vestibИзвестия Национальной академии наук Беларуси. Серия биологических наукProceedings of the National Academy of Sciences of Belarus, Biological Series1029-89402524-230XThe Republican Unitary Enterprise Publishing House "Belaruskaya Navuka"10.29235/1029-8940-2022-67-4-386-397vestib-836Research ArticleСтатьиКачественный и количественный анализ вирусной контаминации сточных вод как элемент контроля за циркуляцией кишечных вирусов в Республике БеларусьQualitative and quantitative analysis of wastewater viral contamination as a control element for the circulation of enteric viruses in the Republic of Belarushttps://orcid.org/0000-0001-6431-5050ПоклонскаяН. В.PaklonskayaN. V.

Поклонская Наталья Владимировна – канд. биол. наук, вед. науч. сотрудник

ул. Филимонова, 23, 220114, г. Минск

Natallia V. Paklonskaya – Ph. D. (Biol.), Leading Researcher

23, Filimonov Str., 220114, Minsk

labsanvir@gmail.comhttps://orcid.org/0000-0001-7309-152XАмвросьеваТ. В.AmvrosievaT. V.

Амвросьева Тамара Васильевна – д-р мед. наук, профессор, заведующий лабораторией

ул. Филимонова, 23, 220114, г. Минск

Tamara V. Amvrosieva – D. Sc. (Med.), Professor, Head of the Laboratory

23, Filimonov Str., 220114, Minsk

amvrosieva@gmail.comhttps://orcid.org/0000-0002-6488-9422КолтуноваЮ. Б.KaltunovaY. B.

Колтунова Юлия Борисовна – мл. науч. сотрудник

ул. Филимонова, 23, 220114, г. Минск

Yuliya B. Kaltunova – Junior Researcher

23, Filimonov Str., 220114, Minsk

labsanvir@gmail.comhttps://orcid.org/0000-0003-4044-6827БельскаяИ. В.BelskayaI. V.

Бельская Инна Валерьевна – науч. сотрудник

ул. Филимонова, 23, 220114, г. Минск

Ina V. Belskaya – Researcher

23, Filimonov Str., 220114, Minsk

labsanvir@gmail.comhttps://orcid.org/0000-0002-4521-6599ШиловаЮ. А.ShilavaY. A.

Шилова Юлия Александровна – мл. науч. сотрудник

ул. Филимонова, 23, 220114, г. Минск

Yuliya A. Shilava – Junior Researcher

23, Filimonov Str., 220114, Minsk

labsanvir@gmail.comРеспубликанский научно-практический центр эпидемиологии и микробиологииRepublican Research and Practical Center for Epidemiology and Microbiology202201112022674386397Copyright © Поклонская Н.В., Амвросьева Т.В., Колтунова Ю.Б., Бельская И.В., Шилова Ю.А., 20222022Поклонская Н.В., Амвросьева Т.В., Колтунова Ю.Б., Бельская И.В., Шилова Ю.А.Paklonskaya N.V., Amvrosieva T.V., Kaltunova Y.B., Belskaya I.V., Shilava Y.A.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://vestibio.belnauka.by/jour/article/view/836

Эпидемиология сточных вод – новое направление исследований в системе надзора за циркуляцией кишечных вирусов. Целью настоящей работы был анализ результатов мониторинга сточных вод в Республике Беларусь в отношении кишечных вирусов в 2020–2021 гг. Детекция РНК ротавирусов А, норовирусов 1-й и 2-й геногрупп, энтеровирусов, ДНК аденовирусов F осуществлялась с помощью ОТ-ПЦР, определение количества нуклеиновых кислот – с применением количественной модификации метода, генотипирование возбудителей – путем секвенирования информативных для этого регионов генома вирусов. В 2020–2021 гг. из исследованных 1038 проб сточных вод 487 (46,9 %) содержали нуклеиновую кислоту (НК) одного или нескольких вирусов.  

В 2021 г. имело место снижение активности циркуляции большинства кишечных вирусов по сравнению с 2020 г., обусловленное, вероятно, более строгим соблюдением санитарно-гигиенических правил во время пандемии COVID-19. Преобладание в сточных водах ДНК аденовирусов F могло быть обусловлено их широким бессимптомным носительством и большей устойчивостью ДНК возбудителя во внешней среде. Количественный анализ содержания вирусных НК в сточной воде показал, что большинство проб с высокими концентрациями НК кишечных вирусов были отобраны из точек, куда осуществлялся сброс сточных вод непосредственно из учреждений здравоохранения. По результатам молекулярного типирования в исследованных сточных водах присутствовали наиболее распространенные гено- или серотипы возбудителей – норовирус GII.4[P16], аденовирус 41-го типа, энтеровирусы Коксаки В3, В5, ЕСНО 30 и ЕСНО 13.  

