Hygiene doi: 10.3390/hygiene6020025

Authors: Francesco Saverio Ludovichetti Roberta Gaia Parcianello Marco Rossi Valentina Brun Ernesto Comitale Davide Scettri Giulia Gargano Edoardo Stellini Sergio Mazzoleni

Background: The effectiveness of non-surgical periodontal therapy largely depends on the ability of instruments to efficiently remove plaque and calculus from root surfaces. In recent years, innovative instruments such as LM Sharp Diamond™ curettes have been introduced, featuring a coated blade that maintains sharpness without the need for resharpening. The aim of this pilot in vitro study was to compare the cleaning efficacy of LM Sharp Diamond curettes with that of conventional stainless-steel curettes, hypothesizing that the diamond-coated instruments would demonstrate superior deposit removal efficiency. Materials and Methods: Extracted teeth were selected, fixed on a support, and the root surfaces were coated with black varnish to simulate plaque and calculus deposits. Standardized photographs were taken at baseline (T0). The coated root surfaces were then instrumented using LM Sharp Diamond™ curettes (test group) or traditional Gracey curettes (control group). After treatment (T1), the images were analyzed with Image Color Summarizer software 0.82 to calculate the percentage of cleaned surface. The results were subjected to statistical analysis performed by a professional statistician. Results: The results, obtained from the treatment of nine surfaces, demonstrated that LM Sharp Diamond™ curettes achieved greater and more consistent removal efficiency, with a mean reduction of 1.27 percentage points in residual coated area (RCA) deposits compared to traditional curettes (p = 4.45 × 10−6). Conclusions: Preliminary evidence suggests that LM Sharp Diamond™ curettes may ensure more effective residue removal than traditional steel curettes. Statistically, it has been shown that LM Sharp Diamond™ curettes provide a cleaner and more uniform surface than traditional steel curettes.

Hygiene doi: 10.3390/hygiene6020024

Authors: Sebastián Loría-Palma Daniela Batista-Cárdenas Ana Paula Solís-Rivera Karol Ramírez

Background/Objectives: Electronic cigarette (e-cigarette or vape) use is increasing, especially among young adults. Since there is no data on e-cigarette use among dental students in Costa Rica, this observational study aimed to evaluate the awareness and use of vapes in students at the Faculty of Dentistry, University of Costa Rica, and identify factors that influence their use. Methods: A cross-sectional online questionnaire was administered to a sample of dental students, and their responses were assessed. A total of 244 students, with a mean age of 21.38 ± 3.29 years, were recruited between October 2024 and May 2025. Multivariate binary logistic regression was employed to analyze factors influencing ever use of e-cigarettes. Results: Of the participants, 99.17% were aware of e-cigarettes, and 60.66% had used them at least once in their lifetime, with 8.61% being current users. Sixth-year students were more likely than first-year students to believe that e-cigarettes contain carcinogenic substances (p < 0.001). Participants that were unsure if vapes are more addictive than traditional tobacco cigarettes had lower odds of ever using them (OR = 0.092, 95% CI: 0.009–0.491, p = 0.004). Fourth-year dental students had higher odds of e-cigarette use compared to first-year students (AOR = 3.050, 95% CI: 1.102–8.804, p = 0.032). The survey showed that 23.84% of the participants learned about e-cigarettes from parents, friends, or classmates, and 19.94% learned about them from social media platforms like WhatsApp, Facebook, and Instagram. Conclusions: Most of the dental students demonstrated awareness of the health impacts of vaping. While more than half of the respondents had tried e-cigarettes, current use was low. Vaping behavior was influenced by academic level and perceptions of risk and addictiveness, with social networks and peers serving as primary information sources. These findings highlight the importance of integrating targeted educational interventions into dental curricula to strengthen awareness and promote evidence-based attitudes toward vaping.

Hygiene doi: 10.3390/hygiene6020023

Authors: Livia Barenghi Alberto Barenghi

The present study proposes a narrative synthesis with an original translational approach to analyze the consequence of the global increase in waterborne microbial diseases. The focal point of this research is the relevance of these diseases for infection prevention and control (IPC) in dental settings and for public health. In order to analyze the main issues, the text focuses on studies published between January 2021 and September 2025. Over the past fifteen years, a small number of outbreaks and cases have been reported in dental settings. Nevertheless, the water utilized for dental care is frequently heavily contaminated with microbes, primarily opportunistic ones, which have the potential to cause pandemics of pseudo-infections. These include mainly Legionella, Pseudomonas, and nontuberculous Mycobacterium (NTM), antibiotic-resistant species, and other opportunistic pathogens with relative abundance exceeding 1%. This study focuses on five areas of research: (a) iatrogenic outbreaks and cases, and causes of underestimated waterborne infections; (b) the prevalence, complexity, and relevance of the dental unit water line contamination; (c) factors influencing water contamination in dental settings, (d) issues relating to products used for dental unit water line (DUWL) treatment, (e) main guidelines on water quality and European Union (EU) legislative acts and issues related to water testing. The text highlights the urgent need for greater awareness and preparedness in dental settings, as well as updated guidelines and rules to ensure the safety of patients and healthcare workers.

Hygiene doi: 10.3390/hygiene6020022

Authors: Andrea Lauková Eva Bino Jana Ščerbová Monika Pogány Simonová

Enterococci are found not only in warm-blooded animals but also as a resident population of water and an indicator of fecal pollution. The species Enterococcus haemoperoxidus and E. moraviensis are relatively new, having been detected in water. Strains with genes for biofilm formation can serve as reservoirs for gene transfer. The aim and novelty of this study were to determine whether the postbiotic substance (PS) of proteinaceous character (Durancin-like) produced by our strain Enterococcus durans ED26E/7 can inhibit in vitro the growth of biofilm-forming E. haemoperoxidus and E. moraviensis isolated from water sources. E. haemoperoxidus and E. moraviensis lacked the biofilm-forming genes ica, bap, ace, and fsrA. The bopD gene was found only in three strains of E. haemoperoxidus. The srtA gene was present in two strains of E. haemoperoxidus and E. moraviensis. Using the quantitative test, three strains showed low-grade biofilm-forming ability. They exhibited ɤ-hemolysis, and they were mostly susceptible to antibiotics. However, they were susceptible to PS Durancin-like ED26E/7. E. haemoperoxidus, the strains EHae466 and EMo494, showed the highest susceptibility to Durancin-like ED26E/7.

Hygiene doi: 10.3390/hygiene6020021

Authors: Jorge Ramsés Dorantes-Gutiérrez Jeiry Toribio-Jiménez Benjamín Maldonado-Del Moral Lorena Jacqueline Gómez-Godínez Gustavo Cuaxinque-Flores Juan Ramos-Garza José Luis Aguirre-Noyola

Foodborne diseases represent a major public health concern, particularly those associated with dairy products contaminated with Staphylococcus aureus, a pathogen capable of producing heat-stable enterotoxins. This study evaluated the potential of native lactic acid bacteria (LAB) isolated from artisanal kefir grains as natural biocontrol agents in fermented dairy foods. Kefir grains obtained from three artisanal producers were microbiologically characterized, revealing LAB as the dominant group and the absence of Enterobacteriaceae. Strains belonging mainly to the genera Lactobacillus sensu lato, Leuconostoc, and Pediococcus were isolated and exhibited differentiated metabolic profiles. Safety assessment showed no hemolytic activity and an overall susceptibility to clinically relevant antibiotics, although genus-dependent intrinsic resistance patterns were observed. Several strains displayed enzymatic activities related to carbohydrate digestion and high tolerance to simulated gastrointestinal conditions, with survival rates exceeding 90% during both gastric and intestinal phases. Neutralized cell-free supernatant (CFS) demonstrated differential inhibitory activity, with significant antagonism of S. aureus and E. coli, comparable to those of commercial reference strains. In a yogurt model system stored at 4 °C, selected Lactobacillus and Pediococcus strains induced a progressive and significant reduction in S. aureus populations, achieving complete elimination to undetectable levels in shorter times than commercial probiotic strains. Overall, these results demonstrate that native LAB from artisanal kefir grains exhibit an adequate safety and functional profile, together with strong antagonistic activity, supporting their potential application as natural protective cultures to improve the food hygiene of fermented dairy products.

Hygiene doi: 10.3390/hygiene6020020

Authors: Sebastiano Calimeri Daniela Lo Giudice Francesco Giordano Antonio Laganà Alessio Facciolà

Nurses are healthcare professionals who can play a leading role in preventing antimicrobial resistance, given their direct assistance to patients. For this reason, in-depth university training is desirable. This study was conducted to detect possible weak points in the university training about an important public health topic represented by general knowledge about antibiotics and antibiotic resistance. We carried out a survey on Knowledge, Attitudes, and Practices of students attending the Nursing Sciences course at the University of Messina, Italy, by administering an online standardised questionnaire that included general and specific questions about antibiotics and antibiotic resistance. General and specific scores were calculated. Some gaps were found about the knowledge of antibiotics (mean score: 3.6/4) and, especially, antibiotic resistance (mean score: 3.2/5). As expected, most of the incorrect answers to both antibiotic and antibiotic-resistance knowledge were given by students in the first year, but some gaps were also found among students in the last year. Given the growing role nurses can play in combating antibiotic resistance, these findings point to a possible information gap in the study course and highlight the need to enhance the current university training programmes with activities designed to increase knowledge on these important public health issues.

Hygiene doi: 10.3390/hygiene6020019

Authors: Angel McJunkin Molly Parker Kathleen Ferris Ginny Webb

Background: The emergence of alternative methods of obtaining groceries since the COVID-19 pandemic has raised new concerns regarding food safety. In this study, we sought to evaluate these concerns by evaluating how the procurement method of ground beef impacts the microbial load in the beef, as this has not been thoroughly studied. Methods: Specifically, we compared beef samples obtained from in-store shopping, grocery delivery, and meal kit delivery services to determine if these new, more convenient methods of grocery shopping impact the total microbial load or Escherichia coli present in the beef. We homogenized a total of 65 beef samples and plated dilutions on trypticase soy agar, MacConkey agar, and CHROMagar. Results: We found that in-store samples had the highest microbial load with an average of 5.06 log CFU/g, while grocery delivery samples resulted in an average of 4.76 log CFU/g and meal kit samples had an average of 4.23 log CFU/g when plated on TSA. This represents a 6.7-fold change between in-store samples and meal kit samples. These differences were not statistically significant (p = 0.1, ANOVA). When plated on MacConkey agar, in-store samples had a bacterial count at 3.0 log CFU/g, while grocery delivery samples had 2.99 log CFU/g and meal kit delivered samples had 3.05 log CFU/g. Suspected E. coli O157 colonies were detected using CHROMagar plates, as these plates function to change the coloration of positive E. coli O157 colonies to pink. Suspected E. coli O157 colonies were observed in three in-store samples, two grocery delivery samples, and one meal kit sample. After confirmatory agglutination testing, one meal kit sample was confirmed as E. coli O157. Conclusions: While trends suggest possible lower microbial contamination in delivery methods versus in-store shopping procurement, no statistical significance between methods was found. These findings indicate no significant changes in microbial loads in delivered ground beef, and the high variance suggests that all procurement methods still pose some level of risk.

Hygiene doi: 10.3390/hygiene6020018

Authors: Shinhao Yang Hsiao-Chien Huang Ying-Fang Hsu Chi-Yu Chuang

Conventional antimicrobial air filters often conflate physical interception with true biochemical inactivation, posing secondary aerosolization risks during maintenance. This study developed a lactoferricin B-functionalized polypropylene (LfCF) filter to provide a dual-action mechanism: electrostatic capture and robust contact-killing against bioaerosols. To rigorously decouple these mechanisms, a polyallylamine binder-only (PP+PAA) control was incorporated. Dynamic penetration assays at 10 cm/s revealed that the 2.0 mg LfCF achieved significantly lower viable penetration rates for Escherichia coli (41.2%) and λ phage (46.0%) compared to the PP+PAA control (75.1% and 76.3%). This substantial gap demonstrates instantaneous sublethal injury upon aerodynamic impaction, defined here as “dynamic inactivation.” Crucially, time-dependent elution assays confirmed a >2 log reduction in viable counts for both retained E. coli and λ phage on LfCFs within 60 min, definitively validating its genuine contact-killing capability. Furthermore, the amphipathic lactoferricin B peptide maintained exceptional biocidal efficacy even under high-humidity conditions (70% RH), overcoming the electrostatic shielding typical of traditional biopolymers, without increasing aerodynamic pressure drop. Finally, field validation in a dental clinic demonstrated an 83.3% reduction in airborne viable bioaerosols. As a passive, self-sterilizing engineering control, the LfCF offers a highly reliable intervention for mitigating occupational bioaerosol exposures.

Hygiene doi: 10.3390/hygiene6010017

Authors: Cesilia Charles Lutengano Mkonongo David Masanja Damian Maruba Philipo Mwita Edward Bucheye Elly Daudi Emmanuel Amsi Frank Elisha Ecka Mafwimbo Bernard Njau Nathanael Sirili Radenta Bahegwa Deogratias Banuba

Infection prevention and control remains an essential component of effective healthcare delivery and disease prevention. This study aimed to explore healthcare workers’ perspectives on factors influencing compliance with infection prevention and control practices in Katavi Regional Referral Hospital, Tanzania. With a qualitative approach, we aimed to enable a broader narrative, gain a more detailed understanding of IPC practices, and identify experiences that may be overlooked in a forced-choice questionnaire. A cross-sectional design using a phenomenological approach was employed. An interview guide was used to collect data from 19 participants (five doctors, four nurses, four laboratory practitioners, and six from administration positions; ward in-charges, quality improvement officers and administrative officers) between 24 July 2025, and 23 August 2025. Among participants, nine were the key informants, and 10 were involved in in-depth interviews. Thematic analysis revealed that the availability of IPC supplies, desire for personal and patient protection, high patient volume, awareness of IPC protocols, institutional support, supportive supervision, and HCWs’ attitudes towards IPC activities were factors influencing IPC compliance. Strengthening structured supervision, ensuring a constant supply of IPC materials, and investing in continuous IPC capacity building may be an important approach in enhancing compliance with IPC practices and reducing hospital-associated infection risk in Katavi Regional Referral Hospital and similar resource-limited healthcare settings.

Hygiene doi: 10.3390/hygiene6010016

Authors: Prem K. Sreenivasan Violet I. Haraszthy

2,6-Dichlorophenolindophenol (DCIP) is a redox dye and colorimetric reagent previously shown to enable rapid quantification of laboratory cultures of oral microorganisms. DCIP is reduced by viable microbial cells, resulting in loss of its blue color that can be measured spectrophotometrically. Previous studies demonstrated that several strains of oral bacteria and yeasts grown in culture reduce DCIP, with significant correlations observed between increasing viable plate counts and DCIP reduction. The present investigation expanded upon these studies by evaluating DCIP as a method for assessing heterogeneous mixtures of oral microorganisms collected from human subjects. Oral rinse samples were obtained from 184 adults aged 18–70 years and analyzed for DCIP reduction and viable bacterial counts. DCIP reduction was observed in all oral samples, and viable bacterial counts spanning an approximately two-log range (~100-fold difference) demonstrated a statistically significant correlation with DCIP reduction (r = 0.74; p < 0.001). Nonviable organisms did not reduce DCIP, and no DCIP reduction occurred in the absence of bacteria. These results support DCIP reduction as a practical, low-cost platform for estimating viable oral microbial burden, with the additional advantage of a visually interpretable colorimetric readout.

Hygiene doi: 10.3390/hygiene6010015

Authors: Alice Yip Wing Kiu Shek Yee Man Kiki Lee Ka Ka Lau Shuk Wai Sip Tsz Wing Lam Suet Ching Cheung Fei Lung Tang

Background: In the wake of the COVID-19 pandemic, the dietary landscapes of young adults have been profoundly reshaped. As social restrictions ease, the resurgence of dining out presents new behavioral shifts regarding health and safety. Objective: This study investigates the post-pandemic experiences of young adults in Hong Kong, focusing on the burgeoning phenomenon of eating out of home and its complex influence on eating habits and food hygiene consciousness. Methods: This qualitative study utilized a phenomenological approach to explore participants’ lived experiences. Semi-structured interviews were conducted with 12 young adults in Hong Kong to gather narratives regarding their dining practices. The data were analyzed using thematic analysis to identify recurring patterns associated with their return to public dining spaces. Results: Three core themes emerged: (i) confined palates: the remaking of the Hong Kong meal in the shadow of a pandemic; (ii) shared screen: mediating hunger from the home-as-hub; and (iii) watchful guard: the moralization of the meal amidst viral uncertainty. Conclusions: These findings dissect the critical, evolving relationship between contemporary consumption patterns and health maintenance. While the small size limits statistical generalizability, the study suggests that post-pandemic dining involves a modified reality of sustained hypervigilance. These insights offer a basis for developing sensitive and targeted public health strategies that resonate with the altered dietary realities of young adults in a post-pandemic world.

