Effects of salinity on multiplication and transmission of an intertidal trematode parasite

Salinity levels vary spatially in coastal areas, depending on proximity to freshwater sources, and may also be slowly decreasing as a result of anthropogenic climatic changes. The impact of salinity on host–parasite interactions is potentially a key regulator of transmission processes in intertidal areas, where trematodes are extremely common parasites of invertebrates and vertebrates. We investigated experimentally the effects of long-term exposure to decreased salinity levels on output of infective stages (cercariae) and their transmission success in the trematode Philophthalmus sp. This parasite uses the snail Zeacumantus subcarinatus as intermediate host, in which it asexually produces cercariae. After leaving the snail, cercariae encyst externally on hard substrates to await accidental ingestion by shorebirds, which serve as definitive hosts. We found that at reduced salinities (25 or 30 psu), the cercarial output of the parasite was lower, the time taken by cercariae to encyst was longer, fewer cercariae successfully encysted and encysted parasites had lower long-term survival than at normal seawater salinity (35 psu). The strong effect of salinity on the replication and transmission of this parasite suggests that there may be sources and sinks of transmission to birds along coastal areas, depending on local salinity conditions. Also, unless it evolves to adapt to changing conditions, the predicted reduction in salinity as a consequence of climate change may have negative impact on the parasite’s abundance.     

Differential effects of temperature variability on the transmission of a marine parasite

Temperature variability is particularly pronounced in intertidal systems. The importance of considering this variability has been increasingly recognised, especially in the context of climate change and disease dynamics. Here, we investigated the effects of temperature variability on the transmission of the intertidal trematode Maritrema novaezealandensis. The experimental treatments were 15 °C (control), 15 + 5 °C daily, 15 + 10 °C every second day, 15 + 15 °C every third day (overall equal thermal loading), and a heat wave treatment (15 + 10 °C daily). Daily 6 h incubations were carried out corresponding to daytime low tides over a 12-day period. Effects on output of transmission stages (cercariae) from infected Zeacumantus subcarinatus snail hosts and transmission success of cercariae to Paracalliope novizealandiae amphipod hosts were quantified, as well as the survival of amphipods. Results showed differential effects on output and transmission success. The number of cercariae emerging was similar for treatments with equal thermal loading, but was substantially increased in the heat wave treatment. Transmission success was highest and comparable for the treatments with regular daily temperature increases (i.e. 15 + 5 °C and heat wave), compared to other treatments. Amphipod survival was not affected by temperature treatment directly, but by the number of parasites infecting an amphipod, as well as amphipod sex. These results demonstrate that cercarial output depends mostly on total thermal loading, whereas successful infection of amphipods is determined by total time above 15 °C. Repeated exposure to ~25 °C, as expected under a heat wave scenario, therefore increases both transmission pressure and success, and hence, the risk of parasite-induced mortality in amphipods.     

Parasite competition hidden by correlated coinfection: using surveys and experiments to understand parasite interactions

Within most free-living species exists a cryptic community of interacting parasites. By combining multiscale field data with manipulative experiments, we evaluated patterns of parasite coinfection in amphibian hosts and their underlying mechanisms. Surveys of 86 wetlands and 1273 hosts revealed positive correlations between two pathogenic trematodes (Ribeiroia ondatrae and Echinostoma trivolvis) both between wetlands and within individual hosts. In infection and coinfection experiments, Ribeiroia caused greater pathology than Echinostoma, including high host mortality (24%) and severe limb malformations (75%). No interactive effects were noted for host pathology, but both parasites decreased the per capita persistence of one another by 17-36%. Thus, in spite of consistently positive associations from field data, these parasites negatively affected the persistence of one another, likely via cross immunity (apparent competition). These findings underscore the danger of inferring parasite interactions from coinfection patterns and emphasize the potential disconnect between within-host processes (e.g., competition) and between-host processes (e.g., exposure and transmission). Here, correlated coinfections likely resulted from similarities in the parasites' host requirements and heterogeneity in host susceptibility or exposure. Understanding complex interactions among parasites depends critically on the scale under consideration, highlighting the importance of combining coinfection field studies with mechanistic experiments.     

