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Diet and Clutch Size of Rhinella Castaneotica (Anura: Bufonidae) from a Forest Area in Serra

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Diet and Clutch Size of Rhinella Castaneotica (Anura: Bufonidae) from a Forest Area in Serra Use the following type of citation: North-western Journal of Zoology 2021: e211508 Paper Submitted to The North-Western Journal of Zoology 1 *Handling editor: Yurii Kornilev 2 *Manuscript Domain: Herpetology of the Americas 3 *Manuscript code: NwJZ_20_HA_12 4 *Submission date: 04_06_2020 5 *Revised: 15_03_2021 6 *Accepted: 23_03_2021 7 *No. of words (without abstract, acknowledgement, references, tables, captions): 2340 8 (papers under 700 words are not accepted) 9 *Editors only: 10 11 Zoology 12 Title of the paper: of 13 Diet and clutch size of Rhinella castaneotica (Anura: Bufonidae) from a forest area in Serra 14 do Navio, Amapá, Brazil Journal 15 Running head: 16 Diet and clutch size of Rhinella castaneotica paper 17 Authors (First LAST - without institution name!): 18 Vinícius Figueiredo, Edgar Santos, Fillipe Santos, Patrick Sanches, Carlos Campos North-Western 19 accepted 20 Key Words (at least five keywords): 21 Prey selection, trophic niche, niche breadth, reproduction, Brazil. 22 23 No. of Tables: 1 24 No. of Figures: 0 25 No. of Files (landscape tables should be in separate file): 1 26 27 Use the following type of citation: North-western Journal of Zoology 2021: e211508 nwjz-2 28 Diet and clutch size of Rhinella castaneotica (Anura: Bufonidae) from a forest area in 29 Serra do Navio, Amapá, Brazil 30 Vinícius A. M. B. de FIGUEIREDO*, Edgar Matos dos SANTOS, Fillipe PEDROSO- 31 SANTOS, Patrick R. SANCHES and Carlos Eduardo COSTA-CAMPOS 32 Universidade Federal do Amapá, Departamento de Ciências Biológicas e da Saúde, 33 Laboratório de Herpetologia, Campus Marco Zero do Equador, 68.903-419, Macapá, AP, 34 Brazil 35 * Corresponding author, V. A. M. B. de Figueiredo, E-mail: [email protected] 36 37 Abstract. We studied the diet and the clutch size of Rhinella castaneotica from a primary Zoology 38 forest in northern Brazil, municipality of Serra do Navio,of Amapá state. A total of 100 toads 39 (77 males and 23 females) were collected between April 2017 and March 2018 through active 40 searches. Thirty-four individuals (34%) had a total of 870 prey items in their stomach Journal 41 contents. Hymenoptera and Coleoptera were the most important and numerous prey items in 42 the diets of males and females, respectively. The niche breadth based on prey numbers was 43 0.24. Based on 9 females with eggs, thepaper number of ovarian eggs ranged from 199 to 720 eggs. 44 We conclude that R. castaneotica has a specialized diet and its clutch size is independent of 45 the female SVL andNorth-Western body mass. We highlight the importance of further studies that contribute accepted 46 to a better understanding of the conservation status and ecological aspects of such poorly 47 known anurans in the Amazon. 48 49 Key Words: prey selection, trophic niche, niche breadth, reproduction, Brazil. 50 Running title: Diet and clutch size of Rhinella castaneotica 51 52 Introduction Use the following type of citation: North-western Journal of Zoology 2021: e211508 nwjz-3 53 The natural history of a given species is characterized by its distinct life-history 54 components (Putnam 1994). Diet, reproduction and micro-habitat use make up the 55 components most often examined in the majority of studies regarding the natural history of 56 anurans species (De-Carvalho et al. 2008, Freitas et al. 2008, Luria-Manzano & Gutierrez- 57 Mayen 2014, Jorge et al. 2015, Pinto & Menin 2017). 58 As pointed by Toft (1981), several factors influence the diet of anurans, such as the 59 size of prey, foraging strategies, body size, and physiological constraints. Furthermore, the 60 diet may vary ontogenetically and due to sexual differences, as males and females have 61 different energy needs (Lima 1998, Wu et al. 2005). As result, there is plasticity involving 62 anurans within trophic chains, which is expressed by their acting as predators and prey Zoology 63 (Toledo et al. 2007). of 64 The genus Rhinella is currently composed of 92 species recognized, distributed from 65 southern Texas, USA to southern South America (Frost 2020). Species often show sexual Journal 66 dimorphism, as recorded in several species such as R. achalensis, R. marina, R. rubescens, 67 and R. diptycha (Monnet & Cherry 2002, Jofré et al. 2005, Arantes et al. 2015). In general, 68 the toads of the genus Rhinella exhibitpaper explosive breeding events (Machado & Bernarde 69 2011), and these reproductive behaviors have been well-documented (Pombal-Junior & 70 Haddad 2005, Vargas-SalinasNorth-Western 2007, Machado & Bernarde 2011, Bowcock et al. 2013, accepted 71 Valentim et al. 2013, Costa-Campos et al. 2016). 