Herpetology Notes, volume 13: 235-239 (2020) (published online on 11 March 2020)

Natural history notes and range extension of Loxopholis ferreirai (Rodrigues and Avila-Pires, 2005): a micro-endemic species from the flooded igapó forests of Rio Negro and tributaries, Brazilian Amazonia (Squamata: Gymnophthalmidae)

Sergio Marques-Souza1,*, Marco A. de Sena1, Gisele Cassundé Ferreira2, Miguel Trefaut Rodrigues1, and Tuliana O. Brunes1

Introduction Although the distribution and natural history of some semi-aquatic species is well studied (e.g., Vitt and The Amazonia forest is in one of the largest drainage Blackburn, 1991; Avila-Pires, 1995; Vitt et al., 2001), basins in the world, both in terms of area and water the knowledge about most of them is anecdotal. A volume, reaching a discharging volume of 175,000 m3/ remarkable example is the lizard Loxopholis ferreirai, s (Wohl, 2007). Due to its largely flat terrain close to locally known as “Panema’s lizard”, which means “bad rivers, large extensions are seasonally flooded by the luck” (“Panema”), as local villagers believe that, when waters of the Rio Amazonas and its tributaries (Junk, it falls on the boat, fishing will be unproductive. 2000). This so-called annual flood pulse creates selective Most of the 11 known species of Loxopholis (Pellegrino pressures to live in flooded environments, such as igapó et al., 2018) are terrestrial, living in the leaf litter of terra- and várzea forests. Given this landscape context, several firme forests (Avila-Pires, 1995; Brunes et al., 2019). lizards in Amazonia have a close association with Loxopholis ferreirai is known only from its type locality, aquatic environments, such as flooded forests, large two islands on the Arquipélago de Anavilhanas, a large and small rivers. Some genera, such as Crocodilurus, fluvial archipelago in the Rio Negro, central Amazonia, Dracaena, Kentropyx, Loxopholis, Neusticurus, subjected to annual cycles of flooding (Rodrigues and Potamites, Varzea, and Uranoscodon (Uzzell, 1965; Avila-Pires, 2005). Little is known about its distribution Vitt and Blackburn, 1991; Avila-Pires, 1995; Vitt et al., and how this species deals with the seasonally flooded 2001), present distinct levels of association with aquatic habitat. Recently, during three field expeditions to lower environments (Marques-Souza et al., 2018, Vásquez- and middle Rio Negro basin, one of the largest tributaries Restrepo et al., 2019). Among these lizards, an apparent of the Rio Amazonas, we gathered data on distribution widespread adaptation is to forage off ground, on fallen and natural history of L. ferreirai. Therefore, here we logs or tree trunks, allowing them thriving during the expand the distribution of the species to nine localities flood season (Vitt and Blackburn, 1991; Avila-Pires, on the lower and middle Rio Negro region and one 1995). Additionally, these species are commonly good tributary river. We also provide basic natural history swimmers, capable of escaping from predators during data (microhabitat usage, activity time, and oviposition predatory attempts. site) from specimens collected.

Materials and Methods Species identification was based on specimens’ external morphology (scale counts; Table 1) and 1 Departamento de Zoologia, Instituto de Biociências, taxonomic literature on Loxopholis (Uzzell and Barry, Universidade de São Paulo, Caixa Postal 11.461, São Paulo, 1971; Avila-Pires, 1995; Rodrigues and Avila-Pires, São Paulo 05508-090, Brazil. 2 Coordenação de Zoologia, Museu Paraense Emílio Goeldi, 2005; Marques-Souza et al., 2015). All specimens of Caixa Postal 399, Belém, Pará 66017-970, Brazil. L. ferreirai registered were collected and euthanized * Corresponding author. E-mail: [email protected] according to ethical guidelines (Underwood et al., 236 Sergio Marques-Souza et al.