Controls of spatial variation in the prevalence of trematode parasites infecting a marine snail

Geographic variability in abundance can be driven by multiple physical and biological factors operating at multiple scales. To understand the determinants of larval trematode prevalence within populations of the marine snail host Littorina littorea, we quantified many physical and biological variables at 28 New England intertidal sites. A hierarchical, mixed-effects model identified the abundance of gulls (the final hosts and dispersive agents of infective trematode stages) and snail size (a proxy for time of exposure) as the primary factors associated with trematode prevalence. The predominant influence of these variables coupled with routinely low infection rates (21 of the 28 populations exhibited prevalence <12%) suggest broad-scale recruitment limitation of trematodes. Although infection rates were spatially variable, formal analyses detected no regional spatial gradients in either trematode prevalence or independent environmental variables. Trematode prevalence appears to be predominantly determined by local site characteristics favoring high gull abundance.     

除草剂西玛津对非洲爪蟾生存和性腺发育的影响

为了研究三嗪类除草剂西玛津对两栖动物非洲爪蟾(Xenopus laevis)生存和性腺发育的毒性作用,并且与另一种三嗪类除草剂阿特拉津的毒性进行比较,将非洲爪蟾从46/47阶段开始暴露西玛津和阿特拉津到变态1个月后停止,再饲养2个月后将其解剖,取性腺做形态学和组织学观察.暴露期间,每天记录蝌蚪的生长发育情况和存活率.结果显示,在暴露的第1周内(蝌蚪处于46~50阶段),西玛津可导致蝌蚪死亡率明显升高,随后的时间内西玛津对非洲爪蟾的生存不再有明显影响,但却使发育阶段明显的不整齐.阿特拉津对非洲爪蟾的生存和发育则没有明显影响.西玛津和阿特拉津对非洲爪蟾性腺的总体形态和性别比没有明显影响,然而两种除草剂均在一定程度上导致了睾丸组织学的改变.西玛津可能与阿特拉津一样能够通过雌性化/去雄性化作用影响非洲爪蟾睾丸的发育.     

Interactions of an Insecticide with Larval Density and Predation in Experimental Amphibian Communities

Abstract: This study examines the effects of the short-lived insecticide carbaryl, a neurotoxin, on amphibian communities experiencing natural stresses of competition and predation. Tadpoles of three species (  Woodhouse's toad [ Bufo woodhousii ], gray treefrog [ Hyla versicolor ], and green frog [ Rana clamitans ]), representing a commonly encountered assemblage in Missouri, were reared in outdoor polyethylene pond mesocosms. We determined the effects of initial tadpole density ( low or high), predation (newts [  Notophthalmus viridescens ] absent or present), chemical exposure (0, 3.5, or 7.0 mg /L carbaryl), and their interactions on body mass, larval period, and survival to metamorphosis. Green frogs in high-density ponds did not reach metamorphosis, but metamorphs in low-density ponds and tadpoles in high-density ponds were not significantly affected by treatments or their interaction. Carbaryl reduced survival to metamorphosis in toads and treefrogs and increased mass at metamorphosis in treefrogs. Effects of carbaryl varied with predator environment and initial larval density. Interactions of carbaryl with predator and with density may result in an indirect effect of carbaryl causing increased food resources through the elimination of zooplankton populations that may compete for similar resources. Our results indicate that differences in biotic conditions influenced the potency of carbaryl and that even low concentrations induce changes that may alter community dynamics in ways not predicted from single-factor, laboratory-based studies.     

Gonadal histology and reproductive steroidogenesis in Lithobates pipiens exposed to atrazine

Abstract

Atrazine, an endocrine-disrupting compound and environmental pollutant, can have adverse effects on amphibian reproductive development and function. To determine the effects of atrazine on anuran gonadal development, Lithobates pipiens larvae were exposed from Gosner stage 25 through 1-month post metamorphosis to 0, 2, 20, or 200?μg/L atrazine or estradiol. Exposure to atrazine did not markedly alter tadpole growth, development, gonadal histology, or reproductive steroidogenesis. Testicular ovarian follicles appeared in L. pipiens metamorphs following all treatments, including controls, whereas ovotestes occurred only in positive controls. Testicular ovarian follicles may be a natural ontogenetic occurrence in this species while ovotestes indicate disruption in development. Establishing the normal pattern of reproductive development for anuran species and conducting comparisons in fully sexually differentiated animals is necessary to clarify the influence of endocrine disrupting compounds.     

