ISSN 1827-9635 (print) © Firenze University Press ISSN 1827-9643 (online) www.fupress.com/ah Acta Herpetologica 6(1): 19-25, 2011 Body size and reproductive characteristics of paedomorphic and metamorphic individuals of the northern banded newt (Ommatotriton ophryticus) Eyup Başkale1, Ferah Sayım2, Uğur Kaya2 1 Pamukkale University, Faculty of Science & Arts, Department of Biology, 20017 Denizli, Turkey. Cor- responding author. Email: ebaskale@pau.edu.tr 2 Ege University, Science Faculty, Section of Biology, Department of Zoology, Bornova-İzmir, 35100 Turkey. Submitted on: 2010, 10th July; revised on: 2011, 9th March; accepted on: 2011, 21st March. Abstract. Paedomorphs and metamorphs of the northern banded newt (Ommatotri- ton ophryticus) were compared with respect to body size and some reproductive traits. Paedomorphic females are smaller than metamorphic individuals of the same sex. Both morphs have a similar reproduction pathway in terms of egg number and incu- bation period, while hatching success and egg diameter appear to be different between morphs. Survival rates of two morphs were significantly different and calculated as 66% for paedomorphs, and 81% for metamorphs. On the other hand, both forms shared a common allometric slope of the SVL vs. number of eggs and egg diameter, and larger females tend to produce more and larger eggs. Keywords. Facultative paedomorphosis, reproductive output, fecundity, body size. In many urodele species, including European newts (especially in Mesotriton sp., Lissotriton spp.) there are populations with alternative life-history pathways: metamor- phosis vs. paedomorphosis (e.g. Duellman and Trueb, 1994). Most larvae transform into immature individuals which remain more or less terrestrial before reaching sexual matu- rity; however, some larvae attain sexual maturity with larval morphology. Such a discrete life-history polymorphism has both genetic and environmental components, and is usu- ally viewed as an adaptation to contrasted environments in time and space (e.g. Denoël et al., 2005). Environmental conditions causing paedomorphosis are multifactorial and vary among species, but are often related to unsuitable terrestrial conditions, such that an aquatic lifestyle would be more beneficial. Paedomorphic forms of northern banded newt Ommatotriton ophryticus (Berthold, 1846) were described in 2008 by Kaya et al. (2008) near Karasu in the vicinity of Sakarya, Turkey. This species ranges from the western Caucasus in southern Russia and Georgia, mailto:ebaskale@pau.edu.tr 20 E. Başkale, F. Sayım and U. Kaya through northwestern Armenia and northern Turkey west to the Bosphorus Strait. It has been recorded from near sea level (Tabbaria Lake) up to around 2,750m asl (in Turkey). It is found in coniferous, mixed and deciduous forests up to sub-alpine meadows. Reproduc- tion of this species occurs in lakes, ponds, large puddles, drainage canals, roadside ditches in meadows, slow-flowing streams and stream pools in open areas near or within forests. O. ophryticus is listed as Near Threatened (NT) by IUCN (2009). For conserving any tar- get species, firstly, we have to understand its life-history. Although, breeding biology is an important part of life-histories, it has not been well documented for O. ophryticus until now. For this reason, we aimed to describe some aspects of the reproductive biology of paedomorphic and metamorphic O. ophryticus specimens by quantifying clutch size, egg size, hatching success and female size-reproduc- tive output relationships. Location of the study site and its characteristics were described in Kaya et al. (2008). To collect paedomorphic individuals, the field studies were conducted between late Feb- ruary and early March. These months were previously reported as the breeding season of O. ophryticus (Andren, 1997; Kutrup et al., 2005). Both forms were monitored for three years (2007-2009), and each year they were sampled at least three times. Individuals