Name: a method to test the efficacy of handwashing for the removal of emerging infectious pathogens
Uploaded: 2017-06-07T14:00:00.000+00:00
Duration: 9 min 2 s
Description: Watch this Scientific Journal Video about A Method to Test the Efficacy of Handwashing for the Removal of Emerging Infectious Pathogens at JoVE.com

Questo lavoro è stato sostenuto dall&#39;Agenzia degli Stati Uniti per lo Sviluppo Internazionale, Ufficio di Assistenza Estera Del Disastro (AID-OFDA-A-15-00026). Marlene Wolfe è stata sostenuta dalla National Science Foundation (sovvenzione 0966093).

<ol>
	<li>Kampf, G., Kramer, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Epidemiologic+Background+of+Hand+Hygiene+and+Evaluation+of+the+Most+Important+Agents+for+Scrubs+and+Rubs.">Epidemiologic Background of Hand Hygiene and Evaluation of the Most Important Agents for Scrubs and Rubs.</a> Clin Microbiol Rev. 17 (4), 863-893 (2004).</li><li>Miller, T., Patrick, D., Ormrod, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hand+decontamination:+influence+of+common+variables+on+hand-washing+efficiency.">Hand decontamination: influence of common variables on hand-washing efficiency.</a> Healthc Infect. 16 (1), 18 (2013).</li><li>Jensen, D. A., Danyluk, M. D., Harris, L. J., Schaffner, D. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantifying+the+effect+of+hand+wash+duration,+soap+use,+ground+beef+debris,+and+drying+methods+on+the+removal+of+Enterobacter+aerogenes+on+hands.">Quantifying the effect of hand wash duration, soap use, ground beef debris, and drying methods on the removal of Enterobacter aerogenes on hands.</a> J Food Prot. 78 (4), 685-690 (2015).</li><li>Girou, E., Loyeau, S., Legrand, P., Oppein, F., Brun-Buisson, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Efficacy+of+handrubbing+with+alcohol+based+solution+versus+standard+handwashing+with+antiseptic+soap:+randomised+clinical+trial.">Efficacy of handrubbing with alcohol based solution versus standard handwashing with antiseptic soap: randomised clinical trial.</a> BMJ. 325 (7360), 362 (2002).</li><li>Kac, G., Podglajen, I., Gueneret, M., Vaupr&#233;, S., Bissery, A., Meyer, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microbiological+evaluation+of+two+hand+hygiene+procedures+achieved+by+healthcare+workers+during+routine+patient+care:+a+randomized+study.">Microbiological evaluation of two hand hygiene procedures achieved by healthcare workers during routine patient care: a randomized study.</a> J Hosp Infect. 60 (1), 32-39 (2005).</li><li>Lages, S. L. S., Ramakrishnan, M. A., Goyal, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In-vivo+efficacy+of+hand+sanitisers+against+feline+calicivirus:+a+surrogate+for+norovirus.">In-vivo efficacy of hand sanitisers against feline calicivirus: a surrogate for norovirus.</a> J Hosp Infect. 68 (2), 159-163 (2008).</li><li>Holton, R. H., Huber, M. A., Terezhalmy, G. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antimicrobial+efficacy+of+soap+and+water+hand+washing+versus+an+alcohol-based+hand+cleanser.">Antimicrobial efficacy of soap and water hand washing versus an alcohol-based hand cleanser.</a> Tex Dent J. 126 (12), 1175-1180 (2009).</li><li>Salmon, S., Truong, A. T., Nguyen, V. H., Pittet, D., McLaws, M. -. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Health+care+workers'+hand+contamination+levels+and+antibacterial+efficacy+of+different+hand+hygiene+methods+used+in+a+Vietnamese+hospital.">Health care workers' hand contamination levels and antibacterial efficacy of different hand hygiene methods used in a Vietnamese hospital.</a> Am J Infect Control. 42 (2), 178-181 (2014).</li><li>Steinmann, J., Nehrkorn, R., Meyer, A., Becker, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Two+in-vivo+protocols+for+testing+virucidal+efficacy+of+handwashing+and+hand+disinfection.">Two in-vivo protocols for testing virucidal efficacy of handwashing and hand disinfection.</a> Int J Hyg Environ Health. 196 (5), 425-436 (1995).</li><li>Weber, D. J., Sickbert-Bennett, E., Gergen, M. F., Rutala, W. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Efficacy+of+selected+hand+hygiene+agents+used+to+remove+Bacillus+atrophaeus+(a+surrogate+of+Bacillus+anthracis)+from+contaminated+hands.">Efficacy of selected hand hygiene agents used to remove Bacillus atrophaeus (a surrogate of Bacillus anthracis) from contaminated hands.</a> JAMA. 289 (10), 1274-1277 (2003).</li><li>Grayson, M. L., Melvani, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Efficacy+of+Soap+and+Water+and+Alcohol-Based+Hand-Rub+Preparations+against+Live+H1N1+Influenza+Virus+on+the+Hands+of+Human+Volunteers.">Efficacy of Soap and Water and Alcohol-Based Hand-Rub Preparations against Live H1N1 Influenza Virus on the Hands of Human Volunteers.</a> Clin Infect Dis. 48 (3), 285-291 (2009).</li><li>Oughton, M. T., Loo, V. G., Dendukuri, N., Fenn, S., Libman, M. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hand+hygiene+with+soap+and+water+is+superior+to+alcohol+rub+and+antiseptic+wipes+for+removal+of+Clostridium+difficile.">Hand hygiene with soap and water is superior to alcohol rub and antiseptic wipes for removal of Clostridium difficile.</a> Infect Control Hosp Epidemiol. 30 (10), 939-944 (2009).</li><li>Liu, P., Yuen, Y., Hsiao, H. -. M., Jaykus, L. -. A., Moe, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effectiveness+of+liquid+soap+and+hand+sanitizer+against+Norwalk+virus+on+contaminated+hands.">Effectiveness of liquid soap and hand sanitizer against Norwalk virus on contaminated hands.</a> Appl Environ Micro. 76 (2), 394-399 (2010).</li><li>Savolainen-Kopra, C., Korpela, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Single+treatment+with+ethanol+hand+rub+is+ineffective+against+human+rhinovirus--hand+washing+with+soap+and+water+removes+the+virus+efficiently.">Single treatment with ethanol hand rub is ineffective against human rhinovirus--hand washing with soap and water removes the virus efficiently.</a> J Med Virol. 84 (3), 543-547 (2012).</li><li>Tuladhar, E., Hazeleger, W. C., Koopmans, M., Zwietering, M. H., Duizer, E., Beumer, R. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reducing+viral+contamination+from+finger+pads:+handwashing+is+more+effective+than+alcohol-based+hand+disinfectants.">Reducing viral contamination from finger pads: handwashing is more effective than alcohol-based hand disinfectants.