{"paper_id":"3705b306-53a5-4e72-8207-19c2ec10b959","body_text":"PREPRINT\nAuthor-formatted, not peer-reviewed document posted on 25/02/2022\nDOI: https://doi.org/10.3897/arphapreprints.e82751\nPresence of an alien turtle accelerates hatching of \ncommon frog (Rana temporaria) tadpoles\nMagda Vodrážková, Irena Šetlíková, Josef Navrátil, Michal Berec\n\n1 \n \nPresence of an alien turtle accelerates hatching of common frog (Rana temporaria ) 1 \ntadpoles 2 \nM. Vodrážková • I. Šetlíková • J. Navrátil • M. Berec 3 \nFaculty of Agriculture, University of South Bohemia, Studentská 1668, 370 05 České 4 \nBudějovice, Czech Republic 5 \n 6 \n*Corresponding author e-mail: Vodram02@zf.jcu.cz 7 \n 8 \nORCID of the authors: 9 \nMV: 0000-0002-2600-0346, IŠ: 0000 -0001-9993-4978, JN: 0000 -0002-4600-1012 and MB: 10 \n0000-0002-1065-7343 11 \n 12 \nAbstract 13 \nThe presence of a predator affects prey populations either by direct pr edation or by modifying 14 \nvarious parts of their life history. We investigated whether the hatching time, developmental 15 \nstage, and body size at hatching of common frog (Rana temporaria) embryos would alter in the 16 \npresence of a red -eared slider ( Trachemys scripta elegans) as a predator. The presence of a 17 \npredator affected all factors examined. We found that in the absence of the slider, the embryos 18 \nhatched in 12 days, while hatching was accelerated by two days in slider treatment. At the same 19 \ntime, the embr yos hatched smaller and at a lower stage of development with the slider than 20 \nwithout it. Our study extends the range of predators studied, including the effect on different 21 \nphases of development of potential amphibian prey. 22 \n 23 \nAuthor-formatted, not peer-reviewed document posted on 25/02/2022. DOI:  https://doi.org/10.3897/arphapreprints.e82751\n\n2 \n \nIntroduction 24 \nThe impacts of invasive species on native communities are still difficult to generalise due to the 25 \nlimited number of species and environments researched (Griesemer et al. 2018; Ramírez 26 \nAlbores et al. 2019; Rolim et al. 2015; Tricarico et al. 2016). However, inappropriate responses 27 \nof individuals to invasive predators can strongly affect native populations (Mooney and Cleland 28 \n2001). In amphibians, predation can account for a significant proportion of the total mortality 29 \nof all their developmental stages (Gunzburger and Travis 2005; Chivers et al. 2001; Laurila et 30 \nal. 2002; Nyström et al. 1997) . The ability to detect, recognise, and respond to potential 31 \npredators is, therefore, an important part of antipredatory behaviour (Bennett et al. 2013; Polo‐32 \nCavia and Gomez‐Mestre 2014), and native populations can have especially serious problems 33 \nfacing the presence of new alien predators (Gomez-Mestre and Díaz-Paniagua 2011; Nunes et 34 \nal. 2019; Polo -Cavia et al. 2010) . In general, embryonic and early larval stages are the most 35 \nvulnerable to predation (Laurila et al. 2002; Wells 2007) , and the ability to respond to the 36 \npresence of a predator can therefore significantly increase the fitness of an individual and thus 37 \nthe viability of the entire population (Vonesh and Bolker 2005; Warkentin 1995). 38 \nWhether intentionally or unintentionally introduced, the recent wide occurrence of the 39 \nred-eared slider ( Trachemys scripta elegans ) in Europe (GISD 2021) presents a new 40 \nopportunity to investigate the responses of naive native amphibian populations to a new 41 \npredator. Although red -eared slider (hereafter referred to as slider) is not reproductively 42 \nsuccessful throughout Europe (Cadi et al. 2004; Ficetola et al. 2009; Mikátová and Šandera 43 \n2015; Standfuss et al. 2016), even the mere presence of adults may pose a certain risk to native 44 \nspecies. In previous studies, we found that the presence of the sliders affect several life history 45 \nparameters of common frog (Rana temporaria) tadpoles, such as movement activity, trajectory 46 \nof movement (Berec et al. 2016), time to metamorphosis, or size at metamorphosis (Vodrážková 47 \net al. 2020). Although sliders are usually still hibernating at the time of common frog breeding 48 \nAuthor-formatted, not peer-reviewed document posted on 25/02/2022. DOI:  https://doi.org/10.3897/arphapreprints.e82751\n\n3 \n \n(Gibbons et al. 1990; Speybroeck et al. 2016) , which eliminates the risk of direct predation, 49 \nkairomones released by sliders into the aquatic environment provide amphibians with 50 \ninformation about their presence. Since the slider is an opportunistic predator and can consume 51 \nfrog eggs (Ernst and Lovich 2009), some response of common frog embryos is to be expected. 52 \nFor frog embryos, there are two basic strategies for avoiding predation or significantly 53 \nreducing its effects: the development of egg unpalatability and hatching plasticity (Wells 2007). 