External and internal control of patterns of feeding, growth and gonadal function in a temperate zone anuran, the toad Bufo bufo

Young toads were brought into the laboratory before hibernation in the autumn and exposed to alternating periods of low temperature and high temperature and feeding, thus simulating climatic cycles. In newly metamorphosed, non-hibernating toads, food intake was initially low and growth was stagnant, but subsequently food intake and lean growth increased strongly. Presumably, growth was initiated independently of feeding. In one-year-old, non-hibernating toads, food intake and growth declined with time, but high rates of feeding and growth were restored in the toads exposed to simulated hibernation. In both metamorphosed toads and one-year-old, hibernating toads, a period of predominant fat deposition succeeded predominant lean growth before food intake again declined to low levels. Initiation of sexual maturation in the female toads coincided with suspended growth, whereas sexual maturation in males was not correlated with changes in growth rate. In the female, but not the male toads, sexual maturation was greatly advanced under the experimental conditions, resulting in adult female sizes far below those typical of the wild population.     

Winter Hibernation and UCHL1-p34cdc2 Association in Toad Oocyte Maturation Competence

Currently, it is believed that toad oocyte maturation is dependent on the physiological conditions of winter hibernation. Previous antibody-blocking experiments have demonstrated that toad ubiquitin carboxyl-terminal hydrolase L1 (tUCHL1) is necessary for germinal vesicle breakdown during toad oocyte maturation. In this paper, we first supply evidence that tUCHL1 is highly evolutionarily conserved. Then, we exclude protein availability and ubiquitin carboxyl-terminal hydrolase enzyme activity as factors in the response of oocytes to winter hibernation. In the context of MPF (maturation promoting factor) controlling oocyte maturation and to further understand the role of UCHL1 in oocyte maturation, we performed adsorption and co-immunoprecipitation experiments using toad oocyte protein extracts and determined that tUCHL1 is associated with MPF in toad oocytes. Recombinant tUCHL1 absorbed p34^cdc2, a component of MPF, in obviously larger quantities from mature oocytes than from immature oocytes, and p13^suc1 was isolated from tUCHL1 with a dependence on the ATP regeneration system, suggesting that still other functions may be involved in their association that require phosphorylation. In oocytes from hibernation-interrupted toads, the p34^cdc2 protein level was significantly lower than in oocytes from toads in artificial hibernation, providing an explanation for the different quantities isolated by recombinant tUCHL1 pull-down and, more importantly, identifying a mechanism involved in the toad oocyte’s dependence on a low environmental temperature during winter hibernation. Therefore, in toads, tUCHL1 binds p34^cdc2 and plays a role in oocyte maturation. However, neither tUCHL1 nor cyclin B1 respond to low temperatures to facilitate oocyte maturation competence during winter hibernation.     

中华蟾蜍糖原含量的季节变化

于2005年11月至2006年10月,用硫酸-蒽酮比色法和比重法测定了中华蟾蜍各月份的肝糖原和肌糖原含量及肝比重。结果显示:冬眠期间(11月至次年的2月),糖原含量逐月下降;2月份时出现临时回升,然后继续下降;4月份时肝糖原含量最低;5月份起,肝糖原含量逐渐上升;5月份时肌糖原含量为最低;6月份起,肌糖原含量逐渐上升。虽然在7-8月间出现过下降,两种糖原的含量在10月份时达到一年中的最高值。这些结果表明,蟾蜍糖原含量在一年中呈现显著的季节性波动。越冬前所储备糖原的一部分可能用于越冬期间维持高水平的血糖,一部分用于基础代谢。2月份时糖原含量的临时上升,可能是血液中作为防冻保护剂的葡萄糖运回肝脏和肌肉中再合成糖原的结果。7-8月间糖原含量降低可能与蟾蜍夏蛰有关。雌性5月至10月期间的肝糖原总体水平显著低于雄性,可能与依赖可得到葡萄糖的卵母细胞中的糖原合成有关。糖原含量的季节变化与蟾蜍的生活状态(越冬、繁殖等)有关,并与血糖含量有联动关系。     

