Effects of Predation by Invasive Western Mosquitofish(Gambusia affinis) on Survival of Eggs,Embryos and Tadpoles of Pelophylax nigromaculatus and Duttaphrynus melanostictus in South China

Alien species are one of the most serious threats to the decline and extinction of native amphibian populations. In this study, we examined the predation of invasive Western Mosquitofish Gambusia affinis on the eggs, embryos, and tadpoles of Duttaphrynus melanostictus and Pelophylax nigromaculatus in south China. Our results suggested that the survival of eggs and embryos remaining in the egg capsules of P. nigromaculatus and D. melanostictus was significantly higher than those removed from the egg capsule at 12-h intervals within 72 h in the presence of G. affinis. The survival of P. nigromaculatus eggs and embryos without egg capsules was significantly lower than those of D. melanostictus without egg capsules. The survival of P. nigromaculatus eggs and embryos with egg capsules was significantly higher than those of D. melanostictus with egg capsules from 24 h to 72 h except for 12 h. The survival of D. melanostictus tadpoles was significantly higher than that of P. nigromaculatus tadpoles in the presence of G. affinis. The survival of Gosner stage 26 tadpoles of P. nigromaculatus was significantly higher than that of Gosner stage 30 tadpoles from 12 h to 60 h, but there were no significant differences at 72 h. In contrast, the survival of Gosner stage 26 tadpoles of D. melanostictus was significantly lower than that of Gosner stage 30 tadpoles within 72 h, recording every 12 h. The increasing temperature caused a significant increase in predation by G. affinis on P. nigromaculatus eggs and embryos. The outer jelly capsule surrounding anurans eggs might serve as a mechanical defense against predation by G. affinis due to its large diameter, relatively stationary state and unpalatability. The differences in the vulnerability of P. nigromaculatus and D. melanostictus embryos and tadpoles to G. affinis probably due to differences in the unpalatability, black skin and activity. Based on the magnitude of predation by G. affinis on the eggs, embryos and tadpoles of these two species and the combined impact of temperature, we might speculate that invasive G. affinis and global warming would have more detrimental impacts on population viability of P. nigromaculatus than D. melanostictus in China.     

Distribution and reproductive strategies of Gyrinicola batrachiensis (Oxyuroidea: Pharyngodonidae) in larvae of eight species of amphibians from Nebraska

In total, 462 tadpoles and salamander larvae of 8 species were examined for the presence of Gyrinicola batrachiensis from 5 locations in Nebraska. Infection by G. batrachiensis occurred in tadpoles of Rana blairi , Rana catesbeiana, Rana pipiens, and Bufo woodhousii. Tadpoles of Hyla chrysoscelis , Spea bombifrons, and Pseudacris maculata and larvae of Ambystoma mavortium were not infected with G. batrachiensis. Population structure, defined as prevalence, mean abundance, and mean intensity of G. batrachiensis, varied among tadpoles of different amphibian species and was determined by collection locality, developmental period of tadpole hosts, amphibian species co-occurrence, and different reproductive strategies of G. batrachiensis , or a combination. Gyrinicola batrachiensis observed in all ranid tadpoles and B. woodhousii tadpoles from where bufonids were the only anuran species present, confirmed to the didelphic haplodiploidy and monodelphic parthenogenetic reproductive strategies, respectively. However, tadpoles of B. woodhousii that co-occurred with tadpoles of R. pipiens at Cedar Creek were inconsistent with these predictions and contained both male and didelphic female nematodes, but at a low mean intensity (1.61 ± 0.70). Didelphic female nematodes from B. woodhousii tadpoles at Cedar Creek only produced thick-shelled eggs, whereas nematodes in R. pipiens tadpoles had a high mean intensity (14.88 ± 23.83) from this location and contained both thick-shelled and thin-shelled eggs in their respective uteri. More importantly, adult female nematodes from tadpoles of R. pipiens and B. woodhousii from Cedar Creek were morphologically more similar to each other than to female nematodes recovered from tadpoles of other anuran species, other locations, or both. These data suggest that when strains of G. batrachiensis are shared by tadpoles of different amphibian species that differ in developmental period, the nematodes have an intermediate reproductive strategy in amphibian species, with tadpoles having short development.     

