Evaluations of mosquito age grading techniques based on morphological changes

Evaluations were made of the accuracy and practicality of mosquito age grading methods based on changes to mosquito morphology; including the Detinova ovarian tracheation, midgut meconium, Polovodova ovariole dilatation, ovarian injection, and daily growth line methods. Laboratory maintained Aedes vigilax (Skuse) and Culex annulirostris (Skuse) females of known chronological and physiological ages were used for these assessments. Application of the Detinova technique to laboratory reared Ae. vigilax females in a blinded trial enabled the successful identification of nulliparous and parous females in 83.7-89.8% of specimens. The success rate for identifying nulliparous females increased to 87.8-98.0% when observations of ovarian tracheation were combined with observations of the presence of midgut meconium. However, application of the Polovodova method only enabled 57.5% of nulliparous, 1-parous, 2-parous, and 3-parous Ae. vigilax females to be correctly classified, and ovarian injections were found to be unfeasible. Poor correlation was observed between the number of growth lines per phragma and the calendar age of laboratory reared Ae. vigilax females. In summary, morphological age grading methods that offer simple two-category predictions (ovarian tracheation and midgut meconium methods) were found to provide high-accuracy classifications, whereas methods that offer the separation of multiple age categories (ovariolar dilatation and growth line methods) were found to be extremely difficult and of low accuracy. The usefulness of the morphology-based methods is discussed in view of the availability of new mosquito age grading techniques based on cuticular hydrocarbon and gene transcription changes.