Background Phlebotomine sand flies are known to transmit Leishmania parasites, bacteria and viruses that affect humans and animals in many countries worldwide. Precise sand fly identification is essential to prevent phlebotomine-borne diseases. Over the past two decades, progress in matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) has emerged as an accurate tool for arthropod identification. The objective of the present study was to investigate the usefulness of MALDI-TOF MS as a tool for identifying field-caught phlebotomine. Methodology/Principal Findings Sand flies were captured in four sites in north Algeria. A subset was morphologically and genetically identified. Six species were found in these areas and a total of 28 stored frozen specimens were used for the creation of the reference spectrum database. The relevance of this original method for sand fly identification was validated by two successive blind tests including the morphological identification of 80 new specimens which were stored at -80°C, and 292 unknown specimens, including engorged specimens, which were preserved under different conditions. Intra-species reproducibility and inter-species specificity of the protein profiles were obtained, allowing us to distinguish specimens at the gender level. Querying of the sand fly database using the MS spectra from the blind test groups revealed concordant results between morphological and MALDI-TOF MS identification. However, MS identification results were less efficient for specimens which were engorged or stored in alcohol. Identification of 362 phlebotomine sand flies, captured at four Algerian sites, by MALDI-TOF MS, revealed that the subgenus Larroussius was predominant at all the study sites, except for in M’sila where P. (Phlebotomus) papatasi was the only sand fly species detected. Conclusion The present study highlights the application of MALDI-TOF MS for monitoring sand fly fauna captured in the field. The low cost, reliability and rapidity of MALDI-TOF MS analyses opens up new ways in the management of phlebotomine sand fly-borne diseases.
References
[1]
Maroli M, Feliciangeli MD, Bichaud L, Charrel RN, Gradoni L (2013) Phlebotomine sandflies and the spreading of leishmaniases and other diseases of public health concern. Med Vet Entomol 27: 123–147. doi: 10.1111/j.1365-2915.2012.01034.x. pmid:22924419
[2]
Alvar J, Velez ID, Bern C, Herrero M, Desjeux P, Cano J, Jannin J, den BM (2012) Leishmaniasis worldwide and global estimates of its incidence. PLoS One 7: e35671. doi: 10.1371/journal.pone.0035671. PONE-D-11-24894 [pii]. pmid:22693548
[3]
Chaara D, Haouas N, Dedet JP, Babba H, Pratlong F (2014) Leishmaniases in Maghreb: an endemic neglected disease. Acta Trop 132: 80–93. S0001-706X(13)00370-7 [pii]; doi: 10.1016/j.actatropica.2013.12.018. pmid:24412727
[4]
Aoun K, Bouratbine A (2014) Cutaneous leishmaniasis in North Africa: a review. Parasite 21: 14. doi: 10.1051/parasite/2014014. parasite130072 [pii]. pmid:24626301
