The effects of time,space and spectrum on auditory grouping in túngara frogs

Male túngara frogs (Physalaemus pustulosus) produce complex calls consisting of two components, a ~350 ms FM sweep called the “whine” followed by up to seven ~40 ms harmonic bursts called “chucks”. In order to choose and locate a calling male, females attending to choruses must group call components into auditory streams to correctly assign calls to their sources. Previously we showed that spatial cues play a limited role in grouping: calls with normal spectra and temporal structure are grouped over wide angular separations (≤135°). In this study we again use phonotaxis to first test whether an alternative cue, the sequence of call components, plays a role in auditory grouping and second, whether grouping is mediated by peripheral or central mechanisms. We found that while grouping is not limited to the natural call sequence, it does vary with the relative onset times of the two calls. To test whether overlapping stimulation in the periphery is required for grouping, the whine and chuck were filtered to restrict their spectra to the sensitivity ranges of the amphibian and basilar papillae, respectively. For these dichotic-like stimuli, grouping still occurred (albeit only to 45° separation), suggesting that stream formation is mediated by central mechanisms.     

Habitat associations of pool-breeding amphibians in eastern Massachusetts,USA

There is a pressing need to develop a sound conservation strategy for pool-breeding amphibians, which includes gaining a better understanding of the habitat and landscape-scale characteristics associated with populations. To investigate relationships between amphibian species richness and characteristics of breeding pools and surrounding landscapes, we surveyed 85 pools in eastern Massachusetts (USA) in 1996 and 1997. A total of 11 species was detected, with most pools having 2–5 species. Pools were typically small, 77.6% were <0.2 ha, but most pools (72%) had hydroperiods that persisted at least into August in most years. Based on linear regression analyses, species richness was positively associated with three within-pool variables (pool surface area, hydroperiod, and the amount of emergent vegetation), and a landscape-level variable (presence of another breeding pool within 1 km), while one within-pool variable (tree canopy cover) exhibited a significant negative association with species richness. These within-pool habitat variables and connectivity to other breeding pools are important characteristics to consider when attempting to identify breeding sites that could provide core habitat in conservation reserves designed for the conservation of pool-breeding amphibian species richness. Conservation reserves for pool-breeding amphibian populations should include pool complexes functioning as habitat for metapopulations. Core pool habitats within such reserves should be large (0.5–1.0 ha), with seasonal hydroperiods that persist into August or that dry in some years, and with sufficient emergent vegetation to provide diverse microhabitats and refugia.     

Conserving pool-breeding amphibians in human-dominated landscapes through local implementation of Best Development Practices

Seasonal forest pools in the northeastern USA are unique ecosystems whose functions are intimately associated with adjacent upland habitats. This connection, coupled with their small size and ephemeral surface water, has made conservation of pool resources challenging. Seasonal pools provide optimal breeding habitat for animals adapted to temporary waters including ambystomatid salamanders (Ambystoma spp.), wood frogs (Rana sylvatica LeConte), and some invertebrates and plants. To date, wetland conservation efforts have been primarily limited to 2 pathways: land use regulation and preservation. Although both of these pathways have the potential to conserve some pool resources, they are often insufficient to maintain an array of pools in the landscape that support local population dynamics of amphibians. We propose a third pathway – local land-use planning – that can complement regulatory and preservation efforts. This suite of strategies, embodied in our Best Development Practices (BDPs), recognizes that not all pools will be conserved; local governances will need to develop priorities for conservation. The BDPs encourage local governances to (1) proactively identify their pool resources, (2) rank those pools according to their relative ecological value, and (3) establish management procedures and apply recommended guidelines in accordance with the relative rankings. We recommend that pools be ranked using biological criteria (e.g., presence of listed species, presence of breeding species, and egg mass abundance) and on the availability and quality of adjacent terrestrial habitat. We recommend 3 management zones: the pool depression, the pool envelope (i.e., land within 30 m of the pool), and the critical terrestrial habitat (i.e., 30–230 m from the pool). Residential, industrial, and commercial development, which may compromise pool habitat (e.g., through building and road construction, site clearing, stormwater management, and lighting), should follow the recommended guidelines presented in Appendix 1 of this paper. Planning at the watershed level, using such tools as overlay zones, wetland ordinances, and easements, should lead to more effective, long-term management of, at a minimum, the most ecologically important seasonal forest pool resources and will provide developers with clear development guidelines. This process is already being successfully implemented in a number of New England towns.     

Effect of exogenous JHI on imaginal determination in Aeshna cyanea

Last instar larvae of Aeshna cyanea injected with 8 μg or more JHI before the end of the fourth ocular stage (day 7 or 8 of the instar) became perfect supernumerary larvae which indicates that adult determination does not occur before this time. When JHI is applied later (days 9–13) larval-adult intermediates are formed. It seems, therefore, that imaginal programming takes place at the end of the fourth ocular stage and might be correlated with the first ecdysone peak which occurs at the same time.     