人工诱导冬眠对中华蟾蜍血液和组织中宏量营养素的影响

为探寻冬眠期间两栖动物血液和组织中宏量营养素的适应性改变过程,经人工诱导冬眠,检测了中华蟾蜍(Bufo gargarizans)在冬眠第1、3、7、14、28、42、56天的体重和脏器指数,以及血液、心、肝和骨骼肌组织中宏量营养素的含量。结果显示:1)冬眠期间中华蟾蜍体重未出现显著性变化,无性别差异。雄性蟾蜍的心、肝和腓肠肌的脏器指数显著性大于雌性(P0.01),但同一性别的脏器指数在冬眠期间无显著性变化。2)血中葡萄糖浓度自冬眠第42天起显著下降(P0.01);总蛋白在冬眠后第56天显著降低(P0.05),总胆固醇变化不显著。血中宏量营养素无性别差异。3)肝糖原自冬眠第42天起显著下降(P0.01),肌糖原自冬眠第1天起显著性下降(P0.05),而骨骼肌和心肌组织蛋白含量无显著变化。组织中宏量营养素无性别差异。人工诱导冬眠条件下,中华蟾蜍血液和组织中的糖类含量先迅速下降,血液中的蛋白成分只在深眠时才显著减少,但血液和组织中的宏量营养素水平可在1个月内维持稳定,这可能是其适应冬眠的主要生理学机制之一。     

Rapid expansion of the cane toad (Bufo marinus) invasion front in tropical Australia

Abstract Cane toads (Bufo marinus) are large toxic anurans that have spread through much of tropical Australia since their introduction in 1935. Our surveys of the location of the toad invasion front in 2001 to 2005, and radiotracking of toads at the front near Darwin in 2005, reveal much faster westwards expansion than was recorded in earlier stages of toad invasion through Queensland. Since reaching the wet‐dry tropics of the Northern Territory, the toads have progressed an average of approximately 55 km year⁻¹ (mean rate of advance 264 m night⁻¹ along a frequently monitored 55‐km road transect during the wet season of 2004–2005). Radiotracking suggests that this displacement is due to rapid locomotion by free‐ranging toads rather than human‐assisted dispersal; individual toads frequently moved >200 m in a single night. One radiotracked toad moved >21 800 m in a 30‐day period; the fastest rate of movement yet recorded for any anuran. Daily displacements of radiotracked toads varied with time and local weather conditions, and were highest early in the wet season on warm, wet and windy nights. The accelerated rate of expansion of the front may reflect either, or both: (i) evolved changes in toads or (ii) that toads have now entered an environment more favourable to spread. This accelerated rate of expansion means that toads will reach the Western Australian border and their maximal range in northern Australia sooner than previously predicted.     

Toads in the backyard: why do invasive cane toads (Rhinella marina) prefer buildings to bushland?

Like many invasive species, cane toads (Rhinella marina) in Australia concentrate in the disturbed habitats created by human activity, rather than in pristine areas. We surveyed cane toads in the wet–dry tropics of the Northern Territory to assess the abundances, body sizes, sexes, behaviour, hydration state and feeding rates of toads around buildings compared to those in areas remote from buildings, and conducted experimental trials to assess the effects of building-related variables (lights and increased toad densities) on the foraging success of toads. Toads around buildings were smaller than bushland conspecifics, and adult sex-ratios were female-biased. Toads were more sedentary around buildings than in the bush, but their feeding rates (based on direct observations and faeces production post-capture) were similar. That similarity, despite twofold-higher densities of competing toads around building, reflected the strong enhancement of feeding rates due to artificial lights attracting insects (in our experimental trials, a threefold increase regardless of the number of competing toads). Toads collected from around buildings were apparently in better hydric condition. Thus, access to water also may attract toads to buildings. The relative scarcity of adult male toads around buildings likely reflects waterbody-centred reproductive activities, whereas the concentration of females and juveniles around buildings is driven largely by access to the insects attracted by artificial light. We conclude that buildings enhance the persistence of cane toad populations and may facilitate their spread.     

