Suburbanization Increases Echinostome Infection in Green Frogs and Snails.

An important contribution to infectious disease emergence in wildlife is environmental degradation driven by pollution, habitat fragmentation, and eutrophication. Amphibians are a wildlife group that is particularly sensitive to land use change, infectious diseases, and their interactions. Residential suburban land use is now a dominant, and increasing, form of land cover in the USA and globally, contributing to increased pollutant and nutrient loading in freshwater systems. We examined how suburbanization affects the infection of green frog (Rana clamitans) tadpoles and metamorphs by parasitic flatworms (Echinostoma spp.) through the alteration of landscapes surrounding ponds and concomitant changes in water quality. Using sixteen small ponds along a forest-suburban land use gradient, we assessed how the extent of suburban land use surrounding ponds influenced echinostome infection in both primary snail and secondary frog hosts. Our results show that the degree of suburbanization and concurrent chemical loading are positively associated with the presence and burden of echinostome infection in both host populations. This work contributes to a broader understanding of how land use mediates wildlife parasitism and shows how human activities at the household scale can have similar consequences for wildlife health as seemingly more intensive land uses like agriculture or urbanization.

Mass Mortality of Green Frog ( Rana clamitans) Tadpoles in Wisconsin, USA, Associated with Severe Infection with the Pathogenic Perkinsea Clade.

We documented mortality of green frog ( Rana clamitans) tadpoles in Wisconsin, US, attributed to severe Perkinsea infection. Final diagnosis was determined by histopathology. followed by molecular detection of pathogenic Perkinsea clade (PPC) of frogs in the liver. To our knowledge, this represents the first detection of PPC in the midwestern US.

Catastrophic collapse of the larval trematode component community in Charlie's Pond (North Carolina).

In 1984, work on the parasite population and community ecology in the pulmonate snail, Helisoma anceps , was initiated in Charlie's Pond (North Carolina). Similar research on Physa gyrina was started in 1986. When study in the pond began in 1984, 8 species of larval trematodes were being shed from Hel. anceps. By far, the dominant species was Halipegus occidualis , with prevalences generally ∼60%, except during midsummer, when older snails were dying. For the other 7 trematode species being shed, prevalences were consistently less than 4%. By 2006, 18 species had been identified in Hel. anceps at one time or another. In 1986, Hal. eccentricus was discovered in P. gyrina , with a prevalence of ∼49%. Through 2006, 7 trematodes were found to be shedding cercariae from P. gyrina . Halipegus eccentricus disappeared from the pond in 1998. From March through November of 2012 and 2013, 1,292 Hel. anceps and 716 P. gyrina were collected, using collection protocols that were identical to those used from 1984 through 2006. In 2012, 5 trematode species, including Hal. occidualis, were present in Hel. anceps at one time or another. During the last part of the 2012 collecting season cercariae of just 2 species were being shed from Hel. anceps (and 1 from P. gyrina ). In 2013, only cercariae of Haematoloechus longiplexus and Uvulifer ambloplitis were observed from Hel. anceps. The latter species was lost by 2014, and an echinostome was present (2.1%); a single snail was infected with Haem. longiplexus. Four species were being shed from P. gyrina , i.e., Echinoparyphium sp. (7.9%), Glypthelmins sp. (1.5%), Plagiorchis sp. (4.9%), and Posthodiplostomum sp. (7.4%). Rarefaction curves were generated for Hel. anceps shedding in 1984, 1988, 1989, 2002, 2006, and August of 2014. The data clearly indicate that species diversity was constantly declining over the 31-yr period. We did not include P. gyrina in the analysis since data for this snail species were not acquired until 1991-1992. At present, we have no definitive explanation for the decrease in diversity, although circumstantial evidence suggests that it might be related to periodic declines in water level that negatively affected the colonization and maintenance of emergent vegetation within the pond.

Host density and competency determine the effects of host diversity on trematode parasite infection.

Variation in host species composition can dramatically alter parasite transmission in natural communities. Whether diverse host communities dilute or amplify parasite transmission is thought to depend critically on species traits, particularly on how hosts affect each other's densities, and their relative competency as hosts. Here we studied a community of potential hosts and/or decoys (i.e. non-competent hosts) for two trematode parasite species, Echinostoma trivolvis and Ribeiroia ondatrae, which commonly infect wildlife across North America. We manipulated the density of a focal host (green frog tadpoles, Rana clamitans), in concert with manipulating the diversity of alternative species, to simulate communities where alternative species either (1) replace the focal host species so that the total number of individuals remains constant (substitution) or (2) add to total host density (addition). For E. trivolvis, we found that total parasite transmission remained roughly equal (or perhaps decreased slightly) when alternative species replaced focal host individuals, but parasite transmission was higher when alternative species were added to a community without replacing focal host individuals. Given the alternative species were roughly equal in competency, these results are consistent with current theory. Remarkably, both total tadpole and per-capita tadpole infection intensity by E. trivolvis increased with increasing intraspecific host density. For R. ondatrae, alternative species did not function as effective decoys or hosts for parasite infective stages, and the diversity and density treatments did not produce clear changes in parasite transmission, although high tank to tank variation in R. ondatrae infection could have obscured patterns.

