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A description of the skeletal development of ranids is scarce despite being well known in the family Ranidae. Herein, the description of several species of two genera representing the family Ranidae from Turkey is studied wherein the larval and adult cranial skeletons of Pelophylax caralitanus are described and compared with that of the water frog (Pelophylax bedriagae) and mountain frog (Rana macrocnemis). The data are based on cleared and double-stained specimens of 20 Gosner developmental Stages (26-46). The first element to ossify in P. caralitanus is the parasphenoid (Stage 30), followed by the exoccipitals (Stage 34) and prootics (Stage 35). The metamorphic climax commences at Stage 42 during which several modifications to the chondrocranium will unfold. In addition, ranids demonstrate remarkable differences between the water (Pelophylax sp.) and mountain (Rana sp.) frog species: (a) the ventromedial bridge between the corpora of the suprarostral cartilage in the larval upper jaw, (b) the number of processes of the central corpus in the hyobranchial skeleton, and (c) the differences in the ossification time and sequence between Pelophylax sp. and Rana sp. A detailed description of the larval osteological features of P. caralitanus conforms to the current phylogenetic position and provides a model for comparison with other ranids., Tubitak (2218-National Postdoctoral Research Scholarship Programme), Tubitak (2218-National Postdoctoral Research Scholarship Programme).

Water striders (Hemiptera: Gerridae) are predators found on the water surface that prey mainly on arthropods. The feeding on other organisms (dead vertebrates, fishes and tadpoles) is a rare event. To our knowledge, predation of frog eggs by water striders has not yet been reported. We observed that adult water striders, Gerris latiabdominis Miyamoto, 1958 (Hemiptera: Gerridae), preyed on the eggs of three frog species, Pelophylax nigromaculata Hallowell, 1861 (Anura: Ranidae), Rana japonica Boulenger, 1879 (Anura: Ranidae), and Rhacophorus schlegelii Gunther, 1858 (Anura: Rhacophoridae). We found predation by the water striders affects the survival rate of frog eggs floating on the water surface. We suggest that this hunting event would occur in water bodies in which water striders and frogs coexist, especially the region where their phenology overlaps.

Mature spermatozoa traits and morphology of endangered Albanian water frog, Pelophylax shqipericus, have been characterized for the first time through phase contrast microscopy, as part of successful implementation of in vitro fertilization technique for this species. The basic morphology of P. shqipericus spermatozoa consists of an elongated, thick, smooth-edged, and solid-staining head, continuing with a thin and long tail which usually extends 2.48 times the head length. The acrosome was not clearly discernible so the measurements were done on the head as a whole, while the middle section was better visible. Average length of head, including the acrosome and midsection was estimated to be 11.78 μm ± 0.32, while the tail length resulted 29.24 ± 1.75 μm. The average thickness of the head was shown to be 3.45 μm. The total sperm length resulted to be 41.02 ± 1.83 μm. The average sperm concentration was estimated of 25.5 × 106 /ml. Sperm amount, survival rate and motility were also measured. The sperm survival rate was maximal immediately after preparation of the suspension and tended to decrease over time of storage, reaching 50% after 72 hr. Decreased sperm motility seemed to follow the same trend as sperm viability. Sperm traits resulted to be very similar both in size and in shape with those of "Lessonae" frog group, one of the lineages of Western Palearctic species complex, suggesting a strong phylogenetic relationship among these species.

