Tracing ΦX174 bacteriophage spreading during aerosol‑generating procedures in a dental clinic
Objective The aim of this study was to test the plausibility of using the ΦX174 bacteriophage as a tracer of viral aerosols spreading in a dental aerosol-generating procedure (AGP) model. Methods ΦX174 bacteriophage (~ 108 plaque-forming units (PFU)/mL) was added into instrument irrigation reservoir...
- Autores:
-
Beltrán, Edgar O.
Castellanos, Jaime E.
Corredor, Zayda L.
Morgado, Wendy
Zarta, Olga L.
Cortés, Andrea
Avila, Viviana
Martignon, Stefania
- Tipo de recurso:
- Article of investigation
- Fecha de publicación:
- 2023
- Institución:
- Corporación Universidad de la Costa
- Repositorio:
- REDICUC - Repositorio CUC
- Idioma:
- eng
- OAI Identifier:
- oai:repositorio.cuc.edu.co:11323/10469
- Acceso en línea:
- https://hdl.handle.net/11323/10469
https://repositorio.cuc.edu.co/
- Palabra clave:
- Bacteriophage
Aerosol
Aerosol-generating procedure
Dental setting
Virus laden aerosols
Airborne transmission
- Rights
- embargoedAccess
- License
- Atribución 4.0 Internacional (CC BY 4.0)
| id |
RCUC2_b66b6ac0f4f0ade217053e795c0a0b7d |
|---|---|
| oai_identifier_str |
oai:repositorio.cuc.edu.co:11323/10469 |
| network_acronym_str |
RCUC2 |
| network_name_str |
REDICUC - Repositorio CUC |
| repository_id_str |
|
| dc.title.eng.fl_str_mv |
Tracing ΦX174 bacteriophage spreading during aerosol‑generating procedures in a dental clinic |
| title |
Tracing ΦX174 bacteriophage spreading during aerosol‑generating procedures in a dental clinic |
| spellingShingle |
Tracing ΦX174 bacteriophage spreading during aerosol‑generating procedures in a dental clinic Bacteriophage Aerosol Aerosol-generating procedure Dental setting Virus laden aerosols Airborne transmission |
| title_short |
Tracing ΦX174 bacteriophage spreading during aerosol‑generating procedures in a dental clinic |
| title_full |
Tracing ΦX174 bacteriophage spreading during aerosol‑generating procedures in a dental clinic |
| title_fullStr |
Tracing ΦX174 bacteriophage spreading during aerosol‑generating procedures in a dental clinic |
| title_full_unstemmed |
Tracing ΦX174 bacteriophage spreading during aerosol‑generating procedures in a dental clinic |
| title_sort |
Tracing ΦX174 bacteriophage spreading during aerosol‑generating procedures in a dental clinic |
| dc.creator.fl_str_mv |
Beltrán, Edgar O. Castellanos, Jaime E. Corredor, Zayda L. Morgado, Wendy Zarta, Olga L. Cortés, Andrea Avila, Viviana Martignon, Stefania |
| dc.contributor.author.none.fl_str_mv |
Beltrán, Edgar O. Castellanos, Jaime E. Corredor, Zayda L. Morgado, Wendy Zarta, Olga L. Cortés, Andrea Avila, Viviana Martignon, Stefania |
| dc.subject.proposal.eng.fl_str_mv |
Bacteriophage Aerosol Aerosol-generating procedure Dental setting Virus laden aerosols Airborne transmission |
| topic |
Bacteriophage Aerosol Aerosol-generating procedure Dental setting Virus laden aerosols Airborne transmission |
| description |
Objective The aim of this study was to test the plausibility of using the ΦX174 bacteriophage as a tracer of viral aerosols spreading in a dental aerosol-generating procedure (AGP) model. Methods ΦX174 bacteriophage (~ 108 plaque-forming units (PFU)/mL) was added into instrument irrigation reservoirs and aerosolized during class-IV cavity preparations followed by composite fllings on natural upper-anterior teeth (n=3) in a phantom head. Droplets/aerosols were sampled through a passive approach that consisted of Escherichia coli strain C600 cultures immersed in a LB top agar layer in Petri dishes (PDs) in a double-layer technique. In addition, an active approach consisted of E coli C600 on PDs sets mounted in a six-stage cascade Andersen impactor (AI) (simulating human inhalation). The AI was located at 30 cm from the mannequin during AGP and afterwards at 1.5 m. After collection PDs were incubated overnight (18 h at 37 °C) and bacterial lysis was quantifed. Results The passive approach disclosed PFUs mainly concentrated over the dental practitioner, on the mannequin’s chest and shoulder and up to 90 cm apart, facing the opposite side of the AGP’s source (around the spittoon). The maximum aerosol spreading distance was 1.5 m in front of the mannequin’s mouth. The active approach disclosed collection of PFUs corresponding to stages (and aerodynamic diameters) 5 (1.1–2.1 µm) and 6 (0.65–1.1 µm), mimicking access to the lower respiratory airways. Conclusion The ΦX174 bacteriophage can be used as a traceable viral surrogate in simulated studies contributing to understand dental bioaerosol’s behavior, its spreading, and its potential threat for upper and lower respiratory tract. Clinical relevance The probability to fnd infectious virus during AGPs is high. This suggests the need to continue characterizing the spreading viral agents in diferent clinical settings through combination of passive and active approaches. In addition, subsequent identifcation and implementation of virus-related mitigation strategies is relevant to avoid occupational virus infections. |
