), prepatent period, site of colonization, amongst other aspects ( Coudert et al., 1995 and Pakandl, 2009). Even though oocyst morphology allows a good differentiation of rabbit Eimeria, it is practically impossible to carefully evaluate thousands of oocysts just to assess if a given strain is really pure. Moreover, a small portion of oocysts may differ from the typically expected morphology, posing a challenge Selleckchem Hydroxychloroquine to a correct diagnosis.
In addition, all these biological features may present a variable level of overlap, hampering in some cases an accurate Eimeria species identification ( Long and Joyner, 1984). In the case of chicken Eimeria, various molecular differentiation assays have been proposed, using distinct targets such as ITS1 ( Schnitzler et al., 1998 and Schnitzler et al., 1999), ITS2 ( Woods et al., 2000 and Gasser et al., 2001), and SCARs ( Fernandez et al., 2003a and Fernandez et al., 2003b). For a comprehensive review www.selleckchem.com/products/byl719.html on the molecular diagnosis of avian coccidiosis, the reader is referred to Morris and Gasser (2006). Finally, real-time PCR assays have also been described ( Blake et al., 2008 and Morgan et al., 2009), permitting quantitative studies of the distinct Eimeria species.
Conversely, no PCR-based method has been developed for the unequivocal differentiation of the 11 Eimeria species that infect rabbits. Ceré et al. (1995) described a RAPD assay for the differentiation of nine Eimeria species, but this method presents
low reproducibility. Also, the typical multiband character makes it unsuitable for testing mixed field samples. Ceré et al. (1996) developed a specific and sensitive PCR assay for E. media, but this approach has not been extended to the remaining species. The precise diagnosis of all species of rabbit coccidia is very important for several reasons. First, individual Eimeria species present dramatic differences in their pathogenicity ( Coudert et al., 1995). Second, an accurate Eimeria species differentiation method would represent an essential tool for monitoring the purity of strains in the production of a future multivalent vaccine. Finally, a correct species identification would allow research laboratories to assess however the purity of the strains under study. In this work, we report the development of molecular diagnostic assays for the detection and discrimination of the 11 Eimeria species that infect the domestic rabbit. Isolates of the 11 Eimeria species that infect the domestic rabbit (Oryctolagus cuniculus f. domesticus) were used throughout the work. E. coecicola, E. flavescens, E. irresidua, E. piriformis, E. stiedai and E. vejdovskyi isolates were originally obtained from field samples in the Czech Republic. E. exigua, E. intestinalis, E. magna, E. media and E. perforans were isolated in France.