Table 1. Scale counts of previous and new Loxopholis ferreirai specimens from sampled localities at Rio Negro and Rio Jaú, state of Amazonas, Brazil. SL = Samplig locality number; N = number of specimens analysed per locality; SAB = number of scales around midbody; Dorsals = number of transversal rows of dorsal scales; Ventrals = number of transversal rows of ventral scales; FingerTable IV 1. Scale= number counts ofof Loxopholis lamellae ferreirai under specimensfourth finger; from sampled Toe IV localities = number at Rio ofNegro lamellae and Rio under Jaú, state fourth of Amazonas, toe; Gulars Brazil. =SL number = Samplig of locality transversal number; N = number of specimens analysed per locality; SAB = number of scales around midbody; Dorsals = number of transversal rows of dorsal scales; Ventrals = number rowsof transversalof gular rowsscales; of ventral * = previously scales; Finger knownIV = number localities of lamellae (Rodrigues under fourth and finger; Avila-Pires, Toe IV = number 2005). of lamellae under fourth toe; Gulars = number of transversal rows of gular scales.

SL Locality Municipality N SAB Dorsals Ventrals Pores in males Finger IV Toe Gulars (preanal and IV femoral) 1* Lago do Prato, Arquilélago de Novo Airão 1 32 44 27 12 14 21 13 Anvilhanas 2* Ilha do Açaí, Arquipélago de Novo Airão 1 30 42 26 12 13 19 15 Anavilhanas 3 Parque Nacional do Jaú Novo Airão 23 30–33 42–45 26–28 12 14–16 19–21 13–15 4 Ilha Comunidade Santa Helena Barcelos 3 29–31 40–43 26–29 11 14–15 18–21 12 5 Igarapé do Piloto Barcelos 1 29 43 27 12 15 20 12 6 Ilha do Piloto Barcelos 5 30–31 40–44 26–28 11–12 14–15 20–21 12–13 7 Ilha das Pombas Barcelos 3 30–32 43–45 29–30 11 14–16 20 13–14 8 Ilha Paranã Uruá Barcelos 4 30–31 42–44 26–27 10–12 14–16 19–22 11–13 9 Ilha Cacauali Santa Isabel do Rio Negro 2 29 41 27 11–12 13–14 20 11 10 Poção Janatuba Santa Isabel do Rio Negro 2 30 41–44 29 11 13–14 19–21 11–12 11 Ilha Uruari Santa Isabel do Rio Negro 2 29–31 39–44 27–28 13 12–13 18–19 10–12 12 Ilha Comunidade Boa Vista Santa Isabel do Rio Negro 1 29 40 25 12 13 18 11 13 Lago Mamiará Santa Isabel do Rio Negro 1 30 42 27 10 14 19 11 Total 49 29–33 39–45 25–30 10–13 13–16 18–22 10–15

