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We establish a necessary condition that an automorphism of a nontrivial finitely generated bi-orderable group can preserve a bi-ordering: at least one of its eigenvalues, suitably defined, must be real and positive. Applications are given to knot theory, spaces which fibre over the circle and to the Heegaard-Floer homology of surgery manifolds. In particular, we show that if a nontrivial fibred knot has bi-orderable knot group, then its Alexander polynomial has a positive real root. This implies that many specific knot groups are not bi-orderable. We also show that if the group of a nontrivial knot is bi-orderable, surgery on the knot cannot produce an -space, as defined by Ozsv'ath and Szab'o.

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