Acta Herpetologica 12(2): 205-208, 2017 ISSN 1827-9635 (print) © Firenze University Press ISSN 1827-9643 (online) www.fupress.com/ah DOI: 10.13128/Acta_Herpetol-20547 Diet of juveniles of the venomous frog Aparasphenodon brunoi (Amphibia: Hylidae) in southeastern Brazil Rogério L. Teixeira1, Ricardo Lourenço-de-Moraes2, Débora C. Medeiros3, Charles Duca3, Rogério C. Britto4, Luiz C.P. Bissoli5, Rodrigo B. Ferreira3,* 1 In memoriam 2 Laboratório de Herpetologia e Comportamento Animal, Departamento de Ecologia, Universidade Federal de Goiás, Campus Samam- baia, 74001-970 Goiânia, GO, Brazil 3 Laboratório de Ecologia de Populações e Conservação, Programa de Pós-graduação em Ecologia de Ecossistemas, Universidade Vila Velha. Rua Comissário José Dantas de Melo, Boa Vista II, 29102-920 Vila Velha, ES, Brazil. *Corresponding author. E-mail: rodrigo- ecologia@yahoo.com.br 4 Associação Capixaba em Defesa da Água e da Mata Atlântica (ACADAMA). Rua Lourenço Roldi,  29665-000 São Roque do Canaã, ES, Brazil 5 Ello Ambiental. Av.Getúlio Vargas, 29700-010 Colatina, ES, Brazil Submitted on: 2017, 17th April; revised on: 2017, 29th May; accepted on: 2017, 27st September Editor: Daniele Pellitteri-Rosa Abstract. Seventy juvenile individuals of Aparasphenodon brunoi were collected on the low parts of tree trunks in an Atlantic Forest remnant. Arthropods were the dominant prey found in their stomachs. Coleoptera (adult and larvae) was the most important prey regarding prey frequency, number, weight, and index of relative importance. Secondary preys included Hymenoptera that was important regarding number of prey and Hemiptera that was important regard- ing prey weight. Trophic ontogeny was detected. The diversity of prey suggests A. brunoi is an opportunistic sit-and- wait predator. Keywords. Anura, ecology, Hylidae, predation, ontogeny, feeding pattern. Studies regarding dietary aspects of animals may provide important data for a better understanding of the fundamental niche, position in the food webs, feeding time strategy, and metabolic needs of a species (Chris- tian et al., 2007; Miller et al., 2010). Anurans are usually considered opportunistic predators regarding their feed- ing habits and most of them are arthropod consumers (Wells, 2007). Generally, the diet of anurans is related to their snout-vent length, ability to detect and capture prey as well as prey availability in the environment (Giaretta et al., 1998; Ferreira et al., 2015). Aparasphenodon brunoi Miranda-Ribeiro, 1920 is a hylid treefrog that uses bromeliads and hollows of tree trunks as diurnal refuge (i.e., bromelicolous species, sensu Peixoto, 1995) and reproduce in temporary ponds (Wogel et al., 2006). It is endemic to the coastal plain of Bra- zil (i.e., restinga habitat) from São Paulo to Bahia states (Ruas et al., 2013; Frost, 2016). This species has a highly toxic secretion that may be injected by bony spines on the head (Jared et al., 2015). Furthermore, some studies have shown that the toxicity of a frog is determined by the compounds sequestered from the diet during early devel- opment stage (e.g., Daly et al., 1994; Sime et al., 2000). There is little information on the ecology of juveniles of the venomous treefrog A. brunoi, and diet studies on adults have shown that this treefrog is a generalist forager of arthropods (Teixeira et al., 2002; Mesquita et al., 2004). The aim of this study is to describe the diet of juveniles 206 Rogério L. Teixeira et alii of A. brunoi in an Atlantic Forest remnant from south- eastern Brazil. The study area is in the coastal plain of Pontal do Ipiranga district, municipality of Linhares, Espírito Santo state, Brazil (19°13’S, 39°43’W). The region is character- ized by pastures and remnants of Atlantic Forest. The treefrogs were manually captured at night between Sep- tember and October 2000. According to Mesquita et al. (2004), individuals of this species smaller than 48 mm are considered juveniles. The individuals were anesthetized using lidocaine cream (5%) and fixed in a solution of for- malin 10% for 72 hours and then washed and placed in a solution of alcohol 70%. In laboratory the specimens were measured to the snout-vent length (SVL in mm), weighted (0.01 g precision) and dissected to determine the sex and to analyze the stomach content. The stomach contents were removed, spread in Petri dish and identi- fied to the lowest taxonomic level possible. Preys were dried on paper towels and the following parameters for each prey category were evaluated: occur- rence frequency, number, wet weight (0.0001 g precision), and index of relative importance (IRI). The IRI was cal- culated according to Pinkas et al. (1971) and the percent IRI (%IRI) according to Cortés (1997). For