key: cord-0972103-5zr5l1xl authors: Silva, S. J.; Nascimento, J.; Reis, W.; Silva, C.; Silva, P.; Mendes, R.; Mendonca, A.; Santos, B.; Magalhaes, J.; Kohl, A.; Pena, L. e. title: Widespread Contamination of SARS-CoV-2 on Highly Touched Surfaces in Brazil During the Second Wave of the COVID-19 Pandemic date: 2021-06-21 journal: nan DOI: 10.1101/2021.06.14.21258894 sha: 581e78abdc4258c58c25fd5ba47c21d7e1da8aa8 doc_id: 972103 cord_uid: 5zr5l1xl Although SARS-CoV-2 surface contamination has been investigated in temperate climates, few studies have been conducted in the tropics. Here, we investigated the presence of SARS-CoV-2 on high-touch surfaces in a large city in Brazil. A total of 400 surface samples were collected in February 2021 in the City of Recife, Northeastern Brazil. A total of 97 samples (24.2%) tested positive for SARS-CoV-2 by RT-qPCR using the CDC-USA protocol. All the collection sites, except one (18/19, 94.7%) had at least one environmental surface sample contaminated. SARS-CoV-2 positivity was higher in public transport terminals (47/97, 48.4%), followed by health care units (26/97, 26.8%), public parks (14/97, 14.4%), public markets (4/97, 4.1%), and beach areas (4/97, 4.1%). Toilets, ATMs, handrails, playground, and outdoor gym were identified as fomites with the highest rates of viral contamination. Regarding the type of material, SARS-CoV-2 RNA was found more commonly on metal (45/97, 46.3%), followed by plastic (18/97, 18.5%), wood (12/97, 12.3%), rock (10/97, 10.3%), concrete (8/97, 8.2%), and glass (2/97, 2.0%). Taken together, our data indicated extensive SARS-CoV-2 contamination in public surfaces and identified critical control points that need to be targeted to break SARS-CoV-2 transmission chains. (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint this version posted https://doi.org/10.1101 https://doi.org/10. /2021 Synopsis 48 We investigated the presence of SARS-CoV-2 on high-touch surfaces in a large 49 city in Brazil and identified critical points to establish effective control measures 50 aimed at breaking transmission. (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint this version posted June 21, 2021. ; https://doi.org/10.1101/2021.06.14.21258894 doi: medRxiv preprint (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint this version posted https://doi.org/10.1101 https://doi.org/10. /2021 Sampling areas 168 A total of 84 surface samples were collected from four public transport 170 terminals with a large daily passenger flow and concentration. We strategically 171 selected transport terminals that connect several cities in the metropolitan 172 region of Recife. Twenty-one swabs were collected for each transport terminal. 173 The collection points included the external area of the transport terminal and A total of 105 surface samples were collected from five public parks. We 187 strategically selected parks with high visitor flow, including children who access 188 the playground. Twenty-one swabs were collected for each public park. The 189 collection points included: (1) playground; (2) recreation area; (3) outdoor gym; 190 All rights reserved. No reuse allowed without permission. (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint this version posted https://doi.org/10.1101 https://doi.org/10. /2021 (4) toilet; (5) handrail; (6) bus stop; (7) public bike station; (8) traffic light button; 191 (9) coffee shop; (10) faucet. 192 A total of 85 surface samples were collected from four public markets. 194 Twenty-one swabs were collected for each public market with exception of one, 195 where we collected twenty-two swabs. The collection points included: (1) 196 principal entrance; (2) side entrance; (3) public market access; (4) toilet; (5) 197 kiosk; (6) store; (7) food hall; (8) (3) public bike station; (4) outdoor gym; (5) fresh green coconut; (6) handrails; 206 (7) faucet; (8) traffic light button; (9) bus stop; (10) resting area. 207 A total of 21 surface samples were collected from one food distribution 209 center located in Recife, Brazil. We selected this place as it is a place which 210 serves as a gateway for people from all over the Brazilian territory, and acts as 211 a source of food supply for the Northeast of Brazil. The collection points 212 included: (1) toilet; (2) restaurant; (3) handrail; (4) resting area. 213 All rights reserved. No reuse allowed without permission. (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. supplemented with 10% heat-inactivated fetal bovine serum, 100 U/ml penicillin 260 All rights reserved. No reuse allowed without permission. (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint this version posted June 21, 2021. ; https://doi.org/10.1101/2021.06.14.21258894 doi: medRxiv preprint 14 and 100 μg/ml streptomycin (Gibco, USA); and maintained maintained in a 261 humidified atmosphere, at 37 °C and 5% CO2. 