54 \nThe unpalatability of eggs is a passive strategy in which the embryo relies on the predator's 55 \ninability or unwillingness to consume eggs, which imposes costs on its host even if the host 56 \nnever comes in contact with the predator; environmentally cued hatching is characterised by an 57 \nembryo’s active capability to alter the time of hatching according to the conditions it encounters 58 \nduring embryonic development. Hatchi ng plasticity has been documented many times in 59 \namphibian embryos, and predator presence has been shown to trigger early hatching from eggs 60 \nincubated in both air and water (Chivers et al. 2001; Warkentin 2011). In terrestrially laid eggs, 61 \nhatching can be stimulated by vibrational cues during the direct physical attacks of predators, 62 \nsuch as snakes (Jung et al. 2019; Warkentin 1995) , frogs (Vonesh and Bolker 2005), katydids 63 \n(Poo and Bickford 2014), wasps (Warkentin 2000), or egg-eating fly larvae (Vonesh and Bolker 64 \n2005). In aquatic environments, these responses are induced mainly by chemical cues from 65 \npredators (kairomones) or by chemical cues that are released from injured prey during predation 66 \nevents (Dodson 1988; Laurila et al. 2002; Nicieza 1999; 2000; Petranka et al. 1987; Smith and 67 \nFortune 2009; Tollrian 1994). 68 \nThis study aimed to shift our previous focus (Berec et al. 2016; Vodrážková et al. 2020; 69 \nin review)  to a different developmental stage, namely, embryos in eggs. We investigated 70 \nwhether the presen ce of a slider can alter the hatching time of common frog embryos. We 71 \nhypothesised that the presence of a slider would accelerate the hatching time, so the ontogenetic 72 \nstage and body size at hatching were also measured. The uniqueness of this study lies in the use 73 \nAuthor-formatted, not peer-reviewed document posted on 25/02/2022. DOI:  https://doi.org/10.3897/arphapreprints.e82751\n\n4 \n \nof a stage-nonspecific predator, which is virtually absent in the literature. At the same time, it 74 \nis an alien predator from a taxonomic group to which the prey has no common history. 75 \n 76 \nMaterials and methods 77 \nFive freshly laid clutches of common fro gs were collected in a pool  between Holubov and 78 \nVrábče, South Bohemia, the Czech Republic  (48.9078633N, 14.3485608E), on 2 April 2021. 79 \nCollection locality was  monitored daily to collect egg clutches laid during the night before. 80 \nNeither the slider nor any other species of turtle occurs at the collection locality, so the eggs 81 \nand their parents are naive prey relative to the turtles.  The experiment was performed in six 82 \nglass tanks – three replications with the sliders and three repl ications of control . Glass tanks 83 \n(size: 1 00 × 55 × 50  cm) filled with 20  cm of  aged tap water  were equipped with a Claro 84 \n300 filter pump (300 L.h −1) and rinsed three times a week. The room temperature was set at 85 \n15 °C and the datalogger (Dostman LOG200 PDF) recorded a mean air temperature of 14.8 ± 86 \n0.4 °C (± S.D.; measured at hourly intervals)  during the experiment . Fluorescent tubes (2 x 87 \n36 W) with a light regime of 12  h/12 h were used. During the dark phase of the day, the glass 88 \ntanks were illuminated with red light to allow permanent monitoring of egg hatching. 89 \nThree adult sliders (carapace length: 18 cm, 20 cm, and 21 cm) were used as predators. 90 \nThe slider was placed in each of three glass tanks three days to release kairomones into the 91 \nwater before the experiment was initiated and fed three times a week with ReptoMin Tetra turtle 92 \ngammarus. To prevent physical but not chemical contact between the slider and frog eggs, a 93 \nglass barrier was placed inside each glass tank with a 6 cm gap at both ends so that water could 94 \nflow freely throughout the tank. On the other side of this barrier, five perforated opaque boxes 95 \n(20 × 14 cm) with holes 1 mm in diameter were glued to the bottom of the glass tanks to contain 96 \nthe eggs (Fig. 1). 97 \nAuthor-formatted, not peer-reviewed document posted on 25/02/2022. DOI:  https://doi.org/10.3897/arphapreprints.e82751\n\n5 \n \n 98 \nFigure 1. 99 \nDiagram of the glass tank showing the position of the slider (if present) and the boxes for clutch 100 \nsamples. These were placed randomly in the boxes in each glass tank (see Materials and 101 \nmethods). Three replications with the sliders and three replications without them (control) were 102 \nused. Slider drawing by Jakub Berec. 103 \n 104 \nSix samples of approximately 150 eggs each were taken from the collected clutch and 105 \nrandomly placed in six boxes, one in each glass tank. This procedure was repeated for all five 106 \nclutches, so that there were five boxes in each tank with sample from each clutch.  Each glass 107 \ntank was continuously monitored using a camera (Niceboy Stream Pro). Hatched tadpoles were 108 \ncounted every 24 h. Hatching was defined as the moment at which the whole hatchling had left 109 \nthe protective jelly of the eggs . To maintain a good processing of the camera recording s (the 110 \nlarge number of hatched tadpoles  in a small box makes it difficult to count them ), hatched 111 \ntadpoles were transferred every six hours to a depot tank. At the time when half of the eggs in 112 \neach box had hatched, two tadpoles were taken from the group of tadpoles hatched in the last 113 \nAuthor-formatted, not peer-reviewed document posted on 25/02/2022. DOI:  https://doi.org/10.3897/arphapreprints.e82751\n\n6 \n \nsix hours. These tadpoles were photographed under a stereomicroscope (Olympus SZX 7) and 114 \nmeasured (to the nearest 0.01 mm) using QuickPHOTO MICRO  3.2 software. Their 115 \ndevelopmental phase was determined according to Gosner (1960). 