A New Metastrongyloidean Species (Nematoda) Parasitizing Pulmonary Arteries of Puma (Herpailurus) yagouaroundi (É. Geoffroy, 1803) (Carnivora: Felidae) from Brazil Author(s): Fabiano M. Vieira , Luís C. Muniz-Pereira , Sueli de Souza Lima , Antonio H. A. Moraes Neto , Erick V. Guimarães , and José L. Luque Source: Journal of Parasitology, 99(2):327-331. 2013. Published By: American Society of Parasitologists DOI: http://dx.doi.org/10.1645/GE-3171.1 URL: http://www.bioone.org/doi/full/10.1645/GE-3171.1
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J. Parasitol., 99(2), 2013, pp. 327–331 Ó American Society of Parasitologists 2013
A NEW METASTRONGYLOIDEAN SPECIES (NEMATODA) PARASITIZING PULMONARY ´ GEOFFROY, 1803) ARTERIES OF PUMA (HERPAILURUS) YAGOUAROUNDI (E. (CARNIVORA: FELIDAE) FROM BRAZIL Fabiano M. Vieira, Lu´ıs C. Muniz-Pereira*, Sueli de Souza Lima†, Antonio H. A. Moraes Neto‡, Erick V. Guimara˜es§, and Jose´ L. Luquejj Curso de P´ os-Gradua¸ca˜o em Biologia Animal, Universidade Federal Rural do Rio de Janeiro, Serop´edica, RJ, CEP: 23890-000, Brazil. Correspondence should be sent to:
[email protected] ´ ABSTRACT: Angiostrongylus felineus n. sp. (Nematoda, Metastrongyloidea), parasitic in Puma (Herpailurus) yagouaroundi (E. Geoffroy, 1803) (Carnivora, Felidae) from the municipality of Juiz de Fora, Minas Gerais state, Brazil, is described and illustrated herein. Angiostrongylus felineus n. sp. differs from all congeneric species by having the anterior extremity with accentuated cuticular expansion and by smaller size of spicules. This study describes for the first time a species of Angiostrongylus in a wild Felidae in Brazil. The parasites were fixed AFA for 48 hr and preserved in 70% ethanol with 5% glycerin. For light microscopy studies, the nematodes were cleared in Amann’s lactophenol, in which they were maintained while measurements were taken. Drawings were made with the aid of a drawing tube attached to the Olympus BX41 light microscope (Olympus America Inc., Melville, New York). Measurements are given as ranges in micrometers, with the mean 6 standard deviation in parentheses. Photomicrographs were made with a Canon A3000IS camera (Canon Singapore Pte. Ltd., Keppel Bay Tower, Singapore), with 10 megapixels of resolution, coupled to the Olympus BX41 light microscope. Some photomicrographs were made using a compound Olympus BX51 light microscope (Olympus America Inc.) equipped with Nomarski differential interference contrast (DIC) optics. For scanning electron microscopy (SEM) the nematodes were fixed in a solution containing 2.5% glutaraldehyde, 4% paraformaldehyde, and 5 mM calcium chloride in 0.1 M cacodylate buffer (pH 7.2) at room temperature, then washed with cacodylate buffer, postfixed in 1% OsO4 in 0.1 M cacodylate buffer containing 0.8% potassium ferricyanide and 5 mM CaCl2, rinsed again with cacodylate buffer, dehydrated in ethanol, processed in a critical-point drier with CO2, sputter-coated with gold, and examined in a JEOL JSM 6390LV SEM (JEOL Ltd., Tokyo, Japan) operating at 15 kV. Identification and classification of the specimens to the generic level follow Chabaud (1974), Anderson (1978), Ubelaker (1986), and Gibbons (2010). The holotype, allotype, and paratypes are deposited in the Instituto Oswaldo Cruz Helminthological Collection (CHIOC), Rio de Janeiro, Brazil.