262 Vero CCL-81 cells were cultured in 12-well plates at a density of 2 x 10 5 264 cells/well. After 24h, the culture media was removed and cells were incubated 265 with 300 µL of undiluted and filtered surface samples at 37°C, 5% CO2, for 1h. 266 Fresh media supplemented with 2% FBS (700 µL) was added to the cells and 267 they were maintained at 37°C, 5% CO 2 . Cells were monitored daily for the prevention measures, including social distancing, mask wearing, the availability 283 of hand sanitizers, body temperature measurements for screening and the 284 All rights reserved. No reuse allowed without permission. (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint this version posted June 21, 2021. ; https://doi.org/10.1101/2021.06.14.21258894 doi: medRxiv preprint 15 presence of informative charts for people education. The questionnaires were 285 made with qualitative, with "yes" or "no" input, or quantitative inquiries. 286 To georeference the locations where surface samples were obtained, we 288 (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint this version posted June 21, 2021. (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (Figure 3c ). Positive samples were predominantly found in 350 toilets, ATMs, handrails, playground, and outdoor gym; highlighting the 351 importance of these fomites in SARS-CoV-2 surface contamination. 352 All rights reserved. No reuse allowed without permission. (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint this version posted June 21, 2021. ; https://doi.org/10.1101/2021.06.14.21258894 doi: medRxiv preprint (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. Three out of four public markets sampled returned at least one positive 391 sample. Four (4.1%) surface samples were positive for SARS-CoV-2 RNA in 392 All rights reserved. No reuse allowed without permission. (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint this version posted June 21, 2021. ; https://doi.org/10.1101/2021.06.14.21258894 doi: medRxiv preprint identified in plastic surfaces, especially from traffic light buttons, near bus stops, 441 and in the toilets. Four (15.3%) samples were detected in rock surfaces found at 442 the entrance to hospitals. Two (7.6%) samples were identified in wood surfaces 443 at the entrance to hospitals. One (3.8%) sample was detected on the concrete 444 surface from a nearby bus stop ( Figure 5, Supplementary Table 3) . (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (1/19) of collection sites were limiting the number of people who accessed the 495 location point (Table 1) . We found no positive correlation between adherence of 496 COVID-19 mitigation measures and SARS-CoV-2 positivity (data not shown). 497 All rights reserved. No reuse allowed without permission. (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint this version posted June 21, 2021. ; https://doi.org/10.1101/2021.06.14.21258894 doi: medRxiv preprint (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint this version posted June 21, 2021. ; https://doi.org/10.1101/2021.06.14.21258894 doi: medRxiv preprint Since the emergence of SARS-CoV-2, first identified in China, the highly 518 pathogenic coronavirus has spread rapidly around the world causing an 519 unprecedented health security crisis and drastically affecting the global 520 economic stability. Thus, understanding the modes of transmission of SARS-521 CoV-2 among humans is a critical step to establish effective prevention policies 522 and prioritize resources to break the chain of SAR-CoV-2 transmission. The (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint this version posted https://doi.org/10.1101 https://doi.org/10. /2021 29 (5.25%) of surface samples were tested positive for SARS-CoV-2 RNA, 542 although the infectious potential of positive samples was not investigated. 