Tadpole Growth Research Articles

Cohort patterns in Rana tadpole metamorphosis

AbstractThis study explores the cohort patterns in a size versus time plot of individual metamorphs of Rana tadpoles. This meta‐study was based on a number of studies where any of three species of Rana tadpoles were raised in tanks under various treatments (variation in source pond, tadpole density, drying regime, hatching date, food level and competitors). A frequent pattern was found where there was an initial decrease in metamorph size as more slowly developing tadpoles metamorphosed. This negative trend sometimes changed to positive as later metamorphs appeared. The later metamorphs were often as large, or even larger, than those first to metamorphose, giving the plot a U‐shaped pattern. Also, a number of field studies of two of the Rana species’ were analysed for the same aspect. The U‐shaped pattern was much rarer but it did occur. A positive linear slope, rather than a negative one, was also found occasionally. A large number of factors affect the growth and development rates in tadpoles, adaptively and non‐adaptively, and thus determine their size at the time of metamorphosis. How these can contribute to the found metamorphosis patterns is discussed.

Growth strategies of tadpoles along the pond permanency gradient

The preference for particular features of water bodies for reproduction is one of the most important aspects of anuran ecology, affecting key aspects of both tadpole and adult life. The use by species of different habitats along the pond permanency gradient has been already studied, noting conflicting selective pressures from predation and desiccation risk. Here, we aim to discover physiological patterns related with this gradient. As a study system, we used the full anuran community of the NE Iberian Peninsula. We quantified growth rate, consumption rate, food assimilation and the proportion of energy allocated to growth, as well as gut length, for all species. Food consumption rate and growth allocation were the variables that defined tadpole growth, while food assimilation abilities and gut length seem to have a secondary or cryptic role in growth. More interestingly however, our data suggests a labile continuum of consumption-based versus allocation/assimilation-based growth strategies differentiating species. Differences among species follow predictions of adaptation to the pond permanency gradient selective pressures. Species from ephemeral ponds are more prone to use consumption-related growth tactics while species inhabiting ponds with longer hydroperiods are more efficient retaining and allocating energy into growth, although results seem partly shaped by strong interspecific competition. Physiological differences in growth and the usage of the assimilated energy could be an additional factor to understand how tadpoles adapt to the features of ponds they inhabit, as well as how they compete and coexist.

Effect of corticosterone on larval growth, antipredator behaviour and metamorphosis of Hylarana indica

Corticosterone (CORT), a principal glucocorticoid in amphibians, is known to regulate diverse physiological processes including growth and metamorphosis of anuran tadpoles. Environmental stressors activate the neuroendocrine stress axis (hypothalamus–pituitary–interrenal axis, HPI) leading to an acute increase in CORT, which in turn, helps in coping with particular stress. However, chronic increase in CORT can negatively affect other physiological processes such as growth and metamorphosis. Herein, we studied the effect of exogenous CORT on larval growth, antipredator behaviour and metamorphic traits of Hylarana indica. Embryonic exposure to 5 or 20μg/L CORT did not affect their development, hatching duration as well as larval growth and metamorphosis. Exposure of tadpoles to 10 or 20μg/L CORT throughout larval development caused slower growth and development leading to increased body mass at stage 37. However, body and tail morphology of tadpoles was not affected. Interestingly, larval exposure to 5, 10 or 20μg/L CORT enhanced their antipredator response against kairomones in a concentration-dependent manner. Further, larval exposure to increasing concentrations of CORT resulted in the emergence of heavier froglets at 10 and 20μg/L while, delaying metamorphosis at all concentrations. Interestingly, the heavier froglets had shorter hindlimbs and consequently shorter jump distances. Tadpoles exposed to 20μg/L CORT during early, mid or late larval stages grew and developed slowly but tadpole morphology was not altered. Interestingly, exposure during early or mid-larval stages resulted in an enhanced antipredator response. These individuals metamorphosed later but at higher body mass while SVL was unaffected.

Additive effects of mean temperature, temperature variability, and chlorothalonil to red-eyed treefrog (Agalychnis callidryas) larvae.