[5]
Killick-Kendrick R, Tang Y, Killick-Kendrick M, Sang DK, Sirdar MK, Ke L, Ashford RW, Schorscher J, Johnson RH (1991) The identification of female sandflies of the subgenus Larroussius by the morphology of the spermathecal ducts. Parassitologia 33 Suppl: 335–347. pmid:1841225
[6]
Dvorak V, Halada P, Hlavackova K, Dokianakis E, Antoniou M, Volf P (2014) Identification of phlebotomine sand flies (Diptera: Psychodidae) by matrix-assisted laser desorption/ionization time of flight mass spectrometry. Parasit Vectors 7: 21. 1756-3305-7-21 [pii]; doi: 10.1186/1756-3305-7-21. pmid:24423215
[7]
Depaquit J, Ferte H, Leger N, Killick-Kendrick R, Rioux JA, Killick-Kendrick M, Hanafi HA, Gobert S (2000) Molecular systematics of the phlebotomine sandflies of the subgenus Paraphlebotomus (diptera, psychodidae, phlebotomus) based on ITS2 rDNA sequences. Hypotheses Of dispersion and speciation. Insect Mol Biol 9: 293–300. imb179 [pii]. pmid:10886413 doi: 10.1046/j.1365-2583.2000.00179.x
[8]
Depaquit J, Lienard E, Verzeaux-Griffon A, Ferte H, Bounamous A, Gantier JC, Hanafi HA, Jacobson RL, Maroli M, Moin-Vaziri V, Muller F, Ozbel Y, Svobodova M, Volf P, Leger N (2008) Molecular homogeneity in diverse geographical populations of Phlebotomus papatasi (Diptera, Psychodidae) inferred from ND4 mtDNA and ITS2 rDNA Epidemiological consequences. Infect Genet Evol 8: 159–170. S1567-1348(07)00196-7 [pii]; doi: 10.1016/j.meegid.2007.12.001. pmid:18243814
[9]
Terayama Y, Kato H, Gomez EA, Uezato H, Calvopina M, Iwata H, Hashiguchi Y (2008) Molecular typing of sand fly species (Diptera, Psychodidae, Phlebotominae) from areas endemic for Leishmaniasis in Ecuador by PCR-RFLP of 18S ribosomal RNA gene. J Vet Med Sci 70: 907–913. JST.JSTAGE/jvms/70.907 [pii]. pmid:18840964 doi: 10.1292/jvms.70.907
[10]
Contreras Gutierrez MA, Vivero RJ, Velez ID, Porter CH, Uribe S (2014) DNA barcoding for the identification of sand fly species (Diptera, Psychodidae, Phlebotominae) in Colombia. PLoS One 9: e85496. doi: 10.1371/journal.pone.0085496. PONE-D-13-30108 [pii]. pmid:24454877
[11]
Mellmann A, Bimet F, Bizet C, Borovskaya AD, Drake RR, Eigner U, Fahr AM, He Y, Ilina EN, Kostrzewa M, Maier T, Mancinelli L, Moussaoui W, Prevost G, Putignani L, Seachord CL, Tang YW, Harmsen D (2009) High interlaboratory reproducibility of matrix-assisted laser desorption ionization-time of flight mass spectrometry-based species identification of nonfermenting bacteria. J Clin Microbiol 47: 3732–3734. JCM.00921-09 [pii]; doi: 10.1128/JCM.00921-09. pmid:19776231
[12]
Santos C, Paterson RR, Venancio A, Lima N (2010) Filamentous fungal characterizations by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. J Appl Microbiol 108: 375–385. JAM4448 [pii]; doi: 10.1111/j.1365-2672.2009.04448.x. pmid:19659699
[13]
