FREE SHIPPING over $40
OVER 10000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Veterinary research communications2025; 49(6); 328; doi: 10.1007/s11259-025-10904-8

Serological, molecular, and epidemiological assessment of Leishmania spp. in equids within a hotspot endemic area for cutaneous leishmaniasis in southeastern Algeria.

Abstract: Cutaneous leishmaniosis remains a major public health concern in Algeria, predominantly affecting humans and dogs. However, the role of other animal hosts, particularly equids, in the transmission cycle remains underexplored. This cross-sectional study presents the first systematic investigation of Leishmania spp. exposure and infection in equids, along with associated environmental and host factors, from southeastern Algeria, El Oued Wilaya, the third most affected region for human cutaneous leishmaniasis in the country. A total of 122 equids from eight communes were clinically examined and tested using three diagnostic tools: indirect fluorescent antibody test, conventional PCR targeting the ITS1 region, and real-time qPCR targeting the kDNA minicircle. Despite the absence of clinical symptoms, serological analysis revealed a 22.13% seropositivity rate for anti-Leishmania infantum IgG. ITS1-PCR and kDNA qPCR detected parasite DNA in 13.93% and 7.38% of samples, respectively. PCR-RFLP analysis confirmed the presence of Leishmania (Leishmania) infantum. Phylogenetic analysis revealed intraspecific divergence among isolates, with two sequences clustering in a distinct clade closely related to a Brazilian strain, indicating unexpected genetic diversity among Algerian isolates. Key associated factors included proximity to human settlements and age-related susceptibility. The detection of subclinical infections in equids suggests a potential but underrecognized epidemiological role in endemic areas. Although their reservoir competence remains unconfirmed, this evidence warrants further research on vector competence, molecular characterization, and entomological surveillance. Integrating equids into One Health surveillance frameworks could help clarify their role and guide targeted control strategies in Algeria where transmission dynamics of leishmaniosis may vary.
© 2025. The Author(s), under exclusive licence to Springer Nature B.V.
Get Full Text
Publication Date: 2025-09-25 PubMed ID: 40996657PubMed Central: 7064173DOI: 10.1007/s11259-025-10904-8Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
LinkedInCopy
  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

Overview

  • This study investigates the exposure to and infection by Leishmania parasites in horses and donkeys (equids) in southeastern Algeria, a region highly affected by cutaneous leishmaniasis.
  • The research aims to understand the role of equids in the disease’s transmission cycle by combining serological and molecular diagnostics with epidemiological analysis.

Research Context and Objectives

  • Cutaneous leishmaniasis is a significant public health problem in Algeria, primarily known to affect humans and dogs.
  • There is limited knowledge about how other animals, especially equids (horses and donkeys), may contribute to the disease’s maintenance and spread.
  • The study focuses on El Oued Wilaya in southeastern Algeria, the third most affected area for human cutaneous leishmaniasis in the country.
  • Its primary objective is to assess the presence of Leishmania parasites in equids and identify environmental and host factors associated with infection.

Study Design and Methods

  • Sampling: A total of 122 equids were sampled from 8 different communes within the hotspot region.
  • Clinical examination: Animals were examined for symptoms of leishmaniasis, though none showed clinical signs.
  • Diagnostic tools used:
    • Indirect fluorescent antibody test (IFA) to detect antibodies against Leishmania infantum.
    • Conventional PCR targeting the internal transcribed spacer 1 (ITS1) region of the parasite DNA.
    • Real-time quantitative PCR (qPCR) targeting the kDNA minicircle, a highly repetitive genetic element for sensitive detection.

Key Findings

  • Serological results: 22.13% of equids were positive for anti-Leishmania infantum IgG antibodies, indicating exposure to the parasite.
  • Molecular detection:
    • ITS1-PCR detected parasite DNA in 13.93% of samples.
    • kDNA qPCR detected parasite DNA in 7.38% of samples, confirming active or recent infections.
  • Parasite species identification through PCR-RFLP confirmed the presence of Leishmania (Leishmania) infantum.
  • Genetic diversity: Phylogenetic analysis showed intraspecific diversity among isolates, with some clustering into a distinct clade closer to a Brazilian strain, which suggests unexpected genetic variation among Algerian parasites.

