Parasite-mediated interactions within the insect vector: Trypanosoma rangeli strategies
© Garcia et al.; licensee BioMed Central Ltd. 2012
Received: 27 February 2012
Accepted: 30 May 2012
Published: 30 May 2012
Trypanosoma rangeli is a protozoan that is non-pathogenic for humans and other mammals but causes pathology in the genus Rhodnius. T. rangeli and R. prolixus is an excellent model for studying the parasite-vector interaction, but its cycle in invertebrates remains unclear. The vector becomes infected on ingesting blood containing parasites, which subsequently develop in the gut, hemolymph and salivary glands producing short and large epimastigotes and metacyclic trypomastigotes, which are the infective forms. The importance of the T. rangeli cycle is the flagellate penetration into the gut cells and invasion of the salivary glands. The establishment of the parasite depends on the alteration of some vector defense mechanisms. Herein, we present our understanding of T. rangeli infection on the vector physiology, including gut and salivary gland invasions, hemolymph reactions and behavior alteration.
KeywordsTrypanosoma rangeli Rhodnius prolixus Parasite and vector interactions Parasite development
Trypanosoma rangeli biological cycle
The life cycle of T. rangeli, which it shares in invertebrate and vertebrate hosts, is complex and mediated by numerous factors, which are still poorly understood [1–4]. Infective parasites have been found mainly in the salivary glands of R. prolixus[1–4]; although T. rangeli has also been found in the salivary glands of Triatoma dimiculata in Colombia . In the invertebrate host, the T. rangeli life cycle is characterized in three different regions of the insect vector: the gut, hemolymph and salivary glands, all of which are important for parasite development. The T. rangeli interactions in the vector begin with the ingestion of the trypomastigote forms in an infective bloodmeal. The parasites reach the gut of the insect vector and remain in the blood meal for some time after ingestion, but later they transform into epimastigotes that are able to multiply, and then normally cross the intestinal epithelium by an intracellular route and reach the hemocoel [1–4]. Then T. rangeli continues multiplying freely in the hemolymph or within hemocytes [6, 7], although T. rangeli can be destroyed by plasmatocytes . Thereafter, the flagellatesinvade the salivary glands, where they again multiply, and finally transform into metacyclic trypomastigotes, the forms that can be transmitted to mammalian hosts during a blood meal through salivary secretion [1–4, 8–10].
Although R. prolixus has an efficient system to eliminate pathogenic microorganisms, T. rangeli has the ability to survive in the hemolymph of R. prolixus counteracting the defense responses in many ways, and reaching the salivary glands to complete its life cycle in the invertebrate host [1–4, 6, 17–19]. In vivo and in vitro experiments have shown that oral infection with T. rangeli followed by inoculation of the insects with the same parasite inhibits hemocyte microaggregation reactions and release of arachidonic acid into the hemolymph of R. prolixus[18, 19]. Additionally, a T. rangeli oral infection significantly reduces the phagocytic activities of R. prolixus hemocytes by inhibition of the PAF and eicosanoids pathways . The mechanisms of this inhibition process is unknown, but some studies suggest that nitric oxide and superoxide could be the signaling molecules responsible to take the message from the gut to the hemocoel regulating anti-parasite reactions in this region [21, 22].
Some studies have revealed the presence of the short epimastigote forms of T. rangeli in the hemolymph during the first hours after hemocoel infection (Figure 2). After this period they transform into long epimastigote forms that are able to invade the hemocytes as well as the salivary glands [25, 26] (Figure 2). The inoculation of the short epimastigote forms was able to activate the R. prolixus proPO system in the hemolymph while the long form was not . This can be explained in part by the presence of a galactose-binding lectin purified from R. prolixus hemolymph that affects the survival and motility of short but not long T. rangeli epimastigotes forms .
Rhodnius species and/or T. rangeli strains may both be considered key factors for completing the parasite development in the vector [32, 33]. Recent investigations have demonstrated that the DNA mini satellites of the parasite may be involved in these interactions. The incubation of T. rangeli strains with R. prolixus hemolymph indicates the presence of a trypanolytic activity which acts against a T. rangeli KP1- isolated from R. colombiensis, but has no lytic activity against the T. rangeli KP1+ strain from R. prolixus, both species were from Colombia. The survival of the latter strain suggests that hemolymph of R. prolixus seems to be a biological barrier which does not allow the development and transmission of KP1- [32, 33].
