Comparisons of blood viscosity between amphibians and mammals at 3°C and 38°C

(1) The range of temperature exposure of endotherms is narrow compared to ectotherms that can experience daily and seasonal temperature fluxes. (2) Comparison of the blood viscosity of amphibians (bullfrog, Woodhouse's toad, and marine toad) and mammals (horse, dog, and rat) at 3°C and 38°C was undertaken to determine if the effect of temperature on blood viscosity was diminished in amphibians relative to mammals. (3) Mammals did not consistently show greater changes in blood viscosity, plasma viscosity, or relative viscosity with decreasing temperatures relative to the amphibians in this study. (4) These data do not support our hypothesis that blood viscosity of amphibians is less affected by temperature than mammalian blood.     

Two serine protease inhibitors from the skin secretions of the toad, Bombina microdeladigitora

Two serine protease inhibitors (named BMSI 1 and BMSI 2, respectively) were identified from the skin secretions of the toad, Bombina microdeladigitora. The cDNAs encoding BMSIs were cloned from a cDNA library prepared from the toad skin. The deduced complete amino acid sequences of BMSIs indicate that mature BMSI 1 and BMSI 2 are composed of 60 amino acids including 10 half-cystines to form 5 disulfide bridges. A FASTA search in the databanks revealed that BMSIs exhibit sequence similarity with other serine protease inhibitors from amphibians of the genus Bombina. BMSI 1 potently inhibited trypsin and thrombin with a K(i) value of 0.02 μM and 0.15 μM, respectively. Sequence analysis revealed that all serine protease inhibitors from five amphibians of the genus Bombina share highly conserved primary structures.     

长白山国家级自然保护区两栖类动物道路致死特征分析

为了解旅游道路交通导致的两栖类致死效应,于2007~2014年在环绕长白山国家级自然保护区的环长白山旅游公路(环区公路)、头道环区公路以及白山公路分别选取1条样线,采用直接计数法统计了交通致死的两栖类种类、数量,以及致死个体处于交通路段的位置,共统计257次,累计调查样带总长2 441 km。道路致死两栖类种类有5种,累计致死个体数为7 910只,平均致死率为3.34只/km。最容易致死的两栖类动物是中国林蛙(Rana chensinensi)、东方铃蟾(Bombina orientalis)、中华蟾蜍(Bufo gargariza),这3种动物占到致死总数的96.36%;经Moses Test检验,白山公路的致死率、头道公路的致死率及环区公路致死率均与对照土路致死率差异显著(白山公路:n=67,P=0.000;头道公路:n=76,P=0.000;环区公路:n=164,P=0.000)。白山公路与头道公路两栖类致死率差异显著(n=93,P=0.010),白山公路与环区公路的两栖类道路致死率差异显著(n=181,P=0.000);头道公路与环区公路的两栖类道路致死率差异显著(n=190,P=0.000)。两栖类道路致死高峰期在8月份,其次较集中出现在4~6月的繁殖期。从公路年度致死变化来看,2007~2014年期间除了中国林蛙和中华蟾蜍致死率呈小幅波动外,其他种类致死率总体呈下降趋势,只有日本树蟾(Hyla japonica)年度致死率差异显著(Kruskal Wallis Test:χ2=18.031,df=7,P=0.012);不同月份中华蟾蜍(Pearson:R=0.939,n=7,P=0.002)、中国林蛙(Pearson:R=0.912,n=7,P=0.004)、日本树蟾(Pearson:R=0.904,n=7,P=0.005)致死率与交通量显著相关。本研究认为,两栖类的道路致死率与两栖类种群大小、种群密度、生活史及交通量等都存在显著相关性。     

Seasonal exploitation of amphibians by otters (Lutra lutra) in north-east Scotland

The dietary importance of amphibians to otters was studied in north-east Scotland in 1987. The proportion of amphibians identified in otter droppings varies seasonally and according to the habitat. The consumption of amphibians increases in late winter and spring while they are either hibernating or spawning. Estimations of amphibian population density at spawning sites have been carried out. There is a significant correlation between the availability of amphibians and their consumption by otters. The main amphibian species involved in the diet is the common frog, Rana temporaria (60.8%), while the common toad, Bufo bufo , represents only 1.7% of the amphibian prey.     