Hygiene doi: 10.3390/hygiene6010014

Authors: Luan Aparecido Alexandre Elias Marcia Cristina Nobukuni Herica Emilia Félix de Carvalho Liliane Moretti Carneiro Odinea Maria Amorim Batista Alvaro Francisco Lopes de Sousa Adriano Menis Ferreira Natália Liberato Norberto Angeloni Mara Cristina Ribeiro Furlan Marcus Felipe Calori Jorgeto Aires Garcia dos Santos Junior

To map and describe the scientific evidence on germicidal ultraviolet C (UV-C) light for hospital surface disinfection, this scoping review examined device types, reported operational parameters, microbiological and clinical outcomes, and implementation aspects. Primary studies conducted in hospital settings and evaluating UV-C or ultraviolet germicidal irradiation on environmental surfaces were searched in four databases without date restrictions. Data were synthesized descriptively in tables and narrative form following JBI and PRISMA-ScR guidance. Eleven studies (2007–2025) met the inclusion criteria. Reported microbial reductions ranged from 1 to ≥5 log10. Higher and more consistent reductions were predominantly observed under laboratory or controlled experimental conditions, whereas reductions in real-world hospital surface sampling were more variable and influenced by pathogen type, surface material, room geometry, and shadowing. Integration of UV-C with manual cleaning and multi-position irradiation cycles was associated with greater effectiveness. Reporting of key radiometric parameters (dose, exposure time, and distance) was frequently incomplete, limiting reproducibility and cross-study comparability. Clinical findings were heterogeneous: some interrupted time-series analyses suggested reductions in healthcare-associated infections, although effects were not uniform across microorganisms. Implementation reports described room-level cycle times compatible with turnover, variable staffing requirements, and limited economic evaluation. Overall, UV-C appears to be a promising adjunct to standard cleaning practices in hospital environments. However, standardized radiometric reporting, multicenter studies, and robust clinical and economic evaluations are necessary to support safe, reproducible, and sustainable large-scale implementation.

Hygiene doi: 10.3390/hygiene6010013

Authors: Rocco Loris Del Vecchio Paolo Bracciano Francesca Borghi Alessio Carminati Carolina Zellino Eleonora Pagani Giacomo Fanti Andrea Cattaneo Giovanni De Vito Andrea Spinazzè Domenico Maria Cavallo

Background: Carcinogenic, Mutagenic, and Reprotoxic (CMR) substances are among the most significant occupational health hazards in healthcare and research laboratories. Despite preventive measures and regulations, exposure assessment and risk management remain complex due to varied working practices, mixed exposures, and the lack of harmonized monitoring protocols. This systematic review investigates occupational exposure to CMR substances in laboratory and healthcare environments. Methods: Searches were conducted in PubMed, Scopus, and Web of Science up to February 2025 using tailored keyword strategies. Studies published between 2020 and 2025 reporting exposure assessment, monitoring, and/or risk management of CMR chemicals were included; non-English papers and irrelevant studies were excluded. Titles/abstracts and full texts were screened independently by two reviewers with arbitration by a third. Risk of bias was assessed by three authors who independently evaluated each study. A narrative synthesis with frequency tables was performed; no meta-analysis was conducted. Results: Of 446 screened records, 50 studies were included. Formaldehyde (25 studies) and antineoplastic drugs (18 studies) were most frequently examined. Healthcare settings—e.g., hospital pharmacies, oncology wards, and pathology laboratories—were predominant, while research laboratories were underrepresented. Inhalation was the main exposure route for formaldehyde, whereas dermal uptake and surface contamination predominated for antineoplastic drugs. Monitoring methods included air sampling, surface wipe testing, and biological assays; preventive strategies varied and were inconsistently applied. Most included studies involved environmental monitoring and did not report participant numbers, so a total number of participants cannot be aggregated; for the main outcomes, participant counts were often not available. Limitations of the evidence include marked heterogeneity across settings, matrices, analytical methods, and reporting units, which precluded meta-analysis, as well as imprecision and incomplete reporting in several studies. Conclusions: Findings reveal persistent gaps in harmonized exposure limits, monitoring standards, and long-term health surveillance, underscoring the need for comprehensive prevention strategies. This review was not registered and did not receive any external funding.

Hygiene doi: 10.3390/hygiene6010012

Authors: Kashish Rathi Nishu Devi Bharmjeet Singh Archana Ayyagari Vikram Kumar Deepti N. Chaudhari Jayesh J. Ahire

The process of ensuring the safety of the food supply is dynamic. Both the possibility of contamination and the effectiveness of safety precautions are impacted by changes in the kinds of food consumed, the geographical origins of food products, and the methods by which these foods are processed. For instance, compared to earlier generations, consumers’ general understanding of safe food preparation and handling techniques has decreased due to a higher reliance on prepackaged convenience foods. Nowadays, consumers depend increasingly on other people to make sure the food they eat is safe. Growing consumption of minimally processed foods and growing imports of fresh products from other nations have resulted from changes in consumer tastes and food processing technologies. This review aims to critically synthesize existing knowledge on microbial risks in food, focusing on their sources, mechanisms of contamination, risk evaluation methodologies, and implementation of food safety measures. Major foodborne pathogens, including Salmonella, Escherichia coli, Listeria monocytogenes, and Norovirus, are discussed alongside factors influencing their survival and transmission. Today Clostridium botulinum, Staphylococcus aureus, and Salmonella spp. remain among the major foodborne pathogens, but during the last two decades food-borne diseases such as shigellosis, listeriosis, campylobacteriosis, and diseases caused by pathogenic strains of Escherichia coli have become increasingly salient. These new concerns necessitate continued investment in research and technology development to improve the safety of the food supply. The review highlights current approaches to microbiological risk assessment, regulatory frameworks, and control strategies, while also addressing emerging challenges such as antimicrobial resistance, biofilms, and ready-to-eat foods. By integrating risk evaluation with practical implementation strategies, this review provides valuable insights for researchers, regulators, and food industry stakeholders seeking to strengthen food safety systems and reduce the burden of foodborne diseases.

Hygiene doi: 10.3390/hygiene6010011

Authors: Petra Vašaničová Kateryna Melnyk

Health and hygiene are critical components of sustainable travel and tourism development, particularly in the post-emergency phase of the COVID-19 pandemic when traveler confidence is closely tied to the resilience of the destination. This paper examines global health and hygiene conditions using data from the Travel & Tourism Development Index (TTDI) 2024, with a focus on disparities across regions and income groups. Five key indicators—physician density, basic sanitation, basic drinking water, hospital bed density, and communicable disease incidence—are analyzed to assess healthcare infrastructure, accessibility, and public health resilience. By comparing data from 2021 and 2024, the study evaluates changes during and after the peak period of the COVID-19 crisis, highlighting progress and persistent inequalities relevant to sustainable travel and tourism development. Using descriptive statistics and Spearman’s rank correlation analysis, the study also investigates the associations between key health and hygiene indicators, specifically (i) basic sanitation and basic drinking water coverage and (ii) physician density and hospital bed density, at the global, regional, and income group levels. The results reveal pronounced regional and income-related disparities. Europe and Eurasia consistently outperform other regions, with high healthcare capacity and near-universal sanitation and water access, while Sub-Saharan Africa continues to face systemic deficits in all indicators. High-income countries have well-developed healthcare systems, whereas low-income countries struggle with limited physician availability, poor sanitation coverage, and high communicable disease incidence. Associations between key indicators are also evident: countries with strong sanitation infrastructure almost always achieve high drinking water coverage, and those with higher physician density typically maintain higher hospital bed capacity. These findings highlight the uneven pace of global recovery and emphasize that health and hygiene are not only public health priorities but also fundamental drivers of tourism competitiveness.

Hygiene doi: 10.3390/hygiene6010010

Authors: Julia Kalinowski Nada Nasr Angelina Colamarino Michelle Marie Esposito

The human microbiome has emerged as a critical factor influencing surgical outcomes across a broad number of specialties. This paper explores the evolving interactions between the body’s microbiome and surgical practice, especially focusing on wound healing, post-operative recovery, and infection risk. We analyze current evidence from multiple surgical disciplines, including transplantation, reconstructive, bariatric, orthopedic, cardiothoracic, and oral surgery. Recognizing different microbial communities and their influence on surgical success opens doors to personalized medicine and targeted preventions. The studies discussed in this review provide valuable interdisciplinary insights that could ultimately uncover the microbiome’s fullest potential in optimizing future surgical protocols.

Hygiene doi: 10.3390/hygiene6010009

Authors: Cyrielle Hinson Philippe Sessou Paulin Azokpota Georges Daube Nicolas Korsak

Antimicrobial resistance (AMR) in foodborne pathogens poses a major public health risk in Benin. Table eggs and poultry sausages have been identified as potential reservoirs of resistant bacteria. Nevertheless, the lack of integrated AMR data from food and clinical sources limits the ability to assess public health risks and design evidence-based interventions. Therefore, this study aimed to determine the prevalence of Escherichia coli and Salmonella in table eggs and poultry sausages and to evaluate the antibiotic resistance profile of isolates from these foods and clinical samples alongside clinical isolates. A total of 135 table egg pools, 90 poultry sausages, and 81 clinical isolates of E. coli (56) and Salmonella (25) were collected between August and December 2023. Table eggs and poultry sausages were analysed for Escherichia coli and Salmonella using conventional methods. Antibiotic resistance (n = 99 isolates) was tested using Kirby–Bauer disc diffusion. Clinical isolates (n = 77) were included for comparison. High Escherichia coli prevalence (eggs: 59.3%, 95% CI: 50.9–67.3%; sausages: 14.4%, 95% CI: 8.6–23.2%) and lower Salmonella prevalence (eggs: 2.2%, 95% CI: 0.8–6.3%; sausages: 5.6%, 95% CI: 2.4–12.4%) were observed. According to antibiotic resistance data, predominant multidrug resistance to tetracyclines, streptomycin, and fluoroquinolones, followed by preserved sensitivity to nitrofurans and chloramphenicol, was noticed. These findings highlight the urgent need to regulate antibiotic use in Benin’s poultry and medical sectors.

Hygiene doi: 10.3390/hygiene6010008

Authors: Sandro Kresina Nataša Ivančić Jokić Vlatka Sotošek Helena Glibotić Kresina Danko Bakarčić Sara Kresina Tea Ivančić Jokić Maja Šikić Pogačar Karolina Vušković Ćosić Neda Smiljan Severinski

Background: Human papillomavirus (HPV) infection is the most common sexually transmitted infection worldwide and a major cause of several cancers. HPV vaccination is the most effective measure of preventing HPV infection, but knowledge and attitudes towards HPV vaccination are inconsistent. Methods: This study aimed to assess the knowledge, attitudes, and vaccination status related to HPV among university students in both healthcare and non-healthcare fields. A cross-sectional online survey was conducted among 769 full-time students, including 362 healthcare and 407 non-healthcare students. Validated questionnaires assessed HPV knowledge, vaccination attitudes, sociodemographic characteristics, and vaccination status. Results: Healthcare students scored significantly higher on HPV knowledge and had more positive attitudes towards vaccination compared to non-healthcare students. Vaccination rates were similar in both groups. Higher HPV knowledge was significantly correlated with reduced vaccine hesitancy and more positive vaccination attitudes. Binary logistic regression indicated that being younger, having more positive attitudes toward vaccination, and possessing greater knowledge about HPV were each associated with a significantly higher likelihood of HPV vaccination. Conclusions: In conclusion, targeted educational interventions are necessary to enhance HPV vaccination acceptance, especially among non-healthcare students. Universities represent crucial settings for promoting health literacy and increasing HPV vaccination coverage to effectively prevent HPV-related cancers among young adults.

Hygiene doi: 10.3390/hygiene6010007

Authors: Jaroslava Dekkerová Lucia Černáková

Verapamil (VER) is a calcium channel blocker used to treat cardiovascular diseases. However, some studies also suggest its antimicrobial potential. Changes in calcium abundance in yeasts can lead to decreased expression of transcription factors for genes related to morphology, resistance, and biofilm. Hyphal growth in Candida albicans is necessary for biofilm formation, especially in mixed biofilms with Staphylococcus aureus. This research studied the antibiofilm activity of VER in mixed biofilms of C. albicans SC5314 and S. aureus CCM3953. First, the minimal inhibitory concentration of VER was determined for single-species biofilms. Subsequently, a subinhibitory concentration of VER (1 mM) was tested on mixed biofilms. Biomass was reduced by 20% for C. albicans and 30% for S. aureus. The morphology of C. albicans was altered, and a decrease in S. aureus cells was also observed. qPCR was used to determine changes in HWP1 and ALS3 gene expression in biofilms formed w/wo VER. A decrease in the expression of both genes was observed. In vivo experiments with Galleria mellonella confirmed the antibiofilm activity of VER against mixed infections of C. albicans and S. aureus. These results suggest that VER regulates the morphology of C. albicans, resulting in changes in biofilm composition and the adhesion of S. aureus.

Hygiene doi: 10.3390/hygiene6010006

Authors: Gulsun Akdemir Evrendilek

Seafood processing environments represent some of the most demanding hygienic settings in the global food sector. High humidity, variable temperatures, and heavy organic residues promote the persistence of Listeria monocytogenes, Vibrio spp., and Salmonella spp., making sanitation both critical and inherently complex. This review synthesizes recent advances in hygienic design, sanitation technologies, and workforce safety as interconnected elements of a single “hygiene continuum.” Building upon Codex, FDA, and European hygiene frameworks (2020–2024), the review examines how engineering design, Sanitation Standard Operating Procedures (SSOPs) and Good Manufacturing Practices (GMPs) systems, and occupational hygiene jointly determine microbial control, sustainability, and workforce well-being. Particular focus is given to biofilm dynamics, emerging disinfection technologies, and automation through cleaning-in-place (CIP) and cleaning-out-of-place (COP) systems. Recent trends—including digital monitoring, eco-efficient cleaning, and human-centered facility design—are discussed as drivers of next-generation hygiene management. Collectively, these insights demonstrate that hygienic performance in seafood processing is not a fixed endpoint but a living system linking design, management, and human behavior toward safe, sustainable, and resilient seafood production.

Hygiene doi: 10.3390/hygiene6010005

Authors: Mark R. Limon Shaira Vita Mae G. Adviento Chariza Mae B. Basamot Jacqueline B. Reyes Karl Lorenze E. Gumsat Athena Germynne D. Amano Jessica Camille B. Ramirez Christian Jay P. Pungtilan Marie Dale R. Soriano Louwelyn B. Baclagan Shareen Kate A. Gamiao Shiella Mae G. Juan

Background: Hygiene and food-safety training is a critical public health strategy for preventing contamination and promoting safe food-handling practices in community settings. This study evaluated the long-term effectiveness of In-person, Online, and Hybrid instructional modes in enhancing hygiene and food-safety competencies among trainees in Ilocos Norte, Philippines. Methods: Using a longitudinal quasi-experimental design, performance was measured at 12, 24, and 36 months across four domains: Personal Health & Hygiene, Food Hazards, Cleaning and Sanitation, and Good Manufacturing Practices. A total of 384 students met all inclusion criteria and completed the full series of evaluations. Descriptive and inferential statistical analyses were employed. Results: Competency scores increased significantly over time in all instructional modes (p < 0.001). Hybrid learners demonstrated the highest early longitudinal gains at 12 months (mean score, M = 20.88), compared with In-person (M = 10.28) and Online (M = 10.57). At 36 months, Online learners achieved the highest performance (M = 19.50), indicating stronger long-term retention. Effect size analysis using eta squared (η2) showed large effects for Cleaning and Sanitation (η2 = 0.196), Good Manufacturing Practices (η2 = 0.115), and overall performance (η2 = 0.138). Standardized Mean Change (SMC) indicated substantial improvement across modes, with Hybrid showing the greatest early change (SMC = 41.76 at 12 months) and Online exhibiting the strongest long-term improvement (SMC = 38.80 at 36 months). Training Efficiency Index (TEI) identified In-person instruction as most efficient (TEI = 30.55), followed by Online (29.49) and Hybrid (19.56). Linear Mixed-Effects Regression confirmed significant main effects of Time (β = 4.82, p < 0.001) and Mode (β = 3.97, p < 0.001), as well as a significant Time × Mode interaction (β = −1.42, p < 0.01). Conclusions: The findings indicate that Hybrid instruction supports rapid early competency gains, while Online instruction yields superior long-term mastery of hygiene and food-safety competencies. These results provide evidence-based guidance for optimizing hygiene training programs in community and public health contexts.

