Herpetology Notes, volume 13: 113-123 (2020) (published online on 11 February 2020)

Diet composition of bolivianus Gallardo, 1961 (Anura, ) of the Curiaú Environmental Protection Area in the Amazonas river estuary

Mayara F. M. Furtado1 and Carlos E. Costa-Campos1,*

Abstract. Information on a species’ diet is important to determine conditions and resources and to assess the effect of preys on the species distribution. The present study aimed describing the diet of Lysapsus bolivianus in the floodplain of the Curiaú Environmental Protection Area. Individuals of L. bolivianus were collected by visual search in the floodplain of the Curiaú River. A total of 60 specimens of L. bolivianus were euthanized with 2% lidocaine, weighed, and their stomachs were removed for diet analysis. A total of 3.020 prey items were recorded in the diet. The most representative preys were: Diptera (36.21%), Collembola (16.61%), and Hemiptera (8.31%), representing 73.20% of the total consumed prey. Based on the Index of Relative Importance for males, females, and juveniles, the most important items in the diet were Diptera and Collembola. The richness of preys recorded in the diet of L. bolivianus in the dry season was lower than that of the rainy season. Regarding prey abundance and richness, L. bolivianus can be considered a generalist species and a passive forager, with a diet dependent on the availability of preys in the environment.

Keywords. Amphibia; Eastern Amazon, Niche overlap, Predation, Prey diversity, Pseudinae

