Vibrio cholerae and Salmonella Typhi culture-based wastewater or non-sewered sanitation surveillance in a resource ... - Nature.com

Abstract

Background

In resource-limited regions, relying on individual clinical results to monitor community diseases is sometimes not possible. Establishing wastewater and non-sewered sanitation surveillance systems can offer opportunities to improve community health.

Objective

We provide our experience of establishing a wastewater and non-sewered sanitation surveillance laboratory in Malawi, a resource-limited region, for Vibrio cholerae and Salmonella serotype Typhi.

Methods

Three locations (inclusive of 8 discrete sample collection sites in total) in the Blantyre District were studied for nine weeks, from September 6 to November 1, 2022. Grab samples were collected weekly. We piloted locally available culture-based medical diagnostic methods for V. cholerae and S. Typhi in wastewater, followed by confirmation analysis of the isolates using reverse transcription polymerase chain reaction (RT-PCR).

Results

Bacterial counts ranged from up to 106 colony-forming units/mL for V. cholerae and up to 107 colony-forming units/mL for S. Typhi. RT-PCR of the isolates showed that the available culture-based medical diagnostic methods were successful in detecting V. cholerae but were less accurate for S. Typhi in wastewater.

Impact statement

This experience serves as a catalyst for the development and validation of alternative wastewater surveillance analytical methods that are not dependent solely on RT-PCR. In this field trial conducted in Africa, new data-driven approaches were developed to promote early-level wastewater research and expand analysis options in resource-limited settings. Although culture-based methods are labor-intensive and have some limitations, we suggest initially leveraging the overlap with the locally available medical testing capacity for V. cholerae, whereas S. Typhi with RT-PCR may still be required. Wastewater analysis may be acceptable for V. cholerae and S. Typhi, which have a high degree of clinical case underreporting, fecal shedding, short incubation periods, and clear outbreak trends, predominantly in low- and middle-income countries.

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Fig. 1: Sanitation system hospital manhole sampling site in Blantyre, Malawi.

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Data availability

Data generated in this study can be found in the article and its supplementary files.

References

  1. Muller M. Have five decades of development engineering research improved sanitation in Southern Africa? J Int Dev. 2020;32:96–111. https://doi.org/10.1002/jid.3452

    Article  Google Scholar 

  2. World Health Organization (WHO) and the United Nations Children's Fund (UNICEF). Progress on household drinking water, sanitation and hygiene 2000-2022. https://data.unicef.org/resources/jmp-report-2023/ Accessed 8 September 2023.

  3. Bustin SA, Benes V, Garson JA, Hellemans J, Huggett J, Kubista M, et al. The MIQE Guidelines: minimum information for publication of quantitative real-time PCR experiments. Clin Chem. 2009;55:611–22. https://doi.org/10.1373/clinchem.2008.112797

    Article  CAS  PubMed  Google Scholar 

  4. Capone D, Chigwechokha P, de los Reyes FL III, Holm RH, Risk BB, Tilley E, et al. Impact of sampling depth on pathogen detection in pit latrines. PLoS Negl Trop Dis. 2021;15:e0009176. https://doi.org/10.1371/journal.pntd.0009176

    Article  PubMed  PubMed Central  Google Scholar 

  5. Johnson R, Sharma JR, Ramharack P, Mangwana N, Kinnear C, Viraragavan A, et al. Tracking the circulating SARS-CoV-2 variant of concern in South Africa using wastewater-based epidemiology. Sci Rep. 2022;12:1–2. https://doi.org/10.1038/s41598-022-05110-4

    Article  CAS  Google Scholar 

  6. Mangwana N, Archer E, Muller CJ, Preiser W, Wolfaardt G, Kasprzyk-Hordern B, et al. Sewage surveillance of SARS-CoV-2 at student campus residences in the Western Cape, South Africa. Sci Total Environ. 2022;851:158028. https://doi.org/10.1016/j.scitotenv.2022.158028

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Servetas SL, Parratt KH, Brinkman NE, Shanks OC, Smith T, Mattson PJ, et al. Standards to support an enduring capability in wastewater surveillance for public health: Where are we? Case Stud Chem Environ Eng. 2022;6:100247. https://doi.org/10.1016/j.cscee.2022.100247

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Naughton CC, Holm RH, Lin NJ, James BP, Smith T. Online dashboards for SARS-CoV-2 wastewater data need standard best practices: an environmental health communication agenda. J Water Health. 2023;21:615–24. https://doi.org/10.2166/wh.2023.312

    Article  PubMed  Google Scholar 

  9. Bengtsson L, Gaudart J, Lu X, Moore S, Wetter E, Sallah K, et al. Using mobile phone data to predict the spatial spread of cholera. Sci Rep. 2015;5:1–5. https://doi.org/10.1038/srep08923