Amphibian populations are declining globally, and multiple anthropogenic stressors, including contamination by pesticides and shifting climates, are driving these declines. Climate change may increase average temperatures or increase temperature variability, either of which may affect the susceptibility of nontarget organisms to contaminants. Eight-day ecotoxicological assays were conducted with red-eyed treefrog (Agalychnis callidryas) larvae to test for additive and interactive effects of exposure to the fungicide chlorothalonil, average temperature, and temperature variability on tadpole growth and survival. Egg masses were collected from seasonal ponds at La Selva Biological Station in Costa Rica, and tadpoles were exposed to a series of chlorothalonil concentrations across a range of ecologically relevant mean temperatures (23.4-27.3 °C) and daily temperature fluctuations (1.1-9.9 °C). Survival was measured each day, and tadpole growth was measured at the end of each trial. Concentrations of chlorothalonil ≥60 µg/L reduced survival, although survival was not affected by mean temperature or daily temperature range, and there were no synergistic interactions between chlorothalonil and temperature regime on survival. Chlorothalonil suppressed tadpole growth at relatively low concentrations (∼15 µg/L). There were impacts of both average temperature and daily temperature range on tadpole growth, although there were no synergistic interactions between temperature regimes and chlorothalonil. The results should inform efforts to manage ecosystems impacted by multiple large-scale anthropogenic stressors as well as methods for the design of ecologically appropriate toxicology trials. Environ Toxicol Chem 2016;35:2998-3004. © 2016 SETAC.

Temperature-Dependence of Glyphosate-Based Herbicide's Effects on Egg and Tadpole Growth of Common Toads

Glyphosate-based herbicide formulations are broadly used in agriculture, silviculture, horticulture as well as in private gardens all over the world, thus posing the risk of potential contamination of nearby aquatic bodies inhabited by amphibians. Concurrently, climate change can be expected to alter the temperature of amphibian breeding sites. However, while either glyphosate-based herbicides or temperature have been shown to separately affect the development of amphibians, very little is known on possible interactive effects. We studied the impact of herbicide concentrations and temperature on growth and development of eggs and tadpoles of the Common toad (Bufo bufo L.). We hypothesized that (i) eggs would be better protected against herbicides than tadpoles because of their jelly coating, (ii) that higher temperatures would reduce potential herbicide effects because of an accelerated growth and a lower sensitivity of larger specimens. We conducted one experiment starting with eggs (Gosner stage, GS 9) and another experiment starting with tadpoles (GS 21-24) using a full factorial design with 5 concentrations of the herbicide formulation Roundup® LB Plus (0 mg acid equivalent L-1, 0.5 mg a.e. L-1, 1.0 mg a.e. L-1 or 1.5 mg a.e. L-1 and a pulse treatment with 3-4 times addition of 0.5 a.e. mg L-1 over the course of several weeks) and two temperature levels (15°C and 20°C). Contrary to our expectation, our results showed that toad eggs are more sensitive to herbicides than tadpoles leading to an averaged 31% increase in total length, tail length and body length compared to the herbicide-free control. Tadpole morphology, development or mortality was not influenced by herbicides. Higher temperature accelerated growth of both eggs and tadpoles. This is among the first study showing interactive effects between herbicides and temperature especially for egg development resulting in more pronounced herbicide effects at lower temperatures than at higher temperatures.

Dietary antioxidants enhance immunocompetence in larval amphibians

Dietary antioxidants have been shown to confer a variety of benefits through their ability to counter oxidative stress, including increased immunocompetence and reduced susceptibility to both infectious and non-infectious diseases. However, little is known about the effects of dietary antioxidants on immune function in larval amphibians, a group experiencing worldwide declines driven by factors that likely involve altered immunocompetence. We investigated the effects of dietary antioxidants (quercetin, vitamin E, and β-carotene) on two components of the immune system, as well as development and growth. Lithobates pipiens tadpoles fed diets with supplemental β-carotene or vitamin E exhibited an enhanced swelling response as measured with a phytohemagglutinin assay (PHA), but there was no induced antibody response. Effects were often dose-dependent, with higher antioxidant levels generally conferring stronger swelling that possibly corresponds to the innate immune response. Our results indicate that the antioxidant content of the larval amphibian diets not only had a detectable effect on their immune response capability, but also promoted tadpole growth (mass gain), although developmental stage was not affected. Given that many environmental perturbations may cause oxidative stress or reduce immunocompetence, it is critical to understand how nutrition may counter these effects.

Biotic Resistance to an Alien Amphibian: Larval Competition between Japanese Frogs and Invasive Cane Toads.