| publishDate |
2023 |
| dc.date.accessioned.none.fl_str_mv |
2023-09-11T18:57:50Z |
| dc.date.available.none.fl_str_mv |
2023-09-11T18:57:50Z 2024 |
| dc.date.issued.none.fl_str_mv |
2023 |
| dc.type.spa.fl_str_mv |
Artículo de revista |
| dc.type.coar.spa.fl_str_mv |
http://purl.org/coar/resource_type/c_2df8fbb1 |
| dc.type.content.spa.fl_str_mv |
Text |
| dc.type.driver.spa.fl_str_mv |
info:eu-repo/semantics/article |
| dc.type.redcol.spa.fl_str_mv |
http://purl.org/redcol/resource_type/ART |
| dc.type.version.spa.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
| dc.type.coarversion.spa.fl_str_mv |
http://purl.org/coar/version/c_970fb48d4fbd8a85 |
| format |
http://purl.org/coar/resource_type/c_2df8fbb1 |
| status_str |
publishedVersion |
| dc.identifier.citation.spa.fl_str_mv |
Beltrán, E.O., Castellanos, J.E., Corredor, Z.L. et al. Tracing ΦX174 bacteriophage spreading during aerosol-generating procedures in a dental clinic. Clin Oral Invest 27, 3221–3231 (2023). https://doi.org/10.1007/s00784-023-04937-z |
| dc.identifier.issn.spa.fl_str_mv |
1432-6981 |
| dc.identifier.uri.none.fl_str_mv |
https://hdl.handle.net/11323/10469 |
| dc.identifier.doi.none.fl_str_mv |
10.1007/s00784-023-04937-z |
| dc.identifier.eissn.spa.fl_str_mv |
1436-3771 |
| dc.identifier.instname.spa.fl_str_mv |
Corporación Universidad de la Costa |
| dc.identifier.reponame.spa.fl_str_mv |
REDICUC - Repositorio CUC |
| dc.identifier.repourl.spa.fl_str_mv |
https://repositorio.cuc.edu.co/ |
| identifier_str_mv |
Beltrán, E.O., Castellanos, J.E., Corredor, Z.L. et al. Tracing ΦX174 bacteriophage spreading during aerosol-generating procedures in a dental clinic. Clin Oral Invest 27, 3221–3231 (2023). https://doi.org/10.1007/s00784-023-04937-z 1432-6981 10.1007/s00784-023-04937-z 1436-3771 Corporación Universidad de la Costa REDICUC - Repositorio CUC |
| url |
https://hdl.handle.net/11323/10469 https://repositorio.cuc.edu.co/ |
| dc.language.iso.spa.fl_str_mv |
eng |
| language |
eng |
| dc.relation.ispartofjournal.spa.fl_str_mv |
Clinical Oral Investigations |
| dc.relation.references.spa.fl_str_mv |
1. Ren Y, Feng C, Rasubala L, Malmstrom H, Eliav E (2020) Risk for dental healthcare professionals during the COVID-19 global pandemic: an evidence-based assessment. J Dent 101:103434. https://doi.org/10.1016/j.jdent.2020.103434 2. Odeh ND, Babkair H, Abu-Hammad S, Borzangy S, AbuHammad A, Abu-Hammad O (2020) COVID-19: present and future challenges for dental practice. Int J Environ Res Public Health 17(9):3151. https://doi.org/10.3390/ijerph17093151 3. World Health Organization (2020) Transmission of SARS-CoV-2: implications for infection prevention precautions. Available at: https://www.who.int/emergencies/diseases/novel-coronavirus2019?gclid=Cj0KCQjw0YD4BRD2ARIsAHwmKVmNyTZntqH Y8mnWiIWPlKpj41bEGY-UjmYkbJ-6mjfBTYlMQrrQLV8aAtD REALw_wcB. Accessed June 2023 4. Meng L, Hua F, Bian Z (2020) Coronavirus disease 2019 (COVID-19): emerging and future challenges for dental and oral medicine. J Dent Res 99(5):481–487. https://doi.org/10.1177/ 0022034520914246 5. Allison JR, Dowson C, Jakubovics NS, Nile C, Durham J, Holliday R (2022) Waterline disinfectants reduce dental bioaerosols: a multitracer validation. J Dent Res 101(10):1198–1204. https:// doi.org/10.1177/00220345221093522 6. Bourouiba L (2022) Turbulent gas clouds and respiratory pathogen emissions: potential implications for reducing transmission of COVID-19. JAMA 323(18):1837–1838. https://doi.org/10.1001/ jama.2020.4756 7. Zemouri C, Volgenant CMC, Buijs MJ, Crielaard W, Rosema NAM, Brandt BW, Laheij AMGA, De Soet JJ (2020) Dental aerosols: microbial composition and spatial distribution. J Oral Microbiol 12(1):1762040. https://doi.org/10.1080/20002297. 2020.1762040 8. Kumbargere Nagraj S, Eachempati P, Paisi M, Nasser M, Sivaramakrishnan G, Verbeek JH (2020) Interventions to reduce contaminated aerosols produced during dental procedures for preventing infectious diseases. Cochrane Database Syst Rev 10(10):CD013686. https://doi.org/10.1002/14651 858.CD013686.pub2 9. Allison JR, Currie CC, Edwards DC, Bowes C, Coulter J, Pickering K, Kozhevnikova E, Durham J, Nile CJ, Jakubovics N, Rostami N, Holliday R (2021) Evaluating aerosol and splatter following dental procedures: addressing new challenges for oral health care and rehabilitation. J Oral Rehabil 48(1):61–72. https://doi.org/10.1111/joor.13098 10. Holliday R, Allison JR, Currie CC, Edwards DC, Bowes C, Pickering K, Reay S, Durham J, Lumb J, Rostami N, Coulter J, Nile C, Jakubovics N (2021) Evaluating contaminated dental aerosol and splatter in an open plan clinic environment: implications for the COVID-19 pandemic. J Dent 105:103565. https:// doi.org/10.1016/j.jdent.2020.103565 11. Llandro H, Allison JR, Currie CC, Edwards DC, Bowes C, Durham J, Jakubovics N, Rostami N, Holliday R (2021) Evaluating splatter and settled aerosol during orthodontic debonding: implications for the COVID-19 pandemic. Br Dent J 8:1–7. https://doi.org/10.1038/s41415-020-2503-9 12. Din AR, Hindocha