116 \nThe experiment involved a four factor design (slider: presence/absence, glass tank: 1-3 117 \nwith slider, 4-6 controls, box: five in each glass tank, and clutch: six samples). The slider was 118 \nused as a fixed factor as both levels of this factor (presence/absence) were tested. All other three 119 \nfactors (glass tank, clutch, and box) were random (Allen 2017) with the glass tank factor nested 120 \nin the slider presence/absence factor. According the experimental design, linear mixed model 121 \nwas used for analysis (Quinn and Keough 2002). Three analyses were performed – for hatching 122 \ntime, developmental stage, and the size at hatching. Adjusted R -squared was used as measure 123 \nof variability explained in the statistical model. Effect sizes were evaluated by  partial eta -124 \nsquared (Richardson 2011). Given the number of eggs, statistical significance was assessed at 125 \nthe 99.9% level (Steel et al. 2013). All calculations were done in Tibco Statistica (TIBCO 126 \n2017). 127 \n 128 \nResults 129 \nAll three models for life history parameters measured were statistically significant (hatching 130 \ntime: F=688.7, p<<0.001; adjusted R2=0.749; developmental stage: F=27.1, p<<0.001; adjusted 131 \nR2=0.852; size at hatching: F=23.6 , p<<0.001; adjusted R 2=0.833). For all these parameters, 132 \nthe presence of the slider was the only significant or far most important factor in each mo del 133 \n(Supplementary file: partial eta-squared in Tables 1-3). 134 \nWe found a significant difference in hatching time between the presence and absence of 135 \nthe slider (F(1,4)=915.0; p<< 0.001). In the absence of the slider, embryos hatched in 136 \n12 ± 0.6 days (mean ± S.D.). The presence of the sli der accelerated hatching by two  days 137 \n(10 ± 0.6 days) (Fig. 2). Hatching time differed significantly also among glass tanks 138 \nAuthor-formatted, not peer-reviewed document posted on 25/02/2022. DOI:  https://doi.org/10.3897/arphapreprints.e82751\n\n7 \n \n(F(1,4)=9.5; p<<0.001), boxes (F(1,4)=7.6; p<< 0.001) and clutches (F(1,4)=44.3; p<< 0.001), 139 \nbut the effect sizes of these three factors were negligible in comparison to the effect of slider 140 \npresence (Supplementary file: partial eta-squared in Table 1).  141 \nSimilarly, significant differences were found between the developmental stage 142 \n(F(1,4)=4608.0; p<0.01) and size (F(1,4)=75.1; p< 0.001) of freshly hatched embryos in the 143 \npresence of the slider and without it. In the presence of a slider, embryos hatched at 144 \ndevelopmental stage 20 ± 1.5 (mean ± S.D.) with an average size of 5.92 ± 1.460 mm, while in 145 \nthe control, freshly hatched embryos had developed to stage 23  ± 1.0, with an average size of 146 \n10.77 ± 1.042 mm (Fig. 2). Neither developmental stage nor hatching size was significantly 147 \naffected by glass tank, clutch and boxes (Supplementary file: Tables 2 and 3). 148 \n 149 \nFigure 2. 150 \nHistogram of A hatching time, B Gosner (developmental) stage, and C size at hatching of the 151 \nembryos of common frogs in the presence of red-eared slider and control. 152 \n 153 \nDiscussion 154 \nHatching Time (days)\n8 9 10 11 12 13\n0\n20\n40\n60\n80\nNumber of Individuals (%)\n0\n20\n40\n60\n80\n100\nDevelopmental Stage\n18 19 20 21 22 23 24\nSize at Hatching (mm)   < 4\n   4-5\n   5-6\n   6-7\n   7-8\n   8-9\n   9-10\n   10-11\n   11-12\n   > 12\nControl\nRed-eared Slider\nA B C\nAuthor-formatted, not peer-reviewed document posted on 25/02/2022. DOI:  https://doi.org/10.3897/arphapreprints.e82751\n\n8 \n \nDevelopmental plasticity is an adaptive response of anuran embryos and larvae to the risk of 155 \npredation (Altig and McDiarmid 1999; Benard 2004; Warkentin 2011) . Here, we present 156 \nevidence for the developmental plasticity of common frog embryos in the presenc e of a red -157 \neared slider and, in addition to a previous study (Vodrážková et al. 2020), provide a 158 \ncomprehensive insight of the influence of this alien predator on the early phases of the common 159 \nfrog life cycle. We have previously shown (Vodrážková et al. 2020) that, in the slider presence, 160 \ntadpoles of common frogs are able to modify the duration of larval development. In the present 161 \nstudy, we confirmed a similar response in common frog embryos, which hatched earlier in the 162 \npresence of a slider. At the same time, the embryos were smaller and less developed when 163 \nexposed to the chemical signals of a predator. We also found the effect of glass tank, clutch and 164 \nbox on hatching time, which was nevertheless negligible in comparison with t he effect of 165 \npredator presence. 