Seasonal changes in the preferred body temperature, cardiovascular, and respiratory responses to hypoxia in the toad, Bufo paracnemis

Estivation is accompanied by a reduction of oxygen consumption in amphibians during drought. We tested the hypothesis that, during the dry season, the toad Bufo paracnemis selects a lower preferred body temperature (T(b)), and would be less sensitive to hypoxia, than during its active period. Therefore, during winter (dry season in S?o Paulo state, Brazil) and summer, we measured the effects of hypoxia (7% inspired O(2)) on preferred T(b). Additionally, pulmonary ventilation, heart rate, blood pressure, and oxygen consumption were also measured in toads at 15 and 25 degrees C. Blood gases were measured at 25 degrees C. Oxygen consumption was significantly higher during summer in toads at 25 degrees C. Under normoxia, preferred T(b) was higher during summer than during winter, and hypoxia caused a drop in preferred T(b) during both seasons. In both seasons, toads at 15 degrees C showed reduced pulmonary ventilation, heart rate, and blood pressure, and hypoxia had no effect. At 25 degrees C during summer only, hypoxia caused an increase in ventilation. Season had no effect on blood gases. We conclude that B. paracnemis displays an endogenous seasonal pattern of thermoregulation and control of ventilation. The decreased preferred T(b) and the physiological responses to hypoxia may be beneficial to toads encountering drought and when food is not available.     

Interspecific spawning between Common frogs (Rana temporaria) and Common toads (Bufo bufo)

The interaction between Common frogs (Rana temporaria) and Common toads (Bufo bufo) during the breeding season was studied at a small pond on Portland, Dorset. Although the frogs started and finished spawning earlier than the toads there was a period during which interspecific spawning between female toads and male frogs took place. This appears to have resulted from reduced male toad activity caused by the cold breeding season given that spatial, temporal and behavioural species separation did not occur.     

The spatial ecology of cane toads (Bufo marinus) in tropical Australia: Why do metamorph toads stay near the water?

Abstract Cane toads (Bufo marinus) have invaded large areas of Australia, killing many native predators as they have done so. The metamorph stage of the life cycle – the first terrestrial phase, immediately after transformation from the tadpole – is critical for ecological impact (because these animals are small enough to be prey for many native predators) and for potential control of toad populations (because small body size renders metamorphs vulnerable to desiccation). To quantify the spatial and temporal distribution of metamorph toads, and the biotic and abiotic factors that might affect their distributions, we surveyed toad breeding sites in Australia's wet‐dry tropics (Adelaide River floodplain, NT) in both the wet season and the dry season. Metamorphs were concentrated close to the water's edge during the dry season, especially at midday when desiccation rates were highest. During the wet season, metamorphs were widely dispersed through the landscape. Our surveys indicate that abiotic factors (risk of desiccation) are most favourable for metamorph toads close to the pond edge, but biotic factors (food supply, and risk of competition and cannibalism) are most favourable away from the water. Operative temperatures were spatially homogeneous and sublethal, and so are unlikely to influence metamorph distribution. Desiccation risk fluctuated on a diel cycle as well as seasonally. We predict that metamorph toads benefit from dispersing as soon as desiccation risk allows them to do so, and hence the distribution of metamorph toads will shift dynamically in response to weather‐mediated changes in rates of evaporative water loss.     

Effects of age, weight, hormones, and hibernation on breeding success in boreal toads (Bufo boreas boreas)

The goals of this study were to test the effects of exogenous hormones and hibernation on breeding behavior and gamete release by boreal toads (Bufo boreas boreas). Each year, a subset of 77 toads was hibernated and then paired with hibernated or nonhibernated mates and treated with luteinizing hormone releasing hormone analogue (LHRHa), human chorionic gonadotropin (hCG), or left untreated. Amplexus and egg and sperm production were recorded. At 1 yr of age, only 19% of pairs exhibited amplexus, and no sperm or eggs were produced. At 2 and 3 yr of age, most male toads treated with LHRHa exhibited amplexus (56.9% and 100%, respectively). Among 2-yr-old males, amplexus was more prevalent (P < 0.05) in those that were hibernated than in those that were nonhibernated (54.0% and 33.3%, respectively), but most males in each group (93.3% and 75%, respectively) produced sperm in response to LHRHa treatment. Only one 2-yr-old and two 3-yr-old females produced eggs. At 4 yr of age, eight females produced eggs, but two died from egg retention. More nonhibernated than hibernated females developed eggs (7 of 10 vs. 1 of 10, P < 0.05). Mean (±SD) weight of female toads producing eggs (58.9 ± 11.9 g) was greater (P < 0.05) than that of nonproducing females (43.6 ± 7.0 g). Similarly, four of seven nonhibernated females (58.8 ± 8.3 g) produced eggs at 5 yr of age. All eggs were produced by females treated once with LHRHa. Number of eggs per female varied (141 to 3307), and development to tadpoles was low (0 to 36.5%), although tadpoles did become toadlets. In conclusion, male and female boreal toads matured at 2 and 4 yr of age, respectively, and heavier females were more likely to produce eggs. To enhance breeding success, males should be hibernated and treated with LHRHa. In contrast, female productivity was enhanced by improving their body condition instead of subjecting them to hibernation prior to LHRHa treatment.     

