65 Endoparasites of anurans in the seven cities national park Neotropical Helminthology, Vol. 19, Nº1, jan - jun 2025 Neotropical Helminthology Neotropical Helminthology, 2025, vol. 19 (1), 65-74 ORIGINAL ARTICLE / ARTÍCULO ORIGINAL NEMATODES INFECTING PHYLLOPEZUS POLLICARIS (SPIX, 1825) (REPTILIA: PHYLLODACTYLIDAE) IN THE PALMARES NATIONAL FOREST, PIAUÍ STATE, BRAZILNEMATÓDEOS INFECTANDO PHYLLOPEZUS POLLICARIS (SPIX, 1825) (REPTILIA: PHYLLODACTYLIDAE) NA FLORESTA NACIONAL DE PALMARES, ESTADO DO PIAUÍ, BRASILNEMATODOS QUE INFECTAN A PHYLLOPEZUS POLLICARIS (SPIX, 1825) (REPTILIA: PHYLLODACTYLIDAE) EN EL BOSQUE NACIONAL DE PALMARES, ESTADO DE PIAUÍ, BRASIL Brenda Milena Bezerra Costa 1 , Kássio de Castro Araújo 1 , Cristiana Ferreira-Silva 2 , Ronildo Alves Benício 3 , Jussara da Conceição Eduardo 1 , Samuel Lopes Pereira 1 &Etielle Barroso de Andrade 1* ISSN Versión Impresa 2218-6425 ISSN Versión Electrónica 1995-1403 DOI: https://dx.doi.org/10.62429/rnh20251911910 Universidad Nacional Federico Villarreal Volume 19, Number 1 (jan - jun) 2025 Este 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. ABSTRACT Parasitological studies are important to understand the host-parasite interaction, providing information on the coevolution, biogeography, morphology, and diet of the species. T e present study describes the endoparasite fauna of the lizard Phyllopezus pollicaris (Spix, 1825) in the Palmares National Forest, Piauí State, Brazil, a tropical forest covered by transitional vegetation between the Caatinga and Cerrado biomes. Furthermore, we investigated the inf uence of 1 Grupo de Pesquisa em Biodiversidade e Biotecnologia do Centro-Norte Piauiense -BIOTECPI, Instituto Federal de Educação, Ciência e Tecnologia do Piauí - IFPI Campus Pedro II, 64255-000, Pedro II, Piauí, Brasil. 2 Núcleo Regional de Of ologia, Universidade Federal do Ceará - UFC, Bloco 905, Centro de Ciências, Campus PICI, 60455-760,Fortaleza, Ceará, Brasil. 3 Laboratório de Herpetologia e Parasitologia de Animais Silvestres, Universidade Federal do Piauí - UFPI, Campus Picos, 64607-670, Picos, Piauí, Brasil. * Corresponding author: brendamilena06@gmail.com Brenda Milena Bezerra Costa: https://orcid.org/0009-0004-3037-2769Kássio de Castro Araújo: https://orcid.org/0000-0003-4091-8521Cristiana Ferreira-Silva: https://orcid.org/0000-0002-3087-2614Ronildo Alves Benício: https://orcid.org/0000-0002-7928-2172Jussara da Conceição Eduardo: https://orcid.org/0009-0005-1233-0220Samuel Lopes Pereira: https://orcid.org/0009-0008-7618-4535Etielle Barroso de Andrade: https://orcid.org/0000-0002-5030-1675
66 Neotropical Helminthology, Vol. 19, Nº1, jan - jun 2025 Bezerra Costa et al. lizards’ size and sex on endoparasite infection patterns. We analyzed 23 individuals of P. pollicaris , of which 18 were infected by at least one helminth, belonging to the Nematoda phylum: Spauligodon oxkutzcabiensis (Chitwood, 1938), Strongyloides sp . , and Parapharyngodon sp. Overall, S. oxkutzcabiensis showed the highest prevalence (73.9%), mean infection intensity (8.64 ± 12.7), and mean abundance (6.39 ± 11.5). We observed that the lizard size was positively related to the abundance of endoparasites, while the sex of individuals had a poor efect on parasitological descriptors. Tis study contributes information about the endoparasite fauna associated with P. pollicaris , composed exclusively of nematodes. As a pioneering study, it can be considered the starting point for parasitological studies involving lizards in Piauí State, Northeastern Brazil. Keywords : Geckos – helminths – lizards – parasites RESUMO Estudos parasitológicos são importantes para se compreender a relação parasito-hospedeiro, fornecendo informações sobre a coevolução, biogeografa, morfologia e dieta das espécies. O presente estudo descreve a fauna endoparasitária do lagarto Phyllopezus pollicaris (Spix, 1825) na Floresta Nacional (Flona) de Palmares, Piauí, Brasil, uma foresta recoberta por vegetações