A neurophysiological analysis of the forms of wakefulness and rest in the toad Bufo bufo and the frog Rana ridibunda in the circadian cycle

Studies have been made on the spectral EEG parameters in the forebrain and midbrain of the toad and frog during active and passive forms of natural behaviour within a diurnal cycle. Basing on evaluation of animal reactivity and on EEG analysis performed by presentation of the power of EEG signals via the frequency spectrum, it is possible to obtain a reliable identification of functional condition of amphibians. Behavioural and EEG patterns of 3 forms of wakefulness and 2 forms of rest are presented for frogs and toads.     

Highlighting the effects of land-use change on a threatened amphibian in a human-dominated landscape

In Western Europe, habitat loss and landscape fragmentation has led to significant population decline in various animal groups, including amphibians. The extinction of the last natural populations of the yellow-bellied toad in Belgium, Luxembourg and several regions of southern and western France suggests a widespread decline. By using site-occupancy models and adding covariates corresponding to the human-influenced features of the landscape, we tried to identify the relative effects of different land-use types on the species’ distribution pattern in a man-made environment (the Alsatian Rhine floodplain in France). We recorded presence–absence data in 150 forest sample plots (300 × 300 m) and then modeled species distribution while taking into account detection errors in the field. Land-use was recorded on two spatial scales: within the forest sample plots and in a 1500 m radius buffer area around the forest plots. In the forest plots, toad occurrence was negatively correlated with loss of forest cover to agricultural land. In contrast, occurrence is positively correlated with the density of human-made rutted dirt paths and tracks, which provide semi-natural breeding sites. In the 1500 m radius buffer zones around forest plots, toad occurrence was negatively correlated with the density of urbanization and road networks. These results can be used to plan conservation strategies for amphibians in human-dominated landscapes.     

Population differentiation of temperate amphibians in unpredictable environments

Amphibians are a globally distributed and diverse lineage, but much of our current understanding of their population genetic structure comes from studies in mesic temperate habitats. We characterize the population genetic structure of two sympatric explosive breeding amphibians in the southwestern deserts of the United States: the Great Plains toad ( Anaxyrus cognatus ) and Couch's spadefoot toad ( Scaphiopus couchii ). For both species, we find limited genetic differentiation even between populations in adjacent valleys separated by dispersal barriers such as mountainous habitats. To understand how population genetic patterns in these two arid-adapted species compare to taxa in more mesic environments, we computed a standardized measure of population differentiation for A. cognatus , S. couchii , and for pond-breeding amphibians that inhabit mesic temperate environments. Our results indicate that the arid-adapted species have lower population genetic structure at fine and moderate scales than most other amphibian species we surveyed. We hypothesize that stochasticity in the availability of appropriate breeding sites as well as landscape homogeneity may result in increased population connectivity in desert-adapted frogs. Future work examining fine-scale population structure in amphibians from a diversity of habitats will test the generality of our findings. Intraspecific comparisons among localities with varied seasonality and habitats will be particularly useful for investigating the interaction between species-typical population dynamics and environmental characteristics as determinants of population connectivity in pond-breeding amphibians.     