were captured by dip nets. Even though, a total of 146 individuals were captured in 2007, there were no paedomorphic individuals. In 2008, 191 individuals were captured of which seven paedomorphic specimens, while in 2009 of 280 captured individuals six were paedomor- phic individuals. Capture probability of paedomorphic individuals was estimated as 0 in 2007, 0.04 in 2008 and 0.02 in 2009 breeding seasons for this population. All captured pae- domorphic individuals and randomly selected metamorphic newts were transported alive to our amphibia laboratory in covered plastic boxes. To compare reproductive output, 21 pairs of metamorphic females and males (16 pairs in 2008 and five pairs in 2009) randomly selected, were transported to our laboratory. The temperature of the laboratory was main- tained at a constant temperature of 20±1 oC with a light/dark schedule of 14/10 hours. Each paedomorphic and metamorphic female was weighed with an UFO TECH pre- cision balance and their snout-vent lengths (SVL) were measured with digital calipers fol- lowing Başoğlu and Özeti (1973). Then we placed them with a metamorphic male in sepa- rate 20 liter glass aquariums and some small-leaved aquatic plants were provided for egg deposition. Newts were fed three times a week with tubifex worms and earthworms. We changed the water of each aquarium every other day. Paedomorphs and metamorphs in captivity displayed courtship behaviour and then breeding activity. The metamorphic and paedomorphic forms generally started to lay eggs on the 3rd day of captivity. Eggs were collected and counted four times a day, over the course of the entire oviposition period. As many species of newts and salamanders are especially fond of consuming their own eggs (Dasgupta, 1996; Kuzmin, 1991), we moved and tried to rear these eggs separately from the parents’ aquaria. Eggs which were transferred in a separate container were checked three or four times a day. Instead of the evaporating water during the incubation period, clean water was added to aquaria. Newly hatched larvae were counted and moved in a separate aquarium, and the experiment was lasted. The mean ovum diameter was calculated by measuring 20 randomly chosen eggs (in 1st cleavages) from each individual under a stereomicroscope with an ocular micrometer. Hatching success (Survival Rate) was defined as the number of hatched larvae divided by 21Reproductive characteristics of the northern banded newt the number of eggs and multiplied by 100. Confidence interval of survival rates was calcu- lated as CI = 2 SQRT [q(1–q)/n], where q = 1-SR, SR is the survival rate and n is number of eggs at the beginning of the experiment (Miaud, 1994). Body size measurements and all reproductive output parameters were normally dis- tributed (Kolmogorov–Smirnov D test, all P > 0.05), thus allowing comparisons using parametric tests. Firstly, to explore the level of variation among females within morphs, analyses of one-way ANOVA were performed using SPSS 13.0 Version for Windows. To analyze the relationships between female body size and egg size, we used standard- ized major axis (SMA) analysis (Warton et al., 2006). The mean of egg diameter for each female was regressed to the SVL in SMA analysis. SMA analysis provide a better esti- mate of the line summarizing the relationship between two variables (i.e., the main axis along which two variables are correlated) to that of ordinary linear regression, because the residual variance is minimized in both “x” and “y” dimensions. The analysis also deter- mined the differences between the slopes obtained for each species or for each maternal tree, so that a significant P indicated differences between the slopes of the groups studied. We used the free software statistics package SMART vers. 2.0 (Falster et al. 2006). There was a strong positive correlation between female SVL and body mass in paedomorphic (Pearson’s, n = 13; r2 = 0.820; P < 0.01), and metamorphic forms (Pearson’s, n = 