Hygiene doi: 10.3390/hygiene6010004

Authors: Eirini Thanasi Maria Antoniadou Petros Galanis Vasiliki Kapaki

Background: Despite its crucial role in overall health, oral health is frequently overlooked within healthcare systems, partly due to the misconception that oral diseases are neither life-threatening nor directly disabling. This perception has led to an underestimation of the psychological, social, and economic burden associated with oral diseases. Τhe present study aimed to assess oral health status and oral health-related quality of life among dental patients attending a public Special Care Center in Greece. Methods: A cross-sectional study was conducted among 400 dental patients aged 18 years and older who visited a public Special Care Center for a routine check-up or a dental problem between September and October 2024. Data was collected through personal interviews and clinical examinations after informed consent was obtained. Oral health-related quality of life was evaluated using the Oral Health Impact Profile-14 (OHIP-14) and the Oral Impacts on Daily Performance (OIDP) questionnaires. Categorical variables were presented as absolute and relative frequencies, while quantitative variables were summarized as mean, standard deviation, median, minimum, and maximum. Normality was assessed using the Kolmogorov–Smirnov test. Bivariate analyses and multivariate linear regression models were performed, with statistical significance set at p < 0.05. Statistical analyses were conducted using IBM SPSS 23.0. Results: The majority of participants were female (56.3%) with a mean age of 50.4 years (SD = 14.9). Overall oral health-related quality of life was moderate (OHIP-14: Mean = 21.0, SD = 14.8; OIDP: Mean = 14.0, SD = 12.8). Patients who attended the center due to a dental problem reported significantly poorer oral health outcomes than those attending routine check-ups (p < 0.001). Poorer self-rated oral health, having ≥12 missing teeth, prosthetic restoration, and foreign nationality were significantly associated with worse oral health-related quality of life. Conclusions: Dental patients attending the Special Care Center demonstrated moderate oral health status, which was associated with psychological distress, physical disability, and social limitations. These findings underline the need for targeted public oral health interventions, especially for vulnerable population groups.

Hygiene doi: 10.3390/hygiene6010003

Authors: Tasmiya Rangila Andiswa Zondo Andiswa Mtshali Najiha Ismail Suleman Tar Uzair Shabbir Dada Etando Ayukafangha Akebe Luther King Abia

Background/Objectives: Although children’s playgrounds foster physical, cognitive and emotional health, sandpits can harbour antibiotic-resistant bacteria, representing a health concern for kids. Therefore, this point prevalence study investigated the presence and antimicrobial resistance of Escherichia coli in sandpits at four schools in Durban to ascertain the potential risk to schoolchildren and inform school authorities of the need to prevent such occurrences. Methods: Twenty samples were collected from schools on a single day. E. coli was isolated using colilert-18® and confirmed using PCR. Antibiotic susceptibility testing was performed against 19 antibiotics using the disc diffusion method and Clinical and Laboratory Standards Institute (CLSI) guidelines. Results: E. coli was detected in 2/4 schools (50%), yielding 100 pure isolates. Of these, 71% (31 Site B and 40 Site C isolates) were resistant to at least one of the antibiotics tested, displaying 36 antibiograms. The highest resistance was to CFX (n = 40), and the lowest was to AMK and MEM (n = 1). All isolates were susceptible to CIP, CHL, GEN and TZP. At Site B, the highest resistance was against CFX (n = 16) and the lowest against AMK, CTX and NAL (n = 1). The highest resistance at Site C was against TET (n = 26), and the lowest against ATH and AUG (n = 1). Twenty isolates (20%) were multidrug-resistant, displaying resistance to at least one antibiotic from 3 classes. Conclusions: These results show that children with poor hygiene practices could get sick from playing in sandpits. Schools must change their sand regularly and ensure that sandpits are constantly exposed to the sun.

Hygiene doi: 10.3390/hygiene6010002

Authors: Ana Rita Barata Beatriz Ferreira Patrícia Oliveira Hugo Guedes Maria José Saavedra Gonçalo Almeida

Background: Domestic refrigeration and egg handling are key factors in ensuring household food safety. Inadequate temperature control and poor hygiene in refrigerators can promote the survival and growth of foodborne pathogens. This study aimed to (i) characterize refrigerator temperature profiles and surface microbial contamination and (ii) screen eggs and egg-storage areas for the presence of Salmonella spp. and Campylobacter spp. Methods: Fifty domestic refrigerators were monitored twice in 2024 and 2025 in Porto, Portugal. The temperatures were continuously logged on the lowest shelf, which was swabbed for microbiological analysis. Surface hygiene was evaluated using total viable counts (TVC), Enterobacteriaceae, and Escherichia coli enumerated following ISO methods. Detection of pathogens Listeria monocytogenes, Salmonella spp., and Campylobacter spp. was performed using real-time PCR. Eggs (n = 92 in 2024; n = 88 in 2025), and domestic egg storage areas (total n = 76) were screened for Salmonella and Campylobacter. Results: The mean refrigerator temperatures were 6.0 ± 0.5 °C in 2024 and 6.1 ± 0.5 °C in 2025; 44% and 50% of the units, respectively, exceeded the recommended 6 °C threshold. In 2025, 31 (62%) and 33 (66%) refrigerators showed higher TVC and Enterobacteriaceae counts compared to 2024, whereas E. coli was only detected sporadically. L. monocytogenes, Salmonella spp., or Campylobacter spp. were not recovered from the refrigerator surfaces. Likewise, Salmonella and Campylobacter were not detected in any of the eggs or egg-storage sites. Indicator microorganism’s counts were not associated with the mean temperature. Conclusions: The absence of correlation between ΔT and Δ microbial counts suggests that behaviour-driven hygiene factors, rather than the relatively small year-to-year temperature differences observed, are more influential in determining household bioburden. Maintaining refrigerator temperatures ≤ 6 °C together with simple hygiene practices remains essential for reducing household food safety risks.

Hygiene doi: 10.3390/hygiene6010001

Authors: Soraia Salman Josy Goldoni Lazarini Daniel Saraiva Lopes Tatiane Tiemi Macedo Diego Romario-Silva Maria Ligia Rodrigues Macedo Pedro Luiz Rosalen Rosemary Matias Severino Matias de Alencar Janaina de Cássia Orlandi Sardi

Background: Tracheostomy procedures are associated with increased risk of nosocomial infections due to microbial colonization and biofilm formation on tube surfaces. These biofilms contribute to persistent infections and hinder clinical recovery. Plant-derived products have gained interest as alternative strategies for preventing device-associated infections. Methods: This study evaluated the phytochemical composition and the antimicrobial, anti-adherent, antibiofilm, anti-inflammatory, antioxidant, and toxicity properties of Tapirira obtusa bark extract (TOBE). Antimicrobial activity was determined by minimum inhibitory concentrations (MICs). Biofilm formation and microbial viability were assessed in mono- and mixed-species biofilms. Anti-inflammatory effects were evaluated by NF-κB inhibition and TNF-α quantification. Antioxidant activity was measured using the DPPH assay. Phytochemical analysis identified major bioactive groups, and toxicity was tested in the Galleria mellonella model. Results: TOBE exhibited notable antimicrobial activity, with MIC values between 3.9 and 31.25 µg/mL. At 78 µg/mL, the extract significantly reduced biofilm biomass and microbial viability (p < 0.05). TOBE also downregulated NF-κB activation and decreased TNF-α levels. Antioxidant assays confirmed radical-scavenging capacity. Phytochemical screening revealed phenolics, flavonoids, and tannins, and toxicity results indicated a safe profile. Conclusion: TOBE effectively inhibits microbial growth and biofilm development on orotracheal tube surfaces while exhibiting anti-inflammatory and antioxidant properties without detectable toxicity. These findings support its potential as a plant-based adjunct for preventing tracheostomy-related infections and improving patient outcomes.

Hygiene doi: 10.3390/hygiene5040060

Authors: Antonia Mataragka Rafaila Anthi Zoi-Eleni Christodouli Olga Malisova Nikolaos D. Andritsos

Ensuring optimal food hygiene is essential for food safety and preventing foodborne illness, although the importance of food hygiene is often overlooked in the household kitchen setting. Adequate, good hygiene practices in the domestic environment are equally important as their implementation in any other food preparation environment, like in the food industry. The current review encompasses research data on the prevalence and isolation of major foodborne pathogenic bacteria (Campylobacter, Salmonella, Listeria monocytogenes, Staphylococcus aureus, Escherichia coli pathotypes, and Clostridium perfringens) from household kitchen equipment, as well as food cleaning utensils used in the kitchen, such as sponges, brushes, dishcloths, and hand towels. The most common bacterial pathogen present in the domestic environment is S. aureus. The latter can be transmitted orally, either via direct hand contact with contaminated kitchen surfaces and/or cleaning utensils, or indirectly through the consumption of contaminated food due to cross-contamination during food preparation (e.g., portioning prepared meat on the same cutting board surface and with the same knife previously used to cut fresh leafy vegetables). Moreover, research findings on the hygiene of food cleaning utensils demonstrate that (i) sponges have the highest microbial load compared to all other cleaning utensils, (ii) brushes are less contaminated and more hygienic than sponges, thus safer for cleaning cutlery and kitchen utensils, and (iii) kitchen dishcloths and hand towels positively contribute to cross-contamination since they are frequently used for multiple purposes at the same time (e.g., drying hands and wiping/removing excess moisture from dishes). Finally, the present review clearly addresses the emerging issue of antimicrobial resistance (AMR) in bacterial pathogens and the role of the domestic kitchen environment in AMR dissemination. These issues add complexity to foodborne risk management, linking household practices to broader AMR stewardship initiatives.

Hygiene doi: 10.3390/hygiene5040059

Authors: Gabriele Poletti Alessia Mariani Stefano Brovarone Alessandro Libretti Livio Leo Bianca Masturzo Alessandro Messina

The central venous catheter (CVC) is essential in the management of pediatric patients, allowing the administration of medications, parenteral nutrition, and other treatments. However, its use carries a high risk of central-line-associated bloodstream infections (CLABSI) and catheter-related bloodstream infections (CRBSI). Advanced chlorhexidine-impregnated dressings have been developed to reduce bacterial colonization, but their effectiveness in the pediatric population remains uncertain. The aim of this review is to evaluate the effectiveness of chlorhexidine-impregnated dressings compared to standard dressings in reducing CLABSI, CRBSI, and CVC colonization in pediatric patients. Randomized clinical trials published between 2005 and 2021 in PubMed, CINAHL, and Embase, in Italian or English, were selected. Methodological quality was assessed using the Joanna Briggs Institute (JBI) checklist. Relevant data were extracted and summarized in tables. Four studies, including a total of 733 pediatric patients (367 intervention, 366 control), were included. None reported a statistically significant reduction in CLABSI/CRBSI with the use of chlorhexidine-impregnated dressings. However, two studies showed a significant reduction in catheter colonization in the intervention group. Current evidence does not support a superior effect of chlorhexidine-impregnated dressings in preventing CLABSI/CRBSI in pediatric patients, although they may reduce catheter colonization. Randomized trials with larger samples and specific methodologies are needed to clarify the true clinical impact.

Hygiene doi: 10.3390/hygiene5040058

Authors: Anuli Njoku Mousa Al-Hassan Sharaban Tohura Kayla Garcia

Period poverty refers to the lack of access to or affordability of menstrual hygiene supplies such as sanitary products and the inaccessibility of washing facilities, waste disposal and educational materials. Period poverty can significantly affect menstruating individuals’ physical, mental, and reproductive health and emotional wellbeing; negatively impact educational outcomes; cause financial strain; result in absenteeism from work and school; create barriers to healthcare access; and perpetuate poor health outcomes for generations. Barriers to menstrual equity include lack of access to period support, cost, poor sanitary facilities, lack of education, social and cultural stigma, and legal restrictions. Therefore, it is crucial to actively advocate for initiatives to increase access to menstrual hygiene products, raise public awareness, and educate individuals on safe menstrual practices. Approximately 500 million girls and women worldwide and an estimated 16.9 million people in the United States experience period poverty, with the issue being particularly common among marginalized groups such as Black or Hispanic menstruating individuals and those who are homeless, living in poverty, of low income, or attending college. This article investigates the physical, psychological, educational and social impacts of inequitable access to menstrual products, menstrual education, and sanitation facilities among menstruating individuals who are Black, Hispanic or of low income within the United States. We examine the threat this poses to health equity and propose recommendations to address this pervasive issue.

Hygiene doi: 10.3390/hygiene5040057

Authors: Shareen Kate A. Gamiao Marie Dale R. Soriano Realyn Q. Salvador Ian Mark A. Catacutan Hanna Jeoselle A. Padua Shaira Vita Mae G. Adviento Tricia Mae A. Sagucio Ann Kirsten C. Malapit Mark R. Limon

Background: In the Philippine basic education system, particularly in Technology and Livelihood Education (TLE) Cookery classes, teachers are at the forefront of promoting proper food hygiene. However, systemic challenges such as the absence of standardized policies, outdated training, and lack of resources hinder their effectiveness. To address this gap, this study aimed to assess the food hygiene knowledge and practices of cookery teachers and provides localized evidence to address the lack of division-level hygiene policies. Methods: A descriptive research design supported by qualitative interviews was employed. In total, 69 junior and senior high school cookery teachers from three school divisions in Ilocos Norte participated. A researcher-made survey questionnaire and an interview guide were used to gather data, which were analyzed using descriptive and inferential statistics. Participant testimonies were integrated to enrich the quantitative findings. Results showed high levels of food hygiene knowledge (x¯= 3.48; Highly Competent) and practices (x¯= 3.80; Highly Competent). Despite these strengths, notable gaps were identified in technical areas such as temperature control, cold storage, and specific aspects of personal hygiene. Conclusions: Interviews highlighted the need for a formal institutional hygiene policy to support teachers’ implementation of safe food-handling practices. All computed p-values were below 0.01, indicating significant correlations between demographic variables and both knowledge and practices. The correlation values ranged from r = 0.039 to r = 0.342, suggesting weak to moderate positive relationships and indicating that hygiene behaviors are influenced by multiple factors rather than demographics alone. Based on the findings, the study recommends institutionalizing the proposed policy brief, providing adequate resources, and implementing continuous professional development for Cookery teachers. The study’s scope is limited to Northern Philippines.

Hygiene doi: 10.3390/hygiene5040056

Authors: Hye-Min Ku Jong-Woo Kim Mi-Kyoung Jun

Background: The use of computer-aided design and manufacturing (CAD/CAM) technology is rapidly expanding in modern dentistry. However, differences may exist in educational experiences and utilization competencies between dental hygiene and dental technology students. Objectives: This study aimed to assess CAD/CAM educational exposure and competencies by major and grade level. Methods: A cross-sectional online survey was conducted with 464 undergraduate students in Korea. The questionnaire covered general characteristics, CAD/CAM education experience, and competencies in scanning, CAD design, CAM operation, and post-processing. Data were analyzed using descriptive statistics, independent-samples t-tests, and correlation analysis. Results: A total of 464 valid responses were analyzed. Dental technology students reported significantly higher experience with CAD/CAM education compared with dental hygiene students (p < 0.001). Scanning, CAD design, CAM operation, and post-processing competency were all significantly higher among dental technology students and senior (3–4 year) students compared with dental hygiene and junior (1–2 year) students (p < 0.001). CAD/CAM competencies varied significantly by major and grade level. Conclusions: These findings emphasize the need to strengthen CAD/CAM training in dentistry curricula and to promote interdisciplinary digital dentistry education to prepare practice-ready graduates for a digitized clinical environment.

