Skip to main content Skip to main navigation menu Skip to site footer
Responsive image
Issue: Vol. 5471 No. 5: 21 Jun. 2024
Type: Article
Published: 2024-06-21
DOI: 10.11646/zootaxa.5471.5.4
Page range: 572-584
Abstract views: 133
PDF downloaded: 13

Hidden life cycle stages of the trypanosomatid Blastocrithidia triatomae Cerisola et al. 1971 (Protista: Excavata: Kinetoplastea) unveiled by a new cultivation method

Laboratório Interdisciplinar de Vigilância Entomológica de Diptera e Hemiptera; Instituto Oswaldo Cruz/FIOCRUZ; Rio de Janeiro/ RJ; Brazil; Laboratório de Insetos Vetores; Universidade de Vassouras; Campus Maricá; Maricá/RJ; Brazil; Faculdade de Ciências Médicas de Maricá; Maricá/RJ; Brazil
Laboratório Interdisciplinar de Vigilância Entomológica de Diptera e Hemiptera; Instituto Oswaldo Cruz/FIOCRUZ; Rio de Janeiro/ RJ; Brazil; Laboratório de Protistologia; Universidade Federal do Rio de Janeiro; Rio de Janeiro/RJ; Brazil
Laboratório Interdisciplinar de Vigilância Entomológica de Diptera e Hemiptera; Instituto Oswaldo Cruz/FIOCRUZ; Rio de Janeiro/ RJ; Brazil
Programa de Pós-graduação em Ciências Veterinárias da Universidade Federal Rural do Rio de Janeiro; Seropédica/RJ; Brazil
Programa de Iniciação Científica da Universidade Iguaçu; Universidade Iguaçu; Nova Iguaçu/RJ; Brazil
Laboratório de Diagnóstico Molecular e Hematologia; Universidade Federal do Rio de Janeiro; Rio de Janeiro; Brazil
Laboratório de Biodiversidade de Vetores e Patógenos; Universidade Federal Fluminense; Niterói/RJ; Brazil
Laboratório Interdisciplinar de Vigilância Entomológica de Diptera e Hemiptera; Instituto Oswaldo Cruz/FIOCRUZ; Rio de Janeiro/ RJ; Brazil; Programa de Iniciação Científica da Universidade Iguaçu; Universidade Iguaçu; Nova Iguaçu/RJ; Brazil; Escritório Regional FIOCRUZ Piauí; Teresina/PI; Brazil
Laboratório de Biologia de Insetos; Universidade Federal Fluminense; Niterói/RJ; Brazil
Protist Cultivation Morphology Morphometry Ontogeny Protista

Abstract

Blastocrithidia triatomae is a monoxenic trypanosomatid parasite of triatomines, sharing the same insect vectors with Trypanosoma cruzi Chagas, 1909 and T. rangeli Tejera, 1920. It is known to cause a complex syndrome in insects which induces severe metabolic disorders and increasing in mortality rates. In the present study, we established an efficient serial axenic cultivation method for B. triatomae using a new medium, herein called MBT, which revealed previously unknown morphotypes, named microepimastigotes and sinemastigotes, not found in specific cell linage cultures. The results improve the knowledge on the life cycle of genus Blastocrithidia Laird 1959, its distinction from other trypanosomatids, such as T. cruzi and T. rangeli, and settle new ground for thorough exploration of morphological features and its application for trypanosomatid systematics.

 

References

  1. Ahuja, K., Vats, A., Beg, M.A., Kariyawasam, K.K.G.D.U.L., Chaudhury, A., Chatterjee, M., Karunaweera, N.D. & Selvapandiyan, A. (2019) High resolution melting based method for rapid discriminatory diagnosis of co-infecting Leptomonas seymouri in Leishmania donovani-induced leishmaniasis. Parasitology International, 75, 102047. https://doi.org/10.1016/j.parint.2019.102047
  2. Ambit, A., Woods, K.L., Cull, B., Coombs, G.H. & Mottram, J.C. (2011) Morphological events during the cell cycle of Leishmania major. Eukaryotic Cell, 10, 1429–1438. https://doi.org/10.1128/EC.05118-11
  3. Barazesh, A., Motazedian, M.H., Fouladvand, M., Hatam, G., Tajbakhsh, S., Ebrahimi, S. & Purkamal, D., (2019) Molecular Identification of Species Caused Cutaneous Leishmaniasis in Southern Zone of Iran. Journal of Arthropod-Borne Diseases, 13, 198–205. https://doi.org/10.18502/jad.v13i2.1246
  4. Barros, J.H.S., Fonseca, T.S., Macedo-Silva, R.M., Côrte-Real, S., Toma, H.K. & Madeira, M.F. (2014) Aflagellar epimastigote forms are found in axenic culture of Trypanosoma caninum. Acta Tropica, 137, 147–151. https://doi.org/10.1016/j.actatropica.2014.05.013
  5. Berg, C. (1879) Hemiptera Argentina. Ensayo de uma monografía de los Hemípteros-Heterópteros y Homópteros de la República Argentina. Anales de la Sociedad Científica Argentina, 7 & 8, 86–92 + 225–236 + 262–278 & 19–33.
