Abstract:
The widespread geographical distribution of Rhipicephalus decoloratus in southern Africa and its ability to
transmit the pathogens causing redwater, gallsickness and spirochaetosis in cattle makes this hematophagous
ectoparasite of economic importance. In South Africa, the most commonly used chemical acaricides to control
tick populations are pyrethroids and amitraz. The current amitraz resistance mechanism described in R. microplus,
from South Africa and Australia, involves mutations in the octopamine receptor, but it is unlikely that
this will be the only contributing factor to mediate resistance. Therefore, in this study we aimed to gain insight
into the more complex mechanism(s) underlying amitraz resistance in R. decoloratus using RNA-sequencing.
Differentially expressed genes (DEGs) were identified when comparing amitraz susceptible and resistant ticks in
the presence of amitraz while fed on bovine hosts. The most significant DEGs were further analysed using several
annotation tools. The predicted annotations from these genes, as well as KEGG pathways potentially point towards
a relationship between the α-adrenergic-like octopamine receptor and ionotropic glutamate receptors in
establishing amitraz resistance. All genes with KEGG pathway annotations were further validated using RT-qPCR
across all life stages of the tick. In susceptible ticks, the proposed model is that in the presence of amitraz, there is
inhibition of Ca2+ entry into cells and subsequent membrane hyperpolarization which prevents the release of
neurotransmitters. In resistant ticks, we hypothesize that this is overcome by ionotropic glutamate receptors
(NMDA and AMPA) to enhance synaptic transmission and plasticity in the presence of neurosteroids. Activation
of NMDA receptors initiates long term potentiation (LTP) which may allow the ticks to respond more rapidly and
with less stimulus when exposed to amitraz in future. Overactivation of the NMDA receptor and excitotoxicity is
attenuated by the estrone, NAD+ and ATP hydrolysing enzymes. This proposed pathway paves the way to future
studies on understanding amitraz resistance and should be validated using in vivo activity assays (through the
use of inhibitors or antagonists) in combination with metabolome analyses.
