Frequent Anomalies in Larvae of Common and Moor Frogs in Moscow area and in the Suburbs of Moscow, Russia

Elena A. Severtsova, David R. Aguillon-Gutierrez, Alexsey S. Severtsov


We studied influence of anthropogenic pollution of spawning ponds on embryogenesis of the common and the moor frogs (Rana temporaria Linnaeus 1758, and Rana arvalis Nilsson 1852) from the natural population of the urban and the suburban territories of Moscow. Counts of larvae with anomalies in samples, fixed at different stages, indicated that in Moscow city and in the suburban territory the anomalies occur with approximately equal frequency. Spinal curvature, bradygenesis and skin damage were the most common anomalies. Edema (hydropsy), anomalies in gills, pigmentation disorders and complete malformation were also observed. In both species at all sites, most of the larvae had only one kind of anomaly, but some of them had two or more. Morphometric analysis of the spinal curvature showed that in abnormal tadpoles the angle between the body planes and tail plates was significantly less than in normal tadpoles, and is observed the underdevelopment of the tail. This anomaly is lethal because such embryos are not able to swim normally at later stages of development. Histological studies showed that the causes of spinal curvature in dorso-ventral direction is usually caused by somitogenesis defects or incomplete rotation of somites. Differences between species were observed in frequency of the less common anomalies. For Rana temporaria, edema, as well as a combination of complete malformation and skin defects were typical, while for Rana arvalis — a combination of edema and slow time of development was most common. No significant differences in frequency and nature of the anomalies were found between these species. However, inter-annual variability was remarkable. This suggests that frequency of occurrence of anomalies depends on the annual environmental situation.


Rana temporaria; Rana arvalis; anomalies; development; pollution

Full Text:



Abbasi S. A. and Soni R. (1984), «Teratogenic effects of chromium (VI) in environment as evidenced by the impact on larvae of amphibian Rana tigrina: Implications in the environmental management of chromium», Int. J. Environm. Studies, 23, 131 – 137.

Afonin B., Ho M., Gustin J. K., Meloty-Kapella C., and Domingo C. R. (2006), «Cell behaviour associated with somite segmentation and rotation in Xenopus laevis», Dev. Dynamics, 235, 3268 – 3279.

Allentoft M. E. and O’Brien J. (2010), «Global amphibian declines, loss of genetic diversity and fitness: a review», Diversity, 2, 47 – 71.

Allran J. W. and Karasov W. H. (2001), «Effects of atrazine on embryos, larvae and adults of anuran amphibians», Environ. Toxicol. Chem., 20, 769 – 775.

Ankley G. T., Tietge J. E., DeFoe D. L., Jensen K. M., Holcombe G. W., Durhan E. J., and Diamond S. A. (1998), «Effects of ultraviolet light and methoprene on survival and development of Rana pipiens», Environ. Toxicol. Chem., 17, 2530 – 2542.

Ankley G. T., Tietge J. E., Holcombre G. H., DeFoe D. L., Diamond S. A., Jensen K. M., and Degitz S. J. (2000), «Effects of laboratory ultraviolet radiation and natural sunlight on survival and development of Rana pipiens», Can. J. Zool., 78, 1092 – 1100.

Aronzon C. M., Sandoval M. T., Herkovits J., and Perez-Coll C. S. (2011), «Stage-dependent susceptibility to copper in Rhinella arenarum embryos and larvae», Environ. Toxicol. Chem., 30, 2771 – 2777.

Bacon J. P., Bacon D. W., Bacon R. L., and Bacon D. J. (2006), «Deformities in cane toad (Bufo marinus) populations in Bermuda: Part I. Frequencies and distribution of abnormalities», Appl. Herpetol., 3, 39 – 65.

Balmory A. (2010), «Mobile phone mast effects on common frog (Rana temporaria) tadpoles: the city turned into a laboratory», Electromag. Biol. Med., 29, 31 – 35.