de transição entre o Cerrado e a Caatinga. Além de investigar a infuência do sexo e do tamanho dos lagartos nos padrões de infecções. Analisamos 23 exemplares de P. pollicaris dos quais 18 estavam infectados por pelo menos um helminto, pertencente a três espécies de nematódeos: Spauligodon oxkutzcabiensis (Chitwood, 1938), Strongyloides sp . e Parapharyngodon sp. No geral, a maior prevalência (73,9), intensidade média de infecção (8,64 ± 12,7) e abundância média (6,39 ± 11,5) foi de S. oxkutzcabiensis . Nós observamos que o tamanho dos lagartos está positivamente relacionado com a abundância de parasitos, enquanto que o sexo do indivíduo teve pouco efeito nos descritores parasitológicos. Este estudo contribui com informações sobre a fauna de endoparasitas associada a P. pollicaris , sendo composta exclusivamente por nematoides. Por ser pioneiro, pode ser considerado o ponto de partida para estudos parasitológicos envolvendo lagartos no estado do Piauí, Nordeste do Brasil. Palavras-chave : Gekkos – helmintos – lagartos – parasitos RESUMEN Los estudios parasitológicos son importantes para entender la relación parásito-huésped, proporcionando información sobre la coevolución, biogeografía, morfología y dieta de la especie. El presente estudio describe la fauna endoparásita del lagarto Phyllopezus pollicaris (Spix, 1825) en el Bosque Nacional de Palmares, Estado de Piauí, Brasil, un bosque cubierto por vegetación de transición entre el Cerrado y la Caatinga. Además de investigar la infuencia del sexo y tamaño de las lagartijas en los patrones de infección. Se analizaron 23 ejemplares de P. pollicaris , de los cuales 18 estaban infectados por al menos un helminto, pertenecientes a tres especies de nematodos: Spauligodon oxkutzcabiensis (Chitwood, 1938), Strongyloides sp. y Parapharyngodon sp. En general, la mayor prevalencia (73,9), intensidad media de la infección (8,64 ± 12,7) y abundancia media (6,39 ± 11,5) fue de S. oxkutzcabiensis . Observamos que el tamaño de la lagartija se relaciona positivamente con la abundancia de parásitos, mientras que el sexo del individuo tuvo poco efecto en los descriptores parasitológicos. Este estudio contribuye con información sobre la fauna endoparásita asociada a P. pollicaris , que está compuesta exclusivamente por nematodos. Al ser pionero, puede considerarse el punto de partida para los estudios parasitológicos que involucran lagartos en el estado de Piauí, en el noreste de Brasil. Palabras clave : Gekkos – helmintos – lagartos – parásito
67 Nematodes infecting Phyllopezus pollicaris Neotropical Helminthology, Vol. 19, Nº1, jan - jun 2025 INTRODUCTION Parasites are organisms that need other living organisms to survive, using the host as a food source and intermediate or permanent home (Windsor, 1995). Tey are classifed as ectoparasites when they live outside or in the host’s pits (e.g., ticks) or endoparasites residing inside the host (e.g., nematodes; Rey, 2018). Regarding endoparasites, they are classifed as macro or microparasites, for example, arthropods, plathelminths, nematodes, fungi, and bacteria (Pantoja et al ., 2015). Parasitological studies provide information about the coevolution, biogeography, morphology, and diet of species since parasitic infections signifcantly afect their hosts (Poulin, 2021). Furthermore, endoparasites play an important role in the ecosystems acting as environmental bioindicators (Poulin, 1999; Zargar & Chishti, 2022).Lizards are interesting ecological models for parasitological studies because of their well-resolved taxonomy and for acting as intermediate and defnitive hosts of diverse endoparasite species (Wilson & Carpenter, 1996). Tis interaction is usually associated with the host forage mode and diet, in which they are infected directly by larvae cutaneous penetration or by ingestion of infected prey (González et al. , 2021; Dantas et al ., 2024).Lizards are one of the most diverse vertebrate groups, with 7,724 species registered