Endohelminth fauna of the marsh frog Rana ridibunda from Lake Hazar, Turkey

In this study, 236 marsh frogs Rana ridibunda collected from Lake Hazar (Elazig, Turkey) at 15 d intervals between March 2001 and February 2002 were examined for endohelminths; of these, 148 (62.71%) frogs were found to be infected with helminths. In total, 9 helminth species (3 trematodes, 5 nematodes and 1 acanthocephalan) were identified. We observed Gorgoderina vitelliloba (prevalence 2.97%) in the urinary bladder, Haematoloechus variegatus (4.66%) and Rhabdias bufonis (8.90%) in the lung, Pleurogenoides medians (1.69%), Oswaldocruzia filiformis (3.81 %) and Acanthocephalus ranae (26.27 %) in the small intestine, Neoxysomatium brevicaudatum (16.95%) and Cosmocercoides sp. (3.39%) in the large intestine, and Eustrongylides excisus (14.41%) in the body cavity and on,the stomach. No helminth was found in the spleen, kidney, gall bladder, liver, heart or muscle. Of the 9 helminth species identified, Acanthocephalus ranae (26.27 %) had the highest prevalence and abundance and Oswaldocruzia filiformis (8.33+/-4.09) had the highest mean intensity.     

Helminth communities of green frogs Rana clamitans Latreille, from southwestern Michigan.

A total of 239 green frogs Rana clamitans, collected between June 3 and August 27, 1998 from 6 locations in southwestern Michigan, was examined for helminths. Of the 26 helminth taxa found, the larval cestode Mesocestoides sp. had the highest mean intensity, followed by the larval trematode Fibricola sp. Of the helminths that mature in frogs, Haematoloechus varioplexus had the highest prevalence and Gorgodera amplicava had the highest mean intensity. Frogs from 118th Pond had the highest species richness (20), mean helminth species richness (5.2), and mean helminth abundance (153.7). Frogs from Constantine East had the highest mean helminth species diversity (0.8778) and evenness (0.6033), followed by frogs from 118th Pond. In all comparisons of mean helminth community species richness, abundance, diversity, and evenness, adult frogs had significantly higher or higher values than did juveniles at each location. Jaccard's coefficients of similarity for the helminth communities for location pairs ranged from 0.545 to 0.823. Nine and 2 core helminth taxa occurred at the local and regional levels, respectively. The differences in several helminth community measures in green frogs among locations stress the importance of local ecological conditions on helminth community structure.     

Helminth community structure of sympatric eastern American toad, Bufo americanus americanus, northern leopard frog, Rana pipiens, and blue-spotted salamander, Ambystoma laterale, from southeastern Wisconsin

One-hundred twelve amphibians, including 51 blue-spotted salamanders, Ambystoma laterale, 30 eastern American toads, Bufo americanus americanus, and 31 northern leopard frogs, Rana pipiens, were collected during April-October 1996 from Waukesha County, Wisconsin and examined for helminth parasites. The helminth compound community of this amphibian assemblage consisted of at least 10 species: 9 in American toads, 8 in leopard frogs, and 3 in blue-spotted salamanders. American toads shared 7 species with leopard frogs, and 2 species occurred in all 3 host species. Although there was a high degree of helminth species overlap among these sympatric amphibians, statistically significant differences were found among host species and percent of indirect or direct-life cycle parasites of amphibian species individual component communities (chi2 = 1,015, P < 0.001). American toads had a higher relative abundance of nematodes, 59%, than larval cestodes, 31%, and larval and adult trematodes, 10%, whereas leopard frogs had a higher relative abundance of larval cestodes, 71.3%, and larval and adult trematodes, 25.3%, than nematodes 3.4%. This is related to ecological differences in habitat and dietary preferences between these 2 anuran species. Helminth communities of blue-spotted salamanders were depauperate and were dominated by larval trematodes, 94%, and few nematodes, 6%. Low helminth species richness in this host species is related to this salamander's relatively small host body size, smaller gape size, lower vagility, and more fossorial habitat preference than the other 2 anuran species. Adult leopard frogs and toads had significantly higher mean helminth species richness than metamorphs, but there was no significant difference in mean helminth species richness among adult and metamorph blue-spotted salamanders. Considering adult helminths, the low species richness and low vagility of caudatans as compared with anurans suggest that local factors may be more important in structuring caudatan helminth communities of salamanders than of anuran hosts. Helminth species infecting salamanders may be more clumped in their geographic distribution as compared with anurans, and the role of other hosts and their parasites at the compound community level may be important in structuring helminth communities of salamanders.     

