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Table of Contents
SHORT RESEARCH COMMUNICATION
Year : 2021  |  Volume : 58  |  Issue : 2  |  Page : 175-177

Detection of Leishmania tarentolae DNA in Sergentomyia antennata in Togo


1 Université de Reims Champagne-Ardenne, Faculté de Pharmacie, ANSES, SFR Cap Santé, EA7510 ESCAPE - USC VECPAR, 51 rue Cognacq-Jay, 51096 Reims cedex, France
2 National Reference Center for Leishmaniases, CHU (University Hospital Centre) of Montpellier & University Montpellier 1 (Faculty of Medicine), Laboratoire de Parasitologie-Mycologie, Montpellier, France et CNRS 5290- IRD 224-University Montpellier 1, Research Unit “MIVEGEC”, Montpellier, France
3 Université de Reims Champagne-Ardenne, Faculté de Pharmacie, ANSES, SFR Cap Santé, EA7510 ESCAPE - USC VECPAR, 51 rue Cognacq-Jay, 51096 Reims cedex, France Laboratoire de Parasitologie-Mycologie, CHU de Reims, 51100 Reims, France

Date of Submission16-May-2020
Date of Acceptance25-Sep-2020
Date of Web Publication13-Jan-2022

Correspondence Address:
Prof. Jérôme Depaquit
Université de Reims Champagne-Ardenne, Faculté de Pharmacie, ANSES, SFR Cap Santé, EA7510 ESCAPE – USC VECPAR, 51 rue Cognacq-Jay, 51096 Reims cedex, France Laboratoire de Parasitologie- Mycologie, CHU de Reims, 51100 Reims
France
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/0972-9062.316270

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  Abstract 


Keywords: Sergentomyia antennata; Leishmania tarentolae; Togo; molecular epidemiology


How to cite this article:
Ferlet E, Martinet JP, Randrianambinintsoa FJ, Ravel C, Depaquit J. Detection of Leishmania tarentolae DNA in Sergentomyia antennata in Togo. J Vector Borne Dis 2021;58:175-7

How to cite this URL:
Ferlet E, Martinet JP, Randrianambinintsoa FJ, Ravel C, Depaquit J. Detection of Leishmania tarentolae DNA in Sergentomyia antennata in Togo. J Vector Borne Dis [serial online] 2021 [cited 2022 Jan 26];58:175-7. Available from: https://www.jvbd.org/text.asp?2021/58/2/175/316270

According to the World Health Organization, cutaneous leishmaniasis (CL) is endemic in all the west African countries except Sierra Leone, Liberia, Benin and Togo[1]. Visceral leishmaniasis (VL) is considered endemic in Senegal, Mauritania, Niger and Côte d’Ivoire and cases were previously reported in Nigeria. Togo is considered as a country free of leishmaniasis despite the report of a single case of VL in Lama-Kara (northern region)[2]. Consequently, the fauna of Phlebotomine sandflies of Togo including 17 species remains poorly documented in this country[3],[4],[5]. In the present study, we caught Phlebotomine sandflies and did a Leishmania DNA screening on the females.

Two CDC miniature light traps have been used for this study carried out in 2017 from July 2 to August 3. They have been put in the following localities: Lomé (4 traps/ nights), Adétikopé (Lomé's suburb: 4 trap/night), Goumoukopé (4 traps/nights), Edoh-Condji (2 traps/nights), Glidji (2 traps/nights), Fiata (4 traps/nights), Atoeta (2 traps/nights), Melly-Djigbé (2 traps/nights), and Kpalimé (4 traps/nights). The traps were left overnight, from 5 pm to 8 am the following morning. Traps were placed in gardens, courtyards, near fields, near dogs and henhouses, palms, in anthropised places. Sandflies were stored in 96% ethanol.

The head, thorax and genitalia were cut off in a drop of ethanol. Soft tissues were lysed in a bath of KOH 10%, then bleached in Marc-André solution, and mounted between microscope slide and cover slide in Euparal® for species identification after dehydration in successive alcoholic baths. Measurements and counts were made by using the Stream Motion software (Olympus, Japan) and a video camera connected to the microscope.

The abdomen related to the specimen was stored dried in a vial at -20°C before DNA extraction. DNA was extracted using the QIAmp®DNA Mini Kit (Qiagen, Germany) with a final elution volume of 150 μL. Detection of trypanosomatid DNA was carried out by PCR. We selected primers hybridizing all Leishmania species as well as Endotrypanum monterrogeii[6]. We amplified a 120 bp kDNA corresponding to the conserved region of kinetoplast minicircles. We used Leishmania major and L. tropica DNA samples as positive controls and included a negative control.