Bancroft R. and Prahlad K. V. (1973), «Effect of ethylenebis [dithiocarbamic acid] disodium salt (nabam) and ethylenebis [dithiocarbamato] manganese (maneb) on Xenopus laevis development», Teratology, 7, 143 – 150.

Blaustein A. R. and Bancroft B. A. (2007), «Amphibian population declines: evolutionary considerations», BioScience, 57, 437 – 444.

Blaustein A. R. and Johnson P. T. J. (2003), «The complexity of deformed amphibians», Front. Ecol. Environ., 1, 87 – 94.

Blaustein A. R. and Wake D. B. (1995), «The puzzle of declining amphibian populations», Sci. American, 272, 52 – – 57.

Blaustein A. R., Kiesecker J. M., Chivers D. P., and Anthony R. G. (1997), «Ambient UV-B radiation causes deformities in amphibian embryos», PNAS, 94, 13735 – 13737.

Blaustein A. R., Wake D., and Sousa W. (1994), «Amphibian declines: judging stability, persistence, and susceptibility of populations to local and global extinctions», Conserv. Biol., 8, 60 – 71.

Bonin J., Ouellet M., Rodriguez J., Desgranges J., Gagne F., Sharbel T. F., and Lowcock L. A. (1997), «Measuring the health of frogs in agriculture habitats subjected to pesticides. Herpetological conservation», in: Green D. M., Amphibians in Decline: Canadian Studies of a Global Problem, pp. 141 – 148.

Bragg A. N. (1960), «Population fluctuations in the amphibian fauna of Cleveland County, Oklahoma, during the past twenty-five years», Southwest. Nat., 5, 165 – 169.

Bryer P. J., Elliott J. N., and Willingham E. J. (2006), «The effects of coal tar based pavement sealer on amphibian development and metamorphosis», Ecotoxicology, 15, 241 – 247.

Burkhart J. G., Helgen J. C., Fort D. J., Gallagher K., Bowers D., Propst T. L., Gernes M., Magner J., Shelby M. D., and Lucier G. (1998), «Induction of mortality and malformation in Xenopus laevis embryos by water sources associated with field frog deformities», Environ. Health Persp., 106, 841 – 848.

Burlibasa L. and Gavrila L. (2010), «Amphibians as model organisms for study environmental genotoxicity», Appl. Ecol. Environ. Res., 9, 1 – 15.

Calevro F., Campani S., Ragghianti M., Bucci S., and Mancino G. (1998), «Tests of toxicity and teratogenicity in biphasic vertebrates treated with heavy metals (Cr3+, Al3+, Cd2+)», Chemosphere, 37, 3011 – 3017.

Carr J. A., Gentles A., Smith E. E., Goleman W. L., Urguidi L. J., Thuett K., Kendall R. J., Giesy J. P., Gross T. S., Solomon K. R., and Van Der Kraak G. (2003), «Response of larval Xenopus laevis to atrazine: assessment of growth, metamorphosis and gonadal and laryngeal morphology», Environ. Toxicol. Chem., 22, 396 – 405.

Conrad J. (2010), «Amphibians», Retrieved from The Backyard Nature Website at amphibs.htm.

Cooke A. S. (1972), «Indications of recent changes in the status in the British Isles of the frog (Rana temporaria) and the toad (Bufo bufo)», J. Zool. Lond., 167, 161 – 178.

Corn P. S. (1994), «What we know and don’t know about amphibian declines in the west», in: W. W. Covington and L. F. DeBano (technical coordinators), Sustainable Ecological Systems: Implementing as Ecological Approach to Land Management. General Technical Reports. RM-247, Ft. Collins, CO, USDA Forest Service Rocky Mountain Forest Range Experimental Station.

Dabagyan N. V. and Sleptzova L. A. (1975), «Common frog (Rana temporaria L.)», in: Objects in Developmental Biology [in Russian], Nauka.

Demichelis S. O., De La Torre F. R., Ferrari L., Garcia M. E., and Salibian A. (2001), «Tadpoles assay: its application to a water toxicity assessment of polluted urban river», Environ. Monit. Assess., 68, 63 – 73.