Introduction (Toft 1980; 1981; Donnelly, 1991; López et al., 2009; López et al., 2015). The diet of most anuran species is composed mainly of The genus Lysapsus Cope, 1862 is restricted to South arthropods and because of the opportunistic behaviour of America and comprises aquatic and semi-aquatic anurans many species, anurans are usually regarded as generalist inhabiting temporary or permanent ponds with large predators (Duellman and Trueb, 1994). Data on diet of quantities of aquatic vegetation (Prado and Uetanabaro, anurans can help to understand life history, identify 2000; Garda et al., 2007). The genus Lysapsus includes environmental conditions and consequences of habitat four species: L. bolivianus Gallardo, 1961, L. caraya alterations (e.g., different stages of deforestation), Gallardo, 1964, L. limellum Cope, 1862, and L. laevis prey species distribution, and reasons for population (Parker, 1935) (Aguiar-Jr et al., 2007). These species are fluctuations (Lips et al., 2005; López et al., 2015). distributed in the basins of Amazon River, - Anuran feeding mechanisms evolved in response to Araguaia, Paraná river, and the Rupununi savannahs, selective pressures and phylogenetic constraints, such respectively (Garda et al., 2010) that diets are influenced by morphology, physiology, and Lysapsus bolivianus Gallardo 1961 is a diurnal and behaviour (Toft, 1980). Prey availability of appropriate nocturnal species with snout-vent length (SVL) up to types and sizes is a factor that influences the diet of 20.2 mm in males and 23.5 mm in females, found in anurans, but few studies have assessed prey availability the water surface on loose clusters of emergent aquatic in the environment to allow interpretation of the results vegetation (Bosch et al., 1996). Males call from a horizontal position on leaves of aquatic macrophytes (Furtado et al., 2014). The species is listed as Data Deficient by the IUCN Red List of Threatened Species, and there is little information on its extent of occurrence, 1 Universidade Federal do Amapá, Departamento de Ciências status and ecological requirements (Angulo, 2008). Biológicas e da Saúde, Laboratório de Herpetologia. Rod. JK Km 2, Bairro Jardim Marco Zero, CEP 68902-280, Macapá, In this study we describe aspects of the diet of L. Amapá. bolivianus in a floodplain area of northern , eastern * Corresponding author. Email: [email protected] Amazonia in order to: (1) determine diet composition in 114 Mayara F. M. Furtado & Carlos E. Costa-Campos adult and juveniles; (2) assess possible differences in the numerical and volumetric percentages of each prey diet composition according to life stage and sex, and (3) category were determined for pooled stomachs. We determine if there is seasonal variation in diet. calculated an index of relative importance (IRI) for each prey taxon using the sum of the percentages of number Materials and Methods (N%), frequency (F%) and volume (V%): IRI = F (%) + N (%) + V (%) / 3 (Biavati et al., 2004). The study was conducted in a temporary pond located The niche breadth was calculated using the percentage in Toca da Raposa Farm (0.15019º N; 51.03847º W) of each prey item in the stomach, according to the and inserted in Curiaú Environmental Protection Area, formula: B = [(Σ P2 )-1 - 1] (n - 1)-1, where B = north of the municipality of Macapá. The climate of the a j ij a standardized trophic niche breadth; P = proportion of region is Equatorial (Am) according Köppen-Geiger ij food category j in the diet of species i; n = total number classification (Alvares et al., 2014) and the average of food categories. The standardized Levin’s index for temperature is 35.2 °C, varying seasonally between niche breadth was used (Krebs, 1999), which varies 27.6 to 41.0 °C. The dry season goes from August to from 0 (exclusive use of a single prey category) to 1 December (160 mm) and the rainy season begins in (equal use of all prey categories). January, reaching the highest rainfall in March and July, The food niche overlap between sexes was calculated with annual average of 1.530 mm (NHMET, 2012). using Pianka’s symmetric index (Pianka, 1973) as given We carried out our sampling by active visual search by the equation O = ∑ P P /√∑ P 2 ∑ P 2, where from October 2011 to September 2012. Three researchers jk ij * ik ij * ik j and k refer to the two sexes under comparison, O – walked in a straight line the five transects of 1 km each jk niche overlap, P – proportion of food component i. The (50 m apart from one another), in floating vegetation i index ranges from 0 to 1, with 1 indicating complete composed of Nymphoides indica (L.) Kuntze and overlap (Krebs, 1999). Salvinia auriculata Aubl (Crump and Scott Jr., 1994). We performed Statistical tests using BioEstat 5.3 We collected a total of 60 individuals of L. bolivianus, software (Ayres et al., 2007) at a significance level of p totalling 144 hours of sampling. ≤ 0.05. Before performing parametric tests, all variables In the laboratory, measurements of SVL were obtained were tested for normality and homoscedasticity of with a digital calliper (precision 0.01 mm). ’ sex variances. We tested for differences between males, was determined based on the presence of secondary females and juveniles in the number of prey and in sexual traits (vocal sac for males) or by direct observation the mean prey volume consumed per using one- of gonads. The females were considered adult when way analysis of variance (ANOVA). We compared the their ovaries were convolutes and enlarged. Individuals sizes (SVL) of both sexes by using the Kruskal-Wallis lacking these characteristics were considered juveniles. test. The niche breadth was analysed with the software For the diet analysis, collected specimens were Ecological Methodology (Krebs, 1999) and the niche euthanized with anaesthetic (2% lidocaine), fixed with overlap, with the software EcoSim 7.0 (Gotelli and 10% formalin, and preserved in 70% ethanol, according Entsminger, 2001). to Heyer et al. (1994). Data on diet were obtained through the analysis of stomach content of dissected Results specimens and prey items were identified to the Order level. We identified preys according with identification Diet composition.—We examined the stomach keys by Borror and Delong (2011) and Rafael et al. contents of 60 individuals of L. bolivianus (25 males, (2012). Anurans voucher specimens were deposited in 22 females, and 13 juveniles). Females (SVL 19.8 ± 1.4 the herpetological collection of Universidade Federal mm, range: 16.4–23.5 mm) were significantly larger do Amapá. than males (SVL 17.6 ± 1.1 mm, range: 14.1–20.8 mm; We measured length and width for all prey items Kruskal Wallis 12.0433; p < 0.0001). Juveniles varied in under stereomicroscopes with graduated oculars and SVL from 12.7 to 16.9 mm (15.5 ± 0.9 mm). their volumes were estimated using the equation for an We recorded a total of 3.020 items with a mean of 50.3 ellipsoid (cf. Vitt and Zani 2005): V = 4/3 π (length/2) preys per stomach. The most frequent prey items were (width/2)2. The frequency of items consumed (number Diptera (35.7%), Collembola (17.7%), and Hemiptera of stomachs containing the prey item), frequency of (8.2%), representing 60.8% of the total consumed prey occurrence (number of stomachs containing the prey (Table 1). These prey categories also presented higher category i, divided by the total number of stomachs) and IRI values (Table 1). Diet composition of Lysapsus bolivianus in the Amazonas river estuary 115