Associated Epidemiological Factors

  • Proximity to human settlements: Equids located near human populations had higher chances of exposure or infection, pointing to shared transmission environments.
  • Age susceptibility: Age-related differences in infection rates indicated variability in vulnerability or exposure over the equids’ lifespans.

Implications and Recommendations

  • The detection of subclinical (asymptomatic) infections in equids suggests these animals might play a previously underrecognized role in maintaining or spreading Leishmania parasites in endemic areas.
  • Although it is not yet confirmed whether equids can act as reservoirs (i.e., sustain and transmit the infection), their involvement in transmission cycles merits further investigation.
  • Future research should focus on:
    • Vector competence studies to see if sandflies transmit Leishmania from equids.
    • Further molecular characterization of circulating parasite strains.
    • Entomological surveillance to monitor sandfly populations and infection rates.
  • Integrating equids into One Health frameworks—considering human, animal, and environmental health together—may improve surveillance and guide targeted control strategies in Algeria.
  • This approach is especially important given the possible variations in leishmaniasis transmission dynamics across different regions and host species.

Cite This Article

APA
Djellouli M, Eddaikra N, Beneldjouzi A, Benikhlef R, Ghimire R, Wilkins M, Lafri I. (2025). Serological, molecular, and epidemiological assessment of Leishmania spp. in equids within a hotspot endemic area for cutaneous leishmaniasis in southeastern Algeria. Vet Res Commun, 49(6), 328. https://doi.org/10.1007/s11259-025-10904-8

Publication

Veterinary research communications
ISSN: 1573-7446
NlmUniqueID: 8100520
Country: Switzerland
Language: English
Volume: 49
Issue: 6
Pages: 328

Researcher Affiliations

Djellouli, Meriem
  • Research Laboratory of Exploration and Valorization of Steppe Ecosystems, Ziane Achour Djelfa University, Djelfa, 17000, Algeria.
  • National Institute of Veterinary Medicine, Algiers, 16000, Algeria.
Eddaikra, Naouel
  • Laboratory of Parasitic Eco-epidemiology and Population Genetics, Pasteur Institute of Algeria, Algiers, 16030, Algeria.
Beneldjouzi, Assia
  • Laboratory of Parasitic Eco-epidemiology and Population Genetics, Pasteur Institute of Algeria, Algiers, 16030, Algeria.
Benikhlef, Razika
  • Laboratory of Parasitic Eco-epidemiology and Population Genetics, Pasteur Institute of Algeria, Algiers, 16030, Algeria.
Ghimire, Ramjee
  • Institute for Global Health, Michigan State University, East Lansing, Michigan, 48824, USA.
Wilkins, Melinda
  • Department of Veterinary Population Medicine, College of Veterinary Medicine, University of Minnesota, Saint Paul, Minnesota, 55102, USA.
Lafri, Ismail
  • Department of Microbiology and Veterinary Pathology, Laboratory of Veterinary Bacteriology, Pasteur Institute of Algeria, Algiers, 16000, Algeria. lafrismail@gmail.com.

MeSH Terms

  • Animals
  • Algeria / epidemiology
  • Horses
  • Leishmaniasis, Cutaneous / veterinary
  • Leishmaniasis, Cutaneous / epidemiology
  • Leishmaniasis, Cutaneous / parasitology
  • Horse Diseases / epidemiology
  • Horse Diseases / parasitology
  • Cross-Sectional Studies
  • Female
  • Phylogeny
  • Male
  • Leishmania infantum / isolation & purification
  • Leishmania infantum / genetics
  • Leishmania / isolation & purification
  • Leishmania / genetics

Conflict of Interest Statement

Declarations. Consent for publication: All authors gave their consent for publication. Competing interests: The authors declare no competing interests. Ethics approval: The Ethics Committee of the Faculty of Nature and Life Sciences at Ziane Achour University in Algeria, approved the study, which was conducted in compliance with national guidelines (Process number: DOC1721SNV19055/EVSE2022). A prior informed verbal consent of the animal owners was obtained to participate to this study before sample collection.