Amphibians can be more vulnerable to environmental changes and diseases than other species because of their complex life cycle and physiological requirements. Therefore, understanding the adaptation of amphibians to environmental changes is crucial for their conservation. Major histocompatibility complex (MHC) presents an excellent tool for the investigation of adaptive variations and the assessment of adaptive potential because it can be under strong diversifying selection. Here, we isolated the MHC class II B (MHCIIB) gene from cDNA sequences of the black-spotted frog (Pelophylax nigromaculatus), a widespread amphibian species in China, and designed locus-specific primers to characterize adaptive variability of this amphibian. Ten alleles were identified from 67 individual frogs of three populations and no more than two alleles were present in each individual animal. Furthermore, none of the sequences had indels or/and stop codons, which is in good agreement with locus-specific amplification of a functional gene. However, we found low polymorphism at both nucleotide and amino acid levels, even in the antigen-binding region. Purifying selection acting at this locus was supported by the findings that the dN/dS ratio across all alleles was below 1 and that negatively selected sites were detected by different methods. Allele frequency distributions were significantly different among geographic populations, indicating that physiographic factors may have strong effect on the genetic structure of the black-spotted frog. This study revealed limited polymorphism of three adjacent black-spotted frog populations at the functional MHCIIB locus, which may be attributed to region-specific differences. The locus-specific genotyping technique developed in this study would provide a foundation for future studies on adaptive divergence among different frog populations.

Amphibians are often considered excellent environmental indicator species. Natural and man-made landscape features are known to form effective genetic barriers to amphibian populations; however, amphibians with different characteristics may have different species-landscape interaction patterns. We conducted a comparative landscape genetic analysis of two closely related syntopic frog species from central China, Pelophylax nigromaculatus (PN) and Fejervarya limnocharis (FL). These two species differ in several key life history traits; PN has a larger body size and larger clutch size, and reaches sexual maturity later than FL. Microsatellite DNA data were collected and analyzed using conventional (F-ST, isolation by distance (IBD), AMOVA) and recently developed (Bayesian assignment test, isolation by resistance) landscape genetic methods. As predicted, a higher level of population structure in FL (F-ST=0.401) than in PN (F-ST=0.354) was detected, in addition to FL displaying strong IBD patterns (r=.861) unlike PN (r=.073). A general north-south break in FL populations was detected, consistent with the IBD pattern, while PN exhibited clustering of northern- and southern-most populations, suggestive of altered dispersal patterns. Species-specific resistant landscape features were also identified, with roads and land cover the main cause of resistance to FL, and elevation the main influence on PN. These different species-landscape interactions can be explained mostly by their life history traits, revealing that closely related and ecologically similar species have different responses to the same landscape features. Comparative landscape genetic studies are important in detecting such differences and refining generalizations about amphibians in monitoring environmental changes.

The identification of the factors responsible for genetic variation and differentiation at adaptive loci can provide important insights into the evolutionary process and is crucial for the effective management of threatened species. We studied the impact of environmental viral richness and abundance on functional diversity and differentiation of the MHC class Ia locus in populations of the black-spotted pond frog (Pelophylax nigromaculatus), an IUCN-listed species, on 24 land-bridge islands of the Zhoushan Archipelago and three nearby mainland sites. We found a high proportion of private MHC alleles in mainland and insular populations, corresponding to 32 distinct functional supertypes, and strong positive selection on MHC antigen-binding sites in all populations. Viral pathogen diversity and abundance were reduced at island sites relative to the mainland, and islands housed distinctive viral communities. Standardized MHC diversity at island sites exceeded that found at neutral microsatellites, and the representation of key functional supertypes was positively correlated with the abundance of specific viruses in the environment (Frog virus 3 and Ambystoma tigrinum virus). These results indicate that pathogen-driven diversifying selection can play an important role in maintaining functionally important MHC variation following island isolation, highlighting the importance of considering functionally important genetic variation and host-pathogen associations in conservation planning and management.

