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Human leptospirosis in Tanzania: sequencing and phylogenetic analysis confirm that pathogenic Leptospira species circulate among agro-pastoralists living in Katavi-Rukwa ecosystem

BACKGROUND: Leptospirosis is a neglected zoonotic disease of worldwide public health importance. The disease affects humans, domestic animals and wildlife. However, leptospirosis is challenging in its diagnosis in humans. Culture technique, which is time consuming, is not recommended for clinical di...

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Autores principales: Muller, Shabani K., Assenga, Justine A., Matemba, Lucas E., Misinzo, Gerald, Kazwala, Rudovick R.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4902944/
https://www.ncbi.nlm.nih.gov/pubmed/27287703
http://dx.doi.org/10.1186/s12879-016-1588-x
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author Muller, Shabani K.
Assenga, Justine A.
Matemba, Lucas E.
Misinzo, Gerald
Kazwala, Rudovick R.
author_facet Muller, Shabani K.
Assenga, Justine A.
Matemba, Lucas E.
Misinzo, Gerald
Kazwala, Rudovick R.
author_sort Muller, Shabani K.
collection PubMed
description BACKGROUND: Leptospirosis is a neglected zoonotic disease of worldwide public health importance. The disease affects humans, domestic animals and wildlife. However, leptospirosis is challenging in its diagnosis in humans. Culture technique, which is time consuming, is not recommended for clinical diagnosis. For these reasons, serological and molecular techniques remain the test of choice. The major objective of this study was to explore the genetic characteristic of Leptospira species which are prevalent among agro-pastoralists living in Katavi–Rukwa Ecosystem, Tanzania. METHODS: A cross-sectional epidemiological study was carried out in the Katavi-Region South-west, Tanzania between August, 2013 and November, 2014. A total of 267 participants were randomly recruited for the study. Microscopic agglutination test (MAT) was used to detect antibody against six Leptospira antigens including local serogroups Icterohaemorrhagiae, Ballum, Grippotyphosa, Sejroe and reference serogroups Hebdomadis, and Australis. Samples with MAT titers ≥ 1:160 were scored as positive, samples with MAT titers ranging from 1:20 to 1:80 were scored as exposed to Leptospira, and absence of agglutination titers was scored as negative. All MAT positive samples, including the low titre samples were subjected to PCR using the respective 16S rRNA primers for the pathogenic and non-pathogenic species. RESULTS: Out of 267 samples tested, 80 (29.9 %) were positive with MAT. The major circulating leptospiral serogroups were Sejroe (15.7 %,), Icterohaemorrhagiae (8.9 %), Grippotyphosa (4.8 %), Hebdomadis (3.37 %), Australis (1.49 %) and Ballum (1.19 %). By using PCR, 33 (15.7 %) out of 210 samples were pathogenic Leptospira while no saprophytic Leptospira spp. was detected. Partial 16S rRNA gene sequences of Leptospira species which were obtained from this study were submitted to GenBank and acquired accession numbers KP313246 and KP313247. Phylogenetic analysis of the nucleotide sequences revealed that species obtained from Katavi-Rukwa ecosystem clustered in the same group with several published pathogenic Leptospira specifically Leptospira interrogans and Leptospira kirschneri. To the best of the authors’ knowledge(,) this is the first study from Tanzania to confirm pathogenic Leptospira in human subjects using genomic typing technique. CONCLUSION: These findings provide ultimate evidence of pathogenic Leptospira species circulating among agro-pastoralists living in Katavi-Rukwa Ecosystem suggesting that active disease surveillance should be undertaken in order to achieve greater protection of the agro-pastoral communities in Tanzania. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12879-016-1588-x) contains supplementary material, which is available to authorized users.
