image/svg+xml247Helminths Infecting the Carvalho’s EscuerzoNeotropical Helminthology (Lima). Vol. 17, Nº2, jul - dic 2023Neotropical HelminthologyNeotropical Helminthology, 2023, vol. 17 (2), 247-258ORIGINAL ARTICLE / ARTÍCULO ORIGINALHELMINTHS INFECTING THE CARVALHO’S ESCUERZO ODONTOPHRYNUS CARVALHOI FROM THE BRAZILIAN STATE OF CEARÁHELMINTOS QUE INFECTAN EL ESCUERZO DE CARVALHO ODONTOPHRYNUS CARVALHOIDEL ESTADO BRASILEÑO DE CEARÁHELMINTOS INFECTANDO O SAPINHO CARVALHO ODONTOPHRYNUS CARVALHOINO ESTADO BRASILEIRO DO CEARÁTatiana Feitosa Quirino1,*; Dalilange Batista-Oliveira 2; Matheus Calixto Saldanha3& Robson Waldemar Ávila ¹,2ISSN Versión Impresa 2218-6425 ISSN Versión Electrónica 1995-1403DOI: https://dx.doi.org/10.24039/rnh20231721685Volume 17, Number 2 (jul - dec) 2023Este artículo es publicado por la revista Neotropical Helminthology de la Facultad de Ciencias Naturales y Matemática, Universidad Nacional Federico Villarreal, Lima, Perú auspiciado por la Asociación Peruana de Helmintología e Invertebrados Af nes (APHIA). Este es un artículo de acceso abierto, distribuido bajo los términos de la licencia Creative Commons Atribución 4.0 Internacional (CC BY 4.0) [https:// creativecommons.org/licenses/by/4.0/deed.es] que permite el uso, distribución y reproducción en cualquier medio, siempre que la obra original sea debidamente citada de su fuente original.1 Graduate Course in Systematics, Use, and Conservation of Biodiversity, Departament of Biology, Pici Campus, Federal University of Ceará, Fortaleza-CE Zip Code 60440-900, Brazil.2 Graduate Course of Ecology and Natural Resources, Department of Biology, Pici Campus, Federal University of Ceará, Fortaleza - CE Zip Code 60440-900, Brazil.3 Graduate Course Biology, Departament of Biology, Pici Campus, Federal University of Ceará, Fortaleza-CE Zip Code 60440-900, Brazil.* Corresponding author: tata_tatifeitosa@hotmail.comTatiana Feitosa Quirino:https://orcid.org/0000-0001-8184-8705 Dalilange Batista-Oliveira: https://orcid.org/0000-0002-4140-6643Matheus Calixto Saldanha: https://orcid.org/0009-0005-8884-7015Robson Waldemar-Ávila: https://orcid.org/0000-0003-3641-8321ABSTRACTT e family Odontophrynidae comprises 40 anuran species widely distributed in South America, ranging from Brazil to Argentina and Paraguay. OdontophrynuscarvalhoiSavage and Cei, 1965, a medium-sized species with terrestrial habits and explosive reproduction, is one of the representatives of this family. Despite its extensive distribution, data regarding its natural history are limited, with published information primarily focused on activity patterns, diet, and defensive behavior. In this study, we present data on the composition and infection patterns of endoparasites associated with O. carvalhoiin a relictual forest in the Brazilian state of Ceará. T e parasite community comprised 11 species, with Aplectana hylambatis(76.67%) and Oswaldocruzia mazzai(79.49%) being the most prevalent. Additionally, we report new occurrences of parasite species for O. carvalhoi, contributing to the understanding of the parasitic fauna
