ABSTRACT
Ticks, like other obligatory blood-feeding arthropods, rely on endosymbiotic bacteria to supplement their diet with B vitamins lacking in blood. It has been suggested that additional metabolites such as L-proline may be involved in this nutritional symbiosis, but this has yet to be tested. Here, we studied the metabolite-based interaction between the brown dog tick Rhipicephalus sanguineus (Acari: Ixodidae) and its Coxiella-like endosymbionts (CLE). We measured amino acid titers and tested the effect of B vitamins and L-proline supplementation on the fitness of CLE-suppressed female ticks, displaying low titers of CLE. We found higher titers of L-proline in the symbiont-hosting organs of unfed ticks and in engorged blood-fed whole ticks. Supplementation of B vitamins increased the hatching rate of CLE-suppressed ticks; this effect appears to be stronger when L-proline is added. Our results indicate that L-proline is produced by CLE, and we suggest that CLE is essential in states of high metabolic demand that affects tick reproductive fitness, such as oogenesis and embryonic development. These findings demonstrate the broader effect of nutritional symbionts on their hosts and may potentially contribute to the control of ticks and tick-borne diseases. IMPORTANCE: Coxiella-like endosymbionts (CLE) are essential to the brown dog tick Rhipicephalus sanguineus for feeding and reproduction. This symbiosis is based on the supplementation of B vitamins lacking in the blood diet. The involvement of additional metabolites has been suggested, but no experimental evidence is available as yet to confirm a metabolic interaction. Here, we show that B vitamins and L-proline, both of which contribute to tick reproductive fitness, are produced by CLE. These findings demonstrate the importance of symbiont-derived metabolites for the host's persistence and shed light on the complex bacteria-host metabolic interaction, which can be channeled to manipulate and control tick populations.
ABSTRACT
Obligatory hematophagous arthropods such as lice, bugs, flies, and ticks harbor bacterial endosymbionts that are expected to complement missing essential nutrients in their diet. Genomic and some experimental evidence support this expectation. Hard ticks (Acari: Ixodidae) are associated with several lineages of bacterial symbionts, and very few were experimentally shown to be essential to some aspects of tick's fitness. In order to pinpoint the nature of interactions between hard ticks and their symbionts, we tested the effect of massive elimination of Coxiella-like endosymbionts (CLE) by antibiotics on the development and fitness of the brown dog tick (Rhipicephalus sanguineus). Administration of ofloxacin to engorged (blood fed) nymphs resulted in significant and acute reduction of their CLE loads - an effect that also persisted in subsequent life stages (aposymbiotic ticks). As a result, the post-feeding development of aposymbiotic female (but not male) nymphs was delayed. Additionally, aposymbiotic adult females needed a significantly prolonged feeding period in order to replete (detach from host), and had reduced engorgement weight and a lower capacity to produce eggs. Consequently, their fecundity and fertility were significantly reduced. Eggs produced by aposymbiotic females were free of CLE, and the resulting aposymbiotic larvae were unable to feed successfully. Our findings demonstrate that the observed fitness effects are due to CLE reduction and not due to antibiotic administration. Additionally, we suggest that the contribution of CLE is not mandatory for oocyte development and embryogenesis, but is required during feeding in females, when blood meal processing and tissue buildup are taking place. Presumably, under these extreme physiological demands, CLE contribute to R. sanguineus through supplementing essential micro- and macronutrients. Further nutrient complementary studies are required to support this hypothesis.