Notwithstanding the abundance of amphibians, there are few descriptions about ranid cranial development. Herein, larval chondrocranial development of Uludag frog, Rana macrocnemis (Boulenger, 1885), is described on cleared and double-stained specimens. Descriptions are related with the ontogeny of the chondrocranium and osteogenesis of the cranial skeleton. The larval chondrocranial development of R. macrocnemis is compared to those of Rana and Pelophylax larvae (Pelophylax bedriagae, Rana pipiens, R. palustris, R. sphenocephala, R. catesbeiana, R. clamitans and R. sylvatica). In R. macrocnemis, the first bones to ossify are the parasphenoid and exoccipital (Stage 33), followed by the frontoparietal and prootic (stages 35 and 40, respectively). The major reconstruction of the chondrocranium begins at Stage 41. The ossification sequence of R. macrocnemis is distinguished from other ranids. Adult cranial osteology of R. macrocnemis is compared to that of P. bedriagae. Osteologically, R. macrocnemis is different from P. bedriagae by the shape and size of the vomer and number of teeth. Additionally, geometric morphometric methods are used to analyze chondrocranial size and shape changes of ranid larva of R. macrocnemis and P. bedriagae. Anat Rec, 299:711-721, 2016. © 2016 Wiley Periodicals, Inc.

Aim Pelophylax frogs in East Asia provide an opportunity to explore the impact of glacial cycling on demographic and genetic dynamics, because it has been suggested that they experienced distribution shifts and subsequent mtDNA introgression from Pelophylax plancyi to Pelophylax nigromaculatus in association with climatic oscillations. However, their evolutionary history, including the pattern of introgression, is incompletely understood. We used phylogenetic analyses based on multiple markers to address their evolutionary history, and palaeodistribution modelling to test whether the predicted distribution can explain the pattern of introgression suggested by molecular phylogenetics. Location East Asia, including far-eastern Russia, mainland China, the Japanese archipelago, the Korean Peninsula and Taiwan. Methods Ninety-nine samples of the Pelophylax nigromaculatus species complex were collected from 75 localities throughout the distributional range of each species. Phylogenetic analyses were performed with the mitochondrial cytochrome b (cytb) gene and six nuclear genes using two European Pelophylax frogs as outgroups. Additionally, palaeodistributions of P. nigromaculatus and P. plancyi were predicted using bioclimatic variables. Results Differences were observed between the mitochondrial and nuclear DNA trees in the phylogenetic position of P. plancyi. Similar phylogenetic discordance was also observed within P. nigromaculatus. Distribution modelling suggested that a considerable distributional shift of P. nigromaculatus occurred in association with glacial cycling and that the distributional range of P. plancyi was relatively narrow during glacial periods. Main conclusions The analyses detected five historical mtDNA introgression events and determined their directions, none of which had been deduced in previous studies. In association with glacial cycling, the distributional ranges of P. nigromaculatus and P. plancyi appear to have shifted repeatedly, resulting in multiple complex introgressions. By combining phylogenetic analyses with palaeodistribution modelling, our results supported the hypothesis that the history of mtDNA introgression among Pelophylax frogs was shaped by glacial cycling.

Previous studies have demonstrated that some amphibian species can be sex-reversed by high concentrations of androgens. Little attention has focused on the effects of androgenic endocrine-disrupting chemicals (EDCs) on amphibians. The present study aimed to investigate the effects of lower concentrations of the androgenic EDC 5α-dihydrotestosterone (DHT) on gonadal differentiation and development in Pelophylax nigromaculatus, a true frog distributed widely in East Asia. Tadpoles at Gosner stage 24/25 were exposed to nominal concentrations of 40 ng/L, 400 ng/L, and 4000 ng/L DHT to complete metamorphosis. In all DHT treatment groups, males and ambiguous sexes were identified based on gonadal morphology, whereas no females were found; thus, all treatment groups exhibited male-skewed ratios compared with the control group. Gonadal histological examination revealed that ambiguous sexes displayed overall testicular structure with certain ovarian characteristics, demonstrating that DHT-induced sex-ambiguous gonads were incomplete ovary-to-testis reversals (IOTTRs). The expression levels of some ovary-biased genes in the IOTTRs were significantly higher than in the control testes but lower than in the control ovaries. These results show that low concentrations of DHT induced complete or incomplete female-to-male sex reversal in P. nigromaculatus, and incomplete sex reversal retained certain ovarian characteristics not only at gonadal morphological and histological levels but also at the molecular level. They present study highlights potential risks of DHT and other androgenic EDCs for P. nigromaculatus. Environ Toxicol Chem 2015;34:2370–2377. © 2015 SETAC