Species of Angiostrongylus Kamensky, 1905 parasitize rodents, shrews, and carnivores (Anderson, 2000). In Brazil, species in this genus have been reported in wild and domestic mammals of the orders Carnivora and Rodentia (Vicente et al., 1997; Vieira et al., 2008). There are no previous records of Angiostrongylus spp. in mammals of the family Felidae (Carnivora) from Brazil. ´ Geoffroy, 1803) (CarPuma (Herpailurus) yagouaroundi (E. nivora, Felidae) is a medium-sized feline found from Texas in the United States throughout most of South America except Uruguay and Chile (Ximenez, 1972; Eisenberg, 1989; Oliveira, 1998). In Brazil this species is found in all biomes (Oliveira, 1998). It is predominantly diurnal and has a typical diet of a carnivorous specialist, feeding on reptiles, birds, lagomorphs, rodents, other small mammals, and carrion (Oliveira, 1998). The only species of Metastrongyloidea reported in this host in Brazil is Aelurostrongylus abstrusus (Railliet, 1898) (Angiostrongylidae) (Vicente et al., 1997; Noronha et al., 2002; Vieira et al. 2008). The aim of the present study is to describe a new species of Angiostrongylus in wild felids in Brazil. MATERIALS AND METHODS The nematodes were collected from pulmonary arteries during necropsy of P. (H.) yagouaroundi from the municipality of Juiz de Fora (21840 0 22.03 00 S, 43826 0 39.07 00 W), Minas Gerais State, Brazil. The hosts examined were all ‘‘road kills’’ and were donated by the Regional Office of the Instituto Brasileiro do Meio Ambiente e dos Recursos Naturais Renova´veis (IBAMA), in the municipality of Juiz de Fora, MG, in 2002. The hosts were identified according to Oliveira (1998) and Cheida et al. (2006).
DESCRIPTION Angiostrongylus felineus n. sp. (Figs. 1–13) General: Body of both sexes filiform, with pronounced sexual dimorphism; females larger than males. In both sexes, mouth aperture not surrounded by lips, and evident papillae (Figs. 12, 13). Anterior extremity with accentuated cuticular expansion (Figs. 1, 4, 6, 10–13). Esophagus claviform (Fig. 1). Excretory pore slightly posterior to end of esophagus. Nerve ring in middle region of esophagus (Fig. 1). Male (holotype and 4 paratypes): Total length 16.15–18.07 (16.91 6 0.8) mm. Width at level of base of the esophagus 285–355 (321 6 25). Esophagus 275–315 (296 6 17) long. Anterior end of testis variable in position. Caudal bursa symmetrical and poorly developed. Ventral rays equal, fused in lower three-fourths. Ventral stem separated from lateral stem (Figs. 2, 7). Anterolateral rays digitiform, separated from other 2 laterals (Figs. 2, 5, 7). Mediolateral and posterolateral rays with common stem, and separate in distal half (Figs. 2, 5, 7). Externodorsal ray digitiform and slightly longer than dorsal ray (Figs. 2, 5, 7). Dorsal ray with 2 equal branches, separate in distal half (Figs. 2, 7). Spicules elongated, equal, similar 360–400 (383 6 21). Distal ends of spicules sharply pointed. Gubernaculum inconspicuous, a little chitinized, and curved. Ratio of spicule length:body length 46:1. Female (allotype and 3 paratypes): Body length 20–24 (23 6 2) mm. Width at base of esophagus 300–360 (326 6 26). Esophagus 260–390 (333 6 56) long. Anterior ends of ovaries variable in position, sometimes reaching to esophagus–intestinal junction (Fig. 1). Uterine tubules spirally
Received 10 April 2012; revised 17 September 2012; accepted 27 September 2012. ´ * Laboratorio de Helmintos Parasitos de Vertebrados, Instituto Oswaldo Cruz, FIOCRUZ, Rio de Janeiro, RJ, CEP 21040-360, Brazil. ´ † Departamento de Zoologia, Laboratorio de Taxonomia e Ecologia de Helmintos, Universidade Federal de Juiz de Fora, Juiz de Fora, MG, CEP 36036-900, Brazil. ´ ‡ Laboratorio de Inovac¸ oes em Terapias, Ensino e Bioprodutos, ˜ Instituto Oswaldo Cruz, FIOCRUZ, Rio de Janeiro, RJ, CEP 21040-360, Brazil. ´ § Laboratorio de Biologia Estrutural, Instituto Oswaldo Cruz, FIOCRUZ, Rio de Janeiro, RJ, CEP 21040-360, Brazil. jj Departamento de Parasitologia Animal, Universidade Federal Rural ´ do Rio de Janeiro, Caixa Postal 74.508, CEP 23851-970, Seropedica, RJ, Brazil. DOI: 10.1645/GE-3171.1 327
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FIGURES 1–5. Angiostrongylus felineus n. sp. (1) Female, anterior region, ventral view. (2) Male, caudal bursa, ventral view. (3) Female, posterior region, lateral view. (4) Male, anterior region, lateral view. (5) Male, caudal bursa, ventral view. Detail of the lateral ray–anterolateral ray independent of the other 2 lateral rays.