Helminth infracommunities of Rana vaillanti brocchi (Anura: Ranidae) in Los Tuxtlas, Veracruz, Mexico

A total of 76 adult individuals of Rana vaillanti were collected in Laguna Escondida, Los Tuxtlas, Veracruz, Mexico, and their helminth infracommunity structure was determined. Among the 21 helminth taxa collected (10 digeneans, 8 nematodes, and 3 acanthocephalans), the digenean Langeronia macrocirra reached the highest prevalence (64.4%), mean abundance (6.6), and mean intensity (10.4), as well as the highest total number of individuals (499). Only 2 frogs were uninfected, the remainder harbored between 1 and 7 helminth species and 1-102 individuals; mean species richness and abundance were 3.49 +/- 0.22 and 16.1 +/- 16.3, respectively. Langeronia macrocirra dominated in 50.6% of the infracommunities, with relatively low Berger-Parker index values (0.56); for this reason, the evenness was high (0.70 +/- 0.31), and consequently, diversity values are the highest recorded to date in species of Rana. However, patterns of helminth infracommunity richness and diversity were similar to those previously observed in amphibians. This structure is attributed to the feeding habits (between 66.7 and 81% of helminth species parasitizing R. vaillanti enter using the food web dynamics) and low vagility (the remainder species infect by host penetration).     

The physiology of hibernation in Canadian leopard frogs (Rana pipiens) and bullfrogs (Rana catesbeiana)

Canadian northern leopard frogs (Rana pipiens) and bullfrogs (Rana catesbeiana) were acclimated to 3 degrees C and submerged in anoxic (0-5 mmHg) and normoxic (Po(2) approximately 158 mmHg) water. Periodic measurements of blood Po(2), Pco(2), and pH were made on samples taken anaerobically from subsets of each species. Blood plasma was analyzed for [Na(+)], [K(+)], [Cl(-)], [lactate], [glucose], total calcium, total magnesium, and osmolality. Blood hematocrit was determined, and plasma bicarbonate concentration was calculated. Both species died within 4 d of anoxic submergence. Anoxia intolerance would rule out hibernation in mud, which is anoxic. Both species survived long periods of normoxic submergence (R. pipiens, 125 d; R. catesbeiana, 150 d) with minimal changes in acid-base and ionic status. We conclude that ranid frogs require a hibernaculum where the water has a high enough Po(2) to drive cutaneous diffusion, allowing the frogs to extract enough O(2) to maintain aerobic metabolism, but that an ability to tolerate anoxia for several days may still be ecologically meaningful.     

Aegyptianella ranarum sp. n. (Rickettsiales, Anaplasmataceae): ultrastructure and prevalence in frogs from Ontario

Aegyptianella ranarum sp. n. (Rickettsiales, Anaplasmataceae) was recorded from bullfrogs (Rana catesbeiana Shaw), green frogs (Rana clamitans Latreille) and mink frogs (Rana septentrionalis Baird) from five sites in southern Ontario. The rickettsia occurs within membrane-bound vacuoles in the cytoplasm of erythrocytes with up to 120 organisms in mature inclusions. The pattern of replication of A. ranarum in host erythrocytes and its prevalence over a 3-yr period in frogs from Algonquin Park, Ontario are discussed.     

Helminth communities in five species of sympatric amphibians from three adjacent ephemeral ponds in southeastern Wisconsin

Representatives of 5 amphibian species (313 individuals), including eastern American toads (Bufo americanus), wood frogs (Rana sylvatica), spring peepers (Pseudacris crucifer), blue-spotted salamanders (Ambystoma laterale), and central newts (Notophthalmus viridescens louisianensis), were collected from 3 ephemeral ponds during spring 1994, and they were inspected for helminth parasites. The component communities of anurans were more diverse than those of caudates. Infracommunities of all host species were isolationist and depauperate, due mostly to host ectothermy and low vagility. Toad infracommunities were dominated by skin-penetrating nematodes, and they had the highest values of mean total parasite abundance, mean species richness, and overall prevalence. This was likely due to their greater vagility compared with other host species. Infracommunities of wood frogs and blue-spotted salamanders had intermediate values for these measures of parasitism, whereas spring peeper and newt infracommunities had the lowest values. In addition to relative vagility, feeding habits and habitat preference were likely important in helminth community structure. Body size also seemed to play a role because mean wet weight of host species followed the same general trend as values of parasitism. However, effects of size were variable within host species and difficult to separate from other aspects of host ecology.     