2013). A small portion of liver or muscle tissue was Acknowledgements) to Parque Nacional do Jaú (PARNA removed for DNA extraction and preserved in 100% Jaú), in the municipalities of Barcelos and Novo Airão, ethanol. The specimens were fixed with 10% formalin state of Amazonas, from the 14th of February to the 10th and preserved in 70% ethanol. Tissue samples were of March 2017 (Locality 3 in Table 2). We (SMS, MAS, deposited in the MTR Tissue Collection at the Instituto MTR, TOB, and collaborators) performed the third de Biociências, Universidade de São Paulo, state of São fieldtrip sampling several localities (localities 4, 7, 9 Paulo, Brazil. Vouchers specimens were deposited at and 10 in Table 2) in the municipalities of Barcelos and the Museu de Zoologia (MZUSP), Universidade de São Santa Isabel do Rio Negro, state of Amazonas, during Paulo, state of São Paulo, Brazil, and Museu Paraense a 26 days expedition (from the 17th of April to the 12th Emílio Goeldi (MPEG), state of Pará, Brazil. During the of May 2018). first fieldtrip (see above), we measured air temperature and relative humidity at each site where Loxopholis was Results and Discussion collected, using a portable thermohygrometer. Eggs measurements were taken with a calliper with a 0.1 mm The new localities where Loxopholis ferreirai was precision. registered were summarized in Table 2. With this data, We collected the specimens of Loxopholis ferreirai we extended the distribution of the species in c.a. 600 during three fieldtrips. We (SMS, MAS, and TOB) km northwest straight-line (Fig. 1). In Barcelos, we performed the first one from the 14th to the 23th of registered 11 specimens of L. ferreirai in two islands of February 2017 in the municipality of Barcelos, state the Arquipélago de Mariuá, being nine specimens on Ilha of Amazonas, on the middle portion of the Rio Negro do Piloto, one in Ilha Anauali (0.9303°S, 62.8670°W; (localities 5, 6 and 8 in Table 2). The city of Barcelos Datum WGS84; 33 m elevation), which was not is in front of the Arquipélago de Mariuá, one of the collected as it escaped during capture attempt, and one largest fluvial archipelagos in the world together in Igarapé do Piloto, which is situated on the right bank with Anavilhanas, located in the lower portion of of the Rio Negro (Table 2). This latter specimen was the Rio Negro. The second fieldtrip was performed found in syntopy with Loxopholis percarinatum (Müller, by Pedro L.V. Peloso (PLVP) and collaborators (see 1923) at Igarapé do Piloto. As far as we know, this is Natural history notes and range extension of Loxopholis ferreirai 237

Table 2. General information of previous and new occurrence observations of Loxopholis ferreirai at Rio Negro and Rio Jaú, state of Amazonas, Brazil. SL = Sampling locality number; N = number of specimens analysed per locality. m a.s.l. = meters above sea level. Coordinates are given in Datum WGS84. * GPS coordinates taken from field of notebook Miguel Trefaut Rodrigues (MTR) due Tableto geographic 2. General information inconsistence of previous or andabsence new occurrence in Rodrigues observations & ofÁvila-Pires Loxopholis ferreirai 2005. at Rio Negro and Rio Jaú, state of Amazonas, Brazil. SL = Sampling locality number; N = number of specimens analysed per locality. m a.s.l. = meters above sea level. Coordinates are given in Datum WGS84.

SL Year Voucher number N Location Locality Municipality Latitude Longitude Elevation Reference (m a.s.l.) 1* 1995 MZUSP 57312 (holotype) 1 Island Lago do Prato, Arquilélago Novo Airão -2.7175 -60.7529 30 Rodrigues and de Anvilhanas Avila-Pires (2005) 2* 1995 INPA 283 (paratype) 1 Island Ilha do Açaí, Arquipélago Novo Airão -2.645 -60.7639 20 Rodrigues and de Anavilhanas Avila-Pires (2005) 3 2017 MPEG 32848-32870 23 Island Parque Nacional do Jaú Novo Airão -1.9044 -61.7044 21 This study 4 2018 MZUSP 106723-106725 3 Island Ilha Comunidade Santa Barcelos -1.3929 -61.7951 21 This study Helena 5 2017 MZUSP 106718 1 Right bank Igarapé do Piloto Barcelos -0.8930 -62.9934 31 This study 6 2017 MZUSP 106713-106717 5 Island Ilha do Piloto Barcelos -0.8617 -62.9964 30 This study 7 2018 MZUSP 106732-106734 3 Island Ilha das Pombas Barcelos -0.8419 -62.9667 34 This study 8 2017 MZUSP 106719-106722 4 Island Ilha Paranã Uruá Barcelos -0.8288 -63.0077 34 This study 9 2018 MZUSP 106735-106736 2 Island Ilha Cacauali Santa Isabel do Rio Negro -0.4825 -64.8280 35 This study 10 2018 MZUSP 106730-106731 2 Island Poção Janatuba Santa Isabel do Rio Negro -0.3427 -65.4282 47 This study 11 2018 MZUSP 106726-106727 2 Island Ilha Uruari Santa Isabel do Rio Negro -0.3334 -65.4225 47 This study 12 2018 MZUSP 106729 1 Island Ilha Comunidade Boa Vista Santa Isabel do Rio Negro -0.3335 -65.3951 43 This study 13 2018 MZUSP 106728 1 Left bank Lago Mamiará Santa Isabel do Rio Negro -0.2945 -65.4078 53 This study