543 Considering the proportion of positivity in the different places, the authors 544 pointed out that bus terminals exhibited the highest positivity rate, followed by 545 public markets, public squares, and health care units 49 Regarding the distribution of positive samples according to the type of 562 material, we found the SARS-CoV-2 RNA mainly on metal, followed by plastic, 563 wood, rock, concrete, and glass. Similarly, a recent urban study found the 564 SARS-CoV-2 RNA on different types of materials, the majority on metal, 565 concrete, rock, brickwork, wood, and glass 49 . Interestingly, our data 566 All rights reserved. No reuse allowed without permission. (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint this version posted https://doi.org/10.1101 https://doi.org/10. /2021 demonstrated that the positive samples for SARS-CoV-2 RNA were mainly 567 collected in toilets. These findings also corroborate data obtained by other 568 groups 26, 30, 53 , which toilets as an area of high positivity rate for SARS-CoV-2 569 RNA. Additionally, our findings revealed other specific locations with high rates 570 of positivity: ATMs, handrails, playgrounds, and outdoor gyms. 571 Previous studies performed under controlled laboratory conditions have 572 shown that SARS-CoV-2 remains infectious on different types of surfaces, such 573 as stainless steel, glass and paper, for up to 28 days at 20 °C 34 , depending on 574 type of environmental surface; and can remain viable in aerosols for up to 3 h 575 35 . Notably, the viral load decreases over time and depends on the length of 576 time since the virus has been deposited on the surface, which may be reflected 577 in the presence of infectious or non-infectious viral particles and, consequently, 578 infection risk in humans 21, 23, 24 . Another important factor that must be 579 considered is the minimal infectious dose of SARS-CoV-2 to start an effective 580 infection in humans, which has not yet been clarified. In order to elucidate the 581 transmission dynamics of SARS-CoV-2 by environmental surfaces in real-life 582 conditions, several studies have investigated the presence of SARS-CoV-2 in 583 air and environmental surfaces/areas, including health care settings [25] [26] [27] [28] [29] [30] [31] 33 and 584 urban settings 49, 52-55 . In general, these studies have found varying levels of 585 environmental contamination, ranging from extensive 25, 26 to low contamination 586 31, 49 , or even no contamination of SARS-CoV-2 RNA. However, many of these 587 studies did not determine the ability of SARS-CoV-2 to be cultured from such 588 environmental swabs, which would help to understand the implications of 589 SARS-CoV-2 RNA positive environmental samples in terms of infectious 590 potential for the human population 25, 27 . In this study, we evaluated the 591 All rights reserved. No reuse allowed without permission. (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint this version posted June 21, 2021. ; https://doi.org/10.1101/2021.06.14.21258894 doi: medRxiv preprint infectious potential of positive surface samples (Cq value <34) in Vero CCL-81 592 cells, but SARS-CoV-2 could not be cultured. This finding is supported by 593 recent studies, which have demonstrated the low potential infectious from the 594 environmental swabs using cell culture 25, 31, 56 considerably less efficient than direct contact transmission, whereby direct 615 All rights reserved. No reuse allowed without permission. (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint this version posted June 21, 2021. ; https://doi.org/10.1101/2021.06.14.21258894 doi: medRxiv preprint contacting animals are exposed to infected ferrets and share with them the 616 same food, water, bedding, and breathe the same air 42, 57 . 617 Regarding the adherence of COVID-19 mitigation measures by society, a 618 number of studies have been performed in order to evaluate the adoption of 619 measures to prevent the SARS-CoV-2 transmission 58-60 . To assess the 620 community's adherence to mitigation measures to combat the rapid spread of 621 SARS-CoV-2, a recent cross-sectional study conducted in Malaysia employed 622 4,850 Malaysian residents, between 27th March and 3rd April 2020 59 . The 623 findings revealed that most participants (83.1%) held positive attitudes toward 624 the successful control of COVID-19, the capacity of Malaysia to counter rapid 625 spread of the disease (95.9%) and the way the Malaysian government was 626 facing the COVID-19 crisis (89.9%). Furthermore, most participants were