Extra-intestinal localization of Goussia sp. (Apicomplexa) oocysts in Rana dalmatina (Anura: Ranidae), and the fate of infection after metamorphosis

Although coccidia of the genus Goussia are common parasites of fish, only 2 species have been described in amphibians: G. hyperolisi from common reed frogs Hyperolius viridiflavus from Kenya and G. neglecta from unspecified European water frogs of the genus Rana from Germany. The genus Goussia is characterized by an oocyst, with a fine oocyst wall, containing 4 dizoic sporocysts that are composed of 2 valves joined by a longitudinal suture and lacking a Stieda body (typical for the genus Eimeria). To date, infections in amphibians were generally considered to be specific to the intestine of aquatic larval stages (tadpoles) of anurans. Herein, we report on: (1) the presence of oocysts of Goussia sp. in an extra-intestinal location (liver) of tadpoles of the agile frog R. dalmatina and (2) the presence of oocysts in the liver of both juvenile and subadult R. dalmatina. These observations represent novel traits for Goussia infections in amphibians; they may explain the vertical transmission of Goussia in tadpoles.     

Population divergence in growth rate and antipredator defences in Rana arvalis

Growth and development rates often differ among populations of the same species, yet the factors maintaining this differentiation are not well understood. We investigated the antipredator defences and their efficiency in two moor frog Rana arvalis populations differing in growth and development rates by raising tadpoles in outdoor containers in the nonlethal presence and absence of three different predators (newt, fish, dragonfly larva), and by estimating tadpole survival in the presence of free-ranging predators in a laboratory experiment. Young tadpoles in both populations reduced activity in the presence of predators and increased hiding behaviour in the presence of newt and fish. Older tadpoles from the slow-growing Gotland population (G) had stronger hiding behaviour and lower activity in all treatments than tadpoles from the fast-growing Uppland population (U). However, both populations showed a plastic behavioural response in terms of reduced activity. The populations differed in induced morphological defences especially in response to fish. G tadpoles responded with relatively long and deep body, short tail and shallow tail muscle, whereas the responses in U tadpoles were often the opposite and closer to the responses induced by the other predators. U tadpoles metamorphosed earlier, but at a similar size to G tadpoles. There was no evidence that growth rate was affected by predator treatments, but tadpoles metamorphosed later and at larger size in the predator treatments. G tadpoles survived better in the presence of free-ranging predators than U tadpoles. These results suggest that in these two populations, low growth rate was linked with low activity and increased hiding, whereas high growth rate was linked with high activity and less hiding. The differences in behaviour may explain the difference in survival between the populations, but other mechanisms (i.e. differences in swimming speed) may also be involved. There appears to be considerable differentiation in antipredator responses between these two R. arvalis populations, as well as with respect to different predators.     

化肥磷酸二铵对花背蟾蜍蝌蚪发育的影响

目的为了探究化肥磷酸二铵对两栖类幼体发育的影响。方法以花背蟾蜍(Bufo raddei)蝌蚪为研究对象,用化肥磷酸二铵(DAP)进行染毒培育,并依据Gosner(1960)分期标准,将蝌蚪分为G0实验组和G20实验组,分别暴露于浓度为0.045g/L、0.135g/L、0.225g/L的DAP水溶液中,直至蝌蚪完成变态发育,并对孵化率、死亡率、畸形率以及发育时程等参数进行统计分析。结果随着化肥DAP浓度的升高,孵化率逐渐下降,但对照组与各侵染组之间均无显著性差异;侵染组蝌蚪死亡率均高于对照组,均呈极显著性差异(P0.01),且G20侵染组明显高于G0侵染组;蝌蚪畸形可分为四大类,侵染组蝌蚪畸形率均高于对照组,且均呈极显著性差异(P0.01),畸形率随着DAP浓度的升高而增加;完成变态发育所用时程,侵染组明显长于对照组。结论化肥磷酸二铵对蝌蚪发育具有显著影响及损伤。     

Effects of high gravity on amphibian development.

