Publications by authors named "Zoubir Harrat"

25 Publications

  • Page 1 of 1

Dipeptidyl peptidase III as a DNA marker to investigate epidemiology and taxonomy of Old World Leishmania species.

PLoS Negl Trop Dis 2021 Jul 26;15(7):e0009530. Epub 2021 Jul 26.

Laboratory of Molecular Epidemiology and Experimental Pathology, Institut Pasteur de Tunis, Université de Tunis El Manar, Tunisia.

Background: Dipeptidyl peptidase III (DPPIII) member of M49 peptidase family is a zinc-dependent metallopeptidase that cleaves dipeptides sequentially from the N-terminus of its substrates. In Leishmania, DPPIII, was reported with other peptidases to play a significant role in parasites' growth and survival. In a previous study, we used a coding sequence annotated as DPPIII to develop and evaluate a PCR assay that is specific to dermotropic Old World (OW) Leishmania species. Thus, our objective was to further assess use of this gene for Leishmania species identification and for phylogeny, and thus for diagnostic and molecular epidemiology studies of Old World Leishmania species.

Methodology: Orthologous DDPIII genes were searched in all Leishmania genomes and aligned to design PCR primers and identify relevant restriction enzymes. A PCR assays was developed and seventy-two Leishmania fragment sequences were analyzed using MEGA X genetics software to infer evolution and phylogenetic relationships of studied species and strains. A PCR-RFLP scheme was also designed and tested on 58 OW Leishmania strains belonging to 8 Leishmania species and evaluated on 75 human clinical skin samples.

Findings: Sequence analysis showed 478 variable sites (302 being parsimony informative). Test of natural selection (dN-dS) (-0.164, SE = 0.013) inferred a negative selection, characteristic of essential genes, corroborating the DPPIII importance for parasite survival. Inter- and intra-specific genetic diversity was used to develop universal amplification of a 662bp fragment. Sequence analyses and phylogenies confirmed occurrence of 6 clusters congruent to L. major, L. tropica, L. aethiopica, L. arabica, L. turanica, L. tarentolae species, and one to the L. infantum and L. donovani species complex. A PCR-RFLP algorithm for Leishmania species identification was designed using double digestions with HaeIII and KpnI and with SacI and PvuII endonucleases. Overall, this PCR-RFLP yielded distinct profiles for each of the species L. major, L. tropica, L. aethiopica, L. arabica and L. turanica and the L. (Sauroleishmania) L. tarentolae. The species L. donovani, and L. infantum shared the same profile except for strains of Indian origin. When tested on clinical samples, the DPPIII PCR showed sensitivities of 82.22% when compared to direct examination and was able to identify 84.78% of the positive samples.

Conclusion: The study demonstrates that DPPIII gene is suitable to detect and identify Leishmania species and to complement other molecular methods for leishmaniases diagnosis and epidemiology. Thus, it can contribute to evidence-based disease control and surveillance.
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http://dx.doi.org/10.1371/journal.pntd.0009530DOI Listing
July 2021

Cutaneous Leishmaniasis in Algeria; Highlight on the Focus of M'Sila.

Microorganisms 2021 Apr 29;9(5). Epub 2021 Apr 29.

Laboratoire d'Eco-épidémiologie Parasitaire et Génétique des Populations, Route du Petit Staoueli, Institut Pasteur d'Algérie, Dely-Brahim 16047, Algeria.

Algeria ranks second after Afghanistan for the incidence of cutaneous leishmaniasis (CL) worldwide. Here, we report a 34-years retrospective analysis of CL in Algeria and focused on the most affected region, the M'Sila province. All 66 cutaneous isolates corresponded to (.) . Our study of the sandfly and rodent fauna further highlighted the high density of and additional phlebotomine species of medical importance, not previously identified in M'Sila. Wild rodents belonging to nine species were trapped in M'Sila, and and were found infected by . In addition, was isolated from two visceral leishmaniasis cases, one dog and its proven vectors (, , and ) inventoried during the survey. The high incidence of CL in the M'Sila province is likely a consequence of the increase in minimum temperatures recorded that constitutes suitable conditions for establishing a high endemicity and leads to an explosive rise in leishmaniases cases in this region. A thorough investigation of the underlying risk factors is urgently needed to detect new cases earlier. All these would improve the preparedness to fight the disease.
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http://dx.doi.org/10.3390/microorganisms9050962DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8146893PMC
April 2021

New Haplotypes of Trypanosoma evansi Identified in Dromedary Camels from Algeria.

Acta Parasitol 2021 Mar 2;66(1):294-302. Epub 2021 Jan 2.

Laboratoire D'Eco-Épidémiologie Parasitaire Et Génétique Des Populations, Institut Pasteur D'Algérie, 01 Rue du Petit Staouéli Dely-Brahim, 16302, Algiers, Algeria.

Purpose: Surra is a zoonotic disease caused by Trypanosoma evansi (Trypanozoon), a salivary trypanosome native to Africa which affects a wide range of mammals worldwide and causes mortality and significant economic loss. The present study was devoted to the molecular characterization of T. evansi derived from naturally infected dromedary camels in Algeria.

Methods: A total of 148 blood samples were collected from mixed age camels living in one of four geographic regions (Ouargla, El Oued, Biskra and Ghardaia) of Algeria. Samples underwent PCR amplification and sequencing of the internal transcribed spacer 1 (ITS1) complete sequence.

