Project EXPOsE - Project nº 23222 (02/SAICT/2016)Main project: EXPOsE – Establishing protocols to assess occupational exposure to microbiota in clinical settings. Our study: Correlational analysis of the relationship between variables - Does the air quality of healthcare facilities influences biological responses? Healthcare facilities display an important role in the transmission of airborne infections and risk assessment to promote patients and professionals safety.info:eu-repo/semantics/publishedVersio

Caetano, Liliana Aranha

Carolino, Elisabete

Costa, Natália

Oliveira, Cátia

English

Repositório Científico do Instituto Politécnico de Lisboa

Indoor Air Quality in Portuguese Healthcare FacilitiesA Correlational Study on Environmental Parameters andCytotoxic and Pro-inflammatory Effects Instituto Politécnico de Lisboa – Escola Superior de Tecnologia da Saúde de Lisboa I Webinar de Investigação Aplicada em FarmáciaAutores: Natália Costa e Cátia OliveiraProf.ª Liliana Aranha Caetano e Prof.ª Elisabete CarolinoOur StudyMain project: EXPOsE – Establishing protocols to assess occupational exposure to microbiota in clinical settings; Financed by FCT – Fundação para a Ciência e Tecnologia (02/SAICT/2016 –Project n-º 23222) and by the Polish National agency of Academic Exchange (PPN/BIL/2018/1/00231)2Our StudyCorrelational analysis of the relationship between variables“Does the air quality of healthcare facilities influences biological responses?”“Indoor Air Quality”, “Bioaerosols”, “Nosocomial Infections” and “Assessment Methods of Environmental Contamination”Healthcare facilities display an important role in the transmission of airborne infections Risk assessment to promote patients and professionals safety3Indoor Air QualityEnvironmental Conditions Concentration of PollutantsNegative health impactAssessment of environmental parametersAssessment of contamination41. Kubba S. Butterworth-Heinemann; 2017 2. Soreanu G. Elsevier; 20161-2Bioaerosols and Nosocomial InfectionsBioaerosols are particles with biological originsInfections, irritations and allergiesNosocomial infections⚠️Methods for controlling airborne pathogens 0,02µm 1000µm4µm 10µmBioaerosols Size RangeRespirable Particles Size Range52-92. Sivagnanasundaram P. BioMed Research International; 2019; 3. Qudiesat K, African Journal of Microbiology Research. 2009; 4. Eduard W. SJWEH; 2009; 5. Haig CW. J Hosp Infect; 2016; 6. Douwes J. Ann Occup Hy; 2003; 7. Nazaroff WW. Indoor Air; 2016; 8. Sykes P. Waste Manag.; 2011; 9. Wéry N. Front Cell Infect Microbiol; 2014; 10. Brown JS. Part Fibre Toxicol; 20135, 10Methods of Assessing Environmental ContaminationSampling Methods 10, 11Active MethodsAutomatic Air SamplersImpactionPassive MethodsSurface SwabbingSettle DustCombination of Both610. Viegas C; Int J Environ Health Res. 2019; 11. Napoli C; BMC Public Health. 2012Biological Response to BioaerosolsMeasured using:♙Cytotoxic effects on THP-1 cells 12, 13; ♙Cytotoxic effects on Calu-3 cells 14;♙Pro-inflammatory effects of cytokines IL-1β and TNF-α (from THP-1 cells) 12, 13712. Viegas S; Toxics. 2017; 13. Wang Z; Eur Respir J. 1997; 14. Matsuwaki Y; Int Arch Allergy Immunol. 