Observations on myiasis by the calliphorids, Bufolucilia silvarum and Bufolucilia elongata, in wood frogs, Rana sylvatica, from southeastern Wisconsin

Larvae of certain species of blowflies (Calliphoridae) can cause myiasis in frogs and toads, but there are few reports from North American amphibians. Of these, most are from toads (bufonids). In this study, we observe primary myiasis in a population of juvenile wood frogs, Rana sylvatica, collected on 22-23 August 2003, from southeastern Wisconsin and compare our observations with previous studies on myiasis from toads. Two (5%) of 39 frogs were infected by the blow fly Bufolucilia silvarum, with an intensity of 28 and 31, whereas 1 (2.5%) of 39 frogs was infected by the blow fly Bufolucilia elongata with an intensity of 14. We found that (1) B. silvarum lay eggs on healthy wood frogs, (2) eggs hatch, with first-instar maggots penetrating under the skin, (3) maggots develop to mature third instars within 13-16 hr of egg hatching, (4) maggots kill the host within 7-47 hr of egg hatching, and (5) maggots consume the entire frog carcass reducing it to bones within 42-59 hr of egg hatching. Our observations on the time of death and how quickly carcasses of wood frogs were consumed by these maggots compared with previous studies on toads suggest that finding infected juvenile wood frogs may be uncommon. Therefore, myiasis by these flies on wood frogs and other small terrestrial anurans may be a phenomenon that is much more common than is currently observed. This is the first report of B. silvarum and B. elongata causing myiasis in wood frogs.     

Observations on myiasis by the calliphorid,Bufolucilia silvarum,in the eastern American toad (Bufo americanus americanus) from southeastern Wisconsin

Larvae of certain species of blowflies (Calliphoridae) cause myiasis in amphibians which may result in significant mortality, yet there are few reports from North America. In this study, we observed primary myiasis in a population of juvenile eastern American toads (Bufo americanus americanus) collected during May-July 1998 from southeastern Wisconsin (USA). Nine (6%) of 140 toads were infected by the green blow fly (Bufolucilia silvarum) with a mean intensity of 10.5 +/- 7.2 (range = 1-24). Weekly parasite prevalence and mean intensity remained low, ranging from 0-20% and 2 +/- 1.4 to 14 +/- 6, respectively. We found: 1) flies lay eggs on healthy toads, 2) eggs hatch with first instar maggots penetrating under the skin, 3) maggots develop to mature third instars within 5-7 days, 4) maggots leave the host and form pupa within 8-11 days of hatching, and 5) maggots pupate within 7-9 days at room temperature. All infected toads died within 1-2 wk as a result of the infection. The low prevalence observed in this study and other reports of this species from mammalian and bird carcasses indicated that B. silvarum is probably a facultative parasite of toads and other amphibians in the United States. This is the first report of B. silvarum causing myiasis in Wisconsin amphibians and the first report in eastern American toads in the United States.     

Stability and permanence in gender- and stage-structured models for the boreal toad

The boreal toad Bufo boreas boreas, once common in the western USA, is listed as an endangered species in Colorado and New Mexico, and protected in Wyoming. Populations have dramatically declined due to the presence of the fungal pathogen Batrachochytrium dendrobatidis (Bd). A gender- and stage-structured model for the boreal toad is formulated which depends on its life cycle and breeding strategies. In addition, an epizootic model for the spread of Bd is formulated. Analysis of these models provides two thresholds. The first threshold, the basic reproduction number for the population, ?(0), determines whether the population persists and the second threshold, the basic reproduction number for the fungal disease, ?(F), determines whether the disease persists. If ?(0)>1 and ?(F)<1, then the population becomes disease-free. However, if both thresholds are greater than one, the population levels are severely reduced by the fungal pathogen.     