image/svg+xml248Neotropical Helminthology (Lima). Vol. 17, Nº2, jul - dic 2023Feitosa Quirino et al.in the Neotropical region. Tese fndings underscore the importance of parasitological research for a comprehensive understanding of the ecology and biology of this amphibian species.Keywords: Parasites – Amphibian – Odontophrynidae – Relictual forestRESUMENLa familia Odontophrynidae comprende 40 especies de anuros ampliamente distribuidas en América del Sur, desde Brasil hasta Argentina y Paraguay. Odontophrynus carvalhoiSavage & Cei, 1965, especie de tamaño mediano, hábitos terrestres y reproducción explosiva, es uno de los representantes de esta familia. A pesar de su extensa distribución, los datos sobre su historia natural son limitados, y la información publicada se centra principalmente en patrones de actividad, dieta y comportamiento defensivo. En este estudio, presentamos datos sobre la composición y patrones de infección de endoparásitos asociados con O. carvalhoien un bosque relicto en el estado brasileño de Ceará. La comunidad de parásitos estuvo compuesta por 11 especies, siendo Aplectana hylambatis(76,67%) y Oswaldocruzia mazzai(79,49%) las más prevalentes. Además, reportamos nuevas ocurrencias de especies de parásitos para O. carvalhoi,contribuyendo al conocimiento de la fauna parasitaria en la región Neotropical. Estos hallazgos subrayan la importancia de la investigación parasitológica para una comprensión integral de la ecología y biología de esta especie de anfbio.Palabras clave: Parásitos – Anfbios – Odontophrynidae – Bosque relictoRESUMOA família Odontophrynidae compreende 40 espécies de anfíbios amplamente distribuídas na América do Sul, abrangendo desde o Brasil até a Argentina e o Paraguai. Odontophrynus carvalhoiSavage e Cei, 1965, uma espécie de porte médio com hábitos terrestres e reprodução explosiva, é uma das representantes dessa família. Apesar de sua distribuição extensiva, dados sobre sua história natural são limitados, com informações publicadas concentradas principalmente em padrões de atividade, dieta e comportamento defensivo. Neste estudo, apresentamos dados sobre a composição e padrões de infecção de endoparasitas associados a O. carvalhoiem uma foresta relictual no estado brasileiro do Ceará. A comunidade de parasitas incluiu 11 espécies, sendo Aplectanahylambatis(76,67%) e Oswaldocruziamazzai(79,49%) as mais prevalentes. Adicionalmente, relatamos novas ocorrências de espécies de parasitas para O. carvalhoi, contribuindo para a compreensão da fauna parasitária na região Neotropical. Essas descobertas destacam a importância da pesquisa parasitológica para uma compreensão abrangente da ecologia e biologia dessa espécie de anfíbio.Palavras-chave: Helmintos – Anfíbio – Odontophrynidae – Floresta relictual.
image/svg+xml249Helminths Infecting the Carvalho’s EscuerzoNeotropical Helminthology (Lima). Vol. 17, Nº2, jul - dic 2023INTRODUCTIONTe family Odontophrynidae is composed by 40 anuran species widely distributed in South America, from Brazil to Argentina and Paraguay (Segalla et al., 2019; Mângia et al., 2020; Frost, 2021). Te genus OdontophrynusReinhardt & Lütken, 1862, currently have 12 species (Frost et al., 2017) belonging to three phenetic groups (Savage & Cei, 1965; Caramaschi, 1996; Caramaschi & Napoli, 2012): O. americanus, O. cultripes and O. occidentalis. Besides, O. salvatoriactually is not allocated to any group (Amaro et al.,2009).Odontophrynus carvalhoi, belongs to O. cultripes group, was described by Savage & Cei, 1965, from a single specimen collected in Poção municipality, state of Pernambuco, Brazil. Its distribution encompasses the Jequitinhonha River valley, in Minas Gerais state, to northeastern Goiás state and Ceará state in the north, in phytophisiognomies of Atlantic Forest, Cerrado and Caatinga Biomes (Juncá, 2006; Lisboa et al., 2010; Caramaschi & Napoli, 2012; Dias et al., 2014; Santos et al.,2017). It is a medium sized species, with terres-trial habits and explosive reproduction(Lynch, 1971; Freitas & Silva, 2004; Caramaschi & Napoli, 2012). Tadpoles are laid in small streams inside forested habitats (Santos et al., 