, Volume 22, Issue 1, pp 125–138 | Cite as

Experimental exposure of red-legged partridges (Alectoris rufa) to seeds coated with imidacloprid, thiram and difenoconazole

  • Ana Lopez-Antia
  • Manuel E. Ortiz-SantaliestraEmail author
  • François Mougeot
  • Rafael Mateo


Pesticide coated seeds are commonly used in agriculture, and may be an important source of food for some birds in times of scarcity, as well as a route of pesticide ingestion. We tested the lethal and sub-lethal effects of treated seed ingestion by the red-legged partridge (Alectoris rufa), a game bird of high socio-economic value in Spain. One year-old partridges (n = 42 pairs) were fed for 10 days in spring (prior to breeding) with wheat treated with difenoconazole (fungicide), thiram (fungicide) or imidacloprid (insecticide), using two doses for each pesticide (the one recommended, and its double to represent potential cases of abuse of pesticides). We investigated the direct and indirect effects on the body condition, physiology, immunology, coloration and subsequent reproduction of exposed partridges. For the latter, eggs were collected, measured and incubated and the growth and survival of chicks were monitored. Thiram and imidacloprid at high exposure doses produced mortalities of 41.6 and 58.3 %, respectively. The first death was observed at day 3 for imidacloprid and at day 7 for thiram. Both doses of the three pesticides caused sublethal effects, such as altered biochemical parameters, oxidative stress and reduced carotenoid-based coloration. The high exposure doses of imidacloprid and thiram also produced a decrease in cellular immune response measured by the phytohemagglutinin test in males. Bearing in mind the limitation of the small number of surviving pairs in some treatments, we found that the three pesticides reduced the size of eggs and imidacloprid and difenoconazole also reduced the fertilization rate. In addition, both thiram and imidacloprid reduced chick survival. These experiments highlight that the toxicity of pesticide-treated seeds is a factor to consider in the decline of birds in agricultural environments.


Breeding success Carotenoid-dependent coloration Fungicide Immune response Insecticide Oxidative stress 



Study financed by FEDENCA (Real Federación Española de Caza) and Oficina Nacional de la Caza with the partnership of Fundación Biodiversidad. We thank Pablo Camarero, Jaime Rodríguez-Estival and Xurxo Piñeiro for their help.