Atrazine increases ranavirus susceptibility in the tiger salamander, Ambystoma tigrinum.

Pathogenic diseases and environmental contaminants are two of the leading hypotheses for global amphibian declines, yet few studies have examined the influence of contaminants on disease susceptibility. In this study, we examined effects of ecologically relevant doses of atrazine (0, 1.6, 16, and 160 microg/L), sodium nitrate (0, 6.8, 68 mg/L), and their interactions on susceptibility of four laboratory-bred tiger salamander families to Ambystoma tigrinum virus (ATV), a pathogen implicated in global amphibian die-offs. Salamanders were from Arizona populations where a coevolutionary history with ATV is supported, and thus cofactors rather than recent introduction may contribute to disease epizootics. Use of atrazine and nitrogenous fertilizers are ubiquitous; therefore, the impact of these cofactors on disease susceptibility is an important consideration. Atrazine and sodium nitrate significantly decreased peripheral leukocyte levels, suggesting an impact of these contaminants on the immune system. As expected from this result, atrazine significantly increased susceptibility of larvae to ATV infection. In contrast, nitrate had a marginally significant main effect and significantly decreased infection rate at the highest level. However, neither atrazine nor sodium nitrate had significant effects on viral copy number per individual. These results suggest that ecologically relevant concentrations of atrazine and nitrates have immunosuppressive effects, and atrazine may contribute to ATV epizootics, raising concerns about the influence of contaminants on diseases in general.     

Road Facilitation of Trematode Infections in Snails of Northern Alaska

Abstract:  Road disturbances can influence wildlife health by spreading disease agents and hosts or by generating environmental conditions that sustain these agent and host populations. I evaluated field patterns of trematode infections in snails inhabiting ponds at varying distances from the Dalton Highway, a wilderness road that intersects northern Alaska. I also assessed the relationships between trematode infections and snail densities and six environmental variables: calcium concentration, aquatic vegetative cover, canopy cover, temperature, pond size, and community structure. Presence of trematode infections and snail density were negatively correlated with distance from the highway. Of the pond characteristics measured, only calcium concentration and vegetation density declined with distance from road. However, neither variable was positively associated with snail density or trematode presence. One potential explanation for observed patterns is that vehicles, road maintenance, or vertebrate vectors attracted to the highway facilitate colonization of snails or trematodes. Emerging disease threats to biological diversity in northern ecosystems highlight the importance of understanding how roads affect disease transmission.     

Effects of chytrid fungus and a glyphosate-based herbicide on survival and growth of wood frogs (Lithobates sylvaticus)

Anthropogenic-derived stressors in the environment, such as contaminants, are increasingly considered important cofactors that may decrease the immune response of amphibians to pathogens. Few studies, however, have integrated amphibian disease and contaminants to test this multiple-stressor hypothesis for amphibian declines. We examined whether exposure to sublethal concentrations of a glyphosate-based herbicide and two strains of the pathogenic chytrid fungus, Batrachochrytrium dendrobatidis (Bd) could: (1) sublethally affect wood frogs (Lithobates sylvaticus) by altering the time to and size at metamorphosis, and (2) directly affect survivability of wood frogs after metamorphosis. Neither Bd strain nor herbicide exposure alone significantly altered growth or time to metamorphosis. The two Bd strains did not differ in their pathogenicity, and both caused mortality in post-metamorphic wood frogs. There was no evidence of an interaction between treatments, indicating a lack of herbicide-induced susceptibility to Bd. However, the trends in our data suggest that exposure of wood frogs to a high concentration of glyphosate-based herbicide may reduce Bd-caused mortality compared to animals exposed to Bd alone. These results exemplify the complexities inherent when populations are coping with multiple stressors. In this case, the perceived stressor, glyphosate-based herbicide, appeared to affect the pathogen more than the host's immune system, relieving the host from disease-caused effects. This suggests caution when invoking multiple stressors as a cause for increased disease susceptibility and indicates that the effects of multiple stressors on disease outcome depend on the interrelationships of stressors to both the pathogen and the host.     