Acanthocephalus bufonis (Acanthocephala) from Bufo marinus (Bufonidae: Amphibia) in Hawaii.

During a survey of the helminth parasites of the introduced toad, Bufo marinus, on O'ahu, Hawaii, an acanthocephalan corresponding to Acanthocephalus bufonis (Shipley, 1903) was found in the intestinal tract. This is a new host and locality record for A. bufonis which has only previously been recorded from amphibians in the Orient. Possible mechanisms for the introduction of A. bufonis to Hawaii, and its transmission to the toad, are discussed. Almost 98% of toads were infected with a mean intensity of 28.6 acanthocephalans per infected toad. There was a significant negative correlation between host length and intensity of infection with subadult toads having significantly higher infection levels than adult male and female toads. Trunk length of both male and female acanthocephalans was significantly related to host length.     

Seasonal changes in the migratory behaviour of the toad Bufo bufo: direction and magnitude of movements

Summary The migratory behaviour of the toad Bufo bufo was studied from February 1985 to April 1986 in the submontane region of Bavaria, West Germany. Toads were fitted with a mechanical tracking device to record individual paths of migration. Three aspects of migratory behaviour were quantified: orientation in relation to the breeding site, straightness of path, and locomotory activity. The annual activity period began with migration from the hibernation sites to the breeding pond in April. The paths went straight towards the breeding pond independent of the distance (70–420 m). During the period of oviposition the preference for the breeding site direction vanished and toads moved away from the breeding pond, but in less straight paths than before. In summer migratory activity decreased considerably and was restricted to small areas, the home ranges, at distances of 55–1600 m from the natal breeding pond. The straightness of path was rather low, because toads often returned to their starting points. During rainy nights toads occasionally left their home ranges for extensive excursions. In autumn most toads again migrated towards the breeding pond, but paths were significantly less straight and direct than in spring. However, toads stopped before reaching the breeding pond and hibernated in holes or under the leaf layer. The mortality rate of tracked toads was about 45%. The relative influence of 17 environmental variables on locomotory activity was evaluated by principal component analysis and stepwise multiple regression. Temperature at night and rainfall variables accounted for significant amounts of variance, whereas temperature by day, air humidity, and atmospheric pressure showed no correlation. Activity decreased if temperature approached 0° C or after long periods without rainfall. Within a certain range of tolerance, however, the locomotory activity of the toads was widely independent of environmental factors, indicating that endogeneous factors are more important sources of variation in the migratory behaviour of these toads than commonly assumed.     

Sex and age differences in habitat use by invasive cane toads (Rhinella marina) and a native anuran (Cyclorana australis) in the Australian wet–dry tropics

Although generalized habitat use may contribute to the success of invasive taxa, even species that are typically described as habitat generalists exhibit non‐random patterns of habitat use. We measured abiotic and biotic factors in 42 plots (each 100 × 10 m) along a 4.2‐km long unpaved road in tropical Australia, at a site that had been invaded by cane toads (Rhinella marina Bufonidae) seven years previously. We also counted anurans at night in each of these plots on 103 nights during the tropical wet season, over a five‐year period, beginning soon after the initial toad invasion. Spatial distributions differed significantly among adult male toads (n = 1047), adult female toads (n = 1222), juvenile toads (n = 342) and native frogs (Cyclorana australis Hylidae, n = 234). Adult male toads were closely associated with water bodies used as calling and/or spawning sites, whereas adult female toads and native frogs were most commonly encountered in drier forested areas on sloping ground. Juvenile toads used the margins of the floodplain more than conspecific adults did, but the floodplain itself was rarely used. Understanding which components of the habitat are most important to specific age and sex classes within a population, or how invasive species differ from native species in this respect, can clarify issues such as the spatial and temporal location of ecological impact by an invader, and the most effective places for control of the invader with minimal collateral effects on the native biota.     