  1. Alonso-Alvarez C, Ferrer M, Velando A (2002) The plasmatic index of body condition in yellow-legged Gulls Larus cachinnans: a food-controlled experiment. Ibis 144:147–149CrossRefGoogle Scholar
  2. Alonso-Alvarez C, Pérez-Rodríguez L, Mateo R, Chastel O, Viñuela J (2008) The oxidation handicap hypothesis and the carotenoid allocation trade-off. J Evol Biol 21:1789–1797CrossRefGoogle Scholar
  3. Alonso-Alvarez C, Pérez-Rodríguez L, Garcia JT, Viñuela J (2009) Testosterone-mediated trade-offs in the old age: a new approach to the immunocompetence handicap and carotenoid-based sexual signalling. Proc R Soc B Biol Sci 276:2093–2101CrossRefGoogle Scholar
  4. Avery ML, Decker DG, Fischer DL, Stafford TR (1993) Responses of captive blackbirds to a new insecticidal seed treatment. J Wildl Manag 57:652–656CrossRefGoogle Scholar
  5. Aydin B (2011) Effects of thiacloprid, deltamethrin and their combination on oxidative stress in lymphoid organs, polymorphonuclear leukocytes and plasma of rats. Pestic Biochem Physiol 100:165–171CrossRefGoogle Scholar
  6. Balani T, Agrawal S, Thaker AM (2011) Hematological and biochemical changes due to short-term oral administration of imidacloprid. Toxicol Int 18:2–4CrossRefGoogle Scholar
  7. Banerjee BD, Seth V, Ahmed RS (2001) Pesticide-induced oxidative stress: perspectives and trends. Rev Environ Health 16:1–40CrossRefGoogle Scholar
  8. Berthouly A, Helfenstein F, Richner H (2007) Cellular immune response, stress resistance and competitiveness in nestling great tits in relation to maternally transmitted carotenoids. Funct Ecol 21:335–343CrossRefGoogle Scholar
  9. Bhardwaj S, Srivastava MK, Kapoor U, Srivastava LP (2010) A 90 days oral toxicity of imidacloprid in female rats: morphological, biochemical and histopathological evaluations. Food Chem Toxicol 48:1185–1190CrossRefGoogle Scholar
  10. Blomqvist D, Johansson OC, Götmark F (1997) Parental quality and egg size affect chick survival in a precocial bird, the lapwing Vanellus vanellus. Oecologia 110:18–24CrossRefGoogle Scholar
  11. Bortolotti GR, Negro JJ, Surai PF, Prieto P (2003) Carotenoids in eggs and plasma of red-legged partridges: effects of diet and reproductive output. Physiol Biochem Zool 76:367–374CrossRefGoogle Scholar
  12. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254CrossRefGoogle Scholar
  13. Bro E, Decors A, Millot F, Soyez D, Moinet M, Berny P, Mastain O (2010) Intoxications des perdrix grises en nature. Nouveau bilan de la surveillance «SAGIR». Faune Sauvag 289:26–32Google Scholar
  14. Cabezas-Díaz S, Virgós E (2007) Adaptive and non-adaptive explanations for hatching failure in eggs of the red-legged partridge Alectoris rufa. Ardea 95:55–63CrossRefGoogle Scholar
  15. Costantini D (2008) Oxidative stress in ecology and evolution: lessons from avian studies. Ecol Lett 11:1238–1251Google Scholar
  16. Cucco M, Guasco B, Malacarne G, Ottonelli R, Tanvez A (2008) Yolk testosterone levels and dietary carotenoids influence growth and immunity of grey partridge chicks. Gen Comp Endocrinol 156:418–425CrossRefGoogle Scholar
  17. Dacke CG (2000) The parathyroids, calcitonin and vitamin D. In: Whittow GC (ed) Sturkie’s avian physiology. Academic Press, San Diego, pp 473–488CrossRefGoogle Scholar
  18. de Snoo GR, Scheidegger NMI, de Jong FMW (1999) Vertebrate wildlife incidents with pesticides: a European survey. Pest Manag Sci 55:47–54Google Scholar
  19. Edwards HM Jr (1987) Effects of thiuram, disulfiram and a trace element mixture on the incidence of tibial dyschondroplasia in chickens. J Nutr 117:964–969Google Scholar
  20. EL-Gendy KS, Aly NM, Mahmoud FH, Kenawy A, El-Sebae AKH (2010) The role of vitamin C as antioxidant in protection of oxidative stress induced by imidacloprid. Food Chem Toxicol 48:215–221CrossRefGoogle Scholar
  21. Galloway T, Handy R (2003) Immunotoxicity of organophosphorous pesticides. Ecotoxicology 12:345–363CrossRefGoogle Scholar
  22. Gatne MM, Ramesh Bhoir PS, Deore MD (2006) Immunotoxicity studies of imidacloprid in rats. Toxicol Int 13:89–92Google Scholar
  23. Geiger F, Bengtsson J, Berendse F, Weisser WW, Emmerson M, Morales MB, Ceryngier P, Liira J, Tscharntke T, Winqvist C, Eggers S, Bommarco R, Pärt T, Bretagnolle V, Plantegenest M, Clement LW, Dennis C, Palmer C, Oñate JJ, Guerrero I, Hawro V, Aavik T, Thies C, Flohre A, Hänke S, Fischer C, Goedhart PW, Inchausti P (2010) Persistent negative effects of pesticides on biodiversity and biological control potential on European farmland. Basic Appl Ecol 11:97–105CrossRefGoogle Scholar
  24. Giraudeau M, Duval C, Czirják GÁ, Bretagnolle V, Eraud C, McGraw KJ, Heeb P (2011) Maternal investment of female mallards is influenced by male carotenoid-based coloration. Proc R Soc B Biol Sci 278:781–788CrossRefGoogle Scholar
  25. Goto N, Kodama H, Okada K, Fujimoto Y (1978) Suppression of phytohemagglutinin skin response in thymectomized chickens. Poult Sci 57:246–250CrossRefGoogle Scholar
  26. Griffiths R, Double MC, Orr K, Dawson RJ (1998) A DNA test to sex most birds. Mol Ecol 7:1071–1075CrossRefGoogle Scholar
  27. Grosicka-Maciąg E, Kurpios D, Czeczot H, Szumiło M, Skrzycki M, Suchocki P, Rahden-Staroń I (2008) Changes in antioxidant defense systems induced by thiram in V79 Chinese hamster fibroblasts. Toxicol In Vitro 22:28–35CrossRefGoogle Scholar
  28. Guitart R, Mateo R, Gutierrez JM, To-Figueras J (1996) An outbreak of thiram poisoning on Spanish poultry farms. Vet Hum Toxicol 38:287–288Google Scholar