Understanding negative effects of native species on introduced taxa may suggest novel ways to control the invasive species by enhancing such effects. Previous studies have reported that the larvae of invasive cane toads (Rhinella marina) are suppressed by competition with the larvae of native anurans in Australia, but not in North America. We conducted laboratory trials to measure the effect of exposure to the larvae of Japanese frogs (Microhyla ornata, Fejervarya sakishimensis, Rhacophorus owstoni) on rates of survival, growth and development of cane toad tadpoles in Ishigaki Island, in southern Japan. Survival rates were not affected by native species, but competition with Dicroglossids and Rhacophorids (but not Microhylids) strongly reduced rates of growth and development in the tadpoles of cane toads. Dicroglossid tadpoles also reduced the body condition to toad tadpoles in addition to effects on SVL and mass. Encouraging populations of native frogs in toad-invaded areas of Japan thus may help to reduce the numbers of invasive cane toads.

The Effects of Crayfish Predation and Vegetation Cover on Tadpole Growth, Survival, and Nonlethal Injury

Abstract Predator–prey interactions play an important role in structuring larval anuran communities in ephemeral wetlands. The type of interaction often depends on the predator species and the complexity of the aquatic habitat. We experimentally evaluated the effects of Panhandle Crayfish (Procambarus evermanni) predation on larval Southern Leopard Frogs (Lithobates sphenocephalus) and larval Ornate Chorus Frogs (Pseudacris ornata). We performed separate experiments for each anuran species and used multiple vegetation treatments to examine whether vegetation could provide refugia from crayfish predation. Our results indicate that crayfish are effective predators of anuran larvae regardless of the amount of vegetation present. Encounters between tadpoles and crayfish often resulted in nonlethal tail injury for tadpoles, suggesting that crayfish predation is prominent in ephemeral wetland communities. Leopard frog tadpoles in predator treatments also grew larger than their counterparts in nonpredator treatm...

A reliable, non-invasive technique for measuring growth in tadpoles exposed to salt.

The use of chemical de-icers raises salt levels in roadside streams and ponds, which has adverse effects on tadpole development. Experiments on the effects of de-icers on tadpole development are often hampered by difficulties measuring body size without introducing handling stress that may skew results or cause unintended mortality. We have found a linear relationship between surface area and body mass in tadpoles that is unaffected by exposure to salt. Measuring surface area is therefore a suitable technique whose use should be encouraged when investigating the effects of salt exposure on tadpole growth and development.

Effects of different stocking density and diet on development of Bufo gargarizans tadpoles

The effects of four different stocking densities and five different diets on the growth of Bufo gargarizans tadpoles were studied to determine the optimum stocking density and diet. For stocking density experiment, the tadpoles were fed respectively at different density of 200, 500, 1 000 and 2 000 tadpoles per square meter. For diet experiment, the tadpoles were divided into five groups fed respectively with five different diets. The body weight, snout-vent length and tail length were measured every seven days, and mortality was recorded. The results showed that: the survival rates of tadpole before metamorphosis and after metamorphosis were from 68.7% to 96.3% and from 5.7% to 36.0%, respectively; the optimum stocking density is 1 000 tadpoles per square meter for the stocking density had no effect on the survival rate of tadpole before metamorphosis, and the tadpoles had the relative large body weight and survival numbers in 1000 tadpoles per square meter; the diet Ⅱ(37.9% crude protein and 5.7% crude fat), Ⅳ (25.1% crude protein and 4.0% crude fat), and Ⅴ (egg yolk) were all the optimum diets for the diet had no effect on the survival rate of tadpole before metamorphosis and the tadpoles fed with three kinds of diet above had relatively large body weight, and one of these three diets based on their availability and cost should be adopted during breeding period.

Canopy cover and anurans: nutrients are the most important predictor of growth and development

Bottom-up effects such as the availability of light and nutrients can have large impacts on primary producer quantity and quality, which is then translated into the growth and development of consumers. The use of “canopy cover” as a bottom-up predictive factor is a broad categorization, as canopy cover controls both the amount of light allowed into a pond and the nutrient load through leaf litter. To test how light and nutrients influence pond ecosystems, we manipulated inorganic nutrients and light in a 2 × 3 full-factorial, large-scale mesocosm experiment. Larval American Bullfrogs (Lithobates catesbeianus (Shaw, 1802)) were reared for 6 weeks at low densities and then assessed for development, growth, and survival at the end of the experiment. We also collected weekly samples of potential food resources (phytoplankton and periphyton) for the estimation of algal production and stoichiometric quality (carbon:nitrogen:phosphorus). Light had strong effects on food resource quality; however, resource quality did not significantly predict tadpole growth or development. Instead, nutrients seemed to be the most important factor as a stimulator of total algal primary production and some unknown pathway, which in turn affected tadpole development. Ours is the first study to investigate canopy cover using a comprehensive causal model, and our results suggest in regards to tadpole growth and development, canopy cover is important mainly as a source of nutrients to ponds.