2012ObjectivesBiological ResponseMicrobiological contamination (including metabolites – e.g.: mycotoxins)Non-biological parameters (e.g.: Temperature)8Materials and MethodsEnvironmental Sampling10 PHCUs1 Central HospitalCulture-base MethodsMolecular MethodsDetection of Microorganisms and MetabolitesIn vitro Biological Response AssaysStatistical Analysis9Statistical Analysis10Spearman Correlation TestsLinear Regression Analysis -ANOVAChi-Square + Kruskal-Wallis TestsIdentify Statistically Significant Differences Identify Statistically Significant CorrelationsIdentify Prediction Factors of the Biological ResponseMain Results11Fungal contamination (culture-based methods)THP-1 Cell viabilityFungal Biomass Calu-3 Cell viability Strong and inversecorrelations showed bySpearman Correlation Testsfor PHCUsPrevious study with THP-1 cells’ response to air quality→ pro-inflammatory effects (of cytokines TNF-α and IL-1β) +cytotoxic effect on all air samples collected 1212. Viegas S; Toxics. 2017Main Results12y = -0,025x + 145,75145,45145,5145,55145,6145,65145,7145,75145,80 2 4 6 8 10 12IL-1β ' Secretion on Air Samples ρg/mLFungal Contamination of Air Samples in MEA CFU/m2 y = -0,001x + 75,07275,06675,06775,06875,06975,0775,07175,07275,0730 1 2 3 4 5 6IL-1β's Secretion on Air Samples ρg/mLFungal Contamination of Swabs Samples in MEA CFU/m2Figure 1 - Linear Regression Analysis (ANOVA Test), model 1 Figure 2 - Linear Regression Analysis (ANOVA Test), model 4Fungal contamination decreases IL-1β’ secretionMain Results13Fungal contamination with Gram (-) bacteria was mainly associated with decreased cell viabilityin THP-1 cells in air samplesVariables (air contamination CFU/m2)Influence over dependent variable (THP-1 cells’ relative viability %)VRBA (Gram (-) bacterial contamination)-0.272MEA (fungal contamination) -0.18DG18 (fungal contamination) 0.06Variables (air contamination CFU/m2)Influence over dependent variable (Calu-3 cells’ relative viability %)Total TSA (bacterial contamination)-0.065VRBA (Gram (-) bacterial contamination)-4.183Figure 3 - Linear Regression Analysis (ANOVA Test), model 2Figure 4 - Linear Regression Analysis (ANOVA Test), model 3Gram (-) bacteria combined with total bacterial contamination (on TSA), on air samples,decreases cell viability of Calu-3Main Results14PHCU 5, which had the highest count of bacterial contamination (on TSA) also had the lowest count of Calu-3 and THP-1 viabilities.45,2138,8824,7527,3660,7532,8155,5025,9443,5044,0033,1436,1923,8637,8326,9442,3852,5046,6964,3626,19P H C U  1 P H C U  2 P H C U  3 P H C U  4 P H C U  5 P H C U  6 P H C U  7 P H C U  8 P H C U  9 P H C U  1 0BACTERIAL CONTAMINATION (MEAN RANK CFU/M2)TSA Air Samples TSA Swabs Samples12,4013,5018,5014,003,7526,25 29,5026,5035,0026,0029,8022,7519,2514,005,5013,2524,2516,5030,0025,75P H C U  1 P H C U  2 P H C U  3 P H C U  4 P H C U  5 P H C U  6 P H C U  7 P H C U  8 P H C U  9 P H C U  1 0CELLS' VIABILITY (MEAN RANK %)Calu-3 cells THP-1Figure 6 - Mean Ranks (results of Kruskal-Wallis test) to compare biological responses of both cell lines between PHCUsFigure 5 - Mean Ranks (results of Kruskal-Wallis test) to compare bacterial counts between PHCUsMain Results1577,7160,3854,4455,8610,5020,63 