Stevenson LG, Drake SK, Murray PR (2010) Rapid identification of bacteria in positive blood culture broths by matrix-assisted laser desorption ionization-time of flight mass spectrometry. J Clin Microbiol 48: 444–447. JCM.01541-09 [pii]; doi: 10.1128/JCM.01541-09. pmid:19955282
[14]
Perera MR, Vanstone VA, Jones MG (2005) A novel approach to identify plant parasitic nematodes using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Rapid Commun Mass Spectrom 19: 1454–1460. doi: 10.1002/rcm.1943. pmid:15880621
[15]
Karger A, Kampen H, Bettin B, Dautel H, Ziller M, Hoffmann B, Suss J, Klaus C (2012) Species determination and characterization of developmental stages of ticks by whole-animal matrix-assisted laser desorption/ionization mass spectrometry. Ticks Tick Borne Dis 3: 78–89. S1877-959X(11)00089-6 [pii]; doi: 10.1016/j.ttbdis.2011.11.002. pmid:22487425
[16]
Yssouf A, Flaudrops C, Drali R, Kernif T, Socolovschi C, Berenger JM, Raoult D, Parola P (2013) Matrix-assisted laser desorption ionization-time of flight mass spectrometry for rapid identification of tick vectors. J Clin Microbiol 51: 522–528. JCM.02665-12 [pii]; doi: 10.1128/JCM.02665-12. pmid:23224087
[17]
Muller P, Pfluger V, Wittwer M, Ziegler D, Chandre F, Simard F, Lengeler C (2013) Identification of cryptic Anopheles mosquito species by molecular protein profiling. PLoS One 8: e57486. doi: 10.1371/journal.pone.0057486. PONE-D-12-33530 [pii]. pmid:23469000
[18]
Dieme C, Yssouf A, Vega-Rua A, Berenger JM, Failloux AB, Raoult D, Parola P, Almeras L (2014) Accurate identification of Culicidae at aquatic developmental stages by MALDI-TOF MS profiling. Parasit Vectors 7: 544. s13071-014-0544-0 [pii]; doi: 10.1186/s13071-014-0544-0. pmid:25442218
[19]
Yssouf A, Parola P, Lindstrom A, Lilja T, L'Ambert G, Bondesson U, Berenger JM, Raoult D, Almeras L (2014) Identification of European mosquito species by MALDI-TOF MS. Parasitol Res 113: 2375–2378. doi: 10.1007/s00436-014-3876-y. pmid:24737398
[20]
Hoppenheit A, Murugaiyan J, Bauer B, Steuber S, Clausen PH, Roesler U (2013) Identification of Tsetse (Glossina spp.) using matrix-assisted laser desorption/ionisation time of flight mass spectrometry. PLoS Negl Trop Dis 7: e2305. doi: 10.1371/journal.pntd.0002305. PNTD-D-13-00052 [pii]. pmid:23875040
[21]
Yssouf A, Socolovschi C, Leulmi H, Kernif T, Bitam I, Audoly G, Almeras L, Raoult D, Parola P (2014) Identification of flea species using MALDI-TOF/MS. Comp Immunol Microbiol Infect Dis 37: 153–157. S0147-9571(14)00022-8 [pii]; doi: 10.1016/j.cimid.2014.05.002. pmid:24878069
[22]
Kaufmann C, Schaffner F, Ziegler D, Pfluger V, Mathis A (2012) Identification of field-caught Culicoides biting midges using matrix-assisted laser desorption/ionization time of flight mass spectrometry. Parasitology 139: 248–258. S0031182011001764 [pii]; doi: 10.1017/S0031182011001764. pmid:22008297
[23]
