Organic wastes rich in phosphorus (P) are considered an alternative to decrease the dependence on chemical P fertilization in crops and pastures. Microbial inoculants are being studied as a tool to increase plant P availability in organic wastes. In this study, we explore the effect of inoculation with Bacillus sp. MQH-19 (a native phytase-producing bacterium) on the release of inorganic phosphorus (Pi) in cattle manure with low available P but a high total P content. Bacteria inoculation resulted in a higher release of Pi (8% in NaHCÜ3 and 13% in NaOH-EDTA extracts) compared with that of uninoculated manure (0.7% in NaHCÜ3 and 0.1% in NaOH-EDTA extracts). However, a greater amount of Pi was released in inoculated manure supplemented with phytate (47% in NaHCÜ3 and 117% in NaOH-EDTA extracts) compared with that of uninoculated manure supplemented with phytate (30% in NaHCÜ3 and 15% in NaOH-EDTA extracts). In addition, the use of denaturing gradient gel electrophoresis (DGGE) and quantitative PCR (qPCR) revealed that the bacterial community structure in manure was affected by inoculation and that the prevalence of Bacillus sp. MQH-19 decreased during incubation (6 days). This study demonstrates that Pi availability in cattle manure can be increased by phytase-producing bacteria inoculation. Phytase-producing bacteria inoculation might represent an attractive strategy to increase P availability in agricultural wastes, which are used as organic fertilizers in crops and pastures.