The complete mitochondrial genome sequence of Hepatozoon catesbianae (Apicomplexa: Coccidia: Adeleorina), a blood parasite of the green frog, Lithobates (formerly Rana) clamitans.

A complete mitochondrial genome for the blood parasite Hepatozoon catesbianae (Alveolata; Apicomplexa; Coccidia; Adeleorina; Hepatozoidae) was obtained through PCR amplification and direct sequencing of resulting PCR products. The mitochondrial genome of H. catesbianae is 6,397 bp in length and contains 3 protein-coding genes (cytochrome c oxidase subunit I [COI]; cytochrome c oxidase subunit III [COIII]; and cytochrome B [CytB]). Sequence similarities to previously published mitochondrial genomes of other apicomplexan parasites permitted annotation of 23 putative rDNA fragments in the mitochondrial genome of H. catesbianae, 14 large subunit rDNA fragments, and 9 small subunit rDNA fragments. Sequences corresponding to rDNA fragments RNA5, RNA8, RNA11, and RNA19 of Plasmodium falciparum were not identified in the mitrochondrial genome sequence of H. catesbianae. Although the presence of 3 protein-coding regions and numerous putative rDNA fragments is a feature typical for apicomplexan mitochondrial genomes, the mitochondrial genome of H. catesbianae possesses a structure and gene organization that is distinct among the Apicomplexa. This is the first complete mitochondrial genome sequence obtained from any apicomplexan parasite in the suborder Adeleorina.

Fecundity reduction in the second gonotrophic cycle of Culex pipiens infected with the apicomplexan blood parasite, Hepatozoon sipedon.

Fecundity reduction is a well-recognized phenomenon of parasite infection in insects. Reduced production of eggs might increase longevity of a host and release nutrients to both host and parasite that would otherwise be used for oogenesis. The objective of this study was to assess effects on fecundity caused by Hepatozoon sipedon, an apicomplexan blood parasite of snakes, in its invertebrate host, the mosquito Culex pipiens. In the first gonotrophic cycle, the mean number of eggs laid by mosquitoes infected with H. sipedon did not differ significantly from those laid by uninfected mosquitoes. However, in the second gonotrophic cycle infected mosquitoes laid significantly fewer eggs than did uninfected mosquitoes, and fecundity was reduced by 100% in mosquitoes with parasite burdens of more than 60 oocysts. There was a significant negative correlation between parasite burden, or the number of oocysts, and the number of eggs produced in the second gonotrophic cycle. Significantly fewer viable larvae hatched from eggs laid by infected compared to uninfected mosquitoes in the second gonotrophic cycle. These data indicate that fecundity reduction occurs in this system, although the physiological mechanisms driving this phenotype are not yet known.

The role of gamont entry into erythrocytes in the specificity of Hepatozoon species (Apicomplexa: Adeleida) for their frog hosts.

Hepatozoon species are apicomplexan parasites that infect blood cells and viscera of terrestrial vertebrates. One species, Hepatozoon clamatae, primarily infects green frogs, Rana clamitans , whereas another, Hepatozoon catesbianae, primarily infects bullfrogs, Rana catesbeiana , although both species of parasite are capable of infecting either species of frog. The aim of this study was to determine whether the basis for this partial host specificity is manifested at the gamont, or intraerythrocytic, stage of the parasite's life cycle. Blood was drawn from infected frogs and treated in vitro with a saline solution to induce intracellular gamonts to emerge from host erythrocytes. This treated blood was added to in vitro samples of uninfected blood of green frogs and bullfrogs. After 1 hr, samples were analyzed to determine the level of re-entry of the parasites into uninfected erythrocytes. Results obtained using multiple combinations of donor and recipient frogs indicate that extracellular gamonts of both parasite species do not exhibit preference for erythrocytes of 1 frog species over those of another. These results suggest that the basis for the observed host specificity is not determined at the gamont stage and is more likely dependent on another stage in the parasite life cycle.

Synergistic effects of predators and trematode parasites on larval green frog (Rana clamitans) survival.

Parasites and predators can have complex, nonadditive effects on a shared group of victims, which can have important consequences for population dynamics. In particular, parasites can alter host traits that influence predation risk, and predators can have nonconsumptive effects on prey traits which influence susceptibility (i.e., infection intensity and tolerance) to parasites. Here, we examined the combined effects of trematode parasites (Digenea: Echinostomatidae) and odonate (Anax) predators on the survival of larval green frogs (Rana clamitans). First, in a large-scale mesocosm experiment, we manipulated the presence or absence of parasites in combination with the presence of no predator, caged predators, or free predators, and measured survival, traits, and infection. Parasites, caged predators, and free predators decreased survival, and we found a strong negative synergistic effect of parasites in combination with free predators on survival. Importantly, we then examined the potential mechanisms that explain the observed synergistic effect of parasites and predators in a series of follow-up experiments. Results of the follow-up experiments suggest that increased predation susceptibility due to elevated activity levels in the presence of free-swimming parasite infective stages (i.e., an avoidance response) is the most likely mechanism responsible for the observed synergism. These results suggest a potential trade-off in susceptibility to parasites and predators, which can drive nonadditive effects that may have important consequences for natural enemy interactions in natural populations and amphibian conservation.