基于人工掩蔽物法的若尔盖湿地中华蟾蜍种群生态研究

近几十年来,全球两栖动物种群衰减显著,两栖动物的生存现状引起了越来越多的生态学家和保护生物学家的关注。若尔盖湿地不仅是世界上最大的一块高原泥炭沼泽湿地,也是我国生物多样性保护的热点地区之一。该地区分布有3种两栖类:高原林蛙(Rana kukunoris)、倭蛙(Narorana pleskei)和中华蟾蜍岷山亚种(Bufo gargarizans minshanica)。已有研究发现该3种两栖动物种群数量均有不同程度的下降。采用人工掩蔽物法,在2011—2014年对该地区中华蟾蜍种群生态做了连续追踪。结果表明:该区域中华蟾蜍种群数量逐年波动较大,年龄结构数据显示该种群处于增长期;中华蟾蜍为聚集分布,且发现率与水体距离呈显著的线形关系(P0.01),在样地范围内离水体越远,发现的个体越多;线形样方比方形样方捕获动物的效率更高(P=0.018);中华蟾蜍亚成体的肥满度季节间无明显差异;中华蟾蜍具有较强的迁徙能力,可能沿着固定的线路周而复始的迁徙。对于家域范围大,迁徙距离远的中华蟾蜍这类物种,应加强最适栖息地的保护并防止栖息地破碎化。     

Myiasis by Lucilia silvarum (Calliphoridae) in amphibian species in boreal Alberta, Canada

We report myiasis by Lucilia silvarum with an overall prevalence of 0.9% in amphibian populations in boreal Alberta. In the period 1998--1999, we documented L. silvarum infestations in wild populations of wood frog (Rana sylvatica), boreal chorus frog (Pseudacris maculata), boreal toad (Bufo boreas boreas), and Canadian toad (B. hemiophrys). We believe this is the first record of this parasite from boreal and Canadian toads. Almost all previous records of L. silvarum parasitism in North America indicate that myiasis is fatal to an anuran host. Here, we provide the first record of adult individuals from 2 species (wood frog and boreal toad) surviving infestations. Although we actively captured and examined amphibians in Alberta from 1996 to 2006, we only found parasitism in 1998 and 1999. This is the most northerly record of anuran infestations by this parasite.     

Amphibians as definitive hosts for pentastomids: Raillietiella bufonis n.sp. from Bufo lemur in Puerto Rico and a reassessment of Raillietiella indica Gedoelst, 1921

Summary A new species of a cephalobaenid pentastomid infecting the lungs of a toad, Bufo lemur, from Puerto Rico is described. The single infection comprises all stages from infective larvae to mature males and females and therefore we consider the toad to be a definitive host. We reassess the evidence concerning Raillietiella indica Gedoelst, 1921, another raillietiellid from the lungs of an Indian toad, B. melanostictus which, hitherto, has been considered by most authors an immature stage from an intermediate host. It now appears that the type specimen was probably a gravid female and that R. indica is a valid species which also matures in a toad. Thus amphibians are established as a new class of definitive hosts for pentastomids. ac]19820218     

Multi-event models reveal the absence of interaction between an invasive frog and a native endangered amphibian

Among the multiple factors involved in the decline of amphibians, competition with alien species remains understudied. In Western Europe, non-native marsh frogs (Pelophylax ridibundus and other sister species) have been widely imported for gastronomic purposes and have then often escaped from captivity. Apart from closely related species, the impact of the subsequent colonization of non-native species on native amphibians is still unknown. In this study, we analysed the response of a threatened species, the yellow-bellied toad (Bombina variegata), faced with the presence of alien frogs in mountain rivers in France. We studied the co-occurrence pattern of the native toad and the alien frog, while taking into account co-dependence of the detection rates in both species. We tested three main scenarios to explain a non-random pond occurrence for each species and to establish their respective habitat preferenda: (1) a pond-area-dependent scenario, predicting different responses from each species for a given pond size, (2) a fish-dependent scenario, predicting different responses from each species given the presence of fish in ponds, and (3) a floodplain-width-dependent scenario, predicting different responses from each species given the geomorphological characteristics of the floodplain. Taking into account site-specific covariates, we concluded that introduced frogs do not currently have an impact on the native population of the yellow-bellied toad.     

