The immobilisation of bacteria on biochar has shown potential for enhanced remediation of petroleum hydrocarbon-contaminated soil. However, there is a lack of knowledge regarding the effect of bacterial immobilisation on biosolids-derived biochar for the remediation of diesel-contaminated soil. This current study aimed to assess the impact of the immobilisation of an autochthonous hydrocarbonoclastic bacteria, Ochrobacterium sp. (BIB) on biosolids-derived biochar for the remediation of diesel-contaminated soil. Additionally, the effect of fertiliser application on the efficacy of the BIB treatment was investigated. Biochar (BC) application alone led to significantly higher hydrocarbon removal than the control treatment at all sampling times (4887–11,589 mg/kg higher). When Ochrobacterium sp. was immobilised on biochar (BIB), the hydrocarbon removal was greater than BC by 5533 mg/kg and 1607 mg/kg at weeks 10 and 22, respectively. However, when BIB was co-applied with fertiliser (BIBF), hydrocarbon removal was lower than BIB alone by 6987–11,767 mg/kg. Quantitative PCR (q-PCR) analysis revealed that the gene related to Ochrobacterium sp. was higher in BIB than in the BC treatment, which likely contributed to higher hydrocarbon removal in the BIB treatment. The results of the q-PCR analysis for the presence of alkB genes and FTIR analysis suggest that the degradation of alkane contributed to hydrocarbon removal. The findings of this study demonstrate that bacterial immobilisation on biosolids-derived biochar is a promising technique for the remediation of diesel-contaminated soil. Future studies should focus on optimising the immobilisation process for enhanced hydrocarbon removal.