Abstract
Pollution was cited by the Global Amphibian Assessment to be the second most important cause of amphibian decline worldwide, however, the effects of the agricultural environment on amphibians are not well understood. In this study, spawn from Bufo bufo was taken from four sites in England and Wales with varying intensities of arable agriculture. Spawn was either placed in tanks containing aged tap water (ex-situ, five replicates) or in cages at the native site (caged, five replicates). Hatching success, abnormal tadpoles, and forelimb emergence were recorded during the larval stage. Individuals were also sampled at five time points (TP) during development (5-, 7-, 9-, 12-, 15-weeks post-hatch) and analysed for morphological parameters. The thyroids (TP2) and the gonads (TP3,4,5) were also analysed histologically. With the exception of the thyroid histopathology, all analysed endpoints were significantly different between ex-situ individuals reared under identical conditions from the different sites. In addition, intensity of arable agriculture had a negative effect on growth and development. At one site, despite distinct rearing conditions, a high level of intersex (up to 42%) and similar sex ratios were observed in both ex-situ and caged individuals. Taken together, these data suggest that maternal exposure and/or events in ovo had a much larger effect on growth, metamorphic development, and sexual differentiation in B. bufo than the ambient environment. This could have important implications for traditional exposure scenarios that typically begin at the larval stage. Intersex is reported for the first time in European amphibians in situ, highlighting the potential use of distinct populations of amphibians in fundamental research into the aetiology of specific developmental effects in wild amphibians.
Similar content being viewed by others
Abbreviations
- TP:
-
Time point
- SVL:
-
Snout-vent length
- HLL:
-
Hindlimb length
- FLE:
-
Forelimb emergence
- DO:
-
Dissolved oxygen
- TO:
-
Testicular oocyte
References
Beldomenico PM, Rey F, Prado WS, Villarreal JC, Munoz-de-Toro M, Luque EH (2007) In ovum exposure to pesticides increases the egg weight loss and decreases hatchlings weight of Caiman latirostris (Crocodylia: Alligatoridae). Ecotoxicol Environ Saf 68(2):246–251
Bishop CA, Mahony NA, Struger J, Ng P, Pettit KE (1999) Anuran development, density and diversity in relation to agricultural activity in the Holland River watershed, Ontario, Canada (1990–1992). Environ Monit Assess 57(1):21–43
Carr JA, Gentles A, Smith EE, Goleman WL, Urquidi LJ, Thuett K, Kendall RJ, Giesy JP, Gross TS, Solomon KR, Van der Kraak G (2003) Response of larval Xenopus laevis to atrazine: assessment of growth, metamorphosis, and gonadal and laryngeal morphology. Env Tox Chem 22(2):396–405
Calisi RM (2005) Variation in Bidder’s organ volume is attributable to reproductive status in Bufo woodhousii. J Herpetol 39(4):656–659
Cooke AS (1977) Effects of field applications of herbicides diquat and dichlobenil on amphibians. Environ Pollut 12(1):43–50
Davidson C, Knapp RA (2007) Multiple stressors and amphibian declines: dual impacts of pesticides and fish on yellow-legged frogs. Ecol Appl 17(2):587–597
DEFRA (2005) e-Digest statistics about: Inland water quality and use. http://www.defra.gov.uk/environment/statistics/inlwater/iwgroundwater.htm. Retrieved January 2007
Falconi R, Dalpiaz D, Zaccanti F (2004) Ultrastructural aspects of gonadal morphogenesis in Bufo bufo (Amphibia anura) 1. Sex differentiation. J Exp Zool A Comp Exp Biol 301A(5):378–388
Falconi R, Dalpiaz D, Zaccanti F (2007) Morphological aspects of gonadal morphogenesis in Bufo bufo (Amphibia anura): Bidder’s organ differentiation. Anat Rec Adv Integr Anat Evol Biol 290(7):801–813
Fellers GM, McConnell LL, Pratt D, Datta S (2004) Pesticides in mountain yellow-legged frogs (Rana muscosa) from the Sierra Nevada Mountains of California, USA. Environ Toxicol Chem 23(9):2170–2177