3.56 80.85

0.0016

0.06 0.68 0.82 22.45 9.15 103.05 0.07 0.19 81.15 0.71 2.22 113.24 0.008 0.09 22.25

------JUVENILES

1 11.11 1 11.11 2 22.22 3 8.11 0.011 0.03 32.44 6 16.22 9 28.13 5 2.18 0.015 0.01 49.78

47.60

collected in a temporary pond located in Toca da Raposa Farm and inserted in a FEMALES collected in a temporary pond located in Toca da Raposa Farm and inserted in a Curiaú Curiaú a in inserted and Farm Raposa da Toca in located pond temporary a in collected ------2 6.25 1 3.13 0.009 0.029 9.38 ------1 3.13 1 3.13 0.012 0.039 6.26 ------10 0.44 8 3.49 6.08 2.66 4.82 ------1 ------11.11 ------7 ------77.78 ------1 --- 2.70 --- 9 1 ------24.32 2.70 --- 2 --- 2.04 --- 5.41 --- 2 5.52 ------5.41 24 --- 7.25 64.86 --- 2.47 ------6.68 ------1 13.04 --- 3.13 ------1 3.13 ------71.61 ------223.79 ------1 --- 0.44 1 1 0.44 3.13 --- 0.0005 1 0.44 1 2.18 0.33 1 0.95 0.33 1.19 2.66 1.16 1.06 ------2 --- 6.25 2 6.25 --- 0.019 0.06 --- 12.52 ------27 84.38 ------109 ------1 11.11 ------1 2.70 1 2.70 0.35 0.95 5.72 ------Lysapsus bolivianus ------Lysapsus bolivianus Lysapsus

2.50

0.0003

0.47 0.59 72.70 0.310 0.388 42.63 0.047 0.036 36.94

MALES 9 11.25 ------12 15.00 15 11.54 ------

6 6 57.50 4 5.00 2 2.50 0.02 0.02 7.51 3 2.31 2 1.54 0.39 0.30 3.95 --- 4 5.00 2 0.03 0.020 0.025 5.03 1 1.25 1 1.25 0.0003 1 1.25 1 1.25 2.68 3.35 3.62 4 ------

Diptera Hemiptera ------Collembola Diptera Araneae Collembola Hemiptera Diptera Acari Decapoda (rainy) --- Collembola --- Diplura --- 25 Odonata --- 31.25 Hemiptera nymph --- Coleoptera larvae --- 2 1 2.50 1.25 --- 1 1 1.25 1.25 1.00 2.95 1.25 3.69 Acari 4.17 3.73 Araneae Collembola Odonata --- 33 25.38 --- Hemiptera nymph ------

Neuroptera larvae Neuroptera larvae Neuroptera larvae 1 1.25 Nematoda 1 1.25 NIA 1.18 --- 1.48 --- 2.99 ------

– Food items found in the stomach of 60 specimens (adults and juveniles) of Food items found in the stomach of 60 specimens (adults and juveniles) of of juveniles) and (adults specimens 60 of stomach the in found items Food SVL TAXON N N% F % V % IRI N N% F % V % IRI N N% F % V % IRI 12.0– 12.9 13.0– 13.9 14.0– 14.9 15.0– 15.9

Table Table 1 Curiaú Environmental Protection Area. SVL = snout-vent-length, N = number of items consumed, N% = numerical percentage of occurrence,items, V = volume of items consumed, V% numerical F percentage of items, IRI = of relativeindex importance = for each category of items in diet,the = NIA non-identifiedfrequency arthropods. of items consumed, F% frequency of Environmental Protection Area. SVL = snout-vent-length, N = number of items consumed, N% = numerical percentage of items, F = frequency of items consumed, F% frequency of occurrence, occurrence, of frequency F% consumed, items of frequency = F items, of percentage numerical = N% consumed, items of number = N snout-vent-length, = SVL arthropods. Area. non-identified = Protection NIA Environmental diet, the in items of category each for importance relative of index = IRI items, of percentage numerical V% consumed, items of volume = V Table 1. Table 116 Mayara F. M. Furtado & Carlos E. Costa-Campos 5.49