Dunson W. A. and Connell J. (1982), «Specific inhibitor of hatching in amphibian embryos by low pH», J. Herpetol., 16, 314 – 316.

Elsdale T. and Davidson D. (1983), «Somitogenesis in amphibia IV. The dynamics of tail development», J. Embryol. Exp. Morphol., 76, 157 – 176.

Elsdale T., Pearson M., and Whitehead M. (1976), «Abnormalities in somite segmentation following heat shock to Xenopus embryos», J. Embryol. Exp. Morphol., 35, 625 – 635.

Fort D. J., Propst T. L., Stover E. L, Helgen J. C., Levey R. B., Gallagher K., and Burkhart J. B. (1999a), «Effects of pond water, sediment, and sediment extracts from Minnesota and Vermont, USA, on early development and metamorphosis of Xenopus», Environ. Toxicol. Chem., 18, 2305 – 2315.

Fort D. J., Rogers R. L., Copley H. F., Bruning L. A., Stover E. L., Helgen J. C., and Burkhart J. B. (1999b), «Progress toward identifying causes of maldevelopment induced in Xenopus by pond water and sediment extracts from Minnesota, USA», Environ. Toxicol. Chem., 18, 2316 – 2324.

Fort D. J., Rogers R. L., Paul R. P., Miller M. F., Clark P., Stover E. L., Yoshioko J., Quimby F., Sower S. A., Reed K. L., Babbitt K. J., and Rolland R. (2001), «Effects of pond water, sediment and sediment extract samples from New Hampshire USA on early Xenopus development and metamorphosis: comparison to native species», J. Appl. Toxicol., 21, 199 – 209.

Fort D. J., Rogers R. L., Thomas J. H., Buzzard B. O., Noll A. M., and Spaulding C. D. (2004), «Comparative sensitivity of Xenopus tropicalis and Xenopus laevis as test species for the FETAX model», J. Appl. Toxicol., 24, 443 – 457.

Fort D. J., Rogers R. L., Thomas J. H., Hopkins W. A., and Schlekat C. (2006), «Comparative developmental toxicity of nickel to Gastrophryne carolinensis, Bufo terrestris, and Xenopus laevis», Arch. Environ. Contam. Toxicol., 51, 703 – 710.

Freda J. and Dunson W. A. (1984), «Sodium balance of amphibian larvae exposed to low environmental pH», Physiol. Zool., 57, 435 – 443.

Freda J. and Dunson W. A. (1985), «Field and laboratory studies of ion balance and growth rates of ranid tadpoles chronically exposed to low pH», Copeia, 1985(2), 415 – 425.

Fulton M. H. and Chambers J. E. (1985), «The toxic and teratogenic effects of selected organophosphorus compounds on the embryos of three species of amphibians», Toxicol. Lett., 26, 175 – 180.

Gale N. L., Wixton B. G., Hardie M. G., and Jennett J. C. (1973), «Aquatic organisms and heavy metals in Missouri’s new lead belt», Water Resources Bull., 9, 673 – 688.

Greulich K. and Pflugmacher S. (2003), «Differences in susceptibility of various life stages of amphibians to pesticide exposure», Aquat. Toxicol., 65, 329 – 336.

Griffis-Kyle K. L. (2007), «Sublethal effects of nitrite on eastern tiger salamander (Ambystoma tigrinum tigrinum) and wood frog (Rana sylvatica) embryos and larvae: implications for field populations», Aquat. Ecol., 41, 119 – 127.

Hayes T. B., Falso P., Gallipeau S., and Stice M. (2010), «The cause of global amphibian declines: a developmental endocrinologist’s perspective», J. Exp. Biol., 213, 921 – 933.

Heyer W. R., Rand A. S., da Cruz C. A. G., and Peixoto O. L. (1988), «Decimations, extinctions, and colonization of frog populations in southeast Brazil and their evolutionary implications», Biotropica, 20, 230 – 235.