Полученные данные, основанные на количественном анализе содержания кишечных вирусов в сточных водах, представляют ценную информацию об их циркуляции в человеческой популяции, которая позволяет оценивать текущую эпидситуацию и прогнозировать ее развитие в ближайшей перспективе.

Wastewater epidemiology is a new field of research that has great potential in the system of enteric virus surveillance. The purpose of the presented study was to analyze the results of wastewater monitoring of enteric viruses in 2020– 2021 in the Republic of Belarus. Detection and quantification of nucleic acids of rotaviruses A, noroviruses 1 and 2, enteroviruses and adenoviruses F were carried out using a quantitative real time RT-PCR followed by nucleic acid sequence genotyping. In 2020–2021, 487 (46.9 %) of 1038 wastewater samples contained nucleic acid of one or more viruses. The results indicated that there was a decrease in the circulating activity of the most enteric viruses in 2021 compared to 2020, likely due to stricter hygiene practices during the COVID-19 pandemic. Predominance of adenoviruses F in wastewater could be associated with their wide asymptomatic spread, on the one hand, and with greater resistance of pathogen DNA in the external environment, on the other. The quantitative analysis showed that the highest viral nucleic acid concentrations were found in hospital wastewater samples. According to the results of molecular typing, wastewater contained norovirus GII.4 [P16], adenovirus type 41, Coxsackievirus B3, B5 and Echovirus 13, the most common genotypes of enteric viruses in human population.  

Quantitative analysis and genotyping of nucleic acids of viruses from wastewater can be a source of valuable information for tracking their circulation in the human population to assess the current epidemiological situation and to predict its development in the short term.