Histochemistry of the juxtaglomerular apparatus in the toad Bufo bufo

Summary An investigation regarding the question of whether there exists a macula densa as part of the juxtaglomerular apparatus in the kidney of amphibians has been carried out. With the aid of a histochemical reaction for glucose-6-phosphate dehydrogenase activity, the presence of a macula densa zone as a specialized part of the distal tubule in the toad Bufo bufo was demonstrated. The functional significance of the high glucose-6-phosphate dehydrogenase activity in the macula densa cells is discussed.     

Calbindin D28k is essentially located in the colonic part of the toad intestine

The distribution of calbindin D28k in the digestive system and the urinary bladder of the toad was investigated using immunohistochemistry and Western blotting. By analogy with mammals and birds, the protein was expected to be located preferentially in the duodenal part of the intestine. Interestingly, absorptive cells of the duodenum were totally devoid of calbindin D28k while the colon contained high amounts of the calcium-binding protein. This reversed polarity of calbindin D28k content in the toad intestine should obviously correspond to a different scheme of calcium absorption regulation between amphibians and higher vertebrates. Calbindin D28k containing neuroendocrine-like cells were found scattered in the proximal parts of the gut with a similar distribution to what has been described in rat and chick intestine. The oesophagus, the stomach, and the intrinsic nervous sytem of the intestine were negative. No significant amounts of the proteins were found in the urinary bladder, which is known to be a site of Ca²⁺ active transport.     

Mortality of vertebrates on a road crossing the Biebrza Valley (NE Poland)

Mortality of vertebrates was monitored on a local road running across Poland’s Biebrza River Valley during 2 years (August 2005–July 2006). On the basis of distance from the river and surrounding habitats, the road (of total length 2,510 m) was divided into three stretches. The road was monitored on foot by two people every month, over a few consecutive days. A total of 1,892 road kills representing at least 47 species were found. Of these, 90.7% were amphibians, 4.2% mammals, 3.1% birds and 2.0% reptiles. Most (70%) of the amphibians were anurans, with the common toad, common frog and moor frog among them together accounting for 82% of the total. Mortality among amphibians differed between months, most anurans dying in May and August, while a majority of Urodela are lost in October. The peaks in mortality were connected with the migration of adult amphibians in spring and juveniles in summer and autumn. The number of amphibians killed was greatest on the (wettest) stretch adjacent to the river and decreased with distance from it. Mortality among birds was highest in July—probably in association with the dispersal of young individuals. Among recorded mammalian road kills, there was a prevalence of small rodents (mainly voles) and insectivores (mainly shrews). Medium-sized mammals were found only accidentally. Mortality in general was conditioned by the number of anurans killed.     

Diversity of opsin immunoreactivities in the extraretinal tissues of four anuran amphibians

The pineal complex, deep brain, and skin have been known to function as extraretinal photoreceptors in non-mammalian vertebrates. To see the diversity of localization of extraretinal photoreceptors in lower vertebrates having different habitats, we analyzed the opsin-like immunoreactivities in anuran amphibians, Xenopus laevis, Rana catesbeiana, Rana nigromaculata, and Bufo japonicus. An antiserum (toad Rh-AS) was raised against rhodopsin purified from the retinas of Japanese toad, B. japonicus. In the retina of all the anurans examined, the outer segments of rods were immunopositive to toad Rh-AS. The outer segments of most pinealocytes were immunopositive in R. catesbeiana, R. nigromaculata, and B. japonicus. The outer segments of photoreceptor-like cells within the frontal organ of R. nigromaculata were immunostained. Interestingly, toad Rh-AS immunostained many secretory cells of mucous glands in the head skin of B. japonicus, implying the presence of a novel photoreceptive molecule. Within the hypothalamus, toad Rh-AS immunostained many cells in the magnocellular preoptic nucleus of R. catesbeiana and B. japonicus. Toad Rh-AS also labeled cerebrospinal fluid (CSF)-contacting cells in the anterior preoptic nucleus of R. nigromaculata and those adjacent to the lateral ventricle within the septum of R. catesbeiana. Thus the distribution patterns of the rhodopsin-like immunoreactivities among the anurans were highly diverged, and there was no relationship between the distribution patterns and their habitats. J. Exp. Zool. 286:136-142, 2000.     