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spelling pubmed-49029442016-06-13 Human leptospirosis in Tanzania: sequencing and phylogenetic analysis confirm that pathogenic Leptospira species circulate among agro-pastoralists living in Katavi-Rukwa ecosystem Muller, Shabani K. Assenga, Justine A. Matemba, Lucas E. Misinzo, Gerald Kazwala, Rudovick R. BMC Infect Dis Research Article BACKGROUND: Leptospirosis is a neglected zoonotic disease of worldwide public health importance. The disease affects humans, domestic animals and wildlife. However, leptospirosis is challenging in its diagnosis in humans. Culture technique, which is time consuming, is not recommended for clinical diagnosis. For these reasons, serological and molecular techniques remain the test of choice. The major objective of this study was to explore the genetic characteristic of Leptospira species which are prevalent among agro-pastoralists living in Katavi–Rukwa Ecosystem, Tanzania. METHODS: A cross-sectional epidemiological study was carried out in the Katavi-Region South-west, Tanzania between August, 2013 and November, 2014. A total of 267 participants were randomly recruited for the study. Microscopic agglutination test (MAT) was used to detect antibody against six Leptospira antigens including local serogroups Icterohaemorrhagiae, Ballum, Grippotyphosa, Sejroe and reference serogroups Hebdomadis, and Australis. Samples with MAT titers ≥ 1:160 were scored as positive, samples with MAT titers ranging from 1:20 to 1:80 were scored as exposed to Leptospira, and absence of agglutination titers was scored as negative. All MAT positive samples, including the low titre samples were subjected to PCR using the respective 16S rRNA primers for the pathogenic and non-pathogenic species. RESULTS: Out of 267 samples tested, 80 (29.9 %) were positive with MAT. The major circulating leptospiral serogroups were Sejroe (15.7 %,), Icterohaemorrhagiae (8.9 %), Grippotyphosa (4.8 %), Hebdomadis (3.37 %), Australis (1.49 %) and Ballum (1.19 %). By using PCR, 33 (15.7 %) out of 210 samples were pathogenic Leptospira while no saprophytic Leptospira spp. was detected. Partial 16S rRNA gene sequences of Leptospira species which were obtained from this study were submitted to GenBank and acquired accession numbers KP313246 and KP313247. Phylogenetic analysis of the nucleotide sequences revealed that species obtained from Katavi-Rukwa ecosystem clustered in the same group with several published pathogenic Leptospira specifically Leptospira interrogans and Leptospira kirschneri. To the best of the authors’ knowledge(,) this is the first study from Tanzania to confirm pathogenic Leptospira in human subjects using genomic typing technique. CONCLUSION: These findings provide ultimate evidence of pathogenic Leptospira species circulating among agro-pastoralists living in Katavi-Rukwa Ecosystem suggesting that active disease surveillance should be undertaken in order to achieve greater protection of the agro-pastoral communities in Tanzania. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12879-016-1588-x) contains supplementary material, which is available to authorized users. BioMed Central 2016-06-10 /pmc/articles/PMC4902944/ /pubmed/27287703 http://dx.doi.org/10.1186/s12879-016-1588-x Text en © Muller et al. 2016 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research Article
Muller, Shabani K.
Assenga, Justine A.
Matemba, Lucas E.
Misinzo, Gerald
Kazwala, Rudovick R.
Human leptospirosis in Tanzania: sequencing and phylogenetic analysis confirm that pathogenic Leptospira species circulate among agro-pastoralists living in Katavi-Rukwa ecosystem
title Human leptospirosis in Tanzania: sequencing and phylogenetic analysis confirm that pathogenic Leptospira species circulate among agro-pastoralists living in Katavi-Rukwa ecosystem
title_full Human leptospirosis in Tanzania: sequencing and phylogenetic analysis confirm that pathogenic Leptospira species circulate among agro-pastoralists living in Katavi-Rukwa ecosystem
title_fullStr Human leptospirosis in Tanzania: sequencing and phylogenetic analysis confirm that pathogenic Leptospira species circulate among agro-pastoralists living in Katavi-Rukwa ecosystem
title_full_unstemmed Human leptospirosis in Tanzania: sequencing and phylogenetic analysis confirm that pathogenic Leptospira species circulate among agro-pastoralists living in Katavi-Rukwa ecosystem
title_short Human leptospirosis in Tanzania: sequencing and phylogenetic analysis confirm that pathogenic Leptospira species circulate among agro-pastoralists living in Katavi-Rukwa ecosystem
title_sort human leptospirosis in tanzania: sequencing and phylogenetic analysis confirm that pathogenic leptospira species circulate among agro-pastoralists living in katavi-rukwa ecosystem
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4902944/
https://www.ncbi.nlm.nih.gov/pubmed/27287703
http://dx.doi.org/10.1186/s12879-016-1588-x
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