Diverse lineages of pathogenic Leptospira species are widespread in the environment in Puerto Rico, USA.

<h4>Background</h4>Leptospirosis, caused by Leptospira bacteria, is a common zoonosis worldwide, especially in the tropics. Reservoir species and risk factors have been identified but surveys for environmental sources are rare. Furthermore, understanding of environmental Leptospira conta...

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Main Authors: Nathan E Stone, Carina M Hall, Marielisa Ortiz, Shelby M Hutton, Ella Santana-Propper, Kimberly R Celona, Charles H D Williamson, Nicole Bratsch, Luis G V Fernandes, Joseph D Busch, Talima Pearson, Sarai Rivera-Garcia, Fred Soltero, Renee Galloway, Jason W Sahl, Jarlath E Nally, David M Wagner
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2022-05-01
Series:PLoS Neglected Tropical Diseases
Online Access:https://doi.org/10.1371/journal.pntd.0009959
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author Nathan E Stone
Carina M Hall
Marielisa Ortiz
Shelby M Hutton
Ella Santana-Propper
Kimberly R Celona
Charles H D Williamson
Nicole Bratsch
Luis G V Fernandes
Joseph D Busch
Talima Pearson
Sarai Rivera-Garcia
Fred Soltero
Renee Galloway
Jason W Sahl
Jarlath E Nally
David M Wagner
author_facet Nathan E Stone
Carina M Hall
Marielisa Ortiz
Shelby M Hutton
Ella Santana-Propper
Kimberly R Celona
Charles H D Williamson
Nicole Bratsch
Luis G V Fernandes
Joseph D Busch
Talima Pearson
Sarai Rivera-Garcia
Fred Soltero
Renee Galloway
Jason W Sahl
Jarlath E Nally
David M Wagner
author_sort Nathan E Stone
collection DOAJ
description <h4>Background</h4>Leptospirosis, caused by Leptospira bacteria, is a common zoonosis worldwide, especially in the tropics. Reservoir species and risk factors have been identified but surveys for environmental sources are rare. Furthermore, understanding of environmental Leptospira containing virulence associated genes and possibly capable of causing disease is incomplete, which may convolute leptospirosis diagnosis, prevention, and epidemiology.<h4>Methodology/principal findings</h4>We collected environmental samples from 22 sites in Puerto Rico during three sampling periods over 14-months (Dec 2018-Feb 2020); 10 water and 10 soil samples were collected at each site. Samples were screened for DNA from potentially pathogenic Leptospira using the lipL32 PCR assay and positive samples were sequenced to assess genetic diversity. One urban site in San Juan was sampled three times over 14 months to assess persistence in soil; live leptospires were obtained during the last sampling period. Isolates were whole genome sequenced and LipL32 expression was assessed in vitro. We detected pathogenic Leptospira DNA at 15/22 sites; both soil and water were positive at 5/15 sites. We recovered lipL32 sequences from 83/86 positive samples (15/15 positive sites) and secY sequences from 32/86 (10/15 sites); multiple genotypes were identified at 12 sites. These sequences revealed significant diversity across samples, including four novel lipL32 phylogenetic clades within the pathogenic P1 group. Most samples from the serially sampled site were lipL32 positive at each time point. We sequenced the genomes of six saprophytic and two pathogenic Leptospira isolates; the latter represent a novel pathogenic Leptospira species likely belonging to a new serogroup.<h4>Conclusions/significance</h4>Diverse and novel pathogenic Leptospira are widespread in the environment in Puerto Rico. The disease potential of these lineages is unknown but several were consistently detected for >1 year in soil, which could contaminate water. This work increases understanding of environmental Leptospira diversity and should improve leptospirosis surveillance and diagnostics.
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spelling doaj.art-be6400c8f2d94ade8b995883a0691efb2022-12-22T01:53:00ZengPublic Library of Science (PLoS)PLoS Neglected Tropical Diseases1935-27271935-27352022-05-01165e000995910.1371/journal.pntd.0009959Diverse lineages of pathogenic Leptospira species are widespread in the environment in Puerto Rico, USA.Nathan E StoneCarina M HallMarielisa OrtizShelby M HuttonElla Santana-PropperKimberly R CelonaCharles H D WilliamsonNicole BratschLuis G V FernandesJoseph D BuschTalima PearsonSarai Rivera-GarciaFred SolteroRenee GallowayJason W SahlJarlath E NallyDavid M Wagner<h4>Background</h4>Leptospirosis, caused by Leptospira bacteria, is a common zoonosis worldwide, especially in the tropics. Reservoir species and risk factors have been identified but surveys for environmental sources are rare. Furthermore, understanding of environmental Leptospira containing virulence associated genes and possibly capable of causing disease is incomplete, which may convolute leptospirosis diagnosis, prevention, and epidemiology.<h4>Methodology/principal findings</h4>We collected environmental samples from 22 sites in Puerto Rico during three sampling periods over 14-months (Dec 2018-Feb 2020); 10 water and 10 soil samples were collected at each site. Samples were screened for DNA from potentially pathogenic Leptospira using the lipL32 PCR assay and positive samples were sequenced to assess genetic diversity. One urban site in San Juan was sampled three times over 14 months to assess persistence in soil; live leptospires were obtained during the last sampling period. Isolates were whole genome sequenced and LipL32 expression was assessed in vitro. We detected pathogenic Leptospira DNA at 15/22 sites; both soil and water were positive at 5/15 sites. We recovered lipL32 sequences from 83/86 positive samples (15/15 positive sites) and secY sequences from 32/86 (10/15 sites); multiple genotypes were identified at 12 sites. These sequences revealed significant diversity across samples, including four novel lipL32 phylogenetic clades within the pathogenic P1 group. Most samples from the serially sampled site were lipL32 positive at each time point. We sequenced the genomes of six saprophytic and two pathogenic Leptospira isolates; the latter represent a novel pathogenic Leptospira species likely belonging to a new serogroup.<h4>Conclusions/significance</h4>Diverse and novel pathogenic Leptospira are widespread in the environment in Puerto Rico. The disease potential of these lineages is unknown but several were consistently detected for >1 year in soil, which could contaminate water. This work increases understanding of environmental Leptospira diversity and should improve leptospirosis surveillance and diagnostics.https://doi.org/10.1371/journal.pntd.0009959
spellingShingle Nathan E Stone
Carina M Hall
Marielisa Ortiz
Shelby M Hutton
Ella Santana-Propper
Kimberly R Celona
Charles H D Williamson
Nicole Bratsch
Luis G V Fernandes
Joseph D Busch
Talima Pearson
Sarai Rivera-Garcia
Fred Soltero
Renee Galloway
Jason W Sahl
Jarlath E Nally
David M Wagner
Diverse lineages of pathogenic Leptospira species are widespread in the environment in Puerto Rico, USA.
PLoS Neglected Tropical Diseases
title Diverse lineages of pathogenic Leptospira species are widespread in the environment in Puerto Rico, USA.
title_full Diverse lineages of pathogenic Leptospira species are widespread in the environment in Puerto Rico, USA.
title_fullStr Diverse lineages of pathogenic Leptospira species are widespread in the environment in Puerto Rico, USA.
title_full_unstemmed Diverse lineages of pathogenic Leptospira species are widespread in the environment in Puerto Rico, USA.
title_short Diverse lineages of pathogenic Leptospira species are widespread in the environment in Puerto Rico, USA.
title_sort diverse lineages of pathogenic leptospira species are widespread in the environment in puerto rico usa
url https://doi.org/10.1371/journal.pntd.0009959
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