VIEIRA ET AL.—ANGIOSTRONGYLUS N. SP. FROM PUMA (HERPAILURUS) YAGOUAROUNDI
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FIGURES 6–9. Light microscopy of Angiostrongylus felineus n. sp. (6) Male, anterior region, lateral view. (7) Male, caudal bursa, ventral view. (8) Female, posterior region, lateral view. (9) Female, posterior region, lateral view (DIC microscopy). wound around intestine, producing ‘‘barber-pole’’ appearance typical of genus. Vulva with slightly elevated lips (Figs. 3, 8, 9), 280–325 (296 6 20) from tip of tail. Tail 85–110 (96 6 13) long. Tail bluntly rounded (Figs. 3, 8, 9). Eggs not observed at level of ovojector. Taxonomic summary ´ Geoffroy, 1803) Type host: Puma (Herpailurus) yagouaroundi (E. (Carnivora: Felidae) (jaguarundi). Site of infection: Pulmonary arteries. Intensity: Twenty-nine, all adult. Type locality: Juiz de Fora, Minas Gerais, Brazil, 21841 0 20 00 S, 43820 0 40 00 W. Type specimens: Holotype male CHIOC no. 35812a; allotype female CHIOC no. 35812b; and 6 paratypes CHIOC no. 35812c (male) and 35812d (female) (preserved in 70% ethanol with 5% glycerin). Etymology: The specific name was chosen because this species was the first Angiostrongylus described and reported in a wild Felidae from Brazil. Remarks According to Costa et al. (2003), Robles et al. (2008), and Souza et al. (2009), who adopted the criteria of Drozdz (1970) and Anderson (1978) for the taxonomy of Angiostrongylus, this genus has 17 species that are differentiated principally by the morphology of the rays of the copulatory bursa of males and by morphometric characters. Angiostrongylus felineus n. sp. possesses the anterolateral ray of the copulatory bursa originating independently from the common stem of the mediolateral and posterolateral rays. This species differs from Angiostrongylus gubernaculatus Dougherty, 1946; Angiostrongylus taterone
Baylis, 1928; Angiostrongylus cantonensis Chen, 1935; Angiostrongylus sciuri Merdevenci, 1964; Angiostrongylus mackerrasae Bhaibulaya, 1968; Angiostrongylus sandarsae Alicata, 1968; Angiostrongylus petrowi Tarjymanova and Tschertkova, 1969; Angiostrongylus dujardini Drozdz and Doby, 1970; Angiostrongylus schmidti Kinsella, 1971; Angiostrongylus ´ costaricensis Morera and Cespedes, 1971; Angiostrongylus malaysiensis Bhaibulay and Cross, 1971; Angiostrongylus ryjikovi Jushkov, 1971; Angiostrongylus siamensis Ohbayashi, Kamiya, and Bhaibulaya, 1979; Angiostrongylus morerai Robles, Navone, and Kinsella, 2008, and Angiostrongylus lenzii Souza, Simoes, Thiengo, Lima, Mota, Rodrigues˜ Silva, Lanfredi, and Maldonado, 2009 by those species possessing the 3 lateral rays originating from the common stem (Robles et al., 2008; Souza et al., 2009). Along with Angiostrongylus felineus n. sp., another 2 species of this genus have the anterolateral ray originating independently from the common stem of the mediolateral and posterolateral rays: Angiostrongylus vasorum (Baillet, 1866) and Angiostrongylus chabaudi Biocca, 1957. Angiostrongylus felineus n. sp. differs from these other 2 species of the genus in having an accentuated cuticular expansion in the cephalic extremity (Figs. 1, 4, 6, 10–15). This characteristic was not observed in these other 2 species of Angiostrongylus. Moreover, a morphometric characteristic also demonstrates the differences between species, i.e., A. vasorum and A. chabaudi have larger spicules (Biocca, 1957; Rosen et al., 1970; Guilhon and Cens, 1973) in comparison to those in A. felineus n. sp. (Table I). This is most evident when comparing the proportional size of the spicules in relation to the total body length (spicule length:body length ratio) in these 3 Angiostrongylus species (Table I).
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FIGURES 10–11. DIC microscopy of Angiostrongylus felineus n. sp. (10) Male, anterior region, ventral view. (11) Female, anterior region, lateral view.
