1 Larvae of Nuttalliella sp. (N. namaqua?) collected from small mammals in Limpopo Province, South Africa. http://ojvr.org/index.php/ojvr/article/downloadSuppFile/405/321
Because the larvae of N. namaqua have not previously been collected from natural hosts, nor been described, we relied on morphological features resembling those described for the females to arrive at a diagnosis. As an adjunct to identification of the larvae on taxonomic features, total genomic DNA from two larvae collected from M. namaquensis was extracted using the Qiagen, DNeasy® Blood and Tissue kit. Polymerase chain reaction and sequencing was performed on the 18S rRNA gene fragment using conserved universal primers. Amplifications were performed following standard PCR protocols and cycle-sequencing reactions were performed using BigDye Chemistry. Products were analysed on an automated sequencer (ABI 3730 XL DNA Analyzer, Applied Biosystems). A 466 base-pair region of the 18S rRNA gene was obtained for both larvae, and these sequences were aligned to the N. namaqua sequence on GenBank (Mans et al. 2011; JF751071.1). The region sequenced corresponds to positions 1093–1571 of the published sequence. The sequences generated for the two larvae in our study were identical. BLASTn searches on GenBank revealed 99.58% similarity (476/478) between the newly sampled individuals (GenBank Access Number JQ424828) and the 18S rRNA Nuttalliella sequence published on GenBank. Further support for the authenticity of our identification of the larvae was obtained by doing a GenBank BLASTn search against the entire database. The next closest alternative match to Nuttalliella would be the genus Ixodes, but in this instance the level of differentiation is more than 4% (614/640 identities between the larval sequences and that of Ixodes persulcatus: AY274888.1).Conclusion The larvae of a nuttalliellid tick, Nuttalliella sp. (N. namaqua?) can now be added to the immature stages of a large variety of ixodid tick species, which by preference feed on murid rodents. Acknowledgements We thank the management and staff of the Goro Game Reserve for permission to collect animals in the reserve and their continuous support. Stacey Hallam is thanked for collecting the ticks from mice in the Northern Cape Province. The sequences were generated by Lee-Gavin Williams and primers were provided by Anne Ropiquet. KM acknowledges a doctoral grant from the National Research Foundation (NRF), HL a Research Fellowship from the University of Pretoria and IGH funding from the University of Pretoria and the NRF. Competing interests The authors declare that they have no financial or personal relationship(s) which may have inappropriately influenced them in writing this paper. Authors’ contributions I.G.H. (University of Pretoria) identified the ticks with the assistance of D.A.A. (Georgia Southern University), and also compiled the manuscript. H.L. (University of Pretoria) and K.M. (University of Pretoria) collected the ticks and did the statistical work, and C.A.M. (Stellenbosch University) was responsible for the molecular analysis. 1.http://dx.doi.org/10.4102/ojvr.v78i1.2432.http://dx.doi.org/10.1371/journal pone.00236753.http://dx.doi.org/10.1645/GE-819R2.1174369414.http://dx.doi.org/10.3377/004.045.02225.PetneyT.NHorakI.GHowellD.JMeyerS2004Striped mice, Rhabdomys pumilio, and other murid rodents as hosts for immature ixodid ticks7131331815732458