The black-spotted pond frog Pelophylax nigromaculatus is a typical temperate-adapted amphibian species, largely endemic to East Asia. It occurs from central, east, and northeast China, to the Russian Far East, the Korean Peninsula, and part of Japan. So far, little is known about this frog from the Loess Plateau, a region that has experienced profound changes in climate and environment since the late Miocene. Particularly, between the Loess Plateau and the North China Plain, stand the Luliang and Taihang mountain ranges, which form an important geographical divide in East China. Therefore, frogs from the Loess Plateau are indispensable for understanding the biogeography and evolution of this species. In this study, a total of 933 specimens of P. nigromaculatus collected range-wide were sequenced for the mitochondrial Cyt b gene, with 345 of them being genotyped at 10 nuclear microsatellite loci. Both mitochondrial and microsatellite data resolved four major frog clades: (i) Loess Plateau clade, containing only frogs from the Loess Plateau; (ii) East China clade, containing frogs from the rest of China except those from the Liaodong Peninsula; (iii) Peninsulas clade, comprising frogs from the Korean Peninsula and Liaodong Peninsula; and (iv) Japan clade. Frogs from the Loess Plateau have deeply diverged from frogs in the remaining regions, with 11.5%–13.1% sequence divergence, which is comparable to distances between congeneric ranid species. Ecological niche modeling analysis showed that the potential distribution ranges of the Loess Plateau frogs and the non-Loess Plateau frogs are rather different; niche identity testing indicated that the environmental niches of frogs from the Loess Plateau and other regions are not equivalent. Our data thus suggest that the Loess Plateau frogs represent a cryptic species. The uplift of the Luliang Mountains was the most plausible trigger of this cryptic diversification.

Understanding the factors that contribute to loss of genetic diversity in fragmented populations is crucial for conservation measurements. Land-bridge archipelagoes offer ideal model systems for identifying the long-term effects of these factors on genetic variations in wild populations. In this study, we used nine microsatellite markers to quantify genetic diversity and differentiation of 810 pond frogs (Pelophylax nigromaculatus) from 24 islands of the Zhoushan Archipelago and three sites on nearby mainland China and estimated the effects of the island area, population size, time since island isolation, distance to the mainland and distance to the nearest larger island on reduced genetic diversity of insular populations. The mainland populations displayed higher genetic diversity than insular populations. Genetic differentiations and no obvious gene flow were detected among the frog populations on the islands. Hierarchical partitioning analysis showed that only time since island isolation (square-root-transformed) and population size (log-transformed) significantly contributed to insular genetic diversity. These results suggest that decreased genetic diversity and genetic differentiations among insular populations may have been caused by random genetic drift following isolation by rising sea levels during the Holocene. The results provide strong evidence for a relationship between retained genetic diversity and population size and time since island isolation for pond frogs on the islands, consistent with the prediction of the neutral theory for finite populations. Our study highlights the importance of the size and estimated isolation time of populations in understanding the mechanisms of genetic diversity loss and differentiation in fragmented wild populations.