  6. Caicedo, A.M., Gallego, G., Munoz, J.E., Suarez, H., Torres, A.G., Carvajal, H., Carvajal, F.C., Posso, A.M., Maslov, D. & Montoya-Lerma, J. (2011) Morphological and molecular description of Blastocrithidia cyrtomeni sp. nov. (Kinetoplastea: Trypanosomatidae) associated with Cyrtomenus bergi Froeschner (Hemiptera: Cydnidae) from Colombia. Memórias do Instituto Oswaldo Cruz, 106, 301–307. https://doi.org/10.1590/S0074-02762011000300008
  7. Carvalho, A.L.M. (1973) Estudos sobre a posição sistemática, a biologia e a transmissão de tripanosomatídeos encontrados em Zelus leucogrammus (Perty, 1834) (Hemiptera, Reduviidae). Revista de Patologia Tropical, 2, 223–274.
  8. Cerisola, J.A., Del Prado, C.E., Rohwedder, R. & Bozzini, J.P. (1971) Blastocrithidia triatomae n. sp. found in Triatoma infestans from Argentina. Journal of Protozoology, 18, 503–506. https://doi.org/10.1111/j.1550-7408.1971.tb03362.x
  9. Chagas, C. (1909) Neue Trypanosomen: Vorläufige mitteilung. Archiv für Schiffs- und Tropen-Hygiene, Leipzig, 13, 120–122.
  10. D’Alessandro, A. (1976) Biology of Trypanosoma (Herpetosoma) rangeli Tejera, 1920. In: Lumsden, W.H. R. & Evans, D.A. (Eds.), Biology of the Kinetoplastida. Academic Press, London, pp. 327–403.
  11. Dedet, J.P., Melogno, R., Cardenas, F., Valda, L., David, C., Fernandez, V., Torrez, M. E., Dimier-David, L., Lyevre, P. & Villareal, M.E. (1995) Rural campaign to diagnose and treat mucocutaneous leishmaniasis in Bolivia. Bulletin of the World Health Organization, 73, 339–345.
  12. de Castro, L.A., Küppers, G.C., Fernandes, N.M., Schlegel, M. & Paiva, T. da S. (2016) Ontogeny and molecular phylogeny of Apoamphisiella vernalis reveal unclear separation between genera Apoamphisiella and Paraurostyla (Protozoa, Ciliophora, Hypotricha). PLoS ONE, 11, e0155825. https://doi.org/10.1371/journal.pone.0155825
  13. de Castro, L.A.M., da Silva-Neto, I.D. & Paiva, T. da S. (2016) Physiological reorganization in the hypotrich ciliate Apoamphisiella vernalis (Protista, Ciliophora, Hypotricha). Iheringia, Série Zoologia, 107, e2017012. https://doi.org/10.1590/1678-4766e2017012
  14. Devera, R., Fernandes, O. & Coura, J.R. (2003) Should Trypanosoma cruzi be called “cruzi” complex? A review of the parasite diversity and the potential of selecting population after in Vitro culturing and mice infection. Memórias do Instituto Oswaldo Cruz, 98, 1–12. https://doi.org/10.1590/S0074-02762003000100001
  15. Eigner, P. & Foissner, W. (1994) Divisional morphogenesis in Amphisiellides illuvialis n. sp., Paramphisiella caudata (Hemberger) and Hemiamphisiella terricola Foissner, and redefinition of the Amphisiellidae (Ciliophora, Hypotrichida). Journal of Eukaryotic Microbiology, 41, 243–261. https://doi.org/10.1111/j.1550-7408.1994.tb01505.x
  16. Frolov, A.O., Malysheva, M.N., Ganyukova, A.I., Yurchenko, V. & Kostygov, A.Y. (2017) Life cycle of Blastocrithidia papi sp. n. (Kinetoplastea, Trypanosomatidae) in Pyrrhocoris apterus (Hemiptera, Pyrrhocoridae). European Journal of Protistology, 57, 85–98. https://doi.org/10.1016/j.ejop.2016.10.007