A suspected parasite spill-back of two novel Myxidium spp. (Myxosporea) causing disease in Australian endemic frogs found in the invasive Cane toad

Infectious diseases are contributing to the decline of endangered amphibians. We identified myxosporean parasites, Myxidium spp. (Myxosporea: Myxozoa), in the brain and liver of declining native frogs, the Green and Golden Bell frog (Litoria aurea) and the Southern Bell frog (Litoria raniformis). We unequivocally identified two Myxidium spp. (both generalist) affecting Australian native frogs and the invasive Cane toad (Bufo marinus, syn. Rhinella marina) and demonstrated their association with disease. Our study tested the identity of Myxidium spp. within native frogs and the invasive Cane toad (brought to Australia in 1935, via Hawaii) to resolve the question whether the Cane toad introduced them to Australia. We showed that the Australian brain and liver Myxidium spp. differed 9%, 7%, 34% and 37% at the small subunit rDNA, large subunit rDNA, internal transcribed spacers 1 and 2, but were distinct from Myxidium cf. immersum from Cane toads in Brazil. Plotting minimum within-group distance against maximum intra-group distance confirmed their independent evolutionary trajectory. Transmission electron microscopy revealed that the brain stages localize inside axons. Myxospores were morphologically indistinguishable, therefore genetic characterisation was necessary to recognise these cryptic species. It is unlikely that the Cane toad brought the myxosporean parasites to Australia, because the parasites were not found in 261 Hawaiian Cane toads. Instead, these data support the enemy-release hypothesis predicting that not all parasites are translocated with their hosts and suggest that the Cane toad may have played an important spill-back role in their emergence and facilitated their dissemination. This work emphasizes the importance of accurate species identification of pathogens relevant to wildlife management and disease control. In our case it is paving the road for the spill-back role of the Cane toad and the parasite emergence.     

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.     

Tachykinin peptides and receptors: putting amphibians into perspective

The tachykinins form one of the largest peptide families in nature. In this review, we describe the comparative features of the tachykinin peptides and their receptors, focusing particularly on amphibians. We also summarize our systematic studies of the localization, characteristics, and actions of bufokinin, a toad substance P-related peptide, in its species of origin. In addition, we discuss the establishment of multiple isoforms of the NK1-like receptor in the toad, and their structure, pharmacology and tissue distributions. We conclude that tachykinin peptides and receptors are well conserved in terms of their structures, physiological functions and coupling mechanisms during tetrapod evolution.     

The smooth muscle pharmacology of maximakinin, a receptor-selective, bradykinin-related nonadecapeptide from the venom of the Chinese toad, Bombina maxima

Structural homologues of vertebrate regulatory peptides found in defensive skin secretions of anuran amphibians often display enhanced bioactivity and receptor binding when compared with endogenous mammalian peptide ligands. Maximakinin, a novel N-terminally extended bradykinin (DLPKINRKGPRPPGFSPFR) from the skin venom of a Chinese toad (Bombina maxima), displays such activity enhancement when compared with bradykinin but is additionally highly selective for mammalian arterial smooth muscle bradykinin receptors displaying a 50-fold increase in molar potency in this smooth muscle type. In contrast, a 100-fold decrease in molar potency was observed at bradykinin receptors in intestinal and uterine smooth muscle preparations. Maximakinin has thus evolved as a "smart" defensive weapon in the toad with receptor/tissue selective targeting. Natural selection of amphibian skin venom peptides for antipredator defence, through inter-species delivery by an exogenous secretory mode, produces subtle structural stabilisation modifications that can potentially provide new insights for the design of selectively targeted peptide therapeutics.     

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.