Johnson P. T. J., Lunde K. B., Ritchie E. G., and Launer A. E. (1999), «The effect of trematode infection on amphibian limb development and survivorship», Science, 284, 802 – 804.

Johnson P. T. J., Lunde K. B., Ritchie E. G., Reaser J. K., and Launer A. E. (2001) «Morphological abnormality patterns in a California amphibian community», Herpetologica, 57, 336 – 352.

Johnson P. T. J., Lunde K. B., Zelmer D. A., and Werner J. K. (2003), «Limb deformities as an emerging parasitic disease in amphibians: evidence from museum specimens and resurvey data», Conserv. Biol., 17, 1724 – 1737.

Katagiri C. (1976), «Properties of the hatching enzyme from frog embryos», J. Exp. Zool., 193, 109 – 118.

Kloas W., Lutz I., and Einspanier R. (1999), «Amphibians as a model to study endocrine disruptors. II. Estrogenic activity of environmental chemicals in vitro and in vivo», Sci. Total Environ., 225, 59 – 68.

Kovalenko E. E. (2011), «Properties of norm and variability», Russ. J. Dev. Biol., 42, 317 – 330.

Lannoo M. J. (1998), «Amphibian conservation and wetland management in the upper Midwest: a catch-22 for the cricket frog?», in: Status and Conservation of Midwestern Amphibians, pp. 330 – 339.

Laposata M. M. and Dunson W. A. (1998), «Effects of boron and nitrate on amphibian egg hatching success», Arch. Environ. Contam. Toxicol., 35, 615 – 619..

Levengood W. C. (1969), «A new teratogenic agent applied to amphibian embryos», J. Embryol. Exp. Morphol., 21, 23 – 31.

Loewengart G. (2001), «Toxicity of boron to rainbow trout: a weight-of-the-evidence assessment», Environ. Toxicol. Chem., 20, 796 – 803.

Mann R. M. and Bidwell J. R. (2000), «Application of the FETAX protocol to assess the developmental toxicity of nonylphenol ethoxylate to Xenopus laevis and two Australian frogs», Aquat. Toxicol., 51, 19 – 29.

Marques S. M., Antunes S. C., Pissarra H., Pereira M. L., Goncalves F., and Pereira R. (2009), «Hiatopathological changes and erythrocytic nuclear abnormalities in Iberian green frogs (Rana perezi Seoane) from a uranium mine pond», Aquat. Toxicol., 91, 187 – 195.

Marques S. M., Goncalves F., and Pereira R. (2008), «Effects of a uranium mine effluent in the early-life stages of Rana perezi Seoane», Sci. Total Environ., 402, 29 – 35.

McDonald D. G., Ozog J. L., and Simons B. P. (1984), «The influence of low pH environments on ion regulation in the larval stages of the anuran amphibian, Rana clamitans», Can. J. Zool., 62, 2171 – 2177.

Menon J., Gardner E. E., and Vali S. (2000), «Developmental implication of differential effects of calcium in tail and body skin of anuran tadpoles», J. Morphol., 244, 31 – 43.

Mitchell S. E., Caldwell C. A., Gonzales G., Gould W. R., and Arimoto R. (2005), «Effects of depleted uranium on survival, growth, and metamorphosis in the African clawed frog (Xenopus laevis)», J. Toxicol. Environ. Health. A, 68, 951 – 965.

Morgan M. K. (1996), «Teratogenic potential of Atrazine and 2,4-D using FETAX», J. Toxicol. Environ. Health, 28, 151 – 168.

Nixdorf W. L., Taylor D. H., and Isaacson L. G. (1997), «Use of bullfrog tadpoles (Rana catesbeiana) to examine the mechanisms of lead neurotoxicity», Am. Zool., 37, 363 – 368.