DISCUSSION The systematics of Angiostrongylus and their synonymy needs a revision because, for some authors (i.e., Robles et al., 2008; Souza et al., 2009), the morphology of the lateral rays of the copulatory bursa of males does not justify the differentiation between genera, whereas for others (i.e., Ubelaker, 1986; Gibbons, 2010) such characters are valid. The genus Angiostrongylus Kamensky (1905) (Metastrongyloidea, Angiostrongylidae) was proposed to accommodate Strongylus vasorum described by Baillet (1866). The diagnostic character of this genus is the morphology of the lateral rays of the copulatory bursa, with the antero-lateral ray separate from the medio-lateral and postero-lateral rays, which share a common stem and separate in the distal half (Kamensky, 1905; Ubelaker, 1986; Gibbons, 2010). Ubelaker (1986) considers this character enough to differentiate the genera Angiostrongylus, Parastrongylus Baylis, 1929 (Metastrongyloidea, Angiostrongylidae), and Angiocaulus Schulz, 1951 (Metastrongyloidea, Angiostrongylidae), which Drozdz (1970) and Anderson (1978) considered as a synonym of Angiostrongylus. The most recent study by Gibbons (2010) considers the proposal suggested by Ubelaker (1986) to be valid, restoring the validity of Parastrongylus, which Anderson (1978) considered a subspecies of Angiostrongylus and Angiocaulus. However, recent studies of Costa et al. (2003), Robles et al. (2008), and Souza et al. (2009) did not consider that the proposal for Ubelaker (1986) was valid because some specimens they studied of A. vasorum, the type species of the genus, exhibited variable morphology of the lateral rays of the copulatory bursa. The anterolateral ray may, or may not, be in the same common stem of the mediolateral and posterolateral rays (Robles et al., 2008; Souza et al., 2009). It is worth pointing out the validity or not of a Brazilian species of Angiostrongylus. According to Grisi (1971), Angiostrongylus
raillieti (Travassos, 1927) (¼Angiocaulus raillieti [Travassos, 1927]), described by Travassos (1927) as Haemostrongylus raillieti, a parasite of the pulmonary arteries of Cerdocyon thous (Linnaeus, 1766) (Carnivora, Canidae) in Brazil, should be considered a valid species and not a synonym of A. vasorum from Brazil. Grisi (1971) discusses the hypothesis that A. vasorum is a species that does not occur in Brazil and is actually A. raillieti. The molecular study conducted by Jefferies et al. (2009) supports the idea that A. vasorum does not occur in Brazil. Based on the comparison of molecular characters of specimens of A. vasorum from Europe and South America, Jefferies et al. (2009) demonstrated that genetic variations in the nematodes from the 2 continents indicate different lineages and are not the same species. According to we authors, the proposal suggested by Costa et al. (2003) considering A. raillieti a synonym of A. vasorum was premature because the morphological similarities observed by Costa et al. (2003) between A. vasorum from Europe (studied by Rosen et al. [1970] and Guilhon and Cens [1973]) and the supposed A. vasorum from Brazil can be justified as an event of cryptic speciation; and that the Brazilian species is likely A. raillieti. Until the present study, a few species of Metastrongyloidea have been reported in wild carnivorous mammals in Brazil (Vicente et al., 1997; Vieira et al., 2008). Angiostrongylidae is the only family that has records in wild carnivores in Brazil, with Angiostrongylus raillieti (Travassos, 1927) (¼Haemostrongylus raillieti Travassos, 1927; Angiocaulus raillieti, Angiostrongylus vasorum [Raillet, 1866] sensu Costa et al. [2003]) occurring in C. thous, Lycalopex vetulus (Lund, 1842) (Carnivora, Canidae), and Nasua nasua (Linnaeus, 1766) (Carnivora, Canidae) (Travassos, 1927; Grisi, 1971; Lima et al., 1994; Duarte et al., 2007; Vicente et al., 1997; Vieira et al., 2008) and A. abstrusus occurring in Puma (Herpailurus) yagouaroundi (Noronha et al., 2002; Vieira et al., 2008). Therefore, the present study describes a species of Angiostrongylus for the first time in a wild felid from Brazil.
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FIGURES 12–13. SEM of Angiostrongylus felineus n. sp. (12) Male, anterior region. (13) Male, anterior region. White arrow indicates the buccal opening.
ACKNOWLEDGMENTS Fabiano M. Vieira was supported by a doctoral fellowship from REUNI/UFRRJ (Programa de Apoio ao Plano de Reestruturaca ¸ ˜o e Expansa˜o das Universidades Federais, Brazil); Jose´ L. Luque was supported by a research fellowship from CNPq (Conselho Nacional de ´ Pesquisa e Desenvolvimento Tecnologico, Brazil).
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