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Analysis of differential gene expression in the midgut of Ornithodoros moubata following infection with Borrelia duttonii (Ornitho-Mialomic)

Project


Project code: FLI-IMED-08-Ri-0616, 01KI1804
Contract period: 01.01.2019 - 31.12.2019
Budget: 90,309 Euro
Purpose of research: Experimental development
Keywords: Arthropods, zoonosis, animal disease

In recent years, infectious diseases whose pathogens are transmitted by arthropods have become increasingly important in the field of zoonoses research. Numerous research projects focused on the diversity of arthropods, their role as pathogen reservoir and their experimental vector competence. New approaches for tick control are dependent on defining molecular interactions between hosts, ticks and pathogens to allow for discovery of key molecules that could be tested in vaccines or new generation therapeutics for intervention of tick–pathogen cycles. This study aims to investigate transcripts in the midgut of Ornithodoros moubata in three different physiological states (naïv, blood-engorged & B. duttonii infected) underlying the processes of blood digestion, pathogen uptake and immunity against B. duttonii. The objective is to identify genes that are differentially expressed in the respective physiological state and thus represent key molecules.

The project aimed to characterize midgut transcriptomes of Ornithodoros moubata female ticks in three different physiological states (unfed, fed and Borrelia duttonii-infected), and to identify genes whose expression is differentially regulated in the respective physiological state and therefore play key roles in blood feeding and immunity. From a total of ~92 million reads from nine O. moubata midgut samples representing three biological treatments a de novo transcriptome was assembled. The filtered transcriptome comprises 80.905 sequences with an average length of 288.5 bp and a maximum length of 6208 bp. Comparisons of the libraries led to identification of several significantly differentially expressed transcripts. Between unfed and fed females, 326 differentially expressed transcripts were identified of which 184 transcripts could be annotated. Of these, 57 genes were upregulated and 127 were downregulated in fed females compared to unfed females. Comparison of expression patterns between non-infected and infected ticks revealed 160 differentially expressed transcripts of which 85 transcripts could be annotated. Of these, 44 genes were upregulated and 41 were downregulated in B. duttonii-infected females compared to non-infected females. Comparison of expression profiles between different experimental conditions show that the identified transcripts are highly specific as only nine transcripts were shared in all experimental conditions. In total, the most numerous transcripts were those with catalytic and binding activities and those involved in metabolic and cellular processes. Differentially expressed transcripts included some that are potentially be involved in blood digestion and immune response. However, as no reference genome is available for any tick species, functional annotation is challenging and has to be further elucidated by proteomic approaches. Mandy Schäfer, Cornelia Silaghi, Dirk Höper, Florian Pfaff (2020) Early transcriptional changes in the midgut of Ornithodoros moubata after feeding and infection with Borrelia duttonii. In prep.

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Subjects

Framework programme

BMEL - research cluster

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