Fort DJ, Paul RR (2002) Enhancing the predictive validity of frog embryo teratogenesis assay—Xenopus (FETAX). J Appl Toxicol 22(3):185–191
GAA (2004) http://www.iucnredlist.org/initiatives/amphibians/analysis/major-threats. Retrieved Nov 2009
Greulich K, Pflugmacher S (2004) Uptake and effects on detoxification enzymes of cypermethrin and tadpoles of embryos. Arch Env Cont Tox 47(4):489–495
Gruca S, Michalowski J (1961) The rate of development of gonads against the backdrop of the development of the whole organism in the tadpole Xenopus laevis Daud. Acta Biol Cracoviensia Ser Zool 4:191–197
Gyllenhammar I, Holm L, Eklund R, Berg C (2009) Reproductive toxicity in Xenopus tropicalis after developmental exposure to environmental concentrations of ethynylestradiol. Aquat Toxicol 91(2):171–178
Harris RN (1999) The anuran tadpole: evolution and maintenance. In: McDiarmid RW, Altig R (eds) Tadpoles: the biology of anuran larvae. University of Chicago Press, Chicago, pp 279–294
Hayes TB (1995) Interdependance of corticosterone-hormones and thyroid-hormones in larval toads (Bufo boreas). 1. Thyroid hormone-dependent and hormone-independent effects of corticosterone on growth and development. J Exp Zool 271(2):95–102
Hayes TB, Collins A, Lee M, Mendoza M, Noriega N, Stuart AA, Vonk A (2002) Hermaphroditic, demasculinized frogs after exposure to the herbicide atrazine at low ecologically relevant doses. Proc Natl Acad Sci USA 99(8):5476–5480
Hayes T, Haston K, Tsui M, Hoang A, Haeffele C, Vonk A (2003) Atrazine-induced hermaphroditism at 0.1 ppb in American leopard frogs (Rana pipiens): laboratory and field evidence. Environ Health Perspect 111(4):568–575
Hayes TB, Case P, Chui S, Chung D, Haeffele C, Haston K, Lee M, Mai VP, Marjuoa Y, Parker J, Tsui M (2006) Pesticide mixtures, endocrine disruption, and amphibian declines: are we underestimating the impact? Environ Health Perspect 114:40–50
Hopkins WA, DuRant SE, Staub BP, Rowe CL, Jackson BP (2006) Reproduction, embryonic development, and maternal transfer of contaminants in the amphibian Gastrophryne carolinensis. Environ Health Perspect 114(5):661–666
Hsu CY, Hsu LH, Liang HM (1979) The effect of cyproterone acetate on the activity of delta5-3beta-hydroxysteroid dehydrogenase in tadpole sex transformation. Gen Comp Endocrinol 39:404–410
Jobling S, Coey S, Whitmore JG, Kime DE, Van Look KJW, McAllister BG, Beresford N, Henshaw AC, Brighty G, Tyler CR, Sumpter JP (2002) Wild intersex roach (Rutilus rutilus) have reduced fertility. Biol Reprod 67(2):515–524
Karasov WH, Jung RE, Vanden Langenberg S, Bergeson TLE (2005) Field exposure of frog embryos and tadpoles along a pollution gradient in the Fox River and Green Bay ecosystem in Wisconsin, USA. Environ Toxicol Chem 24(4):942–953
Kloas W, Lutz I, Urbatzka R, Springer T, Krueger H, Wolf J, Holden L, Hosmer A (2009) Does atrazine affect larval development and sexual differentiation of South African clawed frogs? Ann NY Acad Sci 1163:437–440
Knutson MG, Richardson WB, Reineke DM, Gray BR, Parmelee JR, Weick SE (2004) Agricultural ponds support amphibian populations. Ecol Appl 14(3):669–684
Larsson DGJ, Hallman H, Forlin L (2000) More male fish embryos near a pulp mill. Environ Toxicol Chem 19(12):2911–2917
Laugen AT, Laurila A, Merila J (2002) Maternal and genetic contributions to geographical variation in Rana temporaria larval life-history traits. Biol J Linn Soc 76(1):61–70
Laugen AT, Laurila A, Rasanen K, Merila J (2003) Latitudinal countergradient variation in the common frog (Rana temporaria) development rates—evidence for local adaptation. J Evol Biol 16(5):996–1005
McAllister BG, Kime DE (2004) Early life exposure to environmental levels of the aromatase inhibitor tributyltin causes masculinisation and irreversible sperm damage in zebrafish (Danio rerio) (vol 65, pg 309, 2003). Aquat Toxicol 67(3):301–302