A, Patel T, Sudarshan S, Cagney N, Koched A, Mueller JD, Seoudi N, Morgan C, Shahdad S, Fleming PS (2020). Quantitative analysis of particulate matter release during orthodontic procedures: a pilot study. Br Dent J 12:1–7. https://doi.org/10.1038/s41415-020-2280-5 13. Timmerman MF, Menso L, Steinfort J, van Winkelhof AJ, van der Weijden GA (2004) Atmospheric contamination during ultrasonic scaling. J Clin Periodontol 31(6):458–462. https:// doi.org/10.1111/j.1600-051X.2004.00511.x 14. Rautemaa R, Nordberg A, Wuolijoki-Saaristo K, Meurman JH (2006) Bacterial aerosols in dental practice – a potential hospital infection problem? J Hosp Infect 64(1):76–81. https://doi. org/10.1016/j.jhin.2006.04.011 15. Holloman JL, Mauriello SM, Pimenta L, Arnold RR (2015) Comparison of suction device with saliva ejector for aerosol and spatter reduction during ultrasonic scaling. J Am Dent Assoc 146(1):27–33. https://doi.org/10.1016/j.adaj.2014.10.001 16. Bennett AM, Fulford MR, Walker JT, Bradshaw DJ, Martin MV, Marsh PD (2020) Microbial aerosols in general dental practice. Br Dent J 189(12):664–667. https://doi.org/10.1038/sj.bdj.4800859 17. Dutil S, Meriaux A, de Latremoille MC, Lazure L, Barbeau J, Duchaine C (2009) Measurement of airborne bacteria and endotoxin generated during dental cleaning. J Occup Environ Hyg 6(2):121–130. https://doi.org/10.1080/15459620802633957 18. King MD, McFarland AR (2012) Use of an Andersen bioaerosol sampler to simultaneously provide culturable particle and culturable organism size distributions. Aerosol Sci Technol 46(8):852–861. https://doi.org/10.1080/02786826.2012.669507 19. Gendron L, Verreault D, Veillette M, Moineau S, Duchaine C (2010) Evaluation of flters for the sampling and quantifcation of RNA phage aerosols. Aerosol Sci Technol 44:893–901. https://doi.org/10.1080/02786826.2010.501351 20. Krupovic M, Bamford DH (2008) Virus evolution: how far does the double beta-barrel viral lineage extend? Nat Rev Microbiol 6(12):941–948. https://doi.org/10.1038/nrmicro2033 21. Verreault D, Moineau S, Duchaine C (2008) Methods for sampling of airborne viruses. Microbiol Mol Biol Rev 72(3):413–444. https://doi.org/10.1128/MMBR.00002-08 22. Verreault D, Rousseau GM, Gendron L, Masse D, Moineau S, Duchaine C (2010) Comparison of polycarbonate and polytetrafuoroethylene flters for sampling of airborne bacteriophages. Aerosol Sci Technol 44:197–201. https://doi.org/10.1080/02786 820903518899 23. Turgeon N, Toulouse MJ, Martel B, Moineau S, Duchaine C (2014) Comparison of fve bacteriophages as models for viral aerosol studies. Appl Environ Microbiol 80(14):4242–4250. https://doi.org/10.1128/AEM.00767-14 24. Appleyard RK (1954) Segregation of new lysogenic types during growth of a doubly lysogenic strain derived from Escherichia coli K12. Genetics 39(4):440–452. https://doi.org/10.1093/genetics/39.4.440 25. Kropinski AM, Mazzocco A, Waddell TE, Lingohr E, Johnson RP (2009) Enumeration of bacteriophages by double agar overlay plaque assay. Methods Mol Biol 501:69–76. https://doi.org/10. 1007/978-1-60327-164-6_7 26. Vernon JJ, Black EVI, Dennis T, Devine DA, Fletcher L, Wood DJ, Nattress BR (2021) Dental mitigation strategies to reduce aerosolization of SARS-CoV-2. J Dent Res 100(13):1461–1467. https://doi.org/10.1177/00220345211032885 27. Rheinbaben F, Schunemann S, Gross T, Wolf MH (2000) Transmission of viruses via contact in a household setting: experiments using bacteriophage straight phiX174 as a model virus. J Hosp Infect 46:61–66 28. Mik G, de Groot I, Gerbrandy JL (1977) Survival of aerosolized bacteriophage phiX174 in air containing ozone-olefn mixtures. J Hyg (London) 78:189–198 29. Dbouk T, Drikakis D (2020) On coughing and airborne droplet transmission to humans. Phys Fluids 32(5):053310. https://doi. org/10.1063/5.0011960 30. Chen L, Zhao J, Peng J, Li X, Deng X, Geng Z, Shen Z, Guo F, Zhang Q, Jin Y, Wang L, Wang S (2020) Detection of SARS-CoV-2 in saliva and characterization of oral symptoms in COVID-19 patients. Cell Prolif 53(12):e12923. https://doi.org/10.1111/cpr.12923 31. Teo AKJ, Choudhury Y, Tan IB, Cher CY, Chew SH, Wan ZY, Cheng LTE, Oon LLE, Tan MH, Chan KS, Hsu LY (2021) Saliva is more sensitive than nasopharyngeal or nasal swabs for diagnosis of asymptomatic and mild COVID-19 infection. Sci Rep 11(1):3134. https://doi.org/10.1038/s41598-021-82787-z. (Erratum.In:SciRep.2021Jun9;11(1):12538) 32. Leung NHL (2021) Transmissibility and transmission of respiratory viruses. Nat Rev Microbiol 19(8):528–545. https://doi.org/ 10.1038/s41579-021-00535-6 33. Gupta G, Mitra D, Ashok KP, Gupta A, Soni S, Ahmed S, Arya A (2014) Efcacy of preprocedural mouth rinsing in reducing aerosol contamination produced by ultrasonic scaler: a pilot study. J Periodontol 85(4):562–568. https://doi.org/10.1902/jop.2013.120616 34. To KK, Tsang OT, Yip CC, Chan KH, Wu TC, Chan JM, Leung WS, Chik TS, Choi CY, Kandamby DH, Lung DC, Tam AR, Poon RW, Fung AY, Hung IF, Cheng VC, Chan JF, Yuen KY (2020) Consistent detection of 2019 novel coronavirus in saliva. Clin