0.0007

0.37 0.41 79.26

------4.37 0.54 0.23 39.38 JUVENILES 14.29

2 2.20 0.03 0.03 13.20 10 13

59 64.84

6.69 ------

FEMALES 2 50.00 0.30 7.39 152.46 ------1 --- 0.44 1 --- 0.44 --- 0.09 --- 0.04 --- 2 0.89 0.87 --- 2 0.87 2 0.014 0.88 0.006 1 1.75 0.44 0.008 0.004 1.329 ------80 34.93 ------10 10.99 ------3 3.30 2 2.20 0.0007 ------14 ------6.11 ------9 1 3.93 --- 0.44 --- 3.44 1 --- 0.44 --- 6 1.50 0.25 2.62 10.55 --- 3 0.11 ------1.31 --- 0.91 --- 3.18 ------1.39 ------2 4.39 0.87 --- 1 --- 0.44 8.52 ------3.72 ------2.55 --- 1 --- 1.10 1 1.10 1.38 1.52 2.70

------4 100.00 9 12.00 3 4.00 0.017 0.023 16.01 ------

5.70 8.86 --- 7.51 --- 5 6.67 --- 3 --- 0.02 --- 0.0005 0.00068 ------

0.0002 0.0027 0.0001

0.17 0.13 84.66 0.30 0.09 73.45

MALES 0.32 0.88 0.28 1.36 ------3 3.30 3 3.30 7.26 7.98 9.25

22 16.92 26 8.23 0.012 0.004 38.29 34 10.76 29 7.02 0.002 0.001 43.10

62.66 36.08

3 0.73 4 0.97 0.42 0.10 1.73 1 1.33 1 1.33 0.07 0.09 2.70 ------8 6.15 3 2.31 0.013 0.010 8.46 --- 88 67.69 10 3.16 8 2.53 0.0005 21 5.08 10 2.42 0.0005 20 6.33 8 2.53 0.0085

Coleoptera Coleoptera larvae --- 2 --- 1.54 Hymenoptera --- 2 Diptera --- 1.54 --- Culicidae pupae --- 3.90 --- 3.00 1 ------4.08 0.77 --- Gstropoda --- 1 ------0.77 --- Acari 0.79 ------0.61 Collembola ------1.74 --- Odonata ------95 Orthoptera 30.06 --- Hemiptera 1 Coleoptera ------0.32 --- Coleoptera larvae --- 1 1 1 ------0.32 0.32 2 0.32 --- 4.55 1 0.63 Hymenoptera 1 0.32 1.44 --- 2 0.32 Diptera 0.88 1.11 0.63 0.35 3 --- 0.28 4.67 0.95 0.11 1.00 0.73 Acari 1.48 --- 0.67 --- 1.76 --- 198 Araneae ------Collembola ------Odonata nymph ------149 4 --- Orthoptera 2 --- 0.97 0.48 Hemiptera --- 2 --- 2 --- Hemiptera nymph 0.48 3 0.48 --- 13.65 --- 0.73 2.38 28 3.31 --- 3 6.78 0.58 --- 2.56 --- 11 0.73 1.16 ------2.66 6 0.66 --- 1 --- 0.12 6.59 7 --- 0.16 1.33 3 0.03 7.69 ------1.51 3.30 1 5 9.45 --- 2.10 5.49 --- 4 1.33 --- 2.31 2.94 5.33 10.66 --- 9.98 3.23 --- 2 14.26 13.31 ------2.67 7.10 --- 2.48 ------3.30 ------9.10 ------Neuroptera larvae 3 2.31 2 1.54 0.09 Nematoda 0.07 3.87 --- NIA ------Neuroptera larvae 1 0.32 1 0.32 NIA 0.04 0.01 0.64 ------

Continued. SVL TAXON N N% F % V % IRI N N% F % V % IRI N N% F % V % IRI 16.0– 16.9 17.0– 17.9

Table 1. Table Diet composition of Lysapsus bolivianus in the Amazonas river estuary 117 ------JUVENILES