Invasion of salivary glands
R. prolixus salivary glands are highly glycosylated and most protozoans have glycosylated compounds, as lectins or lectin-like molecules and enzymes, to regulate the parasite adhesion or invasion to host cells . Since epimastigoteforms invade salivary glands, ecto-phosphatase activity of long epimastigote forms could be involved at the interaction sites of parasites and salivary glands with d-galactose and specific lectin-receptors . Knowledge about salivary gland structures facilitates studies on the role of surface molecules in the attachment/invasion process by T. rangeli. Rhodnius salivary glands have been shown to be rich in carbohydrate moieties on their surface, and present diverse lectin binding patterns with specific carbohydrate residues in the basal, muscle, and cell layers of the glands [38, 40]. The carbohydrates detected on the salivary gland surface were used to investigate the adhesion between T. rangeli and the R. prolixus salivary glands. Experiments in vitro on attachment inhibition assays using long epimastigotes (the invasion/adhesion forms) demonstrated that some carbohydrates used were capable of inhibiting the receptors on both the salivary glands and T. rangeli surfaces . R. prolixus salivary glands have several lectins which present surface-related sugars, and diverse carbohydrate residues are present in the basal lamina, muscle, and cell layers of the gland. Incubation of Con A reacted intensely with the whole salivary glands and with basal membrane in particular, indicating high concentrations of mannosyl residues [38, 40]. In vitro sugar inhibition assays have demonstrated that some sugars tested attached to the surface of R. prolixus salivary glands and/or T. rangeli and inhibited the adhesion of the long epimastigotes to the gland surface . The attachment inhibition tests, using parasites or salivary glands pre-treated with sugar revealed that the highest inhibitory activities were observed with N-acetyl-D-glucosamine, N-acetyl-D-galactosamine, and galactose. These molecules may serve as receptors by which long forms of T. rangeli epimastigotes attach to the salivary gland surface, prior to invasion [38, 40].
Saliva and alterations in behavior
To overcome vertebrate reactions that prevent blood loss, saliva of R. prolixus contains dozens of different compounds with antihemostatic action, such as anticoagulants, antiplatelet and anti-inflammatory activities, and vasodilator compounds. Many of these biologically active salivary proteins belong to the lipocalin protein family [41–43]. The ultimate effect of this salivary antihemostatic apparatus is faster feeding by R. prolixus by decreasing the time required by the insect to locate the skin blood vessels and sucking blood efficiently [9, 44]. The efficiency of salivary parasite transmission is increased by prolonged intradermal probing time on the host by infected insect vectors. R. prolixus infected with T. rangeli displays enhanced probing time, and that infection of the salivary glands affected the feeding behavior of the vectors increasing the number of intradermal piercings on a rabbit host reducing the ability to suck the blood when the insect is infected with the flagellate [9, 45]. The prolongation of the probing time and the reduction of blood ingested in infected insects were not correlated to either the health of the insects or a physical obstruction of the food channel by the parasites by demonstrating that infected bugs probed and fed normally on a membrane artificial feeder . Thus, it is probable that a T. rangeli infection causes salivary gland pathology that must contribute to transmission efficiency. In fact, saliva production was drastically reduced in insects with salivary glands infected with T. rangeli, as evaluated by plasma clotting time, apyrase activity, and NO-like compounds [9, 41–43]. Since T. rangeli may damage the salivary gland cells of R. prolixus, salivary gland lesions could cause a deficiency in the biosynthesis processes of saliva components . Thus, T. rangeli impairs R. prolixus salivary gland function, preventing full expression of its antihemostatic machinery. This led the insects to prolong the duration of intradermal probing, which favors T. rangeli transmission. Finally, T. rangeli infection in the hemolymph of R. prolixus leads to a delay in molting, alters insect movements and can increase mortality [46–48].