  17. Frolov, A.O., Malysheva, M.N., Ganyukova, A.I., Spodareva, V.V., Králová, J., Yurchenko, V. & Kostygov, A.Y. (2020) If host is refractory, insistent parasite goes berserk: Trypanosomatid Blastocrithidia raabei in the dock bug Coreus marginatus. PLoS ONE, 15, e0227832. https://doi.org/10.1371/journal.pone.0227832
  18. Ghobakhloo, N., Motazedian, M.H., Naderi, S. & Ebrahimi, S. (2019) Isolation of Crithidia spp. from lesions of immunocompetent patients with suspected cutaneous leishmaniasis in Iran. Tropical Medicine & International Health, 24, 116–126. https://doi.org/10.1111/tmi.13042
  19. Goloboff, P.A. & Catalano, S.A. (2016) TNT version 1.5, including a full implementation of phylogenetic morphometrics. Cladistics, 32, 221–238. https://doi.org/10.1111/cla.12160
  20. Goloboff, P.A., Farris, J., Källersjö, M., Oxelmann, B., Ramírez, M. & Szumik, C. (2003) Improvements to resampling measures of group support. Cladistics, 19, 324–332. https://doi.org/10.1111/j.1096-0031.2003.tb00376.x
  21. Gruby, D. (1843) Recherches et observations sur une nouvelle espéce d’haematozoaire, Trypanosoma sanguinis. Comptes Rendus hebdomadaires des Séances de l’Académie des Sciences, 17, 1134–1136.
  22. Hausmann, K., Hülsmann, N. & Radek, R. (2003) Protistology. 3rd Edition. E. Schweizerbart’sche, Stuttgart, 379 pp.
  23. Hennig, W. (1966) Phylogenetic Systematics. University of Illinois Press, Urbana, Illinois, 263 pp.
  24. Hoare, C.A. (1964) Morphological and taxonomical studies on mammalian trypanosomes. X. Revision of the systematics. Journal of Protozoology, 11, 200–210. https://doi.org/10.1111/j.1550-7408.1964.tb01741.x
  25. Hoare, C.A. & Wallace, F.G. (1966) Developmental stages of trypanosomatid flagellates: A new terminology. Nature, 212, 1385–1386. https://doi.org/10.1038/2121385a0
  26. Hubsch, R., Nuñez, V., Mora, E. & Carrasquero, B. (1977) Blastocrithidia triatomae torrealbai n. subsp. encontrado en Triatoma maculata (Hemiptera, Reduviidae) de Venezuela. Boletin de la Dirección de Malariología y Saneamiento Ambiental, 17, 14–19.
  27. Kaufer, A., Ellis, J., Stark, D. & Barret J. (2017) The evolution of trypanosomatid taxonomy. Parasites & Vectors, 10, 287. https://doi.org/10.1186/s13071-017-2204-7
  28. Kitching, I.J., Culverwell, C.L. & Harbach, R.E. (2014) The phylogenetic conundrum of Lutzia (Diptera: Culicidae: Culicini): a cautionary account of conflict and support. Insect Systematics & Evolution, 46, 269–290. https://doi.org/10.1163/1876312X-45032119
  29. Klug, F. (1834) Reise um die Erde, in den Jahren 1830, 1831 und 1832 ausgeführt von F. J. F. Meyen. Teil 1. C. W. Eichoff, Berlin, 493 pp.
  30. Kollien, A.H. & Schaub, G.A. (2003) The development of Blastocrithidia triatomae (Trypanosomatidae) in the reduviid bug Triatoma infestans (Insecta): influence of feeding. Parasitology Research, 89, 430–436. https://doi.org/10.1007/s00436-002-0790-5
  31. Laird, M. (1959) Blastocrithidia n.g. (Mastigophora: Protomonadina) for Crithidia (in part), with a subarctic record for B. gerridis (Patton). Canadian Journal of Zoology, 37, 749–752. https://doi.org/10.1139/z59-075
  32. Laporte, F.L. (1832) Essai d’une classification systématique de l’ordre des Hémiptères (Hémiptères Hétéroptères, Latr.). Magazine de Zoologie, Gudrin, 2, 88 pp., pls. 51–55.
