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      Imaging of Borrelia turicatae Producing the Green Fluorescent Protein Reveals Persistent Colonization of the Ornithodoros turicata Midgut and Salivary Glands from Nymphal Acquisition through Transmission

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          Relapsing fever (RF) spirochetes colonize and are transmitted to mammals primarily by Ornithodoros ticks, and little is known regarding the pathogen's life cycle in the vector. To further understand vector colonization and transmission of RF spirochetes, Borrelia turicatae expressing a green fluorescent protein (GFP) marker ( B. turicatae-gfp) was generated. The transformants were evaluated during the tick-mammal infectious cycle, from the third nymphal instar to adult stage. B. turicatae-gfp remained viable for at least 18 months in starved fourth-stage nymphal ticks, and the studies indicated that spirochete populations persistently colonized the tick midgut and salivary glands. Our generation of B. turicatae-gfp also revealed that within the salivary glands, spirochetes are localized in the ducts and lumen of acini, and after tick feeding, the tissues remained populated with spirochetes. The B. turicatae-gfp generated in this study is an important tool to further understand and define the mechanisms of vector colonization and transmission.

          IMPORTANCE In order to interrupt the infectious cycle of tick-borne relapsing fever spirochetes, it is important to enhance our understanding of vector colonization and transmission. Toward this, we generated a strain of Borrelia turicatae that constitutively produced the green fluorescent protein, and we evaluated fluorescing spirochetes during the entire infectious cycle. We determined that the midgut and salivary glands of Ornithodoros turicata ticks maintain the pathogens throughout the vector's life cycle and remain colonized with the spirochetes for at least 18 months. We also determined that the tick's salivary glands were not depleted after a transmission blood feeding. These findings set the framework to further understand the mechanisms of midgut and salivary gland colonization.

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          Author and article information

          Role: Editor
          Appl Environ Microbiol
          Appl. Environ. Microbiol
          Applied and Environmental Microbiology
          American Society for Microbiology (1752 N St., N.W., Washington, DC )
          16 December 2016
          15 February 2017
          1 March 2017
          : 83
          : 5
          [a ]Department of Pediatrics, National School of Tropical Medicine, Baylor College of Medicine, Houston, Texas, USA
          [b ]Department of Biological Sciences, Mississippi State University, Starkville, Mississippi, USA
          [c ]U.S. Department of Agriculture-Agricultural Research Service, Knipling-Bushland U.S. Livestock Insects Research Laboratory and Veterinary Pest Genomics Center, Kerrville, Texas, USA
          [d ]Department of Entomology, Texas A&M AgriLife Research, Texas A&M University, College Station, Texas, USA
          [e ]Department of Molecular Virology and Microbiology, Baylor College of Medicine, Houston, Texas, USA
          University of Georgia
          Author notes
          Address correspondence to Job E. Lopez, job.lopez@ 123456bcm.edu .

          Present address: William K. Boyle, Creighton University School of Medicine, Omaha, Nebraska, USA.

          A.K. and H.K.W. contributed equally to this article.

          Citation Krishnavajhala A, Wilder HK, Boyle WK, Damania A, Thornton JA, Pérez de León AA, Teel PD, Lopez JE. 2017. Imaging of Borrelia turicatae producing the green fluorescent protein reveals persistent colonization of the Ornithodoros turicata midgut and salivary glands from nymphal acquisition through transmission. Appl Environ Microbiol 83:e02503-16. https://doi.org/10.1128/AEM.02503-16.

          PMC5311397 PMC5311397 5311397 02503-16
          Copyright © 2017 American Society for Microbiology.
          Page count
          supplementary-material: 3, Figures: 10, Tables: 2, Equations: 0, References: 27, Pages: 14, Words: 6952
          Funded by: HHS | NIH | NIH Office of the Director (OD) https://doi.org/10.13039/100000052
          Award ID: AI103724
          Award ID: AI091652
          Award Recipient : Job E. Lopez
          Funded by: HHS | NIH | NIH Office of the Director (OD) https://doi.org/10.13039/100000052
          Award ID: 1P20GM103646-01A1
          Award Recipient : Justin Adam Thornton
          Funded by: DOD | Defense Threat Reduction Agency (DTRA) https://doi.org/10.13039/100000774
          Award ID: U.S.C. 3318(b)-15217
          Award Recipient : Job E. Lopez
          Funded by: DOD | Defense Threat Reduction Agency (DTRA) https://doi.org/10.13039/100000774
          Award ID: U.S.C. 3318(b)-15217
          Award Recipient : Adalberto A. Perez de Leon
          Genetics and Molecular Biology
          Custom metadata
          March 2017


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