{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,22]],"date-time":"2026-06-22T11:47:01Z","timestamp":1782128821204,"version":"3.54.5"},"reference-count":41,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2026,6,22]],"date-time":"2026-06-22T00:00:00Z","timestamp":1782086400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Fondi Ricerca di Ateneo"},{"name":"Fondi Aziendali ASL Caserta"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Microorganisms"],"abstract":"<jats:p>Healthcare-associated infections (HAIs) remain a major public health concern, contributing to increased morbidity, mortality, and antimicrobial resistance. Healthcare workers (HCWs) are recognized as key vehicles in the transmission of nosocomial pathogens, primarily via contaminated hands and medical devices. This study assessed the effectiveness of hand hygiene protocols among HCWs, their correlation with bloodstream infections, and the potential of natural antimicrobial agents as complementary preventive measures. Between January and June 2025, 128 hand samples were collected from HCWs in surgical, intensive care, and internal medicine units of hospitals managed by ASL Caserta (Marcianise, n = 65; Piedimonte Matese, n = 30; Sessa Aurunca, n = 18; Maddaloni, n = 15). Sampling was performed upon entry to clinical areas and after antiseptic handwashing, using Rodac TSA plates to quantify microbial load (CFU\/cm2). Isolates were identified via MALDI-TOF, and multidrug resistance was confirmed using the Phoenix BD system. Microbial growth was detected in 54.7% of samples. Coagulase-negative staphylococci, mainly Staphylococcus epidermidis and S. hominis, accounted for 67.1% of positive cultures, followed by Enterobacteriaceae (28.6%). Comparison with concurrently collected blood cultures revealed potential overlapping pathogens, with Staphylococcus spp. prevalence ranging from 35 to 56% and Gram-negatives from 18 to 39. Selected isolates were further tested for susceptibility to natural antimicrobial agents, derived from hop, red vine leaf, green tea, and pomegranate fruit, as well as thyme essential oil. Thyme essential oil (Thymus vulgaris) demonstrated notable antimicrobial activity, in several cases surpassing that of standard hygiene agents. These findings highlight not only that maintaining high standards of hand hygiene, proper care of invasive devices, and continuous microbiological surveillance is critical for preventing HAIs, but also that incorporating natural antimicrobial compounds into hygiene protocols may provide an effective and sustainable adjunct to reduce microbial contamination and combat infections caused by multidrug-resistant organisms.<\/jats:p>","DOI":"10.3390\/microorganisms14061382","type":"journal-article","created":{"date-parts":[[2026,6,22]],"date-time":"2026-06-22T09:49:55Z","timestamp":1782121795000},"page":"1382","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Monitoring Hygiene Protocols and Exploring Alternatives to Counteract Resistant Pathogens: A Case Study from Southern Italy on Healthcare-Associated Infection Control"],"prefix":"10.3390","volume":"14","author":[{"given":"Enza","family":"Mallardo","sequence":"first","affiliation":[{"name":"Department of Diagnostic Services, Unit of Clinical Pathology, Marcianise Hospital, ASL Caserta, Viale Sossietta Scialla, 81025 Marcianise, Caserta, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Claudio Attilio","family":"Baliano","sequence":"additional","affiliation":[{"name":"Department of Diagnostic Services, Unit of Clinical Pathology, Marcianise Hospital, ASL Caserta, Viale Sossietta Scialla, 81025 Marcianise, Caserta, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Valeria","family":"Pedata","sequence":"additional","affiliation":[{"name":"Department of Diagnostic Services, Unit of Clinical Pathology, Marcianise Hospital, ASL Caserta, Viale Sossietta Scialla, 81025 Marcianise, Caserta, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rosita","family":"Zinzi","sequence":"additional","affiliation":[{"name":"Department of Diagnostic Services, Unit of Clinical Pathology, Marcianise Hospital, ASL Caserta, Viale Sossietta Scialla, 81025 Marcianise, Caserta, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Federica","family":"Mayella","sequence":"additional","affiliation":[{"name":"Department of Diagnostic Services, Unit of Clinical Pathology, Marcianise Hospital, ASL Caserta, Viale Sossietta Scialla, 81025 Marcianise, Caserta, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mauro","family":"Murano","sequence":"additional","affiliation":[{"name":"Department