2017).Despite its huge distribution, data on natural history of O. carvalhoi are scant, with published information about activity patterns and diet (Brito et al., 2012) and defensive behavior (Bezerra et al., 2010; Borges-No-josa et al., 2016). Until now, there is no information regarding endoparasitesinfecting O. carvalhoi, and several studies pointed out the importance of knowledge of parasites given the role of these organisms in ecosystem regulation. In particular, the diversity of helminths associated with amphibians is considered rich and diversifed, although it is a hidden diversity and therefore undersampled (Poulin, 2014; Campião et al., 2014). Herein, we present data on composition and infection patterns of endoparasites associated with O. carvalhoi from a relictual forest in the Brazilian state of Ceará. MATERIAL AND METHODS Tis study was carried out in the municipality of Guaramiranga, located at the Baturité massif, northern Ceará state, Brazil. Its mountainous relief, dissected forming small valleys, covered by rainforest vegetation, is classifed as a remnant of the Atlantic Forest. Altitude. The annual rainfall has an average of 1737.5 mm/year, with a rainy season from January to May. Its average temperature ranges from 24º to 26ºC (Vale & Soares, 2006; IPECE, 2017).Field work was taken in three sampling points: Guaramiranga farm (04º15’54,91” S; 38º56’00,24” W), Álvaro farm (04º17’17,29” S; 38º57’00,05” W), Riacho Fundo farm (04º15’42,86” S; 38º55’07,57” W), Parque das Trilhas (04º16’13,63” S; 38º56’19,10” W), Vale das Nuvens (4º16’05,10” S; 38º54’54,96” W) and Pernambuquinho farm (04º12’19,73” S; 38º 57’37,70” W) . We used specimens collected from 1994 to 2022, that are deposited in the Herpetological Collection of the Universidade Federal do Ceará (CHUFC-A 2.918, 3.738-39, 3.740-41, 3.769-73, 3.799, 3.864, 3.875-76, 9.961-70, 10.275-81, 10.283-10.288, 10.999), and Herpetological Collection of the Universidade Regional do Cariri (URCA-H 16.041), Ceará state, Brazil. Tirty-nine individuals of O. carvalhoiwere collected by hand through active and auditory searches (Bernarde, 2012). Te specimens were placed in separate containers and euthanized by a lethal injection of lidocaine (CFMV, 2013), fxed with 10% formalin according to Callefo (2002). Morphometric measurements of hosts were taken with a digital caliper Mitutoyo® (precision 0.01 mm). Te specimens were necropsied with a midventral incision and all organs and coelomic cavity were searched for helminths. Parasites found were preserved in 70% ethanol, fxed according Amato et al.(1991) and Andrade (2000), and mounted in temporary slides for identifcation. For identifcation, we follow Yamaguti (1961), Sprent (1978), Vicente et al.(1991), Anderson (2000) and Gibbons (2010), as well recent descriptions. Slides were analyzed under microscope with computerized image analysis system. Voucher helminths were deposited at Coleção Parasitológica of the Universidade Federal do Ceará. Parasitological descriptors (prevalence, mean intensity of infection and abundance) were calculated according to the specifcations of Bush et al. (1997).To evaluate the impact of host size on infection intensity, we employed linear regression. Diferences in the prevalence and intensity of infection between genders were assessed using the chi-square method. Te analyses were conducted on the R platform, “Commander R” package (R CORE TEAM, 2022, version 4.2.0) (R Foundation, 2017).Ethic aspects: Tis study was approved by the Ethics Committee on Animal Use of the Federal University of Ceará (CEUA-UFC) under the protocol 6314010321.