In order to clarify the possible effects of high gravity environments on eggs and developing embryos, Rana rugosa and Xenopus laevis fertilized eggs and early embryos were raised in 2 G, 5 G, 7 G and 10 G up to the hatched tadpole stage. The results showed that: (1) High gravity significantly retarded the development of eggs and embryos beginning treatment before the blastula stage and induced various abnormalities, including two heads and microcephally suggesting that high gravity is apt to disrupt the animal-vegital axis. On the other hand, embryos beginning treatment after the gastrula stage showed a striking increase in the number of normal-appearing feeding tadpoles. (2) Autopsy revealed that brains, notochords and muscles were reduced in development and differentiation for embryos and tadpoles developed in high gravity. (3) It seems likely that the system for hydrogen peroxide detoxification develops abnormally in high gravity-treated embryos and tadpoles, which probably results in oxidative stress, leading to considerable cell damage.     

Characterization of chemical defenses in ranid tadpoles against a fish predator

Tadpoles of some ranid species appear to possess chemical defenses against fish predators, but the chemicals have not been characterized. Here, we evaluated the vulnerability of three Japanese anuran tadpole species (Glandirana rugosa, Pelophylax nigromaculatus, and Hyla japonica) to a fish (Gnathopogon elongatus elongatus) and analyzed the defensive chemicals extracted from the unpalatable tadpoles. Additionally, we examined the defensive behavior of unpalatable tadpoles in response to fish chemical cues. The fish rejected both G. rugosa (83%) and P. nigromaculatus (48%), but not H. japonica (0%). Many of the rejected tadpoles survived (60–80%). Possible defensive chemicals were extracted by methanol from the skin of G. rugosa, but were not identifiable by gas chromatography–mass spectrometry because of small quantities. The chemicals have high polarity and non-volatility. When exposed to fish chemical cues, P. nigromaculatus decreased activity presumably as a defensive behavior, but G. rugosa did not. We demonstrated the presence of chemical defenses in at least two of these species and revealed that G. rugosa releases more effective or greater amounts of defense chemicals than P. nigromaculatus with respect to this fish predator. The increased efficacy of chemical defenses may correlate with decreasing defensive behavior.     

Populational Changes of the Enteric Protozoans Opalina spp. and Nyctotherus cordiformis During the Ontogeny of Anuran Tadpoles

ABSTRACT. We studied ontogenetic population changes of Opalina and Nyctotherus cordiformis in eight species of tadpoles from 10 sites in east-central Mississippi. Most tadpoles acquired Opalina early in development, while the acquisition of N. cordiformis was variable. Developmental stage, species and collection site explained significant amounts of the variation in Opalina density of tadpoles ( F = 11.6; df = 27, 235; P < 0.0001) and metamorphs ( F = 7.31; df = 24, 84; P < 0.0001). Relationships between Opalina density and host stage showed either (1) a gradual decrease or (2) a gradual increase throughout host ontogeny. Opalina densities declined during metamorphosis. Density variations of N. cordiformis were explained by host species of tadpoles ( F = 9.30; df = 7, 142; P < 0.0001) and by host species and stage of metamorphs ( F = 5.85; df = 8, 62; P < 0.0001).
The length of larval period, habitat duration and generation time of the protozoans are suggested as major modifiers of the protozoan densities. Hosts with long larval periods show a decreasing population density and hosts with short developmental periods show a pattern of increasing density. Neither pattern was detected in tadpoles from temporary sites. Metamorphic declines in protozoan density, but not necessarily the loss of protozoans, reflect metamorphic alterations of the gut common to all hosts.     

New contact zone and assemblages among water frogs of Southern France

Water frogs are involved in several hybridogenetic complexes. However, investigations in Southern and Western France produced evidence of new hybridization events between R. lessonae and R. perezi (some new hybrids tadpoles were discovered), and the existence of new assemblages [R–G, L–P, R–P, P–R–G: (R,  ridibunda; G,  grafi; L, lessonae; P,  perezi )]. In respect to hybridogenesis, these assemblages offer opportunities for both primary and secondary hybridizations. Such peculiarities may be explained either by introductions or by relic populations and may influence water frog evolution.     

