vitro. This paper proposes a three-tier classification system designed to differentiate products currently designated as “vaccines,” specifically to address the distinction between those that meet the traditional, highest-expectation definition (Tier 1), therapeutic vaccines that primarily prevent disease (Tier 2), and immunomodulatory therapeutics that primarily reduce disease severity (Tier 3). By detailing the mechanism of action of each product and emphasizing the urgent need for this refined classification, our aim is to restore public confidence in vaccination programs, improve understanding of vaccine-induced immunity among healthcare professionals, and empower informed decision-making by the public. We argue that a clearer understanding of vaccine capabilities will ultimately lead to increased vaccine uptake for those vaccines that do prevent infection, transmission, and disease. Full article

Biofilms, structured communities of microorganisms encased in an extracellular matrix, are a major cause of persistent infections, particularly when formed on medical devices. This study investigated the kinetics of biofilm formation by Escherichia coli and Pseudomonas aeruginosa, two clinically significant pathogens, on two medical-grade polymers: polyether ether ketone (PEEK) and polyamide 12 (PA12). Using a modified crystal violet staining method and spectrophotometric quantification, we evaluated biofilm development over time on polymer granules and catheter segments composed of these materials. Results revealed that PEEK surfaces supported significantly more biofilm formation than PA12, with peak accumulation observed at 24 h for both pathogens. Conversely, PA12 demonstrated reduced bacterial adhesion and lower biofilm biomass, suggesting surface characteristics less conducive to microbial colonization. Additionally, the study validated a reproducible protocol for assessing biofilm formation, providing a foundation for evaluating anti-biofilm strategies. While the assays were performed under static in vitro conditions, the findings highlight the importance of material selection and early prevention strategies in the design of infection-resistant medical devices. This work contributes to the understanding of how surface properties affect microbial adhesion and underscores the critical need for innovative surface modifications or coatings to mitigate biofilm-related healthcare risks. Full article

Parental attention and care is essential for children and adolescents who are unable to take care of their own oral health. There have been studies on the characteristics of mothers and the oral conditions of children in Korea, but there are very few previous studies that report on the oral health status of children according to the employment status of mothers. The aim of this study was to investigate the relationship between maternal employment and educational status and children’s oral health. Using data from the 10th Panel Study on Korean Children (PSKC), we analyzed the association between maternal employment and education status and the occurrence of dental caries among 1175 nine-year-old Korean children. The relationship was examined through cross-tabulation and logistic regression analysis. After adjusting for the mother’s age, parental style, parental relationship, family talk time, family meal time, leisure time, area of residence, and household income, the study found that children with working and studying mothers were 1.159 times more likely to have dental caries than children with non-working and non-studying mothers. The relationship between maternal employment and educational status and children’s oral health was confirmed. Based on the results of this study, it is expected that systematic follow-up studies will be needed to better understand the association and causal relationship between dental caries and oral disease in children according to whether mothers are employed or educated. Full article

Background: In September 2023, Storm Daniel triggered catastrophic flooding across Thessaly, in central Greece, leading to the deaths of approximately 483,476 animals and heightening concerns about zoonotic diseases, particularly Q fever caused by Coxiella burnetii. Sofades, a municipality in the Karditsa region that is severely impacted by the floods, emerged as a critical area for evaluating the risk of zoonotic disease transmission. This study aimed to determine the seroprevalence status of Coxiella burnetii Phase 1 IgA antibodies among residents in the rural area of Sofades after the Daniel floods. Methods: Serum samples were obtained from a convenient sample of residents with livestock exposure between 1 March and 31 March 2024. Enzyme-linked immunosorbent assay (ELISA) was used to detect Coxiella burnetii Phase 1 IgA antibodies. Descriptive analyses summarized demographic data, and logistic regression was employed to examine the association between gender, age, and positive ELISA results. Results: The overall seroprevalence was 16.66%. Males had a significantly higher positivity rate (28.57%) than females (6.25%). Seropositivity was more frequent among individuals aged 41–80 years, with peak prevalence observed in the 61–80 age group. Conclusions: This cross-sectional study offers a snapshot of Coxiella burnetii exposure in a high-risk rural population post-flood. The slightly higher seroprevalence in Sofades (16.66%) compared to Karditsa (16.1%) suggests limited influence of environmental factors on transmission. Despite limitations in causal inference, the findings highlight the need for enhanced surveillance and targeted public health measures. Longitudinal studies are needed to assess the long-term impact of environmental disasters on Q fever dynamics. Full article