Results: DNA of Trypanosoma spp. was found in 19 camels (12.84%). Trypanosoma spp. molecular positivity was not affected by sex (p = 0.50), age (p = 0.08), or geographic location (p = 0.12). Based on multiple sequence alignment of the obtained DNA sequences with representative T. evansi ITS1 sequences available globally, the Algerian sequences were grouped within four different haplotypes including two which were original.

Conclusion: Results of this study provide preliminary data on which future studies of genetic diversity and molecular epidemiology of T. evansi can be based.
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http://dx.doi.org/10.1007/s11686-020-00316-wDOI Listing
March 2021

First Detection of () (Diptera: Culicidae) in Algiers, the Capital City of Algeria.

J Arthropod Borne Dis 2019 Dec 31;13(4):420-425. Epub 2019 Dec 31.

Laboratoire d'Eco-épidémiologie Parasitaire et Génétique des Populations, Institut Pasteur d'Algérie, Route Petit Staouéli, Dely Ibrahim, Alger, Algérie.

Background: Based on the reporting of the presence of stripped mosquitoes by a citizen in the Algiers residential neighborhood of Bir-Khadem, where residents experienced huge daytime mosquito nuisance an entomological investigation was carried out in July 2016.

Methods: Ovitraps and BG sentinel traps baited with Lure were used during three consecutive days to collect adult mosquitoes. Eighteen residential houses of the Bir-Khadem neighborhood were also inspected to search larvae breeding sites such as water fountains, baskets and flowerpots.

Results: A total of 57 specimens were collected in five villas, consisting of 21 eggs, 20 larvae and 16 adults.

Conclusion: This is the first record of this invasive species in Algiers.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7188769PMC
December 2019

Coxiella burnetii in camels (Camelus dromedarius) from Algeria: Seroprevalence, molecular characterization, and ticks (Acari: Ixodidae) vectors.

Acta Trop 2020 Jun 13;206:105443. Epub 2020 Mar 13.

Laboratoire d'Eco-épidémiologie Parasitaire et Génétique des Populations, Institut Pasteur d'Algérie, 16000 Algiers, Algeria. Electronic address:

Q fever is a widespread zoonotic disease caused by Coxiella burnetii that most commonly infects not only a variety of mammals but also arthropods and in particularly ticks. The aim of this study was to detect C. burnetii infection in camels including ixodid ticks using serological and molecular assays. Between July 2018 to June 2019, blood samples from 184 male and female camels (Camelus dromedarius) were collected from 3 regions of South-East Algeria and serum samples were tested for antibodies against Coxiella burnetii using indirect enzyme-linked immunosorbent assay (ELISA) kit. The positive sera and a total of 60 ticks were tested by quantitative PCR (qPCR) for detection of C. burnetii with primers and probes specific to the transposon-like repetitive region (IS1111 gene). Positive samples were genotyped by amplification and sequencing of partial sequences based on the IS1111 gene. The seroprevalence of antibodies against C. burnetii was 75.5%. Statistical analysis pointed out three potential risk factors associated with Q fever infection: geographic location, age class and season. No positive DNA of camel blood sample was observed. However, five Hyalomma dromedarii, one H. impeltatum and one H. excavatum tick species were detected positive for Coxiella burnetii DNA by qPCR, with an overall prevalence rate of 11.66% (7/60). The revealed Algerian strains by phylogenetic and comparative analysis of the IS1111 nucleotide sequences were clustered with several pathogenic C. burnetii strains isolated from ticks, human, and cattle located in Tunisia, Greece and in some Mediterranean countries, respectively. The study results clearly indicate that camels and their ticks in Algeria may play an important role as a reservoir for C. burnetii and can be considered as a significant source of Q fever transmission to other animal species and humans.
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http://dx.doi.org/10.1016/j.actatropica.2020.105443DOI Listing
June 2020

Towards harmonisation of entomological surveillance in the Mediterranean area.

PLoS Negl Trop Dis 2019 06 13;13(6):e0007314. Epub 2019 Jun 13.

French National Research Institute for Sustainable Development, Research unit MIVEGC IRD-CNRS-Montpellier University, Montpellier, France.

Background: The Mediterranean Basin is historically a hotspot for trade, transport, and migration. As a result, countries surrounding the Mediterranean Sea share common public health threats. Among them are vector-borne diseases, and in particular, mosquito-borne viral diseases are prime candidates as (re)emerging diseases and are likely to spread across the area. Improving preparedness and response capacities to these threats at the regional level is therefore a major issue. The implementation of entomological surveillance is, in particular, of utmost importance. Guidance in designing entomological surveillance systems is critical, and these systems may pursue different specific objectives depending on the disease. The purpose of the proposed review is to draw up guidelines for designing effective and sustainable entomological surveillance systems in order to improve preparedness and response. However, we make it clear that there is no universal surveillance system, so the thinking behind harmonisation is to define evidence-based standards in order to promote best practises, identify the most appropriate surveillance activities, and optimise the use of resources. Such guidance is aimed at policymakers and diverse stakeholders and is intended to be used as a framework for the implementation of entomological surveillance programmes. It will also be useful to collaborate and share information with health professionals involved in other areas of disease surveillance. Medical entomologists and vector control professionals will be able to refer to this report to advocate for tailored entomological surveillance strategies. The main threats targeted in this review are the vectors of dengue virus, chikungunya virus, Zika virus, West Nile virus, and Rift Valley fever virus. The vectors of all these arboviruses are mosquitoes.