Metabolic compensation and behavioral thermoregulation of subtropical rhacophorid (Polypedates megacephalus) tadpoles in container habitats

We examined the relative importance of behavioral thermoregulation and metabolic compensation used by a subtropical rhacophorid (Polypedates megacephalus) tadpoles living in man-made container habitats to cope with thermal stress. We collected foam nests of P. megacephalus from man-made container habitats, and hatchlings were raised in 150 or 15 L of water (LWB and SWB containers, respectively). Water and air temperatures of containers were monitored using a datalogger. Tadpoles from both types of containers were acclimated at 22 and 32 degrees C for 10 d before measuring oxygen consumption (V(O2)) in a closed-system at 22, 27, and 32 degrees C. Thermal selection of tadpoles from two containers was determined using an aquatic thermal gradient. We observed daily stratification of temperature in the water column of LWB containers but not SWB containers. Tadpoles from LWB and SWB containers exhibited metabolic compensation so that tadpoles acclimated to 22 degrees C had significantly higher V(O2) than those acclimated to 32 degrees C. This was probably related to the variation of environmental temperature experienced by the tadpoles. Tadpoles of LWB and SWB containers selected similar water temperatures with low coefficient of variation, suggesting they are good thermal selectors. Results of this study suggest that P. megacephalus tadpoles use both behavioral thermoregulation and metabolic compensation to cope with the environmental temperature fluctuation, and this is, in part, due to the heterogeneity in the thermal regimes of breeding habitats. Even though metabolic compensation of tadpoles incurs a cost, P. megacephalus tadpoles that experience no daily thermal gradient in the man-made water bodies and/or seasonal variations in temperature over tadpole period evolve metabolic compensation to maintain physiological homeostasis under different thermal regimes.     

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.     

Temperature influence in developing tadpoles. II. Influence of temperature on isozymes and energies of activation of partially purified glucose-6-phosphate dehydrogenase

Isozymic banding patterns and energies of activation were studied in partially puriled G6PDH preparations from spring and fall Rana pipiens eggs and tadpoles raised at 10 °C and 20 °C.G6PDH preparations from spring tadpoles raised at 20 °C demonstrated new isozyme patterns. However, in the spring 10 °C and fall 10 °C and 20 °C raised tadpoles G6PDH preparations conserved isozymes already present.The energies of activation of G6PDH determined from the maximum velocity constants in the spring groups are lower than in the corresponding fall groups.     

Glucocorticosteroid hormone treatment of larval treefrogs increases infection by Alaria sp. trematode cercariae

In many amphibian species, an apparent increase has occurred in the prevalence of limb deformities caused by parasitic trematodes. We are interested in the role of environmental stressors in increasing these infections in amphibians. One mechanism by which environmental stressors could act to increase disease prevalence is to increase circulating levels of glucocorticosteroid hormones, which are released in response to stressors and can be immunosuppressive. In the present study, we treated gray treefroZg tadpoles (Hyla versicolor) with exogenous corticosterone, which is the main glucocorticosteroid "stress" hormone in amphibians. We then exposed treated tadpoles to Alaria sp. cercariae and scored the number of mesocercariae that successfully infected the tadpoles. In addition, we assayed one function of the immune response by counting the number of circulating eosinophilic granulocytes, which are thought to be important in immune responses to macroparasites. Tadpoles treated with exogenous corticosterone developed higher parasite loads than control tadpoles did, and they had lower numbers of circulating eosinophilic granulocytes. These results provide evidence of glucocorticosteroid-mediated immunosuppression in tadpoles that may help to explain apparent increases in the numbers of trematode-induced deformities in amphibian populations during recent decades.     