Healthcare-associated infections (HAIs), also referred to as nosocomial or hospital-acquired infections, are a significant cause of death worldwide, with hand hygiene being the most powerful means to tackle them. The present study had a twofold aim: first, to assess the level of knowledge and the degree of compliance with hand hygiene practices based on healthcare workers’ self-reports; second, to evaluate whether behavioral economics techniques, specifically a poster combining an image and an informational message, designed according to the published literature, can increase the level of healthcare workers’ compliance with hand hygiene. Factors that potentially affect compliance were also examined. This study involved distributing a questionnaire to healthcare workers in Greek public hospitals, from which 314 completed responses were collected during the last quarter of 2023. Participants were randomly and evenly assigned to an intervention group that received a nudging poster or to a control group, which did not. Self-reported compliance with hand hygiene practices was high, based on two latent variables derived through exploratory factor analysis, although knowledge of germ transmission was moderate, as reflected in the relevant knowledge question scores. However, nudging had no statistically significant effect on hand hygiene behavior. Compliance was associated with several factors, including gender, age, work experience, profession, perceived adequacy of available hygiene-related resources, and perceived consequences of nosocomial infections on patient outcomes. Full article

In 2020, Directive (EU) 2020/2184 was published and subsequently transposed into Italian legislation via Legislative Decree 18/23 (Lgs.D. 18/23). The Directive aims to protect public health through a proactive approach based on a site-specific risk analysis along the entire water supply chain (Water Safety Plan, WSP). Between February and November 2024, a survey was conducted in Italy to assess both hospitals’ knowledge of Lgs.D. 18/23 and the application of the WSP in these facilities. A total of 300 hospitals were asked to complete an anonymous questionnaire containing 60 questions about the characteristics of the facility and the management of the water network, including the chemical–physical and microbiological monitoring of Legionella and other microbiological parameters. A total of 102 questionnaires were sent out (response rate: 34%), but only 72 were properly completed and analyzed. The results of the chemical–physical monitoring are not presented in this manuscript. Overall, 52.8% of the hospitals were built before 2000, and most are aware of Directive (EU) 2020/2184, Lgs.D.18/23 (80.6%), in particular, Article 9 on the risk assessment and management of internal water systems and the guidelines for its implementation (77.8%). All hospitals perform annual microbiological water testing, including Legionella analysis, with a detection rate of <50%. National guidelines for the implementation of WSPs are known in 75% of the hospitals, but only 38.9% have started planning to implement them, and 13.9% organize staff training on the subject. The questionnaire responses highlight the need to train hospital staff in water system risk management and WSP planning, which will be mandatory by 2029. Full article

The transmission of infectious diseases via the use of public restrooms has been previously documented. The goal of this study was to compare bacterial contamination in public vs. household restrooms and, using quantitative microbial risk assessment (QMRA), to assess the probability of infection from fomite contact with selected high-touch sites within the restrooms. Fomite surfaces in four public and four household restrooms were sampled over a period of two months. The public restrooms were in an office building occupied by 80 individuals and were considered moderate usage. The toilet seat, toilet flush handle, countertops, and floor were sampled for heterotrophic, coliform, and Escherichia coli bacteria. The highest numbers of heterotrophic bacteria and coliforms were detected on the countertops, followed by the floor. The greatest numbers of E. coli were recovered from the countertops in the household restroom, but the greatest numbers in the public restroom were recovered from the toilet flush handle. Numbers of heterotrophic bacteria and coliforms were 10 to 100 times greater in household restrooms than in public restrooms. The QMRA suggested that the greatest risk of acquiring a norovirus infection involved the touching of the countertops in household restrooms and the toilet flush handles in public restrooms. Full article

Night shift work (NSW), has been associated with adverse health outcomes in women, including increased risks of metabolic disorders, cardiovascular diseases, and reproductive dysfunctions. However, the specific effects of NSW during the climacteric period remain underexplored. This narrative review aimed to examine original studies evaluating the health impacts of NSW on women in their climacteric, with a focus on menopausal timing, hormonal balance, and related symptoms. Relevant studies published in PubMed, Embase, and Scopus were reviewed in January 2025. Of 664 articles screened, 15 met the inclusion criteria. Results indicate that NSW may lead to circadian rhythm disruption, which in turn affects hormonal regulation, the timing of menopause, and the intensity of climacteric symptoms. Prolonged exposure to night shift work appears to increase the risk of metabolic and cardiovascular conditions, and certain cancers. These findings underscore the need to consider NSW as a potential modifiable risk factor for adverse health outcomes during midlife in women. They also highlight the importance of developing targeted occupational health policies and workplace interventions to mitigate these risks and promote healthier aging and quality of life. Full article