Species diversity reduces parasite infection through cross-generational effects on host abundance

With growing interest in the effects of biodiversity on disease, there is a critical need for studies that empirically identify the mechanisms underlying the diversity-disease relationship. Here, we combined wetland surveys of host community structure with mechanistic experiments involving a multi-host parasite to evaluate competing explanations for the dilution effect. Sampling of 320 wetlands in California indicated that snail host communities were strongly nested, with competent hosts for the trematode Ribeiroia ondatrae predominating in low-richness assemblages and unsuitable hosts increasingly present in more diverse communities. Moreover, competent host density was negatively associated with increases in snail species richness. These patterns in host community assembly support a key prerequisite underlying the dilution effect. Results of multigenerational mesocosm experiments designed to mimic field-observed community assemblages allowed us to evaluate the relative importance of host density and diversity in influencing parasite infection success. Increases in snail species richness (from one to four species) had sharply negative effects on the density of infected hosts (-90% reduction). However, this effect was indirect; competition associated with non-host species led to a 95% reduction in host density (susceptible host regulation), owing primarily to a reduction in host reproduction. Among susceptible hosts, there were no differences in infection prevalence as a function of community structure, indicating a lack of support for a direct effect of diversity on infection (encounter reduction). In monospecific conditions, higher initial host densities increased infection among adult hosts; however, compensatory reproduction in the low-density treatments equalized the final number of infected hosts by the next generation, underscoring the relevance of multigenerational studies in understanding the dilution effect. These findings highlight the role of interspecific competition in mediating the relationship between species richness and parasite infection and emphasize the importance of field-informed experimental research in understanding mechanisms underlying the diversity-disease relationship.     

Adding infection to injury: synergistic effects of predation and parasitism on amphibian malformations

We explored the importance of interactions between parasite infection and predation in driving an emerging phenomenon of conservation importance: amphibian limb malformations. We suggest that injury resulting from intraspecific predation in combination with trematode infection contributes to the frequency and severity of malformations in salamanders. By integrating field surveys and experiments, we evaluated the individual and combined effects of conspecific attack and parasite (Ribeiroia ondatrae) infection on limb development of long-toed salamanders (Ambystoma macrodactylum). In the absence of Ribeiroia, abnormalities involved missing digits, feet, or limbs and were similar to those produced by cannibalistic attack in experimental trials. At field sites that supported Ribeiroia, malformations were dominated by extra limbs and digits. Correspondingly, laboratory exposure of larval salamanders to Ribeiroia cercariae over a 30-day period induced high frequencies of malformations, including extra digits, extra limbs, cutaneous fusion, and micromelia. However, salamander limbs exposed to both injury and infection exhibited 3-5 times more abnormalities than those exposed to either factor alone. Infection also caused significant delays in limb regeneration and time-to-metamorphosis. Taken together, these results help to explain malformation patterns observed in natural salamander populations while emphasizing the importance of interactions between parasitism and predation in driving disease.     

Nickel induced changes on some aspects of protein metabolism in the tissues of Pila globosa

Some aspects of protein metabolism were studied in foot, hepatopancreas and mantle tissues of snail, Pila globosa on exposure to lethal concentration for 2 days (336.7 mg/L) and sublethal concentration (67.34 mg/L) of nickel for 1, 5 and 10 days. Total, structural and soluble proteins decreased significantly and to continence, this the levels of amino acids and protease activity increased in all the tissues of snail at all time points examined. Activities of AAT (Aspartate aminotransferase) and AlAT (Alanine aminotransferase) showed contrasting trends of inhibition and elevation during lethal and sublethal concentrations of nickel treatment. GDH (Glutamate dehydrogenase) activity was increased in all the tissues with increase in exposure time. Level of ammonia decreased in snails at sublethal concentration, but increment was observed in lethal concentration along with increased urea content. Under lethal and sublethal exposures, the changes in all the parameters were more pronounced in hepatopancreas followed by foot and mantle. At most instances, snails in the lethal medium were affected more compared to sublethal concentration.     