166 \nIn the presence of stage -specific predators, amphibians can modify the duration of the 167 \nrelevant developmental stage (Chivers et al. 2001; Ireland et al. 2007; Mitchell et al. 2017). In 168 \nanuran embryos, specifically, the presence of egg predators has mostly been shown to induce 169 \nearly hatching of embryos (Chivers et al. 2001; Johnson et al. 2003; Laurila et al. 2001; Segev 170 \net al. 2015; Warkentin 1995; 2000) , while tadpole predators induce delayed hatching (Laurila 171 \net al. 2002; Mitchell et al. 2017; Schalk et al. 2002; Sih and Moore 1993), thus increasing their 172 \nchance of survival by escaping possibl e attacks. However, the slider is not a stage -specific 173 \npredator, as it is capable of consuming both amphibian eggs and larvae (Brown et al. 1995; 174 \nErnst and Lovich 2009; Chen 2006); thus, the allocation of risk between developmental stages 175 \nof the frog may b e more complex in this case (Warkentin 2011). Studies examining predator 176 \neffects on the developmental rates of both eggs and larvae are rare because few predators 177 \nconsume both eggs and larvae simultaneously. Muraro et al. (2021)  used a stage -nonspecific 178 \npredator ( Procambarus clarkii ) and found, in concordance with our results, a reduction in 179 \nAuthor-formatted, not peer-reviewed document posted on 25/02/2022. DOI:  https://doi.org/10.3897/arphapreprints.e82751\n\n9 \n \nhatching time in Rana latastei  embryos. However, they did not study larval development. 180 \nIreland et al. (2007) solved the problem of predator stage specificity by simult aneously 181 \nexposing frog eggs to stage-specific predators of eggs (leech: Nephelopsis obscura) and larvae 182 \n(dragonfly: Aeshna canadensis nymphs), which resulted in no change in hatching time, whereas 183 \ntests with separately acting predators produced the expected response of a reduction in hatching 184 \ntime in the egg predator treatment and an increase in hatching time in the  larval predator 185 \ntreatment. This study on embryos and a previous study on tadpoles (Vodrážková et al. 2020)  186 \njointly clarify that the embryos/tadpoles of the common frog  responded to the presence of a 187 \npredator by shortening the stage of development during which the embryo/tadpole would be 188 \nexposed to the predator. It would be interesting to analyse how common frog tadpoles react to 189 \nthe presence of a slider if the entire development from eggs to metamorphosis was taking place 190 \nwith this predator present. 191 \nHowever, some studies have shown that frog embryos, including the common frog, do 192 \nnot always respond specifically to stage -specific predators by short ening hatching time 193 \n(Capellán and Nicieza 2010; Laurila et al. 2001; Laurila et al. 2002; Saglio and Mandrillon 194 \n2006; Schalk et al. 2002; Touchon et al. 2006; Touchon and Wojdak 2014) . The published 195 \ndifferences in embryo responses may correspond to differe nt signal intensities of the presence 196 \nof a specific predator, and thus, the responses to indirect waterborne cues might be weaker than 197 \nthose to the direct, mechanical cues of a predator attack (Warkentin 2011). An evident response 198 \nto water-borne cues of sl iders may be related to a markedly stronger signal of a much larger -199 \nsized predator in our experiment compared to commonly tested invertebrate predators. The 200 \nability to scale predator danger and adjust hatching time accordingly has been found, for 201 \nexample, in embryos of southern leopard frogs ( Lithobates sphenocephalus) (Johnson et al. 202 \n2003). Moreover, a possible absence of a change in hatching time does not necessarily imply a 203 \ncomplete lack of response to the presence of a predator. It may be manifested by other types of 204 \nAuthor-formatted, not peer-reviewed document posted on 25/02/2022. DOI:  https://doi.org/10.3897/arphapreprints.e82751\n\n10 \n \nresponses, such as changes in the body shape of tadpoles (Laurila et al. 2001; Mandrillon and 205 \nSaglio 2007; Saglio and Mandrillon 2006; Touchon and Wojdak 2014)  or their behaviour 206 \n(Saglio and Mandrillon 2006; Touchon and Wojdak 2014). 207 \nNative and naive prey may fail to detect the novel predator adequately as a dangerous 208 \nthreat, resulting in no (Cox and Lima 2006; Sih et al. 2010) or inefficient antipredator responses 209 \nto counter the predator’s attack strategies (Sih et al. 2010; Strauss et al. 2006). However, when 210 \nresponses in hatching time in naive prey are detected, they are often explained by the presence 211 \nof syntopic, taxonomically related predators (Melotto et al. 2021; Muraro et al. 2021; Sih et al. 212 \n2010), although the time since invasion ma y also play an important role (Gomez-Mestre and 213 \nDíaz-Paniagua 2011; Nunes et al. 2013) . Our results suggested that a common evolutionary 214 \nhistory is not necessary for a detectable response. Such a result has already been published for 215 \ntadpole development time (Stav et al. 2007; Vodrážková et al. 2020), but as far as we know, it 216 \nhas not yet been published for hatching time in frog embryos.  An explanation for embryo 217 \nresponse to an alien slider may be in the ability of embryos to detect a kind of general \"smell  218 \nof fear\" that is elicited by most predators, regardless of taxonomic classification (Sih et al. 219 \n2010). 