Androgen directs sexual differentiation of laryngeal innervation in developing Xenopus laevis

In adult Xenopus laevis, innervation of the vocal organ is more robust in males than in females. This sex difference originates during tadpole development; at stage 56, when the gonads first differentiate, the number of axons entering the larynx is the same in the sexes, but by stage 62, innervation is greater in males. To determine if androgen secretion establishes sex differences in axon number, we treated tadpoles with antiandrogen or androgen beginning at stage 48 or 54 and counted laryngeal nerve axons at stage 62 using electron microscopy. When male tadpoles were treated with the antiandrogen hydroxyflutamide, axon numbers were reduced to female-typical values; axon numbers in females were unaffected by antiandrogen treatment. When female tadpoles were treated with the androgen DHT (dihydrotestosterone), axon numbers were increased to male-like values. These findings suggest that endogenous androgen secretion during late tadpole stages in males is required for the sexual differentiation of laryngeal innervation observed from stage 62 on. Because androgen treatment and laryngeal innervation affect myogenesis in postmetamorphic frogs, numbers of laryngeal dilator muscle fibers were determined for hormonally manipulated tadpoles. At stage 62, vehicle-treated males had more laryngeal axons than females; laryngeal muscle fiber numbers did not, however, differ in the sexes. Both male and female tadpoles, treated from stage 54 with DHT, had more muscle fibers at stage 62 than vehicle-treated controls. Thus, while endogenous androgen secretion during late tadpole stages is subthreshold for the establishment of masculinized muscle fiber numbers, laryngeal myogenesis is androgen sensitive at this time and can be increased by suprathreshold provision of exogenous DHT. A subgroup of tadpoles, DHT treated from stage 54 to 62, was allowed to survive, untreated, until postmetamorphic stage 2 (PM2: 5 months after metamorphosis is complete). Androgen treatment between tadpole stages 54 and 62 does not prevent the ontogenetic decrease in axon numbers characteristic of laryngeal development. In addition, the elevation in stage 62 axon numbers produced by DHT-treatment at late tadpole stages was not associated with elevated numbers of laryngeal muscle fibers at PM2. Juvenile males normally maintain elevated axon numbers (relative to final adult values) through PM2 and the presence of these additional axons may result from-rather than contribute directly to—laryngeal muscle fiber addition. 1994 John Wiley & Sons, Inc.     

Expanded geographical range, new host accounts, and observations of the nematode Gyrinicola batrachiensis (Oxyuroidea: Pharyngodonidae) in tadpoles

We describe the first account of the nematode Gyrinicola batrachiensis in the gastrointestinal tracts of tadpoles of Bufo terrestris, Gastrophryne carolinensis, Hyla femoralis, H. squirella, Rana heckscheri, and R. utricularia. Infection by G. batrachiensis was also noted in tadpoles of R. catesbeiana and R. clamitans, species previously described as harboring these nematodes. These observations represent a major expansion of the known geographical range of G. batrachiensis to include the southeastern United States. In our study, female G. batrachiensis nematodes in ranid host species were didelphic, but females in B. terrestris tadpoles were monodelphic. Furthermore, only female nematodes were found in the latter host species. Such variations in nematode uterine morphology and sex ratios among these tadpole host species are in accordance with the apomictic and haplodiploid reproductive strategies previously reported for G. batrachiensis. These observations also support a generalized pattern relating G. batrachiensis reproductive modes and the life cycles of their tadpole hosts.     

黑斑侧褶蛙蝌蚪断尾后的补偿生长和发育研究

动物在经历不利的生长条件或环境后往往出现补偿生长。研究了黑斑侧褶蛙(Pelophylax nigromaculatus)蝌蚪经历来自食蚊鱼(Gambusia affinis)捕食造成断尾损伤后的补偿生长模式、断尾损失对蝌蚪游泳能力以及变态时间与大小的影响。结果表明,经历捕食压力后,全尾组和1/3断尾组蝌蚪的体长显著大于1/2断尾组蝌蚪的体长,全尾组和1/3断尾组之间的蝌蚪体长差异不显著;第19天时,1/2断尾组蝌蚪经过补偿生长后体长显著大于全尾组蝌蚪,1/3断尾组与全尾组和1/2断尾组之间的蝌蚪体长差异均不显著;三个实验组之间蝌蚪尾长差异不显著;全尾组蝌蚪的疾游速显著大于1/2断尾组蝌蚪的疾游速,1/3断尾组蝌蚪的疾游速与全尾组和1/2断尾组之间差异不显著,表明严重断尾对蝌蚪疾游速产生了消极影响。三个实验组蝌蚪的变态时间和变态前后形态差异均不显著。黑斑侧褶蛙蝌蚪能够在变态前调整生长轨迹补偿早期捕食风险造成的生长损失,断尾损失并不影响变态时间与大小,断尾超过一半的蝌蚪经过补偿生长后仍要付出一定的运动代价。     