Sambou M, Aubadie-Ladrix M, Fenollar F, Fall B, Bassene H, Almeras L, Sambe-Ba B, Perrot N, Chatellier S, Faye N, Parola P, Wade B, Raoult D, Mediannikov O (2015) Comparison of matrix-assisted laser desorption ionization-time of flight mass spectrometry and molecular biology techniques for identification of Culicoides (Diptera: ceratopogonidae) biting midges in senegal. J Clin Microbiol 53: 410–418. JCM.01855-14 [pii]; doi: 10.1128/JCM.01855-14. pmid:25411169
[24]
Mathis A, Depaquit J, Dvorak V, Tuten H, Banuls AL, Halada P, Zapata S, Lehrter V, Hlavackova K, Prudhomme J, Volf P, Sereno D, Kaufmann C, Pfluger V, Schaffner F (2015) Identification of phlebotomine sand flies using one MALDI-TOF MS reference database and two mass spectrometer systems. Parasit Vectors 8: 266. doi: 10.1186/s13071-015-0878-2. 10.1186/s13071-015-0878-2 [pii]. pmid:25957576
[25]
Berdjane-Brouk Z, Charrel RN, Bitam I, Hamrioui B, Izri A (2011) Record of Phlebotomus (Transphlebotomus) mascittii Grassi, 1908 and Phlebotomus (Larroussius) chadlii Rioux, Juminer & Gibily, 1966 female in Algeria. Parasite 18: 337–339. P3V184R8 [pii]. pmid:22091465 doi: 10.1051/parasite/2011184337
[26]
Bounamous A, Boudabous R, Jouet D, Augot D, Ferte H, Babba H, Berchi S, Depaquit J (2008) [Molecular and morphological characterisation of two closely related species belonging to the subgenus Phlebotomus chabaudi Croset, Abonnenc & Rioux, 1970 et P. riouxi Depaquit, Killick-Kendrick & Leger, 1998 (Diptera: Psychodidae)]. Parasite 15: 565–571. pmid:19202763 doi: 10.1051/parasite/2008154565
[27]
Belazzoug S (1991) The sandflies of Algeria. Parassitologia 33 Suppl: 85–87. pmid:1841261
[28]
Izri MA, Belazzoug S, Pratlong F, Rioux JA (1992) [Isolation of Leishmania major in Phlebotomus papatasi in Biskra (Algeria). The end of an ecoepidemiological saga]. Ann Parasitol Hum Comp 67: 31–32. pmid:1642393
[29]
Boubidi SC, Benallal K, Boudrissa A, Bouiba L, Bouchareb B, Garni R, Bouratbine A, Ravel C, Dvorak V, Votypka J, Volf P, Harrat Z (2011) Phlebotomus sergenti (Parrot, 1917) identified as Leishmania killicki host in Ghardaia, south Algeria. Microbes Infect 13: 691–696. S1286-4579(11)00070-0 [pii]; doi: 10.1016/j.micinf.2011.02.008. pmid:21382502
[30]
Mihoubi I, Ramli I, Frahtia K, Kabouche Z, Harrat Z (2013) [Demonstration of the Leishmanicidal activity of Stachys guyoniana against Leishmania major in eastern Algeria]. Med Sante Trop 23: 230–231. mst.2013.0210 [pii]; doi: 10.1684/mst.2013.0210.
[31]
Adel A, Boughoufalah A, Saegerman C, De DR, Bouchene Z, Soukehal A, Berkvens D, Boelaert M (2014) Epidemiology of visceral leishmaniasis in Algeria: an update. PLoS One 9: e99207. doi: 10.1371/journal.pone.0099207. PONE-D-13-52034 [pii]. pmid:24949958
[32]
Leger N, Pesson B, Madulo-Leblond G, Abonnenc E (1983) [Differentiation of females of the subgenus Larroussius Nitzulescu 1931 (Diptera-Phlebotomidae) of the Mediterranean region]. Ann Parasitol Hum Comp 58: 611–623. pmid:6673646
[33]