McCoy KA, Bortnick LJ, Campbell CM, Hamlin HJ, Guillette LJ, St. Mary CM (2008) Agriculture alters gonadal form and function in the toad Bufo marinus. Environ Health Perspect 116(11):1526–1532
McDaniel TV, Martin PA, Struger J, Sherry J, Marvin CH, McMaster ME, Clarence S, Tetreault G (2008) Potential endocrine disruption of sexual development in free ranging male northern leopard frogs (Rana pipiens) and green frogs (Rana clamitans) from areas of intensive row crop agriculture. Aquat Toxicol 88(4):230–242
Nuckols JR, Ward MH, Jarup L (2004) Using geographic information systems for exposure assessment in environmental epidemiology studies. Environ Health Perspect 112(9):1007–1015
OECD (2007) Series on testing and assessment. Guidance document on amphibian thyroid histology. Environmental Health and Safety Publications, Paris, France
Ogielska M, Kotusz A (2004) Pattern and rate of ovary differentiation with reference to somatic development in anuran amphibians. J Morphol 259(1):41–54
Ohtani H, Miura I, Ichikawa Y (2000) Effects of dibutyl phthalate as an environmental endocrine disruptor on gonadal sex differentiation of genetic males of the frog Rana rugosa. Environ Health Perspect 108(12):1189–1193
Orton F, Handy RD, Carr JA (2006) Effects of nitrate and atrazine on larval development and sexual differentiation in the northern leopard frog, Rana pipiens. Environ Toxicol Chem 25(1):65–71
Orton F, Lutz I, Kloas W, Routledge EJ (2009) Endocrine disrupting effects of herbicides and pentachlorophenol: in vitro and in vivo evidence. Environ Sci Technol 43(6):2144–2150
Petrini S, Zaccanti F (1998) The effects of aromatase and 5 alpha-reductase inhibitors, antiandrogen, and sex steroids on Bidder’s organs development and gonadal differentiation in Bufo bufo tadpoles. J Exp Zool 280(3):245–259
Relyea RA, Diecks N (2008) An unforeseen chain of events: lethal effects of pesticides on frogs at sublethal concentrations. Ecol Appl 18(7):1728–1742
Rohr JR, Raffel TR, Sessions SK, Hudson PJ (2008) Understanding the net effects of pesticides on amphibian trematode infections. Ecol Appl 18(7):1743–1753
Rose CS (2005) Integrating ecology and developmental biology to explain the timing of frog metamorphosis. Trends Ecol Evol 20(3):129–135
Rugh R (1951) The frog: its reproduction and development. McGraw-Hill Book Company, Inc, New York
Smith EE, Du Preez LH, Gentles A, Solomon KR, Tandler B, Carr JA, Van der Kraak GL, Kendall RJ, Giesy JP, Gross TS (2005) Assessment of laryngeal muslce and testicular cell types in Xenopus laevis (Anura Pipidae) inhabiting maize and non-maize growing areas of South Africa. Afr J Herpet 54(1):69–76
Sparling DW, Fellers GM, McConnell LL (2001) Pesticides and amphibian population declines in California, USA. Environ Toxicol Chem 20(7):1591–1595
Witschi E (1929) Studies on sex differentiation and sex determination in amphibians. I. Development and sexual differentiation of the gonads of Rana sylvatica. J Exp Zool 52(2):235–265
Zaccanti F, di Grande F, Pasquini CP (1969) Effeti del testosterone sulle gonadi e sugli organi di Bidder in esemplari postmetamorfici di Bufo bufo. Academic Nazionale del Lincei Series 8 46:106–110
Zerulla M, Lange R, Steger-Hartmann T, Panter G, Hutchinson T, Dietrich DR (2002) Morphological sex reversal upon short-term exposure to endocrine modulators in juvenile fathead minnow (Pimephales promelas). Toxicol Lett 131(1–2):51–63
Acknowledgments
Funding from DEFRA gratefully acknowledged. Also, the generous sharing of amphibian databases from Froglife (registered charity: 1093372).
Author information
Authors and Affiliations
Corresponding author
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Orton, F., Routledge, E. Agricultural intensity in ovo affects growth, metamorphic development and sexual differentiation in the Common toad (Bufo bufo). Ecotoxicology 20, 901–911 (2011). https://doi.org/10.1007/s10646-011-0658-5
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10646-011-0658-5