the first record of L. ferreirai outside the flooded fluvial During this trip (February 2017) the water level of Rio islands. As in Arquipélago de Anavilhanas, the islands Negro was rising, so that these islands were partially of Arquipélago de Mariuá are flooded in the wet season. flooded, and there were still portions of it above the water level. However, until now, L. ferreirai specimens were only found on the flooded portion of Rio Negro islands. We also found four specimens of L. ferreirai

in Ilha do Piloto, around 1700 h on the 16th of February 2017 (27.7o C air temperature [AT]; 86% of Relative Humidity [RH]). These four specimens were apparently resting, found inside rotten and broken tree trunks in decomposition on the flooded igapó. One of these specimens jumped on the water to escape, and minutes after was found again on the same tree trunk. Two specimens were found foraging on the igapó vegetation, one in dry twigs at 1711 h on the 17th of February 2017 (29.5o C AT; 80% RH) and one in a floating tree trunk at 1100 h on the 19th of February 2017. The remaining three specimens of Ilha do Piloto were found at night, Figure 1. Geographic distribution of Loxopholis ferreirai at as we revolved rotten trunks on the flooded igapó. The the flooded forests (igapó) of the Rio Negro and Rio Jaú. Data specimen from Ilha Anauali was active on a broken from holotype (1) and paratype (2) are represented by black and rotten tree trunk and led to the water during the triangles (Arquipélago de Anavilhanas). New records (3–13) capture attempt. The only specimen recorded outside are represented by black squares (fieldwork of 2017) and islands during this trip was found at 1640 h on the 19th black circles (fieldwork of 2018) and are detailed on Table 2. of February 2017 (30.0o C AT; 79% RH), foraging on The “*” symbol indicates sampling localities outside islands. the leaf litter of dry igapó forest, however distant few The cities of Novo Airão, São Gabriel da Cachoeira, Santa Isabel do Rio Negro and Barcelos are represented by white meters from the water line. Additionally, we found a circles, while Manaus, the capital of the state of Amazonas, is pair of L. ferreirai eggs (aprox. 0.7 cm each) deposited represented by a white circle with a black dot. See Table 2 for inside a rotten and broken tree trunk in decomposition, further geographic information. at ca. 1.5 m of the water level, on the flooded igapó 238 Sergio Marques-Souza et al.