Abonnenc E, Pastre J (1972) [Capture of Phlebotomus in the Republic of South Africa, with a description of P. macintoshi, n. sp. (Diptera, Psychodidae)]. Bull Soc Pathol Exot Filiales 65: 721–725. pmid:4679211
[34]
Depaquit J, Leger N, Killick-Kendrick R (1998) [Description of Phlebotomus (Paraphlebotomus) riouxi n. sp. (Diptera:Psychodidae) of northern Africa]. Parasite 5: 151–158. pmid:9754311 doi: 10.1051/parasite/1998052151
[35]
Wenk CE, Kaufmann C, Schaffner F, Mathis A (2012) Molecular characterization of Swiss Ceratopogonidae (Diptera) and evaluation of real-time PCR assays for the identification of Culicoides biting midges. Vet Parasitol 184: 258–266. S0304-4017(11)00598-X [pii]; doi: 10.1016/j.vetpar.2011.08.034. pmid:21963364
[36]
Depaquit J, Naucke TJ, Schmitt C, Ferte H, Leger N (2005) A molecular analysis of the subgenus Transphlebotomus Artemiev, 1984 (Phlebotomus, Diptera, Psychodidae) inferred from ND4 mtDNA with new northern records of Phlebotomus mascittii Grassi, 1908. Parasitol Res 95: 113–116. doi: 10.1007/s00436-004-1254-x. pmid:15666186
[37]
Asato Y, Oshiro M, Myint CK, Yamamoto Y, Kato H, Marco JD, Mimori T, Gomez EA, Hashiguchi Y, Uezato H (2009) Phylogenic analysis of the genus Leishmania by cytochrome b gene sequencing. Exp Parasitol 121: 352–361. S0014-4894(08)00339-1 [pii]; doi: 10.1016/j.exppara.2008.12.013. pmid:19159626
[38]
Saar Y, Ransford A, Waldman E, Mazareb S, Amin-Spector S, Plumblee J, Turco SJ, Zilberstein D (1998) Characterization of developmentally-regulated activities in axenic amastigotes of Leishmania donovani. Mol Biochem Parasitol 95: 9–20. S0166-6851(98)00062-0 [pii]. pmid:9763285 doi: 10.1016/s0166-6851(98)00062-0
[39]
Goyard S, Segawa H, Gordon J, Showalter M, Duncan R, Turco SJ, Beverley SM (2003) An in vitro system for developmental and genetic studies of Leishmania donovani phosphoglycans. Mol Biochem Parasitol 130: 31–42. S0166685103001427 [pii]. pmid:14550894 doi: 10.1016/s0166-6851(03)00142-7
[40]
Yssouf A, Socolovschi C, Flaudrops C, Ndiath MO, Sougoufara S, Dehecq JS, Lacour G, Berenger JM, Sokhna CS, Raoult D, Parola P (2013) Matrix-assisted laser desorption ionization—time of flight mass spectrometry: an emerging tool for the rapid identification of mosquito vectors. PLoS One 8: e72380. doi: 10.1371/journal.pone.0072380. PONE-D-13-13385 [pii]. pmid:23977292
[41]
Yssouf A, Almeras L, Terras J, Socolovschi C, Raoult D, Parola P (2015) Detection of Rickettsia spp in ticks by MALDI-TOF MS. PLoS Negl Trop Dis 9: e0003473. doi: 10.1371/journal.pntd.0003473. PNTD-D-14-01242 [pii]. pmid:25659152
[42]
Cherif K, Boudrissa A, Cherif MH, Harrat Z (2012) [A social program for the control of zoonotic cutaneous leishmaniasis in M'Sila, Algeria]. Sante Publique 24: 511–522. pmid:23473045
[43]
Yssouf A, Almeras L, Berenger JM, Laroche M, Raoult D, Parola P (2015) Identification of tick species and disseminate pathogen using hemolymph by MALDI-TOF MS. Ticks Tick Borne Dis. S1877-959X(15)00081-3 [pii]; doi: 10.1016/j.ttbdis.2015.04.013.