also 627 taking precautions such as practicing hand hygiene (87.8%) and avoiding large 628 gatherings (83.4%) 59 . Interestingly, the number of COVID-19 cases in Malaysia 629 remained stable, with a progressive increase observed only between 630 September and November 2020 (https://ourworldindata.org/covid-cases). In 631 contrast, a community-based cross-sectional study done in Northeast Ethiopia, 632 evaluated the adherence towards COVID-19 mitigation strategies by society 633 among 635 individuals from April 20-27, 2020 58 . The results showed that 634 approximately half of the study participants had poor adherence towards 635 COVID-19 mitigation measures. In the current analysis, although the number of 636 places evaluated was limited (19), it is important to highlight that these are 637 places with a high flow and concentration of people. Our data demonstrated low 638 adherence of COVID-19 mitigation measures by society regarding the social 639 distancing, effective use of masks, precaution measures adoption and 640 All rights reserved. No reuse allowed without permission. (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint this version posted June 21, 2021. ; https://doi.org/10.1101/2021.06.14.21258894 doi: medRxiv preprint (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint this version posted June 21, 2021. ; https://doi.org/10.1101/2021.06.14.21258894 doi: medRxiv preprint Coronaviruses: an overview of their replication and 685 pathogenesis Origin and evolution of pathogenic coronaviruses Epidemiology and 690 cause of severe acute respiratory syndrome (SARS) in Guangdong, People's Republic of China 693 Isolation of a novel coronavirus from a man with pneumonia in Saudi Arabia Genomic characterisation and 699 epidemiology of 2019 novel coronavirus: implications for virus origins and receptor binding A pneumonia outbreak associated with a new coronavirus 705 of probable bat origin A Novel Coronavirus from Patients with Pneumonia in China Clinical features of 713 patients infected with 2019 novel coronavirus in An interactive web-based dashboard to track COVID-19 716 in real time Genomics and epidemiology of the P.1 SARS-CoV-2 lineage in Manaus New SARS-CoV-2 Variants -Clinical, Public Health, 731 and Vaccine Implications COVID-19 in Amazonas, Brazil, was driven by the persistence of endemic 737 lineages and P.1 emergence Contamination of inert surfaces by SARS-CoV Characteristics of SARS-CoV-2 and COVID-19 Transmission 745 routes of respiratory viruses among humans Airflow dynamics of human 748 jets: sneezing and breathing -potential sources of infectious aerosols Respiratory virus shedding in exhaled breath and efficacy of face masks The airborne lifetime of small speech 755 droplets and their potential importance in SARS-CoV-2 transmission transmission, health impacts and mitigation: timely social distancing is the key Identifying airborne 761 transmission as the dominant route for the spread of COVID-19 Transmission routes for SARS-CoV-2 infection: review of 764 evidence Stability of SARS-CoV-2 in different environmental conditions Aerosol and Surface Stability of SARS-CoV-2 as Compared Investigating SARS-CoV-2 surface and air 774 contamination in an acute healthcare setting during the peak of the COVID Detection of air and surface contamination 780 by SARS-CoV-2 in hospital rooms of infected patients Aerosol and surface contamination of SARS-CoV-2 784 observed in quarantine and isolation care Environmental contamination of 787 SARS-CoV-2 on surfaces, air-conditioner and ventilation systems Air and surface contamination by SARS-CoV-2 virus in a tertiary 791 hospital in Wuhan, China Aerodynamic analysis of SARS-CoV-2 in 794 two Wuhan hospitals Severe acute 797 respiratory syndrome coronavirus 2 RNA contamination of inanimate surfaces and virus 798 viability in a health care emergency unit Environmental contamination of 801 SARS-CoV-2 in healthcare premises Investigation of SARS CoV-2 virus in 804 environmental surface The effect of temperature on 806 persistence of SARS-CoV-2 on common surfaces Aerosol and Surface Stability of SARS-CoV-2 as Compared with SARS Research use only 2019-Novel Coronavirus (2019-nCoV) Real-time RT-PCR 815 primers and probes WHO 39. Organization, W. H. Weekly epidemiological update -9 Circos: an information aesthetic for comparative genomics Ten 825 scientific reasons in support of airborne transmission of SARS-CoV-2 