------

2.22 ------4.30 ------4.7796 1.6652 6.4241

21.71 8.33 ------7.78148 0.05942 0.00005 4.91061

14.2401

FEMALES 61.75 18 9.84 0.31 0.17 71.642

6 3.28 2 1.09 1.43 0.78 4.63 ------55 73.33 9 12.00 2.53 3.38 27 86.458 14.75 8 4.37 0.047 0.026 19.13 ------15 3.99 37 9.84 3 12 0.80 18.4639 3.19 0.015 0.004 13.03 ------113

3.22 4 2.19 4 0.03 0.0030 0.00164

0.0001

MALES 0.48 5.68 1.38 2.15 ------0.27 8.43 2.26 1.56 3 1.64 1 0.55 5.38 2.94 3.17 ------

34 8.23 2.53 0.61 48.87 11 2.95 0.006 0.002 18.77 21 5.63 0.31 0.08 69.73

40.44 64.08

8 2.14 4 1.07 0.0004 1 0.27 1 1 0.27 0.27 7.83 1 2.10 0.27 1.24 3.79 --- 1.02 --- 0.87 --- 2 1.09 --- 1 --- 0.55 0.55 --- 0.30 --- 1.74 ------15 4.26 6 0.04 0.00023 ------6 3.28 2.00 1.09 0.011 0.006 4.37 ------

Coleoptera Coleoptera larvae 1 16 0.24 3.87 Hymenoptera 1 7 0.24 Diptera 1.69 5 0.08 4.33 Culicidae pupae 1.21 0.02 2.00 1.05 1 0.49 5.92 --- 0.24 167 Acari --- 1 ------Araneae 0.24 --- Collembola 1.15 --- 0.28 --- Orthoptera --- 0.58 10 59 Orthoptera nymph --- 2.68 --- 15.82 --- Hemiptera --- 5 1 1.34 --- 0.27Hemiptera nymph ------60.50 1 16.22 Coleoptera 16 ------9.43 0.27 --- 4.29 Coleoptera larvae 1 2.35 ------7 --- 1 --- 1.64 0.63 8 --- 1.88 1.23 --- 0.75 14.62 --- 2.14 Hymenoptera 1 1 --- 12.43 7 7 10.31 0.5464 1.23 Tricoptera 2.19 1.88 0.10 2 --- Diptera 0.39 0.54 7 0.13 --- 1.00 Culicidae pupae 0.10 2.51 ------3.83 4.06 1 Characiformes ------5 2.38 4 0.55 1.34 239 ------2.19 4 1.30 --- 1 --- 1.07 4 6.44 --- 0.55 --- Acari 1.22 --- 2.19 --- 0.17 0.33 Araneae ------3.67 2.52 ------0.09 Collembola --- 1 2.01 1.12 0.55 --- 1 5.04 --- 8 0.55 8 1 ------9.88 9.88 --- 1 --- 1 0.55 3 2 --- 0.55 0.55 3.70 0.20 2.47 11.45 1.00 0.010 0.12 14.13 0.11 0.55 0.013 18.30 12.35 39.7232 0.07 1.13 --- 1.12 ------Neuroptera larvae 3 0.73 1 0.24 0.08 Nematoda 0.02 0.97 --- 9 --- 2.18 --- 8 1.94 --- 0.007 0.002 --- 4.12 ------Neuroptera larvae ------Nematoda ------NIA 2 0.55 6 1.61 1 5 0.55 1.34 0.011 0.13 0.003 2.95 0.07 3 1.12 1.10 --- 1 ------0.55 0.10873

Continued. SVL TAXON N N% F % V % IRI N N% F % V % IRI N N% F % V % IRI

18.0– 18.9 19.0– 19.9 Table 1. Table 118 Mayara F. M. Furtado & Carlos E. Costa-Campos ------JUVENILES

------

1.5957 1.49 ------10.09 ------

0.04187 0.00007

35.30 22.201 19.98 ------

FEMALES 2.93 0.008 0.002 10.37 ------

63.03 39 10.37 4.83 1.29 73.833 54.51 22 8.27 0.40 0.15 62.832

15 3.99 8 2.13 1.17 0.31 6.22 ------237 4 1.48 28 10.53 2 6 0.01 2.26 0.00023 4.66 31 1.75 11.65 10 13.366 3.76 20.32 7.64 17.96 145 ------

