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Redalyc.Hemoparasites of the Genus Trypanosoma (Kinetoplastida Anais da Academia Brasileira de Ciências ISSN: 0001-3765 [email protected] Academia Brasileira de Ciências Brasil LEAL, DENISE D.M.; O'DWYER, LUCIA H.; RIBEIRO, VITOR C.; SILVA, REINALDO J.; FERREIRA, VANDA L.; RODRIGUES, ROZANGELA B. Hemoparasites of the genus Trypanosoma (Kinetoplastida: Trypanosomatidae) and hemogregarines in Anurans of the São Paulo and Mato Grosso do Sul States - Brazil Anais da Academia Brasileira de Ciências, vol. 81, núm. 2, junio, 2009, pp. 199-206 Academia Brasileira de Ciências Rio de Janeiro, Brasil Available in: http://www.redalyc.org/articulo.oa?id=32713477006 How to cite Complete issue Scientific Information System More information about this article Network of Scientific Journals from Latin America, the Caribbean, Spain and Portugal Journal's homepage in redalyc.org Non-profit academic project, developed under the open access initiative “main” — 2009/5/4 — 11:00 — page 199 — #1 Anais da Academia Brasileira de Ciências (2009) 81(2): 199-206 (Annals of the Brazilian Academy of Sciences) ISSN 0001-3765 www.scielo.br/aabc Hemoparasites of the genus Trypanosoma (Kinetoplastida: Trypanosomatidae) and hemogregarines in Anurans of the São Paulo and Mato Grosso do Sul States – Brazil DENISE D.M. LEAL1, LUCIA H. O’DWYER1, VITOR C. RIBEIRO2, REINALDO J. SILVA1, VANDA L. FERREIRA3 and ROZANGELA B. RODRIGUES3 1Departamento de Parasitologia, Instituto de Biociências, Unesp, Distrito Rubião Júnior s/n 18618-000 Botucatu, SP, Brasil 2Instituto de Biotecnologia Aplicada a Agricultura, UFV, Campus Universitário, Avenida Peter Henry Rolfs s/n 36570-000 Viçosa, MG, Brasil 3Departamento de Biologia-Ecologia, Centro de Ciências Biológicas e da Saúde (CCBS), UFMS, Cidade Universitária 79070-900 Campo Grande, MS, Brasil Manuscript received on March 11, 2008; accepted for publication on October 8, 2008; presented by LUIZ R. TRAVASSOS ABSTRACT Wild animals are exposed to numerous pathogens, including hemoparasites. The Trypanosoma and hemogregarine group are frequently reported as parasites in anurans (frogs, tree frogs and toads). The identification of these hemopar- asites is usually made through stage observation of their morphology in the peripheral blood of the host. There are no studies, however, based on the biological cycle of these hemoparasites. The objective of the present study was to evaluate the presence of hemogregarines and Trypanosoma spp. in anurans captured in the States of São Paulo and Mato Grosso do Sul – Brazil and to perform the morphological and morphometric characterization of these hemopar- asites. The species of anurans examined were: Dendropsophus nanus, D. minutus, Leptodactylus chaquensis, L. podicipinus, L. labyrinthicus, L. fuscus, Bufo granulosus, B. schneideri, Phyllomedusa hypocondrialis, Trachi- cephalus venulosus, Scinax fuscovarius and Hypsiboas albopunctatus. Of the total of 40 animals studied, four (10%) were positive for hemogregarines and eight (20%) were positive for Trypanosoma spp. Hemogregarine gamonts showed variable morphology and, in addition to intra-erythrocytic forms, extra-erythrocytic forms were also observed. Extremely different forms of Trypanosoma were observed, as described in the literature, with the broad and oval forms being the most common. Key words: Anuran, hemogregarine, Trypanosoma, hemoparasites. INTRODUCTION plexa (Jakes et al. 2003). The main genera of this group The Trypanosoma genus and the hemogregarines have are the Hepatozoon (Family Hepatozoidae) and Haemo- been reported among the hemoparasites infecting anu- gregarina (Family Haemogregarinidae). Differentiation rans. Biological and molecular characterization data between these genera is generally made through the ob- are almost inexistent for these parasites in these animals servation of the sporogonic stage of the parasite in the and species nomination is difficult. The term, hemogre- vector. However, most descriptions have been based garine, is used to describe blood parasites collectively only on the blood stage in the host, while the biologi- belonging to the suborder Adeleorina, Phylum Apicom- cal cycle of the species is not well elucidated. The only completely described species of this genus is Hepata- Correspondence to: Denise Dutra Menezes Leal E-mail: [email protected] zoon catesbianae, a parasite of Rana catesbeiana that, An Acad Bras Cienc (2009) 81 (2) “main” — 2009/5/4 — 11:00 — page 200 — #2 200 DENISE D.M. LEAL et al. in addition to its forms in the vertebrate host, a sporogo- MATERIALS AND METHODS nic development in the Malpighian tubule of the insect Forty amphibian specimens, captured in São Paulo State vector (Desser et al. 1995) has also been described. In (Botucatu, Fartura and Chavantes) and Mato Grosso do Brazil, there is