There is ongoing research in our laboratory on the direct and indirect interactions between T. rangeli and R. prolixus in the insect gut, hemolymph and salivary glands, especially on the parasites cell tissue invasion. The flagellate mechanisms of the inhibition of cellular and humoral immune reactions in this insect vector need further investigation. However, it is becoming clear that some aspects of these gut and salivary gland invasions as well as the hemolymphatic compounds involved in this interaction are known. However, ligand-molecule interactions and several other surface molecules required for invasion of the gut and salivary glands must be better characterized. Additionally, epithelial immune reactions against the parasite invasion are poorly understood. Now, the increased availability of Rhodnius and T. rangeli functional genome analyses in combination with new experimental models, including double-stranded RNA knockdown, RNAi screens, transcriptomic and proteomic approaches, transgenesis, paratransgenesis of the vector and/or parasite, will offer a powerful tool for elucidating these parasite – vector interactions. Understanding how parasites are recognized as non-self, and how they have developed in the vector and the transmission strategies, facilitates the description of the molecular parasite-vector interface, vector competency, and provides unique opportunities to investigate the role of T. rangeli in shaping R. prolixus reactions against the parasite.
This work was supported by grants from Conselho de Desenvolvimento Científico e Tecnológico (CNPq) to ESG and PA; Fundação Oswaldo Cruz (FIOCRUZ)/Papes project to PA; Fundação Carlos Chagas Filho de Amparo a Pesquisa do Estado do Rio de Janeiro (FAPERJ) to PA, and Instituto Nacional de Ciência e Tecnologia em Entomologia Molecular (INCT-EM-CNPq). ESG and PA are Senior Scientists from CNPq; DPC and MBF are Pos-doc Fellowships from Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) and CNPq, respectively.
- Azambuja P, Garcia ES: Trypanosoma rangeliinteractions within the vectorRhodnius prolixus– a mini review. Mem Inst Oswaldo Cruz. 2005, 100: 567-572.View ArticlePubMedGoogle Scholar
- Hoare CC: Trypanosomes of Mammals, A Zoological Monograph. 1972, Blackwell, Oxford, EdinburghGoogle Scholar
- Guhl F, Vallejo GA: Trypanosoma(Herpetosoma)rangeliTejera 1920: an updated review. Mem Inst Oswaldo Cruz. 2003, 98: 435-442. 10.1590/S0074-02762003000400001.View ArticlePubMedGoogle Scholar
- D´Alessandro A: Biology ofTrypanosoma(Herpetosoma)rangeliTejera. Biology of Kinetoplastida, vol. 1. Edited by: Lamsden VHE, Evans DA. 1976, Academic, London, 327-493.Google Scholar
- Marinkelle CJ: Triatoma dimidiata capitata, a natural vector ofTrypanosoma rangeliin Colombia. Rev Biol Trop. 1968, 15: 203-205.Google Scholar
- Watkins R: Histology ofRhodnius prolixusinfected withTrypanosoma rangeli. J Invertebr Pathol. 1971, 17: 59-66. 10.1016/0022-2011(71)90126-1.View ArticlePubMedGoogle Scholar
- Oliveira MA, de Souza W: Further morphological studies on the behavior ofTrypanosoma rangeliin the hemocytes ofRhodnius prolixus. Parasitol Int. 2003, 52: 299-307. 10.1016/j.parint.2003.08.002.View ArticlePubMedGoogle Scholar
- Hecker H, Schwarzenbach M, Rudin W: Development and interactions ofTrypanosoma rangeliin and with the reduviid bug,Rhodnius prolixus. Parasitol Res. 1990, 76: 311-318. 10.1007/BF00928185.View ArticlePubMedGoogle Scholar