Methods: Current knowledge on vector surveillance in the Mediterranean area is reviewed. The analysis was carried out by a collaboration of the medical entomology experts in the region, all of whom belong to the MediLabSecure network, which is currently funded by the European Union and represents an international effort encompassing 19 countries in the Mediterranean and Black Sea region.

Findings: Robust surveillance systems are required to address the globalisation of emerging arboviruses. The prevention and management of mosquito-borne viral diseases must be addressed in the prism of a One Health strategy that includes entomological surveillance as an integral part of the policy. Entomological surveillance systems should be designed according to the entomological and epidemiological context and must have well-defined objectives in order to effect a tailored and graduated response. We therefore rely on different scenarios according to different entomological and epidemiological contexts and set out detailed objectives of surveillance. The development of multidisciplinary networks involving both academics and public authorities will provide resources to address these health challenges by promoting good practises in surveillance (identification of surveillance aims, design of surveillance systems, data collection, dissemination of surveillance results, evaluation of surveillance activities) and through the sharing of effective knowledge and information. These networks will also contribute to capacity building and stronger collaborations between sectors at both the local and regional levels. Finally, concrete guidance is offered on the vector of the main arbovirus based on the current situation in the area.
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http://dx.doi.org/10.1371/journal.pntd.0007314DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6563966PMC
June 2019

Meta-analysis and discussion on challenges to translate Leishmania drug resistance phenotyping into the clinic.

Acta Trop 2019 Mar 9;191:204-211. Epub 2019 Jan 9.

Laboratory of Eco-epidemiology and Parasitic Population Genetics, Pasteur Institute of Algier, Algier, Algeria. Electronic address:

Antimicrobial resistance (AMR) threatens the prevention and treatment of infections caused by a large range of microorganisms. Leishmania is not an exception and treatment failure due to drug-resistant organisms is increasingly reported. Currently, no molecular methods and marker are validated to track drug-resistant organism and antimicrobial susceptibility tests are roughly not amenable to a clinical setting. Taking these facts into account, it is essential to reflect on ways to translate basic knowledge into methodologies aimed to diagnose leishmania drug resistance. As a matter of fact, a meta-analysis of the literature discloses the reliability of the promastigotes antimicrobial susceptibility tests (AST) to predict intracellular amastigotes susceptibility status. Promastigote cultures that are easy to perform, typically inexpensive and amenable to standardization should represent a candidate to diagnose resistance. Using AST performed on promastigote, we propose a way to improve leishmania drug resistance diagnosis in the framework of guidance and guideline of the bacterial drug resistance diagnosis. In this review, we highlight challenges that remained and discuss the definition of clinical breakpoints, including the epidemiological cutoff (ECOFF), to track drug-resistant isolates. Our analysis paves the ways to standardize and analyze anti-leishmania susceptibility tests output in order to guide the characterization of drug-resistant isolates, the clinical decision during treatment and the search for new molecular markers.
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http://dx.doi.org/10.1016/j.actatropica.2019.01.009DOI Listing
March 2019

Genome Dynamics during Environmental Adaptation Reveal Strain-Specific Differences in Gene Copy Number Variation, Karyotype Instability, and Telomeric Amplification.

mBio 2018 11 6;9(6). Epub 2018 Nov 6.

Unité de Parasitologiemoléculaire et Signalisation, Institut Pasteur, Paris, France

Protozoan parasites of the genus adapt to environmental change through chromosome and gene copy number variations. Only little is known about external or intrinsic factors that govern genomic adaptation. Here, by conducting longitudinal genome analyses of 10 new clinical isolates, we uncovered important differences in gene copy number among genetically highly related strains and revealed gain and loss of gene copies as potential drivers of long-term environmental adaptation in the field. In contrast, chromosome rather than gene amplification was associated with short-term environmental adaptation to culture. Karyotypic solutions were highly reproducible but unique for a given strain, suggesting that chromosome amplification is under positive selection and dependent on species- and strain-specific intrinsic factors. We revealed a progressive increase in read depth towards the chromosome ends for various isolates, which may represent a nonclassical mechanism of telomere maintenance that can preserve integrity of chromosome ends during selection for fast growth. Together our data draw a complex picture of genomic adaptation in the field and in culture, which is driven by a combination of intrinsic genetic factors that generate strain-specific phenotypic variations, which are under environmental selection and allow for fitness gain. Protozoan parasites of the genus cause severe human and veterinary diseases worldwide, termed leishmaniases. A hallmark of biology is its capacity to adapt to a variety of unpredictable fluctuations inside its human host, notably pharmacological interventions, thus, causing drug resistance. Here we investigated mechanisms of environmental adaptation using a comparative genomics approach by sequencing 10 new clinical isolates of the , , and complexes that were sampled across eight distinct geographical regions. Our data provide new evidence that parasites adapt to environmental change in the field and in culture through a combination of chromosome and gene amplification that likely causes phenotypic variation and drives parasite fitness gains in response to environmental constraints. This novel form of gene expression regulation through genomic change compensates for the absence of classical transcriptional control in these early-branching eukaryotes and opens new venues for biomarker discovery.
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http://dx.doi.org/10.1128/mBio.01399-18DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6222132PMC
November 2018

Identification of mosquitoes (Diptera: Culicidae): an external quality assessment of medical entomology laboratories in the MediLabSecure Network.

Parasit Vectors 2018 Oct 23;11(1):553. Epub 2018 Oct 23.