SARS-CoV-2 is transmitted via contact and via the air between ferrets Pathogenesis and transmission of SARS-CoV-2 in golden hamsters No reuse allowed without permission. (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint this version posted Transmission of Severe Acute Respiratory Syndrome 837 Coronavirus 2 via Close Contact and Respiratory Droplets Among Human Angiotensin-838 Converting Enzyme 2 Mice Infectious virus in exhaled breath of symptomatic 841 seasonal influenza cases from a college community Viable influenza A virus in airborne particles from human coughs 848 Evidence of Respiratory Syncytial Virus Spread by Aerosol. Time to Revisit Infection Control 849 Strategies? 851 Evidence of airborne transmission of the severe acute respiratory syndrome virus Detection of SARS-CoV-2 RNA on public surfaces in a densely populated urban 858 area of Brazil: A potential tool for monitoring the circulation of infected patients Humidity Accelerates Inactivation of SARS-CoV-2 on Surfaces COVID-19 transmission in Mainland China is associated with temperature and 867 humidity: A time-series analysis Longitudinal monitoring of SARS-CoV RNA on high-touch surfaces in a community setting. medRxiv 2020 Air and surface contamination in non-health care 873 settings among 641 environmental specimens of 39 COVID-19 cases Indirect Virus Transmission in 876 Cluster of COVID-19 Cases Air and 879 surface measurements of SARS-CoV-2 inside a bus during normal operation Low risk of 882 SARS-CoV-2 transmission by fomites in real-life conditions All rights reserved. No reuse allowed without permission. (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint this version posted Adherence towards COVID-19 mitigation 893 measures and its associated factors among Gondar City residents: A community-based cross-894 sectional study in Northwest Ethiopia Public knowledge, 896 attitudes and practices towards COVID-19: A cross-sectional study in Malaysia community's perception about the COVID-19 disease. Taken together, these 641 results highlight the importance of consistent messaging from government and 642 health authorities to improve levels the adoption of measures to prevent and 643 contain the spread of In summary, our data demonstrated the extensive viral RNA 645 contamination of surfaces in a range of public urban settings in the absence of 646 viral isolation, which suggests low potential risk from environmental 647 contamination for the human population. However, we identified poor 648 adherence to COVID-19 mitigation policies by wider society regarding the 649 adoption of control measures, and this may be reflected in the frequent 650 detection of the viral RNA. Studies such as these can contribute to assess the 651 prevalence of SARS-CoV-2 in specific settings. Finally, we suggest that further 652 studies are urgently performed to elucidate the relative contribution of various 653 modes of transmission for SARS-CoV-2 in both healthcare and urban-settings. 654 Acknowledgments 664 All rights reserved. No reuse allowed without permission.(which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity.The copyright holder for this preprint this version posted June 21, 2021. ; https://doi.org/10.1101/2021.06.14.21258894 doi: medRxiv preprint We are grateful to Msc. Larissa Krokovsky for her support and critical analysis 665 for the design of the work. We thank Josenildo do Sobral for helping to collect 666 surface samples. We would like to express our sincere gratitude and 667 appreciation to the Fiocruz Pernambuco team for the support. The authors declare no competing interests. 679 680 All rights reserved. No reuse allowed without permission.(which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity.The copyright holder for this preprint this version posted June 21, 2021. ; https://doi.org/10.1101/2021.06.14.21258894 doi: medRxiv preprint All rights reserved. No reuse allowed without permission.(which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity.The copyright holder for this preprint this version posted https://doi.org/10.1101 https://doi.org/10. /2021 All rights reserved. No reuse allowed without permission.(which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission.(which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity.The copyright holder for this preprint this version posted June 21, 2021. ; https://doi.org/10.1101/2021.06.14.21258894 doi: medRxiv preprint