</a> J Hosp Infect. 90 (3), 226-234 (2015).</li><li>Steinmann, J., Paulmann, D., Becker, B., Bischoff, B., Steinmann, E., Steinmann, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+virucidal+activity+of+alcohol-based+hand+sanitizers+versus+antimicrobial+hand+soaps+in+vitro+and+in+vivo.">Comparison of virucidal activity of alcohol-based hand sanitizers versus antimicrobial hand soaps in vitro and in vivo.</a> J Hosp Infect. 82 (4), 277-280 (2012).</li><li>de Aceituno, A. F., Bartz, F. E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ability+of+Hand+Hygiene+Interventions+Using+Alcohol-Based+Hand+Sanitizers+and+Soap+To+Reduce+Microbial+Load+on+Farmworker+Hands+Soiled+during+Harvest.">Ability of Hand Hygiene Interventions Using Alcohol-Based Hand Sanitizers and Soap To Reduce Microbial Load on Farmworker Hands Soiled during Harvest.</a> J Food Protect. 78 (11), 2024-2032 (2015).</li><li>Boyce, J. M., Pittet, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Guideline+for+Hand+Hygiene+in+Health-Care+Settings+Recommendations+of+the+Healthcare+Infection+Control+Practices+Advisory+Committee+and+the+HICPAC/SHEA/APIC/IDSA+Hand+Hygiene+Task+Force.">Guideline for Hand Hygiene in Health-Care Settings Recommendations of the Healthcare Infection Control Practices Advisory Committee and the HICPAC/SHEA/APIC/IDSA Hand Hygiene Task Force.</a> Infect Control Hosp Epidemiol. 23 (12 Suppl), S3-S40 (2002).</li><li>. UNDP Medical Waste Experts Assessment and Recommendations Regarding Management of Ebola-Contaminated Waste Available from: <a target="_blank" href="https://noharm-global.org/sites/default/files/documents-files/3127/Report%20to%20WHO%20WASH%20and%20Geneva%20on%20Ebola%20final.pdf">https://noharm-global.org/sites/default/files/documents-files/3127/Report%20to%20WHO%20WASH%20and%20Geneva%20on%20Ebola%20final.pdf</a> (2015)</li><li>Hopman, J., Kubilay, Z., Allen, T., Edrees, H., Pittet, D., Allegranzi, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Efficacy+of+chlorine+solutions+used+for+hand+hygiene+and+gloves+disinfection+in+Ebola+settings:+a+systematic+review.">Efficacy of chlorine solutions used for hand hygiene and gloves disinfection in Ebola settings: a systematic review.</a> Antimicrob Resist Infect Control. 4 (1), 1 (2015).</li><li>Lowbury, E. J. L., Lilly, H. A., Bull, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Disinfection+of+hands:+removal+of+transient+organisms.">Disinfection of hands: removal of transient organisms.</a> BMJ. 2 (5403), 230-233 (1964).</li><li>Edmonds, S. L., Zapka, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effectiveness+of+Hand+Hygiene+for+Removal+of+Clostridium+difficile+Spores+from+Hands.">Effectiveness of Hand Hygiene for Removal of Clostridium difficile Spores from Hands.</a> Infect Control Hosp Epidemiol. 34 (3), 302-305 (2013).</li><li>Rotter, M. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=150+years+of+hand+disinfection-Semmelweis'+heritage.">150 years of hand disinfection-Semmelweis' heritage.</a> Hyg Med. (22), 332-339 (1997).</li><li>Hitomi, S., Baba, S., Yano, H., Morisawa, Y., Kimura, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antimicrobial+effects+of+electrolytic+products+of+sodium+chloride--comparative+evaluation+with+sodium+hypochlorite+solution+and+efficacy+in+handwashing.">Antimicrobial effects of electrolytic products of sodium chloride--comparative evaluation with sodium hypochlorite solution and efficacy in handwashing.</a> Kansensh&#333;gaku Zasshi. 72 (11), 1176-1181 (1998).</li><li>. Standard E1174-13. Standard Test Method for Evaluation of the Effectiveness of Health Care Personnel Handwash Formulations Available from: <a target="_blank" href="https://www.astm.org/">https://www.astm.org/</a> (2013)</li><li>Casanova, L. M., Weaver, S. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+eluents+for+the+recovery+of+an+enveloped+virus+from+hands+by+whole-hand+sampling.">Evaluation of eluents for the recovery of an enveloped virus from hands by whole-hand sampling.</a> J Appl Microbiol. 118 (5), 1210-1216 (2015).</li><li>Sinclair, R. G., Rose, J. B., Hashsham, S. A., Gerba, C. P., Haas, C. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Criteria+for+Selection+of+Surrogates+Used+To+Study+the+Fate+and+Control+of+Pathogens+in+the+Environment.">Criteria for Selection of Surrogates Used To Study the Fate and Control of Pathogens in the Environment.</a> Appl Environ Microbiol. 78 (6), 1969-1977 (2012).</li><li>Held, E., Skoet, R., Johansen, J. D., Agner, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+hand+eczema+severity+index+(HECSI):+A+scoring+system+for+clinical+assessment+of+hand+eczema.+A+study+of+inter-+and+intraobserver+reliability.">The hand eczema severity index (HECSI): A scoring system for clinical assessment of hand eczema. A study of inter- and intraobserver reliability.</a> Br J Dermatol. 152 (2), 302-307 (2005).</li><li>. Method 1604: Total Coliforms and Escherichia coli in Water by Membrane Filtration Using a Simultaneous Detection Technique (MI Medium) Available from: <a target="_blank" href="https://www.epa.gov/sites/production/files/2015-08/documents/method_1604_2002.pdf">https://www.epa.gov/sites/production/files/2015-08/documents/method_1604_2002.pdf</a> (2002)</li><li>Adams, M. H., Anderson, E. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Bacteriophages. , (1959).</li><li>Kao, L. S., Green, C. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Analysis+of+Variance:+Is+There+a+Difference+in+Means+and+What+Does+It+Mean?.">Analysis of Variance: Is There a Difference in Means and What Does It Mean?.</a> The Journal of surgical research. 144 (1), 158-170 (2008).</li><li>Schutz, R. W., Gessaroli, M. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Analysis+of+Repeated+Measures+Designs+Involving+Multiple+Dependent+Variables.">The Analysis of Repeated Measures Designs Involving Multiple Dependent Variables.</a> Research Quarterly for Exercise and Sport. 58 (2), 132-149 (1987).</li><li>Woolwine, J. D., Gerberding, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+testing+method+on+apparent+activities+of+antiviral+disinfectants+and+antiseptics.">Effect of testing method on apparent activities of antiviral disinfectants and antiseptics.</a> Antimicrob Agents Chemother. 39 (4), 921-923 (1995).</li></ol>

Gli autori dichiarano di non avere interessi finanziari concorrenti.

