Exploring marking methods for the predatory hoverfly Sphaerophoria rueppellii (Diptera: Syrphidae)

Abstract

Hoverflies are essential to ecosystems, with adults serving as important pollinators and larvae preying on plant-feeding insects or recycling nutrients. Species like Sphaerophoria rueppellii are used in biocontrol programs to target aphid pests. To enhance these programs, markers can be used in a mark-release-recapture (MRR) method to track hoverfly feeding and oviposition sites. Effective markers must be persistent and not harm the hoverflies’ vital functions. This study evaluated three marking methods for S. rueppellii: rubidium (RbCl), fluorescein, and fluorescent dust. Laboratory experiments assessed the effects of these markers on female hoverfly fecundity, mating behavior and marking persistence. Results showed no significant differences in egg-laying or survival time between marked and unmarked females. Rubidium and fluorescein did not affect mating behavior, but dust-treated females mated significantly less than untreated females. In terms of marking persistence, rubidium and fluorescent dusts remained detectable throughout the hoverflies’ adult lifespan, while fluorescein markings faded within 24 h. Fluorescent dusts were easy-to-use, durable, and cost-effective, but careful application and further study are needed to avoid potential effects on insect activity and mating ability. Fluorescein showed no adverse effects on insect biology, was economical and quick to apply, but had short persistence, making it unsuitable for long-term field studies. Rubidium was harmless to insects and detectable for long periods, but its detection required financial investment, time, and specialized equipment. This research provides valuable insights into the potential of hoverflies as biocontrol agents and offers new tools for their effective management in agricultural settings.

Keywords

biocontrol / fluorescein / fluorescent dusts / hoverflies / insect tagging / rubidium

References

[1] Alcock, J. (1981) Lek territoriality in the Tarantula Hawk Wasp Hemipepsis ustulata (Hymenoptera: Pompilidae). Behavioral Ecology and Sociobiology, 8, 309–317.

[2] Alcock, J. (1987) Leks and hilltopping in insects. Journal of Natural History, 21, 319–328.

[3] Alcock, J. (2011) Hilltopping behavior by males of Tachysphex menkei Pulawski (Hymenoptera, Crabronidae). Journal of Hymenoptera Research, 21, 65–70.

[4] Alverson, D.R., All, J.N. and Bush, P.B. (1980) Rubidium as a marker and simulated inoculum for the black-faced leafhopper, Graminella nigrifrons, the primary vector of maize chlorotic dwarf virus of corn. Environmental Entomology, 9, 29–31.

[5] Amorós-Jiménez, R. and Marcos-García, M. (2020) Fitness-related parameters of the aphid predator Sphaerophoria rueppellii (Diptera, Syrphidae) feeding on three different aphid pests. Boletín de la Asociación Española de Entomología, 44, 299–315.

[6] Amorós-Jiménez, R., Pineda, A., Fereres, A. and Marcos-García, M.Á. (2012) Prey availability and abiotic requirements of immature stages of the aphid predator Sphaerophoria rueppellii. Biological Control, 63, 17–24.

[7] An, L., Neimann, A., Eberling, E., Algora, H., Brings, S. and Lunau, K. (2018) The yellow specialist: Dronefly Eristalis tenax prefers different yellow colours for landing and proboscis extension. Journal of Experimental Biology, 221, jeb184788.

[8] Argauer, R.J. and Cantelo, W.W. (1972) Spectrofluorometric determination of fluorescein-tagged Tobacco Hornworms. Journal of Economic Entomology, 65, 539–542.

[9] Aubert, J. and Goeldlin de Tiefenau, P. (1981) Observations on the migration of syrphids (Dipt.) in the Alps of Western Switzerland. Mineilungen Der Schweizerischen Entomologischen Gesellschaft, 54, 377–388.

[10] Ball, S.G. and Morris, R.K.A. (2004) A mark-release-recapture study of Volucella bombylans (Linnaeus), V. inflata (Fabricius) and V. pellucens (Linnaeus) (Diptera, Syrphidae). Dipterists Digest, 10, 73–83.

[11] Barlow, C.A. (1961) On the biology and reproductive capacity of Syrphus corollae Fab. (Syrphidae) in the laboratory. Entomologia Experimentalis et Applicata, 4, 91–100.

[12] Bellefeuille, Y., Fournier, M. and Lucas, E. (2021) Biological control of the foxglove aphid using a banker plant with Eupeodes americanus (Diptera: Syrphidae) in experimental and commercial greenhouses. Biological Control, 155, 104541.

