Herpetology Notes, volume 13: 165-168 (2020) (published online on 25 February 2020)

Nematode parasites of hahneli (Boulenger, 1884) from the Eastern Brazilian Amazon

Gabriel Lima Rebêlo¹, Ana Nunes dos Santos¹, Yuri Willkens¹, Lilian Cristina Macedo2, Jeannie Nascimento dos Santos¹, and Francisco Tiago de Vasconcelos Melo1,*

Ameerega hahneli Boulenger, 1884 (Fig.1) Sou et al., 2018). Species of these genera are intestinal is a terrestrial species of the family parasites of anurans, with monoxenic life cycles, and Dendrobatidae Cope, 1865 (1850), and is distributed the transmission routes occur mainly by the active across the Amazon region in the lowlands of , penetration of larvae through the skin (Cosmocerca) or , , , , the extreme south by oral route (Aplectana) (Zaracho et al., 2012). of , southwest and Even though A. hahneli is a well-known Amazonian (Frost, 2019). This species has diurnal habits and species, so far, no studies have been conducted on the occurs in non-flooded forests and “várzea” floodplains, helminth fauna of A. hahneli. The goal of this study is commonly associated with fallen palm tree fronds, to record the occurrence of nematode parasites in A. branches, and small gaps in the forest (Rodrigues and hahneli from the eastern Brazilian Amazon. Duellman, 1994). The studies of A. hahneli, in general, In March 2013, during an expedition to study are related to the comprehension of its behavioural, and reptiles, six specimens of A. hahneli ecological, species population, morphological and from the Reserva de Desenvolvimento Sustentável phylogenetic aspects. Data regarding parasitism in the Amanã, Municipality of Tefé, Amazonas State, Brazil species is still very scarce. (-3.0395ºS, -64.8614ºW), were manually collected, In the groups of parasites commonly found amidst taken to the laboratory, anesthetized and euthanized amphibians, nematodes are the most abundant, being to search for helminth parasites. The helminths found reported in various organs of the digestive tract, the were fixed and stored in 70% ethanol following lungs and blood of different hosts (González et al., established protocols according to Gardner et al. (2012). 2012). Among the main groups of nematode parasites of In the laboratory, nematodes were cleared in Aman amphibians in the Neotropics, the family Cosmocercidae Travassos, 1925 is one of the most diverse, and the genera Aplectana and Cosmocerca are commonly found as parasites of this group of hosts, with 24 and 11 known species, respectively (Campião et al., 2014;

¹ Laboratório de Biologia Celular e Helmintologia “Prof. Dra Reinalda Marisa Lanfredi”, Instituto de Ciências Biológicas, Universidade Federal do Pará, Avenida Augusto Corrêa 01, Guamá, Belém, Pará 66075-110, Brazil. 2 Curso de Ciências Biológicas, Campus Universitário de Capitão Poço, Universidade Federal Rural da Amazônia, Travessa Pau Amarelo, S/n, Vila Nova, Capitão Poço, Pará, 68650-000, Brazil. Figure 1. Lateral view of an adult specimen of Ameerega * Corresponding author. E-mail: [email protected] hahneli Boulenger, 1884. 166 Gabriel Lima Rebêlo et al. lactophenol and examined under an Olympus BX41 Vaucher (1984), fundamental to categorize the species Light Microscope (Olympus, Tokyo, Japan) equipped (Fig. 2). with a drawing tube and an Olympus BX53 with an In the Neotropical region, C. podicipinus has been image capturing system. Voucher specimens were reported in several anuran families, among them: deposited in the invertebrate collection of the Museu Aromobatidae, Bufonidae, Hylidae, Dendrobatidae, Paraense Emílio Goeldi – MPEG. Leptodactylidae, Odontophrynidae and Craugastoridae, Five specimens of A. hahneli were infected with with records in Argentina, Brazil, Colombia, Paraguay nematodes belonging to the family Cosmocercidae and Peru (Campião et al., 2014). Our specimens are (prevalence of 83.3%). In total, eighteen nematode morphologically similar to C. podicipinus parasites specimens were found and attributed to the digestive of Rhinella bergi reported by González and Hamann tract (small and large intestines). One host was found (2007), differing only in the length of the gubernaculum, infected with Cosmocerca podicipinus Baker and which was considered as an intraspecific variation (see Vaucher (1984) (MPEG 00222, Mean Intensity: 1, Table 1). Four species of the genus Cosmocerca are and Mean abundance: 0.16, n = 1). The nematode was reported in Ameerega species: Cosmocerca longispicula identified based on the morphology of a male, regarding (Toledo, 2017) and Cosmocerca parva in A. trivittata presence of five papillae ornamented with fused (Campião et al., 2014); Cosmocerca brasiliense in A. plectanes and rosette tips on the caudal region, which parvula; Cosmocerca parva in A. picta (Campião et al., is, according to the characters described by Baker and 2014) and Cosmocerca sp. in A. pulchripecta from the

1

Table 1. Morphometric data of Cosmocercid nematodes of Ameerega hahneli from the Brazilian Amazon (all measurements are given in micrometres,Table 1. Morphometric except when data indicated). of Cosmocercid nematodes of Ameerega hahneli from the Brazilian Amazon (all measurements are given in micrometres, except when indicated).