Survival of three species of anuran metamorphs exposed to UV-B radiation and the pathogenic fungus Batrachochytrium dendrobatidis

When exploring the possible factors contributing to population declines, it is necessary to consider multiple, interacting environmental stressors. Here, we investigate the impact of 2 factors, ultraviolet radiation and disease, on the survival of anuran amphibians. Exposure to ultraviolet-B (UV-B) radiation increases mortality and results in various sub-lethal effects for many amphibian species. Infectious diseases can also negatively impact amphibian populations. In this study, we exposed metamorphic individuals (metamorphs) to both UV-B and Batrachochytrium dendrobatidis (BD), a fungal pathogen and cause of the disease chytridiomycosis, and monitored survival for 3 wk. We tested for possible interactions between UV-B and BD in 3 species: the Cascades frog Rana cascadae; the Western toad Bufo boreas; and the Pacific treefrog Hyla regilla. We found strong interspecific differences in susceptibility to BD. For example, R. cascadae suffered a large increase in mortality when exposed to BD; B. boreas also experienced mortality, but this effect was small relative to the R. cascadae response. H. regilla did not show any decrease in survival when exposed to either factor. No synergistic interactions between UV-B and BD were found for any of the test species. A previous study investigating the impact of BD on larval amphibians showed different species responses (Blaustein et al. 2005a). Our results highlight the importance of studying multiple life history stages when determining the impact of environmental stressors. The contrast between these 2 studies emphasizes how vulnerability to a pathogen can vary between life history stages within a single species.     

Effects of the landscape on boreal toad gene flow: does the pattern–process relationship hold true across distinct landscapes at the northern range margin?

Understanding the impact of natural and anthropogenic landscape features on population connectivity is a major goal in evolutionary ecology and conservation. Discovery of dispersal barriers is important for predicting population responses to landscape and environmental changes, particularly for populations at geographic range margins. We used a landscape genetics approach to quantify the effects of landscape features on gene flow and connectivity of boreal toad (Bufo boreas) populations from two distinct landscapes in south-east Alaska (Admiralty Island, ANM, and the Chilkat River Valley, CRV). We used two common methodologies for calculating resistance distances in landscape genetics studies (resistance based on least-cost paths and circuit theory). We found a strong effect of saltwater on genetic distance of CRV populations, but no landscape effects were found for the ANM populations. Our discordant results show the importance of examining multiple landscapes that differ in the variability of their features, to maximize detectability of underlying processes and allow results to be broadly applicable across regions. Saltwater serves as a physiological barrier to boreal toad gene flow and affects populations on a small geographic scale, yet there appear to be few other barriers to toad dispersal in this intact northern region.     

Variation in anti-parasite behaviour and infection among larval amphibian species

Along with immune defences, many animals exhibit effective anti-parasite behaviours such as parasite avoidance and removal that influence their susceptibility to infection. Host ecology and life history influence investment into comparatively fixed defences such as innate immunity but may affect the strength of anti-parasite behaviours as well. We investigated activity levels in five different species of larval amphibian with varying life histories and ecology in control, novel food stimulus, and trematode parasite (Echinoparyphium sp.) threat conditions. There was a significant interaction of species and treatment given that American toad (Bufo americanus), wood frog (Lithobates sylvaticus), and bullfrog (Lithobates catesbeianus) tadpoles generally increased their activity when parasite infectious stages were present while grey tree frogs (Hyla versicolor) and northern leopard frogs (Lithobates pipiens) did not, even though activity was negatively related to infection. In addition, there was considerable variation among species in their susceptibility to parasitism, with infection prevalence ranging from 17 % in bullfrog tadpoles to 70 % in wood frogs. However, amphibian life history (larval and adult traits) was not related to parasitism or level of anti-parasite behaviour at the species level. Consequently, we suggest that future investigations include more species with a range of life history traits and also consider host ecology, particularly if conspicuous anti-parasite behaviours are more likely in amphibian species that experience a relatively low risk of predation.     