[13] Benestad, E. (1970) Laboratory experiments on the biology of Syrphus corollae (Fabr.) (Dipt., Syrphidae). Norsk Entomologisk Tidskrift, 17, 77–85.

[14] Bishop, L.G. (1974) An ultraviolet photoreceptor in a Dipteran compound eye. Journal of Comparative Physiology, 91, 267–275.

[15] Bribosia, E., Bylemans, D., Migon, M. and van Impe, G. (2005) Internal labelling of the eggs of the aphidophagous hoverfly Episyrphus balteatus (De Geer) (Diptera: Syrphidae) with a fluorescent dye. Communications in Agricultural and Applied Biological Sciences, 70, 725–731.

[16] Briggs, E.L., Baranski, C., Münzer Schaetz, O., Garrison, G., Collazo, J.A. and Youngsteadt, E. (2022) Estimating bee abundance: Can mark-recapture methods validate common sampling protocols? Apidologie, 53, 10.

[17] Burgio, G., Dindo, M.L., Pape, T., Whitmore, D. and Sommaggio, D. (2024) Diptera as predators in biological control: applications and future perspectives. BioControl, 70, 1–17.

[18] Chapman, J.A. (1954) Studies on summit-frequenting insects in western Montana. Ecology, 35, 41–49.

[19] Clem, C.S., Sparbanie, T.M., Luro, A.B. and Harmon-Threatt, A.N. (2020) Can anthophilous hover flies (Diptera: Syrphidae) discriminate neonicotinoid insecticides in sucrose solution? PLoS ONE, 15, e0234820.

[20] Clymans, R., Van Kerckvoorde, V., Beliën, T., Bylemans, D. and De Clercq, P. (2020) Marking Drosophila suzukii (Diptera: Drosophilidae) with fluorescent dusts. Insects, 11, 152.

[21] Collett, T.S. and Land, M.F. (1975a) Visual control of flight behaviour in the hoverfly Syritta pipiens L. Journal of Comparative Physiology, 99, 1–66.

[22] Collett, T.S. and Land, M.F. (1975b) Visual spatial memory in a hoverfly. Journal of Comparative Physiology, 100, 59–84.

[23] Conn, D.L.T. (1976) Estimates of population size and longevity of adult narcissus bulb fly Merodon equestris Fab. (Diptera: Syrphidae). Journal of Applied Ecology, 13, 429–434.

[24] Coppedge, J.R., Spencer, J.P., Brown, H.E., Whitten, C.J., Snow, J.W. and Wright, J.E. (1979) A new dye marking technique for the screwworm. Journal of Economic Entomology, 72, 40–42.

[25] Corbett, A., Murphy, B.C., Rosenheim, J.A. and Bruins, P.F. (1996) Labeling an egg parasitoid, Anagrus epos (Hymenoptera: Mymaridae), with rubidium within an overwintering refuge. Environmental Entomology, 25, 29–38.

[26] Crumpacker, D.W. (1974) The use of micronized fluorescent dusts to mark adult Drosophila pseudoobscura. The American Midland Naturalist, 91, 118–129.

[27] Culbert, N.J., Kaiser, M., Venter, N., Vreysen, M.J.B., Gilles, J.R.L. and Bouyer, J. (2020) A standardised method of marking male mosquitoes with fluorescent dust. Parasites & Vectors, 13, 192.

[28] Dickens, B.L. and Brant, H.L. (2014) Effects of marking methods and fluorescent dusts on Aedes aegypti survival. Parasites & Vectors, 7, 65.

[29] Dinkel, T. and Lunau, K. (2001) How drone flies (Eristalis tenax L., Syrphidae, Diptera) use floral guides to locate food sources. Journal of Insect Physiology, 47, 1111–1118.

[30] Dominiak, B.C., Sundaralingam, S., Jiang, L., Jessup, A.J. and Barchia, I.M. (2010) Impact of marker dye on adult eclosion and flight ability of mass produced Queensland fruit fly Bactrocera tryoni (Froggatt) (Diptera: Tephritidae). Australian Journal of Entomology, 49, 166–169.

[31] Downes, J.A. (1969) The swarming and mating flight of diptera. Annual Review of Entomology, 14, 271–298.

[32] Fauteux, A., Soares, A.O. and Lucas, E. (2024) Larval development and voracity of Eupeodes americanus (Diptera: Syrphidae): comparison of the focal prey Aphis gossypii (Hemiptera: Aphididae) and the banker prey Rhopalosiphum padi (Hemiptera: Aphididae). Insect Science, 31, 575–586.