[44]
Kaufmann C, Ziegler D, Schaffner F, Carpenter S, Pfluger V, Mathis A (2011) Evaluation of matrix-assisted laser desorption/ionization time of flight mass spectrometry for characterization of Culicoides nubeculosus biting midges. Med Vet Entomol 25: 32–38. doi: 10.1111/j.1365-2915.2010.00927.x. pmid:21118284
[45]
Feltens R, Gorner R, Kalkhof S, Groger-Arndt H, von BM (2010) Discrimination of different species from the genus Drosophila by intact protein profiling using matrix-assisted laser desorption ionization mass spectrometry. BMC Evol Biol 10: 95. 1471-2148-10-95 [pii]; doi: 10.1186/1471-2148-10-95. pmid:20374617
[46]
Bond JG, Casas-Martinez M, Quiroz-Martinez H, Novelo-Gutierrez R, Marina CF, Ulloa A, Orozco-Bonilla A, Munoz M, Williams T (2014) Diversity of mosquitoes and the aquatic insects associated with their oviposition sites along the Pacific coast of Mexico. Parasit Vectors 7: 41. 1756-3305-7-41 [pii]; doi: 10.1186/1756-3305-7-41. pmid:24450800
[47]
Dantas-Torres F, Tarallo VD, Latrofa MS, Falchi A, Lia RP, Otranto D (2014) Ecology of phlebotomine sand flies and Leishmania infantum infection in a rural area of southern Italy. Acta Trop 137: 67–73. S0001-706X(14)00165-X [pii]; doi: 10.1016/j.actatropica.2014.04.034. pmid:24813871
[48]
Maia C, Parreira R, Cristovao JM, Freitas FB, Afonso MO, Campino L (2015) Molecular detection of Leishmania DNA and identification of blood meals in wild caught phlebotomine sand flies (Diptera: Psychodidae) from southern Portugal. Parasit Vectors 8: 173. doi: 10.1186/s13071-015-0787-4. 10.1186/s13071-015-0787-4 [pii]. pmid:25889732
[49]
Berdjane-Brouk Z, Charrel RN, Hamrioui B, Izri A (2012) First detection of Leishmania infantum DNA in Phlebotomus longicuspis Nitzulescu, 1930 from visceral leishmaniasis endemic focus in Algeria. Parasitol Res 111: 419–422. doi: 10.1007/s00436-012-2858-1. pmid:22350715
[50]
Maroli M, Cavallini C, Khoury C, Miceli N, Manilla G (1991) [Entomological survey of sandflies (Diptera, Psychodidae) in the province of L'Aquila (Abruzzo)]. Parassitologia 33: 127–131.
[51]
Bongiorno G, Habluetzel A, Khoury C, Maroli M (2003) Host preferences of phlebotomine sand flies at a hypoendemic focus of canine leishmaniasis in central Italy. Acta Trop 88: 109–116. S0001706X03001906 [pii]. pmid:14516922 doi: 10.1016/s0001-706x(03)00190-6
[52]
Tabbabi A, Ghrab J, Aoun K, Ready PD, Bouratbine A (2011) Habitats of the sandfly vectors of Leishmania tropica and L. major in a mixed focus of cutaneous leishmaniasis in southeast Tunisia. Acta Trop 119: 131–137. S0001-706X(11)00142-2 [pii]; doi: 10.1016/j.actatropica.2011.05.002. pmid:21605538
[53]
Pesson B, Ready JS, Benabdennbi I, Martin-Sanchez J, Esseghir S, Cadi-Soussi M, Morillas-Marquez F, Ready PD (2004) Sandflies of the Phlebotomus perniciosus complex: mitochondrial introgression and a new sibling species of P. longicuspis in the Moroccan Rif. Med Vet Entomol 18: 25–37. 471 [pii]. pmid:15009443 doi: 10.1111/j.0269-283x.2004.0471.x
[54]
Boudabous R, Jaouadi K, Bounamous A, Babba H (2012) Morphological and molecular investigations of population structure of Phlebotomus perniciosus and Phlebotomus longicuspis (Diptera: Psychodidae) in Tunisia. J Med Entomol 49: 787–793. pmid:22897038 doi: 10.1603/me11110
[55]
Berchi S, Bounamous A, Louadi K, Pesson B (2007) Differenciation morphologique de deux especes sympatriques: Phlebotomus perniciosus Newstead, 1911 et P. longicuspis Nitzulescu, 1930 (Diptera: Psychodidae). Ann. Soc. Entomol. Fr. 43: 201–203. doi: 10.1080/00379271.2007.10697511
[56]
Fotso FA, Mediannikov O, Diatta G, Almeras L, Flaudrops C, Parola P, Drancourt M (2014) MALDI-TOF mass spectrometry detection of pathogens in vectors: the Borrelia crocidurae/Ornithodoros sonrai paradigm. PLoS Negl Trop Dis 8: e2984. doi: 10.1371/journal.pntd.0002984. PNTD-D-14-00185 [pii]. pmid:25058611