Infect Dis 71(15):841–843. https://doi.org/10.1093/cid/ciaa149 35. Paton S, Clark S, Spencer A, Garratt I, Dinesh I, Thompson KA, Bennett A, Pottage T (2022) Characterisation of particle size and viability of SARS-CoV-2 aerosols from a range of nebuliser types using a novel sampling technique. Viruses 19;14(3):639. https:// doi.org/10.3390/v14030639 36. Fröhlich-Nowoisky J, Kampf CJ, Weber B, Hufman JA, Pöhlker C, Andreae MO, Lang-Yona N, Burrows SM, Gunthe SS, Elbert W, Su H, Hoor P, Thines E, Hofmann T, Després VR, Pöschl U (2016) Bioaerosols in the Earth system: climate, health, and ecosystem interactions. Atmos Res 182:346–376, ISSN 0169–8095. https://doi.org/10.1016/j.atmosres.2016.07.018 37. Morgado-Gamero WB, Mendoza Hernandez M, Castillo Ramirez M, Medina-Altahona J, De La Hoz S, Posso Mendoza H, Parody A, Teixeira EC, Agudelo-Castañeda DM (2019) Antibiotic resistance of airborne viable bacteria and size distribution in neonatal intensive care units. Int J Environ Res Public Health 10;16(18):3340. https://doi.org/10. 3390/ijerph16183340 38. Morgado-Gamero WB, Parody A, Medina J, Rodriguez-Villamizar LA, Agudelo-Castañeda D (2021) Multi-antibiotic resistant bacteria in landfll bioaerosols: environmental conditions and biological risk assessment. Environ Pollut 1 290:118037. https://doi.org/10. 1016/j.envpol.2021.118037 39. Broussard IM, Kahwaji CI. Universal Precautions. [Updated 2022 Sep 1]. In: StatPearls [Internet]. Treasure Island (FL): StatPearls Publishing; 2022 Jan-. Available from: https://www.ncbi.nlm.nih. gov/books/NBK470223/ 40. ASHRAE (2020a) COVID-19 (CORONAVIRUS) Preparedness resources. American Society of Heating, Ventilating, and Air- Conditioning Engineers. Available from: https://www.ashrae.org/fle% 20library/technical%20resources/covid-19/in-room-air-cleaner-guida nce-for-reducing-covid-19-in-air-in-your-space-or-room.pdf 41. ASHRAE (2020b) Position Document on Airborne Infectious Diseases, Approved by the Board of Directors, January 19, 2014. Reafrmed by the Technology Council, February 5, 2020. Atlanta, Georgia 42. Occupational Safety and Health Administration (2011) Dentistry workers and employers. Available at https://www.osha.gov/SLTC/ covid-19/dentistry.html. Accessed 28 May 2020 43. Gund M, Isack J, Hannig M, Thieme-Rufng S, Grtner B, Boros G et al (2021) Contamination of surgical mask during aerosolproducing dental treatments. Clin Oral Investig 25:3173–3180. https://doi.org/10.1007/s00784-020-03645-2 44. Balanta-Melo J, Gutiérrez A, Sinisterra G, Díaz-Posso MdM, Gallego D, Villavicencio J, Contreras A (2020) Rubber Dam isolation and high-volume suction reduce ultrafne dental aerosol particles: an experiment in a simulated patient. Appl Sci 10(18):6345. https://doi.org/10.3390/app10186345 45. Morawska L, Tang JW, Bahnfleth W, Bluyssen PM, Boerstra A, Buonanno G, Cao J, Dancer S, Floto A, Franchimon F, Haworth C, Hogeling J, Isaxon C, Jimenez JL, Kurnitski J, Li Y, Loomans M, Marks G, Marr LC, Mazzarella L, Melikov AK, Miller S, Milton DK, Nazaroff W, Nielsen PV, Noakes C, Peccia J, Querol X, Sekhar C, Seppänen O, Tanabe SI, Tellier R, Tham KW, Wargocki P, Wierzbicka A, Yao M (2020) How can airborne transmission of COVID-19 indoors be minimised? Environ Int 142:105832. https://doi.org/10.1016/j.envint.2020. 105832 46. Bai Y, Yao L, Wei T, Tian F, Jin DY, Chen L, Wang M (2020) Presumed asymptomatic carrier transmission of COVID-19. JAMA 323(14):1406–1407. https://doi.org/10.1001/jama.2020. 2565 47. Ren YF, Huang Q, Marzouk T, Richard R, Pembroke K, Martone P, Venner T, Malmstrom H, Eliav E (2020) Efects of mechanical ventilation and portable air cleaner on aerosol removal from dental treatment rooms. J Dent 105:103576. https://doi.org/10. 1016/j.jdent.2020.103576 48. Hiroshi N, Hitoshi O, Tetsuro K, Tomoya S, Tomimasa S, Tamano M, Takaji W, MHLW COVID-19 Response Team, Motoi Suzuki (2020) Closed environments facilitate secondary transmission of coronavirus disease 2019 (COVID-19). medRxiv 2020.02.28.20029272. https:// doi.org/10.1101/2020.02.28.20029272 49. Godwin CC, Batterman SA, Sahni SP, Peng CY (2003) Indoor environment quality in dental clinics: potential concerns from particulate matter. Am J Dent 16(4):260–266 50. Robertson C, Clarkson JE, Aceves-Martins M, Ramsay CR, Richards D, Colloc T, CoDER Working Group (2021) A review of aerosol generation mitigation in international dental guidance. Int Dent J 72(2):203–210. https://doi.org/10.1016/j. identj.2021.04.002 51. Ehtezazi T, Evans DG, Jenkinson ID, Evans PA, Vadgama VJ, Vadgama J, Jarad F, Grey N, Chilcott RP (2021) SARS-CoV-2: characterisation and mitigation of risks associated with aerosol generating procedures in dental practices. Br Dent J 7:1–7. https://doi.org/10. 1038/s41415-020-2504-8 52. Shahdad S, Hindocha A, Patel T, Cagney N, Mueller JD, Koched A, Seoudi N, Morgan C, Fleming PS, Din AR (2021) Fallow time determination in dentistry using aerosol measurement in mechanically and non-mechanically ventilated environments. Br Dent J 24:1–8. https://doi.org/10.1038/ s41415-021-3369-1 53. Beltrán EO, Newton JT, Avila V, Pitts NB, Castellanos JE, Tenuta LMA, Martignon S (2022) Dentists’ perceptions of personal infection control measurements in response to COVID-19. JDR Clin Trans Res 7:23800844221123750. https://doi.org/10.1177/23800 844221123751 |