  33. Léger, L. (1902) Sur un flagellé parasite de l’Anopheles maculipennis. Comptes rendus hebdomadaires des séances et mémoires de la Société de biologie, 54, 354–356.
  34. Lipa, J.J. (1968) Some observations on flagellate parasites of hemipterans Corimelaena, Euschistus, Gerris, Leptocoris and Oncopeltus in the United States. Acta Protozoologica, 6, 58–69.
  35. Lwoff, M. & Lwoff, A. (1931) Recherches sur la morphologie de Leptomonas oncopelti Noguchi et Tilden et Leptomonas fasciculata Novy, McNeal et Torrey. Archives de zoologie expérimentale et Générale, 71, 21–37.
  36. Maruyama, S.R., Santana, A., Takamiya, N.T., Takahashi, T.Y., Rogerio, L.A., Oliveira, C., Milanezi, C.M., Trombela, V.A., Cruz, A.K., Jesus, A.R., Barreto, A.S., Silva, A.M., Almeida, R.P., Ribeiro, J.M. & Silva, J.S. (2019) Non-Leishmania Parasite in Fatal Visceral Leishmaniasis–Like Disease, Brazil. Emerging Infectious Diseases, 25, 2088–2092. https://doi.org/10.3201/eid2511.181548
  37. McCulloch, I. (1915). An outline of the morphology and life history of Crithidia leptocoridis, sp. nov. University of California Publications in Zoology, 16, 1–22.
  38. McGhee, R.B. & Hanson, W.L. (1964) Comparison of the life cycle of Leptomonas Oncopelti and Phytomonas elmassiani. Journal of Protozoology, 11, 555–562. https://doi.org/10.1111/j.1550-7408.1964.tb01798.x
  39. Novy, F.G., McNeal, W.J. & Torrey, H.N. (1907) The trypanosomes of mosquitoes and other insects. The Journal of Infectious Diseases, 4, 223–276. https://doi.org/10.1093/infdis/4.2.223
  40. Noyes, H., Pratlong, F., Chance, M., Ellis, J., Lanotte, G. & Dedet, J.P. (2002) A previously unclassified trypanosomatid responsible for human cutaneous lesions in Martinique (French West Indies) is the most divergent member of the genus Leishmania ss. Parasitology, 124, 17–24. https://doi.org/10.1017/S0031182001008927
  41. Oladiran, A. & Belosevic, M. (2012) Immune evasion strategies of trypanosomes: A review. Journal of Parasitology, 98, 284–292. https://doi.org/10.1645/GE-2925.1
  42. Pacheco, R.S., Ferreira, M.S., Machado, M.I., Brito, C.M., Pires, M.Q., Da-Cruz, A.M. & Coutinho, S.G., (1998) Chagas’ disease and HIV co-infection: genotypic characterization of the Trypanosoma cruzi strain. Memórias do Instituto Oswaldo Cruz, 93, 165–169. https://doi.org/10.1590/S0074-02761998000200005
  43. Patton, W.S. (1908) The life-cycle of a species of Crithidia parasitic in the intestinal tract of Gerris fossarum Fabr. Archiv für Protistenkunde, 12, 131–146.
  44. Peng, P.L.M. & Wallace, F.G. (1981) The cultivation of Blastocrithidia triatomae Cerisola et al. 1971. Journal of Protozoology, 28, 116–118. https://doi.org/10.1111/j.1550-7408.1981.tb02814.x
  45. Peng, P.L.M. & Wallace, F.G. (1982) The cysts of Blastocrithidia triatomae Cerisola et al. 1971. Journal of Eukaryotic Microbiology, 29, 464–467. https://doi.org/10.1111/j.1550-7408.1982.tb05434.x
  46. Perlowagora-Szumlewicz, A. & Moreira, C.J.C. (1994) In vivo differentiation of Trypanosoma cruzi. I. Experimental evidence of the influence of vector species on metacyclogenesis. Memórias do Instituto Oswaldo Cruz, 89, 603–618. https://doi.org/10.1590/S0074-02761994000400018
  47. Pudney, M. & Lanar, D. (1977) Establishment and characterization of a cell line (BTC-32) from the triatomine bug, Triatoma infestans (Klug) (Hemiptera: Reduviidae). Annals of tropical medicine and parasitology, 71, 109–117. https://doi.org/10.1080/00034983.1977.11687167
  48. Ranque, P., Nourrit, J. & Nicoli, R.M. (1974) Recherches systématiques sur les trypanosomides. Les stades évolutifs chez les trypanosomides. Bulletin de la Societe de Pathologie Exotique, 67, 377–387.
