Eur. J. Entomol. 110 (4): 567-576, 2013 | DOI: 10.14411/eje.2013.077

Molecular analysis of the gut contents of Harmonia axyridis (Coleoptera: Coccinellidae) as a method for detecting intra-guild predation by this species on aphidophagous predators other than coccinellids

Brecht INGELS1, Alexandre AEBI2,3, Louis HAUTIER4, Thomas VAN LEEUWEN1, Patrick DE CLERCQ1
1 Laboratory of Agrozoology, Department of Crop Protection, Faculty of Bioscience Engineering, Ghent University, Coupure links 653, 9000 Ghent, Belgium; e-mails: brecht.ingels@ugent.be; thomas.vanleeuwen@ugent.be; patrick.declercq@ugent.be
2 Agroscope Reckenholz-Tšnikon, Research Station ART, Reckenholzstrasse 191, 8046 ZŁrich, Switzerland; e-mail: alexandre.aebi@unine.ch
3 Laboratory of Soil Biology, University of Neuch‚tel, Rue Emile-Argand 11, 2000 Neuch‚tel, Switzerland
4 Biological Control and Plant Genetic Resources Department, Walloon Agricultural Research Centre, Rue de Liroux 2, 030 Gembloux, Belgium; e-mail: hautier@cra.wallonie.be

Several studies have demonstrated that the invasive ladybird Harmonia axyridis is a strong intra-guild predator of native species of ladybird. Laboratory studies have shown that H. axyridis can be an intra-guild predator of aphid predators other than coccinellids, including the hoverfly Episyrphus balteatus and lacewing Chrysoperla carnea. However, little is known about the effect of intra-guild predation (IGP) by H. axyridis on hoverfly and lacewing populations in the field. In the present study molecular analyses were used to detect the DNA of E. balteatus and C. carnea in the gut contents of H. axyridis. Primers for the syrphid and chrysopid prey were designed and feeding experiments performed to determine how long prey DNA remains detectable in the guts of this ladybird. DNA detection was influenced by the life stage of the predator and species of prey. Meal size did not affect detection time, except when fourth instar individuals of H. axyridis were fed 10 eggs or one second instar of C. carnea. Predator weight, sex and morpho-type (melanic/non-melanic) did not influence DNA detection. The half-life of the time for which the DNA of the prey remained detectable was calculated for each predator-prey combination, and ranged from 8.9 to 52.4 h. This method can be used to study the ecological importance of IGP by H. axyridis on aphidophagous predators other than coccinellids in the field.

Keywords: Coleoptera, Coccinellidae, Harmonia axyridis, Neuroptera, Chrysopidae, Chrysoperla carnea, Diptera, Syrphidae, Episyrphus balteatus, intraguild predation, gut-content analysis

Received: November 5, 2012; Accepted: April 8, 2013; Prepublished online: October 1, 2013; Published: December 1, 2013