  29. Hart ADM (1990) The assessment of pesticide hazards to birds: the problem of variable effects. Ibis 132:192–204CrossRefGoogle Scholar
  30. Hudson RH, Tucker RK, Haegele MA (1984) Handbook of toxicity of pesticides to wildlife. Resource publication 153. United States Department of the Interior, Fish and Wildlife Service, Washington, DCGoogle Scholar
  31. Kapoor U, Srivastava MK, Bhardwaj S, Srivastava LP (2010) Effect of imidacloprid on antioxidant enzymes and lipid peroxidation in female rats to derive its No Observed Effect Level (NOEL). J Toxicol Sci 35:577–581CrossRefGoogle Scholar
  32. KEM (2006) Draft assessment report. Difenoconazole, vol 3. Swedish Chemicals Inspectorate, SundbybergGoogle Scholar
  33. Knight JA (2000) Review: free radicals, antioxidants, and the immune system. Ann Clin Lab Sci 30:145–158Google Scholar
  34. Korhonen A, Hemminki K, Vainio H (1982) Application of the chicken embryo in testing for embryotoxicity. Thiurams. Scand J Work Environ Health 8:63–69CrossRefGoogle Scholar
  35. Lewandowski AH, Campbell TW, Harrison GJ (1986) Clinical chemistries. In: Harrison GJ, Harrison LR (eds) Clinical avian medicine and surgery. W.B. Saunders Company, Philadelphia, pp 192–200Google Scholar
  36. Li J, Bi D, Pan S, Zhang Y (2007) Effect of diet with thiram on liver antioxidant capacity and tibial dyschondroplasia in broilers. Br Poult Sci 48:724–728CrossRefGoogle Scholar
  37. Marikovsky M (2002) Thiram inhibits angiogenesis and slows the development of experimental tumours in mice. Br J Cancer 86:779–787CrossRefGoogle Scholar
  38. Mineau P (2005) A review and analysis of study endpoints relevant to the assessment of “long term” pesticide toxicity in avian and mammalian wildlife. Ecotoxicology 14:775–799CrossRefGoogle Scholar
  39. MMAMRM (2010) Registro de productos fitosanitarios. Ministerio de Medio Ambiente y Medio Rural y Marino. Accessed 06 Mar 2010
  40. Mougeot F, Pérez-Rodríguez L, Sumozas N, Terraube J (2009) Parasites, condition, immune responsiveness and carotenoid-based ornamentation in male red-legged partridge Alectoris rufa. J Avian Biol 40:67–74CrossRefGoogle Scholar
  41. Muthukumaran S, Sudheer AR, Menon VP, Nalini N (2008) Protective effect of quercetin on nicotine-induced prooxidant and antioxidant imbalance and DNA damage in Wistar rats. Toxicology 243:207–215CrossRefGoogle Scholar
  42. Peig J, Green AJ (2009) New perspectives for estimating body condition from mass/length data: the scaled mass index as an alternative method. Oikos 118:1883–1891CrossRefGoogle Scholar
  43. Prosser PJ, Hart ADM, Langton SD, McKay HV, Cooke AS (2006) Estimating the rate of poisoning by insecticide-treated seeds in a bird population. Ecotoxicology 15:657–664CrossRefGoogle Scholar
  44. Pruett SB, Barnes DB, Han YC, Munson AE (1992) Immunotoxicological characteristics of sodium methyldithiocarbamate. Fundam Appl Toxicol 18:40–47CrossRefGoogle Scholar
  45. Reglero MM, Taggart MA, Monsalve-González L, Mateo R (2009) Heavy metal exposure in large game from a lead mining area: effects on oxidative stress and fatty acid composition in liver. Environ Pollut 157:1388–1395CrossRefGoogle Scholar
  46. Rodríguez-Estival J, Martínez-Haro M, Martín-Hernando MP, Mateo R (2010) Sub-chronic effects of nitrate in drinking water on red-legged partridge (Alectoris rufa): oxidative stress and T-cell mediated immune function. Environ Res 110:469–475CrossRefGoogle Scholar
  47. Siddiqui A, Choudhary M, Goriya HV, Bhavsar SK, Thaker AM (2007) Evaluation of immunotoxic effect of short-term administration of quinalphos and imidacloprid in white leghorn cockerels. Toxicol Int 14:15–19Google Scholar
  48. Stanley PI, Bunyan PJ (1979) Hazards to wintering geese and other wildlife from the use of dieldrin, chlorfenvinphos and carbophenothion as wheat seed treatments. Proc R Soc Lond B Biol Sci 205:31–45CrossRefGoogle Scholar
  49. Subapriya S, Vairamuthu S, Manohar BM, Balachandran C (2007) Growth performance studies in thiram toxicosis in broiler chicken. Int J Poult Sci 6:248–250CrossRefGoogle Scholar
  50. Tomlin CDS (2004/05) Imidacloprid (138261-41-3). In: The e-pesticide manual, version 3.1, 13th edn. British Crop Protection Council, SurreyGoogle Scholar
  51. Villafuerte R, Negro JJ (1998) Digital imaging for colour measurement in ecological research. Ecol Lett 1:151–154CrossRefGoogle Scholar
  52. Voccia I, Blakley B, Brousseau P, Fournier M (1999) Immunotoxicity of pesticides: a review. Toxicol Ind Health 15:119–132CrossRefGoogle Scholar
  53. Werner SJ, Linz GM, Tupper SK, Carlson JC (2010) Laboratory efficacy of chemical repellents for reducing blackbird damage in rice and sunflower crops. J Wildl Manag 74:1400–1404Google Scholar
  54. Wilson JD, Morris AJ, Arroyo BE, Clark SC, Bradbury RB (1999) A review of the abundance and diversity of invertebrate and plant foods of granivorous birds in northern Europe in relation to agricultural change. Agric Ecosyst Environ 75:13–30CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2012

Authors and Affiliations

  • Ana Lopez-Antia
    • 1
  • Manuel E. Ortiz-Santaliestra
    • 1
    Email author
  • François Mougeot
    • 2
  • Rafael Mateo
    • 1
  1. 1.Instituto de Investigación en Recursos Cinegéticos (IREC), CSIC-UCLM-JCCMCiudad RealSpain
  2. 2.Estación Experimental de Zonas Áridas (EEZA), CSICAlmeríaSpain

Personalised recommendations