  49. Reduth, D., Schaub, G.A. & Pudney, M. (1989) Cultivation of Blastocrithidia triatomae (Trypanosomatidae) on a cell line of its host Triatoma infestans (Reduviidae). Parasitology, 98, 387–393. https://doi.org/10.1017/S0031182000061461
  50. Rocha-e-Silva, E.O., Pattoli, D.G.B., Correa, R.R. & Andrade, J.C.R. (1977) Observações sobre o encontro de tripanosomatídeos do gênero Blastocrithidia, infetando naturalmente triatomíneos em insetário e no campo. Revista de Saúde Pública, São Paulo, 11, 87–96. https://doi.org/10.1590/S0034-89101977000100008
  51. Rocha-Silva, E.O., Souza, J.M., Andrade, J.C., Mello, C.S. & Ferreira, O.A. (1977) Preferência alimentar (entre sangue humano e ave) dos Triatoma sordida encontrados em casas habitadas da região norte do Estado de São Paulo-Brasil. Revista de Saúde Pública, São Paulo, 11, 258–269. https://doi.org/10.1590/S0034-89101977000200010
  52. Romeiro, A., Solé-Cava, A., Sousa, M.A., De Souza, W. & Attias, M. (2000) Ultrastructural and biochemical characterization of promastigotes and cystic form of Leptomonas wallacei n. sp. isolated from the intestine of its natural host Oncopeltus fasciatus (Hemiptera: Lygaeidae). Journal of Eukaryotic Microbiology, 47, 208–220. https://doi.org/10.1111/j.1550-7408.2000.tb00040.x
  53. Schaub, G.A. (1988) Developmental time and mortality in larvae of the reduviid bugs Triatoma infestans and Rhodnius prolixus after coprophagic infection with Blastocrithidia triatomae (Trypanosomatidae). Journal of Invertebrate Pathology, 51, 23–31. https://doi.org/10.1016/0022-2011(88)90084-5
  54. Schaub, G.A. (1990) Membrane feeding for infection of the reduviid bug Triatoma infestans with Blastocrithidia triatomae (Trypanosomatidae) and pathogenic effects of the flagellate. Parasitology Research, 76, 306–310. https://doi.org/10.1007/BF00928184
  55. Schaub, G.A. & Jensen, C. (1990) Developmental time and mortality of the reduviid bug Triatoma infestans with differential exposure to coprophagic infections with Blastocrithidia triatomae (Trypanosomatidae). Journal of Invertebrate Pathology, 55, 17–27. https://doi.org/10.1016/0022-2011(90)90027-4
  56. Schaub, G.A. & Pretsch, M. (1981) Ultrastructural studies on the excystment of Blastocrithidia triatomae (Trypanosomatidae). Transactions of the Royal Society of Tropical Medicine and Hygiene, 75, 168–171. https://doi.org/10.1016/0035-9203(81)90057-2
  57. Schaub, G.A., Böker, C.A., Jensen, C. & Reduth, D. (1989) Cannibalism and coprophagy are modes of transmission of Blastocrithidia triatomae (Trypanosomatidae) between triatomines. Journal of Protozoology, 36, 171–175. https://doi.org/10.1111/j.1550-7408.1989.tb01067.x
  58. Schaub, G.A. & Breger, B. (1988) Pathological effects of Blastocrithidia triatomae (Trypanosomatidae) on the reduviid bugs Triatoma sordida, T. pallidipennis and Dipetalogaster maxima after coprophagic infection. Medical and veterinary entomology, 2, 309–318. https://doi.org/10.1111/j.1365-2915.1988.tb00202.x
  59. Silva-Júnior, R. da & Paiva, T. da S. (2018) Evaluating the role of morphological characters in the phylogeny of some trypanosomatid genera (Excavata, Kinetoplastea, Trypanosomatida). Cladistics, 34, 167–180. https://doi.org/10.1111/cla.12199
  60. Siqueira-Castro, I.C.V., Paiva, T.S. & Silva-Neto, I.D. (2009) Morphology of Parastrongylidium estevesi comb. nov. and Deviata brasiliensis sp. nov. (Ciliophora: Stichotrichia) from a sewage treatment plant in Rio de Janeiro, Brazil. Zoologia, 26, 774–786. https://doi.org/10.1590/S1984-46702009000400024
  61. Sousa M.A. (1994) Trypomastigotes in cultures of Blastocrithidia culicis (Novy, MacNeal & Torrey, 1907) (Kinetoplastida: Trypanosomatidae). Memórias do Instituto Oswaldo Cruz, 89, 111–112. https://doi.org/10.1590/S0074-02761994000100018
  62. Sousa, M.A. (1999) Morphobiological characterization of Trypanosoma cruzi Chagas, 1909 and its distinction from other trypanosomes. Memórias do Instituto Oswaldo Cruz, 94, 205–210. https://doi.org/10.1590/S0074-02761999000700031
  63. Sousa, M.A. (2000) Tripanosomatídeos de insetos e plantas: análise do crescimento, diferenciação e divisão celular, biometria e fenômenos sugestivos de sexualidade; valor taxonômico. Instituto Oswaldo Cruz, Rio de Janeiro, 161 pp.