220 \nFinding a general tendency in the phenotypic plasticity responses of prey across a broad 221 \nrange of animal predators (different taxa and feeding spectra), environmental and experimental 222 \nconditions is a challenge even in anurans themselves ( Relyea et al. 2018 ). However, in frogs, 223 \nthe earlier hatching time was generally associated with smaller size at hatching (Capellán and 224 \nNicieza 2007 ; Chivers et al. 2001 ; Ireland et al. 2007 ; Laurila et al. 2002) and lower 225 \ndevelopmental stage (Capellán and Nicieza 2007 ; Chivers et al. 2001 ; Ireland et al. 2007 ; 226 \nLaurila et al. 2002 ; Muraro 2021), and our results confirm this relationship. In some cases, 227 \nearlier hatched tadpo les performed higher growth rate and reached the size of later ha tched 228 \ntadpoles at metamorphosis (Capellán and Nicieza 2007 ). However, if tadpoles are unable to 229 \nAuthor-formatted, not peer-reviewed document posted on 25/02/2022. DOI:  https://doi.org/10.3897/arphapreprints.e82751\n\n11 \n \ncompensate for their smaller size at hatching, this can impose significant costs in later 230 \ndevelopmental phases. These costs have been demonstrated in increased mortality during the 231 \nlarval stage (Smith 1987; Warkentin 1995; but see Vonesh and Bolker (2005) where early 232 \nhatchlings survived better), reduced size at metamorphosis (Vonesh and Bolker 2005;  233 \nVodrážková et al. 2020), lower post -metamorphic survival (Altwegg and Reyer 2003; Berven 234 \n1990), change of behaviour (Buckley et al. 2005, Capellán and Nicieza 2007), delayed maturity 235 \n(Smith 1987) and lower reproductive success (Smith 1987).  236 \nOur work added a slider as an additional  predator inducing changes in the embryonic 237 \ndevelopmental rate in Ranidae. Since the impact of earlier embryo hatching (lower body size 238 \nand lower stage of development) on fitness has been confirmed in several frog species (Laurila 239 \net al. 2002; Touchon et al. 2013; Vonesh and Bolker 2005; Warkentin 1995) , the same impact 240 \ncan be expected for the common frog. T he existence of defensive responses in slider -exposed 241 \nembryos may reduce the threat that poses the spreading of this inva sive species in Europe. On 242 \nthe other hand, the reduced size at hatching and developmental stage of common frog hatchlings 243 \nrepresents additional risks of negative fitness impacts, and at the very least, the presence of 244 \nsliders in non-native areas should receive increased attention. 245 \n 246 \nFunding/Acknowledgements/Competing interests 247 \nM.V. acknowledges GAJU 045/2019/Z for financial support. The authors have no support to 248 \nreport. The authors have declared that no competing interests exist. 249 \nAll methods were carried ou t in accordance with relevant guidelines and regulations. All 250 \nexperimental protocols were approved by the Czech Ministry of Agriculture, Department of 251 \nAnimal Welfare according to article No.  15, section  2 of the act registered under number 252 \n9103/2009-17210. 253 \n 254 \nAuthor-formatted, not peer-reviewed document posted on 25/02/2022. DOI:  https://doi.org/10.3897/arphapreprints.e82751\n\n12 \n \nReferences 255 \nAllen M. (2017) The sage encyclopedia of communication research methods (Vols. 1-4). SAGE 256 \nPublications, Inc., (Thousand Oaks, CA): 1-2064. 257 \nAltig R,  McDiarmid RW (1999) Tadpoles: The biology of anuran larvae. University of Chicago 258 \nPress, (Chicago): 1-454. 259 \nAltwegg, R., & Reyer, H. U. (2003). Patterns of natural selection on size at metamorphosis in 260 \nwater frogs. Evolution, 57, 872 -882. https://doi.org/10.1111/j.0014-261 \n3820.2003.tb00298.x 262 \nBenard MF (2004) Predator -induced phenotypic plasticity in organisms with complex life 263 \nhistories. Annual Review of Ecology, Evolution, and Systematics 35: 651 -673. 264 \nhttps://doi.org/10.1146/annurev.ecolsys.35.021004.112426. 265 \nBennett AM, Pereira D,  Murray DL (2013) Investment into defensive traits by anuran prey 266 \n(Lithobates pipiens) is mediated by the starvation-predation risk trade-off. PLoS One 8: 267 \ne82344. https://doi.org/10.1371/journal.pone.0082344. 268 \nBerec M, Klapka V,  Zemek R (2016) Effect of an alien turtle predator on movement activity 269 \nof European brown frog tadpoles. Italian Journal of Zoology 83: 68 -76. 270 \nhttps://doi.org/10.1080/11250003.2016.1139195. 271 \nBerven, K. A. (1990). Factors affecting population fluctuations in larval and adult stages of the 272 \nwood frog (Rana sylvatica). Ecology, 71, 1599-1608. https://doi.org/10.2307/1938295 273 \nBrown HA, Bury RB, Darda DM, Diller L, Peterson C, Storm R (1995) Reptiles of Washington 274 \nand Oregon. Seattle Audubon Society, (Seattle, WA): 1-176. 275 \nBuckley, C. R., Michael, S. F., & Irschick, D. J. (2005). Early hatching decreases jump ing 276 \nperformance in a direct-developing frog, Eleutherodactylus coqui. Functional ecology, 277 \n67-72.  278 \nCadi A, V D, Prévot‐Julliard AC, Joly P, Pieau C, Girondot M (2004) Successful reproduction 279 \nof the introduced slider turtle ( Trachemys scripta elegans ) in the  South of France. 280 \nAquatic Conservation: Marine and Freshwater Ecosvstems 14: 237 -246. 