Behavioural thermoregulation of the Andean toad (Bufo spinulosus) at high altitudes

Summary The body temperature of free-ranging Andean toadsBufo spinulosus was measured either directly or radiotelemetrically during two 15-day periods at 3200 m elevation in the Mantaro Valley, Central Perú. All toads attempted to maintain their diurnal sum of body temperature within a narrow range. Consequently thermoregulatory behaviour differed according to cloud cover and precipitation. If the sky was clear, toads emerged from their hiding place and exposed themselves to solar radiation during 3–5 h in the morning. Core temperature increased up to 15° C above the air temperature in shade and reached maximum values of about 32° C. At air temperatures (in sun) exceeding 29° C, toads maintained body temperatures below 32° C by evaporative cooling. Following heliothermic heating during the moring toads retreated to the shade, thereby decreasing body temperature below air temperature. Under overcast sky toads remained exposed during the whole day displaying body temperatures at or slightly above ambient levels. Quantitative models to predict the core temperature of toads under the different weather conditions demonstrated that the substrate temperature was the main energy source accounting for 64.6–77.9% of total variance whereas air temperature was of minor importance (1.5–4.4%). The unexplained variance was probably due to evaporative cooling. The volume of urine stored into the urinary bladder of toads varied diurnally; during basking in the morning hours most bladders contained large volumes of urine, whereas during the afternoon the bladders were mostly empty. The bladder contents probably serve as water reserves during basking when evaporative water loss was high. Toads preferred sites that provided shady hiding places as well as sun-exposed bare soil within a radius of 5 m. However, they frequently changed their centers of activity and moved to other sites in 20–70 m distance after periods of 2–5 days. The helio-and thigmothermic behaviour of the Andean toad permits the maintenance of high core temperature during morning which probably increases the digestion rate and accelerate growth. Evaporative cooling and preference of shady sites were employed to regulate body temperature below the morning levels in response to the constraints of water balance. Periodic changes between thigmothermic behaviour and locomotory activity during the night maintains body temperature above air temperature and prolongs the period of food uptake.Dedicated to Prof. Dr. H. Schneider on the occasion of his sixtieth birthday     

Mapping the Relative Probability of Common Toad Occurrence in Terrestrial Lowland Farm Habitat in the United Kingdom

Introduction

The common toad (Bufo bufo) is of increasing conservation concern in the United Kingdom (UK) due to dramatic population declines occurring in the past century. Many of these population declines coincided with reductions in both terrestrial and aquatic habitat availability and quality and have been primarily attributed to the effect of agricultural land conversion (of natural and semi-natural habitats to arable and pasture fields) and pond drainage. However, there is little evidence available to link habitat availability with common toad population declines, especially when examined at a broad landscape scale. Assessing such patterns of population declines at the landscape scale, for instance, require an understanding of how this species uses terrestrial habitat.

Methods

We intensively studied the terrestrial resource selection of a large population of common toads in Oxfordshire, England, UK. Adult common toads were fitted with passive integrated transponder (PIT) tags to allow detection in the terrestrial environment using a portable PIT antenna once toads left the pond and before going into hibernation (April/May-October 2012 and 2013). We developed a population-level resource selection function (RSF) to assess the relative probability of toad occurrence in the terrestrial environment by collecting location data for 90 recaptured toads.

Results

The predicted relative probability of toad occurrence for this population was greatest in wooded habitat near to water bodies; relative probability of occurrence declined dramatically > 50 m from these habitats. Toads also tended to select habitat near to their breeding pond and toad occurrence was negatively related to urban environments.     

Effect of thyroxine on some digestive enzymes of the adult male toad,Bufo melanostictus

Thyroxine is known to play an important role during the developmental process of amphibians. The present work is designed on the hypothesis that a functional relationship exists between the thyroid gland and digestive physiology in the adult toad. Three doses of thyroxine (25 microg/100 g; 50 microg/100 g; 100 microg/100 microg body weight) were orally administered daily to the adult male toad, Bufo melanostictus and changes in -amylase, pepsin, trypsin, and esterase activities of the different zones of the digestive tract, pancreas and liver were observed. The observations were made on 7, 15, and 30 days of thyroxine treatment. The result shows that the dose of 25 microg increases the enzyme activities in the gastrointestinal tract, liver and pancreas of 7 days treatment. However, prolonged treatment with all the doses shows little effect on these enzymes. The pattern of changes in the enzymatic activities in the digestive tract of the adult male toads show more advanced compartmentalization than that of fishes. It is also found that site-specific enzyme production is not pronounced in this amphibious vertebrate. It is proposed that a positive functional relationship between thyroxine and digestive enzymes exists in the adult male toad.     