in the world (Uetz et al ., 2024). Morphophysiological adaptations and the water- independence to reproduce contributed to the lizard diversifcation worldwide (Hickman et al ., 2013). Tere are 292 lizard species in Brazil, of which 14 belong to the Phyllodactylidae family (Guedes et al ., 2023). Tese lizards are characterized by having smooth skin with the presence of granules, well-developed limbs, non-existent eyelids, and adhesive lamellae that they use to attach themselves to surfaces (Rocha, 1994; Gamble et al ., 2012; Cacciali et al ., 2018).Among the Phyllodactylidae, Phyllopezus pollicaris (Spix, 1825) (Reptilia: Phyllodactylidae) is a mid-size Gecko reaching up to 20 cm in total length, characterized by having a depressed body, fattened head, snout with large granules, well-developed limbs, and distinct coloring between juveniles and adults (Rodrigues, 1985; Vanzolini et al .,1980). Tis species is widely distributed in Brazil, being commonly recorded in the Caatinga and Cerrado biomes (Gamble et al ., 2012; Guedes et al. , 2023). It is a nocturnal lizard that inhabits rocky outcrops, fallen tree trunks, leaf litter, and also urban areas. In addition, it is a “sit-and-wait” forager having an insectivore diet (Silva & Araújo, 2008; Gamble et al ., 2012; Carvalho et al ., 2014). Some studies already reported endoparasites infecting P. pollicaris , for exemple, nematodes and cestodes were found parasitizing this lizard in the Estação Ecológica de Aiuaba (Lima et al ., 2017) and in an urban area of Crato municipality (Sousa et al ., 2014), both in Ceará State, Northeastern Brazil. In these studies, Spauligodon oxkutzcabiensis (Chitwood, 1938) had the highest prevalence and mean infection intensity. Specifcally in Piauí State, there is one only record of Raillietiella mottae (Almeida, Freire & Lopes, 2008) parasitizing this lizard (Sousa et al., 2010). Terefore, due to the lack of parasitological studies dealing with lizard species in Piauí State, the present study aimed to investigate the endoparasites associated with P. pollicaris in the Palmares National Forest, Piauí State, Northeastern Brazil, besides testing how the size and sex of the individuals infuence the infection patterns. MATERIAL AND METHODSStudy area Te present study was conducted in the Palmares National Forest (Fig. 1), situated in the municipality of Altos, Piauí State, Northeastern Brazil (05°03’25.90”S, 42°35’34.04”W, elev. 160 m). It is a small federal conservation unit comprising 170 ha, created by decree S/N of 21 February 2005, that aims to maintain and protect wild animals and native forest species in Piauí (ICMBio, 2022). It is a seasonal semideciduous (ecotonal) forest inserted in a transition area between the Cerrado and Caatinga biomes (IBGE, 2019). Te climate is tropical with two well-defned seasons (rain and dry seasons), annual rainfall ranging from 1200 to 1400 mm, and annual mean temperature of 28º C (Brandão et al. , 2020; ICMBio, 2022; INMET, 2024). Sampling We searched for individuals of P. pollicaris using visual active searches in microhabitats usually used by them, such as trunks, trees, and sandstone outcrops (see Recoder et al. , 2012; Gonçalves et al. , 2019). We carried out two samplings of fve consecutive days in February and March, totaling 10 collection days. Four researchers conducted the samplings in the morning (9 to 12 h) and night (18 to 23 h), totaling 320 hours of sampling efort. We euthanatized the collected lizards following the guidelines established by the National Council for the Control of Animal Experimentation in accordance with Law 11.794 of 2008, article 14 § 1º and with Normative