11.76

0.0006

4.83 5.97 81.00 2.67 7.85 18.32 0.17 0.51 88.41

MALES 2.94 0.04 0.13 5.93 4 1.50 2 0.75 14.09 5.30 4.02 ------

17 20.99 13 38.24

2 5.88 2 5.88 0.0002 ------16 10.06 4 0.03 0.0010 0.00006 ------4 1.06 3 0.80 0.28 0.07 1.89 ------1 0.38 1 0.38 0.03 0.01 0.76 ------28 7.45 11.00

Odonata Orthoptera Hemiptera --- Hemiptera nymph 2 --- 2.47Psicoptera nymph --- 5 1 2 Coleoptera 6.17 --- 1.23 2.47 1 3 2.33 --- 3.70 1.23 1 Hymenoptera 2.87 3.43 0.11 --- 5.90 1.23 Diptera 4.24 0.13 --- 1 4 --- 11.29 --- 2.51 1.60 1.23 --- 2 --- 2.08 --- 4 0.53 --- 2.57 47 Acari 58.02 --- 1.06 1 --- 3.33 Araneae 6 --- 3.91 0.27 --- Collembola 1.60 1.04 3.12 --- 5 Odonata nymph --- 3.01 2 0.83 ------Orthoptera 1.33 4 ------5.88 1.07 3.49 1.06 1 Hemiptera ------1 3.13 Hemiptera nymph ------0.93 --- 14.39 ------Coleoptera ------9 42.31 0.27 --- 3.24 --- 23.11 1.00 7.06 ------5 ------Hymenoptera 1.88 --- 14.71 ------Diptera 1.96 1 ------1 39 Culicidae pupae 14.44 --- 2.94 --- 0.38 16 1.00 --- 1 --- Siluriformes --- 1 --- 2.94 6.02 2 17 Acari 1 50.00 --- 0.040 0.38 0.75 --- Araneae 2.94 --- 0.015 72.49 2 0.04 --- Collembola --- 20.46 27.25 1.17 0.75 2 ------9.84 6.27 0.75 2.97 ------4 ------1 1.12 1.50 ------1 0.38 --- 1.88 ------4.22 ------0.38 --- 1 --- 1.58 0.30 ------0.38 --- 1.66 0.11 1 ------1.92 0.38 15 --- 18 9.43 --- 11.32 --- 13.51 9 5 --- 5.08 --- 5.66 3.1447 2.44 0.011 --- 0.007 --- 16.98 ------Neuroptera larvae ------Nematoda --- NIA ------6 --- 1.06 --- 2 --- 0.53 --- 0.08 6 0.02 1.32 Neuroptera larvae 1.60 --- 1 ------0.27 ------0.15742 ------4 0.75 1 0.38 0.22 0.08 1.16 ------

Continued. SVL TAXON N N% F % V % IRI N N% F % V % IRI N N% F % V % IRI

20.0– 20.9 21.0– 21.9 Table 1. Table Diet composition of Lysapsus bolivianus in the Amazonas river estuary 119 ------JUVENILES

------

FEMALES MALES ------1 ------1.92 --- 1 ------1.92 1 2 ------3.85 26 1 1.92 --- 50.00 1 2.58 --- 1.92 --- 9 2.07 1.92 1 --- 4.96 17.31 25.00 4.45 3 5.50 3.98 0.33 1 75.00 --- 5.17 8.55 0.64 --- 25.00 3 --- 67.521 8.62 --- 1.73 75.00 ------2.73 43.27 --- 64.42 --- 68.13 --- 172.71 ------1 1.92 1.00 1.92 0.331 0.637 4.0583 ------2 1.26 1 0.63 0.23 0.15 1.94 ------6 3.77 3.00 1.89 0.017 0.011 5.66 ------