    Article  CAS  Google Scholar 

  10. Ali M, Nelson AR, Lopez AL, Sack DA. Updated global burden of cholera in endemic countries. PLoS Negl Trop Dis. 2015;9:e0003832. https://doi.org/10.1371/journal.pntd.0003832

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Kanungo S, Azman AS, Ramamurthy T, Deen J, Dutta S. Cholera. Lancet. 2022;399:1429–40. https://doi.org/10.1016/S0140-6736(22)00330-0

    Article  PubMed  Google Scholar 

  12. Stanaway JD, Reiner RC, Blacker BF, Goldberg EM, Khalil IA, Troeger CE, et al. The global burden of typhoid and paratyphoid fevers: a systematic analysis for the Global Burden of Disease Study 2017. Lancet Infect Dis. 2019;19:369–81. https://doi.org/10.1016/S1473-3099(18)30685-6

    Article  Google Scholar 

  13. Srinivasan M, Sindhu KN, Giri S, Kumar N, Mohan VR, Grassly NC, et al. Salmonella Typhi shedding and household transmission by children with blood culture-confirmed typhoid fever in Vellore, South India. J Infect Dis. 2021;224:S593–600. https://doi.org/10.1093/infdis/jiab409

    Article  PubMed  PubMed Central  Google Scholar 

  14. Feasey NA, Gaskell K, Wong V, Msefula C, Selemani G, Kumwenda S, et al. Rapid emergence of multidrug resistant, H58-lineage Salmonella typhi in Blantyre, Malawi. PLoS Negl Trop Dis. 2015;9:e0003748. https://doi.org/10.1371/journal.pntd.0003748

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Manor Y, Handsher R, Halmut T, Neuman M, Bobrov A, Rudich H, et al. Detection of poliovirus circulation by environmental surveillance in the absence of clinical cases in Israel and the Palestinian authority. J Clin Microbiol. 1999;37:1670–5. https://doi.org/10.1128/JCM.37.6.1670-1675.1999

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Asghar H, Diop OM, Weldegebriel G, Malik F, Shetty S, El Bassioni L, et al. Environmental surveillance for polioviruses in the Global Polio Eradication Initiative. J Infect Dis. 2014;210:S294–303. https://doi.org/10.1093/infdis/jiu384

    Article  PubMed  Google Scholar 

  17. Liu P, Ibaraki M, Kapoor R, Amin N, Das A, Miah R, et al. Development of Moore swab and ultrafiltration concentration and detection methods for Salmonella Typhi and Salmonella Paratyphi A in wastewater and application in Kolkata, India and Dhaka, Bangladesh. Front Microbiol. 2021;12:684094. https://doi.org/10.3389/fmicb.2021.684094

    Article  PubMed  PubMed Central  Google Scholar 

  18. Basu P, Choudhury S, Shridhar V, Huilgol P, Roychoudhury S, Nandi I, et al. Surveillance of SARS-CoV-2 RNA in open-water sewage canals contaminated with untreated wastewater in resource-constrained regions. Access Microbiol. 2022;4:000318. https://doi.org/10.1099/acmi.0.000318

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Jakariya M, Ahmed F, Islam MA, Al Marzan A, Hasan MN, Hossain M, et al. Wastewater-based epidemiological surveillance to monitor the prevalence of SARS-CoV-2 in developing countries with onsite sanitation facilities. Environ Pollut. 2022;311:119679. https://doi.org/10.1016/j.envpol.2022.119679

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Kolarević S, Micsinai A, Szántó-Egész R, Lukács A, Kračun-Kolarević M, Djordjevic A, et al. Wastewater-based epidemiology in countries with poor wastewater treatment—epidemiological indicator function of SARS-CoV-2 RNA in surface waters. Sci Total Environ. 2022;843:156964. https://doi.org/10.1016/j.scitotenv.2022.156964

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Chacón L, Morales E, Valiente C, Reyes L, Barrantes K. Wastewater-based epidemiology of enteric viruses and surveillance of acute gastrointestinal illness outbreaks in a resource-limited region. Am J Trop Med Hyg. 2021;105:1004–12. https://doi.org/10.4269/ajtmh.21-0050

    Article  PubMed  PubMed Central  Google Scholar 

  22. Zohra T, Ikram A, Salman M, Amir A, Saeed A, Ashraf Z, et al. Wastewater based environmental surveillance of toxigenic Vibrio cholerae in Pakistan. PLoS ONE. 2021;16:e0257414. https://doi.org/10.1371/journal.pone.0257414

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Zhou NA, Ong AQ, Fagnant-Sperati CS, Harrison JC, Kossik AL, Beck NK, et al. Evaluation of sampling and concentration methods for Salmonella enterica serovar Typhi detection from wastewater. Am J Trop Med Hyg. 2023;108:482.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. McCall C, Wu H, Miyani B, Xagoraraki I. Identification of multiple potential viral diseases in a large urban center using wastewater surveillance. Water Res. 2020;184:116160. https://doi.org/10.1016/j.watres.2020.116160