The method described here provides an approach for testing handwashing efficacy in a controlled laboratory setting. This method highlights the use of human volunteers and surrogate, non-infectious organisms. Using the method, it was possible to demonstrate differences in: 1) the efficacy of handwashing methods and 2) organism persistence in rinse water. The purpose of presenting this protocol is to provide a general framework that can be adapted to test a wide range of surrogate organisms and handwashing methods relevant to infectious disease.
During the use of the method, two key data quality recommendations were noted as important. First, the inoculate must be applied both as similarly as possible across the rounds of testing and in a manner, that minimizes loss. This is to ensure that sufficient inoculate is applied to the hands to allow for statistically significant results. Second, be sure to complete the "cleansing wash" step, in which the protocol is performed without handwashing prior to testing, as previous work has shown that there are likely to be significant differences between a first wash and subsequent washes, but not between subsequent washes performed after a cleansing round29. Additionally, this step clears residual hand contamination, which would interfere with results. 
The main limitation of this protocol is that it can be uncomfortable for volunteers. During each round of testing, which lasted about 2 h, volunteers' hands became cold. Some volunteers reported mild pain from their hands being constantly wet. Additionally, after a few rounds of testing, volunteers' hands became supersaturated, no longer fully drying between rounds. Although the randomization of the order of handwashing methods for each volunteer accounted for supersaturation, it is possible that the supersaturation could act as a confounding or modifying factor in this type of testing. To address this limitation, it is recommended that volunteers are appraised of this risk during consent disclosures and are reminded of their right to drop out of the study at any time. Volunteers should not undergo testing for more than 2 h per day to allow time for the hands to return to a baseline state and to minimize discomfort. A second limitation is the need to use a surrogate organism or non-infectious variant of a pathogenic organism to protect the health of volunteers. This might cause concern about the generalizability of results. However, for some pathogenic organisms (such as the Ebola virus), this limitation cannot be ethically overcome. Care must be taken during surrogate organism selection. Lastly, this is a laboratory study on efficacy. Results may only translate to effective disease prevention in real-word contexts where handwashing methods are made accessible to those in need and are used properly and consistently.
This protocol draws on previous work on handwashing efficacy but attempts to streamline methods and emphasizes the use of human hands (rather than surrogate surfaces) for testing. Additionally, rinse water is a transmission risk that had previously not been assessed. Existing studies on handwashing efficacy vary in methodology, leading to non-comparable data. We hope that standardized protocols for conducting handwashing method comparisons will encourage comparable and replicable results. Previous work has demonstrated that in vitro testing on surrogate surfaces such as pig skin, where, for example, the actual Ebola virus could be used, produced results that do not match those found after testing on human hands38. Therefore, a method using human hands and surrogate or non-infectious organisms is currently the best available approach to estimate handwashing efficacy and rinse water persistence for infectious microorganisms.
Handwashing is critical to prevent disease transmission. However, there is a lack of evidence on the comparative efficacy of handwashing methods that are commonly recommended. This protocol can be used to generate evidence about handwashing efficacy and rinse water persistence. This is especially important for infectious diseases with the potential to cause large outbreaks, such as the Ebola virus. We hope that other researchers will find this protocol useful to generate much-needed additional evidence on handwashing method efficacy and rinse water persistence that will assist in developing recommendations to reduce the transmission of infectious diseases.

La lavata a mano è ampiamente raccomandata per prevenire la diffusione delle malattie, in particolare quelle trasmesse dalla via fecale-orale o aerea, comprese le malattie diarrea e respiratoria 1 . Sorprendentemente, non vi è alcuna prova comparabile sull&#39;efficacia dei metodi di lavaggio a mano, come la lavaggio a mano con sapone e acqua (HWWS) e con il liquido sanitario a base di alcol (ABHS), sulla rimozione di organismi dalle mani. La ricerca iniziale ha scoperto che l&#39;azione meccanica della lavaggio a mano, al contrario del metodo di lavaggio a mano, può rappresentare la maggior parte delle rimozioni dell&#39;organismo 2 , 3 . Inoltre, non vi è alcuna prova comparativa su quale metodo di lavaggio a mano è più efficace. In una revisione informale della letteratura, sono stati identificati 14 studi che hanno confrontato l&#39;efficacia del sapone e del sanitizzatore delle mani sulla rimozione degli organismi. Di questi studi, cinque hanno trovato ABHS per essere più efficaci 4 , </Sup&gt; 5 , 6 , 7 , 8 , sette trovarono HWWS per essere più efficaci 9 , 10 , 11 , 12 , 13 , 14 , 15 e due non trovarono alcuna differenza significativa tra i metodi 16 , 17 . Questi risultati sono incoerenti e non affrontano il rischio continuo di malattia dalla persistenza degli organismi nell&#39;acqua di risciacquo dopo la lavata a mano. Nel complesso, l&#39;evidenza sull&#39;efficacia comparata dei metodi di lavaggio a mano per la rimozione di patogeni causanti malattie infettive è limitata. Questa prova limitata ha portato all&#39;incertezza circa quali metodi sono più appropriati nelle impostazioni dell&#39;epidemia. Per esempio, Durante l&#39;epidemia di virus di Ebola (EVD) in Africa occidentale dal 2013 al 2016, numerosi intervistati internazionali hanno fornito raccomandazioni contraddittorie per HWWS, ABHS o soluzioni di cloro da 0,05%. Médecins Sans Frontières (MSF) raccomanda l&#39;uso di una soluzione di cloro da 0,05% per la lavata a mano, mentre l&#39;Organizzazione Mondiale della Sanità (WHO) raccomanda HWWS o ABHS (se le mani non sono visibilmente sporche). L&#39;OMS va addirittura affermando che il cloro non deve essere utilizzato a meno che non siano disponibili altre opzioni, perché è meno efficace di altri metodi a causa della domanda di cloro esercitata dalla pelle 18 , 19 , 20 , 21 , 22 . Inoltre, le soluzioni di cloro vengono comunemente prodotte da quattro diversi composti di cloro, tra cui l&#39;ipoclorito ad alta test (HTH), l&#39;ipoclorito di sodio localizzato e stabilizzato (NaOCl) e la sodioIodicloroisocianurato (NaDCC). Una revisione sistematica commissionata dall&#39;OMS in risposta all&#39;epidemia di EVD in Africa occidentale ha recentemente trovato solo quattro studi che indagano sull&#39;efficacia comparativa della lavaggio a mano con cloro 23 . Questi studi hanno inoltre prodotto risultati conflittuali e nessuno di questi studi ha utilizzato la concentrazione raccomandata di cloro dello 0,05% per la lavata a mano o studiati microorganismi simili al virus Ebola 10 , 24 , 25 , 26 , 27 . Pertanto, le raccomandazioni non sono state ritenute basate su prove e non era chiaro quali raccomandazioni erano più efficaci. Sono necessarie ulteriori ricerche per confrontare gli approcci di lavaggio a mano per prevenire la diffusione degli agenti patogeni, poiché gli interventi di lavaggio a mano sono uno strumento importante per prevenire la trasmissione di malattie epidemiche. Questi hLe raccomandazioni di lavaggio devono essere basate su prove. Pertanto, è stato sviluppato un metodo per testare l&#39;efficacia della lavaggio e la persistenza dell&#39;acqua di risciacquo, eseguita con surrogati o agenti patogeni non infettivi, 2 , 28 e 29 . I risultati del campione, utilizzando Phi6 come surrogato per il virus Ebola e utilizzando Escherichia coli come un comune indicatore organismo, sono qui presentati. In questo protocollo viene presentata l&#39;efficacia della lavaggio e il risciacquo delle prove di persistenza dell&#39;acqua.

