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Tracking the sources of blood meals of parasitic arthropods using shotgun proteomics and unidentified tandem mass spectral libraries

Nature Protocols volume 9, pages 842–850 (2014)Cite this article

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Abstract

Identifying the species on which hematophagous arthropods feed is crucial for studying the factors that affect pathogen distributions and that can aid public health. Here we describe a protocol to identify the species a parasitic arthropod has previously fed upon by identifying the source of the remnants of a previous blood meal via shotgun proteomics and spectral matching. The protocol is a nontargeted approach that uses the entire detected blood proteome for source identification; it does not require a priori knowledge of genome or protein sequences. Instead, reference spectral libraries are compiled from the blood of multiple host species by using SpectraST, which takes ∼4 d; the identification of the species from which a previous blood meal of a hematophagous arthropod was taken is achieved with spectral matching against the reference spectral libraries, which takes approximately another 4 d. This method is robust against random degradation of the blood meal and can identify unknown blood remnants months after the feeding event.

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Figure 1: Data analysis workflow as described in Steps 15–18.
Figure 2: Screenshots of running msconvert (ProteoWizard) to convert vendor-specific formats to open XML data formats.
Figure 3: A snapshot illustrating the anticipated fingerprinting summary file.

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Acknowledgements

This work was supported by a grant from the US National Institutes of Health (AI076342); the US Center for Disease Control and Prevention (U01CK000170); and the University Grant Council, Hong Kong Special Administrative Region Government (HKUST RPC10EG08, DAG12EG01S).

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Authors and Affiliations

  1. Department of Biology, University of Pennsylvania, Philadelphia, Pennsylvania, USA

    Özlem Önder & Dustin Brisson

  2. Division of Biomedical Engineering, Bioengineering Graduate Program, The Hong Kong University of Science and Technology, Hong Kong, China

    Wenguang Shao & Henry Lam

  3. Department of Chemical and Biomolecular Engineering, The Hong Kong University of Science and Technology, Hong Kong, China

    Henry Lam

Authors
  1. Özlem Önder
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  2. Wenguang Shao
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  3. Henry Lam
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  4. Dustin Brisson
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Contributions

D.B., H.L. and Ö.Ö. conceived the study and formulated the experimental and computational strategy. Ö.Ö. optimized all the experimental protocols. H.L. and W.S. adapted SpectraST to build spectral libraries from unidentified spectra. Ö.Ö., H.L. and D.B. co-wrote the manuscript, with input from all authors.

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Correspondence to Özlem Önder or Henry Lam.

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Önder, Ö., Shao, W., Lam, H. et al. Tracking the sources of blood meals of parasitic arthropods using shotgun proteomics and unidentified tandem mass spectral libraries. Nat Protoc 9, 842–850 (2014). https://doi.org/10.1038/nprot.2014.048

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