- Garcia ES, Mello CB, Azambuja P, Ribeiro JMC: Rhodnius prolixussalivary anti-hemostatic components decrease withTrypanosoma rangeliinfection. Exp Parasitol. 1994, 78: 287-293. 10.1006/expr.1994.1030.View ArticlePubMedGoogle Scholar
- Ferreira LL, Lorenzo MG, Elliot SL, Guarniere AA: A standardizable protocol for infection ofRhodnius prolixuswithTrypanosoma rangeli, which mimics natural infections and reveals physiological effects of infection upon the insect. J Invertebr Pathol. 2010, 105: 91-97. 10.1016/j.jip.2010.05.013.View ArticlePubMedGoogle Scholar
- Castro DP, Moraes C, Garcia ES, Azambuja P: Inhibitory effects of D-mannose on trypanosomatid lysis induced bySerratia marcescens. Exp Parasitol. 2007, 115: 200-204. 10.1016/j.exppara.2006.08.001.View ArticlePubMedGoogle Scholar
- Castro DP, Seabra S, Garcia ES, de Souza W, Azambuja P: Trypanosoma cruzi: ultrastructural studies of adhesion, lysis and biofilm formation bySerratia marcescens. Exp Parasitol. 2007, 117: 201-207. 10.1016/j.exppara.2007.04.014.View ArticlePubMedGoogle Scholar
- Eicher S, Schaub GA: Development of symbionts in triatomine bugs and the effects of infections with trypanososmatids. Exp Parasitol. 2002, 100: 17-27. 10.1006/expr.2001.4653.View ArticleGoogle Scholar
- Gomes AS, Graciano G, Nogueira NFS, de Souza W, Garcia ES, Azambuja P: Effects of gamma irradiation on the development ofTrypanosoma rangeliin the vectorRhodnius prolixus. J Invertebr Pathol. 2002, 79: 86-92. 10.1016/S0022-2011(02)00011-3.View ArticlePubMedGoogle Scholar
- Oliveira MA, de Souza W: An electron microscopic study of penetration byTrypanosoma rangeliinto mid-gut cells ofRhodnius prolixus. J Invertebr Pathol. 2001, 77: 22-26. 10.1006/jipa.2000.4988.View ArticlePubMedGoogle Scholar
- Machado PE, Eger-Mangrich I, Rosa G, Koerich LB, Grisard EC, Steindel M: Differential susceptibility of triatomines of the genusRhodniustoTrypanosoma rangelistrains from different geographical origins. Int J Parasitol. 2001, 31: 632-634. 10.1016/S0020-7519(01)00150-3.View ArticlePubMedGoogle Scholar
- Garcia ES, Castro DP, Figueiredo MB, Genta FA, Azambuja P: Trypanosoma rangeli: a new perspective for studying the modulation of immune reactions ofRhodnius prolixus. Parasit Vectors. 2009, 17: 33-View ArticleGoogle Scholar
- Garcia ES, Machado EMM, Azambuja P: Effects of eicosanoids biosynthesis inhibitors on the prophenoloxidase-activating system and microaggregation reactions in the hemolymph ofRhodnius prolixusinfected withTrypanosoma rangeli. J Insect Physiol. 2004, 50: 157-165. 10.1016/j.jinsphys.2003.11.002.View ArticlePubMedGoogle Scholar
- Garcia ES, Machado EMM, Azambuja P: Inhibition of hemocyte microaggregation reactions inRhodnius prolixuslarvae orally infected withTrypanosoma rangeli. Exp Parasitol. 2004, 107: 31-38. 10.1016/j.exppara.2004.03.015.View ArticlePubMedGoogle Scholar
- Figueiredo MB, Genta FA, Garcia ES, Azambuja P: Lipid mediators and vector infection:Trypanosoma rangeliinhibitsRhodnius prolixushemocytes phagocytosis by modulation of phospholipase A2 and PAF-acetylhydrolase activities. J Insect Physiol. 2008, 54: 1528-1537. 10.1016/j.jinsphys.2008.08.013.View ArticlePubMedGoogle Scholar
