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Background  Mosquito age and species identification is a crucial determinant of the efficacy of vector control programmes. Near-infrared spectroscopy (NIRS) has previously been applied successfully to rapidly, non-destructively, and simultaneously determine the age and species of freshly anesthetized African malaria vectors from the  Anopheles gambiae s.l . species complex:  An. gambiae s. s . and  Anopheles arabiensis . However, this has only been achieved on freshly-collected specimens and future applications will require samples to be preserved between field collections and scanning by NIRS. In this study, a sample preservation method (RNA later  ® ) was evaluated for mosquito age and species identification by NIRS against scans of fresh samples.    Methods  Two strains of  An. gambiae s.s . (CDC and G3) and two strains of  An. arabiensis  (Dongola, KGB) were reared in the laboratory while the third strain of  An. arabiensis  (Ifakara) was reared in a semi-field system. All mosquitoes were scanned when fresh and rescanned after preservation in RNA later  ®  for several weeks. Age and species identification was determined using a cross-validation.    Results  The mean accuracy obtained for predicting the age of young (<7 days) or old (≥ 7 days) of all fresh (n = 633) and all preserved (n = 691) mosquito samples using the cross-validation technique was 83% and 90%, respectively. For species identification, accuracies were 82% for fresh against 80% for RNA later  ®  preserved. For both analyses, preserving mosquitoes in RNA later  ®  was associated with a highly significant reduction in the likelihood of a misclassification of mosquitoes as young or old using NIRS. Important to note is that the costs for preserving mosquito specimens with RNA later  ®  ranges from 3-13 cents per insect depending on the size of the tube used and the number of specimens pooled in one tube.    Conclusion  RNA later  ®  can be used to preserve mosquitoes for subsequent scanning and analysis by NIRS to determine their age and species with minimal costs and with accuracy similar to that achieved from fresh insects. Cold storage availability allows samples to be stored longer than a week after field collection. Further study to develop robust calibrations applicable to other strains from diverse ecological settings is recommended.

Evaluating RNAlater® as a preservative for using near-infrared spectroscopy to predict Anopheles gambiae age and species