of Diagnostic Services, Unit of Clinical Pathology, Marcianise Hospital, ASL Caserta, Viale Sossietta Scialla, 81025 Marcianise, Caserta, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Antonio","family":"Fascione","sequence":"additional","affiliation":[{"name":"Department of Diagnostic Services, Unit of Clinical Pathology, Marcianise Hospital, ASL Caserta, Viale Sossietta Scialla, 81025 Marcianise, Caserta, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-0377-1714","authenticated-orcid":false,"given":"Giuseppina","family":"Forgione","sequence":"additional","affiliation":[{"name":"Department of Science and Technology, University of Sannio, Via F. De Sanctis Snc, 82100 Benevento, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7149-4085","authenticated-orcid":false,"given":"Daniela","family":"Sateriale","sequence":"additional","affiliation":[{"name":"Department of Science and Technology, University of Sannio, Via F. De Sanctis Snc, 82100 Benevento, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3126-5055","authenticated-orcid":false,"given":"Caterina","family":"Pagliarulo","sequence":"additional","affiliation":[{"name":"Department of Science and Technology, University of Sannio, Via F. De Sanctis Snc, 82100 Benevento, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,6,22]]},"reference":[{"key":"ref_1","unstructured":"European Centre for Disease Prevention and Control (2026, May 16). Incidence and Attributable Mortality of Healthcare-Associated Infections in Intensive Care Units in Europe, 2008\u20132012. Available online: https:\/\/www.ecdc.europa.eu\/en\/publications-data\/incidence-and-attributable-mortality-healthcare-associated-infections-intensive."},{"key":"ref_2","unstructured":"World Health Organization (2026, March 12). World Health Statistics 2017: Monitoring Health for the SDGs, Sustainable Development Goals. Available online: https:\/\/www.who.int\/publications\/i\/item\/9789241565486."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"38","DOI":"10.1186\/s13756-022-01075-1","article-title":"Impact of Environmental Hygiene Interventions on Healthcare-Associated Infections and Patient Colonization: A Systematic Review","volume":"11","author":"Peters","year":"2022","journal-title":"Antimicrob. Resist. Infect. Control"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"2510875","DOI":"10.1155\/2019\/2510875","article-title":"Recent Advances in Investigation, Prevention, and Management of Healthcare-Associated Infections (HAIs): Resistant Multidrug Strain Colonization and Its Risk Factors in an Intensive Care Unit of a University Hospital","volume":"2019","author":"Zaha","year":"2019","journal-title":"BioMed Res. Int."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Garvey, M. (2023). Medical Device-Associated Healthcare Infections: Sterilization and the Potential of Novel Biological Approaches to Ensure Patient Safety. Int. J. Mol. Sci., 25.","DOI":"10.3390\/ijms25010201"},{"key":"ref_6","unstructured":"European Centre for Disease Prevention and Control (ECDC) (2026, May 16). Point Prevalence Survey of Healthcare-Associated Infections and Antimicrobial Use in European Acute Care Hospitals\u20142022\u20132023. Available online: https:\/\/www.ecdc.europa.eu\/en\/publications-data\/PPS-HAI-AMR-acute-care-europe-2022-2023."},{"key":"ref_7","unstructured":"EpiCentro Istituto Superiore di Sanit\u00e0 (2026, May 16). Piano Nazionale Di Contrasto All\u2019Antibiotico-Resistenza (PNCAR) 2022\u20132025. Available online: https:\/\/www.epicentro.iss.it\/antibiotico-resistenza\/pncar-2022#writers."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Borkow, G. (2014). The Role of Contaminated Surfaces in the Transmission of Nosocomial Pathogens. Use of Biocidal Surfaces for Reduction of Healthcare Acquired Infections, Springer International Publishing.","DOI":"10.1007\/978-3-319-08057-4"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1307","DOI":"10.1016\/S0140-6736(00)02814-2","article-title":"Effectiveness of a Hospital-Wide Programme to Improve Compliance with Hand Hygiene","volume":"356","author":"Pittet","year":"2000","journal-title":"Lancet"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"819","DOI":"10.1016\/j.amjmed.2010.03.021","article-title":"The Clinical and Prognostic Importance of Positive Blood Cultures in Adults","volume":"123","author":"Pien","year":"2010","journal-title":"Am. J. Med."},{"key":"ref_11","unstructured":"(2004). Microbiology of Food and Animal Feeding Stuffs\u2014Horizontal Methods for Sampling Techniques from Surfaces Using Contact Plates and Swabs. (Standard No. ISO 18593:2004)."