French National Research Institute for Sustainable Development, Research Unit MiVEGEC, IRD-CNRS-Montpellier University, Montpellier, France.

Background: Identification of vectors is of prime importance in the field of medical entomology for both operational and research purposes. An external quality assessment of mosquito identification capacities was carried out within the MediLabSecure Network, which is composed of laboratories located in 19 countries close to the European Union around the Mediterranean and Black seas.

Methods: A set of blind samples consisting of 7 or 8 adult mosquitoes and 4 larvae was given to each participant laboratory. In all, 138 adult mosquitoes and 76 larvae of different species were distributed for genus and species identification.

Results: All identifications were exclusively morphology based. Overall, 81% of identifications were correct at the genus level, 64% at the species level. The results were highly varied among the 19 participating laboratories. The levels of correct identifications were: 100% (three laboratories), 90-95% (four laboratories), 50-75% (six laboratories) and < 50% (six laboratories).

Conclusions: This evaluation showed the need to maintain efforts in capacity building and quality control in the field of medical entomology and, more specifically, in the morphological identification of the Culicidae.
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http://dx.doi.org/10.1186/s13071-018-3127-7DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6199703PMC
October 2018

Antimony susceptibility of Leishmania isolates collected over a 30-year period in Algeria.

PLoS Negl Trop Dis 2018 03 21;12(3):e0006310. Epub 2018 Mar 21.

IRD, Univ Montpellier, MIVEGEC, Montpellier, France.

Background: In Algeria, the treatment of visceral and cutaneous leishmanioses (VL and CL) has been and continues to be based on antimony-containing drugs. It is suspected that high drug selective pressure might favor the emergence of chemoresistant parasites. Although treatment failure is frequently reported during antimonial therapy of both CL and VL, antimonial resistance has never been thoroughly investigated in Algeria. Determining the level of antimonial susceptibility, amongst Leishmania transmitted in Algeria, is of great importance for the development of public health policies.

Methodology/principal Findings: Within the framework of the knowledge about the epidemiology of VL and CL amassed during the last 30 years, we sampled Leishmania isolates to determine their susceptibility to antimony. We analyzed a total of 106 isolates including 88 isolates collected between 1976 and 2013 in Algeria from humans, dogs, rodents, and phlebotomines and 18 collected from dogs in France. All the Algerian isolates were collected in 14 localities where leishmaniasis is endemic. The 50% inhibitory concentrations (IC50) of potassium antimony tartrate (the trivalent form of antimony, Sb(III)) and sodium stibogluconate (the pentavalent form of antimony, Sb(V)) were determined in promastigotes and intramacrophage amastigotes, respectively. The epidemiological cutoff (ECOFF) that allowed us to differentiate between Leishmania species causing cutaneous or visceral leishmaniases that were susceptible (S+) or insusceptible (S-) to the trivalent form of antimony was determined. The computed IC50 cutoff values were 23.83 μg/mL and 15.91 μg/mL for VL and CL, respectively. We report a trend of increasing antimony susceptibility in VL isolates during the 30-year period. In contrast, an increase in the frequency of S- phenotypes in isolates causing CL was observed during the same period. In our study, the emergence of S- phenotypes correlates with the inclusion of L. killicki (syn: L. tropica) isolates that cause cutaneous leishmaniasis and that have emerged in Algeria during the last decade.

Conclusion/significance: Our results provide insight into the spatiotemporal dynamics of Leishmania antimony susceptibility in Algeria. We highlight the need for the future implementation of an effective methodology to determine the antimony susceptibility status of Leishmania isolates to detect the emergence of and prevent the dissemination of drug-resistant strains.
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http://dx.doi.org/10.1371/journal.pntd.0006310DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5889277PMC
March 2018

Antimonial susceptibility and in vivo behaviour of Leishmania major isolates collected in Algeria before and after treatment.

Acta Trop 2018 Apr 24;180:7-11. Epub 2017 Dec 24.

IRD, Univ. Montpellier, InterTryp, Montpellier, France; IRD, Univ. Montpellier, MiVegec, Montpellier, France. Electronic address:

The repercussions of cutaneous leishmaniasis therapy on the behaviour and drug susceptibility of Leishmania major parasites is poorly documented. This study explored the link between antimonial susceptibility and in vivo behaviour in Leishmania major isolates collected before and after treatment in Algeria. This study was performed on 3 isolates collected from patients prior to treatment and paired with 3 isolates collected from the same patient after treatment failure. Their in vitro susceptibility towards trivalent (SbIII) and pentavalent (SbV) antimony were ascertained, and their in vivo behaviour was evaluated by determining their capacity to disseminate, proliferate and induce lesions in mice. No relationship was observed between in vitro antimony resistance and parasite fitness in the murine model.
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http://dx.doi.org/10.1016/j.actatropica.2017.12.020DOI Listing
April 2018

Isolation and characterization of an anti-leishmanial disintegrin from Cerastes cerastes venom.

J Biochem Mol Toxicol 2018 Feb 26;32(2). Epub 2017 Dec 26.

USTHB, Faculty of Biological Sciences, Laboratory of Cellular and Molecular Biology, Bab Ezzouar, Algiers, 16111, Algeria.

