RESUMO
A growing number of indigenous trypanosomes have been reported to naturally infect a variety of Australian wildlife with some species of Trypanosoma implicated in the population decline of critically endangered marsupials. However, the mode of transmission of Australian trypanosomes is unknown since their vectors remain unidentified. Here we aimed to fill this current knowledge gap about the occurrence and identity of indigenous trypanosomes in Australian invertebrates by conducting molecular screening for the presence of Trypanosoma spp. in native ticks collected from south-west Australia. A total of 231 ticks (148 collected from vegetation and 83 retrieved directly from 76 marsupial hosts) were screened for Trypanosoma using a High-Resolution Melt (HRM) qPCR assay. An overall Trypanosoma qPCR positivity of 37% (46/125) and 34% (26/76) was detected in questing ticks and host-collected (i.e., feeding) ticks, respectively. Of these, sequencing revealed 28% (35/125) of questing and 28% (21/76) of feeding ticks were infected with one or more of the five species of trypanosome previously reported in this region (T. copemani, T. noyesi, T. vegrandis, T. gilletti, Trypanosoma sp. ANU2). This work has confirmed that Australian ticks are capable of harbouring several species of indigenous trypanosome and likely serve as their vectors.
RESUMO
Trypanosomes are blood-borne parasites infecting a range of mammalian hosts worldwide. In Australia, an increasing number of novel Trypanosoma species have been identified from various wildlife hosts, some of which are critically endangered. Trypanosoma noyesi is a recently described species of biosecurity concern, due to a close relationship to the South American human pathogen, Trypanosoma cruzi. This genetic similarity increases the risk for introduction of T. cruzi via a local vector. Unfortunately, there is a lack of knowledge concerning the vectorial capacity of Australian invertebrates for native Trypanosoma species. Australian ixodid ticks (Ixodidae), which are widespread ectoparasites of mammalian wildlife, have received the most attention as likely candidates for trypanosome transmission and have been previously implicated as vectors. However, as all studies to date have focused on blood-fed ticks collected directly from infected mammalian hosts, the question of whether ticks maintain a trypanosome infection between blood meals is unknown. In this study, we investigated the presence of Trypanosoma within 148 Australian adult and nymph questing ticks of the species Amblyomma triguttatum, Ixodes australiensis, Ixodes myrmecobii and larvae Ixodes spp., collected from an endemic region of south-west Australia. Using a novel HRM-qPCR detection method that can discriminate between species of Trypanosoma based on primer melting temperature (Tm), we report the first molecular detection of Trypanosoma DNA in Australian questing ticks, with 6 ticks DNA positive for T. noyesi. Additionally, the presence of intact T. noyesi parasites within all (n = 3) smeared gut and gland contents of questing ticks was confirmed using a fluorescence in situ hybridisation (FISH) assay. Whilst this study was unable to determine the in situ tissue location of trypanosomes for the purpose of discerning a potential route of transmission, these combined molecular and FISH smear data indicate that trypanosomes can persist in ticks between blood meals and that ticks are possibly vectors in the transmission of T. noyesi between native wildlife. Transmission experiments are still required to evaluate the competency of Australian ticks as vectors for T. noyesi. Nevertheless, these novel findings warrant further investigation concerning potential life stages and the development of trypanosomes in both Australian, and other, tick species.