| dc.relation.citationendpage.spa.fl_str_mv |
3231 |
| dc.relation.citationstartpage.spa.fl_str_mv |
3221 |
| dc.relation.citationvolume.spa.fl_str_mv |
27 |
| dc.rights.eng.fl_str_mv |
© The Author(s) 2023 |
| dc.rights.license.spa.fl_str_mv |
Atribución 4.0 Internacional (CC BY 4.0) |
| dc.rights.uri.spa.fl_str_mv |
https://creativecommons.org/licenses/by/4.0/ |
| dc.rights.accessrights.spa.fl_str_mv |
info:eu-repo/semantics/embargoedAccess |
| dc.rights.coar.spa.fl_str_mv |
http://purl.org/coar/access_right/c_f1cf |
| rights_invalid_str_mv |
Atribución 4.0 Internacional (CC BY 4.0) © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/ http://purl.org/coar/access_right/c_f1cf |
| eu_rights_str_mv |
embargoedAccess |
| dc.format.extent.spa.fl_str_mv |
11 páginas |
| dc.format.mimetype.spa.fl_str_mv |
application/pdf |
| dc.publisher.spa.fl_str_mv |
Springer Verlag |
| dc.publisher.place.spa.fl_str_mv |
Germany |
| dc.source.spa.fl_str_mv |
https://link.springer.com/article/10.1007/s00784-023-04937-z |
| institution |
Corporación Universidad de la Costa |
| bitstream.url.fl_str_mv |
https://repositorio.cuc.edu.co/bitstreams/17dd6e0c-ce0f-4450-8053-a6b0159981d0/download https://repositorio.cuc.edu.co/bitstreams/1c2abef4-aa42-4d53-957d-a00fbc7e7f26/download https://repositorio.cuc.edu.co/bitstreams/6ae56f4b-4564-4aa9-bf86-f031fd472459/download https://repositorio.cuc.edu.co/bitstreams/9bb20812-8ad3-435d-b444-88ec123c9438/download |
| bitstream.checksum.fl_str_mv |
75ba80ddd841392e1d0cb4f892d46032 2f9959eaf5b71fae44bbf9ec84150c7a e636a3ad8f00e8714d5138e2d4c39c02 a517cdda3d6c6f2cc60c17a367f75778 |
| bitstream.checksumAlgorithm.fl_str_mv |
MD5 MD5 MD5 MD5 |
| repository.name.fl_str_mv |
Repositorio de la Universidad de la Costa CUC |
| repository.mail.fl_str_mv |
repdigital@cuc.edu.co |
| _version_ |
1811760701404872704 |
| spelling |
Atribución 4.0 Internacional (CC BY 4.0)© The Author(s) 2023https://creativecommons.org/licenses/by/4.0/info:eu-repo/semantics/embargoedAccesshttp://purl.org/coar/access_right/c_f1cfBeltrán, Edgar O.Castellanos, Jaime E.Corredor, Zayda L.Morgado, WendyZarta, Olga L.Cortés, AndreaAvila, VivianaMartignon, Stefania2023-09-11T18:57:50Z20242023-09-11T18:57:50Z2023Beltrán, E.O., Castellanos, J.E., Corredor, Z.L. et al. Tracing ΦX174 bacteriophage spreading during aerosol-generating procedures in a dental clinic. Clin Oral Invest 27, 3221–3231 (2023). https://doi.org/10.1007/s00784-023-04937-z1432-6981https://hdl.handle.net/11323/1046910.1007/s00784-023-04937-z1436-3771Corporación Universidad de la CostaREDICUC - Repositorio CUChttps://repositorio.cuc.edu.co/Objective The aim of this study was to test the plausibility of using the ΦX174 bacteriophage as a tracer of viral aerosols spreading in a dental aerosol-generating procedure (AGP) model. Methods ΦX174 bacteriophage (~ 108 plaque-forming units (PFU)/mL) was added into instrument irrigation reservoirs and aerosolized during class-IV cavity preparations followed by composite fllings on natural upper-anterior teeth (n=3) in a phantom head. Droplets/aerosols were sampled through a passive approach that consisted of Escherichia coli strain C600 cultures immersed in a LB top agar layer in Petri dishes (PDs) in a double-layer technique. In addition, an active approach consisted of E coli C600 on PDs sets mounted in a six-stage cascade Andersen impactor (AI) (simulating human inhalation). The AI was located at 30 cm from the mannequin during AGP and afterwards at 1.5 m. After collection PDs were incubated overnight (18 h at 37 °C) and bacterial lysis was quantifed. Results The passive approach disclosed PFUs mainly concentrated over the dental practitioner, on the mannequin’s chest and shoulder and up to 90 cm apart, facing the opposite side of the AGP’s source (around the spittoon). The maximum aerosol spreading distance was 1.5 m in front of the mannequin’s mouth. The active approach disclosed collection of PFUs corresponding to stages (and aerodynamic diameters) 5 (1.1–2.1 µm) and 6 (0.65–1.1 µm), mimicking access to the lower respiratory airways. Conclusion The ΦX174 bacteriophage can be used as a traceable viral surrogate in simulated studies contributing to understand dental bioaerosol’s behavior, its spreading, and its potential threat for upper and lower respiratory tract. Clinical relevance The probability to fnd infectious virus during AGPs is high. This suggests the need to continue characterizing the spreading viral agents in diferent clinical settings through combination of passive and active approaches. In addition, subsequent identifcation and implementation of virus-related mitigation strategies is relevant to avoid occupational virus infections.11 páginasapplication/pdfengSpringer