Helminth assemblages of the turtle Emydura macquarii (Pleurodira: Chelidae) Queensland, Australia

The helminth fauna of 76 Emydura macquarii from 3 river systems in central and northern Queensland was examined. Eleven species were found, including 2 nematodes, 6 trematodes, 1 aspidogastrean, 1 cestode, and 1 monogenean. Analysis of helminth diversity showed that the Fitzroy and Ross River turtles had communities of comparable diversity, but the helminth communities in Proserpine River turtles were much less diverse. The helminth communities in all localities were dominated by trematodes. Polystomoides australiensis was the most prevalent, being found in 60% of the Ross River turtles, 57% of the Fitzroy River turtles, and 46% of the Proserpine River turtles. Notopronocephalus peekayi was the most abundant species, with mean abundances of 5.9 in the Ross River turtles and 9.8 in the Fitzroy River turtles. Species richness, Simpson's Reciprocal Index, was highest, 4.68, for the Ross River helminth community, Sorensen's Qualitative Index showed 95% similarity between the Ross River and Fitzroy River communities, although Sorensen's Quantitative Index indicated only 35% similarity between the 2 sites. Host feeding patterns are likely the most important factor affecting species richness of the helminth infracommunities, as the majority of helminth species are transmitted by food-web interactions involving intermediate hosts.     

Helminth and leech community structure in tadpoles and caudatan larvae of two amphibian species from Western Nebraska

Currently no comparative studies exist on helminth and leech community structure among sympatric anuran tadpoles and salamander larvae. During June-August 2007-2009, we examined 50 bullfrog tadpoles, Rana catesbeiana , 50 barred tiger salamander larvae, Ambystoma mavortium , and 3 species of snails from Nevens Pond, Keith County, Nebraska for helminth and leech infections. The helminth and leech compound community of this larval amphibian assemblage consisted of at least 7 species, 4 in bullfrog tadpoles and 4 in barred tiger salamander larvae. Bullfrog tadpoles were infected with 2 species of nematodes ( Gyrinicola batrachiensis and Spiroxys sp.) and 2 types of metacercariae ( Telorchis sp. and echinostomatids), whereas barred tiger salamander larva were infected with 1 species of leech ( Placobdella picta ), 2 species of adult trematodes ( Telorchis corti and Halipegus sp.), and 1 species of an unidentified metacercaria. The component community of bullfrog tadpoles was dominated by helminths acquired through active penetration, or incidentally ingested through respiratory currents, or both, whereas the component community of larval salamanders was dominated by helminths acquired through ingestion of intermediate hosts (χ2 = 3,455.00, P < 0.00001). Differences in amphibian larval developmental time (2-3 yr for bullfrog tadpoles versus 2-5 mo for salamander larvae), the ephemeral nature of intermediate hosts in Nevens Pond, and the ability of bullfrog tadpole to eliminate echinostome infections had significant effects on mean helminth species richness among amphibian species and years (t = 12.31, P < 0.0001; t = 2.09, P = 0.04). Differences in herbivorous and carnivorous diet and time to metamorphosis among bullfrog tadpoles and barred tiger salamander larvae were important factors in structuring helminth communities among the larval stages of these 2 sympatric amphibian species, whereas size was important in structuring helminth and leech communities in larval salamanders, but not in bullfrog tadpoles.     

Helminth communities of two green frogs (Rana perezi and Rana saharica) from both shores of the Alboran Sea

The helminth communities of two populations of green frogs from both shores of the Alborón Sea (Western Mediterranean) were studied. Of the 79 frogs examined for helminths, 39 individuals of the species Rano saharica were collected from Bob-Taza (Morocco), and 40 of the species Rona perezi were collected from the Natural Park of the Sierra de Grazalema (Spain). Although the species richness of helminths was identical in the two sampled areas, the differences observed in the structure of the helminth infracommunities were quite important. Statistically, significant differences were found between the species richness and the diversity of the infracommunities of R. perezi female population and the other three studied statistical populations. The helminth component communities of these two green frogs can be considered as depauperate, although their infracommunities present interactive features.     