Investigating new antimicrobial and antiparasitic components from Viperidae venoms represents an alternative therapeutic strategy. In this study, we report the characterization of a disintegrin isolated from Cerastes cerastes venom, exhibiting antiparasitic activity on Leishmania infantum promastigotes. Indeed, isolated disintegrin, referred to Disintegrin_Cc, induced 84.75% of parasiticidal activity and deep morphological alterations on the parasites. SDS-PAGE analysis indicated that this disintegrin was homogenous. This dimeric disintegrin of 14,193.97 Da contains an RGD domain and four intramolecular disulfide bridges. It presents a high percentage of identity with other related snake disintegrins. Predicted 3D structure indicated that this peptide shares partial homology with well-known active antimicrobial peptides. Disintegrin_Cc inhibited 80% of arachidonic acid-induced platelet aggregation. The obtained results suggest that the isolated molecule plays a dual role as a disintegrin and as an anti-leishmanial compound. This component could be useful as a drug in the treatment of leishmaniasis.
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http://dx.doi.org/10.1002/jbt.22018DOI Listing
February 2018

Presence of Atypical Form in Algeria.

J Arthropod Borne Dis 2017 Mar 14;11(1):139-146. Epub 2017 Mar 14.

Laboratoire d'Eco-Epidémiologie Parasitaire et Génétique des Populations, Institut Pasteur d'Algérie, Algeria.

Background: and are two phlebotomine sand fly species morphologically similar and differing in males only by the shape of the copulatory valves which are bifurcated in , tip long and tapered in .

Methods: A count of the median coxite setae was carried out on 208 specimens from the collections of Dedet and of Parrot, identified previously as and on 38 male sand flies captured during the year 2012-2013, in order to seek the presence of atypical form.

Results: The analysis revealed the presence of 33/246 (13%) atypical previously confused with species and whose distribution is mainly located in the semi-arid and arid bioclimatic regions.

Conclusion: This study proved for the first time the presence of atypical form of in Algeria.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5629296PMC
March 2017

Identification of Leishmania by Matrix-Assisted Laser Desorption Ionization-Time of Flight (MALDI-TOF) Mass Spectrometry Using a Free Web-Based Application and a Dedicated Mass-Spectral Library.

J Clin Microbiol 2017 10 19;55(10):2924-2933. Epub 2017 Jul 19.

Laboratoire de Parasitologie-Mycologie, CHU Timone, Université d'Aix-Marseille, Marseille, France

Human leishmaniases are widespread diseases with different clinical forms caused by about 20 species within the genus. species identification is relevant for therapeutic management and prognosis, especially for cutaneous and mucocutaneous forms. Several methods are available to identify species from culture, but they have not been standardized for the majority of the currently described species, with the exception of multilocus enzyme electrophoresis. Moreover, these techniques are expensive, time-consuming, and not available in all laboratories. Within the last decade, mass spectrometry (MS) has been adapted for the identification of microorganisms, including However, no commercial reference mass-spectral database is available. In this study, a reference mass-spectral library (MSL) for isolates, accessible through a free Web-based application (mass-spectral identification [MSI]), was constructed and tested. It includes mass-spectral data for 33 different species, including species that infect humans, animals, and phlebotomine vectors. Four laboratories on two continents evaluated the performance of MSI using 268 samples, 231 of which were strains. All strains, but one, were correctly identified at least to the complex level. A risk of species misidentification within the , , and complexes was observed, as previously reported for other techniques. The tested application was reliable, with identification results being comparable to those obtained with reference methods but with a more favorable cost-efficiency ratio. This free online identification system relies on a scalable database and can be implemented directly in users' computers.
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http://dx.doi.org/10.1128/JCM.00845-17DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5625378PMC
October 2017

Surveillance of Arthropod-Borne Viruses and Their Vectors in the Mediterranean and Black Sea Regions Within the MediLabSecure Network.

Curr Trop Med Rep 2017 17;4(1):27-39. Epub 2017 Mar 17.

French National Research Institute for Sustainable Development, MIVEGEC Unit, IRD224-CNRS 5290-Montpellier University, Montpellier, France.

Purpose Of Review: Arboviruses, viruses transmitted by arthropods such as mosquitoes, ticks, sandflies, and fleas are a significant threat to public health because of their epidemic and zoonotic potential. The geographical distribution of mosquito-borne diseases such as West Nile (WN), Rift Valley fever (RVF), Dengue, Chikungunya, and Zika has expanded over the last decades. Countries of the Mediterranean and Black Sea regions are not spared. Outbreaks of WN are repeatedly reported in the Mediterranean basin. Human cases of RVF were reported at the southern borders of the Maghreb region. For this reason, establishing the basis for the research to understand the potential for the future emergence of these and other arboviruses and their expansion into new geographic areas became a public health priority. In this context, the European network "MediLabSecure" gathering laboratories in 19 non-EU countries from the Mediterranean and Black Sea regions seeks to improve the surveillance (of animals, humans, and vectors) by reinforcing capacity building and harmonizing national surveillance systems to address this important human and veterinary health issue. The aim of this review is to give an exhaustive overview of arboviruses and their vectors in the region.

Recent Findings: The data presented underline the importance of surveillance in the implementation of more adapted control strategies to combat vector-borne diseases. Partner laboratories within the MediLabSecure network present a wide range of infrastructures and have benefited from different training programs.

Summary: Although reporting of arboviral presence is not carried out in a systematic manner, the expansion of the area where arboviruses are present cannot be disputed. This reinforces the need for increasing surveillance capacity building in this region to prevent future emergences.
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http://dx.doi.org/10.1007/s40475-017-0101-yDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5362652PMC
March 2017

Development of a Murine Infection Model with Leishmania killicki, Responsible for Cutaneous Leishmaniosis in Algeria: Application in Pharmacology.