At Parque Nacional do Jaú, PLVP and colleagues collected 23 specimens, all found inside a dry igapó forest. The site is seasonally flooded but at the time of sampling there were large portions of exposed land. All specimens were found among the leaf litter at the edge of the water (which is part of the flooded igapó). During the expedition along the Rio Negro, we found 49 specimens in seven localities distributed in the municipalities of Barcelos and Santa Isabel do Rio Negro. The large number of specimens obtained was due to a massive sampling effort of active search for longer periods of days. The specimens collected were found in the same type of habitats that we described above, mostly inside broken tree trunks in decomposition. Here, Figure 2. Eggs from Loxopholis ferreirai deposited on a rotten we selected one to three specimens to represent each and broken tree trunk, about 1.5 m high from the water level, locality. Specimens and localities are shown in Table on the flooded igapó forest (Ilha Anauali, municipality of 2 and Figure 1. All specimens were found in islands, Barcelos, state of Amazonas, Brazil). Ruler of 5 cm digitally except for one specimen collected on the left bank of inserted at the photography considering as reference the visible Rio Negro, Lago Mamiará (MZUSP 106728; Fig. 3). size of the TOB right hand index finger. Our results showed that L. ferreirai is not restricted to the islands of Arquipélago de Anavilhanas as previously known (Rodrigues and Avila-Pires, 2005), but occurs along other fluvial islands of Rio Negro and in the forest (Fig. 2). The eggs were collected and brought to igapó vegetation associated with its margins. Therefore, Universidade de São Paulo, maintained in the substract we suggest that L. ferreirai should be considered were they were collected (rotten wood), in a terrarium endemic to the entire range of flooded igapó forest of with room temperature (c.a. 25o C). The humidity was Rio Negro and Rio Jaú, a Rio Negro tributary. Overall, not controlled, but we sprayed water daily, in an attempt our extensive observations suggest that L. ferreirai does to recreate the local humidity conditions where the eggs not occur in the adjacent, unflooded terra-firme forests were collected. The eggs hatched after a month, when in Rio Negro basin. For instance, the terra-firme forests the species identification was confirmed. Unfortunately, adjacent to igapós are occupied by L. percarinatum, the newborns died shortly after they hatched, and a closely related species of L. ferreirai (Brunes et al., quickly rot, preventing them of being fixed. 2019), but the two species can occur in syntopy in the igapó vegetation. We also revealed that, similarly to other lizards occupying flooded environments (e.g. Kentropyx altamazonica Cope, 1875), L. ferreirai is scansorial, climbing the igapó vegetation to forage, oviposit and search for refuge, but also using water to escape from predators, remaining submerged for at least 45 seconds. Additionally, L. ferreirai uses the same microhabitats of its congeners, the non-flooded leaf litter. Our data suggest that L. ferreirai may present a broader heterogeneity of micro-habitat range than other Loxopholis species. However, more studies are needed to further verify this hypothesis, once our knowledge about Loxopholis natural history in general (as well as other leaf litter lizards) is anecdotal at best.

