The positive effects of phytases on the environment, animal welfare and animal feed costs have resulted in the continuous development and improvement of these enzymes in the non-ruminant feed market. To test the efficacy of these phytases, a large number of experimental animals are necessary, antagonising the animal welfare aspect of these enzymes. In the present review, we summarise the most prominent available in vitro assays for evaluating phytase enzymes and how far they can reduce the number of in vivo experiments. Several in vitro assays exist that differ in their setup, extent and conditions depending on the animal of interest and the research question. With the in vitro assays described, it is not possible to fully replace in vivo trials. However, for the investigation of phytase effects in feedstuffs, the use of an in vitro assay has several advantages. In vitro assays have the potential to be used for ranking feed enzymes and as screening tools. Applying in vitro protocols will result in a reduction in the number of animals or treatments usually necessary for an in vivo trial, thus acting towards the three Rs. © 2020 Society of Chemical Industry.
Keywords: in vitro; non-ruminant; phytase; phytate.
© 2020 Society of Chemical Industry.