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Holm RH, Chigwechokha P, Kinnear C, Winters A, Street R. Promoting surveillance in sub-Saharan Africa: Moving to wastewater and environmental genomic surveillance requires more attention. ACS EST Water. 2023;3:1994–6. https://doi.org/10.1021/acsestwater.3c00314

    Article  CAS  Google Scholar 

  26. Parsons LM, Somoskövi Á, Gutierrez C, Lee E, Paramasivan CN, Abimiku AL, et al. Laboratory diagnosis of tuberculosis in resource-poor countries: challenges and opportunities. Clin Microbiol Rev. 2011;24:314–50. https://doi.org/10.1128/CMR.00059-10

    Article  PubMed  PubMed Central  Google Scholar 

  27. Ssengooba W, Gelderbloem SJ, Mboowa G, Wajja A, Namaganda C, Musoke P, et al. Feasibility of establishing a biosafety level 3 tuberculosis culture laboratory of acceptable quality standards in a resource-limited setting: an experience from Uganda. Health Res Policy Sy. 2015;13:1–10. https://doi.org/10.1186/1478-4505-13-4

    Article  Google Scholar 

  28. Crane PE, Silliman SE. Sampling strategies for estimation of parameters related to ground water quality. Groundwater. 2009;47:699–708.

    Article  CAS  Google Scholar 

  29. Knudsen BE, Bergmark L, Munk P, Lukjancenko O, Priemé A, Aarestrup FM, et al. Impact of sample type and DNA isolation procedure on genomic inference of microbiome composition. MSystems. 2016;1:e00095–16. https://doi.org/10.1128/msystems.00095-16

    Article  PubMed  PubMed Central  Google Scholar 

  30. Park JY, Jeon S, Kim JY, Park M, Kim S. Multiplex real-time polymerase chain reaction assays for simultaneous detection of Vibrio cholerae, Vibrio parahaemolyticus, and Vibrio vulnificus. Osong Public Health Res Perspect. 2013;4:133–9. https://doi.org/10.1016/j.phrp.2013.04.004

    Article  PubMed  PubMed Central  Google Scholar 

  31. Tennant SM, Toema D, Qamar F, Iqbal N, Boyd MA, Marshall JM, et al. Detection of typhoidal and paratyphoidal Salmonella in blood by real-time polymerase chain reaction. Clin Infect Dis. 2015;61:S241–50. https://doi.org/10.1093/cid/civ726

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Pilar AV, Petronella N, Dussault FM, Verster AJ, Bekal S, Levesque RC, et al. Similar yet different: phylogenomic analysis to delineate Salmonella and Citrobacter species boundaries. BMC Genom. 2020;21:1–3. https://doi.org/10.1186/s12864-020-06780-y

    Article  CAS  Google Scholar 

  33. Ministry of Health (MoH) [Malawi] and ICF International. Malawi service provision assessment (MSPA) 2013–14. Lilongwe: MoH and ICF International. 2014.

  34. Ngwira LG, Sharma B, Shrestha KB, Dahal S, Tuladhar R, Manthalu G, et al. Cost of wastewater-based environmental surveillance for SARS-CoV-2: evidence from pilot sites in Blantyre, Malawi and Kathmandu, Nepal. PLoS Glob Public Health. 2022;2:e0001377. https://doi.org/10.1371/journal.pgph.0001377

    Article  PubMed  PubMed Central  Google Scholar 

  35. Awofisayo-Okuyelu A, McCarthy N, Mgbakor I, Hall I. Incubation period of typhoidal salmonellosis: a systematic review and meta-analysis of outbreaks and experimental studies occurring over the last century. BMC Infect Dis. 2018;18:1–3. https://doi.org/10.1186/s12879-018-3391-3

    Article  Google Scholar 

  36. Sartwell PE. The incubation period of poliomyelitis. Am J Public Health. 1952;42:1403–8.

    Article  CAS  Google Scholar 

  37. Global Polio Eradication Initiative. Guidelines on environmental surveillance for detection of polioviruses. Global Polio Eradication Initiative. Geneva, Switzerland: World Health Organization; 2015. http://polioeradication.org/wp-content/uploads/2016/07/GPLN_GuidelinesES_April2015.pdf Accessed 7 September 2023.

  38. Nga TV, Karkey A, Dongol S, Thuy HN, Dunstan S, Holt K, et al. The sensitivity of real-time PCR amplification targeting invasive Salmonellaserovars in biological specimens. BMC Infect Dis. 2010;10:1–9. https://doi.org/10.1186/1471-2334-10-125

    Article  CAS  Google Scholar 

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Funding

This work was supported by grants from the Rockefeller Foundation, James Graham Brown Foundation, Owsley Brown II Family Foundation, and the Jon Rieger Seed Grant. The funders had no role in the study design, data collection and analysis, decision to publish, or manuscript preparation.

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