Opitz R., Braunbeck T., Bogi C., Pickford D. B., Nentwig G., Oehlmann J., Tooi O., Lutz I., and Kloas W. (2005), «Description and initial evaluation of a Xenopus metamorphosis assay for detection of thyroid system-disrupting activities of environmental compounds», Environ. Toxicol. Chem., 24, 653 – 664.

Ortiz-Santaliestra M. E., Marco A., and Lizana M. (2005), «Sensitivity and behavior of the iberian newt, Triturus boscai, under terrestrial exposure to ammonium nitrate», Bull. Environ. Contam. Toxicol., 75, 662 – 669.

Ouellet M. (2000), «Amphibian deformities: current state of Knowledge», in: D. W. Sparling, C. Bishop, and G. Linder (eds.), Ecotoxicology of Amphibians and Reptiles, pp. 617 – 661.

Owens K. D. and Baer K. N. (2000), «Modifications of the topical japanese medaka (Oryzias latipes) embryo larval assay for assessing developmental toxicity of pentachlorophenol and p,p’-dichlorodiphenyltrichloroethane», Ecotoxicol. Environ. Safety, 47, 87 – 95.

Pina H., Pekkonen M., and Merila J. (2006), «Morphological abnormalities in amphibians habitats: case study of the common frog Rana temporaria», Copeia, 2006(4), 810 – 817.

Plowman M. C., Grbac-Ivankovic S., Martin J., Hopfer S. M., and Sunderman F. W. (1994), «Malformations persist after metamorphosis of Xenopus laevis tadpoles exposed to Ni2+, Co2+, or Cd2+ in FETAX assays», Teratogen. Carcinogen. Mutagen., 14, 135 – 144.

Pough F. H. and Wilson R. E. (1977), «Acid precipitation and reproductive success of Ambystoma salamanders», Water Air Soil Pollution, 7, 531 – 544.

Read J. L. and Tyler M. J. (1990), «The nature and incidence of post-axial, skeletal abnormalities in the frog Neobatrachus centralis Parker at Olympic dam, South Australia», Transact. Roy. Soc. S. Australia, 114, 213 – 217.

Reeves M. K., Jensen P., Dolph C. L., Holyoak M., and Trust K. A. (2010), «Multiple stressors and the cause of amphibian abnormalities», Ecol. Monogr., 80, 423 – 440.

Rohr J. R. and Crumrine P. W. (2005), «Effects of an herbicide and an insecticide on pond community structure and processes», Ecol. Appl., 15, 1135 – 1147.

Romansic J. M., Waggener A. M., Bancroft B. A., and Blaustein A. R. (2009), «Influence of ultraviolet-B radiation on growth, prevalence of deformities, and susceptibility to predation in cascades frog (Rana cascadae) larvae», Hydrobiologia, 624, 219 – 233.

Ruiz A. M., Maerz J. C., Davis A. K., Keel M. K., Ferreira A. R., Conroy M. J., Morris L. A., and Fisk A. T. (2010), «Patterns of development and abnormalities among tadpoles in a constructed wetland receiving treated wastewater», Environ. Sci. Technol., 44, 4862 – 4868.

Rzehak K., Maryańska-Nadachowska A., and Jordan M. (1977), «The effect of Karbatox 75 a Carbaryl insecticide upon the development of tadpoles of Rana temporaria L. and Xenopus laevis D»., Folia Biol., 25, 391 – 399.

Schuytema G. S. and Nebeker A. V. (1998), «Comparative toxicity of diuron on survival and growth of pacific treefrog, bullfrog, red-legged frog, and African clawed frog embryos and tadpoles», Arch. Environ. Contam. Toxicol., 34, 370 – 376.

Selvi M., Gul A., and Yilmaz M. (2003), «Investigation of acute toxicity of cadmium chloride (CdCl2 • H2O) metal salt and behavioural changes it causes on water frog (Rana ridibunda Pallas, 1771)», Chemosphere, 52, 259 – 263.

Severtsova E. A. (2005), «Adaptation of Amphibian gastrulating embryos to an influence of anthropogenic pollution», in: 10th Congr. of the Eur. Soc. for Evol. Biol., Krakow, Poland.