Download citation

References

  1. AEBI A., BROWN P.M.J., DE CLERCQ P., HAUTIER L., HOWE A., INGELS B., RAVN H.P., SLOGGETT J.J., ZINDEL R. & THOMAS A. 2011: Detecting arthropod intraguild predation in the field. - BioControl 56: 429-440 Go to original source...
  2. AGUSTI N., SHAYLER S.P., HARWOOD J.D., VAUGHAN I.P., SUNDERLAND K.D. & SYMONDSON W.O.C. 2003: Collembola as alternative prey sustaining spiders in arable ecosystems: Prey detection within predators using molecular markers. - Mol. Ecol. 12: 3467-3475 Go to original source...
  3. ALHMEDI A., HAUBRUGE E. & FRANCIS F. 2009: Effect of stinging nettle habitats on aphidophagous predators and parasitoids in wheat and green pea fields with special attention to the invader Harmonia axyridis Pallas (Coleoptera: Coccinellidae). - Entomol. Sci. 12: 349-358 Go to original source...
  4. ALHMEDI A., HAUBRUGE E. & FRANCIS F. 2010: Intraguild interactions and aphid predators: Biological efficiency of Harmonia axyridis and Episyrphus balteatus. - J. Appl. Entomol. 134: 34-44 Go to original source...
  5. ARIM M. & MARQUET P.A. 2004: Intraguild predation: A widespread interaction related to species biology. - Ecol. Lett. 7: 557-564 Go to original source...
  6. BLANKENSHIP L.E. & YAYANOS A.A. 2005: Universal primers and PCR of gut contents to study marine invertebrate diets. - Mol. Ecol. 14: 891-899 Go to original source...
  7. BOWLES J. & MCMANUS D.P. 1993: Rapid discrimination of Echinococcus species and strains using a polymerase chain reaction-based rflp method. - Mol. Biochem. Parasit. 57: 231-240 Go to original source...
  8. BROWN P.M.J., FROST R., DOBERSKI J., SPARKS T., HARRINGTON R. & ROY H.E. 2011a: Decline in native ladybirds in response to the arrival of Harmonia axyridis: Early evidence from england. - Ecol. Entomol. 36: 231-240 Go to original source...
  9. BROWN P.M.J., THOMAS C.E., LOMBAERT E., JEFFRIES D.L., ESTOUP A. & HANDLEY L.J.L. 2011b: The global spread of Harmonia axyridis (Coleoptera: Coccinellidae): Distribution, dispersal and routes of invasion. - BioControl 56: 623-641 Go to original source...
  10. CHEN Y., GILES K.L., PAYTON M.E. & GREENSTONE M.H. 2000: Identifying key cereal aphid predators by molecular gut analysis. - Mol. Ecol. 9: 1887-1898 Go to original source...
  11. DE CLERCQ P., PEETERS I., VERGAUWE G. & THAS O. 2003: Interaction between Podisus maculiventris and Harmonia axyridis, two predators used in augmentative biological control in greenhouse crops. - BioControl 48: 39-55 Go to original source...
  12. GAGNON A.E., DOYON J., HEIMPEL G.E. & BRODEUR J. 2011a: Prey DNA detection success following digestion by intraguild predators: Influence of prey and predator species. - Mol. Ecol. Resour. 11: 1022-1032 Go to original source...
  13. GAGNON A.E., HEIMPEL G.E. & BRODEUR J. 2011b: The ubiquity of intraguild predation among predatory arthropods. - Plos One 6(11): e28061. doi:10.1371/journal.pone.0028061 Go to original source...
  14. GARDINER M.M. & LANDIS D.A. 2007: Impact of intraguild predation by adult Harmonia axyridis (Coleoptera: Coccinellidae) on Aphis glycines (Hemiptera: Aphididae) biological control in cage studies. - Biol. Contr. 40: 386-395 Go to original source...
  15. GREENSTONE M.H., ROWLEY D.L., WEBER D.C., PAYTON M.E. & HAWTHORNE D.J. 2007: Feeding mode and prey detectability half-lives in molecular gut-content analysis: An example with two predators of the colorado potato beetle. - Bull. Entomol. Res. 97: 201-209 Go to original source...
  16. GREENSTONE M.H., SZENDREI Z., PAYTON M.E., ROWLEY D.L., COUDRON T.C. & WEBER D.C. 2010: Choosing natural enemies for conservation biological control: Use of the prey detectability half-life to rank key predators of colorado potato beetle. - Entomol. Exp. Appl. 136: 97-107 Go to original source...
  17. HAGLER J.R. & NARANJO S.E. 1997: Measuring the sensitivity of an indirect predator gut content elisa: Detectability of prey remains in relation to predator species, temperature, time, and meal size. - Biol. Contr. 9: 112-119 Go to original source...
  18. HALL T.A. 1999: Bioedit: A user-friendly biological sequence alignment editor and analysis program for windows 95/98/nt. - Nucl. Acids Symp. Ser. 41: 95-98