Cosmocerca podicipinus Aplectana elenae Cosmocercids females (n = 1) (n = 2) (n = 6) Total length 2.56 mm 1.7–2 mm 2.12 mm (2–2.3) Oesophagus length 184 341–406 403 (386.8–417.4) Pharynx length 23 25–30 35 (28.5–40.2) Muscular esophagus 175 251–298 286 (268.7–295.7) Bulb length 61 65–78 82 (77.9–87) Bulb width 48 70–102 96 (83.1–102.5) Nerve ring from anterior end 133 143–166 166 (150.6–183.1) Excretory pore from anterior end 227 200–277 229 (214.2–238.9) Tail length 161 235–276 238 (187–253.2) Gubernaculum 65 30–55 - Spicules 135 87–98 - Eggs length - - 55.87 (51.6–61.9) Eggs width - - 36.98 (35.3–38.8) Vulva from anterior end - - 1.05 mm (0.9–1.1) Papilla arrangement (pre:ad:post 5 + 1:3:5 16 + 1:2:10 - anal ) Nematode parasites of Ameerega hahneli from the Eastern Brazilian Amazon 167

Figure 2. Cosmocerca podicipinus Baker and Vaucher, 1984 parasite of A. hahneli from the Brazilian Amazon (male). a – Anterior extremity of the body (50 µm); b – View of the anterior third of the esophagus, with emphasis on the pharynx region (20 µm); c – Posterior extremity, with emphasis on precloacal papillae (arrows indicate precloacal papillae) (50 µm); d – Detail of papillae ornamented with fused plectanes and rosette tips (40 µm); e – Detail of spicules and gubernaculum (40 µm). 168 Gabriel Lima Rebêlo et al.

Brazilian Amazon (Tavares-Costa et al., 2019). References

One host was found with Aplectana elenae Baker and Baker, M.R., Vaucher, C. (1984): Parasitic helminths from Paraguay Vaucher (1986) infecting the small intestines (MPEG VI: Cosmocerca Diesing, 1861 (Nematoda: Cosmocercoidea) 00221, Mean Intensity: 2; and Mean abundance: 0.33). from . Revue Suisse de Zoologie 91: 925–934. The presence of simple caudal papillae without rosettes Baker, M.R., Vaucher, C. (1986): Parasitic helminths from or plectanes, their number and distribution, the length Paraguay XII: Aplectana Railliet & Henry, 1916 (Nematoda: of the spicule and gubernaculum were considered Cosmocercoidea) from frogs. Revue Suisse de Zoologie 93: 607–616. to identify this species, as proposed by Gomez et al. Campião, K.M., Morais, D.H., Dias, O.T., Aguiar, A., Toledo (2017). G.M., Tavares, L.E.R., Silva, J.R. (2014): Checklist of helminth In addition, our morphological and morphometric data parasites of amphibians from South America. 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Zootaxa 4472: 194– of the ecology and biology of the species of Ameerega 200. from the Brazilian Amazon, considering that out of 31 Tavares-Costa, L.F.S., Dias-Souza, M.R., Costa-Campos, C.E., known species (Frost, 2019), only A. parvula, A. picta, Melo F.T.V (2019): Helminth parasites of Ameerega pulchipecta A. trivittata and A. pulchripecta were examined for (Anura: Dendrobatidae) from the eastern Amazon, Brazil. parasites. Herpetology Notes 12: 435–436. Toledo, G.M. (2017): Biodiversity of parasitic helminths of anurans Acknowledgments. We thank T.C.S. de Ávila Pires of the Emílio from different phytophysiognomies in Peru. Unpublished PhD Goeldi Museum for the invitation to L.C. Macedo to take part in thesis. Universidade Estadual Paulista “Júlio De Mesquita field work, and Leandra Cardoso Leal and all the herpetofauna Filho”, Botucatu, Brasil. group. We appreciate the help of Cybelle Albuquerque de Miranda Zaracho, V.H., Acosta, L.J., Lamas, F. (2012): Dieta y parasitismo on the revision of the manuscript. We are also grateful to Wallace de Leptodactylus diptyx (Anura: Leptodactylidae) from Douglas Garcia das Neves for the photograph of Ameerega Northeastern Argentina. Revista Mexicana de Biodiversidad 83: hahneli. This study was financially supported by the Conselho 1180–1186. Nacional de Desenvolvimento Cientifico e Tecnológico – CNPq (grant n. 431809/2018-6 Universal); J.N. Santos and F.T.V. Accepted by Fabrício Oda Melo would like to thank CNPq for having provided a research fellowship (process n. 304955/2018-3).