- Whitten MMA, Mello CB, Gomes SA, Nigam Y, Azambuja P, Garcia ES, Ratcliffe NA: Role of superoxide and reactive nitrogen intermediates inRhodnius prolixus(Reduviidae)/Trypanosoma rangeliinteractions. Exp Parasitol. 2001, 98: 44-57. 10.1006/expr.2001.4615.View ArticlePubMedGoogle Scholar
- Whitten MMA, Sun F, Tew I, Schaub GA, Soukou C, Nappi A, Ratcliffe NA: Differential modulation ofRhodnius prolixusnitric oxide activities following challenge with nitric oxide activities following challenge withTrypanosoma rangeli,T. cruziand bacterial wall components. Insect Biochem Mol Biol. 2007, 37: 440-452. 10.1016/j.ibmb.2007.02.001.View ArticlePubMedGoogle Scholar
- Gregorio EA, Ratcliffe NA: The phenoloxidase system and in vitro interaction ofTrypanosoma rangeliwithRhodnius prolixusandTriatoma infestanshaemolymph. Parasite Immunol. 1991, 13: 551-564. 10.1111/j.1365-3024.1991.tb00551.x.View ArticlePubMedGoogle Scholar
- Gregorio EA, Ratcliffe NA: The distribution of agglutinins and lytic activity againstTrypanosoma rangeliand erythrocytes inRhodnius prolixusandTriatoma infestanstissue extracts and haemolymph. Mem Inst Oswaldo Cruz. 1991, 86: 181-186.View ArticlePubMedGoogle Scholar
- Mello CB, Garcia ES, Ratcliffe NA, Azambuja P: Trypanosoma cruziandTrypanosoma rangeli: interplay with hemolymph components ofRhodnius prolixus. J Invertebr Pathol. 1995, 65: 261-268. 10.1006/jipa.1995.1040.View ArticlePubMedGoogle Scholar
- Gomes SAO, Feder D, Thomas NE, Garcia ES, Azambuja P: Rhodnius prolixusinfected withTrypanosoma rangeli: in vivo and in vitro experiments. J Invertebr Pathol. 1999, 73: 289-293. 10.1006/jipa.1998.4836.View ArticlePubMedGoogle Scholar
- Gomes SAO, Feder D, Garcia ES, Azambuja P: Suppression of the propheloxidase system inRhodnius prolixusorally infected withTrypanosoma rangeli. J Insect Physiol. 2003, 49: 829-837. 10.1016/S0022-1910(03)00133-1.View ArticlePubMedGoogle Scholar
- Takle GB: Studies on the cellular immune response of insects toward the insect pathogenTrypanosoma rangeli. J Invertebr Pathol. 1988, 51: 64-72. 10.1016/0022-2011(88)90089-4.View ArticlePubMedGoogle Scholar
- Ratcliffe NA, Nigan Y, Mello CB, Garcia ES, Azambuja P: Trypanosoma cruziand erythrocyte agglutinins: a comparative study of occurrence and properties in the gut and hemolymph ofRhodnius prolixus. Exp Parasitol. 1996, 83: 83-93. 10.1006/expr.1996.0052.View ArticlePubMedGoogle Scholar
- Mello CB, Azambuja P, Garcia ES, Ratcliffe NA: Differential in vitro and in vivo behavior of three strains ofTrypanosoma cruziin the gut and hemolymph ofRhodnius prolixus. Exp Parasitol. 1996, 82: 112-121. 10.1006/expr.1996.0015.View ArticlePubMedGoogle Scholar
- Mello CB, Nigam Y, Garcia ES, Azambuja P, Newton RP, Ratcliffe NA: Studies on a haemolymph lectin isolated from Rhodnius prolixus and its interaction withTrypanosoma rangeli. Exp Parasitol. 1999, 91: 289-296. 10.1006/expr.1998.4385.View ArticlePubMedGoogle Scholar
- Vallejo GA, Guhl F, Carranza JC, Triana O, Péres G, Ortiz PA, Marín DH, Villa M, Suáres J, Sánches LP, Pulido X, Rodrígues LB, Lozano LE, Urrea DA, Rivera FA, Cuba CC, Clavijo JA: Interacción tripanosoma-vector-vertebrado y su relación con la sistemática y la epidemiologia de la tripanosomiasis americana. Biomedica. 2007, 27: 110-118.View ArticlePubMedGoogle Scholar