[33] Fitzpatrick, S. and Wellington, W. (1983) Contrasts in the territorial behavior of three species of hover flies (Diptera: Syrphidae). The Canadian Entomologist, 115, 559–566.

[34] Gilbert, F.S. (1984) Thermoregulation and the structure of swarms in Syrphus ribesii (Syrphidae). Oikos, 42, 249–255.

[35] Gilbert, F.S. and Falk, S.J. (2015) Hoverflies. Pelagic Publishing, Exeter, UK.

[36] Gonzalez, N., Buitenhuis, R. and Lucas, E. (2024) Comparison of two release strategies for the American hoverfly, Eupeodes americanus, against the green peach aphid in greenhouses: banker plant vs. pupal release. Crop Protection, 184, 106814.

[37] Graham, H.M. and Wolfenbarger, D.A. (1977) Tobacco budworm: labeling with rubidium in the laboratory. Journal of Economic Entomology, 70, 800–802.

[38] Grimm, A., Gruber, B. and Henle, K. (2014) Reliability of different mark-recapture methods for population size estimation tested against reference population sizes constructed from field data. PLoS ONE, 9, e98840.

[39] Haenke, S., Scheid, B., Schaefer, M., Tscharntke, T. and Thies, C. (2009) Increasing syrphid fly diversity and density in sown flower strips within simple vs. complex landscapes. Journal of Applied Ecology, 46, 1106–1114.

[40] Hagler, J.R. and Jackson, C.G. (2001) Methods for marking insects: current techniques and future prospects. Annual Review of Entomology, 46, 511–543.

[41] Holloway, G. and McCaffery, A.R. (1990) Habitat utilisation and dispersion in Eristalis pertinax (Diptera, Syrphidae). The Entomologist, 109, 116–124.

[42] Jönsson, A.M., Ekroos, J., Dänhardt, J., Andersson, G.K.S., Olsson, O. and Smith, H.G. (2015) Sown flower strips in southern Sweden increase abundances of wild bees and hoverflies in the wider landscape. Biological Conservation, 184, 51–58.

[43] Jost, D.J. and Pitre, H.N. (2002) Activity of rubidium and cesium in soybean looper (Lepidoptera: Noctuidae): insect feeding on cotton and soybean measured by elemental markers. Journal of Economic Entomology, 95, 278–285.

[44] Khelifa, R., Mahdjoub, H., M'Gonigle, L. and Kremen, C. (2021) Integrating high-speed videos in capture-mark-recapture studies of insects. Ecology and Evolution, 11, 6033–6040.

[45] Kirkpatrick, D.M., Rice, K.B., Ibrahim, A., Fleischer, S.J., Tooker, J.F., Tabb, A.et al. (2020) The influence of marking methods on mobility, survivorship, and field recovery of Halyomorpha halys (Hemiptera: Pentatomidae) adults and nymphs. Environmental Entomology, 49, 1026–1031.

[46] Klick, J., Yang, W.Q. and Bruck, D.J. (2015) Marking Drosophila suzukii (Diptera: Drosophilidae) with rubidium or ¹⁵N. Journal of Economic Entomology, 108, 1447–1451.

[47] Knight, A.L., Hull, L.A., Rajotte, E.G. and Fleischer, S.J. (1989) Labeling tufted apple bud moth (Lepidoptera: Tortricidae) with rubidium: effect on development, longevity, and fecundity. Annals of the Entomological Society of America, 82, 481–485.

[48] Leman, A., Mouratidis, A., Pijnakker, J., Vervoorn, K., Wäckers, F. and Messelink, G.J. (2023) Sugar and pollen supply enhances aphid control by hoverflies in strawberry. Biological Control, 186, 105347.

[49] Li, H., Wyckhuys, K.A.G. and Wu, K.M. (2023) Hoverflies provide pollination and biological pest control in greenhouse-grown horticultural crops. Frontiers in Plant Science, 14, 1118388.

[50] Lövei, G.L., MacLeod, A. and Hickman, J.M. (1998) Dispersal and effects of barriers on the movement of the New Zealand hover fly Melanostoma fasciatum (Dipt., Syrphidae) on cultivated land. Journal of Applied Entomology, 122, 115–120.

[51] Maciel-de-Freitas, R., Gonçalves, J. and Lourenço-de-Oliveira, R. (2004) Efficiency of rubidium marking in Aedes albopictus (Diptera: Culicidae): Preliminary evaluation on persistence of egg labeling, survival, and fecundity of marked female. Memórias do Instituto Oswaldo Cruz, 99, 823–827.

