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      Molecular evidence for the presence of Leptospira borgpetersenii in synanthropic rodents in the Nautla region, Veracruz, México

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          Abstract

          Abstract: The genus Leptospira encompasses ten species of spirochetes capable of infecting mammals, particularly rodents. In México, studies focused on the detection of Leptospira sp. in rodents are scarce, all of them restricted to three states of the Gulf of México. For this reason, this work aimed to identify the diversity of Leptospira species associated with synanthropic rodents in Veracruz, a state where leptospirosis is endemic. Rodents were sampled with Sherman traps placed in 10 Production Units across the Nautla region. Animals were euthanized and their kidneys removed. Subsequently, a 474-bp segment of the outer membrane protein LipL32, present in all pathogenic species, was amplified and sequenced. Sequences were compared vs. reference using the BLAST algorithm: a phylogenetic reconstruction was carried out using the Maximum Likelihood method. In addition, the prevalence of infection in each Production Unit was estimated. Twenty eight rodents of a single species (Mus musculus) were caught. Leptospira DNA was detected in 17 samples (62.9 %, CI95 % 42.3 to 80.6) from seven localities in the Nautla region. The sequences recovered exhibited 99-100% identity to each other and 99 % identity with Leptospira borgspetersenii sequences deposited in GenBank. This study confirms the presence of L. borgspetersenii in rodents, particularly in M. musculus, in México. This study increases the inventory of pathogenic leptospires for the state of Veracruz to three species.

          Translated abstract

          Resumen: El género Leptospira engloba 10 especies de espiroquetas capaces de infectar mamíferos, particularmente roedores. En México se han realizado escasos estudios para la detección de Leptospira sp. en roedores, todos ellos restringidos a tres estados del Golfo de México. Por tal motivo el objetivo del presente trabajo fue identificar la diversidad de leptospiras en roedores sinantrópicas de Veracruz, un estado endémico de leptospirosis. Para la colecta de roedores, se colocaron trampas tipo Sherman en 10 unidades de producción de la región Nautla. Los animales se sacrificaron y se obtuvieron los riñones. Posteriormente se amplificó y secuenció un segmento de 474 pb de la proteína exterior de membrana LipL32 presente en las leptospiras patógenas. Posteriormente se compararon las secuencias con las de referencia mediante el uso del algoritmo BLAST y se realizó una reconstrucción filogenética mediante el método de Máxima Verosimilitud. Adicionalmente se obtuvieron las prevalencias de la infección por unidad de producción. Se colectaron 28 roedores de una única especie (Mus musculus). Se detectó la presencia de ADN de Leptospira en 17 muestras (62.9 %; IC95 % 42.3 a 80.6) procedentes de siete localidades de la región Nautla. Las secuencias recuperadas exhibieron una similitud del 99-100 % entre si y una identidad del 99 % con secuencias de referencia de Leptospira borgspetersenii depositadas en GenBank. Este estudio confirma la presencia de L. borgspetersenii en roedores y en particular con M. musculus en México. Este estudio incrementa a tres especies el inventario de leptospiras patógenas para el estado de Veracruz.

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          Most cited references21

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          MEGA6: Molecular Evolutionary Genetics Analysis version 6.0.

          We announce the release of an advanced version of the Molecular Evolutionary Genetics Analysis (MEGA) software, which currently contains facilities for building sequence alignments, inferring phylogenetic histories, and conducting molecular evolutionary analysis. In version 6.0, MEGA now enables the inference of timetrees, as it implements the RelTime method for estimating divergence times for all branching points in a phylogeny. A new Timetree Wizard in MEGA6 facilitates this timetree inference by providing a graphical user interface (GUI) to specify the phylogeny and calibration constraints step-by-step. This version also contains enhanced algorithms to search for the optimal trees under evolutionary criteria and implements a more advanced memory management that can double the size of sequence data sets to which MEGA can be applied. Both GUI and command-line versions of MEGA6 can be downloaded from www.megasoftware.net free of charge.
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            Biodiversity of Environmental Leptospira : Improving Identification and Revisiting the Diagnosis

            Leptospirosis is an important environmental disease and a major threat to human health causing at least 1 million clinical infections annually. There has recently been a growing interest in understanding the environmental lifestyle of Leptospira. However, Leptospira isolation from complex environmental samples is difficult and time-consuming and few tools are available to identify Leptospira isolates at the species level. Here, we propose a polyphasic isolation and identification scheme, which might prove useful to recover and identify environmental isolates and select those to be submitted to whole-genome sequencing. Using this approach, we recently described 12 novel Leptospira species for which we propose names. We also show that MALDI-ToF MS allows rapid and reliable identification and provide an extensive database of Leptospira MALDI-ToF mass spectra, which will be valuable to researchers in the leptospirosis community for species identification. Lastly, we also re-evaluate some of the current techniques for the molecular diagnosis of leptospirosis taking into account the extensive and recently revealed biodiversity of Leptospira in the environment. In conclusion, we describe our method for isolating Leptospira from the environment, confirm the usefulness of mass spectrometry for species identification and propose names for 12 novel species. This also offers the opportunity to refine current molecular diagnostic tools.
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              Systematics of leptospiraceae.

              Leptospires are spirochetes that may be free-living saprophytes found in freshwater or may cause acute or chronic infection of animals. The family Leptospiraceae comprises three genera: Leptospira Leptospira Leptonema Leptonema, and Turneriella Turneriella. Within the genus Leptospira, three clades can be distinguished, of pathogens, nonpathogens, and an intermediate group. Leptospires are further divided into serovars; antigenically related serovars are clustered into serogroups for convenience.
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                Author and article information

                Contributors
                Role: ND
                Role: ND
                Role: ND
                Role: ND
                Role: ND
                Role: ND
                Role: ND
                Role: ND
                Journal
                therya
                Therya
                Therya
                Asociación Mexicana de Mastozoología A. C. (La Paz, Baja California Sur, Mexico )
                2007-3364
                August 2019
                : 10
                : 2
                : 171-174
                Affiliations
                [2] orgnameUniversidad Veracruzana orgdiv1Facultad de Ciencias Biológicas y Agropecuarias Mexico rserna@ 123456uv.mx
                [3] Ciudad de México orgnameUniversidad Nacional Autónoma de México orgdiv1Facultad de Medicina Mexico sok10108@ 123456gmail.com
                [1] orgnameUniversidad Veracruzana orgdiv1Facultad de Medicina Veterinaria y Zootecnia Mexico iori_luna25@ 123456hotmail.com
                Article
                S2007-33642019000200171
                10.12933/therya-19-723
                9fdc50a4-a34b-4ea6-84ae-792e161a22dc

                This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

                History
                : 27 November 2018
                : 21 March 2019
                Page count
                Figures: 0, Tables: 0, Equations: 0, References: 21, Pages: 4
                Product

                SciELO Mexico

                Categories
                Notes

                Mexico,pathogens,small mammals,spirochetes
                Mexico, pathogens, small mammals, spirochetes

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