Effects of the ecological conditions on morphological variations of the green toad, Bufo viridis, in Turkey

Ten populations of the green toad, Bufo viridis, from three areas having different climatic regimes were compared. Multivariate analyses of morphological characteristics revealed three distinct populations of green toads: the first group inhabiting the lowland and humid localities was characterized by a small body size, the second group occurred at higher altitudes and in desert areas had a significantly larger body size, while the third group from the Mediterranean localities was characterized by a large tympanum, a longer tarsus and short first toes. Interestingly, the specimens from Giresun Island were clustered with the second group with a similar pattern of body size. Differences in the morphological characteristics among populations were positively associated with ecological conditions, and not with geographic distance. Some characteristics previously used for taxonomic purposes in B. viridis were shown to be associated with local climatic conditions.     

Allometry and selection in a novel predator–prey system: Australian snakes and the invading cane toad

Because many organismal traits vary with body size, interactions between species can be affected by the respective body sizes of the participants. We focus on a novel predator–prey system involving an introduced, highly toxic anuran (the cane toad, Bufo marinus ) and native Australian snakes. The chance of a snake dying after ingesting a toad depends on the size of the snake and the size of the toad, and ultimately reflects the effect of four allometries: (1) physiological tolerance (the rate that physiological tolerance to toad toxin changes with snake size); (2) swallowing ability (the rate that maximal ingestible toad size (i.e. snake head size) increases with snake body size); (3) prey size (the rate that prey size taken by snakes increases with snake head size) and (4) toad toxicity (the rate that toxicity increases with toad size). We measured these allometries, and combined them to estimate the rate at which a snake's resistance changes with toad toxicity. The parotoid glands (and thus, toxicity) of toads increased disproportionately with toad size (i.e. relative to body size, larger toads were more toxic) but simultaneously, head size relative to body size (and thus, maximal ingestible prey size relative to predator size) declined with increasing body size in snakes. Thus, these two allometries tended to cancel each other out. Physiological tolerance to toxins did not vary with snake body size. The end result was that across snake species, mean adult body size did not affect vulnerability. Within species, however, smaller predators were more vulnerable, because the intraspecific rate of decrease in relative head size of snakes was steeper than the rate of increase in toxicity of toads. Thus, toad invasion may cause disproportionate mortality of juvenile snakes, and adults of the sex with smaller mean adult body sizes.     

The Influence of Parasite Infection on Mating Success in Spadefoot Toads, Scaphiopus couchii

The desert toads, Scaphiopus couchii, have an annual activityseason of less than 8 weeks and experience only one significantparasite infection: the monogenean Pseudodiplorchis americanusis transmitted during host spawning and provides a natural systemfor testing the influence of parasite burden on host matingsuccess. The 10 month hibernation involves total starvationduring which the blood-feeding parasites reduce fat reservesand haematocrit. The toads emerge and spawn on the first nightafter rainfall, before they replenish depleted reserves. Malechorusing is energetically very demanding and mate selection,limited to a 7 hr nocturnal assembly, is determined principallyby female choice. Parasite transmission triggered by host sexualactivity results in 100% prevalence and high intensities ofinfection amongst males. Around 50% of toads lose their burdensentirely but the rest carry chronic infections throughout hosthibernation. Field data show a consistent reduction in intensityeach year strongly suggesting resistance. Parasite infectionis pathogenic and creates extra stress during hibernation; therefore,to the extent that elimination of infection is heritable, toadsentering spawning assemblies with heavy burdens should makepoor mates. However, extensive field studies show no correlationbetween mate success and parasite burden. Although infectioncan prejudice survival, it is only one of several inter-relatedfactors (including feeding success, tolerance of hibernation).The condition of successful and unsuccessful males in spawningassemblies indicates that all exceed a threshold at which parasite-inducedpathology is significant. Males which are debilitated by infection—orother factors—are selected against before mate choicebegins