only one report on hemogregarine in the Sul State (Pantanal-Corumbá) were studied. The ani- toad (Bufo crucifer) (Souza and Filho 1974). mals were captured from October to November, 2005 and The genus Trypanosoma belongs to the Kinetoplas- October to November, 2006. The species belong to the tida order and the family Trypanosomatidae. Species family Hylidae (tree frog), family Leptodactylidae (frog) classification of anurans Trypanosoma is confusing, due and family Bufonidae (toad). Animals were collected to the great polymorphism present in this genus and the by hand, blood was taken from their hearts and blood absence of studies on the biological cycle. Since numer- smears were prepared. Smears were fixed in absolute ous species have been described, it is plausible that the methanol for 3 minutes and stained with 10% Giemsa same parasite has received different names in different for 30 minutes. The blood smears were examined with hosts and different geographic localizations. Moreover, an optical microscope, 1,000X magnification, for the exogenous influences, such as temperature, maturity and diagnosis of hemoparasites. The morphometrical analy- density of the population of hosts and environment pol- sis of the parasites was performed using a computerized lution can influence the different shapes of the hemo- system of image analysis (Lite Qwin 3,01 – Leica). parasite (Bardsley and Harmsen 1973, Mackenzie 1999, The variables recorded for hemogregarines were: Silva et al. 2005). Body Area (BA), Body Length (BL) and Body Width The possible vectors of Trypanosoma spp. have (BW); Area (NA), length (NL) and width of the nucleus not yet been identified, but according to previous de- of the parasite (NW). For the Trypanosoma genus, the scriptions, they may include hematophagous dipteran following traits were analyzed: Body Area (BA), Body and leeches (Barrow 1953, Woo 1969, Desser et al. Length excluding the free flagellum (BL) and width of the 1975). Species of Phlebotomus have been shown to parasite (BW); Width at the widest part of body includ- be vectors of this genus in frogs from China (Feng and ing the undulating membrane (W); Area of the nucleus Chao 1943) and in California (Ayala 1971). In Brazil, (NA); Distance from the end anterior to the middle of a recent report has described the infection of this genus the nucleus (A-N); Distance from the posterior end to in anurans. Ferreira et al. (2007) observed the morphol- middle of the nucleus (P-N); Distance from the anterior ogy of the trypanosomes of different biomes in Brazil, end to the kinetoplast (K-A); Distance from the posterior and also genetically studied the genera of 25 isolated end to the kinetoplast (K-P); Distance from the kineto- species of anurans. The authors demonstrated that the plast to the middle of the nucleus (K-N); Distance of the restriction of the parasite to the host is not absolute and kinetoplast to the posterior end of the nucleus (K-Np); showed that some genotypes of Trypanosoma are shared Total length including free flagellum (TF); Length of the by more than one species of different genera and, sporad- undulating membrane (LMO) and Free flagellum (FF). ically, by species from distinct families of anurans. Some The experiments related to this paper have been ap- genotypes were also seen to be restricted to certain local- proved by the Unesp Bioscience Institute’s Ethics Com- ities and were different from those of other trypanosomes mittee approval. The animals were captured according to previously studied in the rest of the world. the IBAMA (Brazilian Institute of Environment and Re- The knowledge of the hemoparasites of anurans is newable Natural Resources) license no. 099/99-DIFAS. of importance, since these animals are in a process of ex- And they were sacrificed with overdose of anaesthetic. tinction and also are extremely sensitive to environmen- tal variations (Silvano and Segalla 2005). The objective RESULTS of this work was to observe the occurrence of hemogre- Of the 40 animals examined, 10% were positive for garines and Trypanosoma spp. in anurans, and to report hemogregarines and 20% were positive for Trypano- the morphological and morphometrical characterization soma spp. (Fig. 1). Among all positive samples one of the hemoparasites found. was chosen as a representative as shown in this paper. An Acad Bras Cienc (2009) 81 (2) “main” — 2009/5/4 — 11:00 — page 201 — #3 TRYPANOSOMA AND HEMOGREGARINES IN ANURANS 201 Positive individuals Number of species Positive individuals Positive individuals for hemogregarines by family for hemogregarines for Trypanosoma and Trypanosoma Leptodactylidae* 2 — 1 (n = 5) Bufonidae* — 1 — (n = 11) Hylidae* 1 — 4 (n = 24) (*) Leptodactylidae: Leptodactylus chaquensis, L. podicipinus, L. labyrinthicus, L. fuscus; Bufonidae:
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