Odonata Odonata nymph --- Orthoptera ------Hemiptera ------Hemiptera nymph ------Coleoptera ------Hymenoptera ------Diptera ------1 --- Culicidae pupae --- 2 0.63 ------1.26 --- 1 ------2 ------Collembola 1 ------0.63 --- 3.77 15 Orthoptera --- 0.63 --- 9.43 41.04 2 ------Hemiptera ------4.74 25.81 6 --- 1.26 3 Coleoptera --- 9.86 --- 2.98 ------3.77 1.89 1.90 Diptera ------1 --- 1.72 2.88 3 --- 1.19 --- 0.63 ------1.08 1 --- 1.89 1 5.43 Hemiptera ------3.77 71 0.40 13.57 ------Coleoptera 44.65 0.63 1 ------11 0.25 0.01 ------0.63 19 6.92 --- 3.86 0.01 36.54 0.22 0.58 7 --- 1.26 0.14 --- 0.37 13.46 ------0.224 4.45 51.694 --- 0.431 ------50.14 ------Neuroptera larvae ------NIA ------6 3.14 2 NIA 1.26 0.11 0.07 4.43 ------

Continued. SVL TAXON N N% F % V % IRI N N% F % V % IRI N N% F % V % IRI 22.0– 22.9 23.0– 23.9

Table 1. Table 120 Mayara F. M. Furtado & Carlos E. Costa-Campos

The most frequently observed categories in the diet The diet niche breadth of juveniles (Ba = 2.74) was of adult males were Collembola (24.9%), followed wider than those of adult males (Ba = 2.42) and females by Diptera (19.1%) and Hemiptera (4.0%). The most (Ba = 2.48). The observed average of simulations of frequently observed categories in the diet of adult diet overlap of males, females, and juveniles was females were Diptera (57.8%). For juveniles, the most significantly higher than that expected by the null model frequent food items were Diptera (48.4%), followed (observed average = 0.96, simulated average = 0.58; p by Collembola (32.2%) and Coleoptera (5.2%) (Figure = 0.01). 1). There was no difference in the mean number of Seasonal variation in the diet.—During the dry prey items consumed by males, females and juveniles season the prey items richness recorded in the diet of L. of L. bolivianus (ANOVA, F = 0.3282; p = 0.726). bolivianus was lower than that of the rainy season (Figure The mean volume of prey ingested no differed among 2). Collembola (Heteromurus sp. and Pseudobourletiella males, females and juveniles (ANOVA, F = 0.0838; p sp.), Hemiptera (Belostomatidae), Coleoptera = 0.0102). (Mycetophagidae and Scarabaeidae), Tricoptera, Diptera The diet overlap was high between males and females. (Fanniidae), Diplura, Orthoptera larvae and Siluriformes were found only in individuals collected in the rainy season. Decapoda, Auchenorrhyncha, Coleoptera (Coccinellidae and Staphylinidae), Hemiptera larvae, Hymenoptera, Psicoptera larvae, Diptera (Culicidae), Characiformes, and Gastropoda were only recorded in individuals collected in the dry season. There was a high diet overlap between dry and rainy season (0.916).

Discussion Lysapsus bolivianus presents a generalist diet that consists mostly of arthropods (96.4%). Traditionally, small arthropods have been reported in many studies on food habits of anurans and particularly of Pseudinae frogs (e.g. Toft, 1980; Duré and Kehr, 2001; Teixeira et al., 2004). The selection of preys may be influenced by the type of habitat as well as their availability in the environment throughout the year (De-Carvalho et al.,

Figure 1. Frequency of occurrence (F%) of the food items Figure 2. Rainfall (mm), in bars, and prey items richness of the consumed by Lysapsus bolivianus collected in the temporary food items consumed by Lysapsus bolivianus, in lines, from pond located in Toca da Raposa Farm and inserted in a Curiaú October 2011 to August 2012 in the temporary pond located in Environmental Protection Area. (A) Males, (B) Females, and Toca da Raposa Farm and inserted in a Curiaú Environmental (C) Juveniles. Protection Area. Diet composition of Lysapsus bolivianus in the Amazonas river estuary 121