image/svg+xml250Neotropical Helminthology (Lima). Vol. 17, Nº2, jul - dic 2023Feitosa Quirino et al.RESULTS We examined 39 specimens of O. carvalhoi, being 16 males and 23 females, which are infected with at least one endoparasite species (overall prevalence 84,97%). We found 5.681 helminths, with mean intensity of infection 153.54 ± 32.38. Parasite community was composed by 11 species: Aplectana hylambatis(Baylis, 1927), Oswaldocruzia mazzaiTravassos, 1935, Cosmocerca brasiliense Travassos, 1925,Gorgoderina parvicava Travassos, 1922,Physalopterasp., Raillietnemasp., Rhabdiassp., Strongyloidessp., Parapharyngodon sp., Oxyascaris sp.and Ochoterenellasp., besides Cosmocercidae larvae. Te most prevalent taxa were A. hylambatis(76.67%) and O. mazzai(79.49%), being the frst one with the higher abundance (102.49) and mean intensity of infection (121.12). G. parvicava, Strongyloidessp. and Parapharyngodon sp. have the lower prevalence (2.56%) (Table 1).Table 1.Hosts, number of helminths (NH), mean abundance (MA), mean intensity of infection (MII) with standard error (SE), intensity of infection amplitude (IIA) and infection site (IS) of helminths associated with Odontophrynus carvalhoi.HelminthNHMA±SEMII±SEIIAISAplectana hylambatis3692102.49±24.57121.12±27.871-725B/E/IG/IDOswaldocruzia mazzai61879.49±15.8519.94±9.941-312E/IG/IDCosmocerca brasiliense110.28±0.25.5±0.55-6IDGorgoderina parvicava50.13±0.1355BPhysalopterasp.2195.62±1.9215.64±4.281-52E/IG/IDOxyascaris sp.20.05±0.0411IG/IDRaillietnemasp.2737.0±6.491.0±79.251-249B/IDRhabdiassp.1393.56±0.836.04±1.151-21P/F/EStrongyloidessp.20.05±0.0522IDParapharyngodon sp.10.03±0.0311IGOchoterenellasp.41410.62±5.3418.82±9.171-205CAV/B/F/C/IDMean richness was 3.41 ± 0.23 helminths/hosts, and the higher richness (S=6) was found in a single host. Tere was an infuence of host size on mean intensity of infection (r=0.48; p<0.05; n=39). As for the infuence of size on the average intensity of infection between the sexes, we can observe that were signifcant for males and females (r=0.56; p<0.05; n=16, r=0.47; p<0.05; n=23, respectively).DISCUSSION Helminths community associated with amphibians are characterized by generalist species (Aho, 1990). Low host specifcity, combined with the general lack of studies with Neotropical amphibian species, are often the main causes of new host records (Campião et al., 2015; Oliveira et al., 2019). In Brazil, species of Odontophrynidae still need further studies to understand what are the host specifcities and patterns of infection and abundance (Campião et al., 2014). Species of Aplectana are usually found infecting the large intestine of reptiles and amphibians, have a direct life cycle and actively infect their hosts (Travassos, 1931; Anderson, 2000; Campião et al.,2014; Lins et al.,2017). Tis genus has been already reported in four odontophrynids: Proceratophrys tupinambaPrado and Pombal, 2008 and P. boiei(Wied-Neuwied, 1824) infected by A. deliraeFabio, 1971 (Boquimpani-Freitas et al., 2001; Klaion et al., 2011), and P. cristiceps(Silva et al., 2019; Sampaio et al., 2020) and Odontophrynus americanusDuméril & Bibron, 1841, infected by A. membranosa(Lent & Freitas, 1948).Aplectana hylambatisis a generalist species that parasitizes a wide range of hosts. In the Neotropical region it has been observed in diferent countries and families of anurans such as Bufonidae, Leptodactylidae, Ceratophryidae e Microhylidae from Peru (Bursey et al., 2001); Bufonidae, Mycrohylidae, Leiuperidae, Leptodactylidae and Hylidae from Paraguay (Masi Pallares & Maciel, 1974; Baker & Vaucher, 1986); Bufonidae from Uruguay (Lent & Freitas, 1948); Bufonidae, Leptodactylidae and Leiuperidae from Argentina (Gutiérrez, 1945; Sueldo & Ramírez, 1976;
image/svg+xml251Helminths Infecting the Carvalho’s EscuerzoNeotropical Helminthology (Lima). Vol. 17, Nº2, jul - dic 2023Ramírez et al.,1979; Baker, 1980; González & Hamann, 2006, 2010). In Brazil, it was reported infecting Hylidae, Microhylidae and Leptodactylidae at São Paulo and Mato Grosso do Sul states (Campião et al., 2016; Aguiar et al., 2021).CosmocercaDiensing, 1861, is a genus that pursues a wide geographic distribution and is commonly found as parasites of amphibians (Navarro et al., 1988). Cosmocerca brasiliensecan infect amphibian hosts by ingestion of an infective larvae (L3) or penetration through the skin (Goldberg at al.,2002a). After infection, larvae migrate to the host intestine, reaching sexual maturity. Tis species has been recorded parasitizing many hosts in South America, such as Ecuador (Dyer & Altig, 1976; Mcallister et al.,2010a), Guiana (Mcallister et al.,2010b) and Peru (Bursey et al.,2001). In Brazil, it was reported by Travassos (1925); Vicente et al.