  64. Sousa, M.A. & Côrte-Real, S. (1991) Postnuclear kinetoplast in choanomastigotes of Crithidia deanei Carvalho, 1973: Proposal of a new genus. Revista do Instituto de Medicina Tropical de São Paulo, 33 (Supplement), S8.
  65. Stål, C. (1859) Monographie der Gattung Conorhinus und Verwandten. Berliner entomologische Zeitschrift, 3, 99–117, pl. 6, figs. I–IV. https://doi.org/10.1002/mmnd.18590030202
  66. Takata, C.S.A., Camargo, E.P. & Milder, R.V. (1996) Encystment and excysment of a trypanosomatid of the genus Leptomonas. European Journal of Protistology, 32, 90–95. https://doi.org/10.1016/S0932-4739(96)80043-5
  67. Tamura, K., Stecher, G., Peterson, D., Filipski, A. & Kumar, S. (2013) MEGA6: Molecular Evolutionary Genetics Analysis Version 6.0. Molecular Biology and Evolution, 30, 2725–2729. https://doi.org/10.1093/molbev/mst197
  68. Tejera, E. (1920) Un nouveau flagellé de Rhodnius prolixus, Trypanosoma (ou Crithidia) rangeli, n.sp. Bulletin de la Société de Pathologie Exotique, 13, 527–530.
  69. Teixeira, M.M.G., Borghesan, T.C., Ferreira, R.C., Santos, M.A., Takata, C.S.A., Campaner, M., Nunes, V.L.B., Milder, R.V., de Souza, W. & Camargo, E.P. (2011) Phylogenetic validation of the genera Angomonas and Strigomonas of trypanosomatids harboring bacterial endosymbionts with the description of new species of trypanosomatids and of proteobacterial symbionts. Protist, 162, 503–524. https://doi.org/10.1016/j.protis.2011.01.001
  70. Usinger, R.L. (1939) n.k. In: Descriptions of new Triatominae with a key to genera (Hemiptera, Reduviidae). University of California publications in entomology. Vol. 7. University of California Press, Berkeley, California, pp. 33–56, pl. 1, 2 figs.
  71. Vallejo, G.A., Marinkelle, C.J., Guhl, F. & Sanchez, N. (1988) Behavior of the infection and morphologic differentiation of Trypanosoma cruzi and T.rangeli in the intestine of the vector Rhodnius prolixus. Revista Brasileira de Biologia, 48, 577–587.
  72. Vickerman, K. (1990) Diplomonadida. In: Margulis, L., Corliss, J.O., Melkonian, M. & Chapman D.J. (Eds.), Handbook of the Protoctista; the structure, cultivation, habits and life histories of the eukaryotic microorganisms and their descendants exclusive of animals, plants and fungi. Jones and Bartlett Publishers, Boston, Massachusetts, pp. 200–210.
  73. Wallace, F.G (1966) The Trypanosomatid parasites of insects and arachnids. Experimental Parasitology, 18, 124–193. https://doi.org/10.1016/0014-4894(66)90015-4
  74. Wallace, F.G. (1977) Developmental stages of trypanosomatid flagellates: a new terminology revisited. Protozoology, 3, 51–56.
  75. Wheeler, R.J., Gluenz, E. & Gull, K. (2013) The Limits on Trypanosomatid Morphological Diversity. PLoS ONE, 8, e79581. https://doi.org/10.1371/journal.pone.0079581