VerlagGermanyhttps://link.springer.com/article/10.1007/s00784-023-04937-zTracing ΦX174 bacteriophage spreading during aerosol‑generating procedures in a dental clinicArtículo de revistahttp://purl.org/coar/resource_type/c_2df8fbb1Textinfo:eu-repo/semantics/articlehttp://purl.org/redcol/resource_type/ARTinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/version/c_970fb48d4fbd8a85Clinical Oral Investigations1. Ren Y, Feng C, Rasubala L, Malmstrom H, Eliav E (2020) Risk for dental healthcare professionals during the COVID-19 global pandemic: an evidence-based assessment. J Dent 101:103434. https://doi.org/10.1016/j.jdent.2020.1034342. Odeh ND, Babkair H, Abu-Hammad S, Borzangy S, AbuHammad A, Abu-Hammad O (2020) COVID-19: present and future challenges for dental practice. Int J Environ Res Public Health 17(9):3151. https://doi.org/10.3390/ijerph170931513. World Health Organization (2020) Transmission of SARS-CoV-2: implications for infection prevention precautions. Available at: https://www.who.int/emergencies/diseases/novel-coronavirus2019?gclid=Cj0KCQjw0YD4BRD2ARIsAHwmKVmNyTZntqH Y8mnWiIWPlKpj41bEGY-UjmYkbJ-6mjfBTYlMQrrQLV8aAtD REALw_wcB. Accessed June 20234. Meng L, Hua F, Bian Z (2020) Coronavirus disease 2019 (COVID-19): emerging and future challenges for dental and oral medicine. J Dent Res 99(5):481–487. https://doi.org/10.1177/ 00220345209142465. Allison JR, Dowson C, Jakubovics NS, Nile C, Durham J, Holliday R (2022) Waterline disinfectants reduce dental bioaerosols: a multitracer validation. J Dent Res 101(10):1198–1204. https:// doi.org/10.1177/002203452210935226. Bourouiba L (2022) Turbulent gas clouds and respiratory pathogen emissions: potential implications for reducing transmission of COVID-19. JAMA 323(18):1837–1838. https://doi.org/10.1001/ jama.2020.47567. Zemouri C, Volgenant CMC, Buijs MJ, Crielaard W, Rosema NAM, Brandt BW, Laheij AMGA, De Soet JJ (2020) Dental aerosols: microbial composition and spatial distribution. J Oral Microbiol 12(1):1762040. https://doi.org/10.1080/20002297. 2020.17620408. Kumbargere Nagraj S, Eachempati P, Paisi M, Nasser M, Sivaramakrishnan G, Verbeek JH (2020) Interventions to reduce contaminated aerosols produced during dental procedures for preventing infectious diseases. Cochrane Database Syst Rev 10(10):CD013686. https://doi.org/10.1002/14651 858.CD013686.pub29. Allison JR, Currie CC, Edwards DC, Bowes C, Coulter J, Pickering K, Kozhevnikova E, Durham J, Nile CJ, Jakubovics N, Rostami N, Holliday R (2021) Evaluating aerosol and splatter following dental procedures: addressing new challenges for oral health care and rehabilitation. J Oral Rehabil 48(1):61–72. https://doi.org/10.1111/joor.1309810. Holliday R, Allison JR, Currie CC, Edwards DC, Bowes C, Pickering K, Reay S, Durham J, Lumb J, Rostami N, Coulter J, Nile C, Jakubovics N (2021) Evaluating contaminated dental aerosol and splatter in an open plan clinic environment: implications for the COVID-19 pandemic. J Dent 105:103565. https:// doi.org/10.1016/j.jdent.2020.10356511. Llandro H, Allison JR, Currie CC, Edwards DC, Bowes C, Durham J, Jakubovics N, Rostami N, Holliday R (2021) Evaluating splatter and settled aerosol during orthodontic debonding: implications for the COVID-19 pandemic. Br Dent J 8:1–7. https://doi.org/10.1038/s41415-020-2503-912. Din AR, Hindocha A, Patel T, Sudarshan S, Cagney N, Koched A, Mueller JD, Seoudi N, Morgan C, Shahdad S, Fleming PS (2020). Quantitative analysis of particulate matter release during orthodontic procedures: a pilot study. Br Dent J 12:1–7. https://doi.org/10.1038/s41415-020-2280-513. Timmerman MF, Menso L, Steinfort J, van Winkelhof AJ, van der Weijden GA (2004) Atmospheric contamination during ultrasonic scaling. J Clin Periodontol 31(6):458–462. https:// doi.org/10.1111/j.1600-051X.2004.00511.x14. Rautemaa R, Nordberg A, Wuolijoki-Saaristo K, Meurman JH (2006) Bacterial aerosols in dental practice – a potential hospital infection problem? J Hosp Infect 64(1):76–81. https://doi. org/10.1016/j.jhin.2006.04.01115. Holloman JL, Mauriello SM, Pimenta L, Arnold RR (2015) Comparison of suction device with saliva ejector for aerosol and spatter reduction during ultrasonic scaling. J Am Dent Assoc 146(1):27–33. https://doi.org/10.1016/j.adaj.2014.10.00116. Bennett AM, Fulford MR, Walker JT, Bradshaw DJ, Martin MV, Marsh PD (2020) Microbial aerosols in general dental practice. Br Dent J 189(12):664–667. https://doi.org/10.1038/sj.bdj.480085917. Dutil S, Meriaux A, de Latremoille MC, Lazure L, Barbeau J, Duchaine C (2009) Measurement of airborne bacteria and endotoxin generated during dental cleaning. J Occup Environ Hyg 6(2):121–130. https://doi.org/10.1080/1545962080263395718. King MD, McFarland AR (2012) Use of an Andersen bioaerosol sampler to simultaneously provide culturable particle and culturable organism size distributions. Aerosol Sci Technol 46(8):852–861. https://doi.org/10.1080/02786826.2012.66950719. Gendron L, Verreault D, Veillette M, Moineau S, Duchaine C (2010) Evaluation of flters for the sampling and quantifcation of