Relationship between an increase of juvenile hormone titer in early instars and the induction of diapause in fully grown larvae of Sesamia nonagrioides

The larvae of Sesamia nonagrioides (Lepidoptera: Noctuidae) grown at 25 degrees C and long photoperiod (16:8h light:dark) pupate in the 5th or 6th (mostly) larval instar, while the larvae reared under a short photoperiod (12:12h) enter diapause during which they consume some food and undergo up to 12 (usually 3-4) stationary larval molts. Diapause programming includes an increase of juvenile hormone (JH) titer in the hemolymph from about 20 to 50 nM in the 4th and 5th instar larvae (titer in earlier instars was not measured). JH I, II, and III are present in approximate ratio 1-2:10:1. The JH titer drops to zero before pupation but remains around 20 nM during diapause. Perfect extra larval molts associated with a body weight increase can be induced in the non-diapausing larvae with a JH analogue (JHA). The weight rise is due to accumulation of reserves and not to a general body growth. The timing of extra molts is similar to the molting pattern of the diapausing larvae only when JHA is present since early larval instars. In the diapausing larvae, JHA application affects neither molting periodicity nor the body weight. It is concluded that (1) Increased JH titer in early larval instars is a part of diapause programming; (2) The extension of larval stage in the diapausing larvae, but not the timing pattern of extra molts, is due to continuously high JH titer; (3) The diapause program includes low food intake, maintenance of a certain body weight, and periodic larval molts.     

Cryopreservation of spermatozoa of the terrestrial Puerto Rican frog, Eleutherodactylus coqui

World-wide reports of amphibian population declines have led to increased interest in the reproductive biology of anurans. As a model system, here we present evidence for the effective cryoprotection of sperm from the Puerto Rican frog, Eleutherodactylus coqui, using mixtures of fetal bovine serum (FBS) and dimethylsulfoxide (Me(2)SO), glycerol or sucrose extenders. Using a fluorescent dye exclusion assay, we found that 53.9 and 50.4% of all sperm with intact membranes prior to freezing maintained membrane integrity after rapid freezing and thawing when protected with either a FBS/glycerol or FBS/sucrose solution, respectively. The methods reported here may be useful for similar work with many of the more than 700 other species in this genus.     

Chytridiomycosis impacts predator-prey interactions in larval amphibian communities

Despite ecologists increasingly recognizing pathogens as playing significant roles in community dynamics, few experimental studies have quantified patterns of disease impacts on natural systems. Amphibians are experiencing population declines, and a fungal pathogen (Batrachochytrium dendrobatidis; Chytridiomycota) is a suspected causal agent in many declines. We studied the effects of a pathogenic fungus on community interactions between the gray treefrog, Hyla chrysoscelis, and eastern newts, Notophthalmus viridescens. Recent studies have characterized chytridiomycosis as an emerging infectious disease, whose suspected rapid range expansion and widespread occurrence pose a significant risk for amphibian populations worldwide. We reared larvae in outdoor polyethylene experimental tanks and tested the effects of initial larval density, predator presence, and fungal exposure on Hyla recruitment and predator-prey interactions between Hyla and Notophthalmus. Newts reduced treefrog survival, and high intraspecific density decreased metamorphic body mass independent of B. dendrobatidis. The presence of fungi reduced treefrog body mass at metamorphosis by 34%, but had no significant main effect on survival or larval period length. B. dendrobatidis differentially affected larval development in the presence of predators; Hyla developed slower when reared with the pathogen, but only when newts were present. This significant predator-by-pathogen interaction suggests that the impact of chytridiomycosis on larval amphibians may be exacerbated in complex communities. Our data suggest that B. dendrobatidis effects on host life history may be complex and indirect. Direct measurements of the community-level effects of pathogens offer an important opportunity to understand a significant threat to global biodiversity—declining amphibian populations.     

Changes in RNA,DNA, protein contents and growth of turbot Scophthalmus maximus larvae and juveniles

The growth potential of turbot Scophthalmus maximus larvae and juveniles was studied using nucleic acid‐based indices and protein variables. The experiment was carried out from 4 to 60 days post hatching (dph). A significant increase in instantaneous growth rate during metamorphosis and retarded growth rate during post‐metamorphic phase were observed. Ontogenetic patterns of DNA, RNA and protein all showed developmental stage‐specific traits. The RNA:DNA ratio decreased up to 12 dph, then increased rapidly till 19 dph and fluctuated until 35 dph followed by a decline to the end. The RNA:DNA ratio was positively correlated with growth rate of juveniles during the post‐metamorphic phase, whereas this ratio was not a sensitive indicator of growth during the pre‐metamorphic phase and metamorphosis. The protein:DNA ratio showed a similar tendency to the RNA:DNA ratio. Changes of DNA content and protein:DNA ratio revealed that growth of S. maximus performed mainly by hyperplasia from 4 to 12 dph and hypertrophy until 21 dph during the pre‐metamorphic larval phase. Growth was dominantly hypertrophical from the early‐ to mid‐metamorphosing phase and hyperplastic thereafter. The results show that the DNA content and protein:DNA ratio can evaluate growth rates of larval and juvenile S. maximus on a cellular level.     

Auditory response characteristics of the piebald odorous frog and their implications

The piebald odorous frog (Odorrana schmackeri), the large odorous frog (Odorrana livida) and the concave-eared torrent frog (Amolops tormotus) are sympatric species living near the same torrent streams in the vicinity of Mt. Huangshan, China. A recent study demonstrated that A. tormotus can use sound signals involving ultrasonic components for communication in a noisy environment, and another sympatric species, O. livida, can also perceive ultrasonic sound. Here we report data on the hearing range of O. schmackeri by studying auditory evoked potentials and single-unit data from the torus semicircularis. This frog exhibits its two most sensitive peaks at 2 kHz and 3.5–4.0 kHz with thresholds <42 dB SPL, with an upper frequency limit of hearing at 8.5 kHz with threshold of 87 dB SPL. The upper limit is much lower than those of O. livida and A. tormotus, at 22 and 34 kHz, respectively. It suggests that sympatric species may respond differently to similar environmental selection pressures sculpting auditory communication systems.