Severtsova E. A. and Severtsov A. S. (2007), «Mechanisms of adaptive regulation of early development of Anuran amphibians dwelling the anthropogenically polluted water bodies», Zh. Obshch. Biol., 68, 323 – 31.

Sharma B. and Patiño R. (2008), «Exposure of Xenopus laevis tadpoles to cadmium reveals concentration-dependent bimodal effects on growth and monotonic effects on development and thyroid gland activity», Toxicol. Sci., 105, 51 – 58.

Snawder J. E. and Chambers J. E. (1989), «Toxic and developmental effects of organophosphorus insecticides in embryos of the South African clawed frog», J. Environ. Sci. Health B, 24, 205 – 218.

Snawder J. E. and Chambers J. E. (1990), «Critical time periods and the effect of tryptophan in malathion induced developmental defects in Xenopus embryos», Life Sci., 46, 1635 – 1642.

Sparling D. W., Krest S., and Ortiz-Santaliestra M. (2006), «Effects of lead-contaminated sediment on Rana sphenocephala tadpoles», Arch. Environ. Contam. Toxicol., 51, 458 – 466.

Sparling D. W., Linder G., and Bishop C. A. (2006), Eds. Pensacola, Society of Environmental Toxicology and Chemistry (SETAC).

Surova G. S. (2002), «Effect of acid environmental condition on viability of the common frog (Rana temporaria) eggs», Zool. Zh., 81, 608 – 616.

Surova G. S. and Severtsov A. S. (1985), «The death of the grass frog (Rana temporaria L.) in early ontogenesis and factors causing it», Russ. J. Zool., 44, 61 – 71.

Taylor B., Skelly D., Demarchis L. K., Slade M. D., Galusha D., and Rabinowitz P. M. (2005), «Proximity to pollution sources and risk of amphibian limb malformation», Environ. Health Persp., 113, 1497 – 1501.

Tyler M. J. (1989), Australian Frogs, Viking O’Neill, Melbourne.

Ulger M. (2003), Fundaments in Ecotoxicology. Second Edition, Lewis Publisher.

Vershinin V. L. (1989), «Morphological abnormalities of amphibians of the urban environment», Ross. Zh. Ékol., 3, 58 – 65.

Vershinin V. L. (1997), Ecological Features of Amphibian Populations in Urban Areas. Author’s Abstract of Doctoral Thesis, Yekaterinburg: Institute of Plant and Animal Ecology [in Russian], Ural Branch, Russian Academy of Sciences.

Vismara C., Battista V., Vailati G., and Bacchetta R. (2000), «Paraquat induced embryotoxicity on Xenopus laevis development», Aquat. Toxicol., 49, 171 – 179.

Wake D. B and Vredenburg V. T. (2008), «Are we in the midst of the sixth mass extinction? A view from the world of amphibians», PNAS, 105, 11466 – 11473.

Weinstein D. C. and Hemmati-Brivanlou A. (1999), «Neural Induction», Annu. Rev. Cell Dev. Biol., 15, 411 – 433.

Werner E. E. (1991), «Nonlethal effects of a predator on competitive interaction between two anuran larvae», Ecology, 72, 1790 – 1720.

Werner E. E. and Bradley R. A. (1996), «Predator-induced behavioral indirect effects: consequences to competitive interactions in anuran larvae», Ecology, 77, 157 – 169.

Yong-Uk P., Chun-Sik Y., Jong-Hyang K., Joo-Hung P., and Seon-Woo C. (2010), «Numerical variations and spontaneous malformations in the early embryos of the Korean salamander, Hynobius leechii, in the farmlands of Korea», Environ. Toxicol., 25, 533 – 544.

Zamaletdinov R. I. (2003), «The morphological anomalies in urban populations of anurans (in the city of Kazan’)», Curr. Studies Herpetol., 2, 148 – 153.



  • There are currently no refbacks.