72 The R. margaritifera species group currently contains 14 species, including the poorly 73 known R. castaneotica (Caldwell 1991). This small-sized species (snout-vent length 18.4– 74 23.6 mm in males and 18.9–26.3 mm in females) is present in the Amazon basin of Bolivia, 75 Brazil, Colombia, and Peru (IUCN 2015). Rhinella castaneotica are nocturnal and terrestrial 76 toads; south of the Amazon River males call sporadically throughout the rainy season in Use the following type of citation: North-western Journal of Zoology 2021: e211508 nwjz-4 77 ponds and small holes filled with water or from within empty Brazil nut capsules called 78 “castanheiras” (Caldwell 1993, Lehtinen et al. 2004). 79 Herein, we provide data on some aspects of the natural history of R. castaneotica, including 80 information on the diet composition and reproduction in a population from a forest area in 81 Amapá State, Brazilian Amazonia. 82 83 Material and Methods 84 We conducted surveys at the Cancão Municipal Natural Park (0°54’39.04”N, 85 52°0’29.72”W), Serra do Navio municipality, Amapá state, Brazil. The region is composed of 86 a mosaic of seasonal ombrophilous dense forest and floodplain (igapó) forest (Drummond et Zoology 87 al. 2008). The climate is Am (Köppen-Geiger classification;of tropical monsoon, without a dry 88 season), and the mean annual rainfall is 1,885 mm and temperature 27 °C (Peel et al. 2007). 89 Individuals examined in this study were collected for a study of the natural history of Journal 90 anurans in the eastern Amazon (diet, reproduction and helminth parasites) using visual and 91 auditory active searches (Heyer et al. 1994) in temporary ponds and on the forest floor during 92 five consecutive nights monthly frompaper April 2017 to March 2018. Immediately after 93 collection, all individuals were weighed with a digital balance (0.1 g precision), killed using 94 lidocaine 2%, and theirNorth-Western snout-vent length (SVL) was measured with a digital caliper (0.01 accepted 95 mm precision). The sex was recorded for each individual based on direct observation of 96 gonads. 97 Individuals were fixed in 10% formaldehyde solution, preserved in 70% ethanol, and 98 housed in the Herpetological Collection of the Universidade Federal do Amapá, state of 99 Amapá, Brazil (vouchers CECC 1419, 1420, 1449, 1450, 1452–1463, 1480–1484, 1527– 100 1529, 1854–1866, 1872, 1873, 1875–1880, 1882–1888, 2020, 2063–2079, 2092–2099, 2126– 101 2135, 2142–2155, 2259–2262). Use the following type of citation: North-western Journal of Zoology 2021: e211508 nwjz-5 102 For dietary analysis we removed the stomach of each specimen through a ventral 103 vertical incision and subsequently we set all extracted stomach contents into plastic tubes 104 filled with 70% ethanol. Direct extraction of stomach rather than non-lethal techniques (e.g. 105 stomach-flushing) was justified because individuals from our study were also part of research 106 on helminth parasites, the methodology for which requires a complete gastrointestinal 107 analysis and euthanasia. 108 We identified the prey items to the lowest possible taxonomic category following 109 Rafael et al. (2012) and Baccaro et al. (2016). To avoid misclassifications of food items and 110 misestimates in prey sizes we included only intact items for the content analysis (e.g. wings 111 and fragmented exoskeleton parts were not considered in the sample). We measured the Zoology 112 maximum length and width of each prey item with the aidof of millimetric ocular lens to obtain 113 the volume through the Ellipsoid Volume Formula following Griffiths & Mylotte (1987), 114 where l is the prey length and w is the prey width: Journal 115 116 In addition, we calculated the Indexpaper of Relative Importance (IRI), which indicates the 117 importance of each prey category consumed. We calculated it by summing the percentages of 118 number (N%), frequencyNorth-Western (F%), and volume (V%) (as proposed in Biavati et al. 2004): accepted 119 120 To calculate the trophic niche breadth, we used the Levins index (B) described by 121 Pianka (1986), where p is the numerical or volumetric proportion of prey category i and n is 122 the number of prey categories: 123 Use the following type of citation: North-western Journal of Zoology 2021: e211508 nwjz-6 124 The index (B) varies from 1 (specialists in use of food resource) to n (generalists in the 125 use of resources). For better interpretation, we standardized Levin’s measure of niche breadth 126 (Bsta), which limits the value on a scale from 0 to 1 according to the equation below, where n 127 represents the number of resources (prey species) registered. Niche breadth values from 0.0 to 128 0.50 we interpreted as a specialist diet, and values from 0.51 to 1.0 – as a generalist (Hurlbert 129 1978). 130 131 We removed the oocytes from the abdominal cavity of gravid females and preserved 132 them in 5% formalin. We counted the number of oocytes of each female to represent the 133 clutch size. The diameter of 30 oocytes per female were measuredZoology using a stereomicroscope 134 with a millimetric ocular lens.
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