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Background  Vertebrate bloodfeeding is a critical component of a mosquito's ability to transmit pathogens that cause diseases such as malaria, dengue fever and viral encephalitis. Due to degradation by the digestive process, current methods to identify mosquito bloodmeal sources are only useful for approximately 36 hours post-feeding. A critical need exists for technologies to extend this window and gain a more complete picture of mosquito feeding behavior for epidemiological studies. Stable isotopes are useful for investigating organism feeding behavior because the isotopic ratio of an organism's tissues reflects that of the material it ingests.    Methodology/Principal Findings  Proof-of-principle data indicates that after bloodfeeding,  Aedes albopictus  mosquitoes acquire diagnostic Carbon and Nitrogen stable isotope profiles from their vertebrate hosts that can be accurately identified one week post-feeding, approximately 4 days after the entire bloodmeal has been digested. Total C/N ratio served as a biomarker marker for bloodfeeding ( P <0.02), while δN was the most informative variable which could distinguish between unfed, chicken-fed and human-fed mosquitoes ( P <0.01). By plotting C/N vs. δN, all feeding treatments could be identified in a double-blind analysis.    Conclusions/Significance  These proof-of-principle experiments indicate that analysis of stable isotopes can be used to distinguish bloodfed from unfed mosquitoes, and also distinguish between different vertebrate bloodmeal sources even after all blood has been digested. The development of stable isotope-based assays for mosquito bloodmeal identification may be a powerful tool to investigate mosquito feeding ecology and the dynamics of vector-borne pathogens.

Stable Isotope Analysis Can Potentially Identify Completely-Digested Bloodmeals in Mosquitoes