281 \nhttps://doi.org/10.1002/aqc.607. 282 \nCapellán E,  Nicieza AG (2007) Trade-offs across life stages: does predator–induced hatching 283 \nplasticity reduce anuran post -metamorphic pe rformance? Evolutionary Ecology 21: 284 \n445-458. https://doi.org/10.1007/s10682-006-9133-9. 285 \nCapellán E,  Nicieza AG (2010) Constrained plasticity in switching across life stages: pre -and 286 \npost-switch predators elicit early hatching. Evolutionary Ecology 24: 49 -57. 287 \nhttps://doi.org/10.1007/s10682-008-9289-6. 288 \nChen T-H (2006) Distribution and status of the introduced red-eared slider (Trachemys scripta 289 \nelegans) in Taiwan. In: Koike F, Cloud M, Kawamichi M, De Poorter M, Iwatsuki K 290 \n(Eds) Assessment and control of bio logical invasion risks. Shoukadoh Book Sellers 291 \n(Kyoto, Japan and IUCN, Gland, Switzerland): 187-195.  292 \nChivers DP, Kiesecker JM, Marco A, Devito J, Anderson MT, Blaustein AR (2001) Predator‐293 \ninduced life history changes in amphibians: egg predation induces h atching. Oikos 92: 294 \n135-142. https://doi.org/10.1034/j.1600-0706.2001.920116.x. 295 \nCox JG,  Lima SL (2006) Naiveté and an aquatic –terrestrial dichotomy in the effects of 296 \nintroduced predators. Trends in Ecology & Evolution 21: 674 -680. 297 \nhttps://doi.org/10.1016/j.tree.2006.07.011. 298 \nDodson S (1988) The ecological role of chemical stimuli for the zooplankton: Predator‐299 \navoidance behavior in Daphnia. Limnology and Oceanography 33: 1431 -1439. 300 \nhttps://doi.org/10.4319/lo.1988.33.6part2.1431. 301 \nErnst CH,  Lovich JE (2009) Turtles of the United States and Canada. JHU Press, (Baltimore): 302 \n1-840. 303 \nAuthor-formatted, not peer-reviewed document posted on 25/02/2022. DOI:  https://doi.org/10.3897/arphapreprints.e82751\n\n13 \n \nFicetola GF, Thuiller W,  Padoa‐Schioppa E (2009) From introduction to the establishment of 304 \nalien species: bioclimatic differences between presence and reproduction localities in 305 \nthe sli der turtle. Diversity and Distributions 15: 108 -116. 306 \nhttps://doi.org/10.1111/j.1472-4642.2008.00516.x. 307 \nGibbons J, Greene J,  Congdon J (1990) Temporal and spatial movement patterns of sliders and 308 \nother turtles. In: Gibbons J (Ed) Life history and ecology of the slider turtle. Smithsonian 309 \nInstitution Press (Washington, D.C.): 201-215.  310 \nGlobal Invasive Species Database. http://www.issg.org/database [accessed 14/10/2021] 311 \nGomez-Mestre I,  Díaz-Paniagua C (2011) Invasive predatory crayfish do not trigger inducible 312 \ndefences in tadpoles. Proceedings of the Royal Society B: Biological Sciences 278: 313 \n3364-3370. https://doi.org/10.1098/rspb.2010.2762. 314 \nGosner KL (1960) A simplified table for staging anuran embryos and larvae with notes on 315 \nidentification. Herpetologica 16: 183-190.  316 \nGriesemer J, Jeschke J,  Heger T (2018) Mapping theoretical and evidential landscapes in 317 \necological science: Levin's virtue trade -off and the hierarchy -of-hypotheses approach. 318 \nIn: Jeschke J, Heger T (Eds) Invasion biology: Hypotheses and evide nce. CABI 319 \n(Boston, MA): 23-29.  320 \nGunzburger MS,  Travis J (2005) Critical literature review of the evidence for unpalatability of 321 \namphibian eggs and larvae. Journal of Herpetology 39: 547 -571. 322 \nhttps://doi.org/10.1670/1-05A.1. 323 \nIreland D, Wirsing A,  Murray D (2007) Phenotypically plastic responses of green frog embryos 324 \nto conflicting predation risk. Oecologia 152: 162 -168. https://doi.org/10.1007/s00442-325 \n006-0637-3. 326 \nJohnson JB, Saenz D, Adams CK, Conner RN (2003) The influence of predator threat on the 327 \ntiming of a life-history switch point: predator-induced hatching in the southern leopard 328 \nfrog ( Rana sphenocephala ). Canadian Journal of Zoology 81: 1608 -1613. 329 \nhttps://doi.org/10.1139/z03-148. 330 \nJung J, Kim SJ, Pérez Arias SM, McDaniel JG, Warkentin KM (2019) How do red -eyed 331 \ntreefrog embryos sense motion in predator attacks? Assessing the role of vestibular 332 \nmechanoreception. Journal of Experimental Biology 222: jeb206052. 333 \nhttps://doi.org/10.1242/jeb.206052. 334 \nLaurila A, Crochet P -A,  Merilä J (2001) Predation -induced effects on hatchling morphology 335 \nin the common frog ( Rana temporaria ). Canadian Journal of Zoology 79: 926 -930. 336 \nhttps://doi.org/10.1139/z01-045. 337 \nLaurila A, Pakkasmaa S, Crochet P-A, Merilä J (2002) Predator-induced plasticity in early life 338 \nhistory and morphology in two anuran amphibians. Oecologia 132: 524 -530. 339 \nhttps://doi.org/10.1007/s00442-002-0984-7. 340 \nMandrillon A-L,  Saglio P (2007) Herbicide exposure affects the chemical recognition of a non 341 \nnative predator in common toad tadpoles ( Bufo bufo ). Chemoecology 17: 31 -36. 342 \nhttps://doi.org/10.1007/s00049-006-0354-8. 343 \nMelotto A, Ficetola GF, Alari E, Romagnoli S,   Manenti R (2021) Visual recognition and 344 \ncoevolutionary history drive responses of amphibians to an invasive predator. 345 \nBehavioral Ecology 32: 1352-1362. https://doi.org/10.1093/beheco/arab101. 