Influence of temperature on evaporative water loss and cutaneous resistance to water vapour diffusion in the orange-thighed frog (Litoria xanthomera)

We evaluated the effect of ambient temperatures between 25 and 43°C on the rate of evaporative water loss (EWL) in eight adult Litoria xanthomera (average body mass = 7.3 ± 0.6 g). Frogs were placed in a cylindrical chamber that permitted them to fully conceal their ventral surfaces using a water-conserving posture. Their EWL was 7.1 ± 0.7 mg g^–1 h^–1 at 25°C and reached 28.0 ± 2.5 mg g^–1 h^–1 at 43°C. Agar replicas of the frogs were used to evaluate boundary-layer resistances associated with the EWL measurements and, thus, to permit evaluation of cutaneous resistance to vapour diffusion (r_c) in live frogs. The r_c of L. xanthomera was stable over the temperature range of 25–35°C, averaging about 28 s cm^–1, and then declined stepwise with ambient temperatures above 37°C. The highest r_c recorded for each individual over the range of temperatures studied averaged 32.0 ± 1.2 s cm^–1. The thermolabile nature of r_c demonstrates a well developed thermoregulatory control of EWL in this species, a trait very similar in pattern and extent to that previously measured in the closely related Litoria chloris.

Spatial patterns of a southern bell frog litoria raniformis population in an agricultural landscape

We investigated changes in the spatial organization of individuals within a population of endangered Southern Bell Frogs Litoria raniformis over an eight-month period. Our results identified strong temporal changes in both spatial organization and the apparent location of L. raniformis within the study site. Ripley's K Function analyses showed that the position of individuals relative to one another shifted from random immediately after the study site was flooded (p < 0.005), to strongly clustered at spatial scales between 0-1 500 m during the peak breeding period (p < 0.005). The majority of flooded areas were dry by April and May and individuals again became aggregated within the remaining waterbodies.

Polymorphic microsatellite loci in the Australian tree frog, Litoria peronii

Eleven novel polymorphic microsatellite loci developed from a microsatellite enriched genomic library, are presented for the Australian tree frog <i>Litoria peroniii>. We screened 29 individuals from a single population and detected high levels of polymorphism for all 11 loci with the number of alleles/locus ranging from 9 to 24. Values of expected and observed heterozygosities ranged from 0.789–0.955 and 0.207–1.00, respectively. These microsatellite markers should prove useful in determining levels of genetic diversity, measuring gene flow and migration, assigning individuals to their most likely population of origin, and in the assignment of paternity.

Within-population variation in ejaculate characteristics in a prolonged breeder, Peron's tree frog, Litoria peronii

Sperm number is often a good predictor of success in sperm competition; however, it has become increasingly clear that, for some species, variation in probability of paternity cannot be explained by sperm number alone. Intraspecific variation in ejaculate characteristics, such as the number of viable sperm and sperm longevity, may play an equally important role in determining fertilization success. Here, we assess variation among ejaculates in three factors that may contribute to fertilization success (number of sperm per ejaculate, viability, and longevity), in a population of Peron’s tree frog (Litoria peronii). We detected large variation among males in the number of sperm per ejaculate and the proportion of viable sperm within ejaculates, which could not be explained by variation in either male size or body condition. However, the proportion of viable sperm released by males increased over the season. Finally, we assessed sperm longevity (proportion viable sperm determined using a dual-fluorochrome vital dye) at two different temperatures. At 23°C, on average, 75% of sperm remained viable after 2 h, but there were significant differences amongst males with the percentage of viable sperm ranging from 43% to 95%. For sperm incubated at 4°C, ejaculates varied fivefold in sperm longevity with some males having 50% viable sperm after 5 days. Our data suggest that ejaculate characteristics (sperm number, viability, and longevity) vary widely in Peron’s tree frog and may therefore play an important role in determining siring success both in the presence and absence of sperm competition. We discuss the results in relation to selection on ejaculate traits via natural and sexual selection in this and other amphibians.

