Cytological detection of Wolbachia in squashed and paraffin embedded insect tissues

Abstract

Wolbachia localization in situ is essential for accurate analysis of the infection and its consequences. Whole cell hybridization is proposed as an easy and rapid method for detecting Wolbachia cells in paraffin embedded tissues or testis squashes of Chorthippus parallelus (Orthoptera). Wolbachia is found in whole gonads and other adjacent tissues. A higher bacterial density, however, is observed in ovarioles and testis. Small independent bacteria with strictly cytoplasmic distribution are displayed. Bacterial density differences among individuals are also revealed.

Key words::

• Chorthippus parallelus
• paraffin tissues
• whole-cell hybridization
• Wolbachia

Acknowledgments

We thank Jonás Sarasa, Maria Enciso, Mario Zabal-Aguirre, Francisca Arroyo-Yebras and the other members of our team for their contributions. This work was supported by Spanish grants CGL2005-02898/BOS and BFU2007-66340. We are especially grateful to Drs. J. MacDonald and P.L. Mason for their valuable help and advice.