21; r2 = 0.600; P < 0.01). Paedomorphic females were significantly smaller and lighter than meta- morphic females (Table 1 and Fig. 1). Body size can be varied between paedomorphs and metamorphs depending on the population source (Denoël et al., 2009). For example, in some populations of newts (smooth, palmate and alpine newt) paedomorphic forms can be smaller than metamorphs or vice versa as well as both forms have approximately same size (Denoël et al., 2009; Kalezić et al., 1996). The most notable results of our study that the mean number of eggs of the paedo- morphs lesser than metamorphs, although, there are not significant differences the num- ber of eggs between paedomorphs and metamorphs. However, egg diameters showed sta- tistically difference between paedomorphs and metamorphs (Table 1 and Fig. 1). Gener- ally, fecundity parameters and mating success are related to body size (Semlitsch, 1985), but the relation appears to be different in different cases. Paedomorphic females produced lesser eggs than metamorphic counterparts (Semlitsch, 1985; Kalezić et al., 1996), but, Table 1. Descriptive statistics of female size and reproductive outputs shown in two forms of O. ophryti- cus. Results of One way Anova indicate the differences between paeodomorphic and metamorphic forms. Form N Min Max Mean Std. Dev. df F Sig. SVL (mm) P 13 31.88 43.08 37.47 3.495 1 117.82 < 0.0001 M 21 44.96 64.34 52.68 4.232 Body mass (g) P 13 1.7 3.2 2.35 0.427 1 21.67 < 0.0001 M 21 2.4 6.4 3.70 0.985 Number of eggs P 13 16 189 83.46 55.611 1 3.59 0.067 M 21 25 296 122.90 60.965 Egg diameter (mm) P 116 0.60 0.98 0.78 0.068 1 314.41 < 0.0001 M 105 0.79 1.02 0.92 0.049 22 E. Başkale, F. Sayım and U. Kaya in some cases, the reverse situation also exists or they produced approximately the same number of eggs (Semlitsch, 1985; Kalezić et al., 1996; Rot-Nikcevic, et al., 2000). Totals of 1085 eggs of paedomorphic forms and 2581 eggs of metamorphic forms were layed, 717 and 2092 larvae of which developed, respectively. Photographs of an egg and a resulting larva are shown in Fig. 2. On the side, survival rates of 0.66 (66%), CI 0.63-0.69 for paedomorphs, and 0.81 (81%), CI 0.80-0.83 for metamorphs. These results indicate that survival rates of two morphs are significantly different each other. The growth and development rates of amphibians are influenced by a number of intrin- sic (body size or egg size and yolk reserves) and extrinsic factors (temperature, density and competition, food supply and quality, predation, breeding habitat and inhibitory compounds) (Crump, 1974, 1984; Kuramoto, 1975; Berven et al., 1979; Kaplan, 1980; Berven and Chadra, 1988; Duelman and Trueb 1994). Most likely, low survival rate in paedomorphic forms is underlined by intrinsic factors mostly because of constant labo- ratory conditions. The linear regression analysis indicated that female SVL showed a strong positive rela- tionship with egg diameter (metamorphs: r2 = 0.858; P = 0.024, paedomorphs: r2 = 0.823; P = 0.013), as well as with the number of eggs (metamorphs: r2 = 0.446; F = 15.232; P = 0.001, paedomorphs: r2 = 0.653; F = 20.713; P = 0.001). SMA analysis showed that both forms shared a common allometric slope of the SVL vs number of eggs and egg diameter. Pairwise combinations of SVL and number of eggs/egg diameters showed significant dif- ferences in elevation shift (Table 2). Consequently, our results indicate that larger individ- uals of both forms tend to produce more and larger eggs. Similarly, Furtula et al. (2008) found conformable relationships between the female SVL and vitellus volume in four crested newt species. In this sense, larger females produce larger eggs, and they have more yolk for larval development and/or thicker mucoid capsules which provide better protec- tion (Semlitsch, l985; Semlitsch and Gibbons, 1990; Kalezić et al., 1994). For this reason, female body size and/or egg size can affect the low survival rates in paedomorphic forms in contrast to metamorphic counterparts.   (a) S V L (m m ) 31 41 51 61 71 Paedomorphs Metamorphs (b)B od y M as s (g ) 1,7 2,7 3,7 4,7 5,7 6,7 Paedomorphs Metamorphs (c)N um be r of E gg s 0 50 100 150 200 250 300 Paedomorphs Metamorphs (d)E gg d ia m et er ( m m ) 0,59 0,69 0,79 0,89 0,99 1,09 1,19 Paedomorphs Metamorphs Fig. 1. Variations of SVL (a) and body mass (b), number of eggs (c) and egg diameter (d) in females of paedomorphic and metamorphic forms. 23Reproductive characteristics of the northern banded newt ACKNOWLEDGEMENTS We thank Dr. Milos Kalezić for his valuable comments on the manuscript, and also İ. Ethem Çevik and Şamil Yıldırım for assisting during field studies. The permissions for field work and han- dling of the newts were issued by the Animal Ethics Committee of Faculty of Medicine, Ege University.  Fig. 2. The eggs: fertilized (A), cleaved (B), and a resulting larva of paedomorphic form (C). Table 2. Results of standardized major axis (SMA) regression analysis for all pairwise combinations of snout-vent length (SVL) and reproduction traits of both forms. Trait pair (X and Y) Forms n r2 P Slope Intercept Slope homo- geneity (P) Shift in elevation (P) Shift along slope (P) SVL and egg number P 13 0.592 0.002 0.1286 1.333 0.877 0.000 < 0.0001 M 21 0.187 0.050 0.1340 1.454 SVL and Egg diameter P 6 0.342 0.223 18.47 4.288 0.409 0.004 0.009 M 5 0.062 0.687 34.57 2.589 24 E. Başkale, F. Sayım and U. Kaya REFERENCES Andren, C. (1997): On-the-spot appraisal at Hopa (Turkey). For threatened amphibians and reptiles 5-12 September, University of Göteborg, Sweden. Başoğlu, M., Özeti, N. (1973): Türkiye Amfibileri. Ege Üniv. Fen Fak. Kitaplar Serisi, İzmir 50: 155 p. Berven, K.A, Gill, D.E, Smith-Gill, S.J. (1979): Countergradient selection in the green frog, Rana clamitans. Evolution 33: 609–623. Berven, K.A., Chadra, B.G. (1988): The relationship among egg size, density, and food lev- el on larval development in the wood frog (Rana sylvatica). Oecologia 75: 67–72. Crump, M.L. (1974): Reproductive strategies in a tropical anuran community. Miscel. Publ. Univ. Kansas Mus. Nat. Hist. 61: 1–68. Dasgupta, R. (1996): Feeding ecology of the adult Himalayan Salamander Tylototriton ver- rucosus ANDERSON, 1871(Caudata: Salamandridae). Herpetozoa 9: 19–29. Denoël, M., Joly, P., Whiteman, H.H. (2005): Evolutionary ecology of facultative paedo- morphosis in newts and salamanders. Biol. Rev. 80: 663–671. Denoël, M., Ivanović, A., Džukić, G., Kalezić M.L. (2009): Sexual size dimorphism in the evolutionary context of facultativepaedomorphosis: insights from European newts. BMC Evol. Biol. 9: 278 Duellman, W., Trueb, L. (1994): Biology of amphibians. Baltimore and London, The Johns Hopkins University Press. Falster, D.S., Warton, D.I., Wright, I.J. (2006): SMATR: Standardised Major Axis Tests and Routines, vers. 2.0. Available at: http://www.bio.mq.edu.au/ecology/SMATR/. Furtula, M., Ivanović, A., Džukić, G., Kalezić, M.L. (2008): Egg size variation in Crested newts from the Western Balkans (Caudata: Salamandridae: Triturus cristatus Super- species). Zool. Stud. 47: 585–590. IUCN. (2009): IUCN Red list of threatened species. On line, URL: www.redlist.org. Kalezić, M.L., Cvetković, D., Djorovic, A., Džukić, G. (1994): Paedomorphosis and differ- ences in life-history traits of two neighboring crested newt (Triturus carnifex) popu- lations. Herpetol. J. 4: 151–158. Kalezić, M.L., Cvetkovic, D., Djorovic, A., Dzukic, G. (1996): Alternative life-history path- ways: paedomorphosis and adult fitness in European newts (Triturus vulgaris and T. alpestris). J. Zool. Syst .Evol. Res. 34: 1–7. Kaplan, R.H. (1980): The implications of ovum size variability for offspring fitness and clutch size within several populations of salamanders (Ambystoma). Evolution 34: 51–64. Kaya, U., Sayım, F., Başkale, E., Çevik, İ.E. (2008): Paedomorphosis in the banded newt, Triturus vittatus (Jenyns, 1835). Belg. J. Zool. 138: 196–197. Kuramoto, M. (1975): Embryonic temperature adaptation in development rate of frogs. Physiol. Zool. 48: 360– 366. Kutrup, B., Bulbul, U., Yilmaz, N. (2005): Age structure in two populations of Triturus vit- tatus ophryticus at different altitudes. Amphibia-Reptilia 26: 49–54. Kuzmin, S.L. (1991): The ecology and evolution of amphibian cannibalism. J. Bengal. Nat. Hist. Soc. (N.S.) 10: U-27. Miaud, C. (1994): Role of wrapping behaviour on egg survival in three species of Triturus (Amphibia: Urodela). Copeia 2: 535–537. 