intraspecific comparison, IRI was calculated for adults of A. brunoi using data available on Teixeira et al. (2002). Intact preys were measured to the total length with a caliper (0.1 mm precision). The voucher specimens were deposited at the zoological collection of the Instituto Nacional da Mata Atlântica/Museu de Biologia Professor Mello Leitão (MBML), Espírito Santo State, Brazil. The data were tested for normality using Kolmog- orov-Smirnov test and for homogeneity using Bart- lett test. SVL and weight were compared between sexes using the analysis of variance (ANOVA). In this analysis, sex was the independent variable while SVL and weight were the dependent variables (Neter et al., 1990). Cluster analysis based on Euclidean distance was used to evalu- ate trophic ontogeny based on size classes of A. brunoi. Only the weight value of the main prey was used in this analysis and the data were log transformed. The relation- ship between prey length and anuran SVL was tested by a regression analysis. Mean and standard deviation are pro- vided. The significance level was set to 0.05. Seventy juvenile individuals of A. brunoi were col- lected on the low parts of tree trunks (< 1m). Adults were on trees canopy and were not collected. Thirty-two indi- viduals were juvenile males and 38 were juvenile females. Juvenile males ranged on SVL from 31.0 to 39.2 mm (mean = 34.3, SD = 2.0 mm) and in weight from 2.0 to 4.4 g (mean = 2.9, SD = 2.0 g). Juvenile females ranged on SVL from 32.1 to 43.6 mm (mean = 36.7, SD = 2.7 mm) and in weight from 2.1 to 6.7 g (mean = 3.6, SD = 2.7 g). The SVL and weight differed significantly between Table 1. Prey items of juveniles of Aparasphenodon brunoi from Espírito Santo state, southeastern Brazil. F = occurrence frequency; N = number of prey; W = weight; IRI = index of relative importance. Prey F %F N %N W %W IRI %IRI Insecta Blattodea 2 3.77 2 3.23 120.1 3.94 27.03 0.77 Coleoptera (Adult) 19 35.85 21 33.87 963.5 31.64 2348.53 67.26 Coleoptera (Larvae) 8 15.09 8 12.90 267.9 8.80 327.45 9.38 Hemiptera 3 5.66 3 4.84 946.5 31.09 203.36 5.82 Homoptera 1 1.89 1 1.61 1.8 0.06 3.16 0.09 Hymenoptera 7 13.21 10 16.13 127.1 4.17 268.16 7.68 Isoptera 1 1.89 1 1.61 19.3 0.63 4.23 0.12 Orthoptera 5 9.43 5 8.06 333.1 10.94 179.17 5.13 Insect remnant 3 5.66 - - 23.3 0.76 Arachnida Araneae 3 5.66 3 4.84 160.6 5.27 57.22 1.64 Crustacea Ostracoda 3 5.66 8 12.90 1.5 0.05 73.30 2.10 Other Anuran skin 5 9.43 - - 80 2.63 Total - - 62 100.00 3044.7 100.00 207Diet of juveniles of Aparasphenodon brunoi the sexes (F1,68 = 17.3, P < 0.01; F1,68 = 12.6, P < 0.01, respectively). Females were on average larger and heavier than males. Fifty-three (76%) individuals had food items in their stomach, representing 12 prey categories, most- ly arthropods, (Table 1). Coleoptera (adult and larva) was the most important prey regarding prey frequency, number, weight, and IRI. Secondary preys included Hymenoptera that was important regarding number of prey and Hemiptera that was important regarding prey weight. It was detected trophic ontogeny in A. bru- noi (Fig. 1). Specimens less than 36.9 mm had similar diet, possibly because they predated more Coleoptera (Fig. 2). Specimens larger than 36.9 mm had simi- lar diet by feeding mostly upon Hemiptera. The length of intact prey ingested by A. brunoi varied from 2.8 to 15.9 mm (mean = 8.9, SD 3.7). There was no relation- ship between total length of prey and anuran SVL (R² = 0.003, P > 0.05). The variety of prey categories suggests that A. brunoi is an opportunistic sit-and-wait predator. The diet of A. brunoi from another two populations had also a variety of prey categories (Teixeira et al., 2002; Mesquita et al., 2004). In fact, the feeding upon small arthropods has been reported in several studies of anurans (Teixeira and Coutinho, 2002; Ferreira and Teixeira, 2009; Ferreira et al., 2012). The presence of ostracods, common on tank bromeliads (Oliveira et al., 1994), indicates bromelicolous habit of A. brunoi at the studied fragment. Apparently, juveniles of A. brunoi feed upon different prey categories compared to adults. In our study, Coleop- tera was the most important prey (%IRI = 67.26). In Teixeira et al. (2002), Hymenoptera was the most impor- tant prey (%IRI = 20.1) for adults of A. brunoi. Coleop- tera was the least important prey (%IRI = 0.7) for this population of adults. In our study, A. brunoi feeds on different prey categories according to their SVL. Surpris- ingly, A. brunoi SVL was not related to prey size. Changes on diet correlated to anuran SVL have been reported for other species (Giaretta et al., 1998; Teixeira and Vrci- bradic, 2003; Ferreira et al., 2007). 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