Comparison of DNA and Carbon and Nitrogen Stable Isotope-based Techniques for Identification of Prior Vertebrate Hosts of Ticks.

Identification of the vertebrate hosts upon which hematophagous arthropods feed provides key information for understanding the ecology and transmission of vector-borne diseases. Bloodmeal analysis of ticks presents unique challenges relative to other vectors, given the long interval between bloodmeal acquisition and host-seeking, during which DNA degradation occurs. This study evaluates DNA-based and stable isotope-based bloodmeal analysis methodologies for the lone star tick, Amblyomma americanum (Linneaus, 1758), in an experimental study with chicken as the known host. We subjected ticks of different ages and environmental rearing conditions to three DNA-based approaches and a stable isotopic analysis, which relies on the natural variation of nitrogen ((15)N/(14)N) and carbon ((13)C/(12)C) isotopes. While all three DNA-based approaches were successful in identifying the bloodmeal host of the engorged nymphs, only the probe-based RT-PCR was able to detect host DNA in aged ticks, the success of which was low and inconsistent across age and rearing treatments. In contrast, the stable isotope analysis showed utility in determining the host across all ages of ticks when isotopic values of ticks were compared with a panel of candidate vertebrate species. There was a positive shift in both δ(13)C and δ(15)N in adult A. americanum until 34 wk postnymphal bloodmeal. Through analyzing the isotopic signatures of eight potential vertebrate host species, we determined that the magnitude of this isotopic shift that occurred with tick age was minor compared with the heterogeneity in the δ(15)N and δ(13)C signatures among species. These results suggest that stable isotopes are a useful tool for understanding tick-host interactions.

Colour quantitation for chemical spot tests for a controlled substances presumptive test database.

Crime scene investigators (CSIs) often encounter unknown powders, capsules, tablets, and liquids at crime scenes, many of which are controlled substances. Because most drugs are white powders, however, visual determination of the chemical identity is difficult. Colourimetric tests are a well-established method of presumptive drug identification. Positive tests are often reported differently, however, because two analysts may perceive colour or record colourimetric results in different ways. In addition to perceiving colour differently, it is very common for there to be poor visibility conditions (e.g. rain, darkness) while performing these tests, further obscuring the results. In order to address these concerns and to create uniformity in the reporting of on-site colourimetric test results, this study has evaluated two of the state-of-the-art apps (ColorAssist® and Colorimeter®) for reporting the colour test results quantitatively in red-green-blue (RGB) format. The compiled library database of presumptive test results contains over 3300 data points including over 800 unique drug/test combinations. Variations observed between test replicates, from performing a test on different days, recording with a different device type (e.g. iPod Touch, iPhone models 4, 5c, 5s, or 6), and using different quantities of drug are discussed. Overall, the least variation in Euclidian norm was observed using ColorAssist® with the camera light (25.1±22.1) while the variation between replicates and data recorded using different devices was similar. The resulting library is uploaded to a smartphone application aimed to aid in identifying and interpreting suspected controlled substance evidence. Copyright © 2016 John Wiley & Sons, Ltd.