Dataset Information

A simple non-invasive method to collect soft tick saliva reveals differences in Ornithodoros moubata saliva composition between ticks infected and uninfected with Borrelia duttonii spirochetes

ABSTRACT: We developed a new simple method to assess the composition of proteins in the Ornithodoros moubata saliva. To collect naturally expectorated saliva from the ticks, we employed an artificial membrane feeding technique using a simple, chemically defined diet and submitted the collected native saliva samples for liquid chromatography-mass spectrometry (LC-MS) analysis. These experiments were conducted with groups of uninfected ticks as well as with B. duttonii infected ticks. The ticks exhibited a fair feeding response to the tested diet, engorgement rates reaching between 60-100% of ticks per feeding chamber. LC-MS analysis identified a total of 17 and 15 proteins in saliva samples from the uninfected and infected ticks respectively. Importantly, the analysis was sensitive enough to detect up to 9 different proteins in the saliva containing diet upon which as few as 6 nymphal ticks fed. Some of the proteins identified are well known for their immunomodulatory activity in a vertebrate host, whereas others were structural or “housekeeping” proteins and their finding in the naturally expectorated saliva confirms that they can be secreted and having some roles at the tick-host interface. Most notably, some of the proteins that have long been suspected as important for vector-pathogen interactions of Borrelia spirochetes were detected only in the samples from infected ticks, suggesting that their expression was altered by the persistent colonization of the tick’s salivary glands by spirochetes. The simple method described herein is an important addition to the toolbox available to study the vector-host-pathogen interactions in the rapidly feeding soft ticks.

INSTRUMENT(S): timsTOF Pro

ORGANISM(S): Borrelia Ornithodoros Moubata

TISSUE(S): Saliva

SUBMITTER: Filip Dyčka

LAB HEAD: Ryan O. Rego

PROVIDER: PXD038824 | Pride | 2023-01-10

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	201024_saliva_201116_09h46m57s_txt.zip	Other
	201024_saliva_GSH_B.zip	Other
	201024_saliva_GSH_Ch_1.zip	Other
	201024_saliva_GSH_Ch_2.zip	Other
	201024_saliva_GSH_Ch_3.zip	Other

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Project description:In addressing R. microplus - A. marginale interactions, we propose and test three linked hypotheses. The first is that the tick gene response is organ specific: the midgut gene regulation is unique during feeding and during acquisition of A. marginale as compared to the salivary gland. This distinction is relevant as the two organs serve very different roles in the transmission biology of A. marginale with early survival and replication within the midgut epithelium, composed of highly phagocytic cells, required for initial colonization while a second round of replication in the salivary gland acini, composed of highly secretory cells, is required for transmission of an infectious dose in the saliva. Importantly, both the midgut epithelium and salivary glands have been identified as separate and distinct barriers for transmission of A. marginale and thus represent two potential sites where transmission could be blocked. The second hypothesis to be tested is that the salivary gland transcriptome is temporally dynamic. Initiation of tick attachment and feeding involves secretion of a virtual pharmacopeia including lytic enzymes, anticoagulants, and inhibitors of the mammalian innate immune and nocioceptive systems. Concomitantly, the acini provide an environment where A. marginale replicates >100 fold and are secreted into the saliva. Prior studies show that duration of feeding is a critical component of transmission efficiency, with increased efficiency positively correlated with time of tick feeding. The third hypothesis to be tested is that A. marginale colonization does not significantly modulate the tick midgut and salivary gland transcriptome. This hypothesis is based on observations by ourselves and others that tick infection does not impart a significant fitness cost on the vector. This is in contrast to other bacterial and protozoal pathogens that have dramatic effects on success of tick attachment, engorgement, and survival. A. marginale, similar to other tick-borne pathogens in the Family Anaplasmataceeae, is believed to have evolved from an arthropod-specific bacterium with relatively late adaptation to specific niches in mammalian hosts. Consequently, we predict that A. marginale is well adapted to its tick vector and utilizes the normal signaling pathways of the feeding tick with few, if any, effects on the midgut and salivary gland transcriptome. In this manuscript, we report the testing of these three hypotheses and present the results in context of the vector-pathogen-mammalian host interaction at the time of transmission. A Roche NimbleGen high-density gene expression microarray was custom designed based on the expressed sequence tag (EST) database, B. microplus Gene Index Version 2 (BmiGI V2) for R. microplus. The expression level of 14,447 R. microplus genes was analyzed from total RNA extracted from 10 different tick tissue samples; 30 arrays were included since triplicates of each different sample were analyzed as follow: unfed (midgut and salivary glands), blood feeding (2 days midgut and 2, 6 and 9 days salivary glands), A. marginale-infected blood feeding (2 days midgut and 2, 6 and 9 days salivary glands).

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Dataset Information

A simple non-invasive method to collect soft tick saliva reveals differences in Ornithodoros moubata saliva composition between ticks infected and uninfected with Borrelia duttonii spirochetes

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