It is well known that female adults of frog-biting midges (Corethrellidae) are attracted by anuran mating calls. However, the role of host-emitted cues other than mating calls in determining the level of attraction remains poorly understood. Here, we performed field playback experiments to test how the host-seeking behaviour of Corethrella Coquillett is influenced by different sound stimuli: mating call of a tree frog, cricket call, and no sound. Then, after long-distance host detection, we evaluated if Corethrella can distinguish landing sites by using at short distance the colour and/or shape of anurans. For that, we placed artificial models of frogs with different colours (light green and dark brown) and an artificial caterpillar model (light green colour) on the top of speakers. We found that only frog mating calls presented with the visual cue of an artificial frog were attacked by Corethrella, suggesting that frog shape, regardless of colour, is an important cue for the identification of its hosts. This is the first study providing evidence that after flying long distances guided by frog calls, Corethrella can identify their hosts using visual cues at short distances.

Studies integrating variation in species composition among sites are useful in understanding the impacts of land-use changes on the spatial distribution of biodiversity. However, the failure to recognize the distinction between beta diversity components, dissimilarity due to species replacement (i.e. turnover) and dissimilarity due to species loss from site to site (i.e. nestedness), can lead to inappropriate use of some indices. Here, we evaluated how the spatial distribution of anuran beta diversity components, turnover and nestedness, is associated with local and landscape descriptors in a tropical agricultural landscape with a recent history of agriculture expansion in south-eastern Brazil. Overall, 27 anuran species were found in the region with average ± SD species richness in each pool of 9.5 ± 3.5 species, ranging from 4 to 15 species. We observed that species turnover was the major component for anuran dissimilarity among pools, indicating that anuran species occurring in species-poor pools are not subsets of anuran species occurring in species-rich pools. Local variables and geographic distance were not important descriptors explaining the variation of anuran beta diversity. In contrast, the distance of the pools to the nearest forest fragment explained 16% of the variance in total beta diversity, 5% of the nestedness component and 2% of spatial turnover. Our results show that pools distributed across farmland landscapes are harbouring different anuran species composition, and together, these pools are contributing to the regional diversity of anurans in this region which is considered one of the most deforested and fragmented within Brazil.

Pathogens are increasingly implicated in amphibian declines but less is known about parasites and the role they play. We focused on a genus of nematodes (Rhabdias) that is widespread in amphibians and examined their genetic diversity, abundance (prevalence and intensity), and impact in a common toad (Rhinella horribilis) in Panama. Our molecular data show that toads were infected by at least four lineages of Rhabdias, most likely Rhabdias pseudosphaerocephala, and multiple lineages were present in the same geographic locality, the same host and even the same lung. Mean prevalence of infection per site was 63% and mean intensity of infection was 31 worms. There was a significant effect of host size on infection status in the wild: larger toads were more likely to be infected than were smaller conspecifics. Our experimental infections showed that toadlets that were penetrated by many infective Rhabdias larvae grew less than those who were penetrated by few larvae. Exposure to Rhabdias reduced toadlet locomotor performance (both sustained speed and endurance) but did not influence toadlet survival. The effects of Rhabdias infection on their host appear to be primarily sublethal, however, dose-dependent reduction in growth and an overall impaired locomotor performance still represents a significant reduction in host fitness.

Correlations between host phenotype and vulnerability to parasites can clarify the processes that enhance rates of parasitism, and the effects of parasites on their hosts. We studied an invasive parasite (the pentastome Raillietiella frenatus, subclass Pentastomida, order Cephalobaenida) infecting a new host (the invasive cane toad Rhinella marina), in tropical Australia. We dissected toads over a 27-month period to investigate seasonal changes in pentastome population dynamics and establish which aspects of host phenotype are related to infection. Pentastome prevalence and intensity varied seasonally; male toads were 4 times more likely to be infected than were females; and prevalence was highest in hosts of intermediate body size. The strong sex effect may reflect habitat or dietary divergence between the sexes, resulting in males encountering parasites more often. The relationship between pentastome prevalence and host size likely reflects a role for acquired immunity in preventing re-infection. Infection did not influence host body condition (fatbody size), suggesting that R. frenatus does not impose high energy costs in cane toads. Infected toads had heavier spleens (likely an immune response to infection) and larger testes (perhaps since reproductively active hosts have altered microhabitat use and/or immunocompetence) than did uninfected conspecifics. Although experimental studies are required to identify the causal bases of such patterns, our data confirm that infection status within a population can be strongly linked to host phenotypic traits.

Cane toads (Bufo marinus) were introduced to Australia in 1935 and have since spread widely over the continent, generating concern regarding ecological impacts on native predators. Most Australian cane toad populations are infected with lung nematodes Rhabdias pseudosphaerocephala, a parasite endemic to New World (native-range) cane toad populations; presumably introduced to Australia with its toad host. Considering the high intensities and prevalence reached by this parasite in Australian toad populations, and public ardour for developing a control plan for the invasive host species, the lack of experimental studies on this host-parasite system is surprising. To investigate the extent to which this lungworm influences cane toad viability, we experimentally infected metamorph toads (the smallest and presumably most vulnerable terrestrial phase of the anuran life cycle) with the helminth. Infected toads exhibited reduced survival and growth rates, impaired locomotor performance (both speed and endurance), and reduced prey intake. In summary, R. pseudosphaerocephala can substantially reduce the viability of metamorph cane toads.

