Wolbachia is a genus of intracellular alpha-Proteobacteria that is maternally inherited and is capable of inducing a variety of reproductive alterations in host species. The host range of Wolbachia is not determined completely but is known to contain a number of arthropod taxa, including crustaceans. Wolbachia has not been reported in crayfish, but sampling has been limited to date. We examine a species of crayfish, Procambarus clarkii, for Wolbachia infection using a suite of Wolbachia-specific primers in PCR assays. All specimens yielded negative results for Wolbachia infection and mathematical analysis of sample size shows a near 100% probability of detection for populations with greater than 0.1% infection rate. 1. Introduction There is much interest among evolutionary biologists in the genus Wolbachia, a maternally inherited intracellular bacterium within the order Rickettsiales [1–3]. This interest stems from Wolbachia’s large host range [4, 5] and from its effects on host reproductive biology, including cytoplasmic incompatibility, host feminization of genetic males, induction of parthenogenesis, and male killing (reviewed by [3]). Cytoplasmic incompatibility (CI) causes incompatibility between the sperm of infected males and the eggs of uninfected females [6]. Feminization is a Wolbachia induced phenotypic change observed in both insects [7–9] and isopods (Bouchon et al. 1998, [10–12]) that results in a genetic male host developing as a reproductively functional female. Parthenogenesis induction in hosts causes the asexual production of female offspring [13–15]. Wolbachia induced male killing occurs during embryogenesis and is theorized to give a fitness advantage to surviving female offspring [16–18]. Owing to the wide range of phenotypic alterations in Wolbachia-infected hosts, there has been much recent interest in the use of Wolbachia as biological control agents [19]. The host range for Wolbachia is not determined fully, but it has been estimated that in insects alone 1.69 to 5.07 million species are infected [4]. Wolbachia is also found in filarial nematodes [20], isopods [12], and mites [21]. Intertaxon transmission is consistent with aspects of Wolbachia phylogeny [22] and has been shown to occur between Drosophila simulans and the parasitoid Leptopilina boulardi under lab conditions [23]. Wolbachia intertaxon transmission has also been implicated in Australian spiders [24] and a variety of crustacean hosts [25], suggesting that the host range of Wolbachia may be broader than is currently assumed. However, attempts to detect Wolbachia in
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