Development of a Molecular Assay to Detect Predation on Anopheles gambiae Complex Larval Stages

Erika Schielke Department of Ecology and Evolutionary Biology, Yale University, New Haven, Connecticut; Unite de Recherche 016, Institut de Recherche pour le Développement et Institut de Recherche en Sciences de la Santé, Bobo-Dioulasso, Burkina Faso; Department of Biology, Tor Vergata University, Rome, Italy; Centre National de Recherche et de Formation sur le Paludisme, Ouagadougou, Burkina Faso

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Carlo Costantini Department of Ecology and Evolutionary Biology, Yale University, New Haven, Connecticut; Unite de Recherche 016, Institut de Recherche pour le Développement et Institut de Recherche en Sciences de la Santé, Bobo-Dioulasso, Burkina Faso; Department of Biology, Tor Vergata University, Rome, Italy; Centre National de Recherche et de Formation sur le Paludisme, Ouagadougou, Burkina Faso

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Gianmaria Carchini Department of Ecology and Evolutionary Biology, Yale University, New Haven, Connecticut; Unite de Recherche 016, Institut de Recherche pour le Développement et Institut de Recherche en Sciences de la Santé, Bobo-Dioulasso, Burkina Faso; Department of Biology, Tor Vergata University, Rome, Italy; Centre National de Recherche et de Formation sur le Paludisme, Ouagadougou, Burkina Faso

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N’Falé Sagnon Department of Ecology and Evolutionary Biology, Yale University, New Haven, Connecticut; Unite de Recherche 016, Institut de Recherche pour le Développement et Institut de Recherche en Sciences de la Santé, Bobo-Dioulasso, Burkina Faso; Department of Biology, Tor Vergata University, Rome, Italy; Centre National de Recherche et de Formation sur le Paludisme, Ouagadougou, Burkina Faso

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Jeffrey Powell Department of Ecology and Evolutionary Biology, Yale University, New Haven, Connecticut; Unite de Recherche 016, Institut de Recherche pour le Développement et Institut de Recherche en Sciences de la Santé, Bobo-Dioulasso, Burkina Faso; Department of Biology, Tor Vergata University, Rome, Italy; Centre National de Recherche et de Formation sur le Paludisme, Ouagadougou, Burkina Faso

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Adalgisa Caccone Department of Ecology and Evolutionary Biology, Yale University, New Haven, Connecticut; Unite de Recherche 016, Institut de Recherche pour le Développement et Institut de Recherche en Sciences de la Santé, Bobo-Dioulasso, Burkina Faso; Department of Biology, Tor Vergata University, Rome, Italy; Centre National de Recherche et de Formation sur le Paludisme, Ouagadougou, Burkina Faso

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We developed a molecular assay to detect predation on Anopheles gambiae sensu lato (s.l.) mosquitoes. This intergenic spacer ribosomal DNA polymerase chain reaction assay and restriction enzyme analysis uses An. gambiae-specific primers to detect mosquito DNA in the DNA extracts from whole invertebrate predators, which enables identification of species (An. gambiae s.s. versus An. arabiensis) and molecular forms (M versus S in An. gambiae s.s.). We show that An. gambiae s.l. DNA can be detected after ingestion by members of the families Lestidae (order Odonata) after four hours, Libellulidae (order Odonata) after six hours, and Notonectidae (order Hemiptera) after 24 hours. This method is an improvement over previously published methods because of ease of execution and increased time of detection after ingestion.

Author Notes

  • 1

    Service MW, 1993. Mosquito Ecology: Field Sampling Methods. London: Applied Science Publishers Ltd.

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  • 2

    Service MW, 1971. Studies on sampling larval populations of the Anopheles gambiae complex. Bull World Health Organ 45 :169–180.

  • 3

    Service MW, 1973. Mortalities of the larvae of the Anopheles gambiae Giles complex and detection of predators by the precipitin test. Bull Entomol Res 62 :359–369.

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  • 4

    Service MW, 1977. Mortalities of the immature stages of Species B of the Anopheles gambiae complex in Kenya: comparison between rice field and temporary pools, identification of predators, and effects of insecticidal spraying. J Med Entomol 13 :535–545.

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  • 5

    Scott JA, Brogdon WG, Collins FH, 1993. Identification of single specimens of the Anopheles gambiae complex by polymerase chain reaction. Am J Trop Med Hyg 49 :520–529.

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  • 6

    Favia G, della Torre A, Bagayoko M, Lanfrancotti A, Sagnon NF, Touré YT, Coluzzi M, 1997. Molecular identification of sympatric chromosomal forms of Anopheles gambiae and further evidence of their reproductive isolation. Insect Mol Biol 6 :377–383.

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  • 7

    Paskewitz SM, Collins FH, 1990. Use of the polymerase chain reaction to identify mosquito species of the Anopheles gambiae complex. Med Vet Entomol 4 :367–373.

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  • 8

    Santolamazza F, Della Torre A, Caccone A, 2004. Short report: a new polymerase chain reaction–restriction fragment length polymorphism method to identify Anopheles arabiensis from Anopheles gambiae and its two molecular forms from degraded DNA templates or museum samples. Am J Trop Med Hyg 70 :604–606.

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  • 9

    Morales ME, Wesson DM, Sutherland IW, Impoinvil DE, Mbogo CM, Githure JI, Beier JC, 2003. Determination of Anopheles gambiae larval DNA in the gut of insectivorous dragonfly (Libellulidae) nymphs by polymerase chain reaction. J Am Mosq Control Assoc 19 :163–165.

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