68 Neotropical Helminthology, Vol. 19, Nº1, jan - jun 2025 Bezerra Costa et al. Resolution 37, February 15, 2018 (Concea, 2018), and the Brazilian Guide for the Production, Maintenance or Use of Animals in Teaching or Scientifc Research Activities (Jared et al. , 2023). Figure 1. Location map of the Palmares National Forest, Piauí State, Northeastern Brazil, highlighting the main visiting trails. Parasitological procedures In the lab, we measured the snout-vent length of each collected individual using a digital caliper (precision of 0.01 mm), and we necropsied the individuals with a ventral incision and removed their organs with the help of a stereomicroscope. Firstly, we analyzed the presence of endoparasites in the oral cavity and, thereafter, the following organs: digestive tract, lungs, heart, liver, and kidneys (Amato et al ., 1991). Te collected endoparasites were fxed in 10% formalin, preserved in 70% alcohol, and housed in the Helminthological Collection of the Universidade Federal do Ceará, Fortaleza, Brazil. Data analyses According to Bush et al . (1997), we used the following parasitological descriptors: richness (total number of endoparasite species), abundance (total number of endoparasites), mean abundance (total number of a given species of parasite in an individual/number of hosts examined), prevalence (number of infected hosts/total number of hosts analyzed), and mean infection intensity (total number of a species of endoparasites/number of hosts infected by the same parasite). After that, we used the abundance data to investigate how the lizard sizes infuence the endoparasite infection. For this, we used the non-parametric regression of Kernel (Nadaraya, 1964) because our data did not follow the assumptions for the linear regression test. We used the R packages Vegan (Oksanen et al. , 2024) and ggplot2 (Wickham, 2016) to perform this analysis. E thic aspects : All the procedures used in this work comply with the ethical standards of the relevant national and institutional guidelines on the care and use of laboratory animals. Collection authorization Chico Mendes Institute for Biodiversity Conservation - ICMBio (#86665-2). Voucher specimens were housed in the Biological Collection of Instituto Federal de Educação, Ciência e Tecnologia do Piauí, Campus Pedro II (CBPII).
69 Nematodes infecting Phyllopezus pollicaris Neotropical Helminthology, Vol. 19, Nº1, jan - jun 2025 RESULTS We collected 23 P. pollicaris individuals (12 males and 11 females), of which 18 had at least one helminth parasite (total prevalence = 78.2%). Overall, we found 149 nematodes with a mean abundance of 6.47 endoparasites per individual and 8.27 mean infection intensity. Te highest endoparasite abundance was found in the larger intestine (139 nematodes), whereas 10 were recorded in the stomach and fve in the small intestine (Table 1). Table 1. Endoparasites associated with Phyllopezus pollicaris in the Palmares National Forest, Piauí State, Northeastern Brazil, with the respective number of helminths recorded (NH), prevalence (P), mean infection intensity (IM), mean abundance (MA), minimum and maximum number of parasites per individual (Min-Max) and site of infection (IG). EndoparasitosNHP%IM±SDMA±SDMin-MaxSI Spauligodon oxkutzcabiensis 14773.98.64 ± 12.76.39 ± 11.51-41ST, LI, SI Strongyloides sp.14.3410.04 ± 0.201-1SI Parapharyngodon sp.14.3410.04 ± 0.201-1LI TOTAL 14978.28.27 ± 12.46.47 ± 11.51-41 Legends: standard deviation (SD), stomach (ST), large intestine (LI) and small intestine (SI) . Figure 2. Nematode species parasitizing Phyllopezus pollicaris in the Palmares National Forest: A) Spauligodon oxkutzcabiensis , B) Strongyloides sp., and C) Parapharyngodon sp. Phyllopezus pollicaris was exclusively parasitized by nematodes belonging to three species: S. oxkutzcabiensis , Strongyloides sp . , and Parapharyngodon sp. (Fig. 2). Among these, S. oxkutzcabiensis had highest prevalence, abundance, and mean infection intensity. Te other two nematode species parasitized a single male individual. Additionally, we observed that males had the highest abundance (NH = 89), mean abundance (MA = 7.42), prevalence (P = 83.3), and mean infection intensity (IM = 8.90) than females (NH = 60, MA =5.45, P = 72.3, IM = 7.50).It is interesting