Hygiene doi: 10.3390/hygiene5040055

Authors: Mapula Luckyjane Molewa Lee Heine Tobias George Barnard Nisha Naicker

Background: Environmental surfaces are key reservoirs for pathogen transmission, with the survival of bacteria on fomites influenced by factors such as temperature, humidity, and microbial interactions. This study aimed to determine microbial surface contamination and to determine the antimicrobial resistance profile of bacteria isolated from the indoor surface where the presence of cockroaches was observed in households of the Greater Letaba Municipality (GLM), South Africa. Methods: Swab samples were collected from kitchen countertops and food storage areas with visible cockroach activity. Bacteria were isolated and identified using standard microbiological methods, and antimicrobial susceptibility testing (AST) was conducted with the Vitek® Automated 2 system. Results: Of the 120 samples collected, 82 (68%) showed bacterial growth, resulting in 190 isolates. The majority of isolates (93%) were Gram-negative, comprising Klebsiella, Pseudomonas, Enterobacter, Escherichia, Serratia, Stenotrophomonas, Pantoea, Raoultella, and Salmonella species, with 98% demonstrating multidrug resistance (MDR) to multiple antibiotics. Resistance was particularly high against gentamicin (94%), fluoroquinolones (88%) and amikacin (77%). Among Gram-positive isolates, all belonged to the Enterococcus species, with 22% being resistant to one or two of the tested antimicrobial agents and 78% exhibiting MDR. Conclusions: The study revealed a high prevalence of antibiotic resistance in both Gram-positive and Gram-negative bacteria isolated from household surfaces. The spread of antibiotic-resistant pathogens via environmental surfaces presents a significant risk to human health, safety, and well-being.

Hygiene doi: 10.3390/hygiene5040054

Authors: Manuela Kirchner Cora Delling Arwid Daugschies

Background: The cysts of the protozoan parasite Giardia duodenalis, which targets a broad spectrum of hosts including humans, can withstand environmental conditions for months, making effective disinfectant measures crucial for minimizing the infection burden. Previous investigations concerning disinfection efficacy were based on cysts from fecal/water samples or animal models, which are either unfit for standardized procedures or related to ethical concerns. Methods: To perform standardized in vitro disinfectant testing, four different encystation protocols were compared firstly. The protocol with the highest efficacy in our hands (1.7 × 105 cysts per tube) was used for the production of cysts to establish a disinfectant assay. Therefore, it was used for the production of cysts to establish a dis-infectant efficacy assay. After incubation with a commercial disinfectant (ViPiBaX Giardien Ex®) or 30% hydrogen peroxide solution (H2O2) at 10 °C and room temperature, parasite cyst viability was evaluated by the yield of trophozoites obtained by the applied excystation protocol. Results: Only untreated Giardia cysts, which were used as a negative con-trol, released trophozoites. The protocol established for the evaluation of cyst viability delivered reproducible results and appeared suitable for testing the inactivation of cysts by chemical disinfection. Conclusions: Under the given conditions, the disinfectant ViPiBaX Giardien Ex® and H2O2 inactivated Giardia cysts.

Hygiene doi: 10.3390/hygiene5040053

Authors: Lissa Aoki Juliana de Carvalho Rodrigues Ingrid Bertollini Lamy Glaucia Luciano da Veiga Beatriz da Costa Aguiar Alves Edimar Cristiano Pereira Luiz Vinicius de Alcantara Sousa Fernando Luiz Affonso Fonseca

Introduction: The COVID-19 pandemic disrupted social interactions, family dynamics, and economic stability, disproportionately affecting vulnerable populations. Tuberculosis and leprosy perpetuate poverty and, once manifested, hinder socioeconomic development due to their high potential for disability. Methodology: This study analyzed the impact of the COVID-19 pandemic using DATASUS health data and assessed the influence of socioeconomic interventions (SAGICAD data) on tuberculosis and leprosy case notifications in Brazil. A correlation analysis was performed between regional diagnoses and variables such as Bolsa Família (a national social welfare program), BCG vaccination coverage, and COVID-19 immunization rates, applying Pearson’s correlation test. Results: No significant correlations were found between COVID-19 vaccination rates and tuberculosis/leprosy diagnoses. However, a strong negative correlation (p < 0.05) was observed between BCG (Bacillus Calmette–Guérin) vaccination coverage and leprosy incidence in the Northern region. The findings also suggest that social assistance programs such as Bolsa Família play a pivotal role in preventing infectious diseases in vulnerable areas. Conclusions: Understanding the complex interplay between socioeconomic determinants and public health outcomes is essential for guiding future research and informing health policies, including potential revisions to social programs and vaccination protocols.

Hygiene doi: 10.3390/hygiene5040052

Authors: Lindita Aliaga Armend Aliaga Katarina Smilkov Zorica Naumovska Marija Darkovska Serafimovska Darinka Gjorgieva Ackova

Medication non-adherence remains a major challenge in healthcare systems worldwide, with significant implications for patient outcomes and healthcare costs. This study explored the perceptions and practices of Key Opinion Creators (KOCs) in Kosovo regarding medication adherence. Using a qualitative approach, 15 participants representing hospitals, pharmacies, academia, pharmaceutical companies, insurance, and professional associations were surveyed through open-ended questionnaires. Findings reveal the absence of clear national policies or guidelines dedicated to adherence, limited monitoring systems, and insufficient institutional support. While KOCs recognize healthcare professionals’ (HPs) role in patient education, counseling, and monitoring, they also stressed that their engagement is often fragmented and lacks standardized reporting mechanisms. Technology and innovation were identified as promising tools to improve adherence, but their application is limited by inadequate infrastructure and resources. Key barriers include a lack of guidelines, financial and human resource constraints, low patient education, and disparities across demographic groups. The study highlights the urgent need for coordinated policies, stronger inter-professional collaboration, targeted interventions, and integration of digital health solutions to strengthen medication adherence practices in Kosovo.

Hygiene doi: 10.3390/hygiene5040051

Authors: Camila Carlino-Costa Marco Antonio de Andrade Belo

Sustainable aquaculture is increasingly vital to meet global protein demands while ensuring fish product safety and environmental stewardship from a One Health perspective. This review addresses fish hygiene as a comprehensive, multi-stage challenge encompassing water quality management, pathogen control, antimicrobial stewardship, feeding practices, humane slaughter, post-harvest handling, and monitoring systems. We examined current practices and technologies that promote hygienic standards and reduce contamination risks across production cycles. The integration of biosecurity measures and alternative health-promoting agents contributes to disease prevention and reduces reliance on antimicrobials. Responsible drug administration aligned with regulatory frameworks minimizes residues and antimicrobial resistance. Feeding strategies incorporating sustainable and safe ingredients further support fish health and product quality. Critical control points during slaughter and post-harvest processing ensure microbial safety and prolong shelf life. Advanced monitoring and traceability systems enable real-time oversight and enhance food safety assurance. Finally, certification programs and robust regulatory policies are essential to standardize practices and facilitate access to international markets. Collectively, these strategies foster sustainable aquaculture that safeguards public health, maintains ecological integrity, and supports economic viability. This holistic approach positions fish hygiene not as a final quality check, but as an integral, continuously managed component of responsible aquaculture production.

Hygiene doi: 10.3390/hygiene5040050

Authors: Dachel Martínez Asanza Anuli Njoku Snehal Baviskar Marian A. Evans Kamel Mouloudj

Background: There is a need to develop comprehensive guidelines to encourage the promotion of oral hygiene care among older adults and to assist caregivers in this endeavor, taking into consideration the specific challenges that arise from aging, comorbidities and caregiving. Methods: This review was conducted by searching across relevant literature from meta-databases including Academic Google, PubMed, Scielo and Scopus for studies published from 2020 to 2024. PRISMA guidelines were followed. We included articles that described oral hygiene methods, caregiver education and mechanization status of older adults. Common themes, best practices, and gaps in current guidelines were tracked using extracted and analyzed data. Results: The review revealed multiple factors affecting the oral hygiene of older adults, with themes relating to physical impairment, cognitive dysfunction, and caregiver involvement. Highlighted between the approaches are individualized therapy for oral hygiene, caregiver education, and the use of technology to improve adherence to oral hygiene. Barriers like dental care access, underlying medical conditions complicating dental treatments, and cost considerations were identified. Conclusions: The findings emphasize the necessity of clear recommendations that can help caregivers and advance dental care for older adults.

Hygiene doi: 10.3390/hygiene5040049

Authors: Nada Hussien Ielewi Faiza M. Abdul-Ameer

Background: Denture stomatitis (DS) represents an oral fungal infection induced by Candida albicans, impacting approximately 70% of the individuals who use removable acrylic dentures. Researchers suggest that the high level of the Candida species, particularly Candida albicans (C. albicans), is the predominant etiological factor of DS. Consequently, the development of a soft liner with antifungal activity might significantly enhance its therapeutic applicability. This in vitro study evaluates the impact of phosphate bioactive glass reinforced heat-cured acrylic-based soft liner on the candidal activity in this material. Method: Specimens (10 mm × 2 mm disc-like) were required for the selected test; PBG-Sr nano-powder was synthesized and added to the soft liner at percentages of 1 wt.%, 3 wt.%, 5 wt.%, and 7 wt.%. The candidal adherence test was investigated, and characterization was performed by X-ray diffraction analysis, field emission scanning electron microscopy, energy dispersive X-ray spectroscopy mapping, and particle size analysis. The resulting data were analyzed with one-way ANOVA followed by Dunnett’s test. Results: Candidal adherence in the 1 wt.%, 3 wt.%, 5 wt.%, and 7 wt.% PBG-Sr subgroups had decreased values in comparison to the control (0 wt.%), with the 7 wt.% subgroup demonstrating the lowest count of C. albicans (0.027), close to the nystatin group. Conclusions: PBG-Sr can diminish C. albicans adhesion in soft lining materials, and a soft liner containing PBG-Sr (7 wt.%) showed the most effective activity against C. albicans in the soft liner. Soft liners infused with bioactive glass may have the potential to assist those struggling with denture stomatitis, providing patients with enhanced therapeutic qualities.

Hygiene doi: 10.3390/hygiene5040048

Authors: Anna Rita Corvino Elpidio Maria Garzillo Daniele Guida Simona Signoriello Giuseppe Porciello Antonio Arnese Antonella Palucci Monica Lamberti

Healthcare workers, particularly those in training, face significant exposure to biological risks, including needlestick and sharps injuries, which remain a critical occupational hazard. This study examines the incidence and nature of such injuries among young healthcare trainees at the University of Campania “L.Vanvitelli,” with the aim of providing useful information for designing an effective preventive program. From 2014 to 2023, a study population of 14,908 health students was subject to health surveillance, of which 141 workers suffered an occupational injury. Needlestick injuries were most common (67.4%). Surgical residents were the most affected subgroup (24.8%). Injuries occurred more frequently after the first year of training (74.5%). Multivariate logistic regression showed significantly higher odds of injury for nursing students (OR = 8.673; 95% CI: 3.196–35.634) and residents (OR = 50.726; 95% CI: 17.789–214.279) compared to medical students. The findings underscore the urgent need for enhanced risk management strategies, targeted education programs, and stricter adherence to safety regulations to protect this vulnerable population.

Hygiene doi: 10.3390/hygiene5040047

Authors: Myoung-Hee Kim Ho-Jin Jeong Young Sun Hwang

Background: With the increasing popularity and diversification of cosmetic products, an expanding number of retail stores are providing in-store testers to allow consumers to try products before purchasing. However, growing concerns have been raised about microbial contamination and the associated risk of infection due to the shared use of these testers. This study aimed to investigate consumer awareness and perceived susceptibility to microbial infection associated with the use of lip cosmetic testers, and to identify major pathogenic bacteria, thereby offering evidence to support the need for improved hygiene management practices. Methods: The survey examined the use of lip cosmetic testers and related side effects, as well as perceptions of product hygiene and microbial contamination. Awareness of microbial infection and consumer sensitivity to product safety were measured using a five-point Likert scale. Microorganisms were detected through PCR analysis of genomic DNA. Results: Among 134 respondents, 95% had previously used lip cosmetic testers, and 90% recognized the potential for microbial contamination. Sensitivity to product safety was not significantly associated with sociodemographic factors. However, susceptibility to microbial infection from tester use was significantly higher among participants of older age and with higher education levels. PCR analysis of lip cosmetic testers revealed frequent detection of Staphylococcus epidermidis and Pseudomonas aeruginosa. Other detected bacteria included Streptococcus oralis, Streptococcus salivarius, Streptococcus pneumoniae, and Escherichia coli, all of which are known to cause oral and respiratory infections. Conclusions: The analysis revealed that the majority of individuals who have used lip cosmetic testers expressed a high level of concern regarding product contamination and the potential for infection. Multiple pathogenic bacteria linked to oral and respiratory diseases were identified in the opened tester products. These findings underscore the necessity of establishing regulatory guidelines for the management of lip cosmetic testers and highlight the importance of enhancing consumer awareness regarding hygiene practices.

Hygiene doi: 10.3390/hygiene5040046

Authors: Darío de Jesús Guillén-Morales Isabel Cruz-Cortés Taurino Amilcar Sosa-Velazco Alba Soledad Aquino-Domínguez

Breastfeeding is a complex biological system and a bidirectional physiological dialogue in which the infant may contribute to maternal breast health. This review synthesizes current evidence, clearly separating established findings from emerging hypotheses, to examine the possible infant-driven mechanisms that influence hormonal and immune homeostasis in the mammary gland. We evaluate how neonatal suckling coordinates interconnected hormonal reflexes and immune activity, and we explore the hypothesis that the retrograde flow of infant saliva to the breast tissue could activate maternal enzymatic defenses, particularly the xanthine oxidase and lactoperoxidase systems. We also consider the activation of antimicrobial peptides through direct contact at the nipple and areola, including cathelicidin and defensins, as well as the potential roles of fetal microchimerism and microbial transfer from the infant’s mouth in strengthening breast resilience. Although much of the evidence remains indirect and based on in vitro and animal models, the convergence of data supports a reformulated conceptual model that presents the infant as an active physiological partner rather than a passive recipient of milk. Recognizing this shift has important clinical implications for the prevention of inflammatory conditions such as mastitis, the improvement of breastfeeding support strategies, and the optimization of maternal and infant health outcomes. The review also identifies significant gaps in current knowledge and cautiously proposes hypotheses to explore these mechanisms. While preliminary, this framework offers an original perspective that may guide future research and open new paths in the study of human lactation biology.

Hygiene doi: 10.3390/hygiene5040045

Authors: Maria Cláudia Cuzzullin Marcelo Ivander Andrade Wanderley Maria Eduarda Pérez-de-Oliveira Ana Gabriela Costa Normando Anna Luíza Damaceno Araújo Joab Cabral Ramos Thaís Bianca Brandão Joel B. Epstein Alan Roger Santos-Silva Ana Carolina Prado-Ribeiro

Oral hygiene protocols and basic oral care (BOC) are paramount for patients undergoing cancer treatment, although based upon limited evidence from clinical trials. There are a range of oral care guidelines proposed in the literature mainly focusing on oral mucositis prevention. This systematic review aimed to define the best procedures and practices in the scope of oral hygiene protocols and BOC interventions for the prevention and/or treatment of oral complications in patients under cancer treatment. This systematic review was conducted according to the PRISMA guidelines, and the methods were previously established and registered in PROSPERO (CRD42022319455). A systematic search was completed in six databases (PubMed, Scopus, Web of Science, LILACS, and the Cochrane Library, and gray literature). Eleven studies were included in the qualitative analysis including non-randomized (n = 1) and randomized clinical trials (n = 6), and cohort studies (n = 4). The included studies showed the effectiveness of soft brushes for daily oral hygiene with topical fluoride, and the use of mouthwashes with 0.12% chlorhexidine, doxepin, and benzydamine and saline solutions, as well as dental scaling/prophylaxis in decreasing the severity of oral toxicities. Although not well-explored by well-designed clinical studies, BOC protocols may be effective in prevention oral toxicities from cancer therapy and are supported in multiple guidelines. This review synthesized available evidence on BOC in head and neck cancer patients, focusing on the effectiveness of hospital-based dental protocols during cancer treatment.

