SCHULTE zyxwvutsrqp & SCHOLZE-E. HISTOL YTICA ON EXTRACELLULAR MATRIX zyxw 543 15. Lushbaugh, W. B., Hofbauer, A. F. & Pittman, F. E. 1985. zyxwvuts Entamoeba histolytica: purification of cathepsin B. Exp. Parasitol., zyxwvut 59: 16. Martinez-Hernandez, A. & Amenta, P. zyxwvutsrq S. 1983. The basement membrane in pathology. Lab. Invest., 48:656-677. 17. McLaughlin, F. & Faubert, G. 1976. Partial purification and properties of a neutral sulfhydryl and an acid proteinase from Ent- amoeba histolytica. Can. J. Microbiol., 23:420-425. 18. Miller, E. J. & Rhodes, R. K. 1982. Preparation and charac- terization of the different types of collagen. Methods Enzymol., 82:33- 64. 19. Muiioz, M. D. L., Rojkind, M., Calderon, J., Tanimoto, M., Arias-Negrete, S. & Martinez-Palomo, A. 1984. Entamoeba histoly- tica: collagenolytic activity and virulence. J. Protozool., 31:468-470. 20. Neal, R. A. 1960. Enzymatic proteolysis by Entamoeba his- tolytica; biochemical characteristics and relationship with invasiveness. Parasitology, 50:53 1-550. 21. Niyibizi, C., Fietzek, P. P. & Rest, M. V. D. 1984. Human placenta type V collagens. J. Biol. Chem., 259:14170-14174. 22. Plques, E. P., Scholze, H. & Huber, R. 1980. Purification and crystallization of human anaphylatoxin, C3A. Hoppe-Seyler’s Z. Phys- iol. Chem., 361:977-980. 23. Piez, K. A. 1968. Molecular weight determination of random coil polypeptides from collagen by molecular sieve chromatography. Anal. Biochem., 26:3054 12. 32 8-3 36. 24. Sakashita, S., Engvall, E. & Ruoslahti, E. 1980. Basement mem- brane glycoprotein laminin binds to heparin. FEBS-Lett., 116:243-246. 25. Scholze, H. & Wemes, E. 1984. A weakly acidic protease has a powerful proteolytic activity in Entamoeba histolytica. Mol. Biochem. Parasitol., 11:293-300. 26. Scholze, H. & Schulte, W. 1988. On the specificity of a cysteine proteinase from Entamoeba histolytica. Biomed. Biochim. Acta, 47: 1 15- 123. 27. Schulte, W., Scholze, H. & Wemes, E. 1987. Specificity of a cysteine proteinase of Entamoeba histolytica towards the zyx a 1 -CB2 pep- tide of bovine collagen type I. Mol. Biochem. Parasitol., 2539-43. 28. Takeuchi, A. & Phillips, B. P. 1975. Electron microscope study of experimental Entamoeba histolytica infections in the guinea pig. I Penetration of the intestinal epithelium by trophozoites. J. Trop. Med. Hyg., 24:34-48. 29. Takio, K., Towatari, T., Katunuma, N., Teller, D. & Titani, K. 1983. Homology of amino acid sequences of rat liver cathepsins B and H with papain. Proc. Natl. Acad. Sci. USA, 00:3666-3670. 30. Talamas-Rohana, P. & Meza, I. 1988. Interaction between pathogenic amebas and fibronectin: substrate degradation and changes in cytoskeleton organization. J. Cell Biol., 106: 1787-1794. Received 1-17-89; accepted 7-14-89 J. Protozoo/.. 36(6), 1989, pp. zyxwvutsrqponm 543-547 zyxwvutsrqponm 0 I989 by the Society of Protozoologists Culture and Generic Identification of Trypanosomatids of Phytophagous Hemiptera in Brazil CLIZETE SBRAVATE,* MARTA CAMPANER,* LUIS E. A. CAMARGO,** IVETE CONCHON,* MARTA M. G. TEIXEIRA* and E. PLESSMANN CAMARGO* *Departamento de Parasitologia. Instituto de Ciincias Biomgdicas, Universidade de SZo Paulo, 05508, SZo Paulo, Brazil and **Departamento de Fitopatologia, ESALQ, 13400, Piracicaba, SZo Paulo, Brazil ABSTRACT. Among the 372 phytophagous Hemiptera examined, 133 insects of 28 species (Coreidae 18, Pentatomidae 7, brrho- coridae 2, Lygaeidae 1) were infected with trypanosomatids. Gut infections only were found in 68.4%, gut and salivary gland infections in 29.3% and salivary infections alone in 2.3%. Fifty-one cultures were isolated from 38 insects. Cultures were characterized by assay of certain ornithine-arginine metabolism enzymes and by indirect immunofluorescence against monoclonal antibodies specific for Phytomonas spp. Ten cultures were identified as either Crithidia or Leptomonas. Twenty-one promastigote cultures had an enzyme pattern hitherto recorded only for Leishmania and 16 cultures were identified as Phytomonas. Key words. Enzymes, monoclonal antibodies, Phytomonas, taxonomy. NSECT trypanosomatids of Crithidia, Leptomonas, Herpe- I tomonas, and Blastocrithidia are commonly found in the digestive tract of phytophagous and predaceous Hemiptera [ 151. In addition, phytophagous hemipterans may also harbor plant parasites ofthe genus Phytomonas which they presumably trans- mit through saliva to their host plants [5]. Some species of Phytomonas have unquestioned importance as plant pathogens, allegedly causing diseases in coffee, coconut, oil palms and cassava [6, 8, 10, 121. Phytophagous hemipterans being their putative vectors, it would be highly desirable to have a correct appraisal of the prevalence of Phytomonas in these insects. However this task is rendered difficult by the limitations of morphological criteria [ 161. Among the trypanosomatids of phytophagous hemipterans, Blastocrithidia spp. can be easily recognized because of their possession of an undulating membrane. In theory, Crithidia spp. could also be easily identified since they occur as genus-specific choanomastigotes. However, not all choanomastigotes are of the classical barleycorn-shaped type and it is common knowl- edge that environmental conditions may act to blur the dis- tinction between choano- and promastigotes. Flagellates of the 3 remaining genera- Leptomonas, Herpetomonas, and Phytom- onas-occur in insects as promastigotes and cannot be distin- guished morphologically except for the unpredictable occur- rence of opisthomastigotes in Herpetomonas. Therefore additional methods are necessary for proper identification and determination of the prevalence of Phytomonas among phy- tophagous hemipterans. Enzymes of ornithine-arginine metab- olism occur in genus-specific patterns among Trypanosomati- dae. Thus arginase, present in Crithidia and Leptomonas may serve to distinguish these genera from Herpetomonas and Phy- tomonas which lack the enzyme [2, zyxw 3,7, 171. Further distinction between Herpetomonas and Phytomonas spp. can be made by monoclonal antibodies specific for phytomonads [ 141. Joint uti- lization of these methods would permit identification of Phy- tomonas among trypanosomatids of phytophagous hempiterans