Figure 3. A male specimen of Loxopholis ferreirai (MZUSP Acknowledgements. We are grateful to Rafael de Fraga for 106728) from Lago Mamiará, municipality of Santa Isabel do providing a pre-peer review letter and helpful comments on an Rio Negro, state of Amazonas, Brazil. earlier version of this manuscript, Pedro Peloso, Pedro Pinna, Natural history notes and range extension of Loxopholis ferreirai 239 and Luisa Maria Diele-Viegas for their helpful comments and Underwood, W., Anthony, R., Cartner, S., Corey, D., Grandin, T., suggestions. We are deeply grateful to Pedro Peloso, Marcelo Greenacre, C.B., Gwaltney-Bran, S., McCrackin, M.A., Meyer, Sturaro and João Carlos Costa, for allowing us to use data collected R. and Miller, D. (2013): AVMA guidelines for the euthanasia by them for PARNA Jaú; Agustín Camacho, Camila Moreira, Ivan of animals: 2013 edition. Schaumburg, IL: American Veterinary Prates, José Cassimiro, José Mário Ghellere, and Renato Recoder Medical Association. for helping at the Rio Negro’s expedition. In Barcelos, we had Uzzel, T.M (1965): Teiid lizards of the genus Echinosaura. Copeia the kindly support of Valdeci’s family. We thank Aline Benetti 1965: 82–89. and Hussam Zaher for integrating our specimens into the MZUSP Uzzell T.M., Barry J.C. (1971): Leposoma percarinatum, a collection. Funding for this study was provided by Fundação de unisexual species related to L. guianense; and Leposoma ioanna, Amparo à Pesquisa do Estado de São Paulo (FAPESP), processes a new species from pacific coastal Colombia (Sauria, Teiidae). 2012/10163-1; 2017/08357-6 (SMS); 2016/03146-4 (TOB), and Postilla 154: 1–39. 2011/50146-6 (MTR). MTR also thanks Conselho Nacional de Vásquez-Restrepo, J.D., Ibáñez, R., Sanchez-Pacheco, S.J. & Desenvolvimento Científico e Tecnológico (CNPq), process Daza, J.M. (2019): Phylogeny, taxonomy, and distribution 301778/2015-9. We conducted this research in accordance with of the Neotropical lizards genus Echinosaura (Squamata: Brazilian legislation, governing standards of ethical procedures for Gymnophthalmidae), with the recognition of two new genera in collecting and scientific studies, and under consent and approval Cercosaurinae. Zoological Journal of Linnean Society, zlz124, of Instituto Chico Mendes de Conservação da Biodiversidade https://doi.org/10.1093/zoolinnean/zlz124 (ICMBio - permission 30309-11 and 28198-11). Vitt, L.J., Sartorius, S.S., Avila-Pires, T.C.S., Espósito, M.C.E. (2001): Life at the river´s edge: ecology of Kentropyx References altamazonica in Brazilian Amazonia. Canadian Journal of Zoology 79: 1855–1865. Avila-Pires, T.C.S. (1995): Lizards of Brazilian Amazonia (Reptilia: Vitt, L.J., Blackburn, D.G. (1991): Ecology and life history of the Squamata). Zoologische Verhandelingen 299: 1–706. viviparous lizard Mabuya bistriata (Scincidae) in the Brazilian Brunes, T.O., da Silva, A.J., Marques-Souza, S., Rodrigues, M.T., Amazon. Copeia 4: 916–927. Pellegrino, K.C. (2019): Not always young: The first vertebrate Wohl, E.E. (2007) Hidrology and Discharge. In: Large Rivers: ancient origin of true parthenogenesis found in an Amazon leaf Geomorphology and Management, p.29–44. Gupta, A., Ed., litter lizard with evidence of mitochondrial surfing on the wave England, Wiley. of a range expansion. Molecular Phylogenetics and Evolution 135: 105–122. Junk, W.J. (2000): Neotropical floodplains: a continental-wide view. In: The Central Amazon floodplain: actual use and options for sustainable management, p.5–24. Junk, W.J., Ohly, J., Piedade, M.T.F., Soares, M.G.M., Eds., Leiden, Netherlands, Backhuys Publishers. Marques-Souza, S., Pellegrino, K.C.M., Nunes, P.M.S., Junior, M.T., Gordo, M., Carvalho, V.T., Almeida, A., Oliveira, D.P., Frazão, L., Hrbek, T. Farias, I.P. & Rodrigues, M.T. (2015) On the discovery of bisexual populations of the parthenogenetic lizard Leposoma percarinatum (Gymnophthalmidae), with insights into the origin of parthenogenesis in Leposoma. South American Journal of Herpetology 10(2): 121–131. Marques-Souza, S., Prates, I., Fouquet, A., Camacho, A., Kok, P.J.R., Sales-Nunes, P.M., Dal Vechio, F., Recoder, R., Mejia, N., Teixeira-Junior, M., Barrio Amorós, C., Cassimiro, J., Lima, J.D., Sena, M.A. & Rodrigues, M.T. (2018): Reconquering the water: Evolution and systematics of South and Central American aquatic lizards (Gymnophthalmidae). Zoologica Scripta 47: 255–265 Pellegrino, K.M., Brunes, T.O., Souza, S.M., Laguna, M.M., Avila- Pires, T.C.S., Hoogmoed, M.S., Rodrigues, M.T. (2018): On the distinctiveness of Amapasaurus, its relationship with Loxopholis Cope 1869, and description of a new genus for L. guianensis and L. hoogmoedi (Gymnophthalmoidea/Ecpleopodini: Squamata). Zootaxa 4441: 332–346. Rodrigues, M.T., Avila-Pires, T.C.S. (2005): New lizards of the genus Leposoma (Squamata, Gymnophthalmidae) from the Accepted by Pedro Pinna lower Rio Negro, Amazonas, Brazil. Journal of Herpetology 39: 541–546.