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1 December 2013 Improving the Sterile Sperm Identification Method for Its Implementation in the Area-Wide Sterile Insect Technique Program Against Ceratitis capitata (Diptera: Tephritidae) in Spain
M. Juan-Blasco, A. Urbaneja, V. San AndréS, P. Castan˜ Era, B. Sabater-Mun˜ Oz
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Abstract

The success of sterile males in area-wide sterile insect technique (aw-SIT) programs against Ceratitis capitata (Wiedemann) is currently measured by using indirect methods as the wild:sterile male ratio captured in monitoring traps. In the past decade, molecular techniques have been used to improve these methods. The development of a polymerase chain reaction-restriction fragment-length polymorphism-based method to identify the transfer of sterile sperm to wild females, the target of SIT, was considered a significant step in this direction. This method relies on identification of sperm by detecting the presence of Y chromosomes in spermathecae DNA extract complemented by the identification of the genetic origin of this sperm: Vienna-8 males or wild haplotype. However, the application of this protocol to aw-SIT programs is limited by handling time and personnel cost. The objective of this work was to obtain a high-throughput protocol to facilitate the routine measurement in a pest population of sterile sperm presence in wild females. The polymerase chain reaction-restriction fragment-length polymorphism markers previously developed were validated in Mediterranean fruit fly samples collected from various locations worldwide. A laboratory protocol previously published was modified to allow for the analysis of more samples at the same time. Preservation methods and preservation times commonly used for Mediterranean fruit fly female samples were assessed for their influence on the correct molecular detection of sterile sperm. This high-throughput methodology, as well as the results of sample management presented here, provide a robust, efficient, fast, and economical sterile sperm identification method ready to be used in all Mediterranean fruit fly SIT programs.

© 2013 Entomological Society of America
M. Juan-Blasco, A. Urbaneja, V. San AndréS, P. Castan˜ Era, and B. Sabater-Mun˜ Oz "Improving the Sterile Sperm Identification Method for Its Implementation in the Area-Wide Sterile Insect Technique Program Against Ceratitis capitata (Diptera: Tephritidae) in Spain," Journal of Economic Entomology 106(6), 2541-2547, (1 December 2013). https://doi.org/10.1603/EC13064
Received: 1 February 2013; Accepted: 1 August 2013; Published: 1 December 2013
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KEYWORDS
mating assessment
Mediterranean fruit fly
PCR-restriction fragment-length polymorphism
stored sperm identification
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