RNA phage aerosols. Aerosol Sci Technol 44:893–901. https://doi.org/10.1080/02786826.2010.50135120. Krupovic M, Bamford DH (2008) Virus evolution: how far does the double beta-barrel viral lineage extend? Nat Rev Microbiol 6(12):941–948. https://doi.org/10.1038/nrmicro203321. Verreault D, Moineau S, Duchaine C (2008) Methods for sampling of airborne viruses. Microbiol Mol Biol Rev 72(3):413–444. https://doi.org/10.1128/MMBR.00002-0822. Verreault D, Rousseau GM, Gendron L, Masse D, Moineau S, Duchaine C (2010) Comparison of polycarbonate and polytetrafuoroethylene flters for sampling of airborne bacteriophages. Aerosol Sci Technol 44:197–201. https://doi.org/10.1080/02786 82090351889923. Turgeon N, Toulouse MJ, Martel B, Moineau S, Duchaine C (2014) Comparison of fve bacteriophages as models for viral aerosol studies. Appl Environ Microbiol 80(14):4242–4250. https://doi.org/10.1128/AEM.00767-1424. Appleyard RK (1954) Segregation of new lysogenic types during growth of a doubly lysogenic strain derived from Escherichia coli K12. Genetics 39(4):440–452. https://doi.org/10.1093/genetics/39.4.44025. Kropinski AM, Mazzocco A, Waddell TE, Lingohr E, Johnson RP (2009) Enumeration of bacteriophages by double agar overlay plaque assay. Methods Mol Biol 501:69–76. https://doi.org/10. 1007/978-1-60327-164-6_726. Vernon JJ, Black EVI, Dennis T, Devine DA, Fletcher L, Wood DJ, Nattress BR (2021) Dental mitigation strategies to reduce aerosolization of SARS-CoV-2. J Dent Res 100(13):1461–1467. https://doi.org/10.1177/0022034521103288527. Rheinbaben F, Schunemann S, Gross T, Wolf MH (2000) Transmission of viruses via contact in a household setting: experiments using bacteriophage straight phiX174 as a model virus. J Hosp Infect 46:61–6628. Mik G, de Groot I, Gerbrandy JL (1977) Survival of aerosolized bacteriophage phiX174 in air containing ozone-olefn mixtures. J Hyg (London) 78:189–19829. Dbouk T, Drikakis D (2020) On coughing and airborne droplet transmission to humans. Phys Fluids 32(5):053310. https://doi. org/10.1063/5.001196030. Chen L, Zhao J, Peng J, Li X, Deng X, Geng Z, Shen Z, Guo F, Zhang Q, Jin Y, Wang L, Wang S (2020) Detection of SARS-CoV-2 in saliva and characterization of oral symptoms in COVID-19 patients. Cell Prolif 53(12):e12923. https://doi.org/10.1111/cpr.1292331. Teo AKJ, Choudhury Y, Tan IB, Cher CY, Chew SH, Wan ZY, Cheng LTE, Oon LLE, Tan MH, Chan KS, Hsu LY (2021) Saliva is more sensitive than nasopharyngeal or nasal swabs for diagnosis of asymptomatic and mild COVID-19 infection. Sci Rep 11(1):3134. https://doi.org/10.1038/s41598-021-82787-z. (Erratum.In:SciRep.2021Jun9;11(1):12538)32. Leung NHL (2021) Transmissibility and transmission of respiratory viruses. Nat Rev Microbiol 19(8):528–545. https://doi.org/ 10.1038/s41579-021-00535-633. Gupta G, Mitra D, Ashok KP, Gupta A, Soni S, Ahmed S, Arya A (2014) Efcacy of preprocedural mouth rinsing in reducing aerosol contamination produced by ultrasonic scaler: a pilot study. J Periodontol 85(4):562–568. https://doi.org/10.1902/jop.2013.12061634. To KK, Tsang OT, Yip CC, Chan KH, Wu TC, Chan JM, Leung WS, Chik TS, Choi CY, Kandamby DH, Lung DC, Tam AR, Poon RW, Fung AY, Hung IF, Cheng VC, Chan JF, Yuen KY (2020) Consistent detection of 2019 novel coronavirus in saliva. Clin Infect Dis 71(15):841–843. https://doi.org/10.1093/cid/ciaa14935. Paton S, Clark S, Spencer A, Garratt I, Dinesh I, Thompson KA, Bennett A, Pottage T (2022) Characterisation of particle size and viability of SARS-CoV-2 aerosols from a range of nebuliser types using a novel sampling technique. Viruses 19;14(3):639. https:// doi.org/10.3390/v1403063936. Fröhlich-Nowoisky J, Kampf CJ, Weber B, Hufman JA, Pöhlker C, Andreae MO, Lang-Yona N, Burrows SM, Gunthe SS, Elbert W, Su H, Hoor P, Thines E, Hofmann T, Després VR, Pöschl U (2016) Bioaerosols in the Earth system: climate, health, and ecosystem interactions. Atmos Res 182:346–376, ISSN 0169–8095. https://doi.org/10.1016/j.atmosres.2016.07.01837. Morgado-Gamero WB, Mendoza Hernandez M, Castillo Ramirez M, Medina-Altahona J, De La Hoz S, Posso Mendoza H, Parody A, Teixeira EC, Agudelo-Castañeda DM (2019) Antibiotic resistance of airborne viable bacteria and size distribution in neonatal intensive care units. Int J Environ Res Public Health 10;16(18):3340. https://doi.org/10. 3390/ijerph1618334038. Morgado-Gamero WB, Parody A, Medina J, Rodriguez-Villamizar LA, Agudelo-Castañeda D (2021) Multi-antibiotic resistant bacteria in landfll bioaerosols: environmental conditions and biological risk assessment. Environ Pollut 1 290:118037. https://doi.org/10. 