(1991); Boquimpani-Freitas et al.(2001); Martins & De Fabio (2005); Goldberg et al. (2007); Santos et al.(2013), (2016); Aguiar et al.(2014); Klaion (2011); Campião et al. (2014); Oliveira et al.(2022).Oswaldocruzia mazzaiinfect several anurans (e.g., Campião et al., 2014; Teles et al., 2015; Alcantara et al., 2018; Oliveira et al., 2019), but we reported for the frst time the infection of this nematodein the genus Odontophrynus. Te great host diversity recorded for O. mazzaiis related to the direct life cycle and the simple mode of transmission that can occur by ingestion of eggs or larval penetration of the host’s skin (Anderson, 2000).Te genus GorgoderinaLooss, 1902, includes 57 species, all of them described as parasites of the bladder of anurans and salamanders. In Brazil, seven species of this genus are known. G. parvicavais well distributed in Neotropical region and has already been reported as parasites of several anurans in Brazil, such as Leptodactylus chaquensis,Cei, 1950, L. labyrinthicus, (Spix, 1824), L. latrans(= L. ocellatus) (Stefen, 1815), L. pentadactylus, (Laurenti, 1768), Rhinella crucifer, (Wied-Neuwied, 1821), R. diptycha, (Cope, 1862), R. icterica, (Spix, 1824), R. marina, (Linnaeus, 1758), Pseudis paradoxa(Linnaeus, 1758), and Pristimantis relictusRoberto et al.,2022, (Alcantara et al., 2022; Oliveira et al., 2022).Nematodes of the genus Physalopterahave a worldwide distribution and have been recorded in several terrestrial vertebrates, including felines (Ogassawara, 1986), rodents (Tung et al., 2009), lizards (Da Silva et al., 2008, Cabral et al., 2018) and anurans (Da Graça et al.,2017). In amphibians, this parasite is usually found in the larval stage, making it difcult to identify at the species level, and it is suggestive that these amphibians are not defnitive hosts. Although there is not enough data on their life cycle, nematodes of this genus are known to utilize insects during their intermediate phase (Anderson, 2000). Additionally, the acquisition of Physalopteraby anuran hosts occurs through the ingestion of infected insects, mainly Orthoptera (Klaion et al., 2011).Raillietnemasp. is known to have a direct life cycle and transmission that occurs via ingestion or penetration of larvae through the skin (Anderson, 2000). Studies have reported the presence of this parasite infecting P. aridus(Teles et al., 2017), as well as infecting lizards. Te species R. spectans(Bursey et al.,1998), are the most frequent in anurans (Vicente et al., 1991; Teles et al., 2015). Te specifc identifcation of the Rhabdiasfound here was not possible, because there is high morphological similarity, which makes molecular data useful in species recognition (Müller et al., 2018). Nematodes of this genus are lung parasites, commonly infecting amphibians and reptiles, by direct transmission and infection occurs by active penetration into the skin of hosts (Langford & Janovy, 2009; Kuzmin et al., 2015). Although the life cycle of Strongyloidesis not known, it can be considered a direct or indirect cycle, the frst being the most common (Santos et al., 2010). Infection occurs on land through skin penetration or ingestion of infected prey (Mati & Melo, 2014; Sulieman et al., 2015). Although there are several records of this nematode infecting amphibian species (Campião et al.,2014; Sulieman et al.,2015; Mascarenhas et al.,2021), this is the second record of the genus Strongyloidesacting as a parasite for species of the Odontophrynidae family and the frst for O. carvalhoi.ParapharyngodonChatterji, 1933, is a genus of parasitic nematodes with monoxenic cycle (Anderson, 2000). Tese have already been registered in amphibian species (Pereira et al.,2017), being commonly found in reptiles (Avila & Silva, 2010). Ochoterenella is reported from South and Central America, with several species known only from females and larvae. Te genus is remarkably diverse in bufonids such as Rhinella marina(Linnaeus, 1758) (Travassos, 1929; Caballero, 1944; Bain & Prod’Hon, 1974; Bain et al.,1979; Esslinger 1986, 1987, 1988a, b, 1989), while only two of 15 species have been described for Leptodactylidae with few records for hylids (Lima et al.,2012).