Climate change will alter amphibian‐mediated nutrient pathways: evidence from Rana temporaria tadpoles in experimental ponds

Summary With global warming, mean temperatures and brownification of many waterbodies are predicted to increase. This may have unknown consequences on aquatic consumer life histories and nutrient content, consumer‐mediated nutrient recycling, and nutrient transport between water and land. Using a large‐scale experimental pond facility, we altered temperature (ambient/+4 °C) and brownification (clear/humic) in a 2 × 2 factorial design (n = 16 pond sections) to test two aspects of climate change on Rana temporaria tadpole life‐history traits and on tadpole‐mediated nutrient pathways. On day 16 after hatching, we examined tadpole‐mediated nutrient recycling by measuring tadpole nutrient excretion and egestion rates and tadpole body nutrient content. We estimated tadpole growth and development rates from hatching to emergence and measured emergent frog body size and body nutrient content. Brownification increased total pond water nutrient availability and total pond water nitrogen (N) : phosphorous (P) ratios. Warming positively affected tadpole growth and development rates, whereas browning increased tadpole growth rate only under ambient temperatures. Emergent frog body P content decreased with warming, but only in the clear treatments. But despite these variations in body nutrient content, body stoichiometry remained within a relatively narrow stoichiometric range for both emergent frogs (P content: 1.4–1.8%, N content: 11.4–11.8% and carbon [C] content: 46.9–51.3%) and tadpoles (P content: 1.1–1.2%, N content: 10.1–11.7% and C content: 48.0–50.5%). Warming increased tadpole body P content and browning had a positive effect on tadpole body N content and tadpole N excretion rates, probably mediated by the increased pond water total N availability. We conclude that warming and brownification will interact in changing aquatic consumer growth and body nutrient stoichiometry. In addition, warming has the potential to affect emergent frog body nutrient content and may thus affect nutrient transport from water to land. Last, by increasing pond water N availability, brownification appears to intensify consumer P limitation and thus amplify consumer‐meditated N recycling.

Differences in developmental strategies between long-settled and invasion-front populations of the cane toad in Australia.

Phenotypic plasticity can enhance a species' ability to persist in a new and stressful environment, so that reaction norms are expected to evolve as organisms encounter novel environments. Biological invasions provide a robust system to investigate such changes. We measured the rates of early growth and development in tadpoles of invasive cane toads (Rhinella marina) in Australia, from a range of locations and at different larval densities. Populations in long-colonized areas have had the opportunity to adapt to local conditions, whereas at the expanding range edge, the invader is likely to encounter challenges that are both novel and unpredictable. We thus expected invasion-vanguard populations to exhibit less phenotypic plasticity than range-core populations. Compared to clutches from long-colonized areas, clutches from the invasion front were indeed less plastic (i.e. rates of larval growth and development were less sensitive to density). In contrast, those rates were highly variable in clutches from the invasion front, even among siblings from the same clutch under standard conditions. Clutches with highly variable rates of growth and development under constant conditions had lower phenotypic plasticity, suggesting a trade-off between these two strategies. Although these results reveal a strong pattern, further investigation is needed to determine whether these different developmental strategies are adaptive (i.e. adaptive phenotypic plasticity vs. bet-hedging) or instead are driven by geographic variation in genetic quality or parental effects.

Conspecific visual cues: the relative importance of interference and exploitation competition among tadpoles of Rana kukunoris

Anuran larvae may use three types of cues (i.e. chemical, visual and tactile) in intraspecific competition, and subsequently mediate their growth and development; these cues are poorly understood and controversial. Here, we investigate the relative contribution of these cues in interference and exploitative mechanisms among Rana kukunoris tadpoles. In particular, we isolate the importance of visual interference signals against direct exploitation. Our results demonstrate that larvae reared alone and blinded to conspecifics had a longer larval period and grew larger than tadpoles reared alone but with conspecific visual cues or compared to tadpoles reared with conspecifics. Thus, we show evidence of visual interference competition, in which conspecific visual signals inhibit the growth of tadpoles and shorten larval development but do not reduce survival. We suggest tadpoles of R. kukunoris may use visual cues throughout ontogeny, delaying larval development to maximize individual fitness when environmental conditions (abundant food and space) allow for increased growth in the presence of few conspecifics.