Fecundity of the freshwater fish, Notopterus notopterus (Pallas) in natural and heavy metal contaminated water

The knowledge of fecundity of fish from a specific aquatic body is extremely important in the successful management and exploitation of its fishery. In the present investigation the fecundity of the freshwater fish, Notopterus notopterus was studied in fish collected from a natural aquatic body (Sirnoor nala) near Gulbarga and also in fish exposed to some heavy metal contamination (HgCl2, CdCl2 and their combination) at sublethal concentration for 15 days in the laboratory. The mathematical relationship between fecundity and total length, body weight, ovary length and ovary weight were determined in both unexposed and exposed fish. The fish, N. notopterus has bigger oocytes and are few in number. Studies in the fish exposed to heavy metals indicate that significant reduction in these parameters after exposure to heavy metals at sublethal concentration was noticed. The fecundity has straight line relationship with total length, body weight, ovary length and ovary weight in control fish which did not alter after heavy metal exposure. This study provides the viability of species in only specific environment.     

Relations between Conspecific Density and Effects of Ultraviolet‐B Radiation on Tadpole Size in the Striped Marsh Frog

Global increases in ultraviolet‐B radiation (UVBR) associated with stratospheric ozone depletion are potentially contributing to the decline of numerous amphibian species around the world. Exposure to UVBR alone reduces survival and induces a range of sublethal effects in embryonic and larval amphibians. When additional environmental stressors are present, UVBR can have compounding negative effects. Thus, examination of the effects of UVBR in the absence of other stressors may substantially underestimate its potential to affect amphibians in natural habitats. We examined the independent and interactive effects of increased UVBR and high conspecific density would have embryonic and larval striped marsh frogs (Limnodynastes peronii). We exposed individuals to a factorial combination of low and high UVBR levels and low, medium, and high densities of striped marsh frog tadpoles. The response variables were time to hatching, hatching success, posthatch survival, burst‐swimming performance of tadpoles (maximum instantaneous swim speed following an escape response), and size and morphology of tadpoles. Consistent with results of previous studies, we found that exposure to UVBR alone increased the time to hatching of embryos and reduced the burst‐swimming performance and size of tadpoles. Similarly, increasing conspecific density increased the time to hatching of embryos and reduced the size of tadpoles, but had no effect on burst‐swimming performance. The negative effect of UVBR on tadpole size was not apparent at high densities of tadpoles. This result suggests that tadpoles living at higher densities may invest relatively less energy in growth and thus have more energy to repair UVBR‐induced damage. Lower densities of conspecifics increased the negative effects of UVBR on developing amphibians. Thus, low‐density populations, which may include declining populations, may be particularly susceptible to the detrimental effects of increased UVBR and thus may be driven toward extinction faster than might be expected on the basis of results from single‐factor studies. Relaciones entre la Densidad Conespecífica y los Efectos de la Radiación Ultravioleta‐B sobre el Tamaño de Renacuajos de Limnodynastes peronei     

Variation in Pesticide Tolerance of Tadpoles among and within Species of Ranidae and Patterns of Amphibian Decline

Abstract: There is significant variation among and within amphibian species with respect to reports of population decline; declining species are often found in environments that are physiograpically similar to environments where the same species is thriving. Because variability exists among organisms in their sensitivity to environmental stressors, it is important to determine the degree of this variation when undertaking conservation efforts. We conducted both lethal (time-to-death) and sublethal (activity change) assays to determine the degree of variation in the sensitivity of tadpoles to a pesticide, carbaryl, at three hierarchical levels: among ranid species, among several populations of a single ranid species (   Rana sphenocephala ), and within populations of R. sphenocephala . We observed significant variation in time to death among the nine ranid species and among the 10 R. sphenocephala populations we tested. Four out of eight R. sphenocephala populations exhibited significantly different times to death among families. The magnitude of the activity change in response to exposure to sublethal carbaryl levels was significantly different among species and within R. sphenocephala populations. Chemical contamination, at lethal or sublethal levels, can alter natural regulatory processes such as juvenile recruitment in amphibian populations and should be considered a contributing cause of declines in amphibian populations.     

