Herpetology Notes, volume 11: 367-368 (2018) (published online on 02 May 2018)

First record of Filarial nematode in the Brazilian torrent frog Hylodes heyeri (Anura, Hylodidae)

Elvira D’Bastiani1,*, Michelle M. Struett1, Maurício O. Moura1 and Karla M. Campião1

Anura is the most diverse amphibian order, with a of the Museu de História Natural do Capão da Imbuia recognized importance in biodiversity and conservation. (MHNCI 10868-10872) in Curitiba, Paraná, Brazil. However, we still lack information about its interaction We examined the blood smears of twenty specimens with different trophic levels, such as parasitism. of H. heyeri (mean SVL = 40.91 mm ± 0.15 SD, mean Although we have advanced significantly in the last body weight = 5.75 g ± 0.75 SD). Among those, one years, we have studied host-parasite interactions for only specimen (male, SVL = 40.29 mm, weight = 6.9 g) 16.45% of the Brazilian anurans (Campião et al., 2014). was infected by microfilariae (n = 13, average length Among helminth endoparasites, the filaroid nematodes = 100.14 µm, ± 25.36 SD, average width = 4.91 µm are a very peculiar group, once they include a life stage ± 1.19 SD; Fig. 2). in the blood of their hosts. Blood parasites also include Infections by blood nematode parasites are reported protozoans such as Trypanosoma, Hepatozoon and among amphibians, reptiles, birds and mammals Haemogregarina species (Souza, 1974; McKenzie and (McKenzie, 2007). In the most common form of Starks, 2008). infection, parasites in adult stage live within the The Hylodidae Günther, 1858 represents well the host’s body cavity, where the female releases the scarcity of studies, since it is composed by 26 species larval microfilariae that reach the bloodstream of the occurring in the southeast and south of Brazil (Frost, host (McKenzie, 2007). Among amphibians, reports 2017) and none of these was studied for parasites. of microfilariae are scarce. Only Rhinella marina Here we report the association of a filarial nematode (Linnaeus, 1758), Boana lundii (Burmeister, 1856), parasitizing Hylodes heyeri (Fig. 1). Our study was Boana geographica (Spix, 1824), Bokermannohyla carried out in Parque Estadual do Marumbi (MSP, luctuosa (Pombal and Haddad, 1993), and Leptodactylus State Decree 7.300/90) (25°29’23” S; 48°58’37” W) in Piraquara, Paraná, southern Brazil. We conducted two field surveys during the breeding season, one in September 2017 and one in January 2018. We surveyed two streams using visual and auditory surveys (Scott and Woodward, 1994), license SISBIO 53210-2. For each anuran we collected blood samples and made blood smears, which were fixed with methanol, stained with Giemsa (16%), and screened for blood parasites with an optical microscope (Carl Zeiss). Then, specimens were killed with Lidocaine 5%, fixed in 10% formalin, preserved in 70% ethanol and deposited in the collection

1 Programa de Pós-Graduação em Ecologia e Conservação, Universidade Federal do Paraná, CEP 81531-980, Curitiba, Figure 1. Adult male of Hylodes heyeri from Parque Estadual Paraná, Brazil. do Marumbi, in the municipality of Piraquara, State of Paraná, * Corresponding author. E-mail: [email protected] southern Brazil. Photo by Michelle M. Struett. 368 Elvira D’Bastiani et al.

parasites of Amphibians from South America. Zootaxa 3843: 1-93. Causey, O.R. (1939): Aedes and Culex mosquitoes as intermediate hosts of frog filaria, Foleyella sp. American Journal of Hygiene 29: 2. Frost, D.R. (2017): Amphibian species of the world: an online reference. Versão 6.0 (Accessed October 25, 2017). American Museum of Natural History, New York, EUA. Available at: http://research.amnh.org/vz/herpetology/amphibia/Amphibia/ Anura/Hylodidae/Hylodes/Hylodes-heyeri. Accessed 27 February of 2018. Haddad, C.F., Pombal Jr, J.P., Bastos, R.P. (1996): New species of Hylodes from the Atlantic forest of Brazil (Amphibia: Leptodactylidae). Copeia 4: 965-969. McKenzie, V.J. (2007): Human land use and patterns of parasitism in tropical amphibian hosts. Biological Conservation 137: 102- Figure 2. Filarial parasite of Hylodes heyeri from Parque 116. Estadual do Marumbi, State of Paraná, southern Brazil. Photo McKenzie, V.J., Starks, H.A. (2008): Blood parasites of two Costa by Elvira D’Bastiani. Rican amphibians with comments on detection and microfilaria density associated with adult filarial worm intensity. The Journal of Parasitology 94: 824-829. Scott JR., N.J., Woodward, B.D. (1994): Surveys at breeding sites. latrans (Steffen, 1815) have been reported as hosts of In: Measuring and monitoring biological diversity: Standard microfilariae in Brazil (Travassos and Freitas, 1964; methods for amphibians, p. 84-92. Heyer, W.R., Donnelly, Vicente and Santos, 1976; Vicente et al., 1991; Toledo M.A., McDiarmid, R.W., Hayek, L.A.C., Foster, M.S. Eds., Washington, Smithsonian Institution Press. et al., 2013). Therefore, this is the first record of Souza, M.A., Borriello Filho., A. (1974): Uma nova hemogregarina microfilariae larvae in a hylodid species. no sangue de Bufo crucifer WIED, 1821 do Brasil. Mem Inst Hylodes heyeri is a rheophilic species (Haddad et al., Oswaldo Cruz 72: 275-282. 1996), and the aquatic environment that it inhabits may Toledo, G.M., Aguiar, A., Silva, R.J., Anjos, L.A. (2013): Helminth contribute to infections by hemoparasites. Nematodes of Fauna of Two Species of Physalaemus (Anura: Leiuperidae) the Onchocercidae family are the main filarid parasites from an Undisturbed Fragment of the Atlantic Rainforest, associated with amphibians, and are usually transmitted Southeastern Brazil. Journal of Parasitology 99: 919-922. Travassos, L., Freitas, J.F.T. (1964): Pesquisas helmintológicas by hematophagous arthropods (Anderson, 2000). realizadas em Maicujú, Estado do Pará. Publicações Avulsas do Species of Culex and Aedes (Culicidae) are vectors Museu Paraense Emílio Goeldi 1: 3-16. of microfilariae larvae infecting frogs (Causey, 1939; Vicente, J.J., Santos, E. (1976): Fauna helmintológica de McKenzie, 2007). It is possible that Culex and Aedes Leptodactylus ocellatus de Volta Redonda, Estado do Rio de are the main vectors of microfilariae for H. heyeri in Janeiro. Atas da Sociedade de Biologia do Rio de Janeiro 18: our study area. Further studies should investigate how 27-42. the microfilariae can affect H. heyeri, and how this Vicente J.J., Rodrigues, H.O., Gomes, D.C., Pinto, R.M. (1991): Nematóides do Brasil 2a hemoparasite is transmitted. parte: Nematóides de Anfíbios. Revista Brasileira de Zoologia 7: 549-626.

Acknowledgments. We thank Parque Estadual do Marumbi for the facilities and sampling license (No. 41.16); Dr. Mário Navarro for providing the microscope for the picture; Gabriel de La Torre for helping to stain the slides; André Bruinje, Fabrício H. Oda and Drausio H. Morais for reviewing this manuscript.

References

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