Amphibians are globally threatened, but not all species are affected equally by different threatening processes. This is true for the threat posed by the chytridiomycete fungus (Batrachochytrium dendrobatidis). We compiled a European data set for B. dendrobatidis to analyze the trends of infection in European amphibians. The risk of infection was not randomly distributed geographically or taxonomically across Europe. Within countries with different prevalence, infection was nonrandom in certain amphibian taxa. Brown frogs of the genus Rana were unlikely to be infected, whereas frogs in the families Alytidae and Bombinatoridae were significantly more likely to be infected than predicted by chance. Frogs in the 2 families susceptible to B. dendrobatidis should form the core of attempts to develop spatial surveillance studies of chytridiomycosis in Europe. Ideally, surveys for B. dendrobatidis should be augmented by sampling the widespread genus Pelophylax because this taxon exhibits geographically inconsistent overinfection with B. dendrobatidis and surveillance of it may facilitate recognition of factors causing spatial variability of infection intensity. Several European amphibian taxa were not represented in our data set; however, surveillance of unsampled species should also occur when warranted. Resumen Los anfibios estan amenazados globalmente, pero los diferentes procesos de amenaza no afectan a todas las especies de la misma manera. Esto es cierto para la amenaza que representa el hongo quitridiomiceto (Batrachochytrium dendrobatidis). Compilamos un conjunto de datos europeos de B. dendrobatidis para analizar las tendencias de la infeccion en anfibios de Europa. La distribucion geografica o taxonomica del riesgo de infeccion no se distribuyo aleatoriamente en Europa. En paises con prevalencia diferente, la infeccion no fue aleatoria en ciertos taxa de anfibios. La infeccion fue poco probable en ranas del genero Rana, mientras que en ranas de las familias Alytidae y Bombinatoridae la probabilidad de infeccion fue significativamente mayor. Ranas de las 2 familias susceptibles a B. dendrobatidis deberian formar el nucleo de intentos por desarrollar estudios de vigilancia espacial de la quitridiomicosis en Europa. Idealmente, se deben incrementar los monitoreos de B. dendrobatidis mediante muestreos del genero Pelohylax por su distribucion amplia y porque presenta una sobreinfeccion con B. dendrobatidis geograficamente inconsistente y su monitoreo puede facilitar el reconocimiento de factores que produce variacion espacial de la intensidad de infeccion. Sin embargo, varios taxa de anfibios europeos no estaban representados en nuestro conjunto de datos y el monitoreo de especies no muestreadas tambien debe ocurrir cuando sea posible.

Summary The evolutionary potential and ecological importance of interspecific hybrids continues to be a controversial issue. Traditionally, hybridization – often associated with polyploidy and clonal reproduction – was considered an important mechanism for speciation in plants, but not in animals. More recently, investigations have shifted to the question: Under which genetic and ecological conditions do hybrid taxa and different ploidies arise and succeed, and when and where do they fail? Finding answers to this question is aggravated by the fact that suitable taxa for such studies are often far apart on the phylogenetic tree. Hence, results are influenced by many confounding variables. In this study, we reduce this problem by investigating the fitness within a complex of three closely related water frog taxa consisting of the two sexually reproducing parental species Pelophylax lessonae (genotype LL) and P. ridibundus (RR) plus their interspecific hybrid P. esculentus which comes in three ploidy types (LR, LLR and LRR), as well as with sexual and hemiclonal reproduction. Offspring of all five genotypes were produced by artificially crossing adults sampled from populations in Slovakia, Germany and Switzerland. This created genetic variation. They were then raised at two temperature levels: 18 and 24 °C. This created ecological variation. Larval performance under the two temperature regimes was analysed with respect to three fitness-related parameters: survival rate, days to metamorphosis and weight at tail resorption. Survival rate was significantly higher for offspring of the three hybrid types (LR, LLR and LRR) compared with those of the parental species (LL, RR), at both rearing temperatures. For days to metamorphosis and weight at metamorphosis, we found an interaction between offspring type and temperature. In both cases, performance of hybrid and parental offspring did not differ at 24 °C, but at 18 °C hybrids metamorphosed faster and at a lower weight than parentals. We discuss these results in relation to those from other studies and conclude that under cold conditions hybrids (especially the two triploid types) have higher fitness than both parental species. This genotype × environment interaction could be one reason why all-hybrid populations mainly occur at the cooler northern range of the water frog distribution.