ротавирус Аноровирусаденовирус Fэнтеровируссточная водамониторингзаболеваемостьэпидемиологияrotavirus Anorovirusadenovirus FenteroviruswastewatersurveillancemorbidityepidemiologyReferencesDaughton, C. Illicit drugs in municipal sewage: Proposed new non-intrusive tool to heighten public awareness of societal use of illicit/abused drugs and their potential for ecological consequences / C. Daughton // Pharmaceuticals and Personal Care Products in the Environment: Scientific and Regulatory Issues Edition: Symposium Series 791. – Washington : Am. Chem. Soc., 2001. – Ch. 20. – P. 348–364.Daughton, C. Illicit drugs in municipal sewage: Proposed new non-intrusive tool to heighten public awareness of societal use of illicit/abused drugs and their potential for ecological consequences / C. Daughton // Pharmaceuticals and Personal Care Products in the Environment: Scientific and Regulatory Issues Edition: Symposium Series 791. – Washington : Am. Chem. Soc., 2001. – Ch. 20. – P. 348–364.Wastewater-based epidemiology as a novel biomonitoring tool to evaluate human exposure to pollutants / E. GraciaLor [et al.] // Environ. Sci. Technol. – 2018. – Vol. 52, N 18. – P. 10224–10226. https://doi.org/10.1021/acs.est.8b01403Wastewater-based epidemiology as a novel biomonitoring tool to evaluate human exposure to pollutants / E. GraciaLor [et al.] // Environ. Sci. Technol. – 2018. – Vol. 52, N 18. – P. 10224–10226. https://doi.org/10.1021/acs.est.8b01403Sims, N. Future perspectives of wastewater-based epidemiology: Monitoring infectious disease spread and resistance to the community level // N. Sims, B. Kasprzyk-Hordern // Environ. Int. – 2020. – Vol. 139. – Art. 105689. https://doi.org/ 10.1016/j.envint.2020.105689Sims, N. Future perspectives of wastewater-based epidemiology: Monitoring infectious disease spread and resistance to the community level // N. Sims, B. Kasprzyk-Hordern // Environ. Int. – 2020. – Vol. 139. – Art. 105689. https://doi.org/ 10.1016/j.envint.2020.105689Xagoraraki, I. Wastewater-based epidemiology for early detection of viral outbreaks / I. Xagoraraki, E. O’Brien // Women in Water Quality: Investigations by Prominent Female Engineers / ed. D. O’Bannon. – Cham, 2019. – P. 75–97.Xagoraraki, I. Wastewater-based epidemiology for early detection of viral outbreaks / I. Xagoraraki, E. O’Brien // Women in Water Quality: Investigations by Prominent Female Engineers / ed. D. O’Bannon. – Cham, 2019. – P. 75–97.Occurrence of various viruses and recent evidence of SARS-CoV-2 in wastewater systems / W. Ali [et al.] // J. Hazard Mater. – 2021. – Vol. 414. – Art. 125439. https://doi.org/10.1016/j.jhazmat.2021.125439Occurrence of various viruses and recent evidence of SARS-CoV-2 in wastewater systems / W. Ali [et al.] // J. Hazard Mater. – 2021. – Vol. 414. – Art. 125439. https://doi.org/10.1016/j.jhazmat.2021.125439Wastewater-based epidemiology as an early warning system for the spreading of SARS-CoV-2 and its mutations in the population / T. Mackuľak [et al.] // Int. J. Environ. Res. Publ. Health. – 2021. – Vol. 18, N 11. – Art. 5629. https://doi. org/10.3390/ijerph18115629Wastewater-based epidemiology as an early warning system for the spreading of SARS-CoV-2 and its mutations in the population / T. Mackuľak [et al.] // Int. J. Environ. Res. Publ. Health. – 2021. – Vol. 18, N 11. – Art. 5629. https://doi. org/10.3390/ijerph18115629Norovirus genotypes present in oysters and in effluent from a wastewater treatment plant during the seasonal peak of infections in Ireland in 2010 / P. Rajko-Nenow [et al.] // Appl. Environ. Microbiol. – 2013. – Vol. 79, N 8. – P. 2578–2587. https://doi.org/10.1128/aem.03557-12Norovirus genotypes present in oysters and in effluent from a wastewater treatment plant during the seasonal peak of infections in Ireland in 2010 / P. Rajko-Nenow [et al.] // Appl. Environ. Microbiol. – 2013. – Vol. 79, N 8. – P. 2578–2587. https://doi.org/10.1128/aem.03557-12Environmental surveillance of norovirus genogroups i and ii for sensitive detection of epidemic variants / S. Kazama [et al.] // Appl. Environ. Microbiol. – 2017. – Vol. 83, N 9. – P. e03406-16. https://doi.org/10.1128/aem.03406-16Environmental surveillance of norovirus genogroups i and ii for sensitive detection of epidemic variants / S. Kazama [et al.] // Appl. Environ. Microbiol. – 2017. – Vol. 83, N 9. – P. e03406-16. https://doi.org/10.1128/aem.03406-16Monitoring human