Consistent paternity skew through ontogeny in Peron's tree frog (Litoria peronii)

Background
A large number of studies in postcopulatory sexual selection use paternity success as a proxy for fertilization success. However, selective mortality during embryonic development can lead to skews in paternity in situations of polyandry and sperm competition. Thus, when assessment of paternity fails to incorporate mortality skews during early ontogeny, this may interfere with correct interpretation of results and subsequent evolutionary inference. In a previous series of in vitro sperm competition experiments with amphibians (Litoria peronii), we showed skewed paternity patterns towards males more genetically similar to the female.

Methodology/Principal Findings
Here we use in vitro fertilizations and sperm competition trials to test if this pattern of paternity of fully developed tadpoles reflects patterns of paternity at fertilization and if paternity skews changes during embryonic development. We show that there is no selective mortality through ontogeny and that patterns of paternity of hatched tadpoles reflects success of competing males in sperm competition at fertilization.

Conclusions/Significance
While this study shows that previous inferences of fertilization success from paternity data are valid for this species, rigorous testing of these assumptions is required to ensure that differential embryonic mortality does not confound estimations of true fertilization success.

Male and female effects on fertilization success and offspring viability in the Peron's tree frog, Litoria peronii

There is increasing theoretical and empirical evidence that genetic compatibility among partners is an important determinant of fertilization success and offspring viability. In amphibians, females often actively choose partners from among a variety of males and polyandry is common. Genetic compatibility among partners may therefore be an important determinant of fertilization success and offspring viability in some amphibians. Amphibians also show some of the highest levels of genetic differentiation among neighbouring populations known in vertebrates, and as such, populations may have evolved different co-adapted gene complexes. This means that offspring from among-population crosses may have reduced fitness. It is therefore essential to understand to what extent crossings between and within populations may interfere with successful fertilization and offspring viability. Here, we test whether crossing individuals within and between two different populations of the Australian Peron's tree frog (Litoria peronii) using artificial fertilizations affect fertilization success and offspring viability. Fertilization success per se is strongly influenced by male identity, which is likely to depend at least to some extent on the experimental procedure (e.g. resulting in variation in sperm number per ejaculate), whereas there was no fertilization effect of female identity. More importantly, male and female identity, independently of each other, explained significant variation in offspring viability, whereas no such effect could be linked to population of origin. Thus, our experiments suggest that crossing populations may not always be the most significant factor affecting fertilization success or offspring viability, but may be more influenced by the genetic quality or the genetic compatibility of partners.

Female choice for males with greater fertilization success in the Swedish Moor Frog, Rana arvalis

Background: Studies of mate choice in anuran amphibians have shown female preference for a wide range of male traits despite females gaining no direct resources from males (i.e. non-resource based mating system). Nevertheless, theoretical and empirical studies have shown that females may still gain indirect genetic benefits from choosing males of higher genetic quality and thereby increase their reproductive success.
Methodology/Principal Findings: We investigated two components of sexual selection in the Moor frog (Rana arvalis), precopulatory female choice between two males of different size (‘large’ vs. ‘small’), and their fertilization success in sperm competition and in isolation. Females’ showed no significant preference for male size (13 small and six large male preferences) but associated preferentially with the male that subsequently was the most successful at fertilizing her eggs in isolation. Siring success of males in competitive fertilizations was unrelated to genetic similarity with the female and we detected no effect of sperm viability on fertilization success. There was, however, a strong positive association between a male’s innate fertilization ability with a female and his siring success in sperm competition. We also detected a strong negative effect of a male’s thumb length on his competitive siring success.
Conclusions/Significance: Our results show that females show no preference for male size but are still able to choose males which have greater fertilization success. Genetic similarity and differences in the proportion of viable sperm within a males ejaculate do not appear to affect siring success. These results could be explained through pre- and/or postcopulatory choice for genetic benefits and suggest that females are able to perceive the genetic quality of males, possibly basing their choice on multiple phenotypic male traits.