SUMMARY 1. Although the influence of water availability and precipitation regimes on amphibians has been studied at large scales, whether and how interannual rainfall and hydrological variations affect amphibians dynamics at a local scale have rarely been addressed. In this respect, accounting for variations in species detectability in space and time has also been overlooked. 2. We assessed the effects of rainfall and hydrological variations on the breeding dynamics of three amphibian taxa: Pelodytes punctatus, Hyla meridionalis and Pelophylax spp. in 20 ponds of the Camargue region (southern France) over a 7-year study period. 3. We used multiple season occupancy models to test the effect of winter‐spring rainfall and interannual variations in hydroperiod, mean water depth and drought events on tadpole presence in spring (March‐June), a proxy for breeding dynamics. 4. We used an independent survey with spatial replicates (dipnet sweeps) to disentangle the relative contributions of phenology and detectability to the absence of records in a given month. For the three taxa considered, the probability of missing a species when that species was actually present in a pond was most often negligible. Hence, we could consider that multiseason models properly tracked changes in species phenology. 5. Pelodytes punctatus was first detected in March, while the two other taxa appeared later in April. Hyla meridionalis appeared as a mid-season species with much more synchronous pond occupancy than Pelodytes punctatus. The detection peak of Pelophylax spp. was short and unexpectedly early for this taxon. 6. Seasonal winter‐spring rainfall was associated with a decrease in extinction rates and even more strongly with an increase in colonisation rates at individual ponds. 7. Colonisation rate increased following an annual drought and was best modelled as a negative quadratic effect of the variance of pond hydroperiod. Extinction probability was best modelled by a negative quadratic effect of mean water level. Hence, breeding was more stochastic (i) in unpredictable and shallow ponds because of yearly drying up and (ii) in highly predictable and deep ponds, possibly due to the presence of predators such as fish and crayfish. 8. Overall, we show that ponds with intermediate rather than extreme variations in environmental conditions currently correspond to optimal breeding sites. Our study demonstrates that amphibian monitoring coupled with fine-scale analysis of environmental conditions is necessary to understand species dynamics in the long run and to inform conservation efforts for these species.

Water frogs inhabiting Cyprus represent a distinct evolutionary species of Messinian origin that is formally described in this paper. The systematic status of Cypriot frogs is evidenced by specific characters in their mitochondrial (mt) and nuclear (nu) DNA sequences, and the fact that they form a well supported monophyletic clade in both mtDNA and nuDNA phylogenies. While genetic data revealed clear and reproducible differences between this new taxon and all other western Palearctic water frog species including Pelophylax bedriagae in the Levant and two Anatolian water frogs lineages (P. cf. bedriagae -1 and P. cf. bedriagae -2), there is no diagnostic morphological or morphometric character that allows a clear discrimination between Cyprus frogs and frogs from the adjacent mainland. If several morphometric indices are combined as predictor variables in a discriminant analysis, however, both females and males of Cypriot water frogs are correctly distinguished from the other eastern Mediterranean lineages. While phylogenies based on concatenated sequences of two mitochondrial genes (ND2 + ND3) suggest a sister group relationship of Cypriot and Anatolian water frog lineages, our nuclear data hypothesize a sister group relationship between Cypriot frogs (sp. n.) and Crete frogs (P. cretensis), thus speaking for the same isolation time of both island populations (© 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim)