Hygiene doi: 10.3390/hygiene5040044

Authors: Dimitris Karaferis Dimitra Balaska Maria Eleni Karaferi Yannis Pollalis

Artificial intelligence (AI) is progressively being utilized in the healthcare sector to enhance efficiency, alleviate administrative burdens, and improve patient care outcomes. In the secondary healthcare sector, AI presents a range of opportunities as well as challenges. This study investigates the viewpoints of healthcare professionals regarding the adoption of AI in Greece, emphasizing the anticipated advantages and apprehensions associated with its integration. A cross-sectional descriptive study was carried out to collect responses from healthcare professionals at the General Hospital of “Evangelismos”, which is the largest hospital in Athens, Greece. A questionnaire was utilized and distributed over a period of four months, involving 513 registered healthcare professionals (comprising 136 physicians, 235 nursing staff, and 142 other healthcare personnel). Each participant had a minimum of one year of clinical experience and was selected using a convenience sampling method. The questionnaire comprised two parts: one focused on evaluating the AI knowledge and attitudes of healthcare professionals, and the other collected demographic data. The overall comprehension of knowledge pertaining to AI among healthcare professionals was evaluated as moderate, resulting in a mean score of 3.39. A distinction exists among different personnel categories, with physicians (M = 3.73) demonstrating a greater understanding of AI and a firm conviction that AI cannot supplant human positions. Conversely, nursing personnel appear to express apprehension regarding the implications of AI on the human experience, with a notable concern about potential replacement and job loss (M = 2.63), which was identified as the lowest-ranked issue. This latter concern is also echoed by other healthcare personnel (M = 2.90). Nevertheless, the majority of participants regard the prospective use of AI favorably, demonstrate confidence in its application, and contend that the benefits outweigh the possible risks. Sufficient training and ongoing updates would enhance employees’ comprehension of AI and their awareness of its potential benefits within the healthcare sector.

Hygiene doi: 10.3390/hygiene5040043

Authors: Arturo Juárez-García César Merino-Soto Javier García-Rivas

This study aimed to explore psychosocial factors in a sample of Peruvian workers, examine their convergence with the PROPSIT model, and identify the emergence of new or idiosyncratic psychosocial dimensions. At the same time, the quality and efficiency of the categorization process were evaluated. n = 48 workers were contacted by a non-probabilistic sampling method and asked to fill out a form with open-ended questions that explored negative stressors and positive engaging factors. Some strategies were used to assess the quality and efficiency of the categorization process. The results showed that the quality, speed, and reliability of the categorization procedure were satisfactory, and several categories were aligned with the PROPSIT model and other literature, both in their negative aspects (workload and rhythm, working hours, shifts, etc.) and positive aspects (rewarding tasks, atmosphere of unity, etc.). The emerging new categories were confined to aspects of teamwork and conflict climate, as well as topics such as order, cleanliness, and recreation. These findings underline the need to adapt existing models and instruments to capture idiosyncratic aspects of the Peruvian work environment. In conclusion, this study validated an efficient mixed approach for categorizing psychosocial work factors in Peru, revealing both PROPSIT-aligned and novel context-specific categories, and highlighting the need for culturally adapted tools and broader validation.

Hygiene doi: 10.3390/hygiene5030042

Authors: Gavin Banning Cindy Kim Carter Wilkerson Shelley J. Williams Karl Kingsley Victoria Sullivan

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.

Hygiene doi: 10.3390/hygiene5030041

Authors: Caroline Le Sénéchal Yoann Fautras Caroline Tokarski Sébastien Vilain

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.

Hygiene doi: 10.3390/hygiene5030040

Authors: Christian Paul Pablo Sanz Ruiz Muhamed Zeneli Klaus-Dieter Kühn

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.

Hygiene doi: 10.3390/hygiene5030039

Authors: Louis Alglave Manon Caudron Karine Faure Charlotte Moreau Catherine J. Mullié

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.

Hygiene doi: 10.3390/hygiene5030038

Authors: Angelo Michele Inchingolo Grazia Marinelli Valeria Colonna Benito Francesco Pio Pennacchio Roberto Vito Giorgio Francesco Inchingolo Daniela Di Venere Andrea Palermo Giuseppe Minervini Alessio Danilo Inchingolo Gianna Dipalma

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.

Hygiene doi: 10.3390/hygiene5030037

Authors: Riccardo Fontana Mattia Buratto Anna Caproni Chiara Nordi Mariangela Pappadà Martina Facchini Cesare Buffone Beatrice Bandera Luciano Vogli Peggy Marconi

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.

Hygiene doi: 10.3390/hygiene5030036

Authors: Vinícius do Amaral Flores Angélica Olivier Bernardi Bruna Lago Tagliapietra Maximiliano Escalona Maritiele Naissinger da Silva Juliana Copetti Fracari Marina Venturini Copetti Neila Silvia Pereira dos Santos Richards

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.

Hygiene doi: 10.3390/hygiene5030035

Authors: Paula Rocha Stephanie Norotiana Andriamiharisoa Ana Catarina Godinho Pierana Gabriel Randaoharison Lugie Harimalala Lova Narindra Randriamanantsoa Oni Zo Andriamalala Emmanuel Guy Raoelison Jane Rogathi Paulo Kidayi Christina Mtuya Rose Laisser Eyeshope J. Dausen Pascalina Nzelu Barbara Czech-Szczapa Edyta Cudak-Kasprzak Marlena Szewczyczak João Graveto Pedro Parreira Sofia Ortet M. Rosário Pinto

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.

Hygiene doi: 10.3390/hygiene5030034

Authors: Nikolas Lisboa Coda Dias Pedro Henrique Santos Serafim Ferraz Rayssa Lopes de Souza Mariana Felix Maccari Manoel Reverendo Vidal Wallisen Tadashi Hattori Stefan Vilges de Oliveira

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.

Hygiene doi: 10.3390/hygiene5030033

Authors: Konstantin Gus Kousoulas Ojasvi Dutta Harikrishnan Mohan Agustin Fernandez Santana

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.

Hygiene doi: 10.3390/hygiene5030032

Authors: Susana Carbajal-Ocaña Kristeel Ximena Franco-Gómez Valeria Atehortúa-Benítez Daniela Mendoza-Lozano Luis Vicente Prado-Cervantes Luis J. Melgoza-Ramírez Miguel Delgado-Rodríguez Mariana E. Elizondo-García Jorge Membrillo-Hernández

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.

Hygiene doi: 10.3390/hygiene5030031

Authors: Eun-Jeong Kim Su-Min Kang Min-Jeong Ko Da-Yeon Kim Je-Yeong Kim Mi-Seong Choi Hye-ju Lee

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.

Hygiene doi: 10.3390/hygiene5030030

Authors: Magdalini Christodoulou Ourania S. Kotsiou Konstantinos Tsaras Charalambos Billinis Konstantinos I. Gourgoulianis Dimitrios Papagiannis

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.

Hygiene doi: 10.3390/hygiene5030029

Authors: Angeliki Flokou Styliani Spyrou Dimitris A. Niakas Vassilis Aletras

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.

Hygiene doi: 10.3390/hygiene5030028

Authors: Maria Teresa Montagna Matteo Moro Beatrice Casini Ida Iolanda Mura Gianfranco Finzi Valentina Spagnuolo Antonella Francesca Savino Fabrizio Fasano Francesco Triggiano Lucia Bonadonna Osvalda De Giglio

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.

Hygiene doi: 10.3390/hygiene5030027

Authors: Charles P. Gerba Stephanie A. Boone Julie McKinney M. Khalid Ijaz

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.

Hygiene doi: 10.3390/hygiene5030026

Authors: Susy P. Saraiva Elaine C. Marqueze Claudia R. C. Moreno

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.

Hygiene doi: 10.3390/hygiene5020025

Authors: Eun-Ha Jung Mi-Kyoung Jun

Malocclusion can arise due to various causes and may impact not only temporomandibular joint (TMJ) disorders but also overall systemic health. This study aimed to evaluate the association of individual occlusal characteristics with sleep quality and stress levels in female college students. Occlusal analysis was conducted using T-Scan III on 84 participants (age: 21.7 ± 2.09), while sleep quality and stress levels were assessed using the Korean version of the Pittsburgh Sleep Quality Index and self-reported stress questionnaires, respectively. Sleep quality was categorized as “good sleeper” (≤5 points) or “poor sleeper” (>5 points). Stress levels were classified as Normal (≤13), Initial stress (14–16), Moderate stress (17–18), or Severe stress (≥19). The results indicated no significant differences in sleep quality based on occlusal characteristics. However, participants with Type A occlusion (posterior contact without anterior contact) exhibited higher stress scores than those with Type B occlusion. Correlation analysis showed a weak negative association between stress scores and occlusion type, but no significant relationship was found between occlusal characteristics, sleep quality, and stress levels. In conclusion, individual occlusal characteristics have limited explanatory power regarding their effects on sleep quality and stress.

Hygiene doi: 10.3390/hygiene5020024

Authors: Maria Antoniadou Sofia Sokratous Evangelos Dimitriou Ioannis Tzoutzas

Background: Infection control is fundamental in dental practice, especially following the COVID-19 pandemic, which highlighted the variability in students’ adherence to disinfection protocols. This study aimed to evaluate the knowledge, attitudes, and practices of the fifth-year dental students at the National and Kapodistrian University of Athens regarding antisepsis and infection control, and to assess the effectiveness of an educational intervention. Methods: A pre-post interventional study was conducted involving two in-person seminars, supplementary e-learning material, and a structured questionnaire administered before and after the intervention. The survey assessed the knowledge, clinical practices, and attitudes toward infection control, including vaccination history and prior exposure incidents. Results: The intervention led to statistically significant improvements in infection control knowledge, especially in risk-based sterilization strategies, disinfectant classification, and PPE use. Students with prior hepatitis B vaccinations and antibody testing demonstrated higher baseline scores and more significant knowledge gains. However, some misconceptions, particularly regarding surface disinfection and prosthetic care, persisted after the intervention. Conclusions: The findings support the effectiveness of structured educational interventions in improving infection control awareness among dental students. Practical, simulation-based training and earlier curriculum integration are recommended to enhance compliance and ensure safe clinical practice.

Hygiene doi: 10.3390/hygiene5020023

Authors: Miltiadis A. Makrygiannakis Angeliki Anna Gkinosati Sotirios Kalfas Eleftherios G. Kaklamanos

Fluoride mouthwashes are often recommended by dental professionals due to their proven benefits for oral hygiene. However, it is vital to acknowledge that these products may have undesirable effects on orthodontic treatment outcomes, particularly by altering the biomechanical properties of orthodontic devices and their components. To gain a comprehensive understanding of this potential issue, an extensive and systematic search was conducted across seven distinct databases. PRISMA extension for scoping reviews (PRISMA ScR) guidelines were followed. Following a detailed evaluation and careful scrutiny of the available evidence, a total of seven relevant studies met the inclusion criteria and were incorporated into the current scoping review. Findings indicated that regular intraoral use of fluoride-containing mouthwashes could lead to heightened corrosion and greater release of metal ions from stainless-steel brackets and nickel–titanium (NiTi) archwires. Additionally, the mechanical properties and structural integrity of titanium–molybdenum alloy (TMA) wires were negatively influenced by exposure to fluoride mouthwashes. Although existing evidence highlights these potential drawbacks, there remains a clear necessity for additional comprehensive research. Given the possibility that fluoride mouthwashes could adversely influence orthodontic treatment effectiveness, orthodontists and dental clinicians must exercise cautious judgment and deliberate consideration when prescribing fluoride-based mouthwashes for patients undergoing orthodontic therapy.

Hygiene doi: 10.3390/hygiene5020022

Authors: Stephanie A. Boone Nick D. Childress Norma Patricia Silva-Beltrán Julie McKinney M. Khalid Ijaz Charles P. Gerba

Restrooms are associated with the transmission of bacterial and viral illnesses. Disinfecting contaminated surfaces is associated with reducing transmission risk. The goal of this study was to determine how cleaning/disinfecting frequency affects restroom pathogen contamination. The Phase 1 intervention included cleaning toilet surfaces (the toilet bowl, water, and rim) using 9.5% w/w hydrochloric acid. The Phase 2 intervention used fomite-specific products to clean/disinfect additional restroom surfaces, including the vanity countertop, sink faucet handle, toilet seat, flush handle, floor, and doorknob. A designated household member was responsible for cleaning/disinfecting surfaces at the beginning of each interval. Fomite sample collection was randomized, and samples were tested for heterotrophic, coliform, and Escherichia coli bacteria after specified intervals: 1, 2, 3, or 7 days. The greatest numbers of bacteria and largest reductions occurred on fomites after three days. A statistically significant difference was found for heterotrophic bacteria (p = 0.009), coliforms (p = 0.10), and E. coli (p = 0.13) with cleaning/disinfecting every three days. A quantitative microbial risk assessment (QMRA) estimated a >98% reduction in risk of infection by norovirus with an every-3-day cleaning/disinfection routine on the most heavily contaminated sites. Results indicate an optimal cleaning frequency of twice weekly for minimizing exposure to pathogens.

Hygiene doi: 10.3390/hygiene5020021

Authors: Beatrice Pattaro Michele Dolcini Andrea Brambilla Stefano Capolongo

Background: This study examines the role of the built environment in mitigating risk in healthcare facilities, with a particular focus on how the design of hospital infrastructures can influence and improve the safety of patients, staff, and visitors. Methods: A two-phase mixed-methods approach was adopted. First, a scoping literature review was conducted to identify design-based strategies targeting five categories of risk: healthcare-associated infections (HAIs), indoor air quality (IAQ), safety, falls, and emergency resilience. Based on this review, a structured questionnaire was developed and administered to a sample of hospital facilities in Northern Italy to assess the implementation of the strategies emerged. Results: The literature review identifies recurring specific design solutions and strategies that have proven effective in mitigating risks in healthcare infrastructures in the following dimensions: infection mitigation, indoor air quality, falls reduction, safety, emergency preparedness. At the same time, survey data from (n = 9) hospitals indicate a significant implementation gap. Key shortcomings included a lack of spatial flexibility, limited environmental monitoring (especially for IAQ and acoustic conditions), and underutilization of antibacterial surfaces. Antibacterial flooring and wall finishes were absent in (n = 4/9) and (n = 6/9) of the facilities, respectively. IAQ monitoring was mostly confined to surgical areas, with (n = 0/9) facility reporting comprehensive building-wide monitoring. Only two (n = 2) facilities reported adaptable spaces suitable for emergency conversion and accessible green areas. Conclusions: This study provides a comprehensive overview of risk mitigation strategies in hospital design. The results reveal critical gaps in implementation, particularly in spatial flexibility, environmental monitoring, and antimicrobial surfaces. Future research should focus on developing adaptable design models that are context-sensitive, scalable, and capable of enhancing healthcare resilience in response to emerging global health threats.

Hygiene doi: 10.3390/hygiene5020020

Authors: Dimitris Charalambos Karaferis Dimitris A. Niakas

Patients represent the primary stakeholders in health systems, making it crucial to enhance their level of satisfaction. The relationship between physician and patient is crucial during treatment, especially for patients dealing with long-term issues. Generally, every illness has the potential to cause high levels of stress, leading to varying reactions from individuals, influenced by factors such as age, personality, relationships with caregivers, treatment environment, and specific disease characteristics. As the healthcare provider acquires a more profound insight into how the disease affects the patient’s quality of life, the relationship between the patient and the physician evolves. In this study, we analyzed information from hospital clinics that took part in the survey for the same timeframe, from July 2019 to December 2020. Correlated data were derived from two separate surveys. The initial survey, created by Spector, focused on gathering feedback from health professionals in hospitals, with 3278 responses collected. A second survey was designed to gather feedback from patients in hospitals, with 2851 responses collected. The survey revealed a total of 97 common observations among physicians and patients in the hospital clinics. According to the findings from the two-stage least squares (2SLS) regression analysis, an essential method for researchers addressing endogeneity challenges in structural equation modeling, an increase of 1 point in the job satisfaction of physicians is associated with a corresponding positive increase of 1.12 points in patient satisfaction. At the heart of effective healthcare is the relationship between physicians and patients. Positive attitudes from patients during medical appointments help improve health outcomes, especially for those with long-term illnesses. In order to encourage patients to take an active role, physicians need to focus on enhancing the physician-patient bond, humaneness, creating trust and stable communication, encouraging the sharing of information, and empowering patients to participate in decision-making.