1016/j.envpol.2021.11803739. Broussard IM, Kahwaji CI. Universal Precautions. [Updated 2022 Sep 1]. In: StatPearls [Internet]. Treasure Island (FL): StatPearls Publishing; 2022 Jan-. Available from: https://www.ncbi.nlm.nih. gov/books/NBK470223/40. ASHRAE (2020a) COVID-19 (CORONAVIRUS) Preparedness resources. American Society of Heating, Ventilating, and Air- Conditioning Engineers. Available from: https://www.ashrae.org/fle% 20library/technical%20resources/covid-19/in-room-air-cleaner-guida nce-for-reducing-covid-19-in-air-in-your-space-or-room.pdf41. ASHRAE (2020b) Position Document on Airborne Infectious Diseases, Approved by the Board of Directors, January 19, 2014. Reafrmed by the Technology Council, February 5, 2020. Atlanta, Georgia42. Occupational Safety and Health Administration (2011) Dentistry workers and employers. Available at https://www.osha.gov/SLTC/ covid-19/dentistry.html. Accessed 28 May 202043. Gund M, Isack J, Hannig M, Thieme-Rufng S, Grtner B, Boros G et al (2021) Contamination of surgical mask during aerosolproducing dental treatments. Clin Oral Investig 25:3173–3180. https://doi.org/10.1007/s00784-020-03645-244. Balanta-Melo J, Gutiérrez A, Sinisterra G, Díaz-Posso MdM, Gallego D, Villavicencio J, Contreras A (2020) Rubber Dam isolation and high-volume suction reduce ultrafne dental aerosol particles: an experiment in a simulated patient. Appl Sci 10(18):6345. https://doi.org/10.3390/app1018634545. Morawska L, Tang JW, Bahnfleth W, Bluyssen PM, Boerstra A, Buonanno G, Cao J, Dancer S, Floto A, Franchimon F, Haworth C, Hogeling J, Isaxon C, Jimenez JL, Kurnitski J, Li Y, Loomans M, Marks G, Marr LC, Mazzarella L, Melikov AK, Miller S, Milton DK, Nazaroff W, Nielsen PV, Noakes C, Peccia J, Querol X, Sekhar C, Seppänen O, Tanabe SI, Tellier R, Tham KW, Wargocki P, Wierzbicka A, Yao M (2020) How can airborne transmission of COVID-19 indoors be minimised? Environ Int 142:105832. https://doi.org/10.1016/j.envint.2020. 10583246. Bai Y, Yao L, Wei T, Tian F, Jin DY, Chen L, Wang M (2020) Presumed asymptomatic carrier transmission of COVID-19. JAMA 323(14):1406–1407. https://doi.org/10.1001/jama.2020. 256547. Ren YF, Huang Q, Marzouk T, Richard R, Pembroke K, Martone P, Venner T, Malmstrom H, Eliav E (2020) Efects of mechanical ventilation and portable air cleaner on aerosol removal from dental treatment rooms. J Dent 105:103576. https://doi.org/10. 1016/j.jdent.2020.10357648. Hiroshi N, Hitoshi O, Tetsuro K, Tomoya S, Tomimasa S, Tamano M, Takaji W, MHLW COVID-19 Response Team, Motoi Suzuki (2020) Closed environments facilitate secondary transmission of coronavirus disease 2019 (COVID-19). medRxiv 2020.02.28.20029272. https:// doi.org/10.1101/2020.02.28.2002927249. Godwin CC, Batterman SA, Sahni SP, Peng CY (2003) Indoor environment quality in dental clinics: potential concerns from particulate matter. Am J Dent 16(4):260–26650. Robertson C, Clarkson JE, Aceves-Martins M, Ramsay CR, Richards D, Colloc T, CoDER Working Group (2021) A review of aerosol generation mitigation in international dental guidance. Int Dent J 72(2):203–210. https://doi.org/10.1016/j. identj.2021.04.00251. Ehtezazi T, Evans DG, Jenkinson ID, Evans PA, Vadgama VJ, Vadgama J, Jarad F, Grey N, Chilcott RP (2021) SARS-CoV-2: characterisation and mitigation of risks associated with aerosol generating procedures in dental practices. Br Dent J 7:1–7. https://doi.org/10. 1038/s41415-020-2504-852. Shahdad S, Hindocha A, Patel T, Cagney N, Mueller JD, Koched A, Seoudi N, Morgan C, Fleming PS, Din AR (2021) Fallow time determination in dentistry using aerosol measurement in mechanically and non-mechanically ventilated environments. Br Dent J 24:1–8. https://doi.org/10.1038/ s41415-021-3369-153. Beltrán EO, Newton JT, Avila V, Pitts NB, Castellanos JE, Tenuta LMA, Martignon S (2022) Dentists’ perceptions of personal infection control measurements in response to COVID-19. JDR Clin Trans Res 7:23800844221123750. https://doi.org/10.1177/23800 8442211237513231322127BacteriophageAerosolAerosol-generating procedureDental settingVirus laden aerosolsAirborne transmissionPublicationORIGINALTracing ΦX174 bacteriophage spreading during aerosol‑generating.pdfTracing ΦX174 bacteriophage spreading during aerosol‑generating.pdfArtículoapplication/pdf1725458https://repositorio.cuc.edu.co/bitstreams/17dd6e0c-ce0f-4450-8053-a6b0159981d0/download75ba80ddd841392e1d0cb4f892d46032MD51LICENSElicense.txtlicense.txttext/plain; charset=utf-814828https://repositorio.cuc.edu.co/bitstreams/1c2abef4-aa42-4d53-957d-a00fbc7e7f26/download2f9959eaf5b71fae44bbf9ec84150c7aMD52TEXTTracing ΦX174 bacteriophage spreading during aerosol‑generating.pdf.txtTracing ΦX174 bacteriophage spreading during aerosol‑generating.pdf.txtExtracted texttext/plain49620https://repositorio.cuc.edu.co/bitstreams/6ae56f4b-4564-4aa9-bf86-f031fd472459/downloade636a3ad8f00e8714d5138e2d4c39c02MD53THUMBNAILTracing ΦX174 bacteriophage spreading during aerosol‑generating.pdf.jpgTracing ΦX174 bacteriophage spreading during aerosol‑generating.pdf.jpgGenerated Thumbnailimage/jpeg15279https://repositorio.cuc.edu.co/bitstreams/9bb20812-8ad3-435d-b444-88ec123c9438/downloada517cdda3d6c6f2cc60c17a367f75778MD5411323/10469oai:repositorio.cuc.edu.co:11323/104692024-09-17 10:18:05.095https://creativecommons.org/licenses/by/4.0/© The Author(s) 2023open.accesshttps://repositorio.cuc.edu.coRepositorio de la Universidad de la Costa CUCrepdigital@cuc.edu.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 |