[52] MacKinnon, W., Eveleigh, E., Silk, P. and Forbes, G. (2016) Labeling feral spruce budworm (Lepidoptera: Tortricidae) populations with rubidium. Environmental Entomology, 45, 427–435.

[53] MacLeod, A. (1999) Attraction and retention of Episyrphus balteatus DeGeer (Diptera: Syrphidae) at an arable field margin with rich and poor floral resources. Agriculture, Ecosystems & Environment, 73, 237–244.

[54] Madeira, F. and Pons, X. (2016) Rubidium marking reveals different patterns of movement in four ground beetle species (Col., Carabidae) between adjacent alfalfa and maize. Agricultural and Forest Entomology, 18, 99–107.

[55] Maier, C.T. (1978) Immature stages and biology of Mallota posticata (Fabricius) (Diptera: Syrphidae). Proceedings of the Entomological Society of Washington, 80, 424–440.

[56] Maier, C.T. and Waldbauer, G.P. (1979) Dual mate-seeking strategies in male Syrphid Flies (Diptera: Syrphidae). Annals of the Entomological Society of America, 72, 54–61.

[57] Makumbe, L.D.M., Manrakhan, A. and Weldon, C.W. (2017) Marking Bactrocera dorsalis (Diptera: Tephritidae) with fluorescent pigments: effects of pigment colour and concentration. African Entomology, 25, 220–234.

[58] Messing, R.H., Klungness, L.M., Purcell, M. and Wong, T.T.Y. (1993) Quality control parameters of mass-reared opiine parasitoids used in augmentative biological control of tephritid fruit flies in Hawaii. Biological Control, 3, 140–147.

[59] Moffitt, H.R. and Albano, D.J. (1972) Codling moths: Fluorescent powders as markers. Environmental Entomology, 1, 750–753.

[60] Nakata, T. (2008) Effectiveness of micronized fluorescent powder for marking citrus psyllid. Diaphorina citri. Applied Entomology and Zoology, 43, 33–36.

[61] Nielsen, T. (1969) Population studies of Helophilus hybridus Loew and Sericomyia silentis (Harris) (Dipt., Syrphidae) on Jaeren SW Norway. Norsk Entomologisk Tidsskrift, 16, 33–38.

[62] Nixon, L.J., Ludwick, D.C. and Leskey, T.C. (2021) Horizontal and vertical dispersal capacity and effects of fluorescent marking on Lycorma delicatula nymphs and adults. Entomologia Experimentalis et Applicata, 169, 219–226.

[63] Odermatt, J., Frommen, J.G. and Menz, M.H.M. (2017) Consistent behavioural differences between migratory and resident hoverflies. Animal Behaviour, 127, 187–195.

[64] Orengo-Green, J.J., Casas, J.L. and Marcos-García, M.Á. (2022) Effect of abiotic climatic factors on the gonadal maturation of the biocontrol agent Sphaerophoria rueppellii (Wiedemann, 1830) (Diptera: Syrphidae). Insects, 13, 573.

[65] Ouattara, T.Y., Fournier, M., Gonzalez, N., Rojo, S., Lucas, E. (2024) Reproductive parameters of a new biocontrol agent, Eupeodes americanus (Diptera: Syrphidae) and comparison with the commercialized Aphidoletes aphidimyza (Diptera: Cecidomyiidae). Journal of Economic Entomology, 117, 1760–1768.

[66] Pekas, A., De Craecker, I., Boonen, S., Wäckers, F.L. and Moerkens, R. (2020) One stone; two birds: concurrent pest control and pollination services provided by aphidophagous hoverflies. Biological Control, 149, 104328.

[67] Pineda, A. and Marcos-Garcia, M.Á. (2008a) Introducing barley as aphid reservoir in sweet-pepper greenhouses: effects on native and released hoverflies (Diptera: Syrphidae). European Journal of Entomology, 105, 531–535.

[68] Pineda, A. and Marcos-García, M.Á. (2008b) Use of selected flowering plants in greenhouses to enhance aphidophagous hoverfly populations (Diptera: Syrphidae). Annales de la Société Entomologique de France, 44, 487–492.

[69] Pineda, A. and Marcos-García, M.A. (2008c) Evaluation of several strategies to increase the residence time of Episyrphus balteatus (Diptera, Syrphidae) releases in sweet pepper greenhouses. Annals of Applied Biology, 152, 271–276.

[70] Polavarapu, S., Lonergan, G.C. and Seabrook, W.D. (1992) Labelling the blueberry leaftier, Croesia curvalana with foliar sprays of rubidium chloride. Entomologia Experimentalis et Applicata, 63, 143–153.