Hygiene doi: 10.3390/hygiene5020019

Authors: Alba Soledad Aquino-Domínguez Melisa Gómez-López Sergio Roberto Aguilar-Ruiz

Breast milk is vital for infant survival, protecting against infections and strengthening the immune system. In addition to nutrients, breast milk contains beneficial microorganisms, antimicrobial peptides and proteins (APPs), including lactoferrin and lysozyme, and peptides such as defensins and cathelicidins that destroy harmful bacteria and regulate the neonatal immune response. Breast milk also promotes the growth of beneficial gut bacteria (Bacteroidaceae and Bifidobacteriaceae) while reducing harmful pathogens, fostering a healthy gut microbiome, and supporting long-term infant health. Traditionally, research on antimicrobial proteins and milk microbiota has been conducted in isolation. However, at the molecular level, these components do not function independently; they interact synergistically, influencing immunomodulation, inflammation, and the composition of the gut microbiome. Therefore, this review aims to provide an overview of the discovery and identification of APPs in breast milk, the dynamic relationship between the breast milk microbiota, and the potentiation of artificial feeding with supplemented formulas when breastfeeding is impossible, benefits on newborn immune systems, and even the benefits to breast tissue.

Hygiene doi: 10.3390/hygiene5020018

Authors: Silvia De Gaetano Elena Ponzo Angelina Midiri Giuseppe Mancuso Daniele Filippone Giovanni Infortuna Sebastiana Zummo Carmelo Biondo

In recent decades, the world has observed the emergence and re-emergence of a multitude of previously non-existent or re-emerging infectious diseases, for which there is a paucity of timely and effective preventative measures. The WHO has published a catalogue of priority pathogens that are likely to trigger future epidemics, with the objective of designing effective prophylactic and therapeutic interventions. The rationale behind these interventions is rooted in a comprehensive understanding of the aetiology, epidemiology, and pathogenesis of the target diseases. While it is imperative to acknowledge the pivotal role that evolutionary changes in pathogens play, it is equally important to recognise the influence of a multifaceted interplay of factors on the emergence and re-emergence of infectious diseases. These include changes in human populations, the vectors and reservoirs of exposure, and environmental changes. This review summarises the aetiology, epidemiology, and pathogenesis of the ten WHO priority diseases, as well as those with high epidemic potential that are already the focus of specific control programme initiatives, such as HIV/AIDS, tuberculosis, and malaria diseases. Furthermore, this review concentrated on the means of addressing these infections through public health surveillance and response systems. Such systems must be designed to rapidly detect unusual and unexpected disease patterns, track and share information in real time, and rapidly mobilise global responses, which are the most important ways to effectively contain transmission.

Hygiene doi: 10.3390/hygiene5020017

Authors: Karan Varshney Pavan Shet Mariyah Hoosenally

The spread of diarrheal disease is a major public health problem, and a lack of proper sanitation is an important contributor to this. In 2014, the Swachh Bharat Mission (SBM) was initiated by the Government of Bharat (India) to improve sanitation across the entirety of the country. SBM has received worldwide praise for its success, and it is now being expanded further. For the purposes of guiding the ongoing implementation of SBM and determining where it has or has not been successful, the objective of this scoping review was to determine the perceptions, knowledge, and attitudes of individuals regarding SBM. After screening 5454 research articles across seven databases, nine total articles were eligible for inclusion in this review. In spite of the reduced number of selected studies, numerous key themes emerged: Many chose to greatly praise SBM for its past successes; individuals felt that mass media should continue to be used to raise awareness; mindsets and a lack of awareness continue to serve as barriers to change; addressing inadequate waste disposal and waste segregation should be a very high priority to ensure cleanliness in Bharat. The findings of this review can guide policymakers, government workers, non-governmental organizations, and researchers in ensuring the continual development of SBM in its second phase and beyond the successful achievement of its aims, though the inherently limited generalizability of the qualitative studies included in this scoping review should also be recognized. Further research that is both qualitative and quantitative in nature is needed to develop a deeper understanding of the perceptions regarding SBM and the ways in which the ongoing program can be improved.

Hygiene doi: 10.3390/hygiene5020016

Authors: Mirella Di Dio Marco Santulli Mariangela Pagano Anna Maria Rossi Renato Liguori Giorgio Liguori Valeria Di Onofrio

Legionella bacterium has the aquatic environment as its natural reservoir. In humans, it can cause a form of interstitial pneumonia called legionellosis which can be transmitted by inhalation of contaminated water aerosols. Legionella infection occurs more frequently in certain more susceptible population groups, including smokers, alcoholics, men, the elderly, as well as people with acquired immunodeficiency syndrome, hematological cancers, and diabetes mellitus. This study aimed to evaluate the effectiveness of the new Italian National Guidelines for the prevention of Legionella colonization in water systems application by analyzing the environmental monitoring data of Legionella carried out in healthcare facilities in the Campania region from 2019 to 2022. The secondary objectives were to estimate the most observed serogroups of L. pneumophila and to analyze the possible link between water temperature and the presence of Legionella, respectively. From our data, it emerged that in 2019, 41.1% of the examined facilities were contaminated by the Legionella genus; in 2020, the contamination percentage was 42.9%; in 2021, it was 54.5%; in 2022, it was 45.5%. Instead, the Legionella positivity rate decreased from 2019 (54.3%) to 2022 (52.4%), suggesting a possible positive influence of more restrictive prevention and control measures. The prevalent species was Legionella pneumophila, particularly serogroup 1; water temperature was the risk factor implicated in Legionella contamination.

Hygiene doi: 10.3390/hygiene5020015

Authors: Muhammad Yasir Mark D. P. Willcox John Ings Peter van Bruinessen

Background: Respiratory viruses spread through airborne droplets and aerosols, causing highly contagious acute respiratory syndromes in humans. This study evaluated the antiviral potential of vapours of catmint-oil-based formulations against respiratory viruses. Methods: The antiviral activity of formulations with or without catmint oil (CO) in solution or in aerosolised form was determined against influenza virus H1N1 ATCC VR-1469 and mouse hepatitis virus (MHV-1) ATCC/VR261. In solution, both viruses were exposed to CO formulations for 2–3 h. In aerosolised form, H1N1 was exposed to formulations for 2 min in a closed cylinder and MHV-1 for 10 min in a booth. The antiviral effect of the formulations was evaluated by growing H1N1 in a Madin–Darby canine kidney (MDCK; ATCC-CRL-2936) and MHV-1 in A9 ATCC/CCL 1.4 cells using TCID50 and a plaque assay, respectively. Transmission electron microscopy (TEM) was conducted to investigate the mode of action of the formulations. Results: In solution, the formulation containing hydrogenated CO (HCO), bromelain, N-acetylcysteine and Tween 20 (Formulation (1)) reduced the viability of H1N1 by 2.6 ± 0.07 log10 (p = 0.025) and MHV-1 by 4.5 ± 0.14 log10 (p = 0.014) within 2–3 h. In vapourised form, Formulation (1) produced similar antiviral effects against H1N1, reducing it by 3.00 ± 0.07 log10 (p = 0.002) within 2 min, and Formulation (1) produced a 3.00 ± 0.07 log10 reduction of MHV-1 (p < 0.001) within 10 min (the minimum time needed to detect infective viral particles in the experimental set-ups). Formulation (3) (without bromelain) reduced H1N1 by 1.57 ± 0.14 log10 (p = 0.008) after 2 min and MHV-1 by 1.3 ± 0.04 log10 (p = 0.057) after 10 min. In the absence of catmint oil (Formulation (4)) or in the absence of catmint oil and bromelain (Formulation (5)), there were only slight reductions in the viability of aerosolised H1N1 (1.00 ± 0.14 log10, p = 0.046; <1 log10, p = 0.966, respectively) and MHV-1 (1.07 ± 0.02 log10, p = 0.013; 0.16 ± 0.03 log10, p = 0.910, respectively). The TEM analysis showed that the formulation disrupted the H1N1 envelopes and caused a reduction in size of the viral particles. Conclusions: The catmint-oil-based formulations reduced the H1N1 and MHV-1 by disrupting the vial envelopes.

Hygiene doi: 10.3390/hygiene5020014

Authors: Md S. Zaman S. M. Golam Alam Mohammed S. Razzaque

The human oral microbiome plays a vital role in maintaining oral and systemic health. This diverse microbial community includes over 700 bacterial species, some of which are implicated in developing systemic diseases, particularly cardiovascular diseases (CVDs). Research highlights a strong association between periodontal disease and increased cardiovascular risk, suggesting that good oral hygiene practices may reduce the incidence of CVDs. Porphyromonas gingivalis and Fusobacterium nucleatum drive chronic inflammation in periodontal disease; these bacteria can extend beyond the mouth and contribute to systemic inflammatory responses. The inflammatory factors, including C-reactive protein (CRP), interleukins (IL-1, IL-6), and tumor necrosis factor-alpha (TNF-α), damage blood vessels, impair endothelial functions, and promote atherosclerosis, all key events in CVD progression. Additionally, oral pathogens may accelerate plaque formation in arteries, increasing the risk of ischemic heart and brain diseases. Studies show a 28% increased risk of heart disease in individuals with periodontal disease. Treating periodontal disease can improve endothelial function and reduce inflammatory markers, emphasizing oral health management as a potential preventive strategy for CVD. Public health initiatives that emphasize oral hygiene and early periodontal disease treatment are crucial for broader cardiovascular care.

Hygiene doi: 10.3390/hygiene5020013

Authors: Lidia Caporossi Simona Di Renzi Mariangela De Rosa Silvia Capanna Elisa Partenzi Iacopo D’Alessandro Bruno Papaleo

Background: Lead compounds are chemicals of high toxicological concern and are suspected to interact with the thyroid axis. Method: A cohort study was carried out involving 70 workers from a petrochemical company exposed to inorganic lead compounds. All recruited workers were given a clinical anamnestic questionnaire aimed at characterizing their endocrine and thyroid status. A blood test was conducted to dose the amount of lead, thyroid hormones (FT3, FT4, TSH), and antibodies (TGAb and TPOAb). Samples were stratified according to working seniority and lead exposure levels. A regression study was conducted to highlight trends in hormones and antibodies versus lead levels. Results: Most of the dosages are within the normal ranges. The regression study showed how higher lead values are correlated with a reduction in TSH and an increase in FT3 and FT4. There is a statistically significant increase in TPOAb in the most exposed workers. Conclusions: The trends of thyroid hormones may suggest a tendency towards hyperthyroidism for higher lead exposure, while the increase in TPOAb could indicate a greater predisposition to the development of autoimmune thyroid diseases.

Hygiene doi: 10.3390/hygiene5010012

Authors: Muriel Schulze Franziska Nankemann Anne Tellen Julia Nitz Yanchao Zhang Stefanie Leimbach Volker Krömker

Intermediate cluster disinfection plays a vital role in preventing the transmission of mastitis pathogens during milking. This study evaluates the efficacy of different disinfection methods on teat liners through a randomized controlled field trial conducted on three dairy farms in Germany. The treatments assessed included dipping the liners in water, peracetic acid solution (PAS, 0.1%), and plasma-activated buffered solution (PABS). Total bacterial count (TBC) and the bacterial load of presumptive Staphylococcus (S.) aureus were measured using the wet–dry swab method. The results showed that PAS significantly reduced both TBC and S. aureus by 90% and 99%, respectively. PABS also demonstrated a significant reduction in the bacterial load of S. aureus. In contrast, dipping in water had no significant effect on either TBC or S. aureus bacterial counts. This study suggests that while PAS is the more effective disinfectant, PABS may offer an alternative with some antimicrobial activity. Further research on intermediate cluster disinfection in general is necessary to optimize its application and assess its long-term effectiveness in dairy farming practices.

Hygiene doi: 10.3390/hygiene5010011

Authors: Michael Pearson Samuel Stewart Linda Ma Karl Kingsley Victoria Sullivan

Endocyn is a root canal irrigant with a stable formulation of hypochlorous acid (HOCl), and should have significant antimicrobial effects. However, there are no available studies that evaluate these effects on different types of bacterial species. In this prospective in vitro study, bacterial species were grown with and without the addition of Endocyn to measure the effects on the Gram-positive bacteria Actinomyces naeslundii, Lactobacillus acidophilus, Streptococcus gordonii, and Streptococcus mutans, as well as the Gram-negative bacteria Porphyromonas gingivalis, Fusobacterium nucleatum, and Veillonella parvula. Turbidity was measured at 24 h, and the differences between the baseline and experimental treatments were measured using two-tailed Student’s t-tests and verified using ANOVA. Gram-positive bacteria were inhibited in the range of −8.2% to −35.5%, p = 0.14 to p = 0.004, while Gram-negative bacteria were inhibited in the range of −16.7% to −41.4%, p = 0.04 to p = 0.001, which were similar to the effects of 5% NaOCl (bleach). These data demonstrated that Gram-positive bacteria were somewhat resistant to Endocyn at lower levels but were inhibited at all higher concentrations, while Gram-negative bacteria were susceptible to Endocyn at all levels, and increased at higher concentrations. These results provide clinically relevant data regarding the efficacy of this disinfectant against common oral pathogens (and commensal bacteria), and are important as they provide evidence regarding public health and the environmental safety of clinical protocols regarding endodontic hygiene.

Hygiene doi: 10.3390/hygiene5010010

Authors: Saverio Cosola Sandy Manuela Fanelli Antonia Sinesi Roberta Lasala Giacomo Oldoini Annamaria Genovesi Andrea Butera Andrea Scribante

The literature has identified several common acute and chronic complications associated with periodontal health during pregnancy, such as gingivitis, recession, periodontitis, and changes in systemic health, such as gestational diabetes, which may act as additional risk factors for chronic periodontal disease. Are the general public and health care providers aware of the potential risks of systemic and intrauterine inflammation caused by bacteria from the oral microbiota entering the bloodstream through inflamed gingival tissue and affecting the fetoplacental unit, leading to complications, such as preterm birth and reduced prenatal growth? A questionnaire-based survey, conducted between January 2023 and June 2023, aimed to assess patients’ personal oral hygiene practices and their understanding of the link between pregnancy and periodontal health. A total of 826 people completed a questionnaire for this study. The majority of women surveyed (86.9%) were not pregnant, but 77.7% had at least one child. The majority of women believed in good oral hygiene during pregnancy but lacked knowledge about how oral care can affect pregnancy outcomes.

Hygiene doi: 10.3390/hygiene5010009

Authors: Carlos Braga Elken Gomes Rivaldo Arthur Saavedra de Paula Rim Bourgi Louis Hardan Naji Kharouf Mohammad Qaddomi Youssef Haikel Celso Afonso Klein-Junior

The customization and handling of implant abutments in prosthetic laboratories can lead to microbial contamination, requiring disinfection before clinical use. This study evaluated cross-contamination in abutments from three prosthetic laboratories, identifying microorganisms and assessing the disinfection efficacy of 2% chlorhexidine digluconate (2% CL) and ultraviolet C (UV-C) radiation. Sixty abutments were analyzed, with contamination detected in 83%, predominantly Enterococcus faecalis (34.2%) and Staphylococcus epidermidis (17.8%). Post-disinfection, CFU reduction was 92% (UV-C) and 93% (CL), confirmed by the Wilcoxon test (Z = −4.373; p < 0.001). A comparative analysis using the Kruskal–Wallis test showed no significant difference between the two methods (p > 0.05). These findings confirm that both UV-C and CL effectively reduce microbial load, providing reliable disinfection protocols for clinical practice.