Risk assessment based on indirect predation cues: revisiting fine-grained variation.

To adaptively express inducible defenses, prey must gauge risk based on indirect cues of predation. However, the information contained in indirect cues that enable prey to fine-tune their phenotypes to variation in risk is still unclear. In aquatic systems, research has focused on cue concentration as the key variable driving threat-sensitive responses to risk. However, while risk is measured as individuals killed per time, cue concentration may vary with either the number or biomass killed. Alternatively, fine-grained variation in cue, that is, frequency of cue pulses irrespective of concentration, may provide a more reliable signal of risk. Here, we present results from laboratory experiments that examine the relationship between red-eyed treefrog tadpole growth and total cue, cue per pulse, and cue pulse frequency. We also reanalyze an earlier study that examined the effect of fine-grained variation in predator cues on wood frog tadpole growth. Both studies show growth declines with increasing cue pulse frequency, even though individual pulses in high-frequency treatments contained very little cue. This result suggests that counter to earlier conclusions, tadpoles are using fine-grained variation in cue arising from the number of predation events to assess and respond to predation risk, as predicted by consumer-resource theory.

Effects of nutrient supplementation on host-pathogen dynamics of the amphibian chytrid fungus: a community approach.

Anthropogenic stressors may influence hosts and their pathogens directly or may alter host-pathogen dynamics indirectly through interactions with other species. For example, in aquatic ecosystems, eutrophication may be associated with increased or decreased disease risk. Conversely, pathogens can influence community structure and function and are increasingly recognised as important members of the ecological communities in which they exist.In outdoor mesocosms, we experimentally manipulated nutrients (nitrogen and phosphorus) and the presence of a fungal pathogen, Batrachochytrium dendrobatidis (Bd), and examined the effects on Bd abundance on larval amphibian hosts (Pseudacris regilla: Hylidae), amphibian traits and community dynamics. We predicted that resource supplementation would mitigate negative effects of Bd on tadpole growth and development and that indirect effects of treatments would propagate through the community.Nutrient additions caused changes in algal growth, which benefitted tadpoles through increased mass, development and survival. Bd-exposed tadpoles metamorphosed sooner than unexposed individuals, but their mass at metamorphosis was not affected by Bd exposure. We detected additive rather than interactive effects of nutrient supplementation and Bd in this experiment.Nutrient supplementation was not a significant predictor of infection load of larval amphibians. However, a structural equation model revealed that resource supplementation and exposure of amphibians to Bd altered the structure of the aquatic community. This is the first demonstration that sublethal effects of Bd on amphibians can alter aquatic community dynamics.

Deconstructing cartilage shape and size into contributions from embryogenesis, metamorphosis, and tadpole and frog growth.

Understanding skeletal diversification involves knowing not only how skeletal rudiments are shaped embryonically, but also how skeletal shape changes throughout life. The pharyngeal arch (PA) skeleton of metamorphosing amphibians persists largely as cartilage and undergoes two phases of development (embryogenesis and metamorphosis) and two phases of growth (larval and post-metamorphic). Though embryogenesis and metamorphosis produce species-specific features of PA cartilage shape, the extents to which shape and size change during growth and metamorphosis remain unaddressed. This study uses allometric equations and thin-plate spline, relative warp and elliptic Fourier analyses to describe shape and size trajectories for the ventral PA cartilages of the frog Xenopus laevis in tadpole and frog growth and metamorphosis. Cartilage sizes scale negatively with body size in both growth phases and cartilage shapes scale isometrically or close to it. This implies that most species-specific aspects of cartilage shape arise in embryogenesis and metamorphosis. Contributions from growth are limited to minor changes in lower jaw (LJ) curvature that produce relative gape narrowing and widening in tadpoles and frogs, respectively, and most cartilages becoming relatively thinner. Metamorphosis involves previously unreported decreases in cartilage size as well as changes in cartilage shape. The LJ becomes slightly longer, narrower and more curved, and the adult ceratohyal emerges from deep within the resorbing tadpole ceratohyal. This contrast in shape and size changes suggests a fundamental difference in the underlying cellular pathways. The observation that variation in PA cartilage shape decreases with tadpole growth supports the hypothesis that isometric growth is required for the metamorphic remodeling of PA cartilages. It also supports the existence of shape-regulating mechanisms that are specific to PA cartilages and that resist local adaptation and phenotypic plasticity.