Western Palearctic water frogs in the genus Pelophylax are a set of morphologically similar anuran species that form hybridogenetic complexes. Fully reliable identification of species and especially of hybrid ploidy depends on karyological and molecular methods. In central Europe, native water frog populations consist of the Pelophylax esculentus complex, that is, P. lessonae (LL), P. ridibundus (RR) and the hybrid form P. esculentus that can have different karyotypes (RL, LLR and RRL). We developed existing molecular methods further and propose a simple PCR method based on size-differences in the length of the serum albumin intron-1 and the RanaCR1, a non-LTR retrotransposon of the chicken repeat (CR) family. This PCR yields taxon-specific banding patterns that can easily be screened by standard agarose gel electrophoresis and correctly identify species in all of the 160 samples that had been identified to karyotype with other methods. To distinguish ploidy levels in LR, LLR and RRL specimens, we used the ratio of the peak heights of the larger (ridibundus specific) to the smaller (lessonae specific) bands of fluorescently labelled PCR products resolved on a capillary DNA sequencer and obtained a correct assignment of the karyotype in 93% of cases. Our new method will cut down time and expenses drastically for a reliable identification of water frogs of the P. esculentus complex and potentially for identification of other hybridogenetic complexes and/or taxa, and it even serves as a good indicator of the ploidy status of hybrid individuals.

Aim To infer the evolutionary history of Rana (Pelophylax) lessonae Camerano within its inferred Quaternary refugial range, and to shed light on the processes that have contributed to shaping the patterns of diversity within the southern European peninsulas. Location The Italian peninsula south of the Alps and Sicily. Methods Sequence analysis of a mitochondrial cytochrome b gene fragment in 149 individuals sampled from 25 localities. Results Three mitochondrial DNA (mtDNA) phylogroups were identified, distributed in northern Italy, the whole Italian peninsula south of the northern Apennines, and Sicily. Syntopy between the northern and peninsular lineages was observed close to the northern Apennines. The northern lineage was the most differentiated, showing a net sequence divergence of 4.8 ± 0.8% with respect to the two others, whereas the net divergence between peninsular and Sicilian lineages was 2.6 ± 0.6%. Analysis of molecular variance (amova) revealed that 93% of the overall variation occurred between these three groups. Historical demographic statistics support a recent expansion for both the northern and peninsular groups, but not for the Sicilian group. In both northern and peninsular Italy, such an expansion was likely to have occurred during the last glaciation. Main conclusions Our results suggest that a number of microevolutionary processes were involved in shaping the present genetic structure of R. lessonae in Italy. These encompass allopatric differentiations in three distinct Pleistocene refugia, recent population expansions and secondary contacts. Our results, together with some previous work, support (1) the existence of a suture zone in the northern Apennines, and (2) the possibility of population expansions during the last glacial phase, when a vast widening of the lowland floodplain habitats followed sea-level fall, particularly in northern Italy. When compared with previous analyses of allozyme data, it appears that the peninsular mtDNA lineage has recently replaced the Sicilian one in southern Calabria, and we suggest that this event occurred due to selective introgression. The implications of such an occurrence for the study of factors underlying the patterns of diversity within this southern European biodiversity hotspot are discussed. Taxonomic implications of the results are also evaluated.

The rate of somatic development of anuran amphibians is only roughly correlated with the rate of gonad differentiation and varies among species. The somatic stage of a tadpole often does not reflect its age, which seems to be crucial for gonad differentiation rate. We compared the morphology and differentiation of developing ovaries at the light and electron microscopy level, with reference to somatic growth and age of a female. Our observations were performed on 12 species of six families (Rana lessonae, R. ridibunda, R. temporaria, R. arvalis, R. pipiens, R. catesbeiana, Bombina bombina, Hyla arborea, Bufo bufo. B. viridis, Xenopus laevis, Pelobates fuscus) and compared with the results obtained by other authors. This allowed us to describe the unified pattern of anuran female gonad differentiation. Ovary differentiation was divided into 10 stages: I-III, undifferentiated gonad; IV, sexual differentiation; V, first nests of meiocytes; VI, first diplotene oocytes; VII-IX, increasing number of diplotene oocytes and decreasing number of oogonia and nests; X, fully developed ovary composed of diplotene oocytes with rudimental patches of oogonia. We distinguished three types of ovary differentiation rate: basic (most species), retarded (genus Bufo), and accelerated (green frogs of the subgenus Pelophylax genus Rana).