26,0518,7537,2158,3865,2957,6348,1137,5728,7541,0026,9116,0039,1456,3811,715,0041,2239,5734,2569,1361,6448,5058,2931,2540,5739,1361,7841,8658,5032,0014,3610,6363,8657,6319,43 24,6365,7819,5751,0057,5037,4515,1442,1462,1335,7157,5054,5629,8623,6341,2540,0916,8636,2962,2565,1449,0060,2251,7139,6325,7521,008,8848,8650,25P H C U  1 P H C U  2 P H C U  3 P H C U  4 P H C U  5 P H C U  6 P H C U  7 P H C U  8 P H C U  9 P H C U  1 0MEAN RANKCO Level (mg/m3) CO2 Level (mg/m3) Temperature (ºC) Relative Humidity (%)PM 2.5 (μg/m3) PM 10 (μg/m3) VOCs (μg/m3)Figure 8 - Mean Ranks (results of Kruskal-Wallis test) to compare environmental parameters between PHCUsMain Results1630,0742,6925,8343,7947,9447,6329,1455,7558,5036,1936,5028,63 35,5038,6435,8849,4429,1459,7542,2158,757,7118,0053,4443,2139,0626,0658,6857,4446,2148,7519,5718,0055,3942,7922,1934,8160,7750,8841,5752,44P H C U  1 P H C U  2 P H C U  3 P H C U  4 P H C U  5 P H C U  6 P H C U  7 P H C U  8 P H C U  9 P H C U  1 0FUNGAL CONTAMINATION (MEAN RANK CFU/M2)MEA Swabs Samples DG18 Swabs Samples MEA Air Samples DG18 Air Samples45,2138,8824,7527,3660,7532,8155,5025,9443,5044,0033,1436,1923,8637,8326,9442,38 52,5046,6964,3626,19P H C U  1 P H C U  2 P H C U  3 P H C U  4 P H C U  5 P H C U  6 P H C U  7 P H C U  8 P H C U  9 P H C U  1 0BACTERIAL CONTAMINATION (MEAN RANK CFU/M2)TSA Air Samples TSA Swabs SamplesFigure 9 Mean Ranks (results of Kruskal-Wallis test) to compare fungal counts between PHCUsFigure 10 - Mean Ranks (results of Kruskal-Wallis test) to compare bacterial counts between PHCUsMain Results1712,4013,5018,5014,003,7526,25 29,5026,5035,0026,0029,8022,7519,2514,005,5013,2524,2516,5030,0025,75P H C U  1 P H C U  2 P H C U  3 P H C U  4 P H C U  5 P H C U  6 P H C U  7 P H C U  8 P H C U  9 P H C U  1 0CELLS' VIABILITY (MEAN RANK %)Calu-3 cells THP-1Figure 11 - Mean Ranks (results of Kruskal-Wallis test) to compare biological responses of both cell lines between PHCUsHigher levels of carbon monoxide and dioxide  may reduce viable fungal and bacterial air contaminants and allow higher viability of THP-1 and Calu-3 cells. Main Results1840,0040,0040,0040,00 45,0645,0640,0040,0040,0040,0038,0038,00 42,5043,79 48,1348,1338,0038,0038,0038,0040,5040,5040,5040,50 45,5640,5040,5040,5040,5040,5040,5040,5040,5040,50 45,5640,5040,5040,5040,5040,5037,0037,00 41,5042,79 47,1357,2537,0037,0037,0037,0040,5040,5040,5040,5040,50 45,5640,5040,5040,5040,5041,0041,0041,0041,0041,0041,0041,0041,0041,0041,0041,0041,0041,0041,0041,0041,0041,0041,0041,0041,00P H C U  1 P H C U  2 P H C U  3 P H C U  4 P H C U  5 P H C U  6 P H C U  7 P H C U  8 P H C U  9 P H C U  1 0MYCOTOXIN LOAD (ΜG/KG)Presence of Fumonisin B1 Presence of Fumonisin B2 Presence of Fumonisin B3Presence of Roquefortine C Presence of Ochratoxin A Presence of Ochratoxin BPresence of Mycophenolic Acid Presence of Sterigmatocystin12,4013,50 18,5014,003,7526,2529,5026,5035,0026,0029,8022,7519,2514,005,5013,2524,2516,5030,0025,75P H C U  1 P H C U  2 P H C U  3 P H C U  4 P H C U  5 P H C U  6 P H C U  7 P H C U  8 P H C U  9 P H C U  1 0CELLS' VIABILITY (MEAN RANK %)Calu-3 cells THP-120,2030,7529,2523,3823,8832,0016,253,0013,25 18,2533,2026,0034,2531,0016,7526,009,505,38 10,3814,50P