<table><tbody><tr><td>Soap bar</td><td>Dove</td><td>White Beauty Bar soap</td><td></td></tr><tr><td>Alcohol-based hand sanitizer</td><td>Purell</td><td>Advanced Instant Hand Sanitizer with 70% Ethyl Alcohol</td><td></td></tr><tr><td>HTH Powder</td><td>Acros Organics</td><td>300340010</td><td></td></tr><tr><td>NaDCC Powder</td><td>Medentech</td><td>Klorsept granules</td><td></td></tr><tr><td>NaOCl Solution</td><td>Acros Organics</td><td>419550010</td><td></td></tr><tr><td>Electrochlorinator</td><td>AquaChlor</td><td></td><td></td></tr><tr><td>Iodometric titrator</td><td>Hach</td><td>1690001</td><td></td></tr><tr><td>Bovine serum albumin</td><td>MP Biomedicals</td><td>NC0117242</td><td></td></tr><tr><td>Tryptone</td><td>Fisher</td><td>BP1421-100</td><td></td></tr><tr><td>Bovine Mucin</td><td>EMD Milipore</td><td>49-964-3500MG</td><td></td></tr><tr><td>0.22 &amp;micro;m Filter</td><td>EMD Milipore</td><td>GVWP04700</td><td></td></tr><tr><td>NaCl</td><td>Fisher</td><td>BP358-1</td><td></td></tr><tr><td>Skin pH probe</td><td>Hanna Instruments</td><td>H199181</td><td></td></tr><tr><td>Large Whirlpak Sample Bag</td><td>Nasco</td><td>B01447WA</td><td></td></tr><tr><td>Small Whirlpak Sample Bag</td><td>Nasco</td><td>B01323WA</td><td></td></tr><tr><td>Funnel bottle</td><td>Thermo Scientific</td><td>3120850001</td><td>You may drill an appropriately sized hole in the lid of a bottle to form a funnel that will dispense water at the appropriate flow rate</td></tr><tr><td>Ethanol</td><td>ThermoScientific</td><td>615090010</td><td>Mix with water to produce 70% ethanol</td></tr><tr><td>Spray bottle</td><td>Qorpak</td><td>PLC06934</td><td></td></tr><tr><td>&lt;em&gt;E. coli&lt;/em&gt;</td><td>ATCC</td><td>25922</td><td></td></tr><tr><td>LB Broth</td><td>Fisher BioReagents</td><td>BP1426-2</td><td></td></tr><tr><td>LB Agar</td><td>Fisher BioReagents</td><td>BP1425-500</td><td></td></tr><tr><td>Sterile loop</td><td>Globe Scientific</td><td>22-170-204</td><td></td></tr><tr><td>Phi6</td><td>HER</td><td>102</td><td></td></tr><tr><td>Nutrient broth</td><td>BD Difco</td><td>BD 247110</td><td></td></tr><tr><td>GeneQuant 100 Spectrophotometer</td><td>General Electric</td><td>28-9182-04</td><td></td></tr><tr><td>Sodium thiosulfate</td><td>Fisher Chemical</td><td>S445-3</td><td></td></tr><tr><td>Membrane filter (47 mm, 0.45 &amp;micro;m)</td><td>EMD Millipore</td><td>HAWP04700</td><td></td></tr><tr><td>m-ColiBlue24 broth media</td><td>EMD Millipore</td><td>M00PMCB24</td><td></td></tr><tr><td>Petri dish with pad (47 mm)</td><td>Fisherbrand</td><td>09-720-500</td><td></td></tr><tr><td>Vacuum Manifold</td><td>Thermo Scientific/Nalgene</td><td>09-752-5</td><td></td></tr><tr><td>Filter funnels</td><td>Thermo Scientific/Nalgene</td><td>09-747</td><td></td></tr><tr><td>Pseudomonas syringae</td><td>HER</td><td>1102</td><td></td></tr><tr><td>Phosphate Buffered Saline</td><td>Thermo Scientific</td><td>10010031</td><td>Solution may also be mixed from source compounds according to any basic recipe</td></tr></tbody></table>

a method to test the efficacy of handwashing for the removal of emerging infectious pathogens