  19. HARPER G.L., KING R.A., DODD C.S., HARWOOD J.D., GLEN D.M., BRUFORD M.W. & SYMONDSON W.O.C. 2005: Rapid screening of invertebrate predators for multiple prey DNA targets. - Mol. Ecol. 14: 819-827 Go to original source...
  20. HARWOOD J.D. & OBRYCKI J.J. 2005: Quantifying aphid predation rates of generalist predators in the field. - Eur. J. Entomol. 102: 335-350 Go to original source...
  21. HAUTIER L., GREGOIRE J.C., DE SCHAUWERS J., SAN MARTIN Y GOMEZ G., CALLIER P., JANSEN J.P. & DE BISEAU J.C. 2008: Intraguild predation by Harmonia axyridis on coccinellids revealed by exogenous alkaloid sequestration. - Chemoecology 18: 191-196 Go to original source...
  22. HAUTIER L., SAN MARTIN Y GOMEZ G., CALLIER P., DE BISEAU J.C. & GREGOIRE J.C. 2011: Alkaloids provide evidence of intraguild predation on native coccinellids by Harmonia axyridis in the field. - Biol. Invasions 13: 1805-1814 Go to original source...
  23. HEMPTINNE J.L., MAGRO A., SALADIN C. & DIXON A.F.G. 2012: Role of intraguild predation in aphidophagous guilds. - J. Appl. Entomol. 136: 161-170 Go to original source...
  24. HODEK I. & HONEK A. 1996: Ecology of Coccinellidae. Kluwer, Dordrecht, Boston, London, 464 pp
  25. HOOGENDOORN M. & HEIMPEL G.E. 2001: PCR-based gut content analysis of insect predators: Using ribosomal its-1 fragments from prey to estimate predation frequency. - Mol. Ecol. 10: 2059-2067 Go to original source...
  26. HOSSEINI R., SCHMIDT O. & KELLER M.A. 2008: Factors affecting detectability of prey DNA in the gut contents of invertebrate predators: A polymerase chain reaction-based method. - Entomol. Exp. Appl. 126: 194-202 Go to original source...
  27. HOY M.A. 1994: Insect Molecular Genetics: An Introduction to Principals and Applications. Academic Press, San Diego, CA, 546 pp
  28. INGELS B. & DE CLERCQ P. 2011: Effect of size, extraguild prey and habitat complexity on intraguild interactions: A case study with the invasive ladybird Harmonia axyridis and the hoverfly Episyrphus balteatus. - BioControl 56: 871-882 Go to original source...
  29. JUEN A. & TRAUGOTT M. 2005: Detecting predation and scavenging by DNA gut-content analysis: A case study using a soil insect predator-prey system. - Oecologia 142: 344-352 Go to original source...
  30. KING R.A., READ D.S., TRAUGOTT M. & SYMONDSON W.O.C. 2008: Molecular analysis of predation: A review of best practice for DNA-based approaches. - Mol. Ecol. 17: 947-963 Go to original source...
  31. LUCAS E. 2005: Intraguild predation among aphidophagous predators. - Eur. J. Entomol. 102: 351-363 Go to original source...
  32. LUNDGREN J.G. & WEBER D.C. 2010: Changes in digestive rate of a predatory beetle over its larval stage: Implications for dietary breadth. - J. Insect Physiol. 56: 431-437 Go to original source...
  33. MCCORNACK B.P., KOCH R.L. & RAGSDALE D.W. 2007: A simple method for in-field sex determination of the multicolored asian lady beetle Harmonia axyridis. - J. Insect Sci. 7: 1-12 Go to original source...
  34. MCCULLAGH P. & NELDER J. 1989: Generalized Linear Models. Chapman and Hall, London, 551 pp
  35. MCEWEN P., NEW T.R. & WHITTINGTON A.E. 2001: Lacewings in the Crop Environment. Cambridge University Press, Cambridge, 546 pp
  36. MCMILLAN S., KUUSK A.K., CASSEL-LUNDHAGEN A. & EKBOM B. 2007: The influence of time and temperature on molecular gut content analysis: Adalia bipunctata fed with Rhopalosiphum padi. - Insect Sci. 14: 353-358 Go to original source...
  37. MICHAUD J.P. 2005: On the assessment of prey suitability in aphidophagous Coccinellidae. - Eur. J. Entomol. 102: 385-390 Go to original source...
  38. MICHAUD J.P. & HARWOOD J.D. 2012: Quantifying the impact of Coccinellids on their prey. In Hodek I., van Emden H.F. & Honek A. (eds): Ecology and Behaviour of the Ladybird Beetles. Blackwell, Oxford, pp. 465-487
  39. OSAWA N. & NISHIDA T. 1992: Seasonal variation in elytral color polymorphism in Harmonia axyridis (the ladybird beetle): The role of non-random mating. - Heredity 69: 297-307 Go to original source...
  40. PELL J.K., BAVERSTOCK J., ROY H.E., WARE R.L. & MAJERUS M.E.N. 2008: Intraguild predation involving Harmonia axyridis: A review of current knowledge and future perspectives. - Biocontrol 53: 147-168 Go to original source...
  41. POLIS G.A., MYERS C.A. & HOLT R.D. 1989: The ecology and evolution of intraguild predation: Potential competitors that eat each other. - Annu. Rev. Ecol. Syst. 20: 297-330 Go to original source...