Hygiene doi: 10.3390/hygiene5010008

Authors: Rachel Lau Jason Kwan Kimberley Marks-Beaubrun Ruben Cudiamat Min Qun Ellen Chen Krista Orejana Filip Ralevski Andrea K. Boggild

Background: Microscopy is the conventional method for the identification of gastrointestinal parasitic pathogens in fecal specimens; however, it presents numerous challenges, including high technical expertise burden, multiple staining procedures, and prolonged turnaround time. Molecular methods provide higher throughput and potentially higher sensitivity and specificity. Methods: We validated a commercial, automated DNA extraction platform and multiplex parasitic real-time PCR panel (Seegene AllplexTM GI-Parasite Assay) detecting six protozoal pathogens: Blastocystis hominis (Bh), Cryptosporidium spp., Cyclospora cayetanensis (Cc), Dientamoeba fragilis (Df), Entamoeba histolytica (Eh), and Giardia lamblia (Gl) in unpreserved fecal specimens submitted for diagnostic parasitology. Microscopy was the reference standard for all organisms, with stool ELISA as an additional reference assay for Eh. Results: Among 461 unpreserved fecal specimens, sensitivity, specificity, positive predictive and negative predictive values of the enteric multiplex for fresh specimens were as follows: 93%, 98.3%, 85.1%, 99.3% for Bh; 100% for all measures in Cryptosporidium and Cc; 100%, 99.3%, 88.5%, 100% for Df; 33.3%, 100%, 100%, 99.6% for Eh; and 100%, 98.9%, 68.8%, 100% for Gl, respectively. With the addition of 17 frozen specimens, the sensitivity for Eh increased to 75%. On a per-batch basis, the molecular platform reduced pre-analytical and analytical testing turnaround time by 7 h. Conclusions: The enteric multiplex platform provides a useful diagnostic tool for clinically relevant enteric protozoa, including Cryptosporidium spp., Cyclospora cayetanensis, Dientamoeba fragilis, and Giardia lamblia. Further evaluation of the assay is required for Entamoeba histolytica prior to clinical use; however, given the widespread availability of confirmatory serology and stool antigen testing for E. histolytica, such performance limitations are of lesser concern.

Hygiene doi: 10.3390/hygiene5010007

Authors: Miguel Antonio Fudolig Robert E. Davis Kavita Batra Manoj Sharma

Maintaining proper hand hygiene is crucial in preventing the spread of infections and other communicable diseases. It is imperative to determine the factors that affect the likelihood of initiating and maintaining the recommended handwashing behavior, especially during a pandemic. This quantitative, secondary study employed a latent profile analysis (LPA) to identify the different attitudes toward behavior change based on the Multi-Theory Model (MTM) framework in the context of following the guidelines provided by the Centers for Disease Control and Prevention (CDC) during the COVID-19 pandemic. Data were collected from 602 college students at a large university in the southern region of the United States (U.S.) in 2020. Seven distinct profiles were identified, each reflecting unique attitudes toward following the recommended handwashing guidelines. Age (p < 0.01) and gender (p < 0.01) disparities were observed between profiles. This study is the first to apply LPA within the MTM framework and provides new insights for the development of targeted interventions based on the construct score profile.

Hygiene doi: 10.3390/hygiene5010006

Authors: Philippe Hartemann Antoine Montiel

Throughout history, humans have sought to drink water that is good for their health, according to the knowledge of the time. Hippocrates’ definition of water quality, “good water should be clear, light, aerated, without any perceptible odor or taste, warm in winter and cold in summer”, remained virtually unchanged until 1887, when it was added that water should dissolve soap and foam well, be clear and colorless, have a pleasant taste, leave no large deposits after boiling, and cook vegetables and wash clothes well. This definition guided all treatments to remove the substances responsible for cloudiness, odor and discoloration, as well as the choice of resources: clear water and water with low mineral content. The discoveries by Pasteur and Koch led to the addition of microbiological criteria, like the absence of pathogens, and the definition of microbiological indicators. Throughout the 20th century, advances in scientific knowledge in microbiology, chemistry and toxicology led to major progress in treatment methods. These filtration and disinfection treatments are described here according to their historical implementation. Due to progress in numerous areas, e.g., both chemical and microbiological analytical detection limits, speed of information flow and origins of certain diseases that are discovered to be waterborne, the consumer is now exposed to anxiety-provoking news (microplastics, eternal pollutants (cf. per- and polyfluoroalkyl substances (PFASs)), drugs, pesticides residues, etc.). Thus, the consumer tends to lose confidence in tap or bottled water and turn to buying home purifiers. Drinking water treatment will continue to evolve with more sophisticated processes, as analytical progress enables us to expect further developments.

Hygiene doi: 10.3390/hygiene5010005

Authors: Vanshika Maini Rupanjan Roy Gargi Gandhi Aditi Chopra Subraya G. Bhat

Artificial intelligence (AI) is one of the most promising technological advancements that have revolutionized the healthcare sector (medicine and dentistry). AI and its subsets, such as machine learning (ML), artificial neural networks (ANNs), and deep learning (DL), are being used in dentistry for data recording and management, patient education, radiographic interpretation, diagnosis, and treatment plans. AI and ML tools are commonly employed to improve oral hygiene and patient compliance. This narrative review paper discusses the innovations in AI-based plaque control aids (toothbrushes and interdental aids) that have improved overall health and patients’ hygiene compliance. We performed a literature search using different databases using the following keywords: “Artificial intelligence or machine learning or robots or robotics” AND “Toothbrush OR Smart toothbrush”. We included all the studies evaluating the use of any smart toothbrush, AI, or robotics for oral hygiene, plaque control, and patient education. AI-based smart toothbrushes helped patients to brush effectively by indicating the amount of pressure and the time taken for brushing, along with providing feedback on their brushing performance. Many microrobots can even recognize and automatically remove biofilm. Some AI-based smart toothbrushes are beneficial for children, patients with disabilities lack of manual dexterity, and neurological disorders. However, dental professionals choose AI-based smart toothbrushes for patients with poor oral hygiene and poor compliance for more effective control of oral diseases and to provide better health.

Hygiene doi: 10.3390/hygiene5010004

Authors: Vincenzo Marcotrigiano Alice Prete Chiara Canal Angela Padoin Erica Bino Igor Campigotto Alberto Lovat Flavia Campigotto Mattia Manzi Jacopo Fagherazzi Andrea Da Rech Anna Voltolini Nahuel Fiorito Pamela D’Incà Maria Parpinel Sandro Cinquetti

In its broadest sense, the mountain environment represents a setting useful for promoting health. This Italian project, conceived and applied in the Dolomites, aims to encourage multi-sector preventive public health policies; improve refuge hygiene; strengthen safety requirements; and implement projects aimed at promoting health. With the support of healthcare workers (HCWs) of the Prevention Department and other professionals in this field, 27 establishments in the Alpine territory have joined the project, benefiting from being recognized as a “Healthy and Safe Refuge” at the end of the quality improvement process. This process consisted of a documentary assessment, an on-site evaluation, and an assessment of analytical evidence relating to parameters investigated by both HCWs and refuge managers according to specific protocols. Data were collected from a specific database via an application to join the project and from sampling reports of parameters investigated for evaluating the quality of water intended for human consumption. The outdoor Alpine environment has recently witnessed extraordinary development and must continue to meet all health and safety requirements, especially in view of a global event—the Olympic Games—that will be hosted in the Dolomites area in 2026.

Hygiene doi: 10.3390/hygiene5010003

Authors: Anna Maria Spagnolo

Aspergillus spp. are ubiquitous fungi present in soil, organic debris, water, decaying vegetation and dust produced in renovation and/or building work. Several studies have shown the presence of aspergilli in various healthcare environments. Typically, thousands of fungal spores are inhaled every day, but if spore clearance fails (typically in immunocompromised patients), fungi can grow and invade lung tissue, causing invasive aspergillosis (IA) which is one of the most frequent infections in highly immunocompromised patients. Aspergillus fumigatus is the most common species involved; this species can be attributed to about 80% of the cases of aspergillosis. According to the WHO, Aspergillus fumigatus is one of four critical priority fungi. The first-line treatment of diseases caused by Aspergillus, in particular IA, is based on triazole antimycotics. Unfortunately, resistance to antimycotics is increasing, partly due to their widespread use in various areas, becoming a significant concern to clinicians who are charged with caring for patients at high risk of invasive mycoses. A recent WHO report emphasised the need for strategies to improve the response, and in particular strengthen laboratory capacity and surveillance, support investment in research and strengthen public health interventions for the prevention and control of fungal infections through a One Health approach.

Hygiene doi: 10.3390/hygiene5010002

Authors: Lilian Jeptoo Kiborus S. Travis Altheide Jason W. Marion

The spread of methicillin-resistant Staphylococcus aureus (MRSA) in community settings, including fitness/exercise centers, remains relevant for public health. MRSA, a cause of severe infections in some, can be transmitted through shared equipment and skin contact. Understanding its prevalence and the frequency of antibiotic resistance in such environments can be useful for informing hygiene and intervention strategies. For investigating, multiple environmental swabs were collected from 14 different sites within a fitness facility, including equipment and locker rooms. Samples were collected for characterizing the prevalence of staphylococci (including MRSA), E. coli, and carbapenem-resistant Enterobacterales (CRE). Isolated colonies were identified biochemically and evaluated for antibiotic resistance. Logistic regression was applied to assess risk across different surfaces. Among 42 samples, the highest prevalence of Staphylococcus spp. was on locker room surfaces. S. aureus was prevalent on locker room floors and benches. Non-S. aureus species, such as S. saprophyticus and S. haemolyticus, were common. Resistance to oxacillin and penicillin was widespread, particularly among non-S. aureus species. E. coli was detected once, and CRE was not detected. Fitness center surfaces can harbor staphylococci, including MRSA. The results obtained corroborate other studies finding notable antibiotic resistance among staphylococci from fitness center surfaces. Hygiene improvements, including personal hygiene actions, are essential for reducing transmission risks.

Hygiene doi: 10.3390/hygiene5010001

Authors: Carolina Marques Borges Matthew Conlan Areeb Khateeb Emma Tomczynski

The lack of social determinants of health has significantly influenced COVID-19 mortality; however, the few studies that have investigated the relationship among social capital, vulnerability, and COVID-19 mortality have either shown mixed directions of association or were not conducted at a larger scale on the state level. Our study aimed to fill this research gap. We aimed to test the association of comprehensive vulnerability and social capital measures and COVID-19 mortality in 50 states in the U.S. during the first year of the pandemic. We hypothesized that states with low social capital would register the highest mortality rates and high vulnerability would be proportional to a high number of deaths due to COVID-19 in the U.S. Methods: Our ecological analytic study used aggregate secondary data from nine open access databases. The outcome was COVID-19 mortality (January 2020 to July 2021). The main exposures were social capital and social vulnerability. We also tested 28 covariates and selected socioeconomic variables at the state level. Statistical analysis consisted of a multivariate technique factor analysis and a bivariate Local Indicators of Spatial Association (LISA) analysis. Findings: Social capital (SCI) and social vulnerability (SVI) explained 57% of the COVID-19 mortality rate variation (R2 adjusted = 0.57). This analysis resulted in a statistically significant model (p ≤ 0.001), where SCI (β = 23.256, t = 2.649, p = 0.011) and SVI (β = 150.316, t = 4.235, p = 0.001) were predictors for COVID-19 mortality in the investigated period.

Hygiene doi: 10.3390/hygiene4040040

Authors: Sueny Andrade Batista Emanuele Batistela dos Santos Gabriel Teles Câmara Ester Cardoso Paes Rose Pedro Tourinho Dantas Raquel Braz Assunção Botelho Verônica Cortez Ginani

This study assessed raw vegetable sanitizing in Brazilian schools and identified barriers to standards. This experimental and quantitative study was conducted in 12 school food services in the Federal District (Brazil) public primary education institutions. Microbiological analyses were conducted with vegetable samples (before and after sanitizing) and water used in the sanitization process, collected before the process. The Petrifilm®E. coli/Coliform Count Plates and COLIlert methods were used to evaluate vegetables and water samples, and a checklist of good practices was applied in each school food service to identify barriers to proper sanitization. Thirty-five samples of raw vegetables were offered to students, 32 samples of water, and 17 hygiene processes were evaluated. The results indicate that 76.5% (n = 13) of hygiene processes were considered unsatisfactory, with an average increase of 5.8 log CFU g−1 (DV = 7.4) in the initial microbial load in 47.1% (n = 8) of the evaluated processes; moreover, 33.3% (n = 6) of the samples exceeded the tolerable limit, with an average value above 1.5 × 103 CFU/g. Attention to food handler training and necessary organizational changes is essential to ensure safe food and promote healthy student eating habits, highlighting the importance of strengthening basic hygiene practices and following the parameters for sanitizing vegetables.

Hygiene doi: 10.3390/hygiene4040039

Authors: Mikiko Tokiya Akiko Matsumoto Yutaro Furukawa Takaomi Kobayashi Takashi Nakano Yoshio Hirota Megumi Hara

Parental willingness is a prerequisite for childhood immunization. This study evaluated whether parental vaccination readiness and willingness, measured using the modified Japanese 7C Vaccination Readiness Scale, predicted vaccination behavior. In December 2021, 2 months before the approval of COVID-19 vaccines for children aged 5–11 years in Japan, 1190 parents of children in this age group were surveyed regarding their intention to vaccinate their children against COVID-19. The survey utilized an online version of the modified 7C Vaccination Readiness Scale, comprising seven components: Confidence, Complacency, Constraints, Calculation, Collective Responsibility, Compliance, and Conspiracy. The modified 7C components were analyzed for model fit and their association with parental intention using structural equation modeling. The scale demonstrated good model fit (goodness-of-fit index: 0.98), with high standardized factor loadings for parental readiness (factor loadings: 0.84). In January 2024, the same parents were surveyed about their children’s COVID-19 vaccination status (valid responses: 536). The relationship between the components of the modified 7C scale and actual vaccination behavior was examined. Parents with low readiness to vaccinate their children against COVID-19 exhibited a significantly higher non-vaccination rate. These findings suggest that the modified 7C scale could support the development of targeted interventions to improve childhood vaccination uptake.

Hygiene doi: 10.3390/hygiene4040038

Authors: Christian Kühnel Falk Gühne

There were some parts of the original publication [...]

Hygiene doi: 10.3390/hygiene4040037

Authors: Emily C. Montgomery Madelyn C. Wicker Tibirni Yusuf Elizabeth Matlock-Buchanan Tomoko Fujiwara Joel D. Bumgardner J. Amber Jennings

Infections are a common post-operative ailment for patients who have received a dental implant or device and can be attributed to biofilm formation in tissue or on the implant. Many current solutions for oral hygiene have side effects and affect the natural oral microbiome. 2-heptylcyclopropane-1-carboxylic acid (2CP) is a medium-chain fatty acid and synthetic diffusible signaling factor that can prevent and disperse biofilm. The purpose of this work was to evaluate an immersion strategy for coating hydroxyapatite (HAp) with 2CP to prevent biofilm attachment on and around natural teeth and dental implants. The release profile of 2CP-loaded and 2CP+oral rinse-loaded HAp coupons (n = 6) was assessed by a 3-day exposure to phosphate buffered saline (PBS). Antimicrobial properties against Porphyromonas gingivalis and cytocompatibility of 2CP-loaded HAp coupons (n = 4) were also assessed alone and in combination with 0.12% chlorhexidine gluconate oral rinse. The majority of 2CP is released by 12 h. 2CP, oral rinse, and 2CP+oral rinse significantly reduced P. gingivalis viability, though direct contact assay demonstrates a significant reduction in Saos-2 viability for oral rinse and 2CP+oral rinse coupons. Immersion or rinsing hydroxyapatite with 2CP could inhibit biofilm-associated dental infections and prevent further complications including caries, gingivitis, and peri-implantitis.