La lavata a mano è ampiamente raccomandata per prevenire la trasmissione delle malattie infettive. Tuttavia, esistono scarse prove comparabili sull&#39;efficacia dei metodi di lavaggio a mano in generale. Inoltre, esistono poche prove che consentono di confrontare i metodi di lavaggio a mano per determinare quali sono i più efficaci per la rimozione di patogeni infettivi. È necessaria la ricerca per fornire prove sui diversi approcci di lavaggio a mano che possono essere utilizzati durante i focolai di malattie infettive. Qui viene descritto un metodo di laboratorio per valutare l&#39;efficacia dei metodi di lavaggio a mano per rimuovere i microrganismi dalle mani e la loro persistenza nell&#39;acqua di risciacquo. Le mani dei volontari sono dapprima forate con l&#39;organismo di prova e poi lavate con ogni metodo di lavaggio a mano d&#39;interesse. In generale, i microrganismi surrogati vengono utilizzati per proteggere i soggetti umani dalla malattia. Il numero di organismi che rimangono sulle mani dei volontari dopo il lavaggio viene testato utilizzando un metodo &quot;succo di guanti&quot; modificato: le mani vengono collocate in guanti con un eluE vengono sciacquati per sospendere i microrganismi e renderli disponibili per l&#39;analisi mediante filtrazione a membrana (batteri) o test di placca (virus / batteriofagi). L&#39;acqua di risciacquo prodotta dalla lavata a mano viene direttamente raccolta per l&#39;analisi. L&#39;efficacia del lavaggio manuale viene quantificata confrontando il valore di riduzione del log tra i campioni prelevati dopo lavaggio a campioni senza lavaggio a mano. La persistenza della risciacquatura dell&#39;acqua è quantificata confrontando i campioni di acqua di risciacquo da vari metodi di lavaggio a mano a campioni prelevati dopo lavaggio a mano con acqua sufficiente. Mentre questo metodo è limitato dalla necessità di utilizzare organismi surrogati per preservare la sicurezza dei volontari umani, raccoglie aspetti di lavaggio a mano che sono difficili da replicare in uno studio in vitro e riempie le lacune di ricerca sull&#39;efficacia della lavaggio e la persistenza degli organismi infettivi in ​​risciacquo acqua.

Qui, il protocollo ( Figura 1 ) è stato completato con 18 volontari, che sono stati testati sia con E. coli che Phi6. Differenze significative sono state riscontrate tra i risultati di lavaggio a mano con E. coli sia con e senza carico del suolo e Phi6 con carico del suolo ( Figura 2 e Figura 3 ). Per E. coli senza carico del suolo, la lavata a mano con HTH, NaDCC e NaOCl stabilizzata ha portato ad una riduzione significativa di log rispetto alla lavaggio a mano solo con acqua (F (6,102) = 2,72, p = 0,034). Con il carico del suolo, HTH ha portato ad una riduzione significativa di log di E. coli rispetto all&#39;acqua solo, HWWS e ABHS (F (6,102) = 3,94, p &lt;0,001). Non esisteva alcuna differenza significativa tra i metodi per Phi6 senza carico del suolo (F (6,66) = 2,04, p = 0,073). Tuttavia, per Phi6 con carico del suolo (F (6,102) = 7,01, p &lt;0,001), l&#39;acqua da sola ha prodotto un greRiduzione del log di ABHS o stabilizzato NaOCl e HWWS in una riduzione più grande di log di ABHS, stabilizzato NaOCl e generato NaOCl. HTH aveva anche una riduzione più grande di log rispetto a ABHS e stabilizzato NaOCl, e NaDCC ha portato ad una riduzione maggiore del log rispetto a NaOCl e ABHS stabilizzati. Mentre HTH è stato eseguito in modo più coerente nelle condizioni, avremmo cautela nei confronti dell&#39;eccessiva interpretazione di risultati significativi, in quanto molti intervalli di confidenza sono stati grandi, che vanno da meno di 0,5 log a più di 1,5 log reduction in molti casi. Nell&#39;acqua di risciacquo, il cloro ha determinato una riduzione significativa di log di E. coli persistente nell&#39;acqua di risciacquo rispetto a HWWS (senza carico del suolo, F (4,68) = 331,7, p &lt;0,001; con carico del terreno, F (4,68 ) = 162,44, p &lt;0,001) ( Figura 4 ). Questo stesso modello è stato trovato in Phi6 senza carico del suolo ((F (4,43) = 8,95, P &lt;0,001), con tutte le soluzioni di cloro con conseguenteUna riduzione nettamente maggiore di Phi6 nell&#39;acqua di risciacquo che HWWS. Non esistono differenze significative nella persistenza nell&#39;acqua di risciacquo con Phi6 e carico del terreno ((F (4,67) = 3,35, p = 0,071) ( Figura 5 ). <img alt="Figura 1" class="xfigimg" src="/files/ftp_upload/55604/55604fig1.jpg" /> Figura 1: Panoramica sull&#39;esperimento. Le cinque fasi intraprese per ciascun ciclo di lavaggio a mano comprendono: 1) test di pH, 2) inoculazione delle mani, 3) lavaggio a mano, 4) lavaggio delle mani e 5) decontaminazione delle mani per ognuna delle otto condizioni esaminate. <a href="http://ecsource.jove.com/files/ftp_upload/55604/55604fig1large.jpg" target="_blank">Clicca qui per visualizzare una versione più grande di questa figura.</a> <img alt="figura 2" class="xfigimg" src="/files/ftp_upload/55604/55604fig2.jpg" /> Figura 2: lavaggio a mano di E. coli<html> ULT. Rispetto a nessun lavaggio a mano, i metodi di lavaggio a mano hanno provocato una riduzione media di log in E. coli di 1,94-3,01 senza carico del suolo e 2,18-3,34 con carico del suolo. La lavata a mano con acqua ha dimostrato la minima riduzione di E. coli in entrambe le condizioni (1,94 e 2,18 log). La lavata a mano con NaDCC ha determinato la massima riduzione senza carico del terreno (3,01) e HTH ha portato la più grande riduzione con carico del terreno (3,34). Nei grafici, la riga rappresenta la riduzione percentuale degli organismi e le barre di errore rappresentano l&#39;errore standard della riduzione del registro. Ctrl B, controllo B; HWWS, lavaggio a mano con sapone; ABHS, sanitizzatore manuale a base di alcool; HTH, ipoclorito ad alta prova; NaDCC, sodio dicloroisocianurato; St NaOCl, ipoclorito sodico stabilizzato; Gen NaOCl, generò ipoclorito di sodio. <a href="http://ecsource.jove.com/files/ftp_upload/55604/55604fig2large.jpg" target="_blank">Clicca qui per visualizzare una versione più grande di questa figura.</a> Ve_content &quot;per: keep-together.within-page =&quot; 1 &quot;&gt; <img alt="Figura 3" class="xfigimg" src="/files/ftp_upload/55604/55604fig3.jpg" /> Figura 3: risultati di lavaggio a mano di Phi6. Rispetto a nessun lavaggio a mano, i metodi di lavaggio a mano hanno provocato una riduzione media di log in Phi6 di 2,44-3,06 senza carico del suolo e 2,71-3,69 con carico del suolo. La lavata a mano con sapone ha dimostrato la minima riduzione di Phi6 senza carico del suolo (2,44) e lavaggio a mano con NaOCl stabilizzato ha portato alla riduzione minima con carico del terreno (2,71). La lavaggio a mano con NaOCl generato ha portato alla riduzione massima senza carico del terreno (3,06) e la lavaggio a mano con sapone ha portato alla riduzione più grande con carico del terreno (3,69). Nei grafici, la riga rappresenta la riduzione percentuale degli organismi e le barre di errore rappresentano l&#39;errore standard della riduzione del registro. Ctrl B, controllo B; HWWS, lavaggio a mano con sapone; ABHS, sanitizzatore manuale a base di alcool; HTH, ipoclorito ad alta prova; NaDCC, sodio dicloroisocianurato;St NaOCl, ipoclorito sodico stabilizzato; Gen NaOCl, generò ipoclorito di sodio. <a href="http://ecsource.jove.com/files/ftp_upload/55604/55604fig3large.jpg" target="_blank">Clicca qui per visualizzare una versione più grande di questa figura.</a> <img alt="Figura 4" class="xfigimg" src="/files/ftp_upload/55604/55604fig4.jpg" /> Figura 4: risultati di risciacquo a mano di E. coli . Rispetto al lavaggio a mano solo con acqua, la riduzione media del tronco di E. coli rimanente nell&#39;acqua di risciacquo è stata di 0,28-4,77 senza carico del suolo e 0,21-4,49 con carico del terreno. Sia con e senza carico del suolo, la più piccola riduzione è stata trovata nella lavaggio a mano con sapone (0,28 e 0,21). Le maggiori riduzioni sono state osservate con NaOCl stabilizzato e generato senza carico del suolo (entrambi 4.77) e con HTH e generato NaOCl con carico del terreno. Nei grafici, la riga rappresenta la percentuale di riduzione degli organismi e le barre di errore rappresentano le sErrore tandard della riduzione del registro. HWWS, lavaggio a mano con sapone; ABHS, sanitizzatore manuale a base di alcool; HTH, ipoclorito ad alta prova; NaDCC, sodio dicloroisocianurato; St NaOCl, ipoclorito sodico stabilizzato; Gen NaOCl, generò ipoclorito di sodio. <a href="http://ecsource.jove.com/files/ftp_upload/55604/55604fig4large.jpg" target="_blank">Clicca qui per visualizzare una versione più grande di questa figura.</a> <img alt="Figura 5" class="xfigimg" src="/files/ftp_upload/55604/55604fig5.jpg" /> Figura 5: risultati di risciacquo a mano di Phi6. Rispetto al lavaggio a mano solo con acqua, la riduzione media di log del Phi6 rimanente nell&#39;acqua di risciacquo è stata di 1,26-2,02 senza carico del suolo e 1,30-2,20 con carico del terreno. Con il carico del suolo, la riduzione minima è stata trovata nella lavaggio a mano con sapone (1,26). Senza carico del terreno, HTH ha portato alla riduzione minima (2,02). Le maggiori riduzioni sono state osservate con e senza carico del suolo con NaDCC(2,02 e 2,20). Nei grafici, la riga rappresenta la riduzione percentuale degli organismi e le barre di errore rappresentano l&#39;errore standard della riduzione del registro. HWWS, lavaggio a mano con sapone; ABHS, sanitizzatore manuale a base di alcool; HTH, ipoclorito ad alta prova; NaDCC, sodio dicloroisocianurato; St NaOCl, ipoclorito sodico stabilizzato; Gen NaOCl, generò ipoclorito di sodio. <a href="http://ecsource.jove.com/files/ftp_upload/55604/55604fig5large.jpg" target="_blank">Clicca qui per visualizzare una versione più grande di questa figura.</a>

