First records of Hyalomma rufipes and Ixodes neitzi (Acari: Ixodidae) found on large carnivores in South Africa

Ixodid ticks (Acari: Ixodidae) are important disease vectors for large carnivores, but the composition of the tick communities that parasitize carnivores is poorly understood. We collected ticks from leopards ( Panthera pardus ) and brown hyenas ( Hyaena brunnea ) in the Soutpansberg Mountains, South Africa, to determine which species feed on these carnivores. We identified a total of eight tick species belonging to six genera, and recorded Ixodes neitzi and Hyalomma rufipes on P. pardus for the first time.

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Introduction
Ticks are among the most important vectors of pathogens of wildlife worldwide (Pfäffle et al., 2013;Jongejan and Uilenberg, 2004). The impact of tick-borne diseases is of increasing concern to large carnivores globally, as populations are often threatened or significantly declining (Ripple et al., 2014). Disease can be an important cause of carnivore mortality (Murray et al. 1999), leading to population declines (Trinkel and Angelici, 2016) and extirpation (Cleaveland et al., 2007), with associated cascading effects on communities (Hollings et al., 2014). Parasite infection can also have substantial sub-lethal effects on hosts, including reducing foraging efficiency and competitive ability (Barber et al. 2000), and lowering reproductive success (Simmons and Zuk, 1992).
To date, very few studies of ticks have been conducted on large predators in Africa, and only a handful of individual leopards (Panthera pardus) and brown hyenas (Hyaena brunnea) have been sampled for ticks (Table A. 1, Theiler 1962in Boomker et al. 1997, I.G. Horak unpublished data in Boomker et al. 1997, Horak et al. 2010, mostly due to the difficulty of sampling these wild animals. This study adds observations to the current knowledge of tick species hosted by wild carnivores by cataloguing the tick species present on P. pardus and H. brunnea in the Soutpansberg Mountains, South Africa.

Methods
The study focussed on the Luvhondo Private Nature Reserve and neighbouring properties, which are located in the western Soutpansberg Mountain Range, Limpopo Province, South Africa (29°26'E, 23°01'S, altitude ca. 1,420 m), an area recognised as a biodiversity hotspot (Foord et al., 2002). The large carnivore guild resident in the area consists of P. pardus and H. brunnea. Although local population density estimates of P. pardus were previously high (Chase Grey et al., 2013), they appear to be declining (Williams et al., 2017) Ticks were opportunistically collected from animals by hand for up to 30 minutes, the maximum time possible before the animals started to wake up. All encountered life stages of ticks were collected (as reflected in Table 1). Ticks were preserved in vials containing 70% ethanol and stored at room temperature. Identification of the collected ticks was conducted by HH at the Onderstepoort Veterinary Institute, Pretoria, South Africa (references for identification: Apanaskevich and Horak, 2008;Apanaskevich et al., 2007;Clifford et al., 1977;Robinson, 1926;Theiler, 1961;Voltzit and Keirans, 2003;Walker and Olwage, 1987;Walker et al., 2000).

Results and discussion
Eight tick species belonging to six genera were identified from P. pardus (6 animals, 67 ticks) H. rufipes is a two-host tick, which spends its first two parasitic life stages, larva and nymph, on a single avian, leporid or rodent host and its adult stage on a large mammalian host (Dransfield and Brightfield, 2013). Since this tick species is not known to be associated with wild carnivores, and we collected only a single specimen from a single P. pardus, this may be an incidental finding. In contrast, I. neitzi was found on three out of six P. pardus individuals sampled. The species has been mentioned in scientific literature only a few times. The first account, and most other records (14 animals infested), were associated with mountain reedbuck (Redunca fulvorufula) , Clifford et al., 1977. Besides occasional collections from a klipspringer (Oreotragus oreotragus) (Rechav et al., 1978), a greater kudu (Tragelaphus strepsiceros), and an eland (Tragelaphus oryx) , our results represent the first account for large carnivores. Other than these incidental collections, however, host association and life cycle characteristics are unknown .
While adults have been described as most active from late spring to summer (Clifford et al., 1977), it is interesting to note that two of our three samples were collected in winter (June and July) with only one from summer (December).
We notice a much larger number of tick species on P. pardus (52, this study included) compared to H. brunnea (9) ( Table A1). This may in part reflect sampling efforts on the two species. However, there is no real evidence of more intense screening for ticks on P. pardus compared to H. brunnea in the literature, and, to our knowledge, there are no published reports on the number of P. pardus compared to the number of H. brunnea being caught.
Compared to H. brunnea, P. pardus inhabits a wider variety of habitat, and is, accordingly, exposed to a wider diversity of tick species Skinner and Smithers, 1990).
H. brunnea is associated with the dryer parts of the southern savanna (Skinner and Smithers, 1990), while many of the tick species collected from leopards (Table A1), especially the Ixodes species, occur in more humid habitats. Whether habitat use differences or hostand/or tick-specific characteristics are at the basis of this contrast remains to be studied.

Acknowledgements
The permit number for the capture of mammals was 001-CPM402-00003 and issued by the

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