The role of pollutant accumulation in determining the use of stormwater ponds by amphibians

Urbanization often results in the creation of habitats such as stormwater management ponds. Although stormwater ponds are designed to retain runoff and associated pollutants, they are frequently colonized by wildlife including pond-breeding amphibians. Understanding of the ecological function of these created habitats is limited. This study investigated the role of pollutants in shaping use of stormwater ponds by amphibians. A survey of 68 stormwater ponds in Baltimore County, Maryland, and statistical modeling found wood frogs (Rana [= Lithobates] sylvatica) were more likely to breed in ponds with longer hydroperiods and Cl^? concentrations below approximately 250 mg/L. American toad (Bufo [= Anaxyrus] americanus) use of ponds was primarily influenced by hydroperiod; toads were more likely to use longer hydroperiod ponds. To confirm use was a result of toxicity and differential sensitivity among species, wood frog and American toad embryos and larvae were exposed to sediment from six stormwater ponds spanning the range of pollutant conditions documented in the field. Survival of wood frogs through metamorphosis was related to metal and salt levels of pond sediments, but survival of American toads was not. In agreement with the field study, no wood frog larvae survived to metamorphosis when Cl^? levels were above 260 mg/L. The results suggest that pollutants that accumulate in stormwater ponds, specifically road deicing salts, are acting as local filters capable of creating unique assemblages of anuran larvae in urban areas.     

Does restricted access limit management of invasive urban frogs?

Management recommendations that target urban invaders should consider environmental and socio-economic aspects peculiar to the urban landscape. Urbanization often leads to the fragmentation of the invaded landscape into subunits inaccessible to managers (restricted access) or for which detailed information is lacking. Using models to explore impact of these limitations on management success provides a useful approach to propose effective countermeasures. Here we deploy a spatially explicit age-structured model applied to a pond network to investigate how restricted access and lack of detailed information may affect management of three invasive anuran species across a peri-urban landscape. The target species, the guttural toad Sclerophrys gutturalis, the African clawed frog Xenopus laevis and the painted reed frog Hyperolius marmoratus, belong to different ecotypes (terrestrial, aquatic and arboreal, respectively) and have different life history traits. We show that restricted property access significantly constrains management success in two of the three species (the guttural toad and the painted reed frog), while lack of detailed information around the invaded landscape impedes successful management in only one species (the guttural toad). The species-dependent response we detected is due to contrasting demographic and spatial invasion dynamics linked to the different anuran ecotypes. Our work highlights the necessity to adopt a context-dependent approach when proposing management recommendations in urban environment.     

Clinostomum attenuatum (Digenea) from the eye of Bufo marinus

Two specimens of the cane toad, Bufo marinus, were found with single worm infestations of the eye. One worm was excysted in the eye chamber, the other still encysted in the choroid layer; both worms were identified as Clinostomum attenuatum. This finding is both a new host record and a new site of development for this amphibian parasite.     

Pond tadpoles with generalized morphology: is it time to reconsider their functional roles in aquatic communities?

With rare exceptions, anuran larvae have traditionally been considered to occupy lower trophic levels in aquatic communities where they function as microphagous suspension feeders. This view is being challenged by studies showing that tadpoles with generalized morphology often function as macrophagous predators. Here, we review the literature concerning macrophagy by tadpoles and provide two additional examples involving generalized tadpoles. In the first, we demonstrate with laboratory and field experiments that wood frog (Rana sylvatica) tadpoles are major predators of macroinvertebrates in ponds. In the second, we show that green frog (R. clamitans) tadpoles can cause catastrophic reproductive failure of the wood frog via egg predation. These results and data from other studies challenge the assumption that generalized tadpoles function as filter-feeding omnivores, and question the general applicability of community organization models which assume that predation risk increases with pond permanence. We suggest that predation risk is greater in temporary ponds than in more permanent ponds for many organisms that are vulnerable to predation by tadpoles. This being so, a conditional model based upon interactions that are species-specific, life-stage-specific, and context-dependent may better explain community organization along hydrological gradients than models which assume that temporary ponds have few or no predators. Received: 30 November 1998 / Accepted: 2 May 1999     

The toad fly Lucilia bufonivora: its evolutionary status and molecular identification