Effects of resource distribution on the cost of predator avoidance behaviour in tadpoles

Tadpoles reduce activity and increase hiding in the presence of dragonfly larvae. Several studies demonstrate that tadpoles showing this behaviour have slower growth, however, other studies have found no effect or even positive growth in tadpoles exposed to predators. A recent study demonstrated that swimming is an energetically expensive activity for Bufo arabicus tadpoles. Therefore, if food resources are abundant close to refuges, reduced activity may be an advantage and could offset the cost of reduced foraging. I tested this hypothesis by growing B. arabicus tadpoles with food provided either near or away from shelters, in the presence or absence of caged dragonfly larvae. In the presence of dragonfly larvae, tadpoles provided with food close to shelters were significantly larger than those with food further away. Control tadpoles under both food treatments were intermediate in size, although not statistically different from the predator + near food tadpoles. The results indicate that access to resources is the main determinant of growth in B. arabicus tadpoles and that the energetic cost of swimming is a secondary factor.

The Acid Test: pH Tolerance of the Eggs and Larvae of the Invasive Cane Toad (Rhinella marina) in Southeastern Australia.

Invasive cane toads are colonizing southeastern Australia via a narrow coastal strip sandwiched between unsuitable areas (Pacific Ocean to the east, mountains to the west). Many of the available spawning sites exhibit abiotic conditions (e.g., temperature, salinity, and pH) more extreme than those encountered elsewhere in the toad's native or already invaded range. Will that challenge impede toad expansion? To answer that question, we measured pH in 35 ponds in northeastern New South Wales and 8 ponds in the Sydney region, in both areas where toads occur (and breed) and adjacent areas where toads are likely to invade, and conducted laboratory experiments to quantify effects of pH on the survival and development of toad eggs and larvae. Our field surveys revealed wide variation in pH (3.9-9.8) among natural water bodies. In the laboratory, the hatching success of eggs was increased at low pH (down to pH 4), whereas the survival, growth, and developmental rates of tadpoles were enhanced by higher pH levels. We found that pH influenced metamorph size and shape (relative head width, relative leg length) but not locomotor performance. The broad tolerance range of these early life-history stages suggests that pH conditions in ponds will not significantly slow the toad's expansion southward. Indeed, toads may benefit from transiently low pH conditions, and habitat where pH in wetlands is consistently low (such as coastal heath) may enhance rather than reduce toad reproductive success. A broad physiological tolerance during embryonic and larval life has contributed significantly to the cane toad's success as a widespread colonizer.

Low salinity increases survival, body weight and development in tadpoles of the Chinese edible frogHoplobatrachus rugulosus

Water salinity affects survival, growth and metamorphosis of anuran tadpoles. Hoplobatrachus rugulosus is considered not only as a freshwater amphibian but is also found in brackish wetlands. However, whether salinity change interferes with hatching, survival, body mass and development of H. rugulosus tadpoles is unknown. We found that salinity levels of <4‰ did not affect of survival or hatching of H. rugulosus eggs. At an early larval stage, tadpoles could tolerate up to 9‰ salinity for 96 h; however, body water content decreased when salinity was >5‰. After a 3-week experiment, body weights of tadpoles exposed to 2‰ and 4‰ salinities were higher but that of the 6‰ group was lower compared with the 0‰ group. More than 90% of tadpoles exposed to 2‰ and 4‰ salinity showed complete metamorphosis. Salinity levels <4‰ promoted survival of tadpoles better than 0‰, whereas none of tadpoles in the 6‰ group became juvenile frogs in 50 days. Time taken to reach metamorphosis was shorter for 2‰ and 4‰ (47.22 ± 0.28 and 47.26 ± 0.33 days, respectively) than for 0‰ (49.31 ± 0.35 days). Juvenile frogs in the 2‰ group had greater body weight than the control. It could be concluded that salinity of <4‰ increased survival and body weight of H. rugulosus tadpoles, and shortened the time taken to reach metamorphosis.