- Pulido XC, Pérez G, Vallejo GA: Preliminary characterization of aRhodnius prolixushemolymph trypanolytic protein, this being a determinant ofTrypanosoma rangeliKP1(+) and KP1(−) subpopulations vectorial ability. Mem Inst Oswaldo Cruz. 2008, 103: 172-179. 10.1590/S0074-02762008000200008.View ArticlePubMedGoogle Scholar
- Anhê AC, Azeredo-Oliveira MT: Cytochemical characterization ofTriatoma infestansandPanstrongylus megistussalivary gland cells (Hemiptera, Reduviidae, Triatominae). Micron. 2008, 39: 1126-1133. 10.1016/j.micron.2008.06.003.View ArticlePubMedGoogle Scholar
- Stark KR, James AJ: The salivary glands of disease vector. The Biology of Disease Vector. Edited by: Beaty BJ, Marguardt WC. 2008, University Press of Colorado, New York, 333-347.Google Scholar
- Ellis DS, Evans DA, Stamford S: The penetration of the salivary glands ofRhodnius prolixusbyTrypanosoma rangeli. Z Parasitk. 1980, 62: 75-84. 10.1007/BF00925368.View ArticleGoogle Scholar
- Meirelles RM, Henriques-Pons A, Soares MJ, Steindel M: Penetration of the salivary glands ofRhodnius domesticusNeiva & Pinto, 1923 (Hemiptera: Reduviidae) byTrypanosoma rangeliTejera, 1920 (Protozoa: Kinetoplastida). Parasitol Res. 2005, 97: 259-269. 10.1007/s00436-005-1433-4.View ArticlePubMedGoogle Scholar
- Ferguson MA: The surface glycoconjugates of trypanosomatids parasites. Phil. Trans. Royal Soc. London, Series B. 1997, 352: 1295-1302. 10.1098/rstb.1997.0113.View ArticleGoogle Scholar
- Gomes SA, de Souza AL Fonseca, Kiffer-Moreira T, Dick CF, Dos Santos AL, Meyer-Fernandes JR: Ecto-phosphatase activity on the external surface ofRhodnius prolixussalivary glands: modulation by carbohydrates andTrypanosoma rangeli. Acta Trop. 2008, 106: 137-142. 10.1016/j.actatropica.2008.02.008.View ArticlePubMedGoogle Scholar
- Basseri IF, Tew IF, Ratcliffe NA: Identification and distribution of carbohydrate moieties on the salivary glands ofRhodnius prolixusand their possible involvement in attachment/invasion byTrypanosoma rangeli. Exp Parasitol. 2002, 100: 116-234.View ArticleGoogle Scholar
- Ribeiro JMC: Role of saliva in blood-feeding by arthropods. Ann Rev Entomol. 1987, 32: 463-478. 10.1146/annurev.en.32.010187.002335.View ArticleGoogle Scholar
- Ribeiro JMC: Vector saliva and its role in parasite transmission. Exp Parasitol. 1989, 69: 104-106. 10.1016/0014-4894(89)90177-X.View ArticlePubMedGoogle Scholar
- Ribeiro JMC, Andersen J, Silva-Neto MAC, Pham VM, Garfield MK, Velenzuela JG: Exploring the sialome of the blood-sucking bugRhodnius prolixus. Insect Biochem Mol Biol. 2004, 34: 61-79. 10.1016/j.ibmb.2003.09.004.View ArticlePubMedGoogle Scholar
- Ribeiro JMC, Garcia ES: The salivary and crop apyrase activity inRhodnius prolixus. J Insect Physiol. 1980, 26: 303-307. 10.1016/0022-1910(80)90138-9.View ArticleGoogle Scholar
- Anez N, East JS: Studies inTrypanosoma rangeliTejera, 1920.II. Its effects on the feeding behavior in triatomine bugs. Acta Trop. 1984, 41: 93-95.PubMedGoogle Scholar
- D´Alessandro A, Saravia NG: Trypanosoma rangeli. Parasitic protozoa. Edited by: Kreir JP, Baker J. 1992, Academic, London, 1-54.Google Scholar
- Schaub GA: The effects of trypanosomatids on insects. Adv Parasitol. 1992, 31: 255-319.View ArticlePubMedGoogle Scholar
- Schaub GA: Pathogenicity of trypanosomatids on insects. Parasitol Today. 1994, 10: 463-468. 10.1016/0169-4758(94)90155-4.View ArticlePubMedGoogle Scholar
This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.