enteric viruses in wastewater and relevance to infections encountered in the clinical setting: a one year experiment in central France, 2014 to 2015 / M. Bisseux [et al.] // Eurosurveillance. – 2018. – Vol. 23, N 7. – pii 17-00237. https://doi.org/10.2807/1560-7917.es.2018.23.7.17-00237Monitoring human enteric viruses in wastewater and relevance to infections encountered in the clinical setting: a one year experiment in central France, 2014 to 2015 / M. Bisseux [et al.] // Eurosurveillance. – 2018. – Vol. 23, N 7. – pii 17-00237. https://doi.org/10.2807/1560-7917.es.2018.23.7.17-00237Sanger, F. DNA sequencing with chain-terminating inhibitors / F. Sanger, S. Nicklen, A. R. Coulson // Proc. Natl. Acad. Sci. USA. – 1977. – Vol. 74, N 12. – P. 5463–5467. https://doi.org/10.1073/pnas.74.12.5463Sanger, F. DNA sequencing with chain-terminating inhibitors / F. Sanger, S. Nicklen, A. R. Coulson // Proc. Natl. Acad. Sci. USA. – 1977. – Vol. 74, N 12. – P. 5463–5467. https://doi.org/10.1073/pnas.74.12.5463Comparison of primers for the detection of genogroup II noroviruses in India / S. George [et al.] // Indian J. Med. Microbiol. – 2012. – Vol. 30, N 1. – P. 24–29. https://doi.org/10.4103/0255-0857.93016Comparison of primers for the detection of genogroup II noroviruses in India / S. George [et al.] // Indian J. Med. Microbiol. – 2012. – Vol. 30, N 1. – P. 24–29. https://doi.org/10.4103/0255-0857.93016Genogroup-specific PCR primers for detection of Norwalk-like viruses / S. Kojima [et al.] // J. Virol. Methods. – 2002. – Vol. 100, N 1–2. – P. 107–114. https://doi.org/10.1016/s0166-0934(01)00404-9Genogroup-specific PCR primers for detection of Norwalk-like viruses / S. Kojima [et al.] // J. Virol. Methods. – 2002. – Vol. 100, N 1–2. – P. 107–114. https://doi.org/10.1016/s0166-0934(01)00404-9Development of enhanced primer sets for detection of norovirus / B. H. Kong [et al.] // Biomed. Res. Int. – 2015. – Vol. 2015. – P. 1–9. https://doi.org/10.1155/2015/103052Development of enhanced primer sets for detection of norovirus / B. H. Kong [et al.] // Biomed. Res. Int. – 2015. – Vol. 2015. – P. 1–9. https://doi.org/10.1155/2015/103052Lu, X. Molecular typing of human adenoviruses by PCR and sequencing of a partial region of the hexon gene / X. Lu, D. D. Erdman // Arch. Virol. – 2006. – Vol. 151, N 8. – P. 1587–1602. https://doi.org/10.1007/s00705-005-0722-7Lu, X. Molecular typing of human adenoviruses by PCR and sequencing of a partial region of the hexon gene / X. Lu, D. D. Erdman // Arch. Virol. – 2006. – Vol. 151, N 8. – P. 1587–1602. https://doi.org/10.1007/s00705-005-0722-7Prospective identification of HEV-B enteroviruses during the 2005 outbreak / A. Mirand [et al.] // J. Med. Virol. – 2006. – Vol. 78, N 12. – P. 1624–1634. https://doi.org/10.1002/jmv.20747Prospective identification of HEV-B enteroviruses during the 2005 outbreak / A. Mirand [et al.] // J. Med. Virol. – 2006. – Vol. 78, N 12. – P. 1624–1634. https://doi.org/10.1002/jmv.20747Nix, W. A. Sensitive, seminested PCR amplification of VP1 sequences for direct identification of all enterovirus serotypes from original clinical specimens / W. A. Nix, M. S. Oberste, M. A. Pallansch // J. Clin. Microbiol. – 2006. – Vol. 44, N 8. – P. 2698–2704. https://doi.org/10.1128/jcm.00542-06Nix, W. A. Sensitive, seminested PCR amplification of VP1 sequences for direct identification of all enterovirus serotypes from original clinical specimens / W. A. Nix, M. S. Oberste, M. A. Pallansch // J. Clin. Microbiol. – 2006. – Vol. 44, N 8. – P. 2698–2704. https://doi.org/10.1128/jcm.00542-06Basic local alignment search tool / S. F. Altschul [et al.] // J. Mol. Biol. – 1990. – Vol. 215, N 3. – P. 403–410. https:// doi.org/10.1016/S0022-2836(05)80360-2Basic local alignment search tool / S. F. Altschul [et al.] // J. Mol. Biol. – 1990. – Vol. 215, N 3. – P. 403–410. https:// doi.org/10.1016/S0022-2836(05)80360-2Kumar, S. MEGA7: Molecular Evolutionary Genetics Analysis Version 7.0 for Bigger Datasets / S. Kumar, G. Stecher, K. Tamura // Mol. Biol. Evol. – 2016. – Vol. 33, N 7. – P. 1870–1874. https://doi.org/10.1093/molbev/msw054Kumar, S. MEGA7: Molecular Evolutionary Genetics Analysis Version 7.0 for Bigger Datasets / S. Kumar, G. Stecher, K. Tamura // Mol. Biol. Evol. – 2016. – Vol. 33, N 7. – P. 1870–1874. https://doi.org/10.1093/molbev/msw054An automated genotyping tool for enteroviruses and noroviruses / A. Kroneman [et al.] // J. Clin. Virol. – 2011. – Vol. 51, N 2. – P. 121–125. https://doi.org/10.1016/j.jcv.2011.03.006An automated genotyping tool for enteroviruses and noroviruses / A. Kroneman [et al.] // J. Clin. Virol. – 2011. – Vol. 51, N 2. – P. 121–125. https://doi.org/10.1016/j.jcv.2011.03.006Гланц, С. Медико-биологическая статистика / С. Гланц. – М. : Практика, 1998. – 459 с.Glants S. Medico-biological statistics. Moscow, Praktika Publ., 1998. 