References

This article includes 55 references
  1. Abbate MJ, Maia C, Pereira A. Identification of trypanosomatids and blood feeding preferences of phlebotomine sand fly species common in Sicily, Southern Italy. PLoS ONE 15(3):e0229536.
    doi: 10.1371/JOURNAL.PONE.0229536pubmed: 32155171pmc: 7064173google scholar: lookup
  2. Abidi H. Application of the GIS to determine the physico-chemical quality of the waters of the boreholes intended for the AEP in the Suf region. Dissertation, University of Algeria.
  3. Aguilar CM, Rangel EF, Garcia L. Zoonotic cutaneous leishmaniasis due to Leishmania (Viannia) Braziliensis associated with domestic animals in Venezuela and Brazil. Mem Inst Oswaldo Cruz 84(1):19–28.
    doi: 10.1590/S0074-02761989000100005pubmed: 2319948google scholar: lookup
  4. Akhoundi M, Kuhls K, Cannet A. A historical overview of the classification, evolution, and dispersion of Leishmania parasites and sandflies. PLoS Negl Trop Dis .
    doi: 10.1371/journal.pntd.0004349pubmed: 26937644pmc: 4777430google scholar: lookup
  5. Alcover MM, Ribas A, Guillén MC. Wild mammals as potential silent reservoirs of Leishmania infantum in a mediterranean area. Prev Vet Med .
    doi: 10.1016/j.prevetmed.2019.104874pubmed: 31901603google scholar: lookup
  6. Alten B, Maia C, Afonso MO. Seasonal dynamics of phlebotomine sand fly species proven vectors of mediterranean leishmaniasis caused by Leishmania infantum. PLoS Negl Trop Dis 10(2).
    doi: 10.1371/journal.pntd.0004458google scholar: lookup
  7. . Monography Wilaya of El Oued. .
  8. Benassi JC, Benvenga GU, Ferreira HL. Molecular and serological detection of Leishmania spp. in horses from an endemic area for canine visceral leishmaniasis in southeastern Brazil. Pesq Vet Bras 38(6):1058–63.
    doi: 10.1590/1678-5150-PVB-5214google scholar: lookup
  9. . IndiMag Pathogen Kit Handbook for automated purification of viral RNA and DNA and bacterial DNA from animal samples using IndiMag 48, KingFisherTM Flex, BioSprint 96 or equivalent workstation. pp 25–30.
  10. Biral NV, Santos HA, Senne NA. A Cross-sectional study of Leishmania spp. In draft horses from the Distrito federal, brazil: seroprevalence, Spatial distribution, and associated factors. Prev Vet Med 195(October):105467.
    doi: 10.1016/j.prevetmed.2021.105467pubmed: 34416652google scholar: lookup
  11. Calzolari M, Romeo G, Bergamini F. Host preference and Leishmania infantum natural infection of the sand fly Phlebotomus perfiliewi in northern Italy. Acta Trop 226:106246.
    doi: 10.1016/j.actatropica.2021.106246pubmed: 34843690google scholar: lookup
  12. Ceccarelli M, Buffi G, Diotallevi A. Evaluation of a KDNA-based qPCR assay for the detection and quantification of old world Leishmania species. Microorganisms 8(12):2006.
    doi: 10.3390/MICROORGANISMS8122006pubmed: 33339158pmc: 7765608google scholar: lookup
  13. Chargui N, Slama D, Haouas N, Rmadi L, Babba H. Transmission cycle analysis in a Leishmania infantum focus: infection rates and blood meal origins in sand flies (Diptera: Psychodidae). J Vector Ecol 43(2):321–327.
    doi: 10.1111/jvec.12316pubmed: 30408299google scholar: lookup