Aldosterone upregulates purinergic responses in larval amphibian skin epithelium

Bullfrog tadpoles respond to apical application of 100 microM amiloride, acetylcholine (ACh) or ATP with a sharp transient inward (apical to basolateral) cation current. In adult skin, amiloride blockable transepithelial Na+ transport is upregulated by the hormone aldosterone. Tadpoles were treated in vivo with aldosterone and changes in short circuit current (Isc) in response to apical application of ATP were determined. Bullfrog tadpoles were exposed to aldosterone (10(-6) M) for periods ranging from 3 h to 60 h. Skins from 60-h aldosterone-treated animals showed a two- to three-fold increase in apical ATP-activated short circuit current when compared to animals treated with vehicle alone. Sodium replacement with a large, nonpermeable cation resulted in no measurable increase in Isc after exposure to ligand, consistent with ATP activation of an inward cation current and not chloride efflux. Activation/desensitization time courses and treatment with blockers revealed no measurable differences between aldosterone-treated and non-treated skins. Activation by amiloride and ACh gave essentially identical results. Studies with RT/PCR showed significant increases over controls of levels of mRNA associated with P2X channels. Given these data, our working hypothesis is that all three ligands activate the same process that exhibits both purinergic and cholinergic characteristics. These data are consistent with aldosterone upregulation of ATP gated channels expressed in the apical membrane of larval frog skin.     

Field Response of Tadpoles to Conspecific and Heterospecific Alarm

Many organisms use chemical cues from a variety of sources to mediate predator avoidance. Response to heterospecific alarm cues has been demonstrated for tadpoles within but not among taxa and alarm response behavior has seldom been examined under field conditions. This study examined the response of three sympatric amphibian larvae and predaceous larval Dytiscus sp. (diving beetle) to damage-release signals in natural ponds by using capture rates from treated funnel traps as an index of larval behavior. Hyla regilla (Pacific tree frog) tadpoles avoided traps treated with either crushed conspecifics or with Rana aurora (red-legged frog) tadpoles but the larger ranids and Ambystoma macrodactylum (long-toed salamander) did not respond to either treatment. H. regilla tadpoles were likely susceptible to any potential predators of ranid tadpoles in these ponds and this result is consistent with the hypothesis that a response to heterospecific alarm occurs in sympatric prey with shared predators.     

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.     

The differential influence of protein kinase inhibitors on retinal arbor morphology and eye-specific stripes in the frog retinotectal system

We investigated retinal axon morphology and eye-specific afferent termination zones in the optic tectum of three-eyed tadpoles that were chronically treated with protein kinase inhibitors. The kinase inhibitors sphingosine, H-7, and phorbol ester, which down-regulates protein kinase C with chronic exposure, were applied to the tecta in a slow release plastic, Elvax. In vivo protein phosphorylation assays in drug-treated tadpoles indicated that the treatments decreased 32P incorporation into some protein bands by as much as 60%. Although the drugs did not cause a desegregation of the eye-specific stripes, treated retinal axon arbors covered about half the area covered by untreated arbors or arbors treated with inactive analogs of the drugs. We conclude that eye-specific segregation can be maintained under conditions that markedly alter retinal ganglion cell axon arbor size and that significantly perturb protein phosphorylation. Furthermore, we conclude that the protein kinase(s) that we blocked with these treatments is involved in the growth of axon arbors.