459 p. (in Russian).Possible indicator role of adenoviruses for assessing viral contamination of water / T. V. Amvrosieva [et al.] // Proc. of the 11th Eastern European young water professional conference, 01–05.10.2019, Praha / Univ. of Chem. and Technol. – Praha, 2019. – P. 76–81.Amvrosieva T. V., Paklonskaya N. V., Belskaya I. V., Laziuk S. K., Kazinets O. N., Shilova Yu. A. Possible indicator role of adenoviruses for assessing viral contamination of water. Proceedings of the 11th Eastern European young water professional conference, 01–05.10.2019, Praha. Praha, 2019, pp. 76–81.МУК 4.2.2029-05. Санитарно-вирусологический контроль водных объектов : метод. указания. – Введ. : 2005-11-18. – М. : Роспотребнадзор, 2006. – 40 с.MUK 4.2.2029-05. Sanitary and virological control of water bodies: method. instructions. Introduced: 2005-11-18. Moscow, Rospotrebnadzor Publ., 2006. 40 p. (in Russian).One-year monthly survey of rotavirus, astrovirus and norovirus in three sewage treatment plants (STPs) in Beijing, China and associated health risk assessment / X. Q. He [et al.] // Water Sci. Technol. – 2011. – Vol. 64, N 6. – P. 1202–1210. https://doi.org/10.2166/wst.2011.080One-year monthly survey of rotavirus, astrovirus and norovirus in three sewage treatment plants (STPs) in Beijing, China and associated health risk assessment / X. Q. He [et al.] // Water Sci. Technol. – 2011. – Vol. 64, N 6. – P. 1202–1210. https://doi.org/10.2166/wst.2011.080Quantification and molecular characterization of enteric viruses detected in effluents from two hospital wastewater treatment plants / T. Prado [et al.] // Water Res. – 2011. – Vol. 45, N 3. – P. 1287–1297. https://doi.org/10.1016/j.watres.2010.10.012Quantification and molecular characterization of enteric viruses detected in effluents from two hospital wastewater treatment plants / T. Prado [et al.] // Water Res. – 2011. – Vol. 45, N 3. – P. 1287–1297. https://doi.org/10.1016/j.watres.2010.10.012Detection and quantitation of infectious human adenoviruses and JC polyomaviruses in water by immunofluo rescence assay / B. Calgua [et al.] // J. Virol. Methods. – 2011. – Vol. 171, N 1. – P. 1–7. https://doi.org/10.1016/j.jviromet. 2010.09.013Detection and quantitation of infectious human adenoviruses and JC polyomaviruses in water by immunofluo rescence assay / B. Calgua [et al.] // J. Virol. Methods. – 2011. – Vol. 171, N 1. – P. 1–7. https://doi.org/10.1016/j.jviromet. 2010.09.013UV inactivation of human infectious viruses at two full-scale wastewater treatment plants in Canada / Y. Qiu [et al.] // Water Res. – 2018. – Vol. 147. – P. 73–81. https://doi.org/10.1016/j.watres.2018.09.057UV inactivation of human infectious viruses at two full-scale wastewater treatment plants in Canada / Y. Qiu [et al.] // Water Res. – 2018. – Vol. 147. – P. 73–81. https://doi.org/10.1016/j.watres.2018.09.057High occurrence of hepatitis E virus in samples from wastewater treatment plants in Switzerland and comparison with other enteric viruses / F. G. Masclaux [et al.] // Water Res. – 2013. – Vol. 47, N 14. – P. 5101–5109. https://doi.org/10.1016/j. watres.2013.05.050High occurrence of hepatitis E virus in samples from wastewater treatment plants in Switzerland and comparison with other enteric viruses / F. G. Masclaux [et al.] // Water Res. – 2013. – Vol. 47, N 14. – P. 5101–5109. https://doi.org/10.1016/j. watres.2013.05.050Qualitative and quantitative molecular detection of enteroviruses in water from bathing areas and from a sewage treatment plant / E. Schvoerer [et al.] // Res. Microbiol. – 2001. – Vol. 52, N 2. – P. 179–186. https://doi.org/10.1016/s0923- 2508(01)01190-1Qualitative and quantitative molecular detection of enteroviruses in water from bathing areas and from a sewage treatment plant / E. Schvoerer [et al.] // Res. Microbiol. – 2001. – Vol. 52, N 2. – P. 179–186. https://doi.org/10.1016/s0923- 2508(01)01190-1

The authors declare that there are no conflicts of interest present.