The blow fly genus Lucilia is composed largely of saprophages and facultative myasis agents, including the economically important species Lucilia cuprina (Wiedemann) (Diptera: Calliphoridae) and Lucilia sericata (Meigen). Only one species is generally recognized as an obligate agent of myiasis, Lucilia bufonivora Moniez, and this is an obligate parasite of toads. Lucilia silvarum (Meigen), a sister species, behaves mainly as a carrion breeder; however, it has also been reported as a facultative parasite of amphibians. Morphologically, these species are almost identical, and historically this has led to misidentification, taxonomic ambiguity and a paucity of studies of L. bufonivora. In this study, dipterous larvae were analysed from toad myiasis cases from the U.K., The Netherlands and Switzerland, together with adult specimens of fly species implicated in amphibian parasitism: L. bufonivora, L. silvarum and Lucilia elongata Shannon (from North America). Partial sequences of two genes, cox1 and ef1α, were amplified. Seven additional blow fly species were analysed as outgroups. Bayesian inference trees of cox1, ef1α and a combined‐gene dataset were constructed. All larvae isolated from toads were identified as L. bufonivora and no specimens of L. silvarum were implicated in amphibian myiasis. This study confirms L. silvarum and L. bufonivora as distinct sister species and provides unambiguous molecular identification of L. bufonivora.     

Grey flesh fly (Wohlfahrtia vigil) parasitism of a Preble's meadow jumping mouse (Zapus hudsonius preblei)

A Preble's meadow jumping mouse (Zapus hudsonius preblei) parasitized by five grey flesh fly (Wohlfahrtia vigil) larvae was discovered at the United States Air Force Academy in Colorado Springs, Colorado (USA), in June 1998. This is the first documented case of grey flesh fly parasitism of jumping mice (Family Dipodidae). The lesion was approximately 6 mm wide and was partially hidden under a mat of wet fur. Myiasis was found in one (0.7%) of 146 jumping mice captured at the Academy in 1998. The Preble's meadow jumping mouse is considered a threatened subspecies by the United States Fish and Wildlife Service. Although grey flesh fly myiasis can be fatal, it is unknown whether it is affecting populations of Preble's meadow jumping mouse. This is the first report of grey flesh fly myiasis in free-ranging wildlife in Colorado.     

Batrachomyia strigapes (Diptera) parasitism of Uperoleia laevigata (Anura).

To quantify the occurrence of the dipteran parasite Batrachomyia strigapes, the ground frog assemblage associated with a large ephemeral wetland in Western Sydney was sampled between January 1999 and April 2000. Parasite infection was restricted to Uperoleia laevigata (5.1%, n = 1,492). Parasites were found under the parotoid glands, most frequently on the left side. After correction for frog size, infection was shown to significantly reduce frog weight. Despite reduced individual fitness, because the level of infection was low in the population, it is unlikely to have a major effect on the anuran population under current environmental conditions.     

Long‐term monitoring of tropical alpine habitat change,Andean anurans,and chytrid fungus in the Cordillera Vilcanota,Peru: Results from a decade of study

The Cordillera Vilcanota in southern Peru is the second largest glacierized range in the tropics and home to one of the largest high‐alpine lakes, Sibinacocha (4,860 m). Here, Telmatobius marmoratus (marbled water frog), Rhinella spinulosa (Andean toad), and Pleurodema marmoratum (marbled four‐eyed frog) have expanded their range vertically within the past century to inhabit newly formed ponds created by ongoing deglaciation. These anuran populations, geographically among the highest (5,200–5,400 m) recorded globally, are being impacted by the chytrid fungus Batrachochytrium dendrobatidis (Bd), and the disease it causes, chytridiomycosis. In this study, we report results from over a decade of monitoring these three anuran species, their habitat, and Bd infection status. Our observations reveal dynamic changes in habitat including ongoing rapid deglaciation (18.4 m/year widening of a corridor between retreating glaciers from 2005 to 2015), new pond formation, changes in vegetation in amphibian habitat, and widespread occurrence of Bd in amphibians in seven sites. Three of these sites have tested positive for Bd over a 9‐ to 12‐year period. In addition, we observed a widespread reduction in T. marmoratus encounters in the Vilcanota in 2008, 2009, and 2012, while encounters increased in 2013 and 2015. Despite the rapid and dynamic changes in habitat under a warming climate, continued presence of Bd in the environment for over a decade, and a reduction in one of three anuran species, we document that these anurans continue to breed and survive in this high Andean environment. High variability in anuran encounters across sites and plasticity in these populations across habitats, sites, and years are all factors that could favor repopulation postdecline. Preserving the connectivity of wetlands in the Cordillera Vilcanota is therefore essential in ensuring that anurans continue to breed and adapt as climate change continues to reshape the environment.     