Hygiene doi: 10.3390/hygiene4040036

Authors: Olalekan Chris Akinsulie Victor Ayodele Aliyu Ibrahim Idris Samuel Ajulo Oluwagbemisola Olukogbe Chizaram Ukauwa Jesuferanmi Mary Akinsulie

Infectious diseases are largely preventable, yet they continue to pose a significant threat to public health, particularly among vulnerable populations in developing countries. Basic hygiene practices, especially hand and skin hygiene, have been shown to significantly reduce the risk of the cross-transmission of infections, including those caused by multi-drug-resistant organisms. In light of the growing global concern about antimicrobial resistance, there is an urgent need to review and reinforce these practices. This study provides a general overview of the role that hand hygiene practices play in decreasing infectious diseases by conducting a comprehensive review. Multiple online databases, including Google Scholar, Scopus, and Web of Science, were searched using relevant keywords such as “hygiene practices”, “infectious diseases”, “public health”, “Africa”, and “sanitation”. After filtering the search results for relevancy, selected studies were narratively synthesized to present the latest data on hand hygiene and its impact on infectious diseases. Strengthening hand and skin hygiene, along with environmental sanitation and preventive measures, can help reduce the spread of nosocomial infections. By emphasizing the importance of these fundamental hygiene practices, particularly in regions where the burden of infectious diseases is highest, the development of antibiotic-resistant diseases can be prevented, improving patient safety, and enhancing public health outcomes. Adopting comprehensive hygiene policies, including regular handwashing, is crucial for reducing the prevalence of infectious diseases and improving health outcomes in developing countries.

Hygiene doi: 10.3390/hygiene4040035

Authors: Yuting Han Adenike A. Akinsemolu Christian K. Anumudu Taghi Miri Helen Onyeaka

The symptoms of foodborne illness caused by Bacillus cereus often go unreported, complicating the effectiveness of conventional chemical and physical methods used to inhibit its growth in food production. This challenge, combined with the increasing use of lactic acid bacteria (LAB) in the food industry and consumer preference for minimally processed products, prompted this study. The antibacterial activity of diffusible substances produced by Lactococcus lactis ATCC 11454 against Bacillus cereus NC11143 and Escherichia coli K-12 MG1655 was investigated using a non-contact co-culture model utilising deMann Rogosa and Sharpe broth, with glucose as a carbon source. This study employed plate counting and flow cytometry to assess the impact of these substances on bacterial growth and to analyse their composition and antimicrobial efficacy. The co-culture of Lactococcus lactis ATCC 11454 resulted in the production of a stable antimicrobial peptide, which was heat resistant and acid tolerant. Purification was achieved via ammonium sulphate precipitation and preparative HPLC, yielding a peptide with a molecular mass of 3.3 kDa, with daughter ion fractions similar to nisin A. Antimicrobial activity studies demonstrated that the diffusible substances effectively inhibited B. cereus growth over a period of eight days and exhibited bactericidal activity, killing 99% of the B. cereus cells. Additionally, these substances also inhibited Escherichia coli K-12 MG1655 grown under similar conditions. Comparative analysis revealed that in the co-culture assay, L. lactis produced a 50% higher yield of the antimicrobial peptides compared to pure cultures. Similarly, the specific growth rate of L. lactis was four times higher. With respect to protein purification and concentration, ammonium sulphate precipitation coupled with solid phase extraction was most effective in the purification and concentration of the diffusible substances. The findings provide a basis for utilising bacteriocin-producing strains as a preservation method, offering an alternative to traditional chemical and physical control approaches especially for the food industry.

Hygiene doi: 10.3390/hygiene4040034

Authors: Niki Tertipi Eleni Sfyri Vasiliki Kefala Efstathios Rallis

Background/Objectives: Fungal infections, commonly referred to as mycoses, are widespread and affect various parts of the body. Common types of fungal skin infections include tinea pedis, tinea unguim, tinea cruris, tinea versicolor, and tinea manuum. Athletes, in particular, are at a higher risk of contracting these infections due to several factors inherent to their training and competitive environments. Methods: This research examined the incidence, anatomical distribution, and seasonal occurrence of fungal skin infections among beach volleyball athletes in Greece. The study cohort consisted of 785 participants, with an average age of 28.4 years. Variables recorded included gender, the seasonal periods during which athletes might be more prone to injuries, and the specific body areas affected by fungal infections. In addition, training-related data such as weekly training frequency, and daily training duration were collected. Results: Incidence rates correlated in relation to gender: male, tinea pedis (p < 0.000), tinea manuum (p < 0.010); number of weekly trainings: tinea pedis (p < 0.000), tinea unguium (p < 0.043), tinea cruris (p < 0.008), tinea versicolor (p < 0.038), tinea manuum (p < 0.043); average hours of daily training: tinea unguium (p < 0.039), tinea manuum (p < 0.039). Conclusions: Athletic activities often involve exposure to warm, moist conditions that promote fungal growth. Locker rooms, showers, and equipment can harbor fungi, spreading infections among athletes. Understanding these infections is essential for effective prevention and management, as they can impact performance and health if untreated. Proper hygiene, antifungal treatments, and public health measures are crucial to reducing the risk in athletic settings.

Hygiene doi: 10.3390/hygiene4040033

Authors: Malehlohonolo Ntaote Londele Tyeshani Olanrewaju Oladimeji

Background: Hands are critical vectors for microorganisms that cause healthcare-associated infections (HAIs). Hand hygiene, being done the right way, at the right time, and being given the right priority in a healthcare setting is an effective strategy to reduce HAIs and associated costs. Different strategies have been put in place to help improve hand hygiene compliance among healthcare workers; amongst them, continuous training is advised at all levels of care. Aim: This study, focusing on nursing students, aimed to describe their knowledge, attitudes, and perceptions towards hand hygiene, highlighting their crucial role in maintaining health and preventing infections. Methods: This study was a quantitative cross-sectional study. One hundred and fourteen questionnaires were distributed; 103 were filled out and returned by willing students at Maluti Adventist College, thus a 90% response rate. Results: 62% (64) demonstrated moderate knowledge, all had positive attitudes, and 61% (63) had a positive perception towards hand hygiene. There was a significant (p-value = 0.012) association between knowledge and training. Conclusions and recommendations: These provide valuable input for enhancing the World Health Organization’s (WHO) hand hygiene multimodal strategy, the SAFE LIFE Clean YOUR HAND campaign, and healthcare workers’ teaching curriculum. All stakeholders should implement strategies that prioritize knowledge dissemination to promote effective hand hygiene practices among healthcare workers worldwide.

Hygiene doi: 10.3390/hygiene4040032

Authors: Marta Garzón Prados Laura Jiménez Sampelayo María Isabel Tovar-Gálvez Juan Carlos Sánchez-García Jonathan Cortés-Martín Beatriz Piqueras-Sola Raquel Rodríguez-Blanque

Tuberculosis (TB) is a dangerous disease caused by the bacterium Mycobacterium tuberculosis. Its transmission occurs through aerosols, making healthcare workers particularly vulnerable to it. At the San Cecilio Clinical University Hospital in Granada (HUCSC), preventive measures such as the use of personal protective equipment (PPE) and IGRA (interferon-gamma release assay) tests were implemented to detect latent tuberculosis infection. At HUCSC, between 2022 and 2024, 4.75% (92/1936) of workers were found to have positive IGRA test results, with no significant differences observed between professional categories. However, variability in positive IGRA rates was noted across different medical departments, with pulmonology and sample handling being the most affected. Additionally, a relationship was found between age and the likelihood of obtaining a positive IGRA test result. These findings underscore the importance of targeted preventive measures and a multifaceted approach to controlling and preventing tuberculosis in workplace environments.

Hygiene doi: 10.3390/hygiene4040031

Authors: Maria Antoniadou Maria Chanioti Asteropi Pantelaki Antonios Parasyris Evangelia Piperi Christos Rahiotis

Dental students are frequently exposed to percutaneous injuries (PCIs) due to the nature of their clinical work, which involves sharp instruments and close patient contact. The COVID-19 pandemic further emphasized the need for stringent biosafety measures and the use of personal protective equipment (PPE). Despite these precautions, injuries remain prevalent, highlighting the need for comprehensive education and training in biosafety and infection control. This study investigates the incidence and causes of injuries among undergraduate dental students during clinical sessions. This study was conducted at the Department of Dentistry, National and Kapodistrian University of Athens, focusing on injuries reported from 2021 to 2024. Data were collected through self-reported clinical records. The primary variables assessed included the type of injury, the instrument involved, the clinical procedure performed, and the immediate actions taken post-injury. Serological testing was conducted on students and patients to assess the risk of the transmission of bloodborne pathogens. The findings reveal a high prevalence of injuries, with needles being the most common cause (51.4%), followed by other tools such as dental probes (25.7%) and burs (8.6%). The most frequent injury type is piercing (74.2%), primarily affecting the fingers (88.6%). Periodontal treatments, restorative procedures, and endodontic treatments are the main activities leading to injuries, with 17.1% of injuries being caused by each. No statistically significant results are recorded. Despite regular medical records for most patients treated by injured students, serological testing shows significant positivity rates for HCV and HBV. Notably, most injured students demonstrate their commitment to safety by adhering to recommended post-exposure protocols, including wound cleaning, disinfecting, and serological testing. Furthermore, the impact of COVID-19 heightened the importance of personal protective equipment (PPE) and reinforced occupational health standards. Our study highlights the critical need for enhanced biosafety awareness and training among undergraduate dental students to reduce injury risks.

Hygiene doi: 10.3390/hygiene4030030

Authors: Jamie M. Reedy Theekshana Fernando Silas O. Awuor Eric Omori Omwenga Tatiana Koutchma Richard M. Mariita

Emerging antimicrobial-resistant (AMR) Candida auris presents a formidable global health challenge, causing severe healthcare-associated infections (HAIs) with high mortality rates. Its ability to colonize surfaces and resist standard disinfectants undermines traditional hygiene practices, prompting an urgent need for new strategies. Ultraviolet C (UVC) light offers a promising approach with rapid and broad-spectrum germicidal efficacy. This review examines current literature on UVC LED technology in combating C. auris, highlighting its effectiveness, limitations, and applications in healthcare hygiene. UVC light has potent activity against C. auris, with up to 99.9999% inactivation depending on certain conditions such as microbial load, type of organism, surface, environmental, equipment, and UVC radiation factors. UVC LEDs can effectively combat C. auris, driving down healthcare costs and reducing attributable global mortality. Here, we explore implementation strategies for the targeted disinfection of high-risk areas and equipment, air handling units (AHUs), and water treatment systems. Challenges associated with UVC LED disinfection devices in healthcare settings, current performance limitations, and radiation safety are discussed. This will help in optimizing application protocols for effective disinfection and radiation safety. To further strengthen healthcare facility hygiene practices and curb the global spread of C. auris, recommendations for integrating UVC LED disinfection into infection control programs are shared.

Hygiene doi: 10.3390/hygiene4030029

Authors: Prem K. Sreenivasan Bhojraj Nandlal Pratibha Shashikumar Avinash Bettahalli Shivamallu

Objective: This clinical investigation evaluated oral health improvements after the use of a novel zinc toothpaste in comparison to a control after 3 weeks and 6 weeks of use. Clinical assessments of gingivitis, bleeding and dental plaque evaluated oral health. Methods: Healthy adults meeting the study criteria (plaque index scores ≥ 1.5 and gingival index scores ≥ 1.0) were randomized into a treatment group with 74 participants assigned to the test and 73 participants assigned to the control groups. With the assigned toothpaste, subjects were instructed to brush at home twice a day for one minute. Subjects were evaluated clinically for gingivitis, plaque and bleeding at the baseline, employing 3-week and 6-week examinations. Results: All one hundred and forty-seven (147) subjects completed the 6-week study with no significant differences between groups at baseline (p > 0.05). The test demonstrated progressive improvements at the 3-week and 6-week evaluations as compared to the control (p < 0.001). In comparison to the control, the test demonstrated reductions of 13.4%, 17.0% and 55.3% for gingivitis, dental plaque and gingival bleeding, respectively, at the final visit (p < 0.001). Conclusions: Clinical improvements in gingivitis, dental plaque and gingival bleeding were registered in the group assigned the test toothpaste as compared to the control. The test toothpaste demonstrated statistically significant and progressively improving gingivitis, dental plaque and gingival bleeding scores that were all statistically significant versus the control at each post-treatment assessment.

Hygiene doi: 10.3390/hygiene4030028

Authors: Vitowe Batch Martina Kress Ezekiel Luhanga Maurice Monjerezi Daniel Pfaff Horace Phiri Aggrey Pemba Gama Kelvin Chomanika Anja Schmidt Bianca Duelken Ivana Jurisic Angstone Thembachako Mlangeni Kingsley Mikwamba Gabriella Chiutsi-Phiri Ponyadira Leah Corner Save Kumwenda Chikumbusko Kaonga Susanne Schwan Mphatso Kamndaya Limbikani Matumba

This study assessed knowledge, attitudes, and practices related to food hygiene among mothers from Malawi’s rural communities against the WHO Five Keys to Safer Food (WHO-FKSF) and good pre-and post-harvest practices (GPPHPs) as reference points. Five hundred twenty-two mothers from six rural communities across two districts were selected for the survey. The results indicated limited knowledge among participating mothers regarding managing food hazards, including mycotoxins, bacteria, viruses, and parasites (BVPs). A significant proportion (89.5%) of women reported inconsistent or no handwashing with soap after using the toilet. In addition, 48.7% failed to plant with the first good rains, 38.7% neglected to check for moldy cobs during harvesting, 57.4% dried maize on bare soil, and 99.2% bought maize with noticeable mold. Higher education, knowledge, and positive attitudes were associated with enhanced BVP control practices, while larger households and positive attitudes were linked to improved mold/mycotoxin management (p < 0.05). Mothers showed lower (p < 0.05) knowledge and attitude levels regarding molds than BVPs but demonstrated relatively better practices for mold control. A comprehensive education program based on the WHO Five Keys to Safer Foods, tailored to local socio-cultural norms and incorporating mold and mycotoxin management guidelines, is recommended.

Hygiene doi: 10.3390/hygiene4030027

Authors: Willy Barinem Vidona Charity Willy-Vidona Lekpa Kingdom David Bassey Monday Abia John Nwolim Paul Ukpai Agwu Eze

Background: Over the past few decades, epidemic outbreaks and disease occurrences have become more frequent and widespread in Africa, posing challenges for poor countries in the region and impacting vulnerable populations. Limited resources, inadequate healthcare infrastructure, socioeconomic inequalities, malnutrition, mass gatherings, and lack of proper sanitation and hygiene contribute to Africa’s high burden of epidemics and infectious diseases such as cholera, measles, monkeypox, Ebola viral disease, and COVID-19. This paper reviewed epidemic outbreaks and diseases transmitted through intimate contact in Africa and their impacts on vulnerable populations. Methods: A narrative review approach was adopted to gather and analyze the relevant literature on epidemic outbreaks in African sub-regions. The review encompassed causes, transmission patterns, demographic impact, community susceptibility, preventive measures, and psychosocial risk-taking behaviors. Results: This study emphasizes the importance of addressing the causes of response lapses and the resulting human, material, and economic losses in the region. Effective preventive measures include disease surveillance, early mitigation strategies, contact tracing, personal protective measures, improved political and public health leadership, and socioeconomic equity. However, the review highlights challenges in implementing these measures effectively due to limited resources, delayed detection, and difficulties in scaling up response solutions. Conclusions: There is a need for a comprehensive approach involving health departments, infrastructure development, and addressing socioeconomic factors.

Hygiene doi: 10.3390/hygiene4030026

Authors: Paulina A. Dzianach Gary A. Dykes Norval J. C. Strachan Ken J. Forbes Francisco J. Pérez-Reche

Campylobacter jejuni plays a significant role in human health, food production, and veterinary practice. Biofilm formation is a likely mechanism explaining the survival of C. jejuni in seemingly unfavourable environments, but the underlying mechanisms are poorly understood. We propose a mathematical model to unify various observations regarding C. jejuni biofilm formation. Specifically, we present a cellular automaton with stochastic dynamics that describes both the probability of biofilm initiation and its subsequent growth. Our model incorporates fundamental processes such as cell rearrangement, diffusion of chemical compounds, accumulation of extracellular material, cell growth, lysis, and deactivation due to nutrient scarcity. The model predicts an optimal nutrient concentration that enhances population survival, revealing a trade-off where higher nutrient levels may harm individual cells but benefit the overall population. Our results suggest that the lower biofilm accumulation observed experimentally in aerobic conditions compared to microaerobic conditions may be due to a reduced surface invasion probability of individual cells. However, cells that do manage to invade can generate microcolonies of a similar size under both aerobic and microaerobic conditions. These findings provide new insights into the survival probability and size of C. jejuni biofilms, suggesting potential targets for controlling its biofilm formation in various environments.