Biomed Res Int 2016 2;2016:7985104. Epub 2016 Feb 2.

Laboratoire d'Eco-Épidemiologie Parasitaire et Génétique des Populations, Institut Pasteur d'Algerie, Route de Petit Staouéli, Dely Brahim, Algiers, Algeria.

In Algeria, Leishmania infantum, Leishmania major, and Leishmania killicki (Leishmania tropica) are responsible for cutaneous leishmaniosis. We established a murine model of L. killicki infection to investigate its infective capacity, some immunophysiopathological aspects, and its suitability for pharmacological purposes. Following the injection of L. major or L. killicki metacyclic promastigotes in the ear dermis of BALB/c mice, the course of infection was followed. The infection with L. killicki caused slower lesion formation than with L. major. The presence of L. killicki or L. major DNA and parasites was detected in the ear dermis and in lymph nodes, spleen, and liver. Lesions induced by L. killicki were nonulcerative in their aspect, whereas those caused by L. major were highly ulcerative and necrotic, which matches well with the lesion phenotype reported in humans for L. killicki and L. major, respectively. The treatment of L. killicki lesions by injection of Glucantime® significantly reduced the lesion thickness and parasite burden. Ear dermal injection of BALB/c mice constitutes a model to study lesions physiopathology caused by L. killicki and presents interest for in vivo screening of new compounds against this pathogen, emerging in Algeria.
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http://dx.doi.org/10.1155/2016/7985104DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4754473PMC
November 2016

Comparison of Leishmania killicki (syn. L. tropica) and Leishmania tropica Population Structure in Maghreb by Microsatellite Typing.

PLoS Negl Trop Dis 2015 Dec 8;9(12):e0004204. Epub 2015 Dec 8.

Centre National de Référence des Leishmanioses, Département de Parasitologie-Mycologie, CHRU de Montpellier, Université de Montpellier, Montpellier, France.

Leishmania (L.) killicki (syn. L. tropica), which causes cutaneous leishmaniasis in Maghreb, was recently described in this region and identified as a subpopulation of L. tropica. The present genetic analysis was conducted to explore the spatio-temporal distribution of L. killicki (syn. L. tropica) and its transmission dynamics. To better understand the evolution of this parasite, its population structure was then compared with that of L. tropica populations from Morocco. In total 198 samples including 85 L. killicki (syn. L. tropica) (from Tunisia, Algeria and Libya) and 113 L. tropica specimens (all from Morocco) were tested. Theses samples were composed of 168 Leishmania strains isolated from human skin lesions, 27 DNA samples from human skin lesion biopsies, two DNA samples from Ctenodactylus gundi bone marrow and one DNA sample from a Phlebotomus sergenti female. The sample was analyzed by using MultiLocus Enzyme Electrophoresis (MLEE) and MultiLocus Microsatellite Typing (MLMT) approaches. Analysis of the MLMT data support the hypothesis that L. killicki (syn. L. tropica) belongs to the L. tropica complex, despite its strong genetic differentiation, and that it emerged from this taxon by a founder effect. Moreover, it revealed a strong structuring in L. killicki (syn. L. tropica) between Tunisia and Algeria and within the different Tunisian regions, suggesting low dispersion of L. killicki (syn. L. tropica) in space and time. Comparison of the L. tropica (exclusively from Morocco) and L. killicki (syn. L. tropica) population structures revealed distinct genetic organizations, reflecting different epidemiological cycles.
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http://dx.doi.org/10.1371/journal.pntd.0004204DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4672892PMC
December 2015

Evolutionary history of Leishmania killicki (synonymous Leishmania tropica) and taxonomic implications.

Parasit Vectors 2015 Apr 1;8:198. Epub 2015 Apr 1.

Département de Parasitologie-Mycologie, Centre National de Référence des Leishmanioses, CHRU de Montpellier, Université de Montpellier, France, 39 avenue Charles FLAHAULT, 34295, Montpellier Cedex 5, France.

Background: The taxonomic status of Leishmania (L.) killicki, a parasite that causes chronic cutaneous leishmaniasis, is not well defined yet. Indeed, some researchers suggested that this taxon could be included in the L. tropica complex, whereas others considered it as a distinct phylogenetic complex. To try to solve this taxonomic issue we carried out a detailed study on the evolutionary history of L. killicki relative to L. tropica.

Methods: Thirty-five L. killicki and 25 L. tropica strains isolated from humans and originating from several countries were characterized using the MultiLocus Enzyme Electrophoresis (MLEE) and the MultiLocus Sequence Typing (MLST) approaches.

Results: The results of the genetic and phylogenetic analyses strongly support the hypothesis that L. killicki belongs to the L. tropica complex. Our data suggest that L. killicki emerged from a single founder event and that it evolved independently from L. tropica. However, they do not validate the hypothesis that L. killicki is a distinct complex. Therefore, we suggest naming this taxon L. killicki (synonymous L. tropica) until further epidemiological and phylogenetic studies justify the L. killicki denomination.

Conclusions: This study provides taxonomic and phylogenetic information on L. killicki and improves our knowledge on the evolutionary history of this taxon.
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http://dx.doi.org/10.1186/s13071-015-0821-6DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4387592PMC
April 2015

Remote sensing, land cover changes, and vector-borne diseases: use of high spatial resolution satellite imagery to map the risk of occurrence of cutaneous leishmaniasis in Ghardaïa, Algeria.