Characterization of nine polymorphic microsatellite loci in the dyeing poison frog Dendrobates tinctorius (Dendrobatidae), and their cross-species utility in two other dendrobatoid species

While field and laboratory based studies have provided significant insights into the parental care and courtship behaviour of dendrobatoid frogs, a comprehensive assessment of their genetic mating systems and population genetic parameters has been precluded because ofthe lack of highly variable DNA markers. Here we document the development of nine novel polymorphic microsatellite markers for the dyeing poison frog Dendrobates tinct or ius (Dendrobatidae ). We found between three and 16 alleles per locus in 60 individuals (30 males, 30 females) from the field site Saut Parare, French Guiana, with an average observed heterozygosity of 0. 75. None of the loci deviated significantly from Hardy-Weinberg equilibrium or showed linkage disequilibrium. We also report successful cross-species amplification of the nine markers in two other dendrobatoid species (Allobates femora/is and Oophaga pumilio). These markers have the potential to aid in determining the genetic structure of local populations, identifying small-scale phylogenies such as parent-offspring relationships and will allow for cross-species comparisons within dendrobatoid species. Therefore, these markers can be applied to a wide range of scientific fields, such as conservation, behavioural ecology and evolutionary biology.

Noisy neighbours at the frog pond: effects of invasive cane toads on the calling behaviour of native Australian frogs

Invasive species can disrupt the communication systems that native biota use for reproductive interactions. In tropical Australia, invasive cane toads (Rhinella marina) breed in many of the same waterbodies that are used by native frogs, and males of both the invader and the native taxa rely on vocal signals to attract mates. We conducted playback experiments to test the hypothesis that calls of toads may influence the calling behaviour of frogs (Limnodynastes convexiusculus and Litoria rothii). Male L. convexiusculus adjusted their calling rate and the variance in inter-call interval in response to a variety of sounds, including the calls of cane toads as well as those of other native frog species, and other anthropogenic noise, whereas L. rothii did not. Within the stimulus periods of playbacks, male L. convexiusculus called more intensely during long silent gaps than during calling blocks. Thus, males of one frog species reduced their calling rate, possibly to minimise energy expenditure during periods of acoustic interference generated by cane toads. In spite of such modifications, the number of overlapping calls (within stimulus periods) did not differ significantly from that expected by chance. In natural conditions, the calls of cane toads are continuous rather than episodic, leaving fewer gaps of silence that male frogs could exploit. Future work could usefully quantify the magnitude of temporal (e.g. diel and seasonal) and spatial overlap between calling by toads and by frogs and the impact of call-structure shifts on the ability of male frogs to attract receptive females.

Low impact of chytridiomycosis on frog recruitment enables persistence in refuges despite high adult mortality

The global chytridiomycosis pandemic caused by the pathogen Batrachochytrium dendrobatidis (. Bd) is implicated in the apparent extinction or severe decline of over 200 amphibian species. Many declined species now only persist in isolated remnant populations. In this study we examine how remnant populations coexist with Bd, focusing on disease impact on adult survival and recruitment potential in the chytridiomycosis-threatened frog Litoria verreauxii alpina. Using skeletochronology we found that the adult male population in both 2011 and 2012 was dominated by a two year old age cohort. The lack of recruitment into the three year old cohort in 2012 indicates that annual adult survival is very low. Combined with high Bd prevalence and heavy infection burdens, the pathogen appears to drive almost complete mortality of breeding adults over their first breeding season. However, adults successfully mate prior to large increases in disease prevalence that occurs during the breeding season. Infection prevalence among tadpoles and juveniles is low. Exposure to warm water could provide a mechanism for avoiding or clearing Bd infection. Relatively low Bd prevalence in juveniles prior to dispersal into terrestrial habitat indicates that Bd has minimal impact on early life history stages. As such, recruitment is probably high, allowing populations to persist despite low adult survival. This dependence on reliable annual recruitment may explain why remnant populations persist in permanent ponds rather than ephemeral ponds that were historically occupied. New management strategies that focus on increasing recruitment may provide a way forward for the management of disease-threatened amphibian species.