346 \nMikátová B,  Šandera M (2015) První rozmnožování volně žijící želvy nádherné ( Trachemys 347 \nscripta) na území České republiky. Herpeta 1: 5-6.  348 \nMitchell MD, Bairos -Novak KR,  Ferrari MC (2017) Mechanisms underlying the control of 349 \nresponses to predator odours in aquatic prey. Journal of Experimental Biology 220: 350 \n1937-1946. https://doi.org/10.1242/jeb.135137. 351 \nAuthor-formatted, not peer-reviewed document posted on 25/02/2022. DOI:  https://doi.org/10.3897/arphapreprints.e82751\n\n14 \n \nMooney HA,  Cleland EE (2001) The evolutionary impact of invasive species. Proceedings of 352 \nthe National Academy of Sciences 98: 5446 -5451. 353 \nhttps://doi.org/10.1073/pnas.091093398. 354 \nMoore RD, Newton B,  Sih A (1996) Delayed hatching as a response of streamside salamander 355 \neggs to chemical cues from predatory sunfish. Oikos: 331 -335. 356 \nhttps://doi.org/10.2307/3546073. 357 \nMuraro M, Romagnoli S, Barzaghi B, Falaschi M, Manenti R, Ficetola GF (2021) Invasive 358 \npredators induce plastic and adaptive responses during embryo deve lopment in a 359 \nthreatened frog. NeoBiota 70: 69. https://doi.org/10.3897/neobiota.70.65454. 360 \nNicieza AG (1999) Context‐dependent aggregation in Common Frog Rana temporaria  361 \ntadpoles: influence of developmental stage, predation risk and social environment. 362 \nFunctional Ecology 13: 852-858. https://doi.org/10.1046/j.1365-2435.1999.00375.x. 363 \nNicieza AG (2000) Interacting effects of predation risk and food availability on larval anuran 364 \nbehaviour and development. Oecologia 123: 497 -505. 365 \nhttps://doi.org/10.1007/s004420000343. 366 \nNunes AL, Fill JM, Davies SJ, Louw M, Rebelo AD, Thorp CJ, Vimercati G, Measey J (2019) 367 \nA global meta-analysis of the ecological impacts of alien species on native amphibians. 368 \nProceedings of the Royal Society B 286: 20182528. 369 \nhttps://doi.org/10.1098/rspb.2018.2528. 370 \nNunes AL, Richter -Boix A, Laurila A, Rebelo R (2013) Do anuran larvae respond 371 \nbehaviourally to chemical cues from an invasive crayfish predator? A community-wide 372 \nstudy. Oecologia 171: 115-127. https://doi.org/10.1007/s00442-012-2389-6. 373 \nNyström P, Axelsson E, Sidenmark J, Brönmark C (1997) Crayfish predation on amphibian 374 \neggs and larvae. Amphibia -Reptilia 18: 217 -228. 375 \nhttps://doi.org/10.1163/156853897X00107. 376 \nPetranka JW, Kats LB,  Sih A (1987) Predator -prey interactions among fish and larval  377 \namphibians: use of chemical cues to detect predatory fish. Animal Behaviour 35: 420 -378 \n425. https://doi.org/10.1016/S0003-3472(87)80266-X. 379 \nPolo-Cavia N, Gonzalo A, López P, Martín J (2010) Predator recognition of native but not 380 \ninvasive turtle predators by n aïve anuran tadpoles. Animal Behaviour 80: 461 -466. 381 \nhttps://doi.org/10.1016/j.anbehav.2010.06.004. 382 \nPolo‐Cavia N,  Gomez‐Mestre I (2014) Learned recognition of introduced predators determines 383 \nsurvival of tadpole prey. Functional Ecology 28: 432-439. https://doi.org/10.1111/1365-384 \n2435.12175. 385 \nPoo S,  Bickford DP (2014) Hatching plasticity in a Southea st Asian tree frog. Behavioral 386 \nEcology and Sociobiology 68: 1733-1740. https://doi.org/10.1007/s00265-014-1781-0. 387 \nQuinn G P, Keough MJ (2002) Experimental Design a nd Data Analysis for Biologists. 388 \nCambridge University Press (Cambridge): 1-537. 389 \nRamírez Albores JE, Badano EI, Flores Rivas JD, Flores Flores JL, Yáñez Espinosa L (2019) 390 \nScientific literature on invasive alien species in a megadiverse country: advances and  391 \nchallenges in Mexico. https://doi.org/10.3897/neobiota.48.36201. 392 \nRelyea, RA, Stephens, PR, Barrow, LN, Blaustein, AR, Bradley, PW, Buck, JC, Chang, A, 393 \nCollins, JP, Crother, B, Earl, J,  Gervasi, SS (2018) Phylogenetic patterns of trait and 394 \ntrait plasticity evolution: Insights from amphibian embryos. Evolution 72: 663-678. 395 \nhttps://doi.org/10.1111/evo.13428. 396 \nRichardson JTE (2011) Eta squared and partial eta squared as measures of effect size in 397 \neducational research. Educational Research Review 6: 135 -147. 398 \nhttps://doi.org/10.1016/j.edurev.2010.12.001. 399 \nRolim RG, de Ferreira PMA, Schneider AA, Overbeck GE (2015) How much do we know 400 \nabout distribution and ecology of naturalized and invasive alien plant species? A case 401 \nAuthor-formatted, not peer-reviewed document posted on 25/02/2022. DOI:  https://doi.org/10.3897/arphapreprints.e82751\n\n15 \n \nstudy from subtropical southern Brazil. Biological Invasions 17: 1497 -1518. 402 \nhttps://doi.org/10.1007/s10530-014-0811-1. 403 \nSaglio P,  Mandrillon A -L (2006) Embryonic experience to predation risk affects tadpoles of 404 \nthe common frog ( Rana temporaria ). Archiv fur Hydrobiologie: 505 -523. 405 \n10.1127/0003-9136/2006/0166-0505. 406 \nSegev O, Rodríguez A, Hauswaldt S, Hugemann K, Vences M (2015) Flatworms ( Schmidtea 407 \nnova) prey upon embryos of the common frog ( Rana temporaria) and induce minor 408 \ndevelopmental acceleration. Amphibia -Reptilia 36: 155 -163. 10.1163/15685381 -409 \n00002992. 