to note that the abundance of endoparasites was positively related to the lizard size (F = 4.481, P = 0.025), thus, larger individuals tended to be more parasitized (Fig. 3). DISCUSSION We found a high abundance of helminths parasitizing P. pollicaris belonging to three species, composed of nematodes exclusively. Tese helminths are usually found parasitizing reptiles (Anderson, 2000; Lacerda et al ., 2023), and exclusively nematodes infection was already reported for other lizards such as Tropidurus hispidus (Spix, 1825) and T. semitaeniatus (Spix, 1825), and even Geckos such as Hemidactylus mabouia (Moreau de Jonnés, 1818) (Ávila et al. , 2012; Anjos et al., 2013).
70 Neotropical Helminthology, Vol. 19, Nº1, jan - jun 2025 Bezerra Costa et al. Among the Phyllopezus species, our results support the fndings this lizard tend to be predominantly parasitized by nematodes, mainly for S. oxkutzcabiensis in the Caatinga biome (Ávila et al. , 2012; Sousa et al ., 2014; Lima et al. , 2017). Te high infections taxa of S. oxkutzcabiensis may be associated with its monoxenic life cycle, in which parasitic infection can occur through the ingestion of eggs (Anderson, 2000). Tis helminth had the highest abundance and prevalence in the present study, and it is commonly reported parasitizing other geckos in diferent Brazilian biomes, as Cerrado, Caatinga, and Mata Atlantica (Ávila & Silva, 2010; Ávila et al ., 2012; Ávila et al ., 2010; Araujo et al ., 2020). It is noteworthy to note that the host’s biological characteristics also infuence the parasitic infection patterns (Aho, 1990).We found only one Strongyloides sp. specimen parasitizing the larger intestine of a single male individual of P. pollicaris . Te genus Strongyloides can be usually found in the gastrointestinal tract of reptiles, birds, amphibians, and mammals (Little, 1966; Roca & Hornero, 1992). Tese parasites have a complex life cycle with two phases: in the free-living phase, male and female individuais are found that develop through indirect development and exchange gametes (sexual or heterogonic reproduction), and in the parasitic phase, parthenogenetic females are found only that produce eggs without the need for a male (asexual or homogonic reproduction) (Viney & Lok, 2007). Tis is the frst report of this genus parasitizing P. pollicaris , despite this nematode being frequently reported to infect reptiles (Ávila & Silva, 2010; Mati et al ., 2013; Mesquita et al. , 2020). Because we only found one female with a broken posterior region, we were unable to count the number of turns the posterior ovary makes around the intestine, a characteristic crucial to species identifcation (Little, 1966). As a result, we were unable to identify this helminth to the species level. Parapharyngodon is a nematode genus widely distribut-ed and commonly recorded infecting insectivore reptile species (Adamson, 1981), including lizards such as P. pollicaris, Ameiva ameiva (Linnaeus, 1758), and T. semi-taeniatus (Lacerda et al ., 2023). Once this helminth has a monoxenic lifecycle (Anderson, 2000), host infection probably occurs during foraging activity, where this lizard accidentally ingested the eggs of infectious parasites that were attached to its prey or in the environment. However, further investigations are necessary to prove the validity of this proposal. Parapharyngodon sp. was not identifed to species level because only a single female was record-ed, and the identifcation of Parapharyngodon species is carried out based mainly on the reproductive characteris-tics of males, such as morphology and size of the spicule, number and arrangement of caudal papillae, presence or absence of genital cone, and shape of the anterior lip of the cloaca. Figure 3 . Relationship between lizard size (SVL) and abundance of parasites associated with Phyllopezus pollicaris in the Palmares National Forest, Piauí, Northeastern Brazil. We used logarithmized values for a better visualization.