  14. Cosma C, Maia C, Khan N, Infantino M, Del Riccio M. Leishmaniasis in humans and animals: a one health approach for surveillance, prevention and control in a changing world. Trop Med Infect Dis .
    doi: 10.3390/TROPICALMED9110258pubmed: 39591264pmc: 11598728google scholar: lookup
  15. dos Marques S, Gomes LH, da Rocha LI. Low Parasite Load Estimated by QPCR in a Cohort of Children Living in Urban Area Endemic for Visceral Leishmaniasis in Brazil. PLOS Negl Trop Dis 6(12):e1955.
    doi: 10.1371/JOURNAL.PNTD.0001955google scholar: lookup
  16. Drissi G, Mhadhbi M, Sassi L. Epidemiological survey of vector-borne infections in equids from northern Tunisia. Rev Sci Tech Off Int Epiz 37(3):1021–1027.
    doi: 10.20506/rst.37.3.2904google scholar: lookup
  17. DSP El Oued (2023) Directorate of health and population of El Oued Wilaya. Ministry of Health, Population and Hospital Reform, Algeria
  18. Dubie T, Mohammed Y. Review on the role of host immune response in protection and immunopathogenesis during cutaneous leishmaniasis infection. J Immunol Res 2020:1–12.
    doi: 10.1155/2020/2496713google scholar: lookup
  19. Escobar TA, Döwich G, Cantele LC. Applications of polymerase chain reaction for the detection of equine Leishmania sp. infection. Semina Cienc Agrar 41(1):199.
    doi: 10.5433/1679-0359.2020v41n1p199google scholar: lookup
  20. Fernández-Bellon H, Solano-Gallego L, Bardagí M. Immune response to Leishmania infantum in healthy horses in Spain. Vet Parasitol 135(2):181–185.
    doi: 10.1016/j.vetpar.2005.09.007pubmed: 16213661google scholar: lookup
  21. Gabriel A, Valério-Bolas A, Palma-Marques J. Cutaneous Leishmaniasis: The Complexity of Host’s Effective Immune Response against a Polymorphic Parasitic Disease. J Immunol Res 2019:2603730.
    doi: 10.1155/2019/2603730google scholar: lookup
  22. Gallo-Francisco PH, Brocchi M, Giorgio S. Leishmania and its relationships with bacteria. Future Microbiol 17(3):199–218.
    doi: 10.2217/FMB-2021-0133pubmed: 35040703google scholar: lookup
  23. Gazzonis AL, Morganti G, Porcellato I. Detection of Leishmania spp. in chronic dermatitis: retrospective study in exposed horse populations. Pathogens 11(6):634.
    doi: 10.3390/pathogens11060634pubmed: 35745488pmc: 9227255google scholar: lookup
  24. Guillen MC, Alcover M, Borruel M N. Leishmania infantum asymptomatic infection in inflammatory bowel disease patients under anti-TNF therapy. .
    doi: 10.1016/j.heliyon.2020.e03940google scholar: lookup
  25. Infoclimat (2024) Climatologie de l’année 2023 à El Oued – Infoclimat. https://www.infoclimat.fr/climatologie/annee/2023/el-oued/valeurs/60559.html . Accessed: 18 April 2025
  26. INSP (2024) Monthly epidemiological report publications - National Institute of Public Health (INSP). https://www.insp.dz/index.php/Non-categorise/rem.html . Accessed: 20 January 2024
  27. Kaba Z. Trends of cutaneous leishmaniasis in Western Ethiopia: a retrospective study. Biosci Biotechnol Res Commun 17(3):154–162.
    doi: 10.21786/BBRC/17.3.5google scholar: lookup
  28. Kenubih A, Dagnachew S, Almaw G. Preliminary survey of domestic animal visceral leishmaniasis and risk factors in north-west Ethiopia. Trop Med Int Health 20(2):205–210.
    doi: 10.1111/tmi.12418pubmed: 25327874google scholar: lookup