H C U  1 P H C U  2 P H C U  3 P H C U  4 P H C U  5 P H C U  6 P H C U  7 P H C U  8 P H C U  9 P H C U  1 0CYTOKINES' SECRETION (MEAN RANK ΡG/ML)IL-1β cytokines TNF-α cytokinesOchratoxin A has been associated with decrease in metabolic activity, cellproliferation, differentiation, phagocytic behavior and other effects over THP-1 cells and Fumonisin 1 with decreased cell viability also in THP-1 cells 17, 18Figure 13 - Mean Ranks (results of Kruskal-Wallis test) to compare mycotoxin counts between PHCUs8Figure 14 - Mean Ranks (results of Kruskal-Wallis test) to compare biological responses of both cell lines between PHCUsFigure 15 - Mean Ranks (results of Kruskal-Wallis test) to compare biological responses of both cytokines between PHCUs17. Müller G; Toxicology. 2003, 18. Smith M-C; Cell Biol Toxicol. 2018Main Results19FumonisinB1RoquefortineCFumonisin B3Ochratoxin BTrichothecene mycotoxins and aflatoxins can beefficiently inactivated with common cleaningproducts, such as sodium hypochlorite 19, 20Strong and direct correlations showed by Spearman Correlation Tests for PHCUs19. Wilson, SC; J Occup Environ Hyg. 2004;20. Yang, CY; Appl Microbiol. 1972;Main Results2052,6346,13 51,6852,7919,7054,6734,2030,00D G 1 8  A I R  S A M P L E SM E A  A I R  S A M P L E SD G 1 8  S W A B  S A M P L E S  M E A  S W A B S  S A M P L E SFUNGAL CONTAMINATION (MEAN RANK CFU/M2)PHCUs Hospital48,3949,9353,4949,4442,0742,0010,7045,50T S A  A I R  S A M P L E SR B  A I R  S A M P L E ST S A  S W A B S  S A M P L E SR B  S W A B S  S A M P L E SBACTERIAL CONTAMINATION (MEAN RANK CFU/M2)PHCUs Hospital53,2855,3452,5653,4442,0449,7343,0517,1317,9318,1315,0788,5036,5078,67MEAN RANKPHCUs HospitalFigure 16 - Mean Ranks (results of Kruskal-Wallis test) to compare bacterial counts between all PHCUs and the HospitalFigure 17 - Mean Ranks (results of Kruskal-Wallis test) to compare fungal counts between all PHCUs and the HospitalFigure 18 - Mean Ranks (results of Kruskal-Wallis test) to compare non-biological parameters between all PHCUs and the HospitalLower contamination offungi and bacteria + highercarbon dioxide and VOCsMain Findings♙Indoor air quality parameters associated with microbiological and non-biological contaminants mayinfluence the biological response in Calu-3 and TPH-1 cells;♙Fungal and bacterial (namely gram (-) specimens) load, carbon monoxide levels, temperature and relativehumidity proved to be statistically significant influencers of the biological response;♙Correlations between the presence of different mycotoxins in air samples may suggest improper practicesand/or inefficient cleaning and disinfecting methods.21Thank youIndoor Air Quality in Portuguese Healthcare FacilitiesA Correlational Study on Environmental Parameters andCytotoxic and Pro-inflammatory Effects 

Indoor air quality in Portuguese healthcare facilities: a correlational study on environmental parameters and cytotoxic and pro-inflammatory effects

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Indoor air quality in Portuguese healthcare facilities: a correlational study on environmental parameters and cytotoxic and pro-inflammatory effects

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