Immediately after their transition from aquatic to terrestrial life, juveniles of many anuran species are restricted to the margins of natal ponds. Understanding the factors determining the duration of that pondside aggregation has direct management ramifications in the case of the invasive cane toad (Bufo marinus) in tropical Australia. Previous work suggests that dispersal confers biotic advantages (reduced risk of cannibalism, enhanced feeding opportunities) to juvenile toads, but desiccation risk constrains these small animals to the moist margins of the pond. If so, juvenile dispersal should be sensitive to fluctuating hydric conditions on a diel and seasonal cycle. We tested this prediction with field observations (monitoring of dispersal to and from the pond) and field experiments (manipulating hydric regimes). Our results support a dynamic model of juvenile distribution, with a primary role for temporal variations in desiccation risk as the primary factor driving dispersal. During the dry season, strong diel cycles in desiccation risk generate a ‘tidal’ flow of juveniles, dispersing out in the moist morning but retreating to the pond margins at midday. Dispersal rates were enhanced by artificial watering during the dry season, and by the onset of the wet season.

The effects of intermittent feeding on the growth of oophagous Chirixalus eiffingeri and herbivorous Chirixalus idiootocus tadpoles were studied. Tadpoles were fed, once every 1, 2, 4, 8, 14, or 20 days (feeding treatments 1D, 2D, 4D, 8D, 14D, and 20D, respectively). At each feeding, C. eiffingeri were given yolk from chicken eggs, and those of C. idiootocus were given boiled spinach, ad libitum. For each species, each treatment consisted of 12 replicates of one tadpole each. For C. eiffingeri, the 1D feeding treatment was eliminated because the tadpoles were unwilling to eat daily. Chirixalus eiffingeri tadpoles in the 2D and 4D feeding treatments grew fastest, while tadpoles in the 20D treatment grew slowest. At least five tadpoles from each treatment, except 20D treatment, reached metamorphosis. Chirixalus idiootocus tadpoles in the 1D and 2D feeding treatments showed signs of growing, but tadpoles in the other four treatments did not grow at all. Only tadpoles in the 1D treatment reached metamorphosis. In another experiment, yolk digestion in the stomach of C. eiffingeri tadpoles was monitored. Yolk use was slow and egg yolk remained in the stomach for at least 8 days. Chirixalus idiootocus tadpoles could not tolerate long periods without food, probably because their digestive tract had a short clearance time and their stomach was incapable of long-term food storage. In contrast, between feedings, C. eiffingeri tadpoles seemed to mobilize food stored in their stomach, providing a continuous supply of nutrition to facilitate normal growth and development. These results and observations provide important insights into the maternal brood care of female C. eiffingeri.

Vocalizations of Boophis madagascariensis (Rhacophoridae) males were recorded in a mid-elevation rain forest in eastern Madagascar. Call notes made by males of this species were classified into 28 types. This represents the largest known call repertoire of any amphibian. The calls range widely in spectral characteristics from a narrowband, nearly pure-tone to broadband ‘rip’ notes, to even broader band, pulsatile ‘iambic’ notes consisting of up to 23 pulses. Even considering all ‘iambic’ notes as variants of one type, there remain eight distinct call notes produced by this species. Morphological analysis of the vocal musculature of B. madagascariensis revealed no obvious adaptations for unusual glottal flexibility or function. Playback studies using a subset of the call notes as stimuli were carried out in the animals' natural habitat to determine the functional significance of several of the call notes. At the lowest playback levels, there was a significant tendency for males to produce rip notes in immediate response to either rip or iambic note stimuli. At the higher playback levels, we identified one sequence of iambic notes that consistently evoked a significant iambic note response from males. A permutation test on call note transitions during spontaneous calling revealed that call note pairs occurred inconsistently and unpredictably. The evolutionary significance of an extended vocal repertoire, matched counter-calling and a random call note production scheme is discussed.

Immunohistochemistry for choline acetyltransferase (ChAT) revealed an extensive network of cholinergic fibers in the tectum of amphibians. The distribution of ChAT immunoreactive fibers was not restricted to superficial retinocipient layers, but also included deep tectal layers. The aim of the present study was to investigate the origin of the cholinergic inputs to the tectum of amphibians. For that purpose, application of retrograde tracers in the tectum of the anuran Rana perezi and the urodele Pleurodeles waltl was combined with ChAT immunohistochemistry. Double-labeled cells were found primarily in the nucleus isthmi of both species. The cholinergic isthmotectal projection is bilateral and topographically arranged and all retrogradely labeled cells found in this nucleus were ChAT immunoreactive. Remarkably, abundant cholinergic cells in two tegmental nuclei, the pedunculopontine tegmental nucleus (anurans) and the laterodorsal tegmental nucleus (anurans and urodeles), were demonstrated to provide additional cholinergic innervation to the tectum. We compare the present results with previously reported studies in amphibians and other vertebrates, and discuss the possible functional significance of the cholinergic innervation of the amphibian tectum.