  29. Khelifi Touhami NA, Ouchene N, Ouchetati I, Naghib I. Animal leishmaniasis in Algeria: a systematic review and meta-analysis. Comp Immunol Microbiol Infect Dis 93:101930.
    doi: 10.1016/j.cimid.2022.101930pubmed: 36584413google scholar: lookup
  30. Khezzani B, Bouchemal S. Demographic and spatio-temporal distribution of cutaneous leishmaniasis in the Souf Oasis (Eastern South of Algeria): results of 13 years. Acta Trop 166:74–80.
    doi: 10.1016/j.actatropica.2016.11.012pubmed: 27840067google scholar: lookup
  31. Kimutai A, Ngure PK, Tonui WK, Gicheru MM, Nyamwamu LB. Leishmaniasis in northern and western Africa: a review. Afr J Infect Dis 3(1):14–25.
    doi: 10.4314/AJID.V3I1.55077google scholar: lookup
  32. Kuhls K, Alam MZ, Cupolillo E. Comparative microsatellite typing of new world Leishmania infantum reveals low heterogeneity among populations and its recent old-world origin. PLoS Negl Trop Dis .
    doi: 10.1371/JOURNAL.PNTD.0001155pubmed: 21666787pmc: 3110170google scholar: lookup
  33. Limeira CH, Alves CJ, de Azevedo SS, de Melo MA. Clinical aspects and diagnosis of leishmaniasis in equids: a systematic review and meta-analysis. Rev Bras Parasitol Vet 28(4):574–81.
    doi: 10.1590/s1984-29612019074pubmed: 31596317google scholar: lookup
  34. MADR (2022) Law No. 88 – 08 of January 26, 1988 - Relating to the activities of veterinary medicine and the protection of animal health. https://fr.madr.gov.dz/lois-et-ordonnances . Accessed 01 January 2022
  35. Mahachi KG, Ozanne M, Bourdeau P. Comparison of ELISA and IFAT for Leishmania infantum by European and middle Eastern diagnostic laboratories. Parasit Vectors .
    doi: 10.1186/S13071-024-06631-9pubmed: 39734221pmc: 11684067google scholar: lookup
  36. Mary C, Faraut F, Lascombe L, Dumon H. Quantification of Leishmania infantum DNA by a real-time PCR assay with high sensitivity. J Clin Microbiol 42(11):5249–5255.
    doi: 10.1128/JCM.42.11.5249-5255.2004pubmed: 15528722pmc: 525214google scholar: lookup
  37. Medkour H, Laidoudi Y, Lafri I, Bitam I, Mediannikov O, Davoust B. Canine leishmaniosis and first report of Leishmania infantum in the blood of equids in Kabylia (Algeria). Int J Infect Dis 79(S1):1–150.
    doi: 10.1016/j.ijid.2018.11.287google scholar: lookup
  38. Merdekios B, Pareyn M, Tadesse D, Eligo N, Kassa M, JBKM, Leirs H. Evaluation of conventional and four real-time PCR methods for the detection of Leishmania on field-collected samples in Ethiopia. PLoS Negl Trop Dis 15(1):e0008903.
    doi: 10.1371/JOURNAL.PNTD.0008903pubmed: 33434190pmc: 7802924google scholar: lookup
  39. Merrouchi L, Boualem B, Benziouche SE. The agricultural system in the Oued-Righ valley (Southeast Algeria): characteristics and functioning. Qadisiyah J Agric Sci 13(1):59–70.
    doi: 10.33794/QJAS.2022.134892.1070google scholar: lookup
  40. Mhadhbi M, Sassi A. Infection of the equine population by Leishmania parasites. Equine Vet J 52(1):28–33.
    doi: 10.1111/evj.13178pubmed: 31498914google scholar: lookup
  41. Muñoz C, Risueño J, Pérez-Cutillas P, Bernal LJ, Ortiz JM, Ruiz de Ybáñez R, Sánchez-López PF. Density assessment and reporting for Phlebotomus perniciosus and other sand fly species in Periurban residential estates in Spain. Parasitol Res 120(9):3091–3103.