The role of wild grasses in the management of lepidopterous stem-borers on maize in the humid tropics of western Africa

Sites in the humid forest of Cameroon and the derived savanna of Benin were selected to evaluate the effect of planting border rows of wild host plants on lepidopterous stem-borer infestations and on maize yield. Grass species were chosen that in surveys and greenhouse trials were highly attractive to ovipositing female moths but with offspring mortality of close to 100%, thus acting as trap plants. In Cameroon, elephant grass Pennisetum purpureum Moench significantly lowered infestations of Busseola fusca (Fuller), Sesamia calamistis Hampson and Eldana saccharina Walker and increased yields of maize though the differences were not significant during all three cropping seasons. In 1998 in Benin, the only grass tested, Pennisetum polystachion L., significantly increased parasitism of mainly S. calamistis eggs by Telenomus spp. and larvae by Cotesia sesamiae Cameron and reduced numbers of the cob-borer Mussidia nigrivenella Ragonot. In 1999, three grass species; P. polystachion, Sorghum arundinaceum (Desv.) Stapf and Panicum maximum Jacq. were tested. Panicum maximum was the most efficient species for suppressing S. calamistis and M. nigrivenella infestations and enhancing egg and larval parasitism. In the Benin trials, with the exception of M. nigrivenella damage to cobs, the grass species tested had no beneficial effect on yield because pest densities were too low and also rodent damage to maize was enhanced with grasses in the vicinity of the crop. By contrast, stand losses due to Fusarium verticillioides Sacc. (Nirenberg), were significantly reduced by border rows of grasses.     

Urea is not a universal cryoprotectant among hibernating anurans: Evidence from the freeze-tolerant boreal chorus frog (Pseudacris maculata)

Freeze-tolerant organisms accumulate a diversity of low molecular weight compounds to combat negative effects of ice formation. Previous studies of anuran freeze tolerance have implicated urea as a cryoprotectant in the wood frog (Lithobates sylvatica). However, a cryoprotective role for urea has been identified only for wood frogs, though urea accumulation is an evolutionarily conserved mechanism for coping with osmotic stress in amphibians. To identify whether multiple solutes are involved in freezing tolerance in the boreal chorus frog (Pseudacris maculata), we examined seasonal and freezing-induced variation in several potential cryoprotectants. We further tested for a cryoprotective role for urea by comparing survival and recovery from freezing in control and urea-loaded chorus frogs. Tissue levels of glucose, urea, and glycerol did not vary significantly among seasons for heart, liver, or leg muscle. Furthermore, no changes in urea or glycerol levels were detected with exposure to freezing temperatures in these tissues. Urea-loading increased tissue urea concentrations, but failed to enhance freezing survival or facilitate recovery from freezing in chorus frogs in this study, suggesting little role for urea as a natural cryoprotectant in this species. These data suggest that urea may not universally serve as a primary cryoprotectant among freeze-tolerant, terrestrially hibernating anurans.     

Frogs in the spotlight: a 16‐year survey of native frogs and invasive toads on a floodplain in tropical Australia

Although widespread declines in anuran populations have attracted considerable concern, the stochastic demographics of these animals make it difficult to detect consistent trends against a background of spatial and temporal variation. To identify long‐term trends, we need datasets gathered over long time periods, especially from tropical areas where anuran biodiversity is highest. We conducted road surveys of four anurans in the Australian wet–dry tropics on 4637 nights over a 16‐year period. Our surveys spanned the arrival of invasive cane toads (Rhinella marina), allowing us to assess the invader's impact on native anuran populations. Our counts demonstrate abrupt and asynchronous shifts in abundance and species composition from one year to the next, not clearly linked to rainfall patterns. Typically, periods of decline in numbers of a species were limited to 1–2 years and were followed by 1‐ to 2‐year periods of increase. No taxa showed consistent declines over time, although trajectories for some species showed significant perturbations coincident with the arrival of toads. None of the four focal frog species was less common at the end of the study than at the beginning, and three of the species reached peak abundances after toad arrival. Survey counts of cane toads increased rapidly during the initial stage of invasion but have subsequently declined and fluctuated. Distinguishing consistent declines versus stochastic fluctuations in anuran populations requires extensive time‐series analysis, coupled with an understanding of the shifts expected under local climatic conditions. This is especially pertinent when assessing impacts of specific perturbations such as invasive species.     