25Reproductive characteristics of the northern banded newt Rot-Nikcevic, I., Kalezić, M.L., Dzukic, G. (2000): Paedogenesis, life history traits and sex- ual dimorphism: a case study of the smooth newt, Triturus vulgaris, from Pannonia. Folia Zool. 49: 41–52. Semlitsch, R.D. (1985): Reproductive strategy of a facultatively paedomorphic salamander Ambystoma talpoideum. Oecologia 65: 305–313. Semlitsch, R.D., Gibbons, J.W. (1990): Effects of egg size on success of larval salamanders in complex aquatic environments. Ecology 71: 1789–1795. SPSS. (2004): SPSS 13.0 Base User’s Guide. SPSS Inc., Chicago, IL, USA. Warton, D.L., Wright, I.J., Falster, D.S., Westoby, M. (2006): Bivariate line-fitting methods for allometry. Biol. Rev. 81: 259–291. bbib67 OLE_LINK1 OLE_LINK2 bbib28 OLE_LINK5 OLE_LINK6 OLE_LINK7 OLE_LINK8 _GoBack OLE_LINK1 OLE_LINK2 OLE_LINK3 OLE_LINK4 OLE_LINK1 OLE_LINK2 OLE_LINK19 OLE_LINK20 OLE_LINK21 OLE_LINK29 OLE_LINK3 OLE_LINK4 OLE_LINK5 OLE_LINK31 OLE_LINK14 OLE_LINK15 OLE_LINK12 OLE_LINK13 OLE_LINK16 OLE_LINK17 OLE_LINK22 OLE_LINK23 OLE_LINK24 OLE_LINK8 OLE_LINK9 OLE_LINK10 OLE_LINK11 OLE_LINK18 OLE_LINK27 OLE_LINK28 OLE_LINK25 OLE_LINK26 OLE_LINK6 OLE_LINK7 OLE_LINK34 OLE_LINK37 OLE_LINK38 Acta Herpetologica Vol. 6, n. 1 - June 2011 Firenze University Press Widespread bacterial infection affecting Rana temporaria tadpoles in mountain areas Rocco Tiberti Extreme feeding behaviours in the Italian wall lizard, Podarcis siculus Massimo Capula1, Gaetano Aloise2 Lissotriton vulgaris paedomorphs in south-western Romania: a consequence of a human modified habitat? Severus D. Covaciu-Marcov*, Istvan Sas, Alfred Ş. Cicort-Lucaciu, Horia V. Bogdan Body size and reproductive characteristics of paedomorphic and metamorphic individuals of the northern banded newt (Ommatotriton ophryticus) Eyup Başkale1, Ferah Sayım2 , Uğur Kaya2 Genetic characterization of over hundred years old Caretta caretta specimens from Italian and Maltese museums Luisa Garofalo1, John J. Borg2, Rossella Carlini3, Luca Mizzan4, Nicola Novarini4, Giovanni Scillitani5, Andrea Novelletto1 The phylogenetic position of Lygodactylus angularis and the utility of using the 16S rDNA gene for delimiting species in Lygodactylus (Squamata, Gekkonidae) Riccardo Castiglia*, Flavia Annesi Localization of glucagon and insulin cells and its variation with respect to physiological events in Eutropis carinata Vidya. R. Chandavar1, Prakash. R. Naik2* The Balearic herpetofauna: a species update and a review on the evidence Samuel Pinya1, Miguel A. Carretero2 Effects of mosquitofish (Gambusia affinis) cues on wood frog (Lithobates sylvaticus) tadpole activity Katherine F. Buttermore, Paige N. Litkenhaus, Danielle C. Torpey, Geoffrey R. Smith*, Jessica E. Rettig Food composition of Uludağ frog, Rana macrocnemis Boulenger, 1885 in Uludağ (Bursa, Turkey) Kerim Çiçek Preliminary results on tail energetics in the Moorish gecko, Tarentola mauritanica Tommaso Cencetti1,2, Piera Poli3, Marcello Mele3, Marco A.L. Zuffi1 Climate change and peripheral populations: predictions for a relict Mediterranean viper José C. Brito1, Soumia Fahd 2, Fernando Martínez-Freiría1, Pedro Tarroso1, Said Larbes3, Juan M. Pleguezuelos4, Xavier Santos5 Assessing the status of amphibian breeding sites in Italy: a national survey Societas Herpetologica Italica* Osservatorio Erpetologico Italiano ACTA HERPETOLOGICA Journal of the Societas Herpetologica Italica ACTA HERPETOLOGICA Rivista della Societas Herpetologica Italica