    doi: 10.1007/S00436-021-07270-0/TABLES/5pubmed: 34405280pmc: 8397643google scholar: lookup
  42. Open Database License 1.0 (2024) Download free shapefiles layers of Algeria | GIS English. https://gisenglish.geojamal.com/2018/05/download-free-shapefiles-layers-of_81.html Assessed 28 July 2024
  43. Pala S, Martínez-Sáez L, Llobat L, Marín-García PJ. Prevalence and factors associated with Leishmania spp. and Toxoplasma gndii infections in apparently healthy horses in Eastern Spain. Res Vet Sci .
    doi: 10.1016/j.rvsc.2024.105236pubmed: 38531238google scholar: lookup
  44. Sarkari B, Ashrafmansouri M, Reza Hatam G. Performance of an ELISA and indirect Immunofluorescence assay in serological diagnosis of zoonotic cutaneous leishmaniasis in Iran. Interdiscip Perspect Infect Dis .
    doi: 10.1155/2014/505134google scholar: lookup
  45. Schönian G, Nasereddin A, Dinse N, Schweynoch C, Schallig Henk DFH, Presber W, Jaffe CL. PCR diagnosis and characterization of Leishmania in local and imported clinical samples. Diagn Microbiol Infect Dis 47(1):349–358.
    doi: 10.1016/S0732-8893(03)00093-2pubmed: 12967749google scholar: lookup
  46. Schönian G, Mauricio I, Gramiccia M, Cañavate C, Boelaert M, Dujardin JD. Leishmaniases in the Mediterranean in the era of molecular epidemiology. Trends Parasitol 24(3):135–142.
    doi: 10.1016/J.PT.2007.12.006pubmed: 18262469google scholar: lookup
  47. Sidstedt M, Hedman J, Romsos EL, Waitara L. Inhibition mechanisms of hemoglobin, Immunoglobulin G, and whole blood in digital and real-time PCR. Anal Bioanal Chem 410(10):2569.
    doi: 10.1007/S00216-018-0931-Zpubmed: 29504082pmc: 5857286google scholar: lookup
  48. Tamura K, Peterson D, Peterson N. MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol 28(10):2731–2739.
    doi: 10.1093/MOLBEV/MSR121pubmed: 21546353pmc: 3203626google scholar: lookup
  49. Thakur S, Joshi J, Kaur S. Leishmaniasis diagnosis: an update on the use of parasitological, immunological and molecular methods. J Parasit Dis 44(2):253–272.
    doi: 10.1007/S12639-020-01212-W/TABLES/2pubmed: 32419743pmc: 7223249google scholar: lookup
  50. Truppel JH, Otomura F, Teodoro U, Massafera R, Costa-Ribeiro MCVDa, Catarino CM. Can equids be a reservoir of Leishmania Braziliensis in endemic areas. PLoS ONE 9(4):e93731.
    doi: 10.1371/journal.pone.0093731pubmed: 24721908pmc: 3983081google scholar: lookup
  51. Van der Auwera G, Dujardina JD. Species typing in dermal leishmaniasis. Clin Microbiol Rev 28(2):265.
    doi: 10.1128/CMR.00104-14pubmed: 25672782pmc: 4402951google scholar: lookup
  52. Vedovello Filho D, Jorge FA, Lonardoni MV. American cutaneous leishmaniasis in horses from endemic areas in the north-central mesoregion of Paraná state, Brazil. Zoonoses Public Health 55(3):149–155.
    doi: 10.1111/j.1863-2378.2008.01106.xpubmed: 18331518google scholar: lookup
  53. . Global leishmaniasis surveillance: 2019–2020, a Baseline for the 2030 Roadmap. September 3, 2021.
  54. . Leishmaniasis - WHO Guidelines. January 12, 2023.
  55. . Terrestrial Manual. .

Citations

This article has been cited 0 times.
Subscribe to our newsletter
Join 99,911 horse owners like you who receive our email updates on horse health and nutrition.