To confirm the identity of a detected kinetoplastid, we also amplified a part of the 18S ribosomal DNA using a LightCycler® 480 SYBR Green I Master - Roche Life Science thanks to primers SD (3′-TTAATTTGACTCAACACG-5′) and SR (3′-CGACGGGCGGTGTGTACA-5′) according to the following PCR cycles: initial denaturation of 5′ (95°C) then 40 cycles (56°C for 30”/72°C for 30”/95°C for 30”). The sequencing of the amplicons has been done by Sanger’s technique in both directions.

A total of 29 Phlebotomine sandflies has been caught. Their identifications are indicated in [Table 1]. Out of 17 females tested by PCR for the presence of trypanosomatid kDNA, one identified as Se. antennata was positive. BLAST (www.blast.ncbi.nlm.nih.gov) analysis of the 120 bp PCR product revealed a full homology with the Leishmania tarentolae sequence AF380748. The partial rDNA sequences are 100% homologous with those deposited in GenBank under the accession numbers M84225 and X53916. The SSU rDNA sequence obtained in the present study has been deposited in Genbank under accession number MK697739.
Table 1: Sandflies species sampling

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Visceral or cutaneous cases of leishmaniasis were reported in the Togo neighboring countries, except Benin. For example, in Burkina Faso, 1845 CL cases were reported the town of Ouagadoudou between 1996 and 1998[7]. In Ghana, an active case search found 2348 infected individuals in the Ho district in 2002 and over 6,000 cases in 2003[9]. In Nigeria, a study in Kaena, in the Plateau state, found 3.9% of the inhabitants had CL[10]. The risk of acquiring CL has been reported to increase considerably with human activity and epidemics of CL have been associated with deforestation, road construction, wars, or other activities where humans intrude the habitat of the vector[11]. In spite of this, little is currently known regarding the epidemiology of cutaneous and visceral leishmaniasis in west African counties. Except a significant problem of under-reporting there is no plausible hypotheses to explain the absence of described cases in Togo or Benin who are surrounded by endemic countries. There is no evidence of reservoir hosts or anthropophilic sand flies.

The sampling of the present study is limited. It includes species which have previously been recorded in the country. To date, the 17 species have been recorded in Togo: Phlebotomus duboscqi, Ph. rodhaini, Grassomyia ghesqueirei, Gr. squamipleuris, Sergentomyia adleri, Se. affinis vorax, Se. africana, Se. antennata, Se. bedfordi, Se. buxtoni, Se. clydei, Se. freetownensis, Se. ingrami, Se. magna, Se. schoutedeni, Se. schwetzi, and Se. simillima[3],[4],[5].

Regarding the detection of Leishmania DNA, there are a few records of the isolation of Leishmania or the detection of Leishmania DNA in Sergentomyia spp.[12],[13]. In the present study, we detected the carrying of L. tarentolae by a female Se. antennata. This result is not surprising because the genus Sergentomyia is considered as feeding primarily on cold-blooded animals such as lizards[3]. Leishmania tarentolae is mainly studied as a laboratory model for gene expression but little is known about its distribution and vectors. Some Phlebotomine sandflies have been found infected by L. tarentolae or carrying Leishmania DNA. In Italy, Se. minuta has been found naturally infected[14] or harbouring DNA. L. tarentolae-like DNA has also been detected in Se. minuta in Portugal. More recently, L. tarentolae DNA has also been detected in Se. minuta from Spain and Iraq[15],[16],[17]. In the past, some promastigotes have been isolated from Sergentomyia spp. without typing. To our knowledge, this parasite has never been detected in Se. antennata. The detection of L. tarentolae DNA in this sandfly does not mean that it acts as a vector. However, it is now documented that some Sergentomyia can feed on mammals and their vectorial role could not be neglected[12]. In southeastern Asia, an area which has long been considered to be free of leishmaniasis, DNA of Leishmania martiniquensis, a species causing an emerging disease in man, has been detected in some Sergentomyia spp.[18] despite their role in the transmission of these Leishmania remains unclear and stands as one of the most exciting challenges in the field of the epidemiology of leishmaniases. In Ghana, a species closely related to L. martiniquensis causing CL has been recently described[19] and Sergentomyia spp. could be considered as candidates to its transmission.

Conflict of interest: None



 
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Maroli M, Gramiccia M, Gradoni L, Ready PD, Smith DF, Aquino C. Natural infections of phlebotomine sandflies with Trypanosomatidae in Central and South Italy. Trans R Soc Trop Med Hyg 1988; 82: 227–8.  Back to cited text no. 14
    
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