410 \nSchalk G, Forbes MR,  Weatherhead PJ (2002) Development al plasticity and growth rates of 411 \ngreen frog (Rana clamitans) embryos and tadpoles in relation to a leech ( Macrobdella 412 \ndecora) predator. Copeia 2002: 445 -449. https://doi.org/10.1643/0045-413 \n8511(2002)002[0445:DPAGRO]2.0.CO;2. 414 \nSih A, Bolnick DI, Luttbeg B, Orrock JL, Peacor SD, Pintor LM, Preisser E, Rehage JS, Vonesh 415 \nJR (2010) Predator –prey naïveté, antipredator behavior, and the ecology of predator 416 \ninvasions. Oikos 119: 610-621. https://doi.org/10.1111/j.1600-0706.2009.18039.x. 417 \nSih A,  Moore RD (1993) Delayed hatching of salamander eggs in response to enhanced larval 418 \npredation risk. The American Naturalist 142: 947-960.  419 \nSmith, D. C. (1987). Adult recruitment in chorus frogs: effects of size and date  at 420 \nmetamorphosis. Ecology, 68, 344-350. https://doi.org/10.2307/1939265 421 \nSmith GR,  Fortune DT (2009) Hatching plasticity of wood frog ( Rana sylvatica ) eggs in 422 \nresponse to mosquitofish ( Gambusia affinis ) cues. Herpetological Conservation and 423 \nBiology 4: 43-47.  424 \nSpeybroeck J, Beukema W, Bok B, Van Der Voort J (2016) Field guide to the amphibians and 425 \nreptiles of Britain and Europe. Bloomsbury Publishing, (London, UK): 1-434. 426 \nStandfuss B, Lipovšek G, Fritz U, Vamberger M (2016) Threat or fiction: is the pond slider 427 \n(Trachemys scripta) really invasive in Central Europe? A case study from Slovenia. 428 \nConservation Genetics 17: 557-563. https://doi.org/10.1007/s10592-015-0805-2. 429 \nStav G, Kotler BP,  Blaustein L (2007) Direct and indirect effects of dragonfly (Anax imperator) 430 \nnymphs on green toad ( Bufo viridis ) tadpoles. Hydrobi ologia 579: 85 -93. 431 \nhttps://doi.org/10.1007/s10750-006-0388-5. 432 \nSteel, E. A., M. C. Kennedy, P. G. Cunningham, and J. S. Stanovick. 2013. Applied statistics 433 \nin ecology: common pitfalls an d simple solutions. Ecosphere 4 :115. 434 \nhttp://dx.doi.org/10.1890/ES13-00160.1. 435 \nStrauss SY, Lau JA,  Carroll SP (2006) Evolutionary responses of natives to introduced species: 436 \nwhat do introductions tell us about natural communities? Ecology Letters 9: 357 -374. 437 \nhttps://doi.org/10.1111/j.1461-0248.2005.00874.x. 438 \nTIBCO. 2017, Tibco Statistica™  Quick Reference . 439 \nhttps://docs.tibco.com/pub/stat/13.3.0/doc/pdf/TIB_stat_13.3_quick_ref.pdf [accessed 440 \n6/2/2022] 441 \nTollrian R (1994) Fish-kairomone induced morphological changes in Daphnia lumholtzi (Sars). 442 \nArchiv fur Hydrobiologie: 69 -75. https://doi.org/10.1127/archiv-443 \nhydrobiol/130/1994/69. 444 \nTouchon J, Gomez-Mestre I,  Warkentin K (2006) Hatching plasticity in two temperate anurans: 445 \nresponses to a pathogen and predation cues. Canadian Journal of Zoology 84: 556-563. 446 \nhttps://doi.org/10.1139/z06-058. 447 \nTouchon JC, McCoy MW, Vonesh JR, Warkentin KM (2013) Effects of plastic hatching timing 448 \ncarry over through metamorphosis in red‐eyed treefrogs. Ecology 94: 850 -860. 449 \nhttps://doi.org/10.1890/12-0194.1. 450 \nAuthor-formatted, not peer-reviewed document posted on 25/02/2022. DOI:  https://doi.org/10.3897/arphapreprints.e82751\n\n16 \n \nTouchon JC,  Wojdak JM (2014) Plastic hatching timing by red-eyed treefrog embryos interacts 451 \nwith larval predator identity and sublethal predation to affect prey morphology but not 452 \nperformance. PLoS One 9: e100623. https://doi.org/10.1371/journal.pone.0100623. 453 \nTricarico E, J unqueira AO,  Dudgeon D (2016) Alien species in aquatic environments: a 454 \nselective comparison of coastal and inland waters in tropical and temperate latitudes. 455 \nAquatic Conservation: Marine and Freshwater Ecosystems 26: 872 -891. 456 \nhttps://doi.org/10.1002/aqc.2711. 457 \nVodrážková M, Šetlíková I,  Berec M (2020) Chemical cues of an invasive turtle reduce 458 \ndevelopment time and size at metamorphosis in the common frog. Scientific Reports 459 \n10: 1-6. https://doi.org/10.1038/s41598-020-64899-0. 460 \nVodrážková M, Šetlíková I,  Berec M (in review) Different time patterns of the presence of red-461 \neared slider influence the ontogeny dynamics of common frog tadpoles. Scientific 462 \nReports: x-x.  463 \nVonesh JR,  Bolker BM (2005) Compensatory larval responses shift trade‐offs associated with 464 \npredator‐induced hatching plasticity. Ecology 86: 1580 -1591. 465 \nhttps://doi.org/10.1890/04-0535. 466 \nWarkentin KM (1995) Adaptive plasticity in hatching age: a response to predation risk trade -467 \noffs. Proceedings of the National Academy of Sciences 92: 3507 -3510. 468 \nhttps://doi.org/10.1073/pnas.92.8.3507. 469 \nWarkentin KM (2000) Wasp predation and wasp -induced hatching of red -eyed treefrog eggs. 470 \nAnimal Behaviour 60: 503-510. https://doi.org/10.1006/anbe.2000.1508. 471 \nWarkentin KM (2011) Plasticity of hatching in amphibians: evol ution, trade-offs, cues and 472 \nmechanisms. Integrative and Comparative Biology 51: 111 -127. 473 \nhttps://doi.org/10.1093/icb/icr046. 474 \nWells KD (2007) The Ecology and Behavior of Amphibians. The University of Chicago Press, 475 \n(Chicago): 1-114. 476 \nAuthor-formatted, not peer-reviewed document posted on 25/02/2022. DOI:  https://doi.org/10.3897/arphapreprints.e82751","source_license":"CC-BY-4.0","license_restricted":false}