Infect Genet Evol 2014 Dec 7;28:725-34. Epub 2014 Oct 7.

Service d'Eco-Epidémiologie Parasitaire, Institut Pasteur Alger, Algiers, Algeria. Electronic address:

Ghardaïa, central Algeria, experienced a major outbreak of cutaneous leishmaniasis (CL) in 2005. Two Leishmania species occur in this region: Leishmania major (MON-25) and Leishmania killicki (MON-301). The two species are transmitted respectively by the sandflies Phlebotomus papatasi and Phlebotomus sergenti and probably involve rodent reservoirs with different ecologies, suggesting distinct epidemiological patterns and distribution areas. The aims of this study were to establish risk maps for each Leishmania species in Ghardaïa, taking into account the specificities of their vectors and reservoirs biotopes, using land cover and topographical characteristics derived from remote sensing imagery. Using expert and bibliographic knowledge, habitats of vectors and reservoirs were mapped. Hazard maps, defined as areas of presence of both vectors and reservoirs, were then combined with vulnerability maps, defined as areas with human presence, to map the risk of CL occurrence due to each species. The vector habitat maps and risk maps were validated using available entomological data and epidemiological data. The results showed that remote sensing analysis can be used to map and differentiate risk areas for the two species causing CL and identify palm groves and areas bordering the river crossing the city as areas at risk of CL due to L. major, whereas more limited rocky hills on the outskirts of the city are identified as areas at risk of CL due to L. killicki. In the current context of urban development in Ghardaïa, this study provides useful information for the local authorities on the respective risk areas for CL caused by both parasites, in order to take prevention and control measures to prevent future CL outbreaks.
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http://dx.doi.org/10.1016/j.meegid.2014.09.036DOI Listing
December 2014

Heterogeneity of molecular resistance patterns in antimony-resistant field isolates of Leishmania species from the western Mediterranean area.

Antimicrob Agents Chemother 2014 Aug 9;58(8):4866-74. Epub 2014 Jun 9.

Laboratoire de Parasitologie-Mycologie, CHU Timone, UMR MD3 Aix-Marseille Université, Marseille, France.

Antimonials remain the first-line treatment for the various manifestations of leishmaniasis in most areas where the disease is endemic, and increasing cases of therapeutic failure associated with parasite resistance have been reported. In this study, we assessed the molecular status of 47 clinical isolates of Leishmania causing visceral and cutaneous leishmaniasis from Algeria, Tunisia, and southern France. In total, we examined 14 genes that have been shown to exhibit significant variations in DNA amplification, mRNA levels, or protein expression with respect to resistance to antimonials. The gene status of each clinical isolate was assessed via qPCR and qRT-PCR. We then compared the molecular pattern against the phenotype determined via an in vitro sensitivity test of the clinical isolates against meglumine antimoniate, which is considered the reference technique. Our results demonstrate significant DNA amplification and/or RNA overexpression in 56% of the clinical isolates with the resistant phenotype. All clinical isolates that exhibited significant overexpression of at least 2 genes displayed a resistant phenotype. Among the 14 genes investigated, 10 genes displayed either significant amplification or overexpression in at least 1 clinical isolate; these genes are involved in several metabolic pathways. Moreover, various gene associations were observed depending on the clinical isolates, supporting the multifactorial nature of Leishmania resistance. Molecular resistance features were found in the 3 Leishmania species investigated (Leishmania infantum, Leishmania major, and Leishmania killicki). To our knowledge, this is the first report of the involvement of molecular resistance genes in field isolates of Leishmania major and Leishmania killicki with the resistance phenotype.
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http://dx.doi.org/10.1128/AAC.02521-13DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4136049PMC
August 2014

[A social program for the control of zoonotic cutaneous leishmaniasis in M'Sila, Algeria].

Sante Publique 2012 Nov-Dec;24(6):511-22

Departement de Biologie, Universite de M'Sila, Algerie.

Zoonotic cutaneous leishmaniasis due to Leishmania major is a serious public health problem in Algeria. On average, 10,000 new cases are reported every year among the 15 million people at risk of infection. With an annual incidence of 561.8 per 100,000 inhabitants, M'Sila has seen the worst outbreak of the disease in Algeria since the historic outbreak in Biskra. The main reservoir of the disease is Psammomys obesus, a gerbil that feeds exclusively on Chenopodiaceae, a salt-tolerant plant under which it makes its burrow. Removing these plants around houses within a radius of 300 meters is one of the most effective control measures. As part of a social program of public works, a pilot project aimed at controlling the disease was undertaken in 2003 in the five worst affected cities in M'Sila. 396 unemployed young people were recruited to remove the plants before the transmission season. Over 3,600 hectares were treated. The number of cases decreased from 1,391 in 2003 to 965 in 2004 (31% reduction). These measures need to be implemented in all endemic areas of the country to better assess their effectiveness in preventing the disease.
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April 2013

Culex pipiens, an experimental efficient vector of West Nile and Rift Valley fever viruses in the Maghreb region.

PLoS One 2012 31;7(5):e36757. Epub 2012 May 31.

Institut Pasteur du Maroc, Laboratoire des Maladies Vectorielles, Casablanca, Maroc.