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"174","DOI":"10.1016\/j.copbio.2011.08.007","article-title":"Polyphenols as Antimicrobial Agents","volume":"23","author":"Daglia","year":"2012","journal-title":"Curr. Opin. Biotechnol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"4737","DOI":"10.1007\/s13197-014-1533-1","article-title":"Antibacterial Activity and Mode of Action of Selected Glucosinolate Hydrolysis Products against Bacterial Pathogens","volume":"52","author":"Borges","year":"2015","journal-title":"J. Food Sci. Technol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"665","DOI":"10.1128\/CMR.00020-14","article-title":"Controlling Hospital-Acquired Infection: Focus on the Role of the Environment and New Technologies for Decontamination","volume":"27","author":"Dancer","year":"2014","journal-title":"Clin. Microbiol. Rev."},{"key":"ref_15","unstructured":"(2018). Microbiology of the Food Chain\u2014Horizontal Methods for Surface Sampling (Standard No. ISO 18593:2018)."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Sateriale, D., Forgione, G., De Cristofaro, G.A., Continisio, L., Pagliuca, C., Colicchio, R., Salvatore, P., Paolucci, M., and Pagliarulo, C. (2024). Eco-Friendly Sanitization of Indoor Environments: Effectiveness of Thyme Essential Oil in Controlling Bioaerosol Levels and Disinfecting Surfaces. BioTech, 13.","DOI":"10.3390\/biotech13020012"},{"key":"ref_17","unstructured":"Clinical and Laboratory Standards Institute (CLSI) (2022). Performance Standards for Antimicrobial Susceptibility Testing, Clinical and Laboratory Standards Institute (CLSI). [32nd ed.]. CLSI Supplement M100."},{"key":"ref_18","unstructured":"Clinical and Laboratory Standards Institute (CLSI) (2022). Method for Dilution Antimicrobial Susceptibility Tests for Bacteria That Grow Aerobically, Clinical and Laboratory Standards Institute (CLSI)."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"694","DOI":"10.1089\/sur.2014.011","article-title":"Surface Contamination in Operating Rooms: A Risk for Transmission of Pathogens?","volume":"15","author":"Yezli","year":"2014","journal-title":"Surg. Infect."},{"key":"ref_20","unstructured":"World Health Organization (2009). WHO Guidelines on Hand Hygiene in Health Care: First Global Patient Safety Challenge\u2014Clean Care Is Safer Care, World Health Organization."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1177\/1420326X03012001002","article-title":"The Airborne Transmission of Infection in Hospital Buildings: Fact or Fiction?","volume":"12","author":"Beggs","year":"2003","journal-title":"Indoor Built Environ."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"S747","DOI":"10.1098\/rsif.2009.0228.focus","article-title":"Control of Airborne Infectious Diseases in Ventilated Spaces","volume":"6","author":"Nielsen","year":"2009","journal-title":"J. R. Soc. Interface"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"436","DOI":"10.1016\/j.buildenv.2012.09.011","article-title":"Bioaerosol Deposition in Single and Two-Bed Hospital Rooms: A Numerical and Experimental Study","volume":"59","author":"King","year":"2013","journal-title":"Build. Environ."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"107373","DOI":"10.1016\/j.buildenv.2020.107373","article-title":"Full-Scale Experimental and Numerical Study of Bioaerosol Characteristics against Cross-Infection in a Two-Bed Hospital Ward","volume":"186","author":"Liu","year":"2020","journal-title":"Build. Environ."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"687","DOI":"10.1086\/660363","article-title":"The Role Played by Contaminated Surfaces in the Transmission of Nosocomial Pathogens","volume":"32","author":"Otter","year":"2011","journal-title":"Infect. Control Hosp. Epidemiol."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Kramer, A., Schwebke, I., and Kampf, G. (2006). How Long Do Nosocomial Pathogens Persist on Inanimate Surfaces? A Systematic Review. BMC Infect. Dis., 6.","DOI":"10.1186\/1471-2334-6-130"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"2","DOI":"10.1111\/j.1600-0668.2006.00445.x","article-title":"Role of Ventilation in Airborne Transmission of Infectious Agents in the Built Environment\u2014A Multidisciplinary Systematic Review","volume":"17","author":"Li","year":"2007","journal-title":"Indoor Air"},{"key":"ref_28","first-page":"1","article-title":"Guidelines for Environmental Infection Control in Health-Care Facilities. Recommendations of CDC and the Healthcare Infection Control Practices Advisory Committee (HICPAC)","volume":"52","author":"Sehulster","year":"2003","journal-title":"Morb. Mortal. Wkly. Rep. Recomm. Rep. RR"},{"key":"ref_29","unstructured":"ANSI, ASHRAE, and ASHE (2026, May 16). ANSI\/ASHRAE\/ASHE Standard 170-2025: Ventilation of Health Care Facilities. Available online: https:\/\/www.ashe.org\/standard170-2025."