Effects of the nematode Gyrinicola batrachiensis on development, gut morphology, and fermentation in bullfrog tadpoles (Rana catesbeiana): a novel mutualism

We describe a novel mutualism between bullfrog tadpoles (Rana catesbeiana) and a tadpole-specific gastrointestinal nematode (Gyrinicola batrachiensis). Groups of tadpoles were inoculated with viable or nonviable nematode eggs, and development, morphology, and gut fermentation activity were compared between nematode-infected and uninfected tadpoles. Nematode infection accelerated tadpole development; the mean time to metamorphosis was 16 d shorter and the range of times to metamorphosis was narrower in nematode-infected tadpoles than in uninfected tadpoles. At metamorphosis, infected and uninfected bullfrogs did not differ in body size or condition. Colon width, wet mass of colon contents, and concentrations of most fermentation byproducts (short-chain fatty acids: SCFAs) in the hindgut were greater in infected tadpoles. Furthermore, in vitro fermentation yields for all SCFAs combined were over twice as high in infected tadpoles than in uninfected tadpoles. One explanation for accelerated development in infected tadpoles is the altered hindgut fermentation associated with the nematodes. Energetic contributions of fermentation were estimated to be 20% and 9% of the total daily energy requirement for infected and uninfected tadpoles, respectively. Infection by G. batrachiensis nematodes potentially confers major ecological and evolutionary advantages to R. catesbeiana tadpoles. The mutualism between these species broadens our understanding of the taxonomic diversity and physiological contributions of fermentative gut symbionts and suggests that nematodes inhabiting the gut regions of other ectothermic herbivores might have beneficial effects in those hosts.     

Ichthyophonus-like infection in wild amphibians from Québec, Canada

Myositis associated with infection by Ichthyophonus-like organisms was diagnosed in 35 of 260 (13%) wild amphibians collected in Quebec, Canada, from 1959 to 1964 (n = 30), and 1992 to 1999 (n = 230). Infection was diagnosed in 17 green frogs Rana clamitans, 9 wood frogs R. sylvatica, 4 red-spotted newts Notophthalmus viridescens, 3 bullfrogs R. catesbeiana, 1 spring peeper Pseudacris crucifer, and 1 pickerel frog R. palustris. The spring peeper and one of the bullfrogs were collected in 1964 from the Mont Saint-Hilaire Biosphere Reserve, indicating long-term presence of the organism. Spores of the organisms invaded striated muscle fibers and were associated with variable degrees of granulomatous and eosinophilic inflammation. Infection was considered fatal in 2 green frogs, 1 wood frog, and 1 red-spotted newt. It was considered potentially significant in 3 additional green frogs in which up to 100% of the fibers of some muscles were replaced by spores associated with a severe granulomatous reaction. Ultrastructural features of Ichthyophonus-like spores included a thick trilaminated wall, a paramural cytoplasm, multiple nuclei, oval mitochondria with short tubulo-vesicular cristae and numerous ribosomes. This report represents 4 new host records and shows that ichthyophonosis is enzootic in amphibians from Quebec.     

Latitudinal gradient in the taxonomic composition of parasite communities

Although latitudinal gradients in diversity have been well studied, latitudinal variation in the taxonomic composition of communities has received less attention. Here, we use a large dataset including 950 surveys of helminth endoparasite communities in 650 species of vertebrate hosts to test for latitudinal changes in the relative contributions of trematodes, cestodes, nematodes and acanthocephalans to parasite assemblages. Although the species richness of helminth communities showed no consistent latitudinal variation, their taxonomic composition varied as a function of both host type and latitude. First, trematodes and acanthocephalans accounted for a higher proportion of species in helminth communities of fish, whereas nematodes achieved a higher proportion of the species in communities of bird and especially mammal hosts. Second, the proportion of trematodes in helminth communities of birds and mammals increased toward higher latitudes. Finally, the proportion of nematodes per community increased toward lower latitudes regardless of the type of host. We present tentative explanations for these patterns, and argue that new insights in parasite community ecology can be gained by searching for latitudinal gradients not only in parasite species richness, but also in the taxonomic composition of parasite assemblages.     