image/svg+xml252Neotropical Helminthology (Lima). Vol. 17, Nº2, jul - dic 2023Feitosa Quirino et al.Oxyascarisgenus was reported parasitizing the digestive system of anurans and reptiles from Paraguay, Argentina and Costa Rica. In Brazil, species of this genus were recorded in Bufonidae, Hylidae, Leptodactylidae, Phyllomedusidae and Cycloramphidae anuran families (Freitas, 1958; Baker & Vaucher, 1985; Luque et al.,2005; Bursey & Goldberg, 2007; Lux-Hoppe et al., 2008; González & Hamann, 2008; Goldberg & Bursey, 2008a; Goldberg & Bursey, 2008b; González, 2009; Bursey & Goldberg, 2010; Klaion et al.,2011; Santos et al., 2013; Campião et al.,2014, 2016; De Sena et al., 2018; Felix-Nascimento et al., 2020). Despite being recorded only in Neotropical hosts, very little is known about its life cycle and how transmission occurs. In recent years, there is an increase in studies of helminths associated with amphibians (Campião et al.,2016a, b; Lins et al.,2017; Teles et al.,2017; Leivas et al.,2018; Alcantara et al., 2018; Oliveira et al.2019; Mascarenhas et al.,2021), with new host records being quite common. In this study, we present new records of parasite species for the species of O. carvalhoi, contributing to the knowledge of the parasitic fauna of the Neotropical region.ACKNOWLEDGEMENTSTe authors thank Conselho Nacional de Desenvolvimento Científco e Tecnológico (CNPq) for providing a research fellowship to (R.W.A., 303622/2015-6; 305988/2018-2; 307722/2021-0); To Coordenação de Aperfeiçoamento de Pessoal de Nível Superior- Brasil (CAPES) and Fundação Cearense de Apoio Científco e Tecnológico FUNCAP for the doctoral scholarship (Finance Code 001). To Instituto Chico Mendes de Conservação da Biodiversidade – ICMBio for collecting permits (SISBIO 29613).Author contributions: CRediT (Contributor Roles Taxonomy) TFQ= Tatiana Feitosa Quirino DBO= Dalilange Batista-Oliveira MCS= Matheus Calixto Saldanha RWA= Robson Waldemar ÁvilaConceptualization: TFQ, DBO, MCS, RWAData curation: TFQ, DBO, MCS, RWAFormal Analysis: RWAFunding acquisition: TFQ, DBO, RWAInvestigation: TFQMethodology: TFQ, DBO, MCSProject administration: TFQResources: TFQ, DBO, MCS, RWASoftware: TFQ, DBO, MCS, RWASupervision: RWAValidation: TFQ, DBO, MCS, RWAVisualization: TFQ, DBO, MCS, RWAWriting – original draft: TFQ, DBO, MCSWriting – review & editing: RWABIBLIOGRAPHIC REFERENCES Aguiar, A., Morais, D.H., Silva, L.A.F., & Silva, R.J. (2015). Te frst report of Aplectana hylambatis(Nematoda: Cosmocercidae) associated with Dermatonotus muelleri(Anura: Microhylidae) from Brazil. Herpetological Review, 46(3), 336-338.Aho, J.M. (1990). Helminth communities of amphibians and reptiles: comparative approaches to understanding patterns and process. In Esch, G.W., Bush, A.O. & Aho, J.M. (Eds.). Parasite communities: patterns and process.London: Chapman and Hall. p. 157-199. http://dx.doi.org/10.1007/978-94-009-0837-6_7.Alcantara, E.P., Ferreira-Silva, C., Silva, L.A.F., Lins, A.G.S., Ávila, R.W., Morais, D.H. & Silva, R.J. (2018). Helminths of Dermatonotus muelleri(Anura: Microhylidae) from Northeastern Brazil. Journal of Parasitology. 104(6), 550-556.Alcantara, E. P., Úngari, L. P., Muller, M. I., Ferreira-Silva, C., Emmericha, E., O’Dwyera, L. H., & Silva, R. J. (2022). Phylogenetic position of Gorgoderinaparvicava Travassos, 1922 (Digenea: Gorgoderidae), a parasite of Leptodactyluslabyrinthicus(Spix, 1824) (Anura: Leptodactylidae) in Brazil. Brazilian Journal of Biology, 82. https://doi.org/10.1590/1519-6984.263745.
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