These questions include the study of Wolbachia population genetics within infected species [30, 38, 39], and will further extend studies of horizontal transmission between host species for which MLST was originally developed [22]. Highly polymorphic markers will also be useful for experimental evolution of Wolbachia in order to track small genomic changes in short time frames. This

higher resolution comes with the cost though, that markers are not universally applicable to the entire diversity of Wolbachia. (2) The majority of Wolbachia genomes are dotted with many different repeat regions which are highly appropriate to be targeted for the isolation of possible polymorphic markers. Tandem repeat markers such as the ones developed here can be tailored to individual studies. (3) MLVA markers are ideal for rapid Epigenetics Compound Library manufacturer and high-throughput DNA fingerprinting, as no sequencing is required. The markers are ideal to detect multiple infections in single PCR reactions if strains contain alleles with variable amplicon sizes. Our analysis of the evolution of the tandem repeat regions shows that they evolve by gain or loss of repeats. The variability in the number

of ANK repeats, generally constituted by 33 amino acids each, creates size differences that are multiples of 99bp and, like VNTRs consisting of >100bp periods, can be clearly identified following simple PCR screenings without the need of initial sequencing or RFLP analyses as in the case of point mutations. The use of 2-3 highly variable markers per strain can generate Poziotinib molecular weight easily readable fingerprints. Authors’ contribution MR, IIO, WJM and SLO had the initial idea for this manuscript. MR, IIO, WJM and SLO designed the study. MR, IIO and WJM performed laboratory work. MR, IIO, WJM, MW performed data analysis. MR, IIO, WJM, MW and SLO wrote the manuscript. All authors approved the final manuscript.

Acknowledgements We thank Sylvain Charlat, Kostas Bourtzis and the School of Veterinary Science, UQ, for supplying biological material, i.e. H. bolina, C. capitata and D. immitis, respectively. We thank the special edition editor Greg Hurst and two anonymous MLN4924 supplier reviewers for their valuable comments. Fenbendazole The research was supported by grants of the Australian Research Council ARC to MR, IIO, MW and SLO, and from COST Action FA-0701 and the research grant P22634-B17 of the Austrian Science Fund FWF to WJM. This article has been published as part of BMC Microbiology Volume 11 Supplement 1, 2012: Arthropod symbioses: from fundamental studies to pest and disease mangement. The full contents of the supplement are available online at http://​www.​biomedcentral.​com/​1471-2180/​12?​issue=​S1. References 1. Werren JH, Windsor D, Guo LR: Distribution of Wolbachia among neotropical arthropods.