71 Nematodes infecting Phyllopezus pollicaris Neotropical Helminthology, Vol. 19, Nº1, jan - jun 2025 We observed that larger lizards tended to be more infected (highest endoparasite abundance). Based on the species-area hypothesis (see Tjørve et al. , 2008), it is expected that larger hosts will have more space and high niche diversity to house endoparasites (Anjos et al. , 2012; Kamiya et al. , 2013; Campião et al ., 2015). Tis pattern was already reported in lizards, including P. pollicaris in an urban area in Northeastern Brazil (Sousa et al ., 2014). However, further studies are still needed to understand the main drivers of endoparasite infections in lizard species. Overall, this study contributes to the knowledge of the parasitic fauna of the lizard P. pollicaris , being the frst study carried out in the Palmares National Forest, Piauí State, Northeastern Brazil. Aspects related to seasonality, foraging activity and feeding habits of hosts infuence endoparasite infections, however, little is known about how parasitological descriptors are afected by the environmental characteristics of the Palmares National Forest. Terefore, we reinforce the need of more parasitological studies in this conservation unit aiming to understand the host-endoparasite in lizard species of Piauí State. ACKNOWLEDGMENTS RAB and KCA thank Conselho Nacional de Desenvolvimento Científco e Tecnológico (CNPq) and Fundação de Amparo à Pesquisa do Estado do Piauí (FAPEPI) for providing research fellowship (Processes: 301239/2022-3; 150013/2023-0). CFS thank to CNPq (#150125/2023-2) and thank Fundação Cearense de Apoio ao Desenvolvimento Científco e Tecnológico (FUNCAP) (#FC3-0198-00006.01.00/22) for research funding. We thank Andreia Andrade, Douglas Oliveira, and João Lucas Ferreira for feldwork support. We also thank the manager of the Palmares National Forest, Gaspar da Silva Alencar, for logistical support. Author contributions: CRediT (Contributor Roles Taxonomy)BMBC = Brenda Milena Bezerra Costa KCA = Kássio de Castro Araújo CFS = Cristiana Ferreira-Silva RAB = Ronildo Alves Benício JCE = Jussara da Conceição Eduardo SLP = Samuel Lopes Pereira EBA = Etielle Barroso de Andrade Conceptualization: BMBC, KCA Data curation: BMBC, EBA Formal Analysis: BMBC, KCA Funding acquisition: RAB Investigation: BMBC, KCA, RAB, EBA Methodology: BMBC, KCA, RAB, EBA, CFS, JCE, SLP Project administration: KCA, EBA Resources: RAB, EBA Software: KCA, EBA Supervision: KCA, RAB, EBA Validation: KCA, RAB, EBA Visualization: BMBC, KCA, RAB, EBA Writing – original draft: BMBC, JCE, SLP, CFS Writing – review & editing: KCA, RAB, EBA, CFS BIBLIOGRAPHIC REFERENCES Adamson, M. L. (1981). Parapharyngodon osteopili n . sp. (Pharyngodonidae: Oxyuroidea) and a revision of Parapharyngodon and Telandros. Systematic Parasitology, 3 , 105-117.Aho, J. M. (1990). Helminth communities of amphibians and reptiles: comparative approaches to understanding patterns and processes . In: Esch, G. W., Busch, A. O. & Aho, J. M. (eds). Parasite communities: patterns and processes. Chapman and Hall, New York, pp. 157-195.Amato, J. F. R., & Amato, S. B. (eds). (2010). Técnicas gerais para coleta e preparação de helmintos endoparasitos de aves. Technical Books, pp. 369 - 393.Amato, J. F. R., Boeger, W. P., & Amatos, B. (1991). Protocolos para laboratório: coleta e processamento de parasitos de pescado . Imprensa Universitária, Universidade Federal Rural do Rio de Janeiro.
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