Watch this Scientific Journal Video about A Method to Test the Efficacy of Handwashing for the Removal of Emerging Infectious Pathogens at JoVE.com

A Method to Test the Efficacy of Handwashing for the Removal of Emerging Infectious Pathogens

La lavata a mano è ampiamente raccomandata per prevenire la trasmissione delle malattie infettive. Tuttavia, non vi è alcuna prova su cui i metodi di lavaggio della mano sono più efficaci nella rimozione di patogeni delle malattie infettive. Abbiamo sviluppato un metodo per valutare l&#39;efficacia dei metodi di lavaggio a mano alla rimozione dei microrganismi.

Un metodo per testare l&#39;efficacia della lavata a mano per la rimozione di nuovi patogeni infettivi

Handwashing is widely recommended to prevent infectious disease transmission. However, little comparable evidence exists on the efficacy of handwashing ...

a-method-to-test-efficacy-handwashing-for-removal-emerging-infectious

Nanyang Technological University

Research

JoVE Journal

Immunology and Infection

19.0K Views.  Tufts University.  La lavata a mano è ampiamente raccomandata per prevenire la trasmissione delle malattie infettive. Tuttavia, non vi è alcuna prova su cui i metodi di lavaggio della mano sono più efficaci nella rimozione di patogeni delle malattie infettive. Abbiamo sviluppato un metodo per valutare l'efficacia dei metodi di lavaggio a mano alla rimozione dei microrganismi. 

Video: A Method to Test the Efficacy of Handwashing for the Removal of Emerging Infectious Pathogens

EPA Method 1615. Measurement of Enterovirus and Norovirus Occurrence in Water by Culture and RT-qPCR. I. Collection of Virus Samples

EPA Method 1615 was developed with a goal of providing a standard method for measuring enteroviruses and noroviruses in environmental and drinking waters. The standardized sampling component of the method concentrates viruses that may be present in water by passage of a minimum specified volume of water through an electropositive cartridge filter. The minimum specified volumes for surface and finished/ground water are 300 L and 1,500 L, respectively. A major method limitation is the tendency for the filters to clog before meeting the sample volume requirement. Studies using two different, but equivalent, cartridge filter options showed that filter clogging was a problem with 10% of the samples with one of the filter types compared to 6% with the other filter type. Clogging tends to increase with turbidity, but cannot be predicted based on turbidity measurements only. From a cost standpoint one of the filter options is preferable over the other, but the water quality and experience with the water system to be sampled should be taken into consideration in making filter selections.

EPA Method 1615 uses an electropositive filter to concentrate enteroviruses and noroviruses in environmental and drinking waters. This manuscript describes the procedure for collecting samples for Method 1615 analyses.

Metodo EPA 1615. Misura di Enterovirus e Norovirus Evento in acqua per la cultura e RT-qPCR. I. Raccolta di campioni di virus

EPA Method 1615 was developed with a goal of providing a standard method for measuring enteroviruses and noroviruses in environmental and drinking waters. ...

