title: Comparative analysis of salivary gland transcriptomes of Phlebotomus orientalis sand flies from endemic and non-endemic foci of visceral leishmaniasis.
abstract: BACKGROUND
  In East Africa, Phlebotomus orientalis serves as the main vector of Leishmania donovani, the causative agent of visceral leishmaniasis (VL). Phlebotomus orientalis is present at two distant localities in Ethiopia; Addis Zemen where VL is endemic and Melka Werer where transmission of VL does not occur. To find out whether the difference in epidemiology of VL is due to distant compositions of P. orientalis saliva we established colonies from Addis Zemen and Melka Werer, analyzed and compared the transcriptomes, proteomes and enzymatic activity of the salivary glands.
  
  
  RESULTS
  Two cDNA libraries were constructed from the female salivary glands of P. orientalis from Addis Zemen and Melka Werer. Clones of each P. orientalis library were randomly selected, sequenced and analyzed. In P. orientalis transcriptomes, we identified members of 13 main protein families. Phylogenetic analysis and multiple sequence alignments were performed to evaluate differences between the P. orientalis colonies and to show the relationship with other sand fly species from the subgenus Larroussius. To further compare both colonies, we investigated the humoral antigenicity and cross-reactivity of the salivary proteins and the activity of salivary apyrase and hyaluronidase.
  
  
  CONCLUSIONS
  This is the first report of the salivary components of P. orientalis, an important vector sand fly. Our study expanded the knowledge of salivary gland compounds of sand fly species in the subgenus Larroussius. Based on the phylogenetic analysis, we showed that P. orientalis is closely related to Phlebotomus tobbi and Phlebotomus perniciosus, whereas Phlebotomus ariasi is evolutionarily more distinct species. We also demonstrated that there is no significant difference between the transcriptomes, proteomes or enzymatic properties of the salivary components of Addis Zemen (endemic area) and Melka Werer (non-endemic area) P. orientalis colonies. Thus, the different epidemiology of VL in these Ethiopian foci cannot be attributed to the salivary gland composition.
authors.affiliation: Department of Parasitology, Faculty of Science, Charles University, Prague, Czech Republic.
authors.forename: Michaela
authors.initials: M
authors.lastname: Vlkova
authors.affiliation: Department of Parasitology, Faculty of Science, Charles University, Prague, Czech Republic.
authors.forename: Michal
authors.initials: M
authors.lastname: Sima
authors.affiliation: Department of Parasitology, Faculty of Science, Charles University, Prague, Czech Republic.
authors.forename: Iva
authors.initials: I
authors.lastname: Rohousova
authors.affiliation: Department of Parasitology, Faculty of Science, Charles University, Prague, Czech Republic.
authors.forename: Tatiana
authors.initials: T
authors.lastname: Kostalova
authors.affiliation: Department of Parasitology, Faculty of Science, Charles University, Prague, Czech Republic.
authors.forename: Petra
authors.initials: P
authors.lastname: Sumova
authors.affiliation: Department of Parasitology, Faculty of Science, Charles University, Prague, Czech Republic.
authors.forename: Vera
authors.initials: V
authors.lastname: Volfova
authors.affiliation: Genomics Unit, Research Technologies Section, Rocky Mountain Laboratories, Hamilton, Montana, United States of America.
authors.forename: Erin L
authors.initials: EL
authors.lastname: Jaske
authors.affiliation: Genomics Unit, Research Technologies Section, Rocky Mountain Laboratories, Hamilton, Montana, United States of America.
authors.forename: Kent D
authors.initials: KD
authors.lastname: Barbian
authors.affiliation: Aklilu Lemma Institute of Pathobiology, Addis Ababa University, Addis Ababa, Ethiopia.
authors.forename: Teshome
authors.initials: T
authors.lastname: Gebre-Michael
authors.affiliation: Department of Microbiology, Immunology & Parasitology, Faculty of Medicine, Addis Ababa University, Addis Ababa, Ethiopia.
authors.forename: Asrat
authors.initials: A
authors.lastname: Hailu
authors.affiliation: Department of Parasitology, The Kuvin Centre for the Study of Infectious and Tropical Diseases, Hadassah Medical School, The Hebrew University of Jerusalem, Jerusalem, Israel.
authors.forename: Alon
authors.initials: A
authors.lastname: Warburg
authors.affiliation: Vector Biology Section, Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rockville, Maryland, United States of America.
authors.forename: Jose M C
authors.initials: JM
authors.lastname: Ribeiro
authors.affiliation: Vector Molecular Biology Section, Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rockville, Maryland, United States of America.
authors.forename: Jesus G
authors.initials: JG
authors.lastname: Valenzuela
authors.affiliation: Vector Molecular Biology Section, Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rockville, Maryland, United States of America.
authors.forename: Ryan C
authors.initials: RC
authors.lastname: Jochim
authors.affiliation: Department of Parasitology, Faculty of Science, Charles University, Prague, Czech Republic.
authors.forename: Petr
authors.initials: P
authors.lastname: Volf
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D004798:Enzymes / Q000737:chemistry / Q000145:classification / Q000235:genetics; D005002:Ethiopia; D005260:Female; D007303:Insect Vectors / Q000235:genetics*; D007898:Leishmaniasis, Visceral / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010691:Phlebotomus / Q000235:genetics*; D012469:Salivary Glands / Q000737:chemistry* / Q000201:enzymology; D012471:Salivary Proteins and Peptides / Q000737:chemistry / Q000145:classification / Q000235:genetics* / Q000276:immunology; D016415:Sequence Alignment; D059467:Transcriptome / Q000235:genetics*
chemical_list: D004798:Enzymes; D012471:Salivary Proteins and Peptides