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1765","DOI":"10.2147\/RMHP.S269315","article-title":"Strategies to Prevent Healthcare-Associated Infections: A Narrative Overview","volume":"13","author":"Haque","year":"2020","journal-title":"Risk Manag. Healthc. Policy"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"880","DOI":"10.1017\/ice.2023.280","article-title":"Environmental Cleaning and Disinfection in the Operating Room: A Systematic Scoping Review through a Human Factors and Systems Engineering Lens","volume":"45","author":"Xie","year":"2024","journal-title":"Infect. Control Hosp. Epidemiol."},{"key":"ref_32","unstructured":"European Centre for Disease Prevention and Control (ECDC) (2026, May 16). Healthcare-Associated Infections Acquired in Intensive Care Units\u2014Annual Epidemiological Report for 2021. Available online: https:\/\/www.ecdc.europa.eu\/en\/publications-data\/healthcare-associated-infections-acquired-intensive-care-units-annual."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"62","DOI":"10.1016\/j.enceco.2023.01.003","article-title":"Occurrence, Effects, and Ecological Risks of Chemicals in Sanitizers and Disinfectants: A Review","volume":"5","author":"Musee","year":"2023","journal-title":"Environ. Chem. Ecotoxicol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"36","DOI":"10.1007\/s11274-025-04775-z","article-title":"A Review on Novel Strategies to Combat Multidrug Resistance in Pathogenic Bacteria Exploiting Synergism between Essential Oil and Antibiotics","volume":"42","author":"Mazumder","year":"2026","journal-title":"World J. Microbiol. Biotechnol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"110354","DOI":"10.1016\/j.lwt.2020.110354","article-title":"Thymol as a Critical Component of Thymus Vulgaris L. Essential Oil Combats Pseudomonas Aeruginosa by Intercalating DNA and Inactivating Biofilm","volume":"136","author":"Liu","year":"2021","journal-title":"LWT"},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Iseppi, R., Mariani, M., Cond\u00f2, C., Sabia, C., and Messi, P. (2021). Essential Oils: A Natural Weapon against Antibiotic-Resistant Bacteria Responsible for Nosocomial Infections. Antibiotics, 10.","DOI":"10.3390\/antibiotics10040417"},{"key":"ref_37","first-page":"92","article-title":"Antibiofilm Activity of Natural Products: Promising Strategies for Combating Microbial Biofilms","volume":"4","year":"2020","journal-title":"Ann. Public Health Rep."},{"key":"ref_38","unstructured":"U.S. Food and Drug Administration (2026, May 16). Generally Recognized as Safe (GRAS), Available online: https:\/\/www.fda.gov\/food\/food-ingredients-packaging\/generally-recognized-safe-gras."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1451","DOI":"10.3390\/ph6121451","article-title":"Effect of Essential Oils on Pathogenic Bacteria","volume":"6","author":"Nazzaro","year":"2013","journal-title":"Pharmaceuticals"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"446","DOI":"10.1016\/j.fct.2007.09.106","article-title":"Biological Effects of Essential Oils\u2014A Review","volume":"46","author":"Bakkali","year":"2008","journal-title":"Food Chem. Toxicol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"3012462","DOI":"10.1155\/2016\/3012462","article-title":"Antimicrobial Properties of Plant Essential Oils against Human Pathogens and Their Mode of Action: An Updated Review","volume":"2016","author":"Swamy","year":"2016","journal-title":"Evid.-Based Complement. Altern. Med."}],"container-title":["Microorganisms"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2607\/14\/6\/1382\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,6,22]],"date-time":"2026-06-22T10:48:08Z","timestamp":1782125288000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2607\/14\/6\/1382"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,6,22]]},"references-count":41,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2026,6]]}},"alternative-id":["microorganisms14061382"],"URL":"https:\/\/doi.org\/10.3390\/microorganisms14061382","relation":{},"ISSN":["2076-2607"],"issn-type":[{"value":"2076-2607","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,6,22]]}}}