Parasite transmission in complex communities: predators and alternative hosts alter pathogenic infections in amphibians

While often studied in isolation, host-parasite interactions are typically embedded within complex communities. Other community members, including predators and alternative hosts, can therefore alter parasite transmission (e.g., the dilution effect), yet few studies have experimentally evaluated more than one such mechanism. Here, we used data from natural wetlands to design experiments investigating how alternative hosts and predators of parasites mediate trematode (Ribeiroia ondatrae) infection in a focal amphibian host (Pseudacris regilla). In short-term predation bioassays involving mollusks, zooplankton, fish, larval insects, or newts, four of seven tested species removed 62-93% of infectious stages. In transmission experiments, damselfly nymphs (predators) and newt larvae (alternative hosts) reduced infection in P. regilla tadpoles by -50%, whereas mosquitofish (potential predators and alternative hosts) did not significantly influence transmission. Additional bioassays indicated that predators consumed parasites even in the presence of alternative prey. In natural wetlands, newts had similar infection intensities as P. regilla, suggesting that they commonly function as alternative hosts despite their unpalatability to downstream hosts, whereas mosquitofish had substantially lower infection intensities and are unlikely to function as hosts. These results underscore the importance of studying host-parasite interactions in complex communities and of broadly linking research on predation, biodiversity loss, and infectious diseases.     

Consequences of Embryonic UV-B Exposure for Embryos and Tadpoles of the Plains Leopard Frog

Abstract: In organisms with complex life cycles, understanding the effects of environmental change requires investigating the possibility that the effect of such change on one stage will have consequences for other stages. We investigated the effects of embryonic exposure to UV-B radiation on embryo and tadpole development of the plains leopard frog (   Rana blairi ). Embryos were exposed to sunlight filtered to provide two levels of UV-B exposure (e.g., at 310 nm: high exposure is approximately 84% transmittance and low exposure is approximately 58% transmittance). Hatching success of embryos exposed to different UV-B levels did not differ. Tadpoles resulting from exposed embryos were raised at three densities. Growth and development were slower in tadpoles exposed to higher UV-B levels as embryos. Density negatively affected growth and development. Survivorship did not differ among UV-B or density treatments. Our results suggest that even if lethal effects are not exhibited at one stage, sublethal effects may manifest themselves at other stages, possibly affecting the long-term success of exposed individuals and populations.     

Digenean trematodes infecting the tropical abalone <Emphasis Type="Italic">Haliotis asinina</Emphasis> have species-specific cercarial emergence patterns that follow daily or semilunar spawning cycles

Approximately 1–2% of the tropical abalone Haliotis asinina inhabiting Heron Island Reef are infected with opecoelid digeneans. These largely inhabit the haemocoel surrounding the cerebral ganglia and digestive gland–gonad complex, and infected abalone typically have significantly reduced or ablated gonads. Observations of infected abalone reveal two distinct cercarial emergence patterns, one which correlates tightly with the abalone’s highly regular and synchronous fortnightly spawning cycle, and the other which occurs in a circadian pattern. The former appears to be a novel emergence strategy not previously observed in digeneans. While the cercariae in all abalone are morphologically indistinguishable, comparison of sequences from the internal transcribed spacer 2 (ITS 2) region of the ribosomal DNA reveals a 5.7% difference between cercariae displaying different emergence patterns, indicating these are two distinct species that probably belong to the same genus. The ITS 2 sequences of the species with the daily emergence pattern are identical to that of an undescribed adult opecoelid from the gut of the barramundi cod, Cromileptes altivelis. Combined molecular, morphological and emergence data suggest that while these opecoelid cercariae use the same first intermediate host and are closely related species—members of the genus Allopodocotyle—they fill different ecological niches that are likely to include different definitive hosts.     

Local variation of within-host clonal diversity coupled with genetic homogeneity in a marine trematode

Despite their ubiquity and importance to intertidal ecosystems, information is currently lacking regarding the genetic diversity of trematode parasites within coastal organisms and the distribution of their genetic variation among intertidal habitats. In this study, we quantified the clonal diversity of the coastal marine trematode Maritrema novaezealandensis within Zeacumantus subcarinatus snail hosts from three coastal bays in Otago Harbour, New Zealand, using five microsatellite loci to determine if differences exist in the frequency of occurrence of multi-clone infections. In addition, we examined gene flow among M. novaezealandensis collected from the three bays. The frequency of mixed-clone infections varied fourfold among bays and no genetic differentiation was detected among intertidal bays. Across the coastal bays studied, M. novaezealandensis comprises a single population that is potentially infecting multiple Z. subcarinatus populations with varying life history traits.