[71] Prasifka, J.R., Heinz, K.M. and Sansone, C.G. (2001) Field testing rubidium marking for quantifying intercrop movement of predatory arthropods. Environmental Entomology, 30, 711–719.

[72] Roslin, T. (2000) Dung beetle movements at two spatial scales. Oikos, 91, 323–335.

[73] Rotheray, E., Bussiere, L., Moore, P., Bergstrom, L. and Goulson, D. (2014) Mark recapture estimates of dispersal ability and observations on the territorial behaviour of the rare hoverfly, Hammerschmidtia ferruginea (Diptera, Syrphidae). Journal of Insect Conservation, 18, 179–188.

[74] Rodríguez-Gasol, N., Alins, G., Veronesi, E.R. and Wratten, S. (2020) The ecology of predatory hoverflies as ecosystem-service providers in agricultural systems. Biological Control, 151, 104405.

[75] Rojas-Araya, D., Alto, B.W., Burkett-Cadena, N. and Cummings, D.A. (2020) Detection of fluorescent powders and their effect on survival and recapture of Aedes aegypti (Diptera: Culicidae). Journal of Medical Entomology, 57, 266–272.

[76] Rojo, S., Gilbert, F.S., Marcos-García, M., Nieto, J.M. and Mier, M.P. (2003) A World Review of Predatory Hoverflies (Diptera, Syrphidae: Syrphinae) and Their Prey. Centro Iberoamericano De La Biodiversidad (CIBIO), Murcia, Spain, pp. 319.

[77] Rotheray, G. and Gilbert, F. (1989) The phylogeny and systematics of European predacious Syrphidae (Diptera) based on larval and puparial stages. Zoological Journal of the Linnean Society, 95, 29–70.

[78] Sarkar, D., Muthukrishnan, S. and Sarkar, M. (2017) Fluorescent marked mosquito offer a method for tracking and study mosquito behaviour. International Journal of Mosquito Research, 4, 5–9.

[79] Schtickzelle, N., Le Boulengé, E. and Baguette, M. (2002) Metapopulation dynamics of the bog fritillary butterfly: Demographic processes in a patchy population. Oikos, 97, 349–360.

[80] Stimmann, M.W. (1974) Marking insects with rubidium: Imported cabbageworm marked in the field. Environmental Entomology, 3, 327–328.

[81] Tschumi, M., Albrecht, M., Collatz, J., Dubsky, V., Entling, M.H., Najar-Rodriguez, A.J.et al. (2016) Tailored flower strips promote natural enemy biodiversity and pest control in potato crops. Journal of Applied Ecology, 53, 1169–1176.

[82] Van Steenwyk, R.A., Ballmer, G.R., Page, A.L. and Reynolds, H.T. (1978) Marking pink bollworm with rubidium. Annals of the Entomological Society of America, 71, 81–84.

[83] Van Steenwyk, R.A., Kaneshiro, K.Y., Hue, N.V. and Whittier, T.S. (1992) Rubidium as an internal physiological marker for Mediterranean fruit fly (Diptera: Tephritidae). Journal of Economic Entomology, 85, 2357–2364.

[84] Verhulst, N.O., Loonen, J.A. and Takken, W. (2013) Advances in methods for colour marking of mosquitoes. Parasites & Vectors, 6, 200.

[85] Waldbauer, G. (1990) Hilltopping by males of Eupeodes Volucris (Diptera: Syrphidae). The Great Lakes Entomologist, 23, 175–176.

[86] Wolton, R.J. (2012) Observations on adult behaviour of Microdon myrmicae Schönrogge et al. (Diptera, Syrphidae), with particular reference to female survival, dispersal and oviposition. Dipterists Digest, 19, 103–106.

[87] Wratten, S.D., Bowie, M.H., Hickman, J.M., Evans, A.M., Sedcole, J.R. and Tylianakis, J.M. (2003) Field boundaries as barriers to movement of hover flies (Diptera: Syrphidae) in cultivated land. Oecologia, 134, 605–611.

[88] Zheng, Z., Liu, H., Wang, X., Wu, X., Chen, Y., Deng, J.et al. (2019) Development and reproduction of the hoverfly Eupeodes corollae (Diptera: Syrphidae). SDRP Journal of Earth Sciences & Environmental Studies, 4, 4.

Rights & permissions

2025 The Author(s). Insect Science published by John Wiley & Sons Australia, Ltd on behalf of Institute of Zoology, Chinese Academy of Sciences.