Ricerca

Ambiente

Determining Viral Disinfection Efficacy of Hot Water Laundering

This protocol provides an example of a laboratory process for conducting laundering studies that generate data on viral disinfection. While the protocol was developed for research during the coronavirus disease 2019 (COVID-19) pandemic, it is intended to be a framework, adaptable to other virus disinfection studies; it demonstrates the steps for preparing the test virus, inoculating the test material, assessing visual and integrity changes to the washed items due to the laundering process, and quantifying the reduction in viral load. Additionally, the protocol outlines the necessary quality control samples for ensuring the experiments are not biased by contamination and measurements/observations that should be recorded to track the material integrity of the personal protective equipment (PPE) items after multiple laundering cycles. The representative results presented with the protocol use the Phi6 bacteriophage inoculated onto cotton scrub, denim, and cotton face-covering materials and indicate that the hot water laundering and drying process achieved over a 3-log (99.9%) reduction in viral load for all samples (a 3-log reduction is the disinfectant performance metric in U.S. Environmental Protection Agency's Product Performance Test Guideline 810.2200). The reduction in viral load was uniform across different locations on the PPE items. The results of this viral disinfection efficacy testing protocol should help the scientific community explore the effectiveness of home laundering for other types of test viruses and laundering procedures.

In response to the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic, a laboratory protocol was developed to test the viral disinfection efficacy of hot water laundering of cloth face coverings, cotton scrubs, and denim pants. The Phi6 virus (bacteriophage) was used as the organism to test disinfection efficacy.

Determinazione dell'efficacia della disinfezione virale del lavaggio dell'acqua calda

This protocol provides an example of a laboratory process for conducting laundering studies that generate data on viral disinfection. While the protocol ...

Ingegneria

Antimicrobial Synergy Testing by the Inkjet Printer-assisted Automated Checkerboard Array and the Manual Time-kill Method

As rates of multidrug-resistant (MDR) pathogens continue to rise, outpacing the development of new antimicrobials, novel approaches to treatment of MDR bacteria are increasingly becoming a necessity. One such approach is combination therapy, in which two or more antibiotics are used together to treat an infection against which one or both of the drugs may be ineffective alone. When two drugs, in combination, exert a greater than additive effect, they are considered synergistic. In vitro investigation of synergistic activity is an important first step in evaluating the possible efficacy of drug combinations. Two main in vitro synergy testing methods have been developed: the checkerboard array and the time-kill study. In this paper, we present an automated checkerboard array method that makes use of inkjet printing technology to increase the efficiency and accuracy of this technique, as well as a standard manual time-kill synergy method. The automated checkerboard array can serve as a high-throughput screening assay, while the manual time-kill study provides additional, complementary data on synergistic activity and killing.
The checkerboard array is a modification of standard minimum inhibitory concentration (MIC) testing, in which bacteria are incubated with antibiotics at different concentration combinations and evaluated for growth inhibition after overnight incubation. Manual performance of the checkerboard array requires a laborious and error-prone series of calculations and dilutions. In the automated method presented here, the calculation and dispensing of required antibiotic stock solution volumes are automated through the use of inkjet printer technology. In the time-kill synergy assay, bacteria are incubated with the antibiotics of interest, both together and individually, and sampled at intervals over the course of 24 h for quantitative culture. The results can determine whether a combination is synergistic and whether it is bactericidal, and provide data on inhibition and killing of bacteria over time.

Antimicrobial synergy testing is used to evaluate the effect of two or more antibiotics used in combination and is typically performed by one of two methods: the checkerboard array or the time-kill assay. Here, we present an automated, inkjet printer-assisted checkerboard array synergy technique and a classic time-kill synergy study.

Antimicrobica sinergia test di stampante a getto d'inchiostro-assistita Checkerboard automatizzato matrice e il metodo di Time-kill manuale

As rates of multidrug-resistant (MDR) pathogens continue to rise, outpacing the development of new antimicrobials, novel approaches to treatment of MDR ...

Immunologia e infezioni

Prion Safety Laboratory Swipe Test

Transmission of iatrogenic prion disease has occurred from contaminated neurosurgical tools, transplant materials, and occupational exposure to prion-contaminated laboratory tools. Prions cause disease by the templated misfolding of the normal cellular form of the prion protein, PrPC, into the misfolded and pathogenic form PrPSc and are invariably fatal. Reducing iatrogenic and occupational prion transmission is challenging. First, prions can bind to and persist on surfaces for long periods of time. Second, prions are highly resistant to inactivation. Given this, surfaces can retain infectivity for long periods of time following ineffective decontamination. Not only can this pose a potential occupational risk for prion laboratory workers, but it could potentially cross-contaminate laboratory experiments utilizing sensitive prion amplification techniques. The protocol described here for a prion safety laboratory swipe test includes steps for the identification and documentation of high-traffic laboratory areas, recommended swabbing controls to ensure the validity of results, steps to identify proper responses to positive surface swabbing sites, representative results from prion swipe testing, as well as potential artifactual results. Overall, the prion safety laboratory swipe test can be implemented as part of a broader prion safety program to assess decontamination of surfaces, monitor common spaces for prion contamination, and implement the documentation of prion decontamination status.

A method to assess commonly used areas in laboratory settings for prion contamination and effective decontamination is lacking. The protocol described here provides key fundamentals for implementing a laboratory prion safety swipe test that can easily be modified to meet the individual needs of specific laboratories.

Test di strisciamento del laboratorio di sicurezza dei prioni

Transmission of iatrogenic prion disease has occurred from contaminated neurosurgical tools, transplant materials, and occupational exposure to ...

Swabbing the Urban Environment - A Pipeline for Sampling and Detection of SARS-CoV-2 From Environmental Reservoirs

To control community transmission of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) during the 2020 global pandemic, most countries implemented strategies based on direct human testing, face covering, and surface disinfection. Under the assumption that the main route of transmission includes aerosols and respiratory droplets, efforts to detect SARS-CoV-2 in fomites have focused on locations suspected of high prevalence (e.g., hospital wards, cruise ships, and mass transportation systems). To investigate the presence of SARS-CoV-2 on surfaces in the urban environment that are rarely cleaned and&#160;seldomly disinfected, 350 citizens were enlisted from the greater San Diego County. In total, these citizen scientists collected 4,080 samples. An online platform was developed to monitor sampling kit delivery and pickup, as well as to collect sample data. The sampling kits were mostly built from supplies available in pandemic-stressed stores. Samples were processed using reagents that were easy to access despite the recurrent supply shortage. The methods used were highly sensitive and resistant to inhibitors that are commonly present in environmental samples. The proposed experimental design and processing methods were successful at engaging numerous citizen scientists who effectively gathered samples from diverse surface areas. The workflow and methods described here are relevant to survey the urban environment for other viruses, which are of public health concern and pose a threat for&#160;future pandemics.