Pharmacokinetics of kanamycin in the bullfrog, Rana catesbeiana

1. Kanamycin disposition was studied in bullfrogs (Rana catesbeiana) following single doses IP. Both plasma t1/2 and Vd of the drug increased with increasing time after drug indicating redistribution and tight binding of kanamycin to deep tissue compartments. 2. Kanamycin was eliminated unchanged with a t1/2 plasma = 27 hr; perilymph = 89 hr; endolymph = 183 hr; aqueous humor = 54 hr; and CSF = 58 hr. 3. Kanamycin was absorbed by frogs from environmental water. 4. Environmental conditions must be carefully specified and monitored, as well as the physiological state of the animals when studying the effects of drugs on Amphibia.     

The emerging amphibian pathogen Batrachochytrium dendrobatidis globally infects introduced populations of the North American bullfrog, Rana catesbeiana

Batrachochytrium dendrobatidis is the chytridiomycete fungus which has been implicated in global amphibian declines and numerous species extinctions. Here, we show that introduced North American bullfrogs (Rana catesbeiana) consistently carry this emerging pathogenic fungus. We detected infections by this fungus on introduced bullfrogs from seven of eight countries using both PCR and microscopic techniques. Only native bullfrogs from eastern Canada and introduced bullfrogs from Japan showed no sign of infection. The bullfrog is the most commonly farmed amphibian, and escapes and subsequent establishment of feral populations regularly occur. These factors taken together with our study suggest that the global threat of B. dendrobatidis disease transmission posed by bullfrogs is significant.     

Peptides with antimicrobial activity from four different families isolated from the skins of the North American frogs Rana luteiventris, Rana berlandieri and Rana pipiens.

The skins of frogs of the genus Rana synthesize a complex array of antimicrobial peptides that may be grouped into eight families on the basis of structural similarity. A total of 24 peptides with differential growth-inhibitory activity towards the Gram-positive bacterium Staphylococcus aureus, the Gram-negative bacterium Escherichia coli and the yeast Candida albicans were isolated from extracts of the skins of three closely related North American frogs, Rana luteiventris (spotted frog), Rana berlandieri (Rio Grande leopard frog) and Rana pipiens (Northern leopard frog). Structural characterization of the antimicrobial peptides demonstrated that they belonged to four of the known families: the brevinin-1 family, first identified in skin of the Asian frog Rana porosa brevipoda; the esculentin-2 family, first identified in the European frog Rana esculenta; the ranatuerin-2 family, first identified in the North American bullfrog Rana catesbeiana; and the temporin family, first identified in the European frog Rana temporaria. Peptides belonging to the brevinin-2, ranalexin, esculentin-1 and ranatuerin-1 families were not identified in the extracts. Despite the close phylogenetic relationship between the various species of Ranid frogs, the distribution and amino-acid sequences of the antimicrobial peptides produced by each species are highly variable and species-specific, suggesting that they may be valuable in taxonomic classification and molecular phylogenetic analysis.     

Cytopathological observations and epizootiology of frog erythrocytic virus in bullfrogs (Rana catesbeiana).

A 5 yr survey (1985 to 1989) revealed that the prevalence of frog erythrocytic virus (FEV) was significantly higher in juveniles than in adult bullfrogs (Rana catesbeiana) in Algonquin Park, Ontario (Canada). The prevalence of infection in juveniles declined during May and June and increased in early August, persisting until late September. Tadpoles were not infected naturally with FEV, but were susceptible to experimental infection. Experimental data indicate that FEV is neither waterborne, nor transmitted by the oral route, nor by the leech Desserobdella picta. The virus may be transmitted mechanically by the mosquito Culex territans, or the midge Forcipomyia (Lasiohelea) fairfaxensis. Nine percent recapture of marked uninfected frogs versus 4% recapture of infected animals suggests that FEV contributes to mortality of juvenile bullfrogs. Infected erythrocytes were transformed from ellipsoidal to spheroidal cells, some of which contained flattened elongate, trapezoidal inclusions. There was no evidence of denatured hemoglobin or significant change in the hemoglobin content of infected cells. Anemia recorded in heavily infected animals was not attributable to an increased osmotic fragility of infected erythrocytes.     