West Nile fever (WNF) and Rift Valley fever (RVF) are emerging diseases causing epidemics outside their natural range of distribution. West Nile virus (WNV) circulates widely and harmlessly in the old world among birds as amplifying hosts, and horses and humans as accidental dead-end hosts. Rift Valley fever virus (RVFV) re-emerges periodically in Africa causing massive outbreaks. In the Maghreb, eco-climatic and entomologic conditions are favourable for WNV and RVFV emergence. Both viruses are transmitted by mosquitoes belonging to the Culex pipiens complex. We evaluated the ability of different populations of Cx. pipiens from North Africa to transmit WNV and the avirulent RVFV Clone 13 strain. Mosquitoes collected in Algeria, Morocco, and Tunisia during the summer 2010 were experimentally infected with WNV and RVFV Clone 13 strain at titers of 10(7.8) and 10(8.5) plaque forming units/mL, respectively. Disseminated infection and transmission rates were estimated 14-21 days following the exposure to the infectious blood-meal. We show that 14 days after exposure to WNV, all mosquito st developed a high disseminated infection and were able to excrete infectious saliva. However, only 69.2% of mosquito strains developed a disseminated infection with RVFV Clone 13 strain, and among them, 77.8% were able to deliver virus through saliva. Thus, Cx. pipiens from the Maghreb are efficient experimental vectors to transmit WNV and to a lesser extent, RVFV Clone 13 strain. The epidemiologic importance of our findings should be considered in the light of other parameters related to mosquito ecology and biology.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0036757PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3365064PMC
October 2012

DNA barcodes confirm the presence of a single member of the Anopheles maculipennis group in Morocco and Algeria: An. sicaulti is conspecific with An. labranchiae.

Acta Trop 2011 Apr 21;118(1):6-13. Epub 2010 Dec 21.

Institut National d'Hygiène, Agdal, Rabat, Morocco.

Anopheles labranchiae Falleroni is the only member of the Maculipennis Group known to occur in northern Africa; however, confusion exists as to the taxonomic status of its junior synonym, An. sicaulti Roubaud (type locality: near Rabat, Morocco). Based on morphological and behavioural distinctions, it has been suggested that Moroccan populations have been isolated from other North African populations by the Atlas Mountains, and that Moroccan populations may represent An. sicaulti, originally described as a variety of An. maculipennis Meigen. DNA barcodes (658bp of the mitochondrial COI gene) obtained from 89 An. maculipennis s.l. collected in Morocco (n=79) and Algeria (n=10) in 2007 and 2008 were used to determine if Moroccan populations are genetically isolated from those east of the Atlas Mountains (Algeria), and whether there is molecular evidence to support the presence of more than one member of the Maculipennis Group in the region. No evidence for speciation was found between Moroccan and Algerian populations, or within populations in northern Morocco. Moreover shared COI haplotypes between Algeria and Morocco indicate ongoing gene flow between populations in these countries, suggesting that the Atlas Mountains are not a boundary to gene flow in An. labranchiae. The synonymy of An. sicaulti with An. labranchiae is confirmed. That An. labranchiae comprises the same species in these North African countries is important for malaria control.
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http://dx.doi.org/10.1016/j.actatropica.2010.12.006DOI Listing
April 2011

Plasmodium falciparum malaria, southern Algeria, 2007.

Emerg Infect Dis 2010 Feb;16(2):301-3

Institut Pasteur, Algiers, Algeria.

An outbreak of Plasmodium falciparum malaria occurred in Tinzaouatine in southern Algeria in 2007. The likely vector, Anopheles gambiae mosquitoes, had not been detected in Algeria. Genes for resistance to chloroquine were detected in the parasite. The outbreak shows the potential for an increase in malaria vectors in Algeria.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2958010PMC
http://dx.doi.org/10.3201/eid1602.090914DOI Listing
February 2010

Description of a dermatropic Leishmania close to L. killicki (Rioux, Lanotte & Pratlong 1986) in Algeria.

Trans R Soc Trop Med Hyg 2009 Jul 22;103(7):716-20. Epub 2009 May 22.

Service d'Eco-Epidémiologie Parasitaire, Centre National de Référence des Leishmania, Institut Pasteur d'Algérie, Annexe de Sidi Fredj, Dely Ibrahim, 16000 Alger, Algeria.

Cutaneous leishmaniasis (CL) is endemic in Algeria where two forms have been previously described, the sporadic form caused by Leishmania infantum in the north and the cutaneous form caused by L. major in central and southern parts of the country. During 2005, a CL outbreak occurred in the province of Ghardaïa, located in the north of Sahara, where 2040 cases were recorded, of which several were from urban areas. Six strains isolated from patients with active lesions were identified by isoenzyme electrophoresis and by molecular typing using systematic sequencing of a large subunit of the RNA polymerase. Four of the strains belonged to a new zymodeme, MON-301, close to L. killicki MON-8. The two other isolates were identified as L. major zymodeme MON-25. The new dermatropic Leishmania close to L. killicki is reported for the first time in Algeria and coexists sympatrically with L. major MON-25 in the region of Ghardaïa where they occur in their usual vectors of Phlebotomus papatasi (L. major) and P. sergenti (L. tropica). This new parasite demonstrates the need for further investigations to elucidate the life cycle and transmission of the emergent disease and to evaluate its phylogenetic position in the taxonomy of Leishmania.
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http://dx.doi.org/10.1016/j.trstmh.2009.04.013DOI Listing
July 2009
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