A citizen science project was designed to recruit San Diego residents to collect environmental samples for SARS-CoV-2. A multilingual web-based platform was created for data submission using a user-friendly mobile device interface. A laboratory information management system facilitated the collection of thousands of geographically diverse samples with real-time outcome tracking.

Tamponare l'ambiente urbano - Una pipeline per il campionamento e il rilevamento della SARS-CoV-2 dai bacini ambientali

To control community transmission of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) during the 2020 global pandemic, most countries ...

Biologia

Evaluation of the Efficacy of Organic Peroxyacids for Eradicating Dairy Biofilms Using an Approach Combining Static and Dynamic Methods

The presence of biofilms in the dairy industry is of major concern, as they may lead to the production of unsafe and altered dairy products due to their high resistance to most clean-in-place (CIP) procedures frequently used in processing plants. Therefore, it is imperative to develop new biofilm control strategies for the dairy industry. This protocol is aimed at evaluating the efficacy of organic peroxyacids (peracetic, perpropionic, and perlactic acids and a commercial peracetic acid-based disinfectant) for eradicating dairy biofilms using a combination of static and dynamic methods. All the disinfectants were tested on the strongest biofilm-producing bacteria in either a single or a mixed biofilm using the minimum biofilm eradication concentration (MBEC) assay, a static high-throughput screening method. A contact time of 5 min with the disinfectants at the recommended concentrations successfully eradicated both the single and mixed biofilms. Studies are currently ongoing to confirm these observations using the Center for Disease Control (CDC) biofilm reactor, a dynamic method to mimic in situ conditions. This type of bioreactor enables the use of a stainless-steel surface, which constitutes most industrial equipment and surfaces. The preliminary results from the reactor appear to confirm the efficacy of organic peroxyacids against biofilms. The combined approach described in this study may be used to develop and test new biological or chemical formulations for controlling biofilms and eradicating microorganisms.

This protocol describes an approach combining static and dynamic methods to evaluate the efficacy of organic peroxyacids for eradicating biofilms in the dairy industry. This approach may also be used to test the effectiveness of new biological or chemical formulations for controlling biofilms.

Valutazione dell'efficacia dei perossiacidi organici per l'eradicazione dei biofilm lattiero-caseari utilizzando un approccio che combina metodi statici e dinamici

The presence of biofilms in the dairy industry is of major concern, as they may lead to the production of unsafe and altered dairy products due to their ...

Efficacia antimicrobica dei tessuti rivestiti con nano-erbe

An Antimicrobial Fabric Using Nano-Herbal Encapsulation of Essential Oils

Lab coats are widely used in biohazard laboratories and healthcare facilities as protective garments to prevent direct exposure to pathogens, spills, and burns. These cotton-based protective coats provide ideal conditions for microbial growth and attachment sites due to their porous nature, moisture-holding capacity, and retention of warmth from the user&#39;s body. Several studies have demonstrated the survival of pathogenic bacteria on hospital garments and lab coats, acting as vectors of microbial transmission.
A common approach to fix these problems is the application of antimicrobial agents in textile finishing, but concerns have been raised due to the toxicity and environmental effects of many synthetic chemicals. The ongoing pandemic has also opened a window for the investigation of effective antimicrobials and eco-friendly and toxic-free formulations. This study uses two natural bioactive compounds, carvacrol and thymol, encapsulated in chitosan nanoparticles, which guarantee effective protection against four human pathogens with up to a 4-log reduction (99.99%). These pathogens are frequently detected in lab coats used in biohazard laboratories.
The treated fabrics also resisted up to 10 wash cycles with 90% microbial reduction, which is sufficient for the intended use. We made modifications to the existing standard fabric tests to better represent the typical scenarios of lab coat usage. These refinements allow for a more accurate evaluation of the effectiveness of antimicrobial lab coats and for the simulation of the fate of any accidental microbial spills that must be neutralized within a short time. Further studies are recommended to investigate the accumulation of pathogens over time on antimicrobial lab coats compared to regular protective coats.

Autore in primo piano: Un tessuto antimicrobico che utilizza l'incapsulamento nano-erboristico di oli essenziali  

Antimicrobial lab coats prevent the cross-contamination of pathogen accumulation and accidental bio-spills. Here, we describe the protocol for developing a skin-friendly antimicrobial fabric using nano-herbal encapsulation and modified standard tests to precisely evaluate the efficacy and suitability for typical usage of the lab coat.

Un tessuto antimicrobico che utilizza l'incapsulamento nano-erboristico di oli essenziali

Lab coats are widely used in biohazard laboratories and healthcare facilities as protective garments to prevent direct exposure to pathogens, spills, and ...

A Deferred Growth Inhibition Assay to Evaluate the Effect of Bacteria-Derived Antimicrobial Compounds

Source: Moran, J. C. et al., <a href="https://app.jove.com/v/54437">Deferred Growth Inhibition Assay to Quantify the Effect of Bacteria-derived Antimicrobials on Competition.</a> J. Vis. Exp.&#160;(2016)
This video showcases the deferred growth inhibition assay, used to investigate how antimicrobial compounds produced by one type of bacteria affect another competing bacterial strain.

Un saggio di inibizione della crescita differita per valutare l'effetto dei composti antimicrobici derivati da batteri

The following method is adapted&#160;to test competitor strains with inhibitor-producing strains.
NOTE: This protocol involves aerosol generation with ...

Un metodo per testare l'efficacia della lavata a mano per la rimozione di nuovi patogeni infettivi

Riepilogo

Esplora Altri Video

Capitoli in questo video

Metodo EPA 1615. Misura di Enterovirus e Norovirus Evento in acqua per la cultura e RT-qPCR. I. Raccolta di campioni di virus

Determinazione dell'efficacia della disinfezione virale del lavaggio dell'acqua calda

Antimicrobica sinergia test di stampante a getto d'inchiostro-assistita Checkerboard automatizzato matrice e il metodo di Time-kill manuale

Test di strisciamento del laboratorio di sicurezza dei prioni

Tamponare l'ambiente urbano - Una pipeline per il campionamento e il rilevamento della SARS-CoV-2 dai bacini ambientali

Valutazione dell'efficacia dei perossiacidi organici per l'eradicazione dei biofilm lattiero-caseari utilizzando un approccio che combina metodi statici e dinamici

Un tessuto antimicrobico che utilizza l'incapsulamento nano-erboristico di oli essenziali

Un tessuto antimicrobico che utilizza l'incapsulamento nano-erboristico di oli essenziali

Un tessuto antimicrobico che utilizza l'incapsulamento nano-erboristico di oli essenziali

Un saggio di inibizione della crescita differita per valutare l'effetto dei composti antimicrobici derivati da batteri