  42. PUTRA N.S., YASUDA H. & SATO S. 2009: Oviposition preference of two hoverfly species in response to risk of intraguild predation. - Appl. Entomol. Zool. 44: 29-36 Go to original source...
  43. RICCI C. & PONTI L. 2005: Seasonal food of Ceratomegilla notata (Coleoptera: Coccinellidae) in mountain environments of northern Italian Alps. - Eur. J. Entomol. 102: 527-530 Go to original source...
  44. ROSENHEIM J.A., LIMBURG D.D. & COLFER R.G. 1999: Impact of generalist predators on a biological control agent, Chrysoperla carnea: Direct observations. - Ecol. Appl. 9: 409-417 Go to original source...
  45. ROY H.E., DE CLERCQ P., LANWSON HANDLEY L.J., POLAND R.L., SLOGGETT J.J. & WAJNBERG E. 2011: Alien arthropod predators and parasitoids: an ecological approach. - BioControl 56: 375-382 Go to original source...
  46. ROY H.E., ADRIAENS T., ISAAC N.J.B., KENIS M., ONKELINX T., SAN MARTIN G., BROWN P.M.J., HAUTIER L., POLAND R., ROY D.B., COMONT R., ESCHEN R., FROST R., ZINDEL R., VAN VLAENDEREN J., NEDVED O., RAVN H.P., GREGOIRE J.C., DE BISEAU J.C. & MAES D. 2012: Invasive alien predator causes rapid declines of native european ladybirds. - Divers. Distrib. 18: 717-725 Go to original source...
  47. ROZEN S. & SKALETSKY H. 2000: Primer3 on the www for general users and biologist programmers. In Krawetz S. & Misener S. (eds): Bioinformatics Methods and Protocols: Methods in Molecular Biology. Humana Press, Totowa, NJ, pp. 303-386
  48. SADEGHI H. & GILBERT F. 2000: Aphid suitability and its relationship to oviposition preference in predatory hoverflies. - J. Anim. Ecol. 69: 771-784 Go to original source...
  49. SHEPPARD S.K. & HARWOOD J.D. 2005: Advances in molecular ecology: Tracking trophic links through predator-prey foodwebs. - Funct. Ecol. 19: 751-762 Go to original source...
  50. SHEPPARD S.K., BELL J., SUNDERLAND K.D., FENLON J., SKERVIN D. & SYMONDSON W.O.C. 2005: Detection of secondary predation by PCR analyses of the gut contents of invertebrate generalist predators. - Mol. Ecol. 14: 4461-4468 Go to original source...
  51. SPSS 2010: Ibm Spss Statistics 19 Core System User's Guide. SPSS inc., Chicago
  52. STUBBS A.E. & FALK S.J. 2002: British Hoverflies: An Illustrated Identification Guide. British Entomological and Natural History Society, Reading, 469 pp
  53. SUNDERLAND K.D. 1988: Quantitative methods for detecting invertebrate predation occurring in the field. - Ann. Appl. Biol. 112: 201-224 Go to original source...
  54. SYMONDSON W.O.C. 2002: Molecular identification of prey in predator diets. - Mol. Ecol. 11: 627-641 Go to original source...
  55. THOMAS A.P., TROTMAN J., WHEATLEY A., AEBI A., ZINDEL R. & BROWN P.M.J. 2012: Predation of native coccinellids by the invasive alien Harmonia axyridis (Coleoptera: Coccinellidae): Detection in Britain by PCR-based gut analysis. - Insect Conserv. Diver. 6: 20-27 Go to original source...
  56. TRAUGOTT M., ZANGERL P., JUEN A., SCHALLHART N. & PFIFFNER L. 2006: Detecting key parasitoids of lepidopteran pests by multiplex PCR. - Biol. Contr. 39: 39-46 Go to original source...
  57. TRILTSCH H. 1999: Food remains in the guts of Coccinella septempunctata (Coleoptera: Coccinellidae) adults and larvae. - Eur. J. Entomol. 96: 355-364
  58. WARE R.L. & MAJERUS M.E.N. 2008: Intraguild predation of immature stages of british and japanese coccinellids by the invasive ladybird Harmonia axyridis. - BioControl 53: 169-188 Go to original source...
  59. WEBER D.C. & LUNDGREN J.G. 2009: Detection of predation using QPCR: Effect of prey quantity, elapsed time, chaser diet, and sample preservation on detectable quantity of prey DNA. - J. Insect Sci. 9: 41 Go to original source...
  60. WELLS P.M. 2011: Intraguild Predation by Harmonia axyridis: Effects on Native Enemies and Aphid Suppression. PhD Thesis, Darwin College, Cambridge University, 189 pp
  61. ZAIDI R.H., JAAL Z., HAWKES N.J., HEMINGWAY J. & SYMONDSON W.O.C. 1999: Can multiple-copy sequences of prey DNA be detected amongst the gut contents of invertebrate predators? - Mol. Ecol. 8: 2081-2087 Go to original source...
  62. ZHANG D.X. & HEWITT G.M. 1996: Assessment of the universality and utility of a set of conserved mitochondrial COI primers in insects. - Insect. Mol. Biol. 6: 143-150 Go to original source...