Larval Tolerance to Salinity in Three Species of Australian Anuran: An Indication of Saline Specialisation in Litoria aurea

Recent anthropogenic influences on freshwater habitats are forcing anuran populations to rapidly adapt to high magnitude changes in environmental conditions or face local extinction. We examined the effects of ecologically relevant elevated salinity levels on larval growth, metamorphosis and survival of three species of Australian anuran; the spotted marsh frog (Limnodynastes tasmaniensis), the painted burrowing frog (Neobatrachus sudelli) and the green and golden bell frog (Litoria aurea), in order to better understand the responses of these animals to environmental change. Elevated salinity (16% seawater) negatively impacted on the survival of L. tasmaniensis (35% survival) and N sudelli (0% survival), while reduced salinity had a negative impact on L. aurea. (16% seawater: 85% survival; 0.4% seawater: 35% survival). L. aurea tadpoles survived in salinities much higher than previously reported for this species, indicating the potential for inter-populations differences in salinity tolerance. In L. tasmaniensis and L. aurea, development to metamorphosis was fastest in low and high salinity treatments suggesting it is advantageous for tadpoles to invest energy in development in both highly favourable and developmentally challenging environments. We propose that this response might either maximise potential lifetime fecundity when tadpoles experience favourable environments, or, facilitate a more rapid escape from pond environments where there is a reduced probability of survival.     

Species and sex biases in ectoparasitism of dragonflies by mites

An important problem in understanding the evolution of parasite host range is determining the extent to which parasite fitness varies among host species and the factors affecting that fitness variation. We present a detailed investigation on the patterns of host use and successful parasitism of two dragonfly species by the ectoparasitic water mite, Limnochares americana Lundblad. In our field surveys, we found both species biases and sex biases in parasitism by mites, which appear explained by differences in exposure. Differential habitat use by dragonflies helped explain male biases in parasitism in both host species, but was not useful in explaining species biases in parasitism. Species biases in parasitism may be explained by more subtle variation in habitat use not explored in this study, or perhaps by differences in timing of emergence, as we found for the two dragonfly species. Despite species differences in parasitism in nature, we found that mites attached equally successfully to both dragonfly species during experimental infestations. However, mites failed to engorge more often on the dragonfly species less often used as a host in nature. This host species also was more likely to have dead mites in natural infestations as compared to the other host species, which was more often and more heavily parasitized. Our results are consistent with previous research suggesting parasites are less successful on less often used hosts. Such research has implications for understanding determinants of host range for animal parasites.     

Anuran gender identification by fecal steroid analysis

This study tested the hypothesis that steroid hormone metabolites can be measured in anuran feces and their concentrations used to identify the sex of adults. Fecal samples from American toads, Bufo americanus, and boreal toads, B. boreas boreas, were extracted using ethyl acetate, and the concentrations of estradiol, progesterone and testosterone metabolites were measured by enzyme immunoassays with antibodies commonly used to evaluate steroid hormone concentrations in mammalian species. In American toads, mean testosterone metabolite concentrations (P<0.05) between males (224.3±15.5 ng/g feces) and females (80.7±10.6 ng/g), but estradiol and progesterone metabolite concentrations did not. In contrast, estradiol immunoreactivity differed (P<0.05) between male (19.0±1.8 ng/g) and female (48.3±6.3 ng/g) boreal toads. Progesterone and testosterone metabolite concentrations did not differ. Fecal hormone metabolite analysis offers a promising noninvasive approach to gender identification in anuran amphibians. However, the group of metabolites differentiating gender may not be consistent among species. Zoo Biol 0:1–12, 2005. © 2005 Wiley‐Liss, Inc.