Forelimb regeneration in the postmetamorphic bullfrog: stimulation by dimethyl sulfoxide and retinoic acid

To examine the effects of retinoic acid and dimethyl sulfoxide on regenerative ability of anuran amphibians, the left forelimbs of 60 postmetamorphic froglets of Rana catesbeiana (bullfrogs) were amputated through the distal zeugopodium. Fifteen of the froglets had their left forelimb stumps immersed in dimethyl sulfoxide (DMSO) for 3 minutes, once immediately after amputation and once on each of 5 subsequent days. Another 15 frogs had their left forelimb stumps immersed in a 0.01 M solution of retinoic acid dissolved in DMSO for the same period of time. The remaining 30 control froglets did not regenerate structures distal to the amputation surface, while all limbs in both treated groups produced regenerates by 120 days postamputation. Regenerates of limbs treated with both DMSO alone and DMSO combined with retinoic acid, although hypomorphic, were composed of multiple cartilage elements, which in many cases (46.7%) were organized as patterns partially resembling the skeletal arrangement of a normal forelimb. All of these regenerates exhibited bundles of striated muscle. In addition, nearly half (46.7%) of the regenerates in the DMSO + retinoic acid group possessed two separate regenerate outgrowths. The results demonstrate that young bullfrogs (Rana catesbeiana) possess a latent epimorphic regenerative capability, which can be stimulated by topical application to the wound surface of DMSO alone or DMSO combined with retinoic acid.     

Intestinal helminths of capybaras, Hydrochoerus hydrochaeris, from Venezuela

Quantitative parameters of intestinal helminth species and their potential relations to host characteristics in a population of capybaras (Hydrochoerus hydrochaeris) from Venezuela are reported for the first time. The intestines of 40 capybaras were collected during the 1992-annual harvest at Hato El Cedral. Six helminth species were found: 2 cestodes (Monoecocestus macrobursatum, M. hagmanni), 2 nematodes (Viannella hydrochoeri, Protozoophaga obesa), and 2 trematodes (Hippocrepis hippocrepis, Taxorchis schistocotyle). This is the first report for M. macrobursatum in Venezuela. Helminth abundance did not differ between sexes or age classes. Although patterns of distribution for all helminth species were overdispersed, the high prevalence found for all species (over 70%) and the high abundance observed for nematodes made it difficult to assess the effect that these helminths may produce on capybaras. Nevertheless, the negative associations found between the body condition of capybaras and helminth intensity for M. macrobursatum and V. hydrochoeri, might be pointing out potential host population regulatory role for these parasites which require further research.     

The Ability of Three Species of Tadpoles to Differentiate among Potential Fish Predators

The ability of prey to respond to novel predator cues may depend on the generality or specificity of the response to predator cues. We used laboratory behavioral experiments to examine the ability of tadpoles of three species of anurans (American toad, Bufo americanus ; bullfrog, Rana catesbeiana ; and green frog, R. clamitans ) to respond to the presence of two native potential predators (bluegill, Lepomis macrochirus ; and largemouth bass, Micropterus salmoides ) and one non-native potential predator (goldfish, Carassius auratus ). We also examined the effect of tadpole size on the behavioral responses of American toads and green frogs to predator cues. All three species of tadpoles responded to the presence of predator cues, although the specific responses varied among species. American toads and green frogs reduced activity in the presence of at least some fish cues, but bullfrog tadpoles did not change their activity. Bullfrogs decreased use of vegetation in the presence of some predator cues, whereas American toads and green frogs did not. American toads only responded to the presence of bluegill cues but not the other fish predator cues, whereas bullfrogs and green frogs responded more generally to the fish predators. In both American toads and green frogs, tadpole size affected behavior. For American toads, activity increased, as did the use of the vegetated side of the aquarium, in larger tadpoles. Not only did size affect American toad behavior, but it also influenced the responses of the tadpoles to predator cues. For green frogs, activity decreased in larger tadpoles. Our results suggest that behavioral responses of tadpoles to predator cues can be influenced by both the identity of the predator and the prey, as well as the size of the potential prey.