2008; Cuello et al., 2009; López et al., 2015). These The presence of Macrobrachium amazonicum factors associated with the differences in the occupation (Decapoda) in the stomach contents of L. bolivianus was of the habitat influence the types of preys ingested the first record of consumption of crustaceans in the diet by anurans (Van Sluys and Rocha, 1998). In general, of this species (Furtado et al. 2013). The consumption of the diet of most adult anurans mainly consists in decapod crustaceans has been recorded for P. paradoxa arthropods (Vitt and Caldwell, 2009), but because they (Downie et al., 2010), and isopod crustaceans for P. consume different invertebrates and its opportunistic bolbodactyla (Teixeira et al., 2004). behaviour, they are usually considered generalist The richness of preys found in the diet of L. bolivianus predators (Duelmann and Trueb, 1994). In our results, was higher in the rainy season, which agrees with those L. bolivianus is characterized as passive foragers, with a of Vaz-Silva et al. (2005) for L. laevis. The categories of generalist diet that uses floating vegetation as substrate preys found in the diet of L. bolivianus revealed different for foraging. peaks during the year, indicating that consumed items The orders Diptera, Coleoptera and Hemiptera were may vary in response to availability of preys in the the most consumed food items by L. limellum in environment (Toft, 1980). populations inhabiting southern latitudes (Province of Diet overlap among males, females, and juveniles Corrientes, Argentina: Duré and Kehr, 2001). Garda et was high, which could be explained by the similarity of al. (2007) studied aspects of the ecology of L. limellum food items recorded in the diet and by availability in the in the states of Amazonas and Pará and the main items environment. According to Gotelli and Graves (1996), a consumed were Diptera (34.85%), Hemiptera (22.73%) high level of food or unlimited resources sharing among and Odonata (9.09%). The main items found in the diet males, females, and juveniles of L. bolivianus suggests of L. laevis were Diptera (46.16%), Hemiptera (14.29%) resource partitioning influenced by biotic interactions among species, such as competition. and Coleoptera (8.79%) (Vaz-Silva et al., 2005). The inverse of the Simpson’s diversity index, used According to observations in the field, dipterans to measure niche breadth, considers prey categories are also very abundant in the studied area, and can be and the proportion of individuals consumed, therefore found in large concentrations in the emergent aquatic comprising the diversity and equitability of consumed vegetation, which is used by L. bolivianus for foraging, preys (Simpson, 1949). Thus, juveniles were expected protection from predators, and calling (E. Campos to have a wider niche breadth, since the distribution of pers. comm.). Similar information was reported for preys was evener among food items in juveniles of L. other pseudid species (Duré and Kehr, 2001; Teixeira bolivianus. et al., 2004; Miranda et al., 2006; Garda et al., 2007; Dietary studies are needed to determine similarities in Macale et al., 2008; Downie et al., 2010). These records pattern of diets of pseudid species. We also highlight support the data obtained in the present study, as the the importance of dietary studies that include prey order Diptera was the most frequent and numerically availability to better interpret the degree to which the most important prey category recorded in the diet similarities in the diets are due to historical factors and/ of L. bolivianus. The importance value indices of this or to prey availability in the environment. category for L. bolivianus support those by Teixeira et al. (2004) and Garda et al. (2007), which recorded Acknowledgments. We thank Ulisses Caramaschi for depositing higher importance value indices for Diptera in the Lysapsus bolivianus in the Herpetological Collection of the diet of bolbodactyla and , National Museum of Rio de Janeiro. Raimundo Nonato Picanço respectively. Souto (UNIFAP) and Bruno Bellini (UFRN) for their invaluable The presence of vertebrates in the diet of anurans was help in identifying arthropods. Marcelo Menin for helpful comments on the manuscript. Mateus Ramos for permission reported to large species of Leptodactylus, including where this study was conducted (Toca da Raposa Farm) and L. labyrinthicus (França et al., 2004), L. ocellatus SISBIO/ICMBio for the license granted, # 34238-1. (Teixeira and Vrcibradic, 2003; Sanabria et al., 2005), L. pentadactylus (Castro et al., 2011; Couto and Menin, References 2014), L. macrosternum (Sousa et al. 2016), Pseudis Aguiar, O. Jr., Bacci Jr. M., Lima, A.P., Rossa-Feres, D.C., Haddad, cardosoi (Miranda et al., 2006) and for P. Tocantins C.FB., Recco-Pimentel, S.M. (2007): Phylogenetic relationships (Neves et al. 2014). In the present study, the presence of Pseudis and Lysapsus (Anura, Hylidae, ) inferred of actinopterygian fishes indicates an opportunistic from mitochondrial and nuclear gene sequences